Study On Mango Leaf and Mangiferin

Study On Mango Leaf and Mangiferin
Study on Mango Leaf and Mangiferin

Chief Editor
Deng Jiagang
Coeditor
Qin Jieping Wang Qin
Editors
Hou Xiaotao Feng Xu Liang Jianqin Hao Erwei He Cuiwei Yang Ke Wang Zhiping Zhou jiangyu Du Chengzhi Li Zhenjuan Yan Li Du Zhengcai Dai Hang Qin Lilan Shi Xueli
THE 1st INTERNATIONAL SYMPOSIUM ON SCREENING
FUNCTIONAL COMPONENTS OF AGRICULTURAL RESIDUES AND THE STUDY ON MANGIFERIN
Sponsored by:
Guangxi Traditional Chinese Medical University Management Committee of Guangxi Baise National Agricultural Sci-tech Zone
Approval Administration:
The Peoples Government of Guangxi Zhuang Autonomous Region
Supported by:
National Natural Science Foundation of China Department of Science and Technology of Guangxi Zhuang Autonomous Region
Organized by:
Faculty of Pharmacy,Guangxi Traditional Chinese Medical University Guangxi Key Laboratory of pharmacodynamic studies of Traditional Chinese Medicine
October 23-25,2009 Baise,Guangxi,Ch
Editorial Committee
Chairman
Yang Yanyang Deng Jiagang
Vice-Chairman
Zhong Hengqin Tang Qianli
Members of editorial Committee

Su Xiudong Jiang Jichang Zheng Zuowen Huang Zhaoming Nong Jinghai Lu Xiangyang Chen Yong Qin Huazhen Kuang Song Wang Qin Qin Jieping He Guibai Huang Chunxue
CONTENT
Preface ...................................................................................................................................................... 1 Invited Lecture ........................................................................................................................................ 3 The Strategic Significance and General Thoughts of the Medicinal Study On Agricultural Residues.................................................................................................................................................... 3 Chemical Constituents with Unprecedented Skeletons from Alpinia katsumadai and Chukrasia tabularis var. velutina ............................................................................................................................ 12 Study on Bioactive Compounds with Molecular Diversity from Toxic Plants in China .......................... 14 Pharmacology and Toxicology .............................................................................................................. 15 Assessment of systemic interaction between swertia chirata extract and its bioactive constituents in rabbits...................................................................................................................................................... 15 The extraction of mangiferin from mango leaves and its analgesic function............................................ 15 The Effect of Kampo Formulae on Bone Resorption in Vitro and in Vivo. I Active Constituents of Tsu-kan-gan............................................................................................................................................. 16 Mangiferin and hesperidin metabolites are absorbed from the gastrointestinal tract of pigs after oral ingestion of a Cyclopia genistoides (honeybush tea) extract .................................................................. 17 Pharmacokinetics of mangiferin in rat plasma after oral administration of a single dose of suanzaoren decoction ................................................................................................................................................. 18 Simultaneous estimation of mangiferin and four secoiridoid glycosides in rat plasma using liquid chromatography tandem mass spectrometry and its application to pharmacokinetic study of herbal preparation............................................................................................................................................... 19 UV/vis, 1H, and 13C NMR spectroscopic studies to determine mangiferin pKa values.......................... 20 A review of the bioactivity of south african herbal Teas: rooibos (aspalathus linearis) and honeybush (Cyclopia intermedia) ............................................................................................................................. 21 Activation of lymphocytes of normal and tumor bearing mice by mangiferin, a naturally occurring glucosylxanthone..................................................................................................................................... 21 An Anacardiaceae preparation reduces the expression of inflammation-related genes in murine macrophages............................................................................................................................................ 22 Anthelminthic and antiallergic activities of Mangifera indica L. Stem bark components vimang and mangiferin ............................................................................................................................................... 23 Anti-allergic properties of Mangifera indica L. extract (Vimang) and contribution of its glucosylxanthone mangiferin .................................................................................................................. 24 Antidiabetic activity of a xanthone compound, mangiferin ...................................................................... 25 Antidiabetic activity of the rhizoma of anemarrhena asphodeloides and active components, mangiferin and its glucoside ...................................................................................................................................... 25

Antiinflammatory, analgesic and hypoglycemic effects of Mangifera indica Linn. (Anacardiaceae) stem-bark aqueous extract....................................................................................................................... 26 Antitumor, immunomodulatory and anti-HIV effect of mangiferin, a naturally occurring glucosylxanthone..................................................................................................................................... 27 Chemopreventive efficacy of mangiferin against benzo(a)pyrene induced lung carcinogenesis in experimental animals............................................................................................................................... 27 -D-Glucoside suppresses tumor necrosis factor-induced activation of uclear transcription factor B but potentiates apoptosis ............................................................................................................................... 28 Cytoprotective and antigenotoxic potential of Mangiferin, a glucosylxanthone against cadmium chloride induced toxicity in HepG2 cells .............................................................................................................. 29 Cytoprotective effect of mangiferin on benzo(a) pyrene-induced lung carcinogenesis in swiss albino mice ......................................................................................................................................................... 30 Differential oxidative stress in oligodendrocytes and neurons after excitotoxic insults and protection by natural polyphenols ................................................................................................................................. 31 Dual mechanism of mangiferin protection against iron-induced damage to 2-deoxyribose and ascorbate oxidation.................................................................................................................................................. 32 Effect of Mangifera indica L. extract (QF808) on protein and hepatic microsome peroxidation ............. 33 Effect of mangiferin on benzo(a)pyrene induced lung carcinogenesis in experimental Swiss albino mice34 Efficacy of mangiferin on serum and heart tissue lipids in rats subjected to isoproterenol induced cardiotoxicity........................................................................................................................................... 34 Effect of mangiferin on hyperglycemia and atherogenicity in streptozotocin diabetic rats ...................... 35 Effect of mangiferin on mitochondrial energy production in experimentally induced myocardial infarcted rats............................................................................................................................................ 36 Effect of mangiferin on radiation-induced micronucleus formation in cultured human peripheral blood lymphocytes ............................................................................................................................................ 37 Effect of mangiferin on the development of periodontal disease: involvement of lipoxin A4, anti-chemotaxic action in leukocyte rolling ............................................................................................ 38 Effect of species variation and processing on phenolic composition and in vitro antioxidant activity of aqueous extracts of Cyclopia spp. (Honeybush Tea) .............................................................................. 39 Effects of a natural extract from Mangifera indica L, and its active compound, mangiferin, on energy state and lipid peroxidation of red blood cells......................................................................................... 40 Effects of the mango components mangiferin and quercetin and the putative mangiferin metabolite norathyriol on the transactivation of peroxisome proliferator-activated receptor isoforms .................... 41 Efficacy of mangiferin against Cryptosporidium parvum in a neonatal mouse model ............................. 42 Evaluation of the genotoxic potential of Mangifera indica L. extract (Vimang), a new natural product with antioxidant activity.......................................................................................................................... 43

Examination of the inhibitory effect of norathyriol in formylmethionyl-leucyl-phenylalanine-induced respiratory burst in rat neutrophils .......................................................................................................... 44 Expression profiles of genes involved in the mouse nuclear factor-kappa B signal transduction pathway are modulated by mangiferin................................................................................................................... 45 Fe (III) improves antioxidant and cytoprotecting activities of mangiferin................................................ 46 Immunomodulatory activity of alcoholic extract of Mangifera indica L. in mice .................................... 46 Gastroprotective effect of mangiferin, a xanthonoid from Mangifera indica, against gastric injury induced by ethanol and indomethacin in rodents .................................................................................... 47 In vitro effects of Mangifera indica and polyphenols derived on ABCB1/P-glycoprotein activity .......... 48 Immunotherapeutic effects of mangiferin mediated by the inhibition of oxidative stress to activated lymphocytes, neutrophils and macrophages............................................................................................ 49 In vitro effects of mangiferin on superoxide concentrations and expression of the inducible nitric oxide synthase, tumournecrosis factor- and transforming growth factor- genes........................................... 50 In vitroeffects of the polyphenols resveratrol, mangiferin and (-)-epigallocatechin-3-gallate on the scuticociliate fish pathogen Philasterides dicentrarchi............................................................................ 51 Mangiferin,a glucosylxanthone,protects against the radiation-induced micronuclei formation in the cultured human peripheral blood lymphocytes ....................................................................................... 51 In vivo and in vitro anti-inflammatory activity of Mangifera indica L. extract (VIMANG).................... 52 Insulin secretion is stimulated by ethanol extract of anemarrhena asphodeloides in isolated islet of healthy wistar and diabetic Goto-Kakizaki Rats ..................................................................................... 53 Interaction of Vimang (Mangifera indica L. extract) with Fe(III) improves its antioxidant and cytoprotecting activity............................................................................................................................. 54 Iron complexing activity of mangiferin,a naturally occurring glucosylxanthone,inhibits mitochondrial lipid peroxidation induced by Fe2+-citrate............................................................................................... 55 Isolation of a human intestinal bacterium that transforms mangiferin to norathyriol and inducibility of the enzyme that cleaves a C-Glucosyl bond............................................................................................ 56 Mangifera indica L. extract (Vimang) and its main polyphenol mangiferin prevent mitochondrial oxidative stress in atherosclerosis-prone hypercholesterolemic mouse. ................................................. 57 Mangifera indica L. extract (Vimang) and mangiferin modulate mouse humoral immune responses. .... 58 Mangifera indica L. extract (Vimang) inhibits 2-deoxyribose damage induced by Fe (III) plus ascorbate.59 Mangifera indica L. extract (Vimang) inhibits Fe2+-citrate-induced lipoperoxidation in isolated rat liver mitochondria............................................................................................................................................ 60 Mangifera indica L. extract attenuates glutamate-induced neurotoxicity on rat cortical neurons. ........... 61 Mangiferin ameliorates scopolamine-induced learning deficits in mice ................................................... 62 Mangiferin inhibits cyclooxygenase-2 expression and prostaglandin E2 production in activated rat glial

cells ......................................................................................................................................................... 63 Mangiferin Inhibits Passive Cutaneous Anaphylaxis Reaction and Pruritus in Mice ............................... 64 Mangiferin protects against 1-methyl-4-phenylpyridinium toxicity mediated by oxidative stress in N2a cells ......................................................................................................................................................... 65 Mangiferin protects human peripheral blood lymphocytes against -radiation-induced DNA strand breaks: a fluorescence analysis of DNA unwinding assay ...................................................................... 66 Mangiferin protects the streptozotocin-induced oxidative damage to cardiac and renal tissues in rats .... 67 Mangiferin, a natural occurring glucosyl xanthone, increases susceptibility of rat liver mitochondria to calcium-induced permeability transition ................................................................................................. 68 Mangiferin protects human peripheral blood lymphocytes against -radiationinduced DNA strand breaks:a fluorescence analysis of DNA unwinding assay ....................................................................... 69 Mechanism of Antioxidant Action of Pueraria Glycoside (PG)-1 (an Isoflavonoid) and Mangiferin (a Xanthonoid)............................................................................................................................................. 70 Mechanism of cell death induced by an antioxidant extract of Cratoxylum cochinchinense (YCT) in Jurkat T cells: the role of reactive oxygen species and calcium.............................................................. 71 Mechanism of protective action of mangiferin on suppression of inflammatory response and lysosomal instability in rat model of myocardial infarction ..................................................................................... 71 Mechanisms of blood glucose-lowering effect of aqueous extract from stems of Kothala himbutu (Salacia reticulata) in the mouse ............................................................................................................. 72 Modulation of P450 enzymes by Cuban natural products rich in polyphenolic compounds in rat hepatocytes .............................................................................................................................................. 73 Modulation of rat macrophage function by the Mangifera indica L. extracts Vimang and mangiferin .... 74 Molecular mechanisms of neuroprotection by two natural antioxidant polyphenols................................ 75 New antidiabetic compounds, mangiferin and its glucoside ..................................................................... 75 New antioxidant C-glucosylxanthones from the stems of Arrabidaea samydoides .................................. 76 Neuroprotection by two polyphenols following excitotoxicity and experimental ischemia ..................... 76 Novel screening assay for antioxidant protection against peroxyl radical-induced loss of protein function ................................................................................................................................................... 77 Pharmacokinetic study of free mangiferin in rats by microdialysis coupled with microbore high-performance liquid chromatography and tandem mass spectrometry............................................. 78 Physiological and biochemical changes with special reference to mangiferin and oxidative enzymes level in malformation resistant and susceptible cultivars of mango (Mangifera indica L.) .................... 79 Polyphenols with antiulcerogenic action from aqueous decoction of mango leaves (Mangifera indica L.)80 Potential hepatoprotective effects of new Cuban natural products in rat hepatocytes culture .................. 81 Protection against septic shock and suppression of tumor necrosis factor and nitric oxide production

on macrophages and glia by a standard aqueous extract of Mangifera indica L. (VIMANG). Role of mangiferin isolated from the extract ....................................................................................................... 82 Protective effect of Mangifera indica L. polyphenols on human T lymphocytes against activation-induced cell death................................................................................................................... 83 Protective effects of a standard extract of Mangifera indica L. (VIMANG) against mouse ear edemas and its inhibition of eicosanoid production in J774 murine macrophages .............................................. 84 Protective effects of Mangifera indica L. extract (Vimang), and its major component mangiferin, on iron-induced oxidative damage to rat serum and liver ............................................................................ 85 Protective effects of Mangifera indica L. extract, mangiferin and selected antioxidants against TPA-induced biomolecules oxidation and peritoneal macrophage activation in mice ........................... 86 Protective role of mangiferin against Benzo(a)pyrene induced lung carcinogenesis in experimental animals .................................................................................................................................................... 87 Studies on palauan medicinal herbs. II. Activation of mouse macrophages RAW 264.7 by Ongael, leaves of Phaleria cumingii (Meisn.) F. Vill. and its acylglucosylsterols ............................................... 87 Radioprotection by mangiferin in DBAxC57BL mice: a preliminary study .............................................. 88 The suppressive effect of mangiferin with exercise on blood lipids in type 2 diabetes ............................ 88 Release of intermediate reactive hydrogen peroxide by macrophage cells activated by natural products 89 Role of mangiferin on biochemical alterations and antioxidant status in isoproterenol-induced myocardial infarction in rats.................................................................................................................... 90 Salacia oblonga extract increases glucose transporter 4-mediated glucose uptake in L6 rat myotubes: Role of mangiferin .................................................................................................................................. 91 Salacia oblonga improves cardiac fibrosis and inhibits postprandial hyperglycemia in obese Zucker rats92 Salacia oblonga root decreases cardiac hypertrophy in Zucker diabetic fatty rats: inhibition of cardiac expression of angiotensin II type 1 receptor............................................................................................ 93 Salacia reticulata and its polyphenolic constituents with lipase inhibitory and lipolytic activities have mild antiobesity effects in rats................................................................................................................. 94 Scavenger effect of a mango (Mangifera indica L.) food supplement's active ingredient on free radicals produced by human polymorphonuclear cells and hypoxanthine-xanthine oxidase chemiluminescence systems .................................................................................................................................................... 95 Spectroscopic investigation of interaction between mangiferin and bovine serum albumin .................... 95 Swertia chirayita mediated modulation of interleukin-1beta, interleukin-6, interleukin-10, interferon-, and tumor necrosis factor- in arthritic mice .......................................................................................... 96 The inhibitory effects of mangiferin, a naturally occurring glucosylxanthone, in bowel carcinogenesis of male F344 rats ......................................................................................................................................... 97 The variation in cytoplasmic distribution of mouse peritoneal macrophage during phagocytosis modulated by mangiferin, an immunomodulator .................................................................................... 98

Timosaponin AIII, a saponin isolated from Anemarrhena asphodeloides, ameliorates learning and memory deficits in mice .......................................................................................................................... 99 Two proteins, Mn2+, and low molecular cofactor are required for C-glucosyl-cleavage of mangiferin . 100 Utilization of mango peels as a source of pectin and polyphenolics....................................................... 101 Vascular effects of the Mangifera indica L. extract (Vimang) ............................................................... 102 Vimang (Mangifera indica L. extract) induces permeability transition in isolated mitochondria, closely reproducing the effect of mangiferin, Vimang's main component ........................................................ 103 Xanthone derivatives: new insights in biological activities .................................................................... 104 Xanthone glycosides from herbs of Polygala hongkongensis Hemsl and their antioxidant activities .... 104 Pharmacokinetics of mangiferin in rat plasma after oral administration of a single dose of Suanzaoren decoction ............................................................................................................................................... 105 Synthesis of mangiferin derivates and study their potent PTP1B inhibitory activity.............................. 105 Pharmacokinetics of mangiferin in rat plasma after oral administration of a single dose of Suanzaoren decoction ............................................................................................................................................... 106 Effect of Mangifer in on telomerase activity and cell cycle in K562 cells.............................................. 107 Effect of Mangiferin on the Content of PGE2 in Two Different Inflammation Models.......................... 107 The Effect of Mangifer in on Telomerase Activity and Apoptosis in Leukem ic K562 Cells ................ 108 The Antitussive and Expectorant Effects of Mangifera Leaves Extract.................................................. 108 Preliminary Studies on the Mode of Action of Mangifer in against Phytophthora infestans.................. 109 Experimental Study on Anti-bacterial, Anti-inflammatory and Analgesic Activities of the Mixture of Mangiferin and Berberine ..................................................................................................................... 109 An Experimental Study of Anti-stress Effects of Mangiferin in Mice.................................................... 110 Preliminary study on effects of mangiferin on immunologic function in mice....................................... 110 Effects of mangiferin of TNF- and MPO in rats with myocardial ischemia reperfusion injury............ 111 Experimental Study on Hypoglycemic Effect of the Mixture of Mangiferin and Berberine .................. 111 Effect of mangiferin on myocardial ischemia induced by isoproteronol in mice.................................... 112 Apoptotic mechanism of leukemic K562 cells induced by mangiferin................................................... 112 Inhibitory effect of mangiferin on duck hepatitisB virus (DHBV) DNA in vivo.................................... 113 Comparison Tests of the Efficacy of Mango Leaf Decoction, Demangiferin Mango Leaf Decoction and Mangiferin Anti-tussive and Expectorant Drugs................................................................................... 113 Study on Antibacterial Action of Extract of Leaves of Mangifera indica in Vitro ................................. 114 Inhibiting effect in vitro of extract of Mangifera indica L.leaf on some pathogenic bacteria and NDV replication.............................................................................................................................................. 114

The Ultramicro-structure Change of Lipid Superoxided Rat Brain Tissue and Protect of Mangiferin on the Tissues............................................................................................................................................. 115 Inhibitory effect of mangiferin on the proliferation of K562 leukemia cells .......................................... 116 Effect of mangiferin on the expression of -cateninand p120ctn in hepatic tissues of rats with liver cancer .................................................................................................................................................... 117 Effects of enzyme and morphological change of mangiferin on experimental liver damage in rats....... 117 The proliferation inhibition effect and apoptosis induction of mangiferin on BEL-7404 human hepatocellular carcinoma cell................................................................................................................ 118 Effect of mangiferin on P120ctn phosphorylation and hepatocellular carcinoma cell biology............... 118 Experimental study on effect of mangiferin delaying caducity............................................................... 119 Effects of Mangifer in on induction of apoptosis and in tracellular Ca2+ concentration in Nasopharyngeal Carcinoma CNE2 Cells .............................................................................................. 119 CML cell line K562 cell apoptosis induced by Mangiferin .................................................................... 120 Pharmacodynamic studie s on Mangiferin .............................................................................................. 120 Experimental study on the pharmacology of Mangiferin monosodium salt............................................ 121 Preparation of mengiferin monosodium salt and comparison in pharmacological effects with mengiferin121 Protective effect of Mangiferin dropping pills on chronic liver injury in rats......................................... 122 Effects of Mangiferin on cell cycle status and cyclin A,cyclin B1 expression of K562 cells ................. 123 Effects of Mangiferin on myocardial ischemia induced by pituitrin in mice .......................................... 123 Effects of Mangiferin on Gastric Ulcers in Rats ..................................................................................... 124 Effect of Mangiferin on the arachidonic acid metabolites in rat ............................................................. 124 Cardioprotective Effects of Mangiferin on Myocardial in Schemia Reperfusion Injury in Rats ............ 125 Effect of Mangiferin on lymphocyte proliferation in immunosuppressed mice...................................... 125 The effects of Mangiferin on human platelet aggregation and secretion of CD62P ............................... 126 Effect of Mangifer on Serum E-cadherin, carcinoembryonic antigen and monoamine oxidase activity and cell cycle in live tumor rats............................................................................................................. 127 The Impact of Mangiferin on Releasing of Slow Reacting Substance of Anaphylaxis from Guinea-Pig Lung Tissue ........................................................................................................................................... 127 Effects of on Mangiferin HBsAg and HBeAg Excreted by 2215 Cell.................................................... 128 The Effect of Mangiferin on hTERT-mRNA Expression and Telomerase Activity in K562 Cells........ 128 Protective effect of Mangiferin on Alcohol-Induced Liver Injury in Mice............................................. 129 Pharmacodynamic Study of Total Glycosides Tablet of Mango Leaves ................................................ 129 The influence of mangiferin on the body temperature of rabbit in endotoxin-induced fever ................. 130

The study on mangiferin protective role of lipid peroxidation damage of brain tissues in rats .............. 130 The antidepressant effect of mangiferin on the behavioral despair mice ................................................ 131 Effects of Mangiferin on lipid peroxidation metabolism of blood in rats ............................................... 131 Antlviral activity of mangiferin against herpes simplex virus type 2 in vitro ......................................... 132 Effect of Mangiferin on the Arachidonic Acid Metabolizing Enzymes in Rat Neutrophils ................... 132 Antiviral effect of mangiferin and Isomangiferin on herpes simplex virus................................. 133
Chemical Study and Analytical methods .......................................................................................... 134 Characterization and quantitative determination of the impurity in prepared mangiferin extracted from Mangifera indica L. leaves.................................................................................................................... 134 A new C-glycosyl xanthone isolated from Davallia solida .................................................................... 151 Characterization and quantitation of polyphenolic compounds in bark, kernel, leaves, and peel of mango (Mangifera indica L.) ................................................................................................................ 151 An investigation of the stem bark of Bersama abyssinica ...................................................................... 152 Biosynthesis of mangiferin in anemarrhena asphodewides: intact incorporation of C6-C3 precursor into xanthone ................................................................................................................................................ 152 Antiosteoporotic chemical constituents from Er-Xian Decoction,a traditional Chinese herbal formula 153 Antioxidant C-Glucosylxanthones from the Leaves of Arrabidaea patellifer ........................................ 154 A Xanthone C-glycoside from Iris Nigricans ......................................................................................... 154 Benzophenone glycosides from Gnidia involucrate ............................................................................... 155 Capillary electrophoresis analysis of mangiferin extracted from Mangifera indica L. bark and Mangifera persiciformis C.Y. Wu et T.L. Ming leaves .......................................................................................... 155 Characterizaton of antioxidant and antiglycation properties and isolation of active ingredients from traditional Chinese medicines ............................................................................................................... 156 Chemical constituents from Mahkota dewa ............................................................................................ 156 Characterization of polyphenols in mango puree concentrate by HPLC with diode array and mass spectrometric detection ......................................................................................................................... 157 Characterization of the mangiferin-human serum albumin complex by spectroscopic and molecular modeling approaches............................................................................................................................. 157 Chemical and chemotaxonomical studies of ferns. LXXXVII. constiuents of trichomanes reniforme .. 158 Chemical constituents of Gentianaceae XIX: CNS-depressant effects of swertiamarin ......................... 158 Determination of gentiopicroside, mangiferin, palmatine, berberine, baicalin, wogonin and glycyrrhizin in the traditional Chinese medicinal preparation Sann-Joong-Kuey-Jian-Tang by high- performance liquid chromatography .......................................................................................................................... 159 Determination of the residue of organochlorine pesticides in mango leaves using GC-MS-SIM........... 159

Determination of mangiferin, jateorrhizine, palmatine, berberine, cinnamic Acid, and cinnamaldehyde in the traditional Chinese medicinal preparation Zi-Shen Pill by high-performance liquid chromatography..................................................................................................................................... 160 Differentiation of Swertia Mussotii Franch from Artemisiae Capillaris Herba by capillary electrophoresis with electrochemical detection..................................................................................... 160 Evaluation of spectrophotometric methods for screening of green rooibos (Aspalathus linearis) and green honeybush (Cyclopia genistoides) extracts for high levels of Bio-active compounds ................ 161 Flavonoid and xanthone patterns in bearded Iris species and the pathway of chemical evolution in the genus ..................................................................................................................................................... 162 Glucuronide triterpene saponins from Bersama engleriana.................................................................... 163 High-performance liquid chromatographic method for the determination and pharmacokinetic study of mangiferin in plasma of rats having taken the traditional Chinese medicinal preparation Zi-Shen pill 164 High-performance liquid chromatography as a tool for the chemical standardisation of triphala-an ayurvedic formulation ........................................................................................................................... 164 Isolation of isomangiferin from honeybush (Cyclopia subternata) using high-speed counter-current chromatography and high performance liquid chromatography ........................................................... 165 Isolation of Mangiferin and Isomangiferin from Leaf Material of Hibiscus liliastrum (Malvaceae) ..... 166 Isolation of mangiferin from Bombax malabaricum and structure revision of shamimin....................... 166 Mangiferin and isomangiferin in some Hypericum species .................................................................... 167 Mangiferin Identified in a Screening Study Guided by Neuraminidase Inhibitory Activity................... 167 Liquid chromatography/tandem mass spectrometric study and analysis of xanthone and secoiridoid glycoside composition of Swertia chirata, a potent antidiabetic ........................................................... 168 Mangeiferin from the root bark of salaczaretzculata............................................................................... 169 Miscibility Characterization in Relation to Phase Morphology of Poly (ether sulfone)/Poly (vinyl pyrrolidone) Blends Containing a Phytochemical................................................................................. 170 New Steroidal Sapomins from the Rhizomes of Anemarrhena asphodeoides Bunge (Liliaceae) .......... 170 On-line purity monitoring in high-speed counter-current chromatography Application of HSCCC-HPLC-DAD for the preparation of 5-HMF, neomangiferin and mangiferin from Anemarrhena asphodeloides Bunge............................................................................................................................. 171 Phenolic compounds from Hypericum perforatum ................................................................................. 172 Phenolic metabolites from honeybush tea (cyclopia subternata) ............................................................ 172 Polyphenol constituents from salacia species: quantitative analysis of mangiferin with a-glucosidase and aldose reductase inhibitory activities .................................................................................................... 173 Preparative isolation and purication of four compounds from the chinese medicinal herb rhizoma anemarrhenae by high-speed counter-current chromatography ............................................................ 174

Chemical constituents in the leaves of Mangifera persiciformis C.Y. Wu et Y.L. Ming ....................... 174 Quality evaluation of rhizoma belamcandae (belamcanda chinensis (L.) DC.) by using high-performance liquid chromatography coupled with diode array detector and mass spectrometry . 175 Synthesis of mangiferin........................................................................................................................... 175 Rapid Identification of Polyphenol C-Glycosides from Swertia franchetiana by HPLC-ESI-MS-MS .. 176 Secoiridoids and Xanthones from Gentianella nitida .............................................................................. 176 Simultaneous determination of bioactive xanthone glycosides and norlignans from ethanolic extract of Anemarrhena asphodeloides by liquid chromatography....................................................................... 177 Simultaneous determination of phenols in Radix Polygalae by high performance liquid chromatography: quality assurance of herbs from different regions and seasons ............................................................. 178 Simultaneous estimation of mangiferin and four secoiridoid glycosides in rat plasma using liquid chromatography tandem mass spectrometry and its application to pharmacokinetic study of herbal preparation............................................................................................................................................. 179 Structures of New Friedelane-Type Triterpenes and Eudesmane-Type Sesquiterpene and Aldose Reductase Inhibitors from Salacia chinensis......................................................................................... 180 Studies on the constituents from the fruits of Phaleria macrocarpa ........................................................ 180 Synthesis of mangiferin derivates and study their potent PTP1B inhibitory activity.............................. 181 The Major Phenolic Compounds in the Leaves of Cyclopia Species (Honeybush Tea)......................... 181 Temperature and solvent dependent NMR studies on mangiferin and complete NMR spectral assignments of its acyl and methyl derivatives ..................................................................................... 182 Use of NIRS for quantification of mangiferin and hesperidin contents of dried green honey bush (Cyclopia genistoides) plant material................................................................................................. 182 Using LC/MS/MS to determine matrine, oxymatrine, ferulic acid, mangiferin, and glycyrrhizin in the Chinese medicinal preparations Shiau-feng-saan and Dang-guei-nian-tong-tang ................................ 183 Variation of active constituents of an important Tibet folk medicine Swertia mussotii Franch. (Gentianaceae) between artificially Cultivated and naturally distributed............................................. 183 UV/vis, 1H and 13CNMR spectroscopic studies to determine mangiferin pKa values ........................... 184 Quality standard research on Mangiferin crude drug .............................................................................. 184 Xanthones from Swertia punctata ........................................................................................................... 185 Study on the extracting method of mangiferin in Anemarrhena asphodeloides Bge and Comparison of content of mangiferin in hair, skin and meat of Anemarrhena asphodeloides Bge............................... 185 Study of extraction and stability of yellow pigment from mango leaves ................................................ 186 Extraction and identification of total flavone from mango leaves .......................................................... 186 Extraction and identification of mangiferin from Mangifera indica leaves ............................................ 187 Extraction of total flavanone from mango leaves by ultrasonic wave..................................................... 187
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Optimized procedures for quercetin extraction from Mangifera indica Linn. leaf based on orthogonal design .................................................................................................................................................... 188 Determination of mangiferin and homomangiferin in almond leaves by HPLC..................................... 188 Determination of gallic acid in mango leaves by HPLC ......................................................................... 189 Study on the content of mangiferin in mangifera indica L. from different areas.................................... 189 Determination of mangiferin and homomangiferin in manggo leaves by HPLC.................................... 190 Determination of impurity of homomangiferin in raw medicine of mangiferin by HPLC ..................... 190 Comparison of HPLC fingerprint among different tissues of Mangifera indica L. ................................ 191 Comparison research on the content of mangiferin between manggo leaf and manggo branch ............. 191 Determination of gallic acid in the leaves of 4 genera of Mangifera indica L. by RP-HPLC ................ 192 Determination of mangiferin, neomangiferin in Rhizoma anemarrhenae from different producing area192 Comparison of mangiferin content in different cultivars of mango leaves ............................................. 193 Determination of mangiferin of the aerial parts in Gentiana manshurica Kitagawa .............................. 193 Determination of mangiferin in dejecta of rabbit by RP-HPLC.............................................................. 194 Determination of mangiferin in Qingqiliangying injection by RP-HPLC............................................... 194 Determination of the contents of mangiferin and berberine hydrochloride in the Zishen Pills by RP-HPLC .............................................................................................................................................. 195 Determination of mangiferin in rhizoma anemarrhenae from different habitats by HPLC-UV............. 195 Determination of mangiferin in rhizoma anemarrhenae from different habitats by HPLC.................... 196 Determination of mangiferin and sarsasapogenin in rhizoma anemarrhenae and stir-baked rhizoma anemarrhenae before sprinking salt solution by HPLC........................................................................ 196 Determination of mangiferin and neomangiferin in rhizoma anemarrhenae and its preparation by HPLC197 Determination of mangiferin and neomangiferin in rhizoma anemarrhenae by HPLC.......................... 197 Determination of chimonin and forsythiaside in Kangbingdu oral liquid by RP-HPLC......................... 198 Determination of Mangiferin in Mango peel by RP-HPLC .................................................................... 198
IN ORDER TO ESTABLISH A METHOD FOR THE DETERMINATION OF MANGIFERIN IN MANGO PEEL, A HPLC
METHOD WAS ESTABLISHED, AND A HANBON LICHYOPHER C18 (4.6250MM,5M) COLUMN WAS USED. THE MOBILE PHASE WAS METHANOL-0.3%H3PO4 (32:68) AND THE FLOW RATE WAS1ML/MIN. THE UV DETECTION
THE AVERAGE RECOVERY OF MANGIFERIN WAS 97.8WITH RSD OF 1.85. THIS METHOD IS SIMPLE ACCURATE REPRODUCIBLE. IT WAS FOUND THAT THE MANGO PEEL IN
EQUATION OF Y=1.065+2.035X, R=0.9999 (N=5)
WAVELENGTH WAS 258NM ,THE LINEAR RANGES OF MANGIFERIN WERE IN THE RANGE OF 0.4~0.8UG WITH
BAI-SE, NANNING, AND TIAN-YANG COUNTY HAVE THE HIGHEST CONTENT OF MANGIFERIN........................... 198
RP-HPLC determination of mangiferin in the leafs of Folium Mangiferae sampled in different months and regions ............................................................................................................................................ 199
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Determination of the contents of mangiferin in the roots and stems of Rhizoma Anemarrhenal at different harvest dates ........................................................................................................................... 199 Determination of mangiferin and neomangiferin in Rhizoma Anemarhenal using RP-HPLC................ 200 Determination of Mangiferin and Polysaccharide in Rhizoma Anemarrhenal from Different Origin .... 200 Quantitative determination of four effective components in Swertia delavayi by HPLC ....................... 201 Determination of mangiferin in Liyan tablets by HPLC ......................................................................... 201 Determination of mangiferin in Zhibai Dihuang Pill by HPLC .............................................................. 202 Dertermination content of mangiferin and sarsasapogenine in Rhizoma Anemarrhenae from different areas....................................................................................................................................................... 202 Separation of mangiferin by high performance capillary electrophoresis............................................... 203 HPLC determination of mangiferin content in commercial Rhizoma Anemarrhenae............................. 203 Comparison of Mangifern in Contents in Different Parts of Mango Tree .............................................. 204 The physiological and biochemical change induced by mangiferin accumulation in Mango tree.......... 204 Optimization of Extraction Technology for Total Saponin in Mango Leaf ............................................ 205 Rapid determination of mangiferin and neomangiferin in Anemarrhena asphodeloides Bge. By capillary zone eectrophoresis with UV detection................................................................................................. 205 Isolation and Identification of Oleanolic acid and Magiferin from Swertia punicea hemsl .................... 206 Changes of mangiferin and neomangiferin contents in Rhizoma Anemarrhenae before and after processing.............................................................................................................................................. 206 Dynamic Study of Contents of mangiferin in XiLing Anemarrhena asphodeloides Bge. ...................... 207 Study on the Factors Affecting the Mangiferin Contents in Mango Leaves ........................................... 207 Structure modification of mangiferin ...................................................................................................... 208 Identification and determination of four metabolites of mangiferin in rat urine ..................................... 208 Stability of mangiferin and factors affecting the stability ....................................................................... 209 Isolation and structure modification of mangiferin from Anemarrhena asphodeloides Bge. ................. 209 Determination of mangiferin in Zhimu Compounding Granules by HPLC ............................................ 210 High-performance liquid chromatographic method for the determination of mangiferin, ikviritin and dihydroquercetin in rat plasma and urine .............................................................................................. 210 Pharmacokinetic study of free mangiferin in rats by microdialysis coupled with microbore high-performance liquid chromatography and tandem masss pectrometry........................................... 211 High-performance liquid chromatographic method for the determination of mangiferin in rat plasma and urine....................................................................................................................................................... 212 Identification of major xanthones and steroidal saponins in rat urine by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry technology following
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oral administration of Rhizoma Anemarrhenae decoction .................................................................... 213 Purification and analysis by high performance liquid chromatography of Mangiferin........................... 214 Pulse radiolysis studies of mangiferin: A C-glycosyl xanthone isolated from Mangifera indica ........... 214 Technology and Application ............................................................................................................... 215 Summarize of mangiferin products ......................................................................................................... 215 30 Cases of acute upper Respiratory infection treated with mangiferin tablets ...................................... 218 Optimization of mangiferin extraction process by orthogonal design from Zhimu ................................ 218 Preparation of mangiferin monosodium salt ........................................................................................... 219 Solubility enhancement of mangiferin by HP--CD inclusion technic ................................................... 219 Studies on the effects of mango leaf electuary upon influenza ............................................................... 220 Study on mangiferin extraction by air-blasting method .......................................................................... 220 Study of extraction of Mango Leaf total glucosides tablets .................................................................... 221 Study on processing technology of mangiferin pills ............................................................................... 221 Study on thin-film coating process for mangiferin tablet........................................................................ 222 Study on ultrasonic extraction technics of mangiferin in Anemarrhena asphodeloides Bge.................. 222 Index ....................................................................................................................................................... 223
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Preface
The civilization and evolution of society, if getting more progress, always depend on the invention or discovery of new substance, new ways, new knowledge and new theories. We,as a positive enterprising and vigorous team devoting to medicine research, have been engaged in the study of mango leaves and mangiferin as the scholars from different countries do. We do hope to discover some new medical values from mango leaves and mangiferin so as to contribute to humans health. We are dramatically inspired, after long-term endeavors, by many encouraging reports released from all over the world. For the convenience of communication and discussion, we have collected all these achievements to form the corpus of the study of mango leaves and mangiferin. The corpus is divided into six parts preface,invited lecture,pharmacodynamics and toxicology, chemical study and analytical methods,technology and application, index.Total 291 papers are collected both including submitted papers from pharmacodynamic actions study of castoff crops and the first international symposium on mangiferin and published reports in all kinds of magazines. 337 researchers and experts of the research papers are from 29 countries; the contents of the papers involve the extracting technology, separation, purification, the chemical structure modification, toxicity, pharmacodynamic action, mechanism, and clinical research on Mango leaves and Mangiferin. The latest research report is the toxicity study of mangiferin made by professor Deng Jiagang and his partner from Guangxi, P.R.China. Generally speaking, the corpus is a great summary of long-period study, which truly reflects present research process on mangiferin. The major purpose we edit the corpus is to provide a platform for the experts and researchers to discuss and exchange their achievements in the study on mangiferin. Thanks to both the chief editors high attention to this fields and all editors hard work, the corpus could be published successfully within two months. Whats more, various circles of society also provide huge supports and financial aids for academic conference. We are sincerely grateful to Guangxi Traditional Chinese Medical Univerisity, Baise National Agricultural SCI-TECH Zone, Guangxi Science and Technology Department and National Natural Science Foundation of China (NSFC). Though we require every editor to compile the corpus as perfectly and quickly as
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possible, it is also inevitable to appear some mistakes or deficiencies due to limited time, our professional knowledge and limited English levels. Please instantly contact us if you find any mistakes or your research papers are not published in the corpus. We are sincerely thankful to those who give valuable advice on the corpus. As an ancient Chinese poet said, Although the way stretched endless ahead, we will search with my will unbending. When the conference draws to a close, when honorable guests leave the beautiful Zhuang Autonomous region, the cooperation between us is just beginning. We will firm our study direction to further research on mango leaves and mangiferin, especially their application research. We sincerely hope this piece of green leaf carrying our aborative contribution can bring the health and happiness to human.
The Organizing Committee October 2009
Invited Lecture
The Strategic Significance and General Thoughts of the Medicinal Study On Agricultural Residues
Jiagang Deng
Being engaged in the research of Chinese herbs for many years and based on the thoughts of sustainable development of the natural resources of Chinese herbs over a long period of time, the author, since the end of March 2009, in many public occasions delivers the three no-mainstream strategic considerations about the problem of sustainable development of the natural resources of Chinese herbs, which means that we should study chemical substances as new resources of Chinese herbs, carry on the medicinal study of agricultural residues and restructure the internationalization ideas of Chinese medicine and restrict the non-medical consume of Chinese herbs so as to realize sustainable development of the natural resources of Chinese herbs. In this paper, the author from the macro level tries to discuss the reasons and approaches to carry on the medicinal study on agricultural residues 1. The strategic significance of the medicinal study on agricultural residues The so-called agricultural residues are not the parts of crops for economic object planted by farmers including growers of vegetable, fruit, flower, herbs, etc., which are not agricultural products and not be used for medicinal purpose or other valuable commercial action in traditional production business. For example, vegetable growers plant tomatoes, and the fruit of tomatoes is their main economic object, and the stem and leaves of tomatoes are not their main economic object. The fruit grower plants mangoes, melons and bananas, and the fruit of mangoes, melons and bananas are their economic objects, and the mango leaves pruned among the growing and the stem and leaves after the fruit of melon and banana are picked, are not their main economic object. We call such kinds of materials of minor economic object as agricultural residues. In past production activity, most of the agricultural residues are discarded because they are not the main economic objects. What we will do is to apply modern science and technology to research the medicinal value of the agricultural residues to find their new values. We can understand the strategic significance of this research from its impacts on medicine, ecology and society.

(1) The medicinal study on the agricultural residues is an important way to support sustainable development of the natural resources of Chinese herbs. The natural resources of Chinese herbs are the basis of Chinese medicine (Traditional Chinese Medicine). The existence and development of Chinese medicine depends largely on the sustainable development of the natural resources of Chinese herbs. Development and utilization of the natural resources of Chinese herbs enjoys a long history in our country, and there are 12,772 kinds of Chinese medical materials totally according to statistics, in which 11,118 kinds are plant sources, 1,574 kinds are animal sources, and 80 kinds are mineral sources. These figures show that not only our natural Chinese medicinal resources are extremely rich, but also we are largely dependent on herbals. Especially in todays world, with the rapid development of society, the conflict between the needs of society and natural Chinese herbal resources becomes more and more prominent. There are many factors to affect the natural Chinese herbal resources, and the following four factors are particularly important. The first factor is that the application of Chinese herbs increases rapidly in population. Domestically, China's population was about 450 million at the founding of PRC, about 600 million at the sixties of last century, 1.16 billion at the fourth national census and 1.295 billion for the fifth national census. Meanwhile, along with the constant deepening of China's open policy, the radius of Chinese medicine is increased, and more and more people are using traditional Chinese medicine around the world, including more than 48 million overseas Chinese. In 2008, the amount of export of Chinese herbs exports were 1.3 billion US dollars and 163 countries traded with China in Chinese herbs, of which 154 are importing countries. Although the modern society doesnt use Chinese herbs just as simple as the ancient times, and chemical drugs have a large proportion of the application, the consumption of Chinese herbs is still at an increasing level because of the big figure of population. The second factor is non-medical consumption of Chinese herbs, which is the main factor increasing contradiction between the supply and demand of Chinese herbal resources. The purpose of the contemporary application of Chinese herbs has far exceeded the scope of treatment of the disease, and because of the incorrect publicity and the commercial interests, a myriad of different Chinese herbal industry are forming, such as health care, beauty, and medicated food. This huge consumer market competes with the Chinese medical market for the limited resources of Chinese herbs. The third factor is the expanding of the natural plant extract market. With the global

enthusiasm for natural medicines, and the rapid development of high and new technologies in plant chemistry, modern pharmaceutical industry, the research for finding new drug compounds or pro-drugs from the natural herbal is in the ascendant, and has gradually formed an industrial scale. Data shows that the consumption of plant extracts is about 10% of the total herbals. On the one hand, this is a development of drug research, but on the other hand, it is also a cruel plunder for Chinese herbal resources. For example, extraction rate of puerarin in pueraria is 3.58% (microwave-assisted), Mogroside V only 0.5% in fresh fruit, and Total Ginkgo Flavone-Glycoides only 0.15% in Ginkgo leaf, and the rest are discarded. It is true that, looking for new drug through the study of natural plant active ingredients is a very good idea, and in this regard bulbocapnine (1928-1936 Zhao Chenggu), ephedrine (1887 Nagai Nagayoshi) etc. are successful examples. However, the situation are changed now, a large number of manufacturing enterprises have mushroomed in various places, and is it a fortunate or disaster for sustainable development of the natural resources of Chinese herbs? The fourth factor is the combination of a number of factors, such as the changes of the natural environment and species, disorderly excavation and the international implementation of laws and regulations to protect animals and plants, which aggravates the situation of shortage of the Chinese herbal resources. In recent years, a large area of economic forestland cultivation was carried out, resulted in the damage and destruction of native plant resources to a huge degree. After many years collecting for industrial production, the wild species resources decline sharply, such as Shinyleaf Pricklyash Root and Liquorice Root, etc. For these mentioned reasons, many experts and scholars think deeply from macro to micro perspectives on how to meet the growing demand of the people and the international market for Chinese herbs, and to bring forth numerous ideas in favor of the sustainable development of the natural resources of Chinese herbs. Precisely at this time, we have proposed to carry out the idea of the medicinal study on agricultural residues, and the purpose of the study is to provide new prospects and approaches to the research of Chinese herbs, as well as increase additional resources of Chinese herbs. (2) The medicinal study on agricultural residues benefits the environment protection and promotes eco-agricultural development. The industry of Chinese herbal medicine is a resource-dependent industry. The demanding of Chinese herbs is growing, while the natural resources is decreasing, and the former will not be changed, so the only way is to change the latter in this contradiction.

Over the past 10 years, in order to solve this problem, with the implementation of modernization of Chinese herbal medicine, the state increased investment, build up several production bases of Chinese herbs, and a number of large pharmaceutical companies also chose suitable places of origin to establish their own production bases which are mainly related with their own products. Wanxi Pharmaceutical Co., Ltd invested a huge fund to build planting bases, each with an annual production of over 200 tons Chinese herbs across the country, such as Common Yam Rhizome base in Wuzhi County, Henan Province; Aahesive Rehmannia Root Tuber base in Wen County, Henan Province; Indian Buead and the root bark of the peony bases in Jinzhai County and Tongling County, Anhui Province and Oriental Waterplantain Tuber base in Jianou County, Fujian Province. Sanjiu Pharmaceutical Co.,Ltd has established 13 GAP planting bases of Upright Ladybell Root, Mongolian snakegourd, Indigowoad Root, Balloonflower Root, Ginseng, Safflower, Different Leaves Pseudostellaria Root Tuber, Thinleaf Milkwort Root, Rose, Aucklandia Lappa, Chinese Thorowax Root, Mongolian Milkvetch Root, Baikal Skullcap Root, etc. in Hebei, Anhui, Guizhou, Neimeng, Shandong Province etc.. Currently the number of planting bases of Chinese herbs is more than 1000 in our nation. Standards of the 500 bases are close to the national GAP certification standards. Planting area of Chinese herbs is about 21 million mu. A large-scale planting of Chinese herbs formed a new agriculture, and eased the contradiction between supply and demand of Chinese herbs to a certain extent. However, because China's land resources are extremely scarce and Chinese herbs planting takes up a large number of agricultural crops lands, the shortage of land resources is exacerbated. Thus, explore new herbal resources to find new uses or likely replace the existing species is an urgent and arduous task. We propose to carry out the medicinal study on agricultural residues, which is a measure to provide additional resources of medicinal plants without occupation of land resources. Moreover, a large number of waste products of crops are generated in agricultural production, and in the past, most of them were incinerated, or dumped in the fields and in the river directly, which caused serious environmental pollution. For example, annual production of sugar cane is about 50 million tons in Guangxi, and annual production of sugar cane leaves is about 7.5 million tons, except a small amount of leaves for cattle feeding, the vast majority is incinerated in situ. Again Guangxi is one of the largest provinces for mango cultivation, and just Baise Youjiang Valley alone, therere nearly 400,000 mu of land for mango growing, which results in over 200,000 tons mango leaves from pruning each year.

Tomato leaves, watermelon leaves and other staple crops are probably the same case. The burning of these waste products produces exhaust gas into the air, which results in decreased air quality and affects the ecological environment. Only the effective usage of these waste products can change this situation. (3) The medicinal study on agricultural residues promotes the development of circular economy and builds a harmonious society. The so-called circular economy is an efficient use and recycling of resources as the core of economic growth mode. From the perspective of resource utilization, the traditional economy is extensive and one-time, continuously turning the resources into the waste to achieve quantitative economic growth, while the circular economy promotes a kind of harmonious economic model with the environment, which organizes the economic activities into a feedback process of "resources - products - renewable resources". Reduce, reuse and recycle, is the "3R principle" of circular economy, in which recycling is divided into primary-level recycling and secondary recycling, and the medicinal study on agricultural residues proposed by us belongs to the secondary recycling, that is, "turning waste resources into raw material of other products " which is turning the waste products of crops into the raw materials of drugs or health products. To promote the development of circular economy in China, the National Development and Reform Commission and the State Environmental Protection Administration take a number of measures, which refers to "vigorously carrying out comprehensive utilization of resources, maximizing the use of resources, reducing the final disposal of waste, conducting comprehensive utilization of agricultural wastes generated in the process of production. In fact, according to the strategy of sustainable development since the 90s, developed countries are developing a circular economy, establishing recycling-based society as an important way and measure to implement sustainable development strategy. A number of pilot projects also conducted in the development of circular economy in China, but most of them are industrial-related projects, and especially none of them is related to turning the waste products of crops into medicinal resources in agriculture. In line with national development strategy of circular economy needs, the study on agricultural residues will enjoy broad application prospects. On the other hand, through modern science and technology, carrying out the study on agricultural residues to find the value and turn waste into treasure will not only solve the

problem of the environmental pollution caused by the original approach to dealing with such issues, but also increase the added value of crops for farmers to provide more employment opportunities, giving the local farmers new ways to shake off poverty and be on a road to prosperity. It can be said that this research project is necessary and feasible with remarkable economic and social benefits. Study of the mango leaf is a typical example. The early seventies of last century, Guangxi TCM University carried out the drug action research of mango leaves, and successfully developed "mango anti-cough tablets" with mango leaves as the main raw material. After nearly three decades of continuous development, there are four pharmaceutical companies and one hospital produce this kind of drug and the other hospital preparations with mango leaves as raw material nationally, and annual sales revenue has been more than 60 million Yuan. At least more than 10,000 tons of mango leaves are needed per year, and farmers in areas such as Baise have brought millions in revenue. This research is even more significant that it strongly extends and expands the industrial chain of mango cultivation, and becomes a bridge between the convergence of agriculture and industry, the fruit industry and pharmaceutical industry. 2. The general thoughts of the medicinal study on agricultural residues We propose to carry out the medicinal study on agricultural residues with the overall objective, i.e. to screen the functional ingredients of anti-inflammatory, anti-tumor,
anti-aging, lowering blood glucose and blood cholesterol etc. from mango leaves, sugar cane leaves, watermelon leaves, tomato leaves and some other staple crops, and to study the corresponding efficacy evaluation, to prove mechanism of action targets, to establish the active ingredient database of agricultural residues, to strive to find available medicinal resources from agricultural residues to solve problem of the increasing depletion of Chinese herbal resources and environmental pollution of waste products. To achieve this objective, we must focus on the four following measures at this stage and for a long period in the future. (1) To carry out the academic discussion of the medicinal study on agricultural residues to seek consensus and policy support The study on agricultural residues as new medicinal resources has a great significance in theory, but many difficulties in practice. First of all, it is the problem of understanding and policy. While most scholars and government officials have the attitude of appreciation, but people are still unfamiliar with it as a newborn of the medical academics; and it is impossible to show convincing efficiency except for the study of mango leaves because the basis of

preliminary studies of other agricultural residues is still very weak, and even literature are hard to find. This requires strong support of the academic community, and carrying out hard work to publicize and demonstrate the feasibility of the academic advocates by a variety of ways, not only form a consensus in the academic community, but more importantly urge the Government to give support to this project. Only being included in the national development of circular economy and ecological agriculture, will the protection of the sustainable development strategy of Chinese herbal resources be possible to carry out to achieve its intended objective. (2) Construction of the technology platform for the medicinal study on agricultural residues Technology platform is the basis of scientific research. Especially with regard to multidisciplinary research, there must be a technology integration platform which can guarantee that research goals. To carry out the medicinal study of waste products of crops, is a complicated systematic project, and from the disciplinary point of view, it is related to agriculture and medicine; From the industrial classification point of view, it is related to the primary industry of agriculture and the pharmaceutical industry of secondary industry, which breaks new ground for the future and links the two industries; From the perspective of science and technology it is related to modern biological information processing technology, plant chemistry (medicine chemical) technology, pharmaceutical technology, quality control and instrument analysis techniques, modern efficacy screening technology, food engineering, and so on. Therefore, research institutions with good basic conditions should be chosen. For example, in Guangxi, we can choose Baise National Agricultural Science and Technology Park and the Guangxi Research Center of Pharmacological Screening of Chinese Herbs. The former is a state-level agricultural science and technology research platform, and the latter is the provincial research science and technology platform of Chinese medicine. Such a functional Ingredients screening technology platform of agricultural residues, in which the research system, research team, technical equipments, operating mechanism, etc. should be designed based on the requirements of the international advanced level, and adequate financial support for scientific research should be provided in order to ensure the smooth development of research projects to strive to achieve model results within a short period of time. (3) To carry out civil investigations of the application of agricultural residues to develop long-term research programs

Although the study was recently proposed, but its object is crops which is closely related to peoples daily life, and the usage of crops are gradually accumulated and developed during the long agrarian times of the human society, and the theory " Medicinal and Edible" of Chinese herbs is built on this understanding of production and lifestyles. Therefore, there must have rich experiences applying crops waste to fight the disease in civil society, and this experiences maybe exist individually and scattered. These experiences will provide scientific research clues. The study of Mango leaves was derived from the Chinese herbal medicine movement at the early seventies of last century. In that movement, in their medical survey researchers found that local farmers have the habits of drinking the boiled water of mango leaves to treat cough, cholera illness in Bose and other places. According to this line of thought provided by civilian applications, the researchers conducted a series of study of mango leaves, resulting in the "mango anti-cough tablets, originally created in Guangxi. Therefore, we can say that civil application experiences of crops waste are sources to conduct this study for us. We build technology platforms and undertake a full investigation of the application of waste products of crops, including documents and on-site investigation. Based on the first-hand information, we develop the study plan and identify the short-term and long-term research objectives, tasks, specific targets and specific content, implementation steps, the progress and funding to ensure that research directions and goals are stable and feasible to avoid giving up halfway. (4) To form the research alliance of the medicinal study on agricultural residues Agriculture is the foundation of human society, even the most developed countries are also inseparable from the crops (just in different cropping patterns), and it is inevitable to produce agricultural residues. Take mango for an example, there are nearly 70 countries planting mango worldwide; and watermelon, tomato, etc. are also staple crops worldwide. In our country watermelons are planted from east to west and from south to north. In other words, the medicinal study on agricultural residues can and should become a worldwide international cooperation projects. In fact, during the systematic study of literature of mangiferin we found that the research of the mango leaves and mangiferin started earlier than China in the Western countries, especially deeply in basic research, but our work is more focus on applied research, and has made a lot of results especially in the clinical application and product development. The experts and scholars from various countries have their own advantages and characteristics in this area, so they should be combined to form a research
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alliance and set up two platforms: a platform for academic exchanges and another platform for technical support, so as to collaborate on common interest in a crop waste to establish a unified executable research program and share the work in accordance with their respective scientific and technological advantages. In this way, we will expect to make a landmark contribution to the field of medicinal study on agricultural residues in a short period of time.
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Study On Mango Leaf and Mangiferin Chemical Constituents with Unprecedented Skeletons from Alpinia katsumadai and Chukrasia tabularis var. velutina
Ling-Yi Kong Department of Natural Medicinal Chemistry, China Pharmaceutical University, 24 Tong Jia Xiang, Nanjing 210009, P. R. China E-mail: cpu_lykong@126.com
The seeds of Alpinia katsumadai Hayata (Zingiberaceae) have been used in traditional Chinese medicine (TCM) as an antiemetic agent and for the treatment of stomach disorders, and was coded in Chinese Pharmacopeia as an aromatic stomachic. From the petroleum ether extract of the seeds of A. katsumadai (20 Kg), a pair of unique sesquiterpenechalcone conjugates with unprecedented skeletons (1, 2)and two novel monoterpenechalcone conjugates (3, 4) was isolated, and the structure of 1, 2, 4 were confirmed by single-crystal X-ray diffraction. The stem bark of Chukrasia tabularis has been traditionally used as astringent, antidiarrheal, and anti-influenza agents in China. From the air-dried stem bark of C. tabularis var. velutina (10 Kg), nine novel phragmalin type limonoids were isolated. Three 16-norphragmalin limonoids, chukvelutins A-C (5-7), possess unprecedented skeletons featured with a characteristic ketal moiety between the phragmalin skeleton and a biosynthetically extended isobutyryl group at C-15. Six C-15-acyl phragmalin type limonoids, chukvelutilides A-F (8-13), are the first class of C-15-acyl phragmalin type limonoids with 16/30 -lactone ring confirmed by single-crystal X-ray diffraction.
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Study on Bioactive Compounds with Molecular Diversity from Toxic Plants in China
Shi-Shan Yu Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine, Ministry of Education; IMM, CAMS & PUMC, 100050, Beijing
Natural products play a dominant role in the discovery of leads for the development of drugs for the treatment of human diseases. In China, much of nature sources remain to be explored, particular the toxic plants, which leave no doubt that a host of novel, bioactive chemotypes await discovery. There are more than 900 species of toxic plants in our country. The bioactivities of extracts of over 150 toxic plants were investigated in our group. It was found that more than 20 toxic plants showed vasodilator activities and anti-tumor activities, of which 7 toxic plants were further studied by bioassay-guided technique. From the 7 toxic plants, more than 250 compounds were isolated, including 9 new skeleton compounds and more than 80 novel compounds, of which more than 50 compounds exhibited significant bioactivities to different targets. It lays a foundation for study on innovative drugs and elucidation of the bioactive substances of toxic plants.
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Pharmacology and Toxicology
Assessment of systemic interaction between swertia chirata extract and its bioactive constituents in rabbits
Satyendra Suryawanshi1, R. K. Asthana2 and R. C. Gupta1*
1 2
Pharmacokinetics and Metabolism Division, Central Drug Research Institute, Lucknow, India Medicinal and Process Chemistry Division, Central Drug Research Institute, Lucknow, India
The plant Swertia chirata (Gentianaceae) is known for its multifarious medicinal value in the Indian system of medicine (Ayurveda). Its methanol extracts having antidiabetic activity contains mangiferin, amarogentin, amaroswerin, sweroside and swertiamarin as active constituents. The pharmacokinetics of mangiferin and amarogentin have been carried out after intravenous administration of pure standards and extract from S. chirata (CT) in rabbits to assess systemic interaction. The remaining three components were also monitored in plasma for pharmacokinetic estimation based on the ratio analysis method. Mangiferin was characterized by a relative low clearance (~0.14L/h/kg) and a lesser volume of distribution (~0.15L/kg), while amarogentin exhibited a rapid clearance (~2.62L/h/kg) and wide distribution (~1.08L/kg) from the systemic circulation. No signicant difference was observed in pharmacokinetic parameters of mangiferin and amarogentin either administered alone or as CT formulation in rabbits.
The extraction of mangiferin from mango leaves and its analgesic function
Yingfang Wei,Lanyan Liao,Jie Lin,Qixin Lan,Guofeng Wei Department of Applied Chemistry, Youjiang Medical College for Nationalities, Baise, P.R. China
To investigate the extracting procedure of mangiferin from mango leaves and its analgesia, the organic solvents of ethanol and ethyl acetate were used repeatly to extract mango leaves. A kind of light yellow substance was obtained. The chemical quality analysis for this extracted substance was performed and the analgesia test in white mice (the mangiferin group) was also performed. All the results were analytically compared to those in aspirin and normal saline groups. The extracts mainly contained mangiferin. After analgesia test in small white mice, mangiferin could effectively relieve rats' pain induced by writhing, compared to aspirin and normal saline groups, there were statistically significant differences (all P<0.01).
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The mango leaves are abundantly contained mangiferin which has anagelsia gunction in rats.
The Effect of Kampo Formulae on Bone Resorption in Vitro and in Vivo. I Active Constituents of Tsu-kan-gan
Huiying Li1, Tatsuro Miyahara2, Yashuhiro Tezuka1, Tsuneo Namba1,Nobuo Nemoto2, Syuichi Tonami3, Hikaru Seto3, Takahiro Tada4, and Shigetoshi Kadota*,1
Research Institute for Wakan-Yaku (Traditional Sino-Japanese Medicines) ,Toyama Medical and
Pharmaceutical University,Toyama, Japan

2 3 4
Faculty of Pharmaceutical Science, Toyama Medical and Pharmaceutical University, Toyama, Japan Facluty of Medicine, Toyama Medical and Pharmaceutical University, Toyama, Japan Mikimoto Pharmaceutical Co.,Ltd., Ise, Japan
Four water extracts of Kampo formulae (Yi-kkan-sen, Dai-ho-in-gan, Ni-chi-gan, Tsu-kan-gan) were screened for their inhibitory activities on bone resorption induced by parathyroid hormone (PTH) in organ culture using neonatal mouse parietal bones. Among the Kampo formulae, Tsu-kan-gan(TKG) showed the most ptent inhibitory activity. We further fractionated the TKG water extract by monitoring the inhibitory on bone reportion stimulated by PTH in vitro. The MeOH fraction of the water extract inhibited PTH-stimulated bone resorption, and its inhibitory activity was more potent than thoses of other fractions. The MeOH fraction was then subiected to Sephadex LH-20 column chromatography to give fractions and
, which were examined for bone resorption activety. Fraction inhibited PTH-stimulated bone
resorption, and its inhibitory activeity was more potent than those of the other fractions. Upon oral administration of the three fractions (100 mg/kg/d) to ovariectomized(OVX) mice, fractions
and
prevented the decreased of bone mineral density (BMD) of the lumbar vertebra. Eleven compounds isolated from the MeOH fraction were examined for their inhibitory effect on PTH-stimulated bone resorption. Among them, berberine (1), syringin(3), limonin(4) and mangiferin(10) showed a significant inhibitory effect on bone resorption. In the formation assay of oseteoclast-like cells, these compounds decreased the number of tartarte-resistant acid phosphatase(TRAP)-positive multinucleated cells(MNCs). The inhibitory effect of TKG on bone resorption may be at least partly due to the inhibitory action of these compounds.
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Study On Mango Leaf and Mangiferin Mangiferin and hesperidin metabolites are absorbed from the
gastrointestinal tract of pigs after oral ingestion of a Cyclopia genistoides (honeybush tea) extract
Constance Bock1 ,Karl-Heinz Waldmann 2,Waldemar Ternes1*
1
Institute for Food Toxicology and Chemical Analytics, Center of Food Science, University of Veterinary
Medicine Hannover, Foundation, Hannover, Germany

2
Clinic fo Swine, Small Ruminants, Forensic Medicine and Ambulatory Service, University of Veterinary
Medicine Hannover, Foundation, Hannover, Germany
Health-promoting properties such as antioxidative, anticarcinogenic, and cholesterol-lowering Effects are described for mangiferin and hesperidin, the major phenolic compounds present in Cyclopia genistoides (honeybush). However, knowledge of their metabolic fate and their absorption from the gastrointestinal tract is very limited. The aim of this study was to determine the concentrations of mangiferin, hesperidin, and the irmetabolites in plasma, urine, and feces samples from pigs consuming an extract of Cyclopia genistoides. Pigs were administered upto 74 mg mangiferin perkilogram of body weight and 1 mg hesperidin per kilogram of body weight per day for 11 days. Plasma samples were collected at various time points on days 9 and 11 of the study and days 1 and 2 after termination of extract administration. Urine and feces were collected in fractions for 24 hours. In the plasma samples, the aglycone of mangiferin (norathyriol) was detected. Mean plasma concentrations ranged from 7.8 to 11.8 mol/L. Six metabolites of mangiferin and hesperidin were detected in the urine, including methyl mangiferin, norathyriol, itsmonoglucuronide, hesperetin, hesperetin monoglucuronide, and eriodictyol monoglucuronide. Between 26.0% and 30.8% of the administered dose of hesperidin and only between 1.4% and 1.6% of mangiferin could be detected in the urine on days 9 and 11 of the study. Approximately 8.2% of the administered dose of mangiferin was determined in the feces. The main metabolite was norathyriol. Neither hesperidin nor metabolites ascribed to hesperidin intake were detected. The results suggest that formation of norathyriol from mangiferin occurs in vivo, and specific metabolites were identified in blood and excretion products in urine and feces. This study wil laid ininvestigating the physiological functions of the parent compounds in vivo.
17
Pharmacokinetics of mangiferin in rat plasma after oral administration of a single dose of suanzaoren decoction
Li Yu-juan, BI Kai-ahun* School of Pharmacy, Shenyang Pharmaceutical University, Shenyang , China
To study the pharmacokinetics of mangiferin in rats after oral administration of a single dose of Suanzaoren decoction. An HPLC method was established using puerain as internal standard. Plasma samples were deproteinized with acetonitrile-acetic acid (9:1), followed by evaporation of the acetonitrile to dryness. The resultant residue was then dissolved in mobile phase and HPLC separation was achieved on a Hypersil C18 (200 mm 4.6 mm ID, 5 m) column at room temperature. The mobile phase consisted of acetonitrile-water (12: 88) with 1% acetic acid and 1% tetrahydrofuran at a flow rate of 0.7 mL min-1. The UV detection wavelength was set at 320 nm. The calibration curve was shown to be linear over the range from 0.536 to 26.8 g mL-1(r2 0.995). Mean recovery was determined as 92.7%. Within-day and between-day precisions were less than 9.1% RSD. The limit of quantitation (LOQ) was 0.536 g mL-1. The maximum plasma concentration (Cmax), the time to reach peak concentration (Tmax) and the apparent elimination half-life (T1/2) were (10.5 2.2) g mL-1, (5.8 0.4) h and (5.0 0.3) h, respectively. The validated HPLC method developed has been applied to take a limited view of pharmacokinetics profile of mangiferin in rat plasma after having orally taken a single dose of Suanzaoren decoction.
18
Study On Mango Leaf and Mangiferin Simultaneous estimation of mangiferin and four secoiridoid glycosides in rat plasma using liquid chromatography tandem mass spectrometry and its application to pharmacokinetic study of herbal preparation
Satyendra Suryawanshi1, R.K.Asthana 2,R.C.Gupta1*
1 2
Extracts from Swertia chirata (familyG entianaceae) have antidiabetics and antioxidant activity, largely attributed to the flavonoids and secoiridoids, which are a major class of functional components in methanolic extracts from aerial part of plants. In order to facilitate analysis of systemic exposure to S. chirata derived products in animals, we developed a liquid chromatography tandem mass spectrometry (LC-MS/MS) based method that is capable of routinely monitoring plasma levels of flavonoids and secoiridoids. An LC-MS/MS-based method has been developed for the simultaneous estimation of two bioactive markers, mangiferin and amarogention along with three other components, amaroswerin, sweroside and swertiamarin in rat plasma. All the analytes including the internal standard (kutkoside) were chromatographed on RP-18 column (250 mm4 mm i. d., 5m.) coupled with guard column using acetonitrile: 0.5 mM ammonium acetate buffer, pH3.0 as mobile phase at a flow rate of 1 ml/min in gradient mode. The final flow to source was splitted in 1: 1 ratio. The detection of the analytes was performed on API4000 LC-CMS/MS system in the multiplereaction-monitoring (MRM) mode. The quantitation for analytes other than the pure markers was based on relative concentration. The method was validated in terms of establishing linearity, specificity, sensitivity, recovery, accuracy and precision (Intra-and Inter-day), freeze-thaw stability, peltier stability, dry residue stability and long-term stability. There coveries from spiked control samples were >90% for all analytes and internal standard except mangiferin where recovery was >60%. Intra-and inter-day accuracy and precision of the validated method were within the acceptable limits of <15% at low and <10% at other concentrations. The quantitation method was successfully applied to generate pharmacokinetic (PK) profile of markers as well as to detect other components in plasma after intravenous dose administration of herbal preparation in male Sprague-Dawley (SD) rats.
19
Study On Mango Leaf and Mangiferin UV/vis, 1H, and pKa values
Berenice
13
C NMR spectroscopic studies to determine mangiferin
Gmez-Zaleta1,
Mara
Teresa
Ramrez-Silva1,
Atilano
Gutirrez1,
Enrique
Gonzlez-Vergara2, Marisol Guizado-Rodrguez3, AlbertoRojas-Hernndez1*

1
Universidad Autonoma Metropolitana-Iztapalapa, Depto. Qumica, Areade Qumica Analtica, Apdo. Postal 55-534, 09340 Mxico DF, Mxico
2 3
Centro de Qumca, Instituto de Ciencias, Benemrita Universidad Autonoma de Puebla, Mxico Depto. De Qumica Inorgnica, Instituto de Qumica, UNAM, Ciudad Universitaria, Mxico DF, Mexico
The acid constants of mangiferin (a natural xanthonoid) in aqueous solution were determined through an UV/vis spectroscopic study employing the SQUAD program as a computational tool. A NMR study complements the pKa values assignment and evidences a Hbridge presence on 1-C. The chemical model used was consistent with the experimental data obtained. The pKa values determined with this procedure were as follows:H4 = (MGF)-+ H+, pKa1 (6-H) =6.520.06; H 3 (MGF) - = H 2 (MGF) 2-+H, pKa2 (3-H) = 7.97 0.06; H 2(MGF)
2-
= (MGF) 3 - + H+, pKa3 (7-H)=9.440.04; H(MGF) 3-= (MGF) 4-+H+ ,
pKa4 (1-H)=12.100.01; where it has been considered Mangiferin C19H18O11 as H4(MGF). Mangiferin UV/vis spectral behavior, stability study in aqueous solution as well as NMR spectrocopy studies: one-dimensional
1
H,
13
C,2D correlated 1H/
13
C performed by (g)-HSQC and (g)-HMBC methods; are also presented. pKa in aqueous solution is a necessary contribution to subsequent
values determination of H4(MGF)
pharmacokinetic study, and a step towards the under standing of its biological effects.
20
Study On Mango Leaf and Mangiferin A review of the bioactivity of south african herbal Teas: rooibos (aspalathus linearis) and honeybush (Cyclopia intermedia)
Diane L. McKay* and Jeffrey B. Blumberg USDA Human Nutrition Research Center on Aging at Tufts University, Boston, USA
Rooibos (Aspalathus linearis) and honeybush (Cyclopia intermedia) are popular tisanes in their native South Africa and have a growing worldwide market. Both herbal teas are used traditionally for medicinal purposes and are rich in polyphenols with rooibos a rare source of the dietary dihydrochalcones, aspalathin and nothofagin.The principal polyphenols in honeybush include the xanthone mangiferin and the flavonones hesperitin and isokuranetin. Despite their divergent phytochemical and nutrient compositions, rooibos and honeybush share potent antioxidant and antimutagenic activities in vitro. Animal model studies indicate both herbal teas possess potent antioxidant, immune-modulating and chemopreventive actions. However, human studies ofrooibos are limited and of honeybush are absent. No adverse effects of rooibos or honeybush consumption astisanes have been reported.
Activation of lymphocytes of normal and tumor bearing mice by mangiferin, a naturally occurring glucosylxanthone
Utpala Chattopadhyay1, Swatidas1, surajit Guha1 ,Shibnath ghosal2
1 2
Department of Tumor Immunobiology, chittaranjan National Cancer Research Centre, Calcutta,India Department of Pharmaceutics, Institute of Technology, Banaras Hindu University, Varanasi,Iindia
Mangiferin, a naturally occurring glucosylxanthone, was assessed for its immunomodulatory potential. The phytochemical induced extensive in vitro proliferation of murine splenocytes and thymocytes at the doses of 5-40 gml-1. Suppression of the proliferative response of the cells was observed with higher doses of mangiferin. Mangiferin also activated the splenocytes of tumor hosts at early and late stages of tumor growth. The Phytohemagglutinin (PHA) and Con A unresponsive plenocytes of advanced tumor bearer proliferated extensively in response o mangiferin. Mangiferin when used with Con A produced additive stimulatory effect and induced heightened DNA synthesis of bearers splenocytes. normal and advanced tumor
21
Study On Mango Leaf and Mangiferin An Anacardiaceae preparation reduces the inflammation-related genes in murine macrophages
J . Leiro1*, D.
1
expression
of
Garca2, J. A. Arranz1, R. Delgado2, M. L. Sanmartn1, F. Orallo3
Laboratorio de Parasitologa, Instituto de Investigaciones y Anlisis Alimentarios, Universidad de Santiago de Compostela, Constantino Candeira, sn, Santiago de Compostela, Spain.
Laboratorio de Farmacologia, Centro de Quimica Farmaceutica, Havana, Cuba Departamento de Farmacologia, Facultad de Farmacia, Universidad de Santiago de Compostela, Santiago de Compostela, Spain
This study investigated the effects of an aqueous extract of the stem bark of Mangifera indica L. (Anacardiaceae; Vimang), which contains a defined mixture of components including polyphenols (principally mangiferin, MA), triterpenes, phytosteroids, fatty acids and microelements, on expression of inflammation mediators in inflammatory murine macrophages after stimulation in vitro with lipopolysaccharide (LPS) and interferon-gamma (IFN-). In vitro treatment with Vimang at 4 microg/ml reduced levels of NOS-2 mRNA and NOS-2, while treatment at 40 g/ml also reduced levels of COX-2 mRNA, COX-2, and prostaglandin E2 (PGE2). Results suggested that MA is involved in these effects. In vitro treatment with Vimang at 40 microg/ml also inhibited mRNA levels of the proinflammatory cytokines interleukin 1beta (IL-1), tumor necrosis factor alpha (TNF-) and colony-stimulating factor (GM-CSF), but did not affect mRNA levels of IL-6 or tumor growth factor-beta (TGF-). Extracellular release of TNF- by inflammatory macrophages was inhibited by in vitro treatment with Vimang at the same concentrations that showed inhibition of TNF- mRNA levels. The inhibition of TNF- production appears to be at least partially attributable to MA. Vimang at 4 microg/ml decreased mRNA levels of nuclear factor-kappaB (NF-B) but did not affect expression of the NF-B inhibitor (IB). These data indicate that the potent anti-inflammatory effects of Vimang are due to selective modulation of the expression of inflammation-related genes, leading to attenuation of macrophage activation.
22
Study On Mango Leaf and Mangiferin Anthelminthic and antiallergic activities of Mangifera indica L. Stem bark components vimang and mangiferin
D. Garc1, M. Escalante2, R. Delgado2, F. M. Ubeira2 and J. Leiro3*
1 2
Laboratorio de Farmacologa, Centro de Qumica Farmacutica, Havana, Cuba Laboratorio de Parasitologa, Facultad de Farmacia, Universidad de Santiago de Compostela, Santiago de Compostela, Spain
Laboratorio de Parasitologa, Institute de Investigaciny Aulisis Alimentarios, Universidad de Santiago de Compostela, Santiago de Compostela, Spain
This study investigated the antiallergic and anthelmintic properties of Vimang (an aqueous extract of Mangifera indica family stem bark) and mangiferin (the major polyphenol present in Vimang) administered orally to mice experimentally infected with the nematode, Trichinella spiralis. Treatment with Vimang or mangiferin (500 or 50 mgkg-1 body weight per day, respectively) throughout the parasite life cycle led to a signicant decline in the number of parasite larvae encysted in the musculature; however, neither treatment was effective against adults in the gut. Treatment with Vimang or mangiferin likewise led to a signicant decline in serum levels of specic anti-Trichinella IgE, throughout the parasite life cycle. Finally, oral treatment of rats with Vimang or mangiferin, daily for 50 days, inhibited mast cell degranulation as evaluated by the passivecutaneous anaphylaxis test (sensitization with infected mouse serum with a high IgE titre, then stimulation with the cytosolic fraction of T. spiralis muscle larvae). Since IgE plays a key role in the pathogenesis of allergic diseases, these results suggest that Vimang and mangiferin may be useful in the treatment of diseases of this type.
23
Study On Mango Leaf and Mangiferin Anti-allergic properties of Mangifera indica L. extract (Vimang) and contribution of its glucosylxanthone mangiferin
Dagmar Garca Rivera , Ivones Hernndez Balmaseda , Alina Alvarez Len , Belkis C ancio Hernndez , Luca Mrquea Montiel , Gabino Garrido Garrido , Salvatore Cuzzocrea , Ren Delgado Hernndez Laboratory of Pharmacology, Department of Biomedical Research, Center of Pharmaceutical Chemistry, Havana, Cuba.
Vimang is the brand name of formulations containing an extract of Mangifera indica L., ethnopharmacologically used in Cuba for the treatment of some immunopathological disorders, including bronchial asthma, atopic dermatitis and other allergic diseases. However, the effects of Vimang on allergic response have not been reported until now. In this study, the effects of Vimang and mangiferin, a C-glucosylxanthone isolated from the extract, on different parameters of allergic response are reported. Vimang and mangiferin showed a significant dose-dependent inhibition of IgE production in mice and anaphylaxis reaction in rats, histamine-induced vascular permeability and the histamine release induced by compound 48/80 from rat mast cells, and of lymphocyte proliferative response as evidence of the reduction of the amount of B and T lymphocytes able to contribute to allergic response. In these experiments, ketotifen, promethazine and disodium cromoglicate were used as reference drugs. Furthermore, we demonstrated that Vimang had an effect on an in-vivo model of inflammatory allergy mediated by mast cells. These results constitute the first report of the anti-allergic properties of Vimang on allergic models, as well as suggesting that this natural extract could be successfully used in the treatment of allergic disorders. Mangiferin, the major compound of Vimang, contributes to the anti-allergic effects of the extract.
24
Study On Mango Leaf and Mangiferin Antidiabetic activity of a xanthone compound, mangiferin
Toshihiro Miura1, Hiroyuki Ichiki2, Itsuko Hashimoto1, Naoki Iwamoto1, Motoshi Kato1, Masayoshi Kubo2, Eriko Ishihara1, Yasuhiro Komatsu2, Minoru Okada2, Torao Ishida and Keiichro Tanigawa1.
1 2 3
Suzuka University of Medical Science, Suzuka, Mie, Japan Hi-tech Research Center, Suzuka University of Medical Science, Suzuka, Mie, Japan Tsumura Central Research Laboratories, Tsumura and Co., Ami-machi, Inashiki-gun, Ibaraki, Japan
Mangiferin (MF) isolated from Anemarrhena asphodeloides Bunge rhizome, was tested forantidiabetic activity in KK-Ay mice, an animal model of type-2 diabetes. MF lowered the blood glucose level of KK-Ay mice 3 weeks after oral administration ( p< 0.01). However, no effect on the blood glucose level in normal mice was seen, indicating that MF could be useful in treating type-2 diabetes. In addition, MF improved hyperinsulinemia and, on insulin tolerance test, reduced blood glucose levels of KK-Ay mice. From these findings, it seems likely that MF exerts its antidiabetic activity by decreasing insulin resistance.
Antidiabetic activity of the rhizoma of anemarrhena asphodeloides and active components, mangiferin and its glucoside
Toshihiro Miura1*, Hiroyuki Ichiki3, Naoki Iwamoto1, Motoshi Kato1, Masayoshi Kubo3,Hiroshi Sasaki3, Minoru Okada3, Torao Ishida4, Yutaka Seino2, and Keiichiro Tanigawa1
1 2 3 4
Department of Clinical Nutrition, Suzuka University of Medical Science, Suzuka, Japa, Department of Metabolism and Clinical Nutrition, Kyoto University School of Medicine, Kyoto, Japan Tsumura Central Research Laboratories, Tsumura and Co.c 3586 Yoshiwara, Amimachi, Ibaraki, Japan Hi-tech Research Center, Suzuka University of Medical Science Kishioka, Mie,Japan
The antidiabetic activity of the rhizoma of Anemarrhena asphodeloides was investigated in KK-Ay mice, an animal model of genetic type 2 diabetes. The water extract of the rhizoma (AA) (90mg/kg) reduced blood glucose levels from 57029 to 40159mg/dl 7h after oral administration (p0.05) and also tended to reduce serum insulin levels in KK-Ay mice. AA-treated KK-Ay mice had significantly reduced blood glucose levels in an insulin tolerance test. Based on these results, the antidiabetic mechanism of AA may be due to decreased insulin resistance. In addition, the active components of AA were confirmed to be mangiferin and its glucoside.
25
Study On Mango Leaf and Mangiferin Antiinflammatory, analgesic and hypoglycemic effects of Mangifera indica Linn. (Anacardiaceae) stem-bark aqueous extract
J.A.O. Ojewole Department of Pharmacology, Faculty of Health Sciences, University of KwaZulu-Natal, Durban, South Africa.
Previous studies in our laboratories and elsewhere have shown that some members of Anacardiaceae family possess antiinflammatory, analgesic and hypoglycemic effects in man and mammalian experimental animals. The present study was, therefore, undertaken to examine the antiinflammatory, analgesic and antidiabetic properties of the stem-bark aqueous extract of Mangifera indica Linn., M. indica a member of the Anacardiaceae family, in rats and mice. The stem-bark powder of M. indica was Soxhlet extracted with distilled water and used. The analgesic effect of the plant's extract was evaluated by the hot-plate and acetic acid test models of pain in mice, while the antiinflammatory and antidiabetic effects of the stem-bark extract were investigated in rats, using fresh egg albumin-induced paw edema, and streptozotocin (STZ)-induced diabetes mellitus, respectively. Morphine (MPN, 10 mg/kg i.p.), diclofenac (DIC, 100 mg/kg i.p.), and chlorpropamide (250 mg/kg p.o.) were used respectively as reference analgesic, antiinflammatory, and hypoglycemic agents for comparison. M. indica stem-bark aqueous extract (MIE, 50-800 mg/kg i.p.) produced dose-dependent and significant (p<0.05-0.001) analgesic effects against thermally and chemically induced nociceptive pain stimuli in mice. MIE (50-800 mg/kg i.p.) also significantly (p<0.05-0.001) inhibited fresh egg albumin-induced paw edema, and caused significant (p<0.05-0.001) hypoglycemic effects in rats. It is suggested that the analgesic effects of MIE (50-800 mg/kg i.p.) may be peripherally and centrally mediated. The different chemical constituents of the plant, especially the polyphenolics, flavonoids, triterpenoids, mangiferin, and other chemical compounds present in the plant may be involved in the observed antiinflammatory, analgesic, and hypoglycemic effects of the plant's extract. However, the results of this experimental animal study lend pharmacological credence to the suggested folkloric uses of the plant in the management and control of painful, arthritic and other inflammatory conditions, as well as in the management of adult-onset type 2 diabetes mellitus in some rural African communities.
26
Study On Mango Leaf and Mangiferin Antitumor, immunomodulatory and anti-HIV effect of mangiferin, a naturally occurring glucosylxanthone
Surajit Guha1 , Shibnath Ghosal2, Utpala Chattopadhyay1
1
Department of Tumor Immunobiology, Chittaranjan National Cancer Institute, Calcutta Department of Pharmaceutics Institute of Technology, Banaras Hindu University, Varanasi, India
Mangiferin,a C-glucosylxanthone (1,3,6,7-tetrahydroxyxanthone-C2-beta-D-glucoside) purified from plant sources was shown to have in vivo growth-inhibitory activity against ascitic fibrosarcoma in Swiss mice. Following in vivo or in vitro treatment, it also enhanced tumor cell cytotoxicity of the splenic cells and peritoneal macrophages of normal and tumor-bearing mice. In vitro treatment of the splenic cells of tumor-bearing mice with mangiferin resulted in augmented killing of tumor cells, both resistant and sensitive to natural killer cells. Mangiferin was also found to antagonize in vitro the cytopathic effect of HIV. The drug appears to act as a potent biological response modifier with antitumor and antiviral effect.
Chemopreventive efficacy of mangiferin against benzo(a)pyrene induced lung carcinogenesis in experimental animals
Peramaiyan Rajendran, Ganapathy Ekambaram, Venkatraman Magesh, Dhanapal Sakthisekaran* Department of Medical Biochemistry, Dr. ALM PG Institute of Basic Medical Sciences, University of Madras, Taramani Campus, Chennai, India
Chemoprevention has emerged asavery effective preventive measure against carcinogenesis. Several bioactive compounds present in fruits and vegetables have revealed their cancer curative potential on carcinogenesis. Tumor markers correlate strongly with prognosis on tumo rburden. Glycoprotein and membrane ATPases play an important role in carcinogenesis. Hence this study was launched to evaluate the effect of mangiferin on the changes in glycoprotein components, ATPases and membrane lipidp eroxidation in control and lung carcinoma bearing mice. Asignificant increase in the levels of glycoproteins, membrane ATPases and membrane lipid peroxidation were observed in animals with lung carcinoma. On administration of mangiferin, these changes were reverted back to near normal levels. The increased levels of glycoprotein components found in lung carcinoma were also significantly decreased in mangiferin treated. Overall, the above data shows that the anticancer effect of mangiferin is more pronounced when used as an chemopreventive agent rather than as a chemotherapeutic agent against B(a)P induced lung carcinogenesis.
27
Study On Mango Leaf and Mangiferin -D-Glucoside suppresses tumor necrosis factor-induced activation of uclear transcription factor B but potentiates apoptosis
Abira Sarkar1*, Yashin Sreenivasan1, Govindarajan T. Rame2 ,and Sunil K. Manna1
1 2
Centre for DNA Fingerprinting & Diagnostics, Laboratory of Immunology, Hyderabad,India Department of Biology, Texas Southern University, Houston, Texas,U.S.A.
Mangiferin,
natural
polyphenol
is
known
to
exhibit
anti-inflammatory,
antioxidant,
andantiviraleffects. However the molecular mechanism underlying these effects has not been well characterized. Because NF-B plays an important role in these processes, it is possible that mangiferin modulates NF-B activation. Our results show that mangiferin blocks tumor necrosis factor (TNF)-induced NF-B activation and NF-B-dependent genes like CAM1 and COX2. The effect was mediated through inhibition of IKK activation and subsequent blocking of phosphorylation and degradation of IB. In addition, mangiferin inhibits TNF-induced p65 phosphorylation as well as translocation to the nucleus and also inhibits NF-B activation induced by other inflammatory agents like PMA, ceramide, and SA-LPS. Mangiferin, similar to the other known antioxidants, NAC and PDTC, inhibits TNF-induced reactive oxygen inter mediate (ROI) generation. Since intracellular glutathione (GSH) levels are known to modulate NF- B levels, we measured the levels of GSH. Mangiferin enhances glutathione level by almost 2-fold more than other antioxidants, and at the same time it decreases the levels of GSSG and increases the activity of catalase. Depletion of GSH by buthionine sulfoximine led to asignificant reversal of mangiferin effect. Hence mangiferin withits ability to inhibit NF-B and increase the intracellular GSH levels ma prove to be a potent drug for anti-in-flammatory and antioxidant therapy. Mangiferin-mediated down-regulation of NF-B also potentiates chemotherapeutic agent-mediated cell death, suggesting a role in combination therapy for cancer.
28
Study On Mango Leaf and Mangiferin Cytoprotective and antigenotoxic potential of Mangiferin, a glucosylxanthone against cadmium chloride induced toxicity in HepG2 cells
B. S. SatishRao 1*, M. V. Sreedevi2 , B. Nageshwar Rao1
1
Division of Radiobiology and Toxicology, Manipal Life Sciences Centre, Manipal University, Manipal, Karnataka,India
Division of Biotechnology,Manipal Life Sciences Centre,Manipal,Manipal University,Manipal, Karnataka,India
Mangiferin (MGN), a glucosylxanthone present inlarge amounts in the leaves and edible mango fruits of Mangiferaindica. Here, we report about MGNs potential fo rmitigating cadmium chloride (CdCl2) induced cytotoxic and genotoxic effects in HepG2 cells growing invitro. The cytoprotective potential was assessed by MTT, clonogenic and apoptotic assays, while antigenotoxic effect by micronucleus and cometassay. The established cytotoxic and genotoxic effects were well indicated after CdCl2 treatment and was mitigated by pretreatment with MGN. MGN prior to CdCl2 treatment invreased the cell survival (MTT), surviving fraction (clonogenicassay) and inhibited sub-G1 population (flow cytometric analysis) . Further, inhibition of CdCl2 induced apoptotic cell death by MGN was confirmed by microscopic and DNA fragmentation assays. Asignificant (p<0.01) reduction in the micronuclei frequency and comet parameters after MGN pretreatment to CdCl2 Clearly indicated the antigenotoxic potential. Similarly, the reactive oxygen species generated by the CdCl2 treatment were inhibited significantly (p<0.001) by MGN. Taken together, our study revealed that MGN has potent cytoprotective and antigenotoxic effect against CdCl2 induced toxicity in HepG2 cell line and which may be attributed to decrease in CdCl2 induced reactive oxygen species levels and resultant oxidative stress.
29
Study On Mango Leaf and Mangiferin Cytoprotective effect of mangiferin on benzo(a) pyrene-induced lung carcinogenesis in swiss albino mice
Peramaiyan Rajendran, Ganapathy Ekambaram and Dhanapal Sakthisekaran Department of Medical Biochemistry, Dr. ALM PG Institute of Basic Medical Sciences, University of Madras, Taramani Campus, Chennai, India
Antioxidants are one of the key players in tumourigenesis, and several natural and synthetic antioxidants have been shown to have anticancer effects. In the present investigation, the efficacy of mangiferin on the antioxidant status of benzo(a) pyrene-induced lung carcinogenesis in Swiss albino mice was assessed. The animals were divided into five groups. The animals in groups I and V were normal control and mangiferin control, respectively. Groups II, III and IV were administered with benzo(a) pyrene (50 mg/kg body weight, orally) for 4 weeks (twice a week) to induced lung carcinogenesis. Starting 1 week prior to benzo(a) pyrene administration, group III animals were treated with mangiferin (100 mg/kg body weight) in the diet for 18 weeks; 12 weeks after benzo(a) pyrene administration, group III animals were treated with mangiferin that continued until the end of the experiment period (18 weeks). At the end of the experiment period, the reactive oxygen species, glutathione and the activities of antioxidant enzymes were assessed in both lung and liver tissues. The levels of glutathione, superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, vitamin E and vitamin C were decreased in group II animals. However, in the mangiferin + benzo(a) pyrene-treated groups III and IV, the levels of GSH and the activities of antioxidant enzymes in both lung and liver were improved when compared with benzo(a) pyrene-induced group II animals. In addition, the finding that mangiferin decreased reactive oxygen species levels and enhanced antioxidant status suggests that this polyphenol might also be of value in the prevention of benzo(a) pyrene-induced lung carcinogenesis.
30
Study On Mango Leaf and Mangiferin Differential oxidative stress in oligodendrocytes and neurons after excitotoxic insults and protection by natural polyphenols
Gaskon Ibarretxe , Mara Victorla Snchez-Gmez, Mara Rosario Campos-Esparza , Elena Alberdi , Carlos Matute Departamento de Neurociencias, Facultad de Medicina y Odontologia, Universidad del Pas Vasco, Leioa, Spain.
Oligodendrocytes are vulnerable to overactivation of both their AMPA receptors and their high- and low-affinity kainate receptors. Depending on the intensity of the insult and the type of receptor activated, excitotoxic oligodendrocyte death mediated by these receptors has different characteristics. One important consequence at a cellular level is the ensuing oxidative stress, related to Ca2+-dependent alterations in mitochondrial functioning. We observed that oxidative stress associated with selective AMPA receptor activation is much higher than that associated with the selective activation of high- and low-affinity kainate receptors. Moreover, excitotoxic insults generate more intense oxidative stress in oligodendrocytes than in cortical neurons, though similar alterations in [Ca2+] and mitochondrial potential were observed in both cell types. Nanomolar concentrations of mangiferin and morin, two natural polyphenols with antioxidant properties, partially protect oligodendrocytes as well as cortical neurons from mild, but not intense, insults mediated by AMPA receptors. In addition to presenting oxygen radical scavenging activity, mangiferin and morin attenuate the intracellular Ca2+ overload subsequent to the activation of AMPA receptors, a mechanism that may contribute to their protective properties. The inclusion of these antioxidant agents in therapeutic strategies for the treatment of diseases in which oligodendrocyte as well as neuron loss occurs may prove to be beneficial.
31
Study On Mango Leaf and Mangiferin Dual mechanism of mangiferin protection against iron-induced damage to 2-deoxyribose and ascorbate oxidation
Gilberto Lzaro Pardo-Andreu1,2* , Ren Delgado3, Alberto J. Nez-Sells3, Anibal E. Vercesi 1
1
Departamento de Patologia Clnica, Faculdade de Cincias Mdicas, Universidade Estadual de Campinas, Campinas, SP, Brazil
2 3
Departamento de Farmacia, Universidad de Camaguey, Carretera Circunvalacion, Camaguey, Cuba Departamen to deInvestigaciones Biomedicas, Centrode Qumica Farmaceutica, Habana, Cuba
We studied mangiferin effects on the degradation of 2-deoxyribose induced by Fe(III) -EDTA/citrate plus ascorbate, in relation to ascorbate oxidation (measured at 265 nm). Results revealed that mangiferin was equally effective in preventing degradation of both 15 and 1.5 mM 2-deoxyribose. At a fixed Fe (III) concentration, increasing the concentration of ligands (either EDTA or citrate) caused a significant reduction in the protective effects of mangiferin. Interestingly, mangiferin strongly stimulated Fe(III) -EDTA ascorbate oxidation, but inhibited it when citrate was used as iron co-chelator. Mangiferin stimulated O2 consumption due to Fe (II) (formed by Fe (III) ascorbate reduction) autoxidation when the metal ligand was EDTA, but inhibited it when citrate was used. These results suggest that mangiferin removes iron from citrate, but not from EDTA, forming an iron-mangiferin complex that cannot induce ascorbate oxidation effectively, thus inhibiting iron-mediated oxyradical formation. Taken together, these results indicate that mangiferin works mainly by a mechanism different from the classical hydroxyl radical scavengers, keeping iron in its ferric form, by complexing Fe (III), or stimulating Fe (II) autoxidation.
32
Study On Mango Leaf and Mangiferin Effect of Mangifera indica L. extract (QF808) on protein and hepatic microsome peroxidation
Gregorio Martnez 1*, Attilia Giuliani 2, Olga Sonia Len1 , Gema Prez 1, Alberto J Nez Selles3
1
Center for Evaluation and Biological Research. Institute of Pharmacy, Havana University, San Lzaro y L, Havana 4, Cuba.
2 3
Department of Chemistryand Medical Biochemistry, viaSaldini, Milan, Italy Center of Pharmaceutical Chemistry, Havana, Cuba
The antioxidant activities of QF808, a steam bark extract of Mangifera indica L., were studied on hydroxyl-mediated oxidation of bovine serum albumin (BSA) and in a hepatic microsome system. The extract was effective in reducing the oxidation of BSA, since its half- maximal inhibition concentration (IC50 was 0.0049% w/v in the inhibition of carbonyl group formation and lower than 0.0025% w/v in the inhibition of sulfhydryl group loss. QF808 inhibited lipid peroxidation which was initiated enzymatically by reduced nicotinamide adenine dinucleotide phosphate (NADPH), IC50=0.00075% w/v, or non-enzymatically by ascorbic acid, IC50=0.0126% w/v. The extract tested did not inhibit NADPH-dependent cytochrome P-450 reductase activity, since it had no effect on the oxidation rate of NADPH. These results suggest that QF808 has an antioxidant activity, probably due to its ability to scavenge free radicals involved in microsome lipid peroxidation. In addition, QF808 antioxidant profile in vitro is probably similar to its principal polyphenolic component, mangiferin, a glycosylated xanthone.
33
Study On Mango Leaf and Mangiferin Effect of mangiferin on benzo(a)pyrene induced lung carcinogenesis in experimental Swiss albino mice
P. Rajendran , G. Ekambaram , D. S akthisekaran * Department of Medical Biochemistry, Dr. ALM PG Institute of Basic Medical Sciences, University of Madras, Chennai, India.
The present study is an effort to identify a potent chemopreventive agent against cancer, in which oxidative stress plays an important causative role. The modulatory effect of mangiferin on mitochondrial lipid peroxidation (LPO), tricarboxylic acid (TCA) cycle key enzymes and electron transport chain complexes was investigated against lung carcinogenesis induced by benzo(a)pyrene (50 mgkg-1 b/w orally) in Swiss albino mice. Decreased activities of electron transport chain complexes and TCA cycle key enzymes such as isocitrate dehydrogenase (ICDH), succinate dehydrogenase (SDH), malate dehydrogenase (MDH) and alpha-ketoglutarate dehydrogenase (-KGDH), in lung cancer bearing animals were observed. Pre- and post-treatment with mangiferin (100 mgkg-1 b/w orally) for 18 weeks, prevented the above biochemical changes, which were inclined towards normal control animal values. This study further confirms the chemopreventive and chemotherapeutic effect of mangiferin and these results are consistent with our hypothesis that mangiferin is a promising chemopreventive agent.
Efficacy of mangiferin on serum and heart tissue lipids in rats subjected to isoproterenol induced cardiotoxicity
Prabhu Sukumaran Nair* , C.S. Shyamala Devi Department of Biochemistry, University of Madras, Guindy Campus, Chennai, India.
The efficacy of mangiferin, on metabolism of lipids was tested in experimental cardiotoxic rats. The cardiotoxicity was induced by myocardial infarction through subcutaneous administration of isoproterenol hydrochloride for 2 days using 0.1 ml saline. Mangiferin drug was given as pretreatment for 28 days through intraperitonial administration using 0.2 ml dimethyl sulphoxide. Mangiferin significantly reduced the cholesterol, triglycerol, free fatty acids levels in serum and heart of the cardiotoxic myocardial infarcted rats. Mangiferin also increased the level of heart tissue phospholipids significantly in isoproterenol induced cardio toxic rats. The experiment thus concludes that mangiferin possess cardioprotective and hypolipidemic effect on experimentally induced cardiotoxic myocardial infarcted rats.
34
Study On Mango Leaf and Mangiferin Effect of mangiferin on hyperglycemia and atherogenicity in streptozotocin diabetic rats
S. Muruganandan 1*, K. Srinivasan 2, S. Gupta1 , PK Gupta 1, J. Lal 1
1 2
Division of Pharmacology and Toxicology, Indian Veterinary Research Institute, Izatnagar, India Department of Pharmacology and Toxicology. National Institute of Pharmaceutical Education and Research. Mohali. Punjab. India
In the present study, the effect of mangiferin (a xanthone glucoside, isolated from the leaves of Mangifera indica) on the atherogenic potential of streptozotocin (STZ)-diabetes was investigated. In addition, the effect of mangiferin on oral glucose tolerance in glucose-loaded normal rats was also determined. The chronic intraperitoneal (i.p.) administration of mangiferin (10 and 20 mg/kg) once daily (o.d.) for 28 days exhibited antidiabetic activity by significantly lowering fasting plasma glucose level at different time intervals in STZ-diabetic rats. Further, mangiferin (10 and 20 mg/kg, i.p.) showed significant antihyperlipidemic and antiatherogenic activities as evidenced by significant decrease in plasma total cholesterol, triglycerides, low-density lipoprotein cholesterol (LDL-C) levels coupled together with elevation of high-density lipoprotein cholesterol (HDL-C) level and diminution of atherogenic index in diabetic rats. In addition, the chronic administration of mangiferin (10 and 20 mg/kg, i.p.) for 14 days significantly as well as markedly improved oral glucose tolerance in glucose-loaded normal rats suggesting its potent antihyperglycemic activity. The accumulating evidences suggest that both pancreatic and extrapancreatic mechanisms might be involved in its antidiabetic or antihyperglycemic action. In conclusion, the present study demonstrates that mangiferin possesses significant antidiabetic, antihyperlipidemic and antiatherogenic properties thus suggesting its beneficial effect in the treatment of diabetes mellitus associated with hyperlipidemia and related cardiovascular complications.
35
Study On Mango Leaf and Mangiferin Effect of mangiferin on mitochondrial energy production in experimentally induced myocardial infarcted rats
S. Prabhu , Mallika Jainu , K.E. Sabitha , C.S. Shyamala Devi * Department of Biochemistry, University of Madras, A.C. Tech., Guindy campus, Chennai, India.
Mangiferin, from the leaves of Mangifera indica Linn., has been suggested as useful in the treatment of cardiovascular disorders. In the present study this drug was examined on the alteration of cardiac energy metabolism in isoproterenol (ISPH) administered myocardial infarcted rats. ISPH (20 mg/kg b.w.), which was administered s.c. twice at an interval of 24 h, caused a significant decrease in the activities of TCA cycle enzymes and antioxidant defense enzymes with a concomitant increase in the lipid peroxidation of heart mitochondria in rat model. The ATP production and the oxidation of succinate in State 3 and 4 decreased significantly in the cardiac mitochondria of ISPH administered rats. These functional alterations were supported by severe modifications in mitochondrial ultrastructure. Pretreatment with mangiferin (100 mg/kg b.w. i.p.) for 28 days prevented these mitochondrial alterations, oxidation with energy metabolism and restored the TCA cycle enzyme activities to near normal values following ISPH administration. The structural integrity of the heart was protected by mangiferin in ISPH administered rats when compared to the untreated controls. The present findings suggest that the protective effect of mangiferin can be attributed to its reducing effect on oxidative damage and activation of mitochondrial energy metabolism. These results could be useful to study and understand the cellular events involved in this cardioprotective mechanism of mangiferin. Our studies of mangiferin on heart failure may have important implication for future therapeutic approaches involving in the prevention of cardiovascular diseases.
36
Study On Mango Leaf and Mangiferin Effect of mangiferin on radiation-induced micronucleus formation in cultured human peripheral blood lymphocytes
Ganesh Chandra Jagetia , Venkatasubbaiah A shokaumar Venkatesha Department of Radiobiology, Kasturba Medical College, Manipal, India
Irradiation causes a variety of lesions in important biomolecules of the cell through generation of free radicals leading to genomic instability. DNA strand breaks, acentric fragments, or defective kinetochores are manifested as micronuclei after the first cell division. Chemicals that can trap free radicals may reduce the deleterious effects of ionizing radiation. Mangiferin (MGN), a glucosylxanthone derived from Mangifera indica (mango), was investigated for its ability to reduce the frequency of radiation-induced micronucleated binucleate cells (MNBNCs) in cultured human peripheral blood lymphocytes (HPBLs). HPBL cultures were pretreated with 0, 5, 10, 20, 50, and 100 g/ml of MGN for 30 min before exposure to 3 Gy of (60) Co gamma-radiation. The maximum decline in radiation-induced micronuclei was observed at a concentration of 50 g/ml MGN; thereafter, a nonsignificant elevation in MNBNC frequency was observed at 100 g/ml MGN. Since the lowest MNBNC frequency was observed for 50 g/ml MGN, dose-response studies were undertaken using this concentration. Irradiation of HPBLs with 0, 1, 2, 3, or 4 Gy of gamma-radiation caused a dose-dependent elevation in the MNBNC frequency, while treatment of HPBLs with 50 g/ml MGN 30 min before radiation resulted in significant declines in these frequencies. MGN alone did not alter the proliferation index. Irradiation caused a dose-dependent decline in the proliferation index, while treatment of HPBLs with 50 micro/ml MGN significantly elevated the proliferation index in irradiated cells. MGN treatment reduced hydrogen peroxide-induced lipid peroxidation in HPBLs in a concentration-dependent fashion. In cell-free studies, MGN inhibited the induction of OH (hydroxyl), O2- (superoxide), DPPH (1,1-diphenyl-2-picrylhydrazyl), and ABTS+ (2,-2-azino-bis-3-ethyl benzothiazoline-6-sulphonic acid) radicals in a dose-dependent manner. The results of this study indicate that MGN possesses radioprotective properties by suppressing the effects of free radicals.
37
Study On Mango Leaf and Mangiferin Effect of mangiferin on the development of periodontal disease: involvement of lipoxin A4, anti-chemotaxic action in leukocyte rolling
Roney Rick Carvalho1 , Claudia Helena Pellizzon2, Luis Justulin Jr.2, Sergio Luis Felisbino2 , Wagner Vilegas3, Fernanda Bruni2, Monica Lopes-Ferreira4 , Cllia Akiko Hiruma-Lim1*
1 2
Department of Physiology, Biosciences Institute, So Paulo State University (UNESP), So Paulo, Brazil Department of Morphology, Biosciences Institute, So Paulo State University (UNESP), Botucatu,
SoPaulo, Brazil
3 4
Institute of Chemistry, So Paulo State University (UNESP), Araraquara, SoPaulo, Brazil Special Laboratory of Applied Toxinology (CAT/CEPID) , SoPaulo, Brazil
Mangiferin is a polyphenol compound obtained from mango and has been reported to possess antioxidant and anti-inflammatory properties. We propose to evaluate the influence of mangiferin in preventing and treating experimental periodontitis induced in Wistar rats. Periodontitis was induced in rats by applying a ligature around the lower right first molar. After ligature, groups of animals were submitted orally to the following treatments: saline 10 mL/kg, piroxicam 20 mg/kg or mangiferin 100 mg/kg. On days 1, 4 or 7 after ligature application the alveolar bone loss (ABL) was determined. We evaluated the effect of mangiferin on ABL by histological techniques (alveolar bone loss and cellularity), enzyme immunoassay (lipoxin A4), intravital microscopy (rolling leukocytes and endothelial-leukocyte adhesion), zymographic analyses (metalloproteinases, MMPs 2 and 9), immunohistochemistry (PCNA, COX-2 and CXCR4) and toxicology. of cellularity in mangiferin-treated rats. Treatment with mangiferin inhibited COX-2 expression and the rolling and adhesion of leukocytes, while maintaining normal lipoxin A4 levels. The mangiferin did not interfere in the activity of MMP-2 or -9. The mangiferin-treated rats presented an earlier peak of cell proliferation and augmented angiogenesis in the injured region. Our results have demonstrated promising therapeutic potential of mangiferin both in the prevention and treatment of periodontitis.
38
Study On Mango Leaf and Mangiferin Effect of species variation and processing on phenolic composition and in vitro antioxidant activity of aqueous extracts of Cyclopia spp. (Honeybush Tea)
Elizabeth Joubert1,2* , E. Sian Richards2, 3, J. Debora Merwe2 , Dalene De Beer1, Marena Manley 2, Wentzel C. A. Gelderblom 4
1 2 3 4
ARC Infruitec-Nietvoorbij, Stellenbosch, South Africa. Departments of Food Science and Biochemistry, Stellenbosch University, Stellenbosch , South Africa South Africa Nampak Research & Development , Howard Place,South Africa PROMEC Unit, Medical Research Council, Tygerberg, South Africa
The in vitro antioxidant activity of aqueous extracts prepared from four Cyclopia spp. (unfermented and fermented) was assessed using radical (ABTS+) scavenging, ferric ion reduction, and inhibition of Fe2+-induced microsomal lipid peroxidation as criteria. Aqueous extracts of unfermented and fermented Aspalathus linearis (rooibos) and Camellia sinensis teas (green, oolong, and black) were included as reference samples. Qualitative and quantitative differences in phenolic composition were demonstrated for the Cyclopia spp. The xanthone glycoside, a.k.a. mangiferin, was the major monomeric polyphenol present in the Cyclopia extracts, with both unfermented and fermented C. genistoides extracts containing the highest quantities. Fermentation resulted in a significant reduction in extract yields and their total polyphenolic and individual polyphenol contents. Unfermented plant material should preferentially be used for preparation of extracts, as fermentation significantly ( P < 0.05) lowered antioxidant activity of all species, except in the case of C. genistoides, where the ability to inhibit lipid peroxidation was not affected. Unfermented plant material also retained the highest concentration of mangiferin. Overall, extracts of unfermented Cyclopia were either of similar or lower antioxidant activity as compared to the other teas. However, the presence of high levels of mangiferin merits the use of Cyclopia spp. and, in particular, C. genistoides, as an alternative herbal tea and potential dietary supplement.
39
Study On Mango Leaf and Mangiferin Effects of a natural extract from Mangifera indica L, and its active compound, mangiferin, on energy state and lipid peroxidation of red blood cells
Janet Rodrguez2, Donato Di Pierro1, Magda Gioia1, Susanna Monaco1, Ren Delgado2, Massimiliano Coletta1, Stefano Marini1*
1 2
Department of Exp. Med. And Biochem. Sciences, University of Rome Tor Vergata, Rome, Italy Department of Biomedical Research, Center of Pharmaceutical Chemistry, Havana, Cuba.
Following oxidative stress, modifications of several biologically important macromolecules have been demonstrated. In this study we investigated the effect of a natural extract from Mangifera indica L (Vimang), its main ingredient mangiferin and epigallocatechin gallate (EGCG) on energy metabolism, energy state and malondialdehyde (MDA) production in a red blood cell system. Analysis of MDA, high energy phosphates and ascorbate was carried out by high performance liquid chromatography (HPLC). Under the experimental conditions, concentrations of MDA and ATP catabolites were affected in a dose-dependent way by H2O2. Incubation with Vimang (0.1, 1, 10, 50 and 100 mg/mL), mangiferin (1, 10, 100 mg/mL) and EGCG (0.01, 0.1, 1, 10 mM) significantly enhances erythrocyte resistance to H2O2-induced reactive oxygen species production. In particular, we demonstrate the protective activity of these compounds on ATP, GTP and total nucleotides (NT) depletion after H2O2-induced damage and a reduction of NAD and ADP, which both increase because of the energy consumption following H2O2 addition. Energy charge potential, decreased in H2O2-treated erythrocytes, was also restored in a dose-dependent way by these substances. Their protective effects might be related to the strong free radical scavenging ability described for polyphenols.
40
Study On Mango Leaf and Mangiferin Effects of the mango components mangiferin and quercetin and the putative mangiferin metabolite norathyriol on the transactivation of peroxisome proliferator-activated receptor isoforms
Ashley S. Wilkinson1, Gregory R. Monteith 1, P. Nicholas Shaw1, Chun-Nam Lin2, Michael J. Gidley, Sarah J1,2. Roberts-Thomson 1*
1
School of Pharmacy and Centre for Nutrition and Food Sciences, University of Queensland, Brisbane,
Australia.
2
Graduate Institute of Pharmaceutical Sciences, Kaohsiung Medical University, Taiwan
Mangos are a source of bioactive compounds with potential health-promoting activity. This study evaluated the abilities of the mango components quercetin and mangiferin and the aglycone derivative of mangiferin, norathyriol, to modulate the transactivation of peroxisome proliferator-activated receptor isoforms (PPARs). PPARs are transcription factors important in many human diseases. Through the use of a gene reporter assay it was shown that quercetin inhibited the activation of all three isoforms of PPARs (PPAR IC50=56.3 M; PPAR IC50=59.6 M; PPAR IC50=76.9 M) as did norathyriol (PPAR IC50= 153.5 M; PPAR IC50 = 92.8 M; PPAR IC50=102.4 M), whereas mangiferin did not inhibit the transactivation of any isoform. These findings suggest that mango components and metabolites may alter transcription and could contribute to positive health benefits via this or similar mechanisms.
41
Study On Mango Leaf and Mangiferin Efficacy of mangiferin against Cryptosporidium parvum in a neonatal mouse model
S. Perrucci , G. Fichi , C. Buggiani , G. Rossi , G. Flamini Dipartimento di Patologia Animale, Profilassi ed Igiene degli Alimenti, University of Pisa, Pisa, Italy
The inhibitory activity of mangiferin (50 mg/kg/die and 100 mg/kg/die) on Cryptosporidium parvum was evaluated in a neonatal mouse model and its activity was compared with that of paromomycin (100 mg/kg/die). At 4 days of age, neonatal Swiss conventional outbred mice were experimentally infected by oral administration of 10 cocysts/animal of C. parvum and treated orally for 10 consecutive days, starting 7 days after the experimental infection. One group of mice was left untreated. To evaluate the efficacy of mangiferin, from euthanised mice, 3-mum-thick tissue sections of the intestine were stained with haematoxylin-eosin and periodic acid Schiff. Immunohistochemistry was also used by employing a monoclonal anti-C. parvum antibody. Oocysts were counted and results were expressed as mean oocysts number/intestine. Results obtained show that mangiferin at 100 mg/kg/die has a significant anticryptosporidial activity and that its activity is similar to that showed by the same dose (100 mg/kg/die) of paromomycin. However, both mangiferin and paromomycin were not able to completely inhibit intestinal colonization of C. parvum but only to reduce it. This reduction was calculated at over 80% for both mangiferin and paromomycin with respect to the untreated control. A significant activity was found also for mangiferin at 50 mg/kg/die only after the end of treatment.
42
Study On Mango Leaf and Mangiferin Evaluation of the genotoxic potential of Mangifera indica L. extract (Vimang), a new natural product with antioxidant activity
I. Rodeiro1*, L. Cancino 2, J. E. Gonzlez3 , J. Morffi 1, G. Garrido1 , R. M. Gonzlez1, A. Nuez1 , R . Delgado1
1
Laboratory of Pharmacology, Department of Biomedical Investigations, Center of Pharmaceutical
Chemistry, Havana, Cuba.

2 3
Laboratory of Genotoxicology, Center of Biomedical Investigations (CIBIOMED), Havana, Cuba Laboratory of Genotoxicology, Faculty of Pharmacy, University of Havana, LaCoronela, Havana, Cuba
Mangifera indica L. extract (Vimang) consists of a defined mixture of components (polyphenols, terpenoids, steroids, fatty acids and microelements). It contains a variety of polyphenols, phenolic esters, flavan-3-ols and a xanthone (mangiferin), as main component. This extract has antioxidant action, antitumor and immunemodulatory effects proved in experimental models in both in vitro and in vivo assays. The present study was performed to investigate the genotoxicity potential activity of Vimang assessed through different tests: Ames, Comet and micronucleus assays. Positive and negative controls were included in each experimental series. Histidine requiring mutants of Salmonella typhimurium TA1535, TA1537, TA1538, TA98, TA100 and TA102 strains for point-mutation tests and in vitro micronucleus assay in primary human lymphocytes with and without metabolic activation were performed. In addition, genotoxic effects were evaluated on blood peripheral lymphocytes of NMRI mice of both sexes, which were treated during 2 days with intraperitoneal doses of M. indica L. extract (50-150 mg/kg). The observed results permitted to affirm that Vimang (200-5,000 g/plate) did not increase the frequency of reverse mutations in the Ames test in presence or not of metabolic activation. Results of Comet assay showed that the extract did not induce single strand breaks or alkali-labile sites on blood peripheral lymphocytes of treated animals compared with controls. On the other hand, the results of the micronucleus studies (in vitro and in vivo) showed Vimang induces cytotoxic activity, determined as cell viability or PCE/NCE ratio, but neither increased the frequency of micronucleated binucleate cells in culture of human lymphocytes nor in mice bone marrow cells under our experimental conditions. The positive control chemicals included in each experiment induced the expected changes. The present results indicate that M. indica L. extract showed evidences of light cytotoxic activity but did not induce a mutagenic or genotoxic effects in the battery of assays used.
43
Study On Mango Leaf and Mangiferin Examination of the inhibitory effect of norathyriol in
formylmethionyl-leucyl-phenylalanine-induced respiratory burst in rat neutrophils

Mel-Feng Hsu1*, Shue-Ling Raung 3, Lo-Ti Tsao3, Chun-Nan Lin2 , Jih-Pyang Wang3
1 2 3
Department of Biochemistry, China Medical College, Taichung, Taiwan, Republic of China. School of Pharmacy, Kaohsiung Medical College, Kaohsiung, Taiwan Department of Medica lResearch, Taichung Veterans General Hospital, Taichung, Taiwan, P.R.China
Norathyriol, aglycone of a xanthone C-glycoside mangiferin isolated from Tripterospermum lanceolatum, concentration dependently inhibited the formylmethionyl-leucyl-phenylalanine (fMLP)induced superoxide anion (O2-) generation and O2 consumption in rat neutrophils. In cell-free oxygen radical generating system, norathyriol inhibited the O2- generation during dihydroxyfumaric acid (DHF) autoxidation and in hypoxanthine-xanthine oxidase system. fMLP-induced transient elevation of [Ca2+]i and the formation of inositol trisphosphate (IP3) were significantly inhibited by norathyriol (30 microM) (about 30 and 46% inhibition, respectively). Norathyriol concentration dependently suppressed the neutrophil cytosolic phospholipase C (PLC). In contrast with the marked attenuation of fMLP-induced protein tyrosine phosphorylation (about 70% inhibition at 10 microM norathyriol), norathyriol only slightly modulated the phospholipase D (PLD) activity as determined by the formation of phosphatidic acid (PA) and, in the presence of ethanol, phosphatidylethanol (PEt). Norathyriol did not modulate the intracellular cyclic AMP level. In the presence of NADPH, the phorbol 12-myristate 13-acetate (PMA)-activated particulate NADPH oxidase activity was suppressed by norathyriol in a concentration-dependent manner and the inhibition was noncompetitive with respect to NADPH. Norathyriol inhibited the iodonitrotetrazolium violet (INT) reduction in arachidonic acid (AA)-activated cell-free NADPH oxidase system at the same concentration range as those used in the suppression of PMA-activated particulate NADPH oxidase activity. Taken together, these results suggest that the scavenging ability of norathyriol contributes to the reduction of generated O2-, however, the inhibition of O2- generation from neutrophils by norathyriol is attributed to the blockade of PLC pathway, the attenuation of protein tyrosine phosphorylation, and to the suppression of NADPH oxidase through the interruption of electrons transport.
44
Study On Mango Leaf and Mangiferin Expression profiles of genes involved in the mouse nuclear factor-kappa B signal transduction pathway are modulated by mangiferin
Jos Leiro1* , Juan A. Arranz1, Matilde Yez2, Florencio M. Ubeira1, Manuel L. Sanmartn1, Francisco Orallo2
1
Laboratorio de Parasitologa, Instituto de Investigacin y Anlisis Alimentarios, Universidad de Santiago de Compostela, C/ Constantino Candeira s/n, La Corua, Spain.
Departamento de Farmacologia, Facultad de Farmacia, Universidad de Santiago de Compostela, Santiago de Compostela, Spain
The polyphenol mangiferin (MA) has been shown to have various effects on macrophage function, including inhibition of phagocytic activity and of free radical production. To further characterize the immunomodulatory activity of MA, this study investigated its effects on expression by activated mouse macrophages of diverse genes related to the NF-B signaling pathway, using a DNA hybridization array containing 96 NF-B-related genes and on cytokine levels using a cytokine protein array. MA at 10 M significantly inhibited the expression of (a) two genes of the Rel/NF-B/IB family, RelA and RelB (=I-rel), indicating an inhibitory effect on NF-B-mediated signal transduction; (b)TNF receptor-associated factor 6 (Traf6), indicating probable blockage of activation of the NF-B pathway by lipopolysaccharide (LPS), tumor necrosis factor (TNF), and interleukin 1 (IL-1); (c) other proteins involved in responses to TNF and in apoptotic pathways triggered by DNA damage, including the TNF receptor (TNF-R), the TNF-receptor-associated death domain (TRADD), and the receptor interacting protein (RIP); (d) the extracellular ligand IL-1, again indicating likely interference with responses to IL-1; (e) the pro-inflammatory cytokines IL-1, IL-6, IL-12, TNF- and RANTES (CCL5), and cytokines produced by monocytes and macrophages, including granulocyte colony-stimulating factor (G-CSF),
granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF); (f) other toll-like receptor proteins (in addition to Traf6), including JNK1, JNK2 and Tab1; (g) Scya2 (small inducible cytokine A2=monocyte chemoattractant protein 1); and (h) various intracellular adhesion molecules (ICAMs), and the vascular cell adhesion molecule VCAM-1, which is locally increased in atheromas. The inhibition of JNK1, together with stimulation of c-JUN (i.e. the Jun oncogene) and the previously reported superoxide-scavenging activity of MA, suggests that MA may protect cells against oxidative damage and mutagenesis. Taken together, these results indicate that MA modulates the expression of a large number of genes that are critical for the regulation of apoptosis, viral replication, tumorogenesis, inflammation and various autoimmune diseases, and raise the possibility that it may be of value in the treatment of inflammatory diseases and/or cancer.
45
Study On Mango Leaf and Mangiferin Fe (III) improves antioxidant and cytoprotecting activities of mangiferin
Gilberto L. Pardo-Andreu1,2 *, Carlos Snchez-Baldoqun1 , Rizette Avila-Gonzlez 1, Ren Delgado 1, Zeki Naal 2, Carlos Curti2
1
Departamento de Investigaciones Biomdicas, Centro de Qumica Farmacutica, Calle 200, Esq. 21,Playa, Ciudad de La Habana, Cuba.
Departamento de Fiasicae Quiamica, Faculdade de Cieoncias Farmaceouticas deRibeirao Preto, Universidade de Sao Paulo, Ribeirao Preto, Razil
Iron-induced oxidative stress has been implicated in several pathological processes. In the present work we provide evidence for the formation of a mangiferin:Fe (III) complex (2:1), shown by means of either iron-induced changes in the UV/visible spectrum of mangiferin or by reduction of the anodic current peak in the voltammogram of that compound; we demonstrate, in addition, that the ferric complex is more effective than mangiferin itself in scavenging superoxide radicals generated by pyrogallol autoxidation, as well as in protecting hepatocytes from reactive oxygen species mediated hypoxia/reoxygenation injury. Moreover, we found that the mangiferin:Fe (III) complex reacts more readily with horseradish peroxidase/H2O2 than does mangiferin by itself. We postulate that mangiferin could afford protection against iron/reactive oxygen species-mediated pathological processes by means of both iron chelating and iron-stimulated superoxide radical scavenging activity.
Immunomodulatory activity of alcoholic extract of Mangifera indica L. in mice

Neelam Makare, Subhash Bodhankar *, Viond Rangari Poona College of Pharmacy, Bharati Vidyapeeth (Deemed University), Pune, India.
Mangifera indica Linn, a plant widely used in the traditional medicinal systems of India, has been reported to possess antiviral, antibacterial and anti-inflammatory activities. In the present study, the alcoholic extract of stem bark of Mangifera indica Linn (Extract I containing mangiferin 2.6%), has been investigated for its effect on cell mediated and humoral components of the immune system in mice. Administration of test extract I produced increase in humoral antibody (HA) titre and delayed type hypersensitivity (DTH) in mice. It is concluded that test extract I is a promising drug with immunostimulant properties.
46
Study On Mango Leaf and Mangiferin Gastroprotective effect of mangiferin, a xanthonoid from Mangifera indica, against gastric injury induced by ethanol and indomethacin in rodents
Ana Carla s. Carvalho1 , Marjorie M. Guedes1 , Antonia L. de Souza2 , Maria T.s. Trevisan 2, Alana F. Lima 1, lvia A. Santos1, Vletla S. Rao1
1 2
Department of Physiology and Pharmacology, Federal University of Cear, Fortaleza, CE, Brazil. Department of Organic and Inorganic Chemistry, Federal University of Cear, Fortaleza, CE, Brazil.
In search of novel gastroprotective agents, mangiferin, a naturally occurring glucosylxanthone from Mangifera indica L. (Anacardiaceae), was evaluated in mice on gastric injury induced by ethanol and indomethacin. The effects of mangiferin on gastric mucosal damage were assessed by determination of changes in mean gastric lesion area or ulcer score in mice and on gastric secretory volume and total acidity in 4-h pylorus-ligated rats. Mangiferin (3, 10 and 30 mg/kg, P. O.) significantly attenuated the gastric damage induced by ethanol by 30, 35, and 63 %, and of indomethacin by 22, 23 and 57 %, respectively. N-Acetylcysteine (750 mg/kg, I. P.) and lansoprazole (30 mg/kg, P. O.) used as positive controls in these ulcerogenic models resulted in 50 % and 76 % suppression of gastric injury, respectively. In 4-h pylorus-ligated rats, intraduodenally applied mangiferin (30 mg/kg) caused significant diminutions in gastric secretory volume and total acidity. In addition, like N-acetylcysteine, a donor of sulfhydryls, mangiferin effectively prevented the ethanol-associated depletion of gastric mucosal non-protein sulfhydryl content in mice, suggesting an antioxidant action. These findings provide evidence that mangiferin affords gastroprotection against gastric injury induced by ethanol and indomethacin most possibly through the antisecretory and antioxidant mechanisms of action.
47
Study On Mango Leaf and Mangiferin In vitro effects of Mangifera indica and polyphenols derived on ABCB1/P-glycoprotein activity
Elisabetta Chieli1 *, Nadia Romiti1 , Idania Rodeiro2 , Gabino Garrido3
1
Dipartimento di Patologia Sperimentale, Facolt di Medicina e Chirurgia, Universit degli Studi di Pisa,
Pisa, Italy
2 3
Laboratorio de Farmacologa, Centrode Qumica Farmacutica, Habana, Cuba Departamento de Qumica y Farmacia, Facultad de Ciencias, Universidad Catlica del Norte, Antofagasta,
Chile
Many plant-derived compounds, including polyphenols, are able to affect the function of MDR-1/P-glycoprotein (P-gp ABCB1) multidrug transporter, leading to potential herb-drug interactions. This study evaluated the effects of mango (Mangifera indica L.) stem bark extract, MSBE, and related phenols on P-gp activity in both the HK-2 proximal tubule cell line, constitutively expressing P-gp, and in a Caco-2 cell sub-line selected by resistance to vincristine (Caco-2/VCR) and overexpressing P-gp. The effects of MSBE, mangiferin, norathyriol, catechin, quercetin and gallic acid on P-gp activity were tested by the rhodamine-123 accumulation as well as by the Calcein-AM assays. Effects on esterase activity, which could influence the results of Calcein-AM test, were also assessed. All investigated compounds except for catechin and gallic acid inhibited P-gp activity in HK-2 cells, in the order of mangiferin<norathyriol<quercetin<MSBE. MSBE, quercetin and norathyriol also inhibited significantly esterase activity. Similar effects were obtained in resistant Caco-2/VCR cells, but were negligible in the wild-type ones, expressing low amounts of P-gp. Our results demonstrate, for the first time, that M. indica and polyphenols derived may affect the activity of the multidrug transporter P-gp ABCB1, suggesting the possibility of herb-drug interactions to be explored in depth.
48
Study On Mango Leaf and Mangiferin Immunotherapeutic effects of mangiferin mediated by the inhibition of oxidative stress to activated lymphocytes, neutrophils and macrophages
S. Muruganandan , J. Lal , P.K. Gupta Division of Pharmacology and Toxicology, Indian Veterinary Research Institute, Izatnagar, India
The effect of mangiferin, a naturally occurring xanthone glucoside on cyclophosphamide-induced immunotoxicity and its mode of action in the immune system were investigated. To induce immunotoxicity, adult male Wistar rats were injected weekly with cyclophosphamide intraperitoneally at 100 mg/kg bodyweight. Mangiferin was injected intraperitoneally at 10 and 20 mg/kg daily for 14 days. Levamisole (3 mg/kg, i.p., daily for 14 days), a known immunostimulant that acts in immunosuppressive conditions was used as a standard drug. The effect of mangiferin on the primary immune response to ovalbumin (200 g/rat, s.c.) was assessed at weekly intervals by measuring the serum ovalbumin-specific IgM levels. The organ weights and cellularity of spleen, thymus and bone marrow, haematology, T and B cell-dependent mitogen stimulation of splenocytes were assessed for the cellular response. Oxidative changes in lymphocytes, neutrophils and macrophages were measured at the end of the study. As well, the in vitro effect of mangiferin on cytotoxicity caused by H2O2 in primary lymphocytes was studied. The decrease in the lymphoid organ weights, cellular responses and antigen-specific IgM levels by cyclophosphamide treatment were significantly increased by repeated intraperitoneal administration of mangiferin. The enhanced lipid peroxidation and decreased catalase and superoxide dismutase activities found in lymphocytes, polymorphonuclear cells (PMN) and macrophages from cyclophosphamide treated rats were significantly ameliorated in mangiferin treated groups. The tissue injury caused by cyclophosphamide treatment was significantly suppressed by mangiferin as shown by the decrease in serum creatine phosphokinase (CPK) activity. In vitro experiments showed that pretreatment of lymphocytes with mangiferin protected from the toxicity induced by H2O2, further confirming the in vivo findings. From this study, it is evident that mangiferin exhibits an immunoprotective role mediated through the inhibition of reactive intermediate-induced oxidative stress in lymphocytes, neutrophils and macrophages.
49
Study On Mango Leaf and Mangiferin In vitro effects of mangiferin on superoxide concentrations and expression of the inducible nitric oxide synthase, tumournecrosis factor- and transforming growth factor- genes
Jos Manuel Leiro1* ,Ezequiel Alvarez2 , Juan Alberto Arranz 1, Isabel Gonzlez Siso3, Francisco Orallo2
1
Departamento de Microbiologa y Parasitologa, Universidad de Santiago de Compostela, Santiago de
Compostela, Spain
2
Departamento de Farmacologa, Facultad de Farmacia, Universidad de Santiago de Compostela,
Santiagode Compostela, Spain

3
Laboratorio de Bioqumica, Departamento de Biologa Celulary Molecular, Facultad de Ciencias,
Universidad de LaCoruna, Coruna, Spain
This study investigated the effects of the natura lpolyphenol mangiferin (MA) on superoxide anion (O2-) production, xanthine oxidase (XO) activity, vascular contractility, inducible nitric oxide synthase (iNOS) mRNA levels, tumour necrosis factor-alpha (TNF-) mRNA levels, and tumour growth factor-beta (TGF-) mRNA levels. O2- was generated by the hypoxanthine-xanthineoxidase (HX-XO) and phenazine methosulphate (PMS)-NADH systems. XO activity was determined by measurement of uric acid production with xanthine as substrate. Vascular contraction experiments were performed with intactrat aortic rings. iNOS, TNF- and TGF- gene expression in rat macrophages stimulated in vivo with 3% thioglycollate and invitro with 100 ng/mL lipopolysaccharide and 10 U/mL of interferon-gamma were evaluated semiquantitatively by the retrotranscriptase-polymerase chain reaction. MAat10-1 M, like the known O2- scavenger superoxide dismutase (1U/mL), scavenged O2- produced by the HX/XO and PMS-NADHs ystems. By contrast MAat1-1 mM, unlike allopurinol (10mM), was unable to inhibit XO activity. MA at 1-10 mM did not modify resting tone or the contractile responsese licited by 1 M phenylephrine or 1 mM phorbol 12-myristate13-acetateinrataorta. MA at 1-10 mM, like dexamethasone (100mM), decreased iNOS mRNA levels inactivated macrophages. At 100 mM, MA also reduced TNF- mRN Alevels, but increased TGF- mRNA levels. These results thus indicate that MA is an O2- scavenger and that itinhibits expression of the iNOS and TNF- genes, suggesting that it may be of potential value in the treatment of inflammatory and/or neurodegenerative disorders. Inaddition, the finding that MA enhances TGF- gene expression suggests that this polyphenol might also be of value in the prevention of cancer, autoimmune disorders, atherosclerosis and coronary heart disease.
50
Study On Mango Leaf and Mangiferin In vitroeffects of the polyphenols on the resveratrol, scuticociliate mangiferin fish and
(-)-epigallocatechin-3-gallate Philasterides dicentrarchi
pathogen
J. Leiro*, J. A. Arranz, A. Param, M. F. Alvarez, M. L. Sanmartn Laboratorio de Parasitologa, Instituto de Investigacin y Anlisis Alimentarios, Universidad de Santiago de Compostel, Santiago de Compostela, Spain
This study investigated the in vitro effects of the polyphenols resveratrol, mangiferin and (-)-epigallocatechin-3-gallate (EGCG) on the histiophagous ciliatePhilasterides dicentrarchi, which causes fatal scuticociliatosis in farmed turbot Scophthalmus maximus L. of the 3 polyphenols, resveratrol showed strongest antiprotozoal activity, reducing ciliate density after 1 wk culture by, on average, 91% at 50 PAM, and 96% at 500 PAM. EGCG reduced ciliate density by, on average, 93% 500 PAM, with no significant effect at 50 PAM. Mangiferin reduced ciliate density by, on average, 5 at 500 PAM, again with no significant effect at 50 PAM. In view of these findings, we discuss the potetial utility of chemotherapy with polyphenols as a strategy for the control of scuticociliatosis in farmed turbot.
Mangiferin,a glucosylxanthone,protects against the radiation-induced micronuclei formation in the cultured human peripheral blood lymphocytes
Ganesh Chandra Jagetia, V.A. Venkatesha Department of Radiobiology, Kasturba Medical College, Manipal, India
The effect of mangiferin (MGN), a glucosylxanthone, derived from Mangifera indica was studied on the radiation-induced DNA damage in the cultured human peripheral blood lymphocytes (HPBLs) by micronucleus assay, where HPBLs were treated with 0, 5, 10, 20, 50, 100 g/ml of mangiferin 30 min before exposure to 3 Gy of
60
Co -radiation. Treatment of HPBLs with 50 g/ml reduced the
radiation-induced micronuclei to the maximum extent. Irradiation of HPBLs to 0, 1, 2, 3, or 4 Gy resulted in a dose-dependent increase in the frequency of micronuclei, while treatment of HPBLs with 50 g/ml MGN before exposure to 0, 1, 2, 3, or 4 Gy of
60
Co -radiation resulted in a significant decline in the
frequency of micronuclei when compared with the untreated irradiation group.
51
Study On Mango Leaf and Mangiferin In vivo and in vitro anti-inflammatory activity of Mangifera indica L. extract (VIMANG)
Gabino Garrido 1*, Deyarina Gonzlez1 , Yeny Lemus 1, Dagmar Garca 1, Lizt Lodeiro 1, Gypsy Quintero 1, Carla Delporte2, Alberto J. Nez-Sells 1, Ren Delgado1
1 2
Laboratorio de Farmacologa. Centro de Qumica Farmacutica, Atabey, Playa, Habana, Cuba Facultad de Ciencias Qumicasy Farmacuticas, Universidad de Chile, Santiago, Chile
A standard aqueous extract of Mangifera indica L., used in Cuba as an antioxidant under the brand name of VIMANG, was tested in vivo for its anti-inflammatory activity using commonly accepted assays. M. indica extract, administered topically (0.5-2 mg/ear), reduced ear edema induced by arachidonic acid (AA) and phorbol myristate acetate (PMA, ED50 = 1.1 mg per ear) in mice. In the PMA model, M. indica extract also reduced myeloperoxidase (MPO) activity. This extract p.o. administered also inhibited tumor necrosis factor alpha (TNF) serum levels in both models of inflammation (AA, ED50 = 106.1 mgkg-1 and PMA, ED50 = 58.2 mgkg-1). In vitro studies were performed using the macrophage cell line RAW 264.7 stimulated with pro-inflammatory stimuli (LPS-IFN or the calcium ionophore A23187) to determine PGE2 or LTB4 release, respectively. The extract inhibited the induction of PGE2 with IC50 = 64.1 gml-1 and LTB4 IC50 = 22.9 gml-1. M. indica extract also inhibited human synovial secretory phospholipase (PL)A2 with IC
50
= 0.7 gml-1. These results represent an important contribution to the elucidation of the
mechanism involved in the anti-inflammatory and anti-nociceptive effects reported by the standard M. indica extract VIMANG.
52
Insulin secretion is stimulated by ethanol extract of anemarrhena asphodeloides in isolated islet of healthy wistar and diabetic Goto-Kakizaki Rats
Hoa NK, Phan DV, Thuan ND, Ostenson CG Department of Pharmacology, Hanoi Medical University, Hanoi, Vietnam
The hypoglycemic effect of extract of Anemarrhena asphodeloides has been accounted for by the substance mangiferin which increases insulin sensitivity. The present study aimed to investigate whether an ethanol extract of Anemarrhena asphodeloides would stimulate insulin secretion and if so, further elucidate the mechanism behind this effect. Isolated pancreatic islets of normal Wistar rats and spontaneously diabetic Goto-Kakizaki (GK) rats were batch incubated or perifused to study effect of Anemarrhena asphodeloides extract (TH2) on insulin release. At 3.3 mM glucose, 2, 4, and 8 mg/ml TH2 increased the insulin release of Wistar rat islets 2.5-, 4.1-, and 5.7-fold, respectively (p < 0.05) and of GK rat islets 1.7-, 3.0-, and 6.3-fold, respectively (p < 0.01). Similarly at 16.7 mM glucose, 2, 4 and 8 mg/ml TH2 increased insulin release of Wistar rat islets 1.5-, 2.2-, and 3.8-fold, respectively (p < 0.05) and of GK rat 2.5-, 4.2-, and 11.9-fold, respectively (p < 0.01). In perifusions of islets, TH2 also increased insulin secretion that returned to basal levels when TH2 was omitted from the perifusate. Mangiferin had no effect on insulin secretion of islets. In islets depolarized by 30 mM KCl and B-cell K-ATP channels kept open by 0.25 mM diazoxide, TH2 (8 mg/ml) further enhanced insulin secretion at 3.3 but not at 16.7 mM glucose. Pertussis toxin suppressed the insulin stimulating effect of 2 and 8 mg/ml TH2 by 35 % and 47 % (p < 0.05 and p < 0.001, respectively). Ethanol extract of the roots of Anemarrhena asphodeloides contains a substance, TH2, that stimulates insulin secretion both at 3.3 and 16.7 mM glucose in islets of normal Wistar and diabetic GK rats. The mechanism behind TH2-stimulated insulin secretion involves an effect on the exocytotic machinery of the B-cell, mediated via pertussis toxin-sensitive Gi- (or Ge-) proteins.
53
Interaction of Vimang (Mangifera indica L. extract) with Fe(III) improves its antioxidant and cytoprotecting activity
Pardo-Andreu GL, Snchez-Baldoqun C, Avila-Gonzlez R, Yamamoto ET, Revilla A, Uyemura SA, Naal Z, Delgado R, Curti C Departamento de Investigaciones Biomdicas, Centro de Qumica Farmacutica, Habana, Cuba
A standard aqueous stem bark extract from selected species of Mangifera indica L. (Anacardiaceae)Vimang, whose major polyphenolic component is mangiferin, displays potent in vitro and in vivo antioxidant activity. The present study provides evidence that the Vimang-Fe(III) mixture is more effective at scavenging 2,2-diphenyl-1-picrylhydrazyl (DPPH) and superoxide radicals, as well as in protecting against t-butyl hydroperoxide-induced mitochondrial lipid peroxidation and hypoxia/reoxygenationinduced hepatocytes injury, compared to Vimang alone. Voltammetric assays demonstrated that Vimang, in line with the high mangiferin content of the extract, behaves electrochemically like mangiferin, as well as interacts with Fe(III) in close similarity with mangiferin's interaction with the cation. These results justify the high efficiency of Vimang as an agent protecting from iron-induced oxidative damage. We propose Vimang as a potential therapy against the deleterious action of reactive oxygen species generated during iron-overload, such as that occurring in diseases like beta-thalassemia, Friedreich's ataxia and haemochromatosis.
54
Study On Mango Leaf and Mangiferin Iron complexing activity of mangiferin,a naturally occurring
glucosylxanthone,inhibits mitochondrial lipid peroxidation induced by Fe2+-citrate.

Andreu GP, Delgado R, Velho JA, Curti C, Vercesi AE Departamento de Patologia Clnica, Faculdade de Cincias Mdicas, Universidade Estadual de Campinas, SP, Brasil
Mangiferin,a naturally occurring glucosylxanthone, has been described as having antidiabetic, antiproliferative, immunomodulatory and antioxidant activities. In this study we report for the first time the iron-complexing ability of mangiferin as a primary mechanism for protection of rat liver mitochondria against Fe2+-citrate induced lipid peroxidation. Thiobarbituric acid reactive substances and antimycin A-insensitive oxygen consumption were used as quantitative measures of lipid peroxidation. Mangiferin at 10 M induced near-full protection against 50 M Fe2+-citrate-induced mitochondrial swelling and loss of mitochondrial transmembrane potential (DeltaPsi). The IC50 value for mangiferin protection against Fe2+-citrate-induced mitochondrial thiobarbituric acid reactive substance formation (9.021.12 M) was around 10 times lower than that for tert-butylhydroperoxide mitochondrial induction of thiobarbituric acid reactive substance formation. The xanthone derivative also inhibited the iron citrate induction of mitochondrial antimycin A-insensitive oxygen consumption, stimulated oxygen consumption due to Fe2+ autoxidation and prevented Fe3+ ascorbate reduction. Absorption spectra of mangiferin-Fe2+/Fe3+ complexes also suggest the formation of a transient charge transfer complex between Fe2+ and mangiferin, accelerating Fe2+ oxidation and the formation of a more stable Fe3+-mangiferin complex unable to participate in Fenton-type reaction and lipid peroxidation propagation phase. In conclusion, these results show that in vitro antioxidant activity of mangiferin is related to its iron-chelating properties and not merely due to the scavenging activity of free radicals. These results are of pharmacological relevance since mangiferin and its naturally contained extracts could be potential candidates for chelation therapy in diseases related to abnormal intracellular iron distribution or iron overload.
55
Study On Mango Leaf and Mangiferin Isolation of a human intestinal bacterium that transforms mangiferin to norathyriol and inducibility of the enzyme that cleaves a C-Glucosyl bond.
Sanugul K, Akao T, Li Y, Kakiuchi N, Nakamura N, Hattori M Institute of Natural Medicine, Toyama Medical and Pharmaceutical University, Toyama, Japan
The C-glucosyl bond of C-glucosides generally tolerates acid and enzymatic hydrolysis. Many C-glucosides are cleaved by human intestinal bacteria. We isolated the specific bacterium involved in the metabolism of mangiferin (2-beta-D-glucopyranosyl-1,3,6,7-tetrahydroxyxanthone), C-glucosyl xanthone, from a mixture of human fecal bacteria. The anaerobic Bacteroides species named MANG, transformed mangiferin to the aglycone, norathyriol, suggesting cleavage of a C-glucosyl bond. However, B. sp. MANG cleaved C-glucosyl in a dose- and time-dependent manner only when cultivated in the presence of mangiferin. Cleavage was abolished by inhibitors of RNA and protein syntheses, such as rifampicin and chloramphenicol, respectively, indicating that the enzyme that cleaves C-glucosyl is induced by mangiferin. In contrast, mangiferin did not affect bacterial - and beta-glucosidase activities under any conditions. The C-glucosyl-cleavage in cell-free extracts was not altered by potent glucosidase inhibitors such as 1-deoxynojirimycin and gluconolactone. Therefore, the C-glucosyl-cleaving enzyme substantially differs from known glucosidases that cleave O-glucosides. This is the first description of a specific intestinal bacterium that is involved in the metabolism of mangiferin and which produces a novel and inducible C-glucosyl-cleaving enzyme.
56
Study On Mango Leaf and Mangiferin Mangifera indica L. extract (Vimang) and its main polyphenol mangiferin prevent mitochondrial oxidative stress in atherosclerosis-prone hypercholesterolemic mouse.
Pardo-Andreu GL, Paim BA, Castilho RF, Velho JA, Delgado R, Vercesi AE, Oliveira HC Departamento de Investigaciones Biomdicas, Centro de Qumica Farmacutica, Habana, Cuba
Atherosclerosis is linked to a number of oxidative events ranging from low-density lipoprotein (LDL) oxidation to the increased production of intracellular reactive oxygen species (ROS). We have recently demonstrated that liver mitochondria isolated from the atherosclerosis-prone hypercholesterolemic LDL receptor knockout (LDLr-/-) mice have lower content of NADP(H)-linked substrates than the controls and, as consequence, higher sensitivity to oxidative stress and mitochondrial membrane permeability transition (MPT). In the present work, we show that oral supplementation with the antioxidants Mangifera indica L. extract (Vimang) or its main polyphenol mangiferin shifted the sensitivity of LDLr-/- liver mitochondria to MPT to control levels. These in vivo treatments with Vimang and mangiferin also significantly reduced ROS generation by both isolated LDLr-/- liver mitochondria and spleen lymphocytes. In addition, these antioxidant treatments prevented mitochondrial NAD(P)H-linked substrates depletion and NADPH spontaneous oxidation. In summary, Vimang and mangiferin spared the endogenous reducing equivalents (NADPH) in LDLr-/- mice mitochondria correcting their lower antioxidant capacity and restoring the organelle redox homeostasis. The effective bioavailability of these compounds makes them suitable antioxidants with potential use in atherosclerosis susceptible conditions.
57
Study On Mango Leaf and Mangiferin Mangifera indica L. extract (Vimang) and mangiferin modulate mouse humoral immune responses.
Garca D, Leiro J, Delgado R, Sanmartn ML, Ubeira FM Laboratorio de Farmacologa, Centro de Qumica Farmacutica, Ciudad de la Habana (Havana City), Cuba
The present study investigated the effects of orally administered Vimang (an aqueous extract of Mangifera indica) and mangiferin (the major polyphenol present in Vimang) on mouse antibody responses induced by inoculation with spores of microsporidian parasites. Inoculation induced specific antibody production with an exponential timecourse, peaking after about one month. Vimang significantly inhibited this antibody production from about three weeks post-inoculation, and most markedly by four weeks post-inoculation; by contrast, mangiferin had no significant effect. Determination of Ig isotypes showed that the IgM to IgG switch began about four weeks post-inoculation, with IgG2a predominating. Vimang significantly inhibited IgG production, but had no effect on IgM. Mangiferin did no affect either IgM or IgG2a, but significantly enhanced production of IgG1 and IgG2b. Neither Vimang nor mangiferin enhanced specific antibody secretion by splenic plasma cells from mice inoculated with microsporidian spores, whether administered in vivo before serum extraction or in vitro to the culture medium. Inoculation with spores induced splenomegaly, which was significantly reduced by Vimang and significantly enhanced by mangiferin. These results suggest that components of Mangifera indica extracts may be of potential value for modulating the humoral response in different immunopathological disorders.
58
Study On Mango Leaf and Mangiferin Mangifera indica L. extract (Vimang) inhibits 2-deoxyribose damage induced by Fe (III) plus ascorbate.
Pardo-Andreu GL, Delgado R, Nez-Sells AJ, Vercesi AE Departamento de Patologia Clnica, Faculdade de Cincias Mdicas, Universidade Estadual de Campinas, Campinas, SP, Brasil
Vimang is an aqueous extract of selected species of Mangifera indica L., used in Cuba as a nutritional antioxidant supplement. Many in vitro and in vivo models of oxidative stress have been used to elucidate the antioxidant mechanisms of this extract.To further characterize the mechanism of Vimang action, its effect on the degradation of 2-deoxyribose induced by Fe (III)-EDTA plus ascorbate or plus hypoxanthine/xanthine oxidase was studied. Vimang was shown to be a potent inhibitor of 2-deoxyribose degradation mediated by Fe (III)-EDTA plus ascorbate or superoxide (O2-). The results revealed that Vimang, at concentrations higher than 50 M mangiferin equivalent, was equally effective in preventing degradation of both 15 mM and 1.5 mM 2-deoxyribose. At a fixed Fe (III) concentration, increasing the concentration of ligands (either EDTA or citrate) caused a significant reduction in the protective effects of Vimang. When ascorbate was replaced by O2- (formed by hypoxanthine and xanthine oxidase) the protective efficiency of Vimang was also inversely related to EDTA concentration. The results strongly indicate that Vimang does not block 2-deoxyribose degradation by simply trapping *OH radicals. Rather, Vimang seems to act as an antioxidant by complexing iron ions, rendering them inactive or poorly active in the Fenton reaction.
59
Study On Mango Leaf and Mangiferin Mangifera indica L. extract (Vimang) inhibits Fe2+-citrate-induced lipoperoxidation in isolated rat liver mitochondria.
Pardo Andreu G, Delgado R, Velho J, Inada NM, Curti C, Vercesi AE Departamento de Patologia Clnica, Faculdade de Cincias Mdicas, Universidade Estadual de Campinas Campinas, SP, Brasil
The extract of Mangifera indica L. (Vimang) is able to prevent iron mediated mitochondrial damage by means of oxidation of reduced transition metals required for the production of superoxide and hydroxyl radicals and direct free radical scavenging activity. In this study we report for the first time the iron-complexing ability of Vimang as a primary mechanism for protection of rat liver mitochondria against Fe2+ -citrate-induced lipoperoxidation. Thiobarbituric acid reactive substances (TBARS) and antimycin A-insensitive oxygen consumption were used as quantitative measures of lipoperoxidation. Vimang at 10 M mangiferin concentration equivalent induced near-full protection against 50 M Fe2+ -citrate-induced mitochondrial swelling and loss of mitochondrial transmembrane potential (DeltaPsi). The IC50 value for Vimang protection against Fe2+ -citrate-induced mitochondrial TBARS formation (7.891.19 M) was around 10 times lower than that for tert-butylhydroperoxide mitochondrial induction of TBARS formation. The extract also inhibited the iron citrate induction of mitochondrial antimycin A-insensitive oxygen consumption, stimulated oxygen consumption due to Fe2+ autoxidation and prevented Fe3+ ascorbate reduction. The extracted polyphenolic compound, mainly mangiferin, could form a complex with Fe2+, accelerating Fe2+ oxidation and the formation of more stable Fe3+ -polyphenol complexes, unable to participate in Fenton-type reactions and lipoperoxidation propagation phase. The strong DPPH radical scavenging activity with an apparent IC50 of 2.450.08 M suggests that besides its iron-complexing capacity, Vimang could also protect mitochondria from Fe2+ -citrate lipoperoxidation through direct free radical scavenging ability, mainly lipoperoxyl and alcoxyl radicals, acting as both a chain-breaking and iron-complexing antioxidant. These results are of pharmacological relevance since Vimang could be a potential candidate for antioxidant therapy in diseases related to abnormal intracellular iron distribution or iron overload.
60
Mangifera indica L. extract attenuates glutamate-induced neurotoxicity on rat cortical neurons.

Lemus-Molina Y, Snchez-Gmez MV, Delgado-Hernndez R, Matute C Laboratorio de Farmacologa, Departamento de Investigaciones Biomdicas, Centro de Qumica Farmacutica, Habana, Cuba
Overstimulation of ionotropic glutamate receptors causes excitotoxic neuronal death contributing to neurodegenerative disorders. Massive influx of calcium in excitotoxicity provokes alterations in the membrane potential of mitochondria and increases the production of reactive oxygen species. Here we report that Mangifera indica L. extracts (MiE) prevent glutamate-induced excitotoxicity in primary cultured neurons of the rat cerebral cortex. To evaluate the effects of MiE on excitotoxicity, cells were stimulated with l-glutamic acid (50M; 10min) alone or in the presence of MiE. Maximal protection (56%) was obtained with 2.5g/mL of MiE. In turn, we measured the effects of MiE on excitotoxic-induced oxidative stress and mitochondrial depolarization by fluorimetry using 5,6-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate and tetramethylrhodamine, respectively. Both parameters were effectively reduced by MiE at concentrations which showed neuroprotection. Mangiferin, an antioxidant polyphenol which is a major component of MiE, was also effective in preventing neuronal death, oxidative stress and mitochondrial depolarization. Maximal protection (64%) was obtained at 12.5g/mL of mangiferin which also attenuated oxidative stress and mitochondrial depolarization at the neuroprotective concentrations. Together, these results indicate that MiE is an efficient neuroprotector of excitotoxic neuronal death, indicates that mangiferin carries a substantial part of the antioxidant and neuroprotective activity of MiE, and that this natural extract has therapeutic potential to treat neurodegenerative disorders.
61
Study On Mango Leaf and Mangiferin Mangiferin ameliorates scopolamine-induced learning deficits in mice
Jung K, Lee B, Han SJ, Ryu JH, Kim DH Department of Life and Nanopharmaceutical Sciences, Kyung Hee University, Seoul, Korea
The aim of this study was to evaluate the effects of Anemarrhena asphodeloides Bunge (AA) on cholinergic memory deficits in mice. This agent has previously been used as an antipyretic, anti-inflammatory, anti-diabetic, and antidepressant in traditional Chinese medicine. Mangiferin was isolated from AA and showed a dose-dependent inhibition of acetylcholinesterase (AChE) activity (IC50 value, 62.8 M). Cholinergic dysfunction was induced in mice by administering scopolamine, and the animals were then tested using the passive avoidance test as well as the Morris water maze test. Mangiferin (20 mg/kg, p.o.) significantly reversed scopolamine-induced deficits in the passive avoidance test, and also improved escape latencies in training trials and increased swimming times in the Morris water maze test (p<0.05). Mangiferin also reduced acetylcholine and tumor necrosis factor (TNF)- levels induced by scopolamine in mice brain (p<0.05) and inhibited nuclear factor (NF)-kappaB activation in scopolamine or TNF--stimulated BV-2 glial cells. These results suggest that mangiferin can improve long-term cholinergic memory deficits by AChE inhibition or cholinergic receptor stimulation and inhibition of NF-kappaB activation.
62
Study On Mango Leaf and Mangiferin Mangiferin inhibits cyclooxygenase-2 expression and prostaglandin E2 production in activated rat glial cells
Bhatia HS, Candelario-Jalil E, de Oliveira AC, Olajide OA, Martnez-Snchez G, Fiebich BL Neurochemistry Research Group, Department of Psychiatry, University of Freiburg Medical School, Freiburg, Germany
Mangiferin, a naturally occurring glucosylxanthone, has potent antioxidant and anti-inflammatory properties, as demonstrated in several reports. However, very limited information is available on the effects of this natural polyphenol on glial activation. Thus, the aim of this study was to examine whether mangiferin is able to reduce prostaglandin E2 (PGE2) and 8-iso-prostaglandin F(2) (8-iso-PGF(2)) production by lipopolysaccharide (LPS)-activated primary rat glia. glial cells were stimulated with 10ng/ml of LPS in the presence or absence of different concentrations of mangiferin (1-50 M). After 24h incubation, culture media were collected to measure the production of PGE2 and 8-iso-PGF(2) using enzyme immunoassays. Protein levels of cyclooxygenase (COX)-1 and COX-2 were studied by immunoblotting after 24h of incubation with LPS. Mangiferin potently reduced LPS-induced PGE2 synthesis and the formation of 8-iso-PGF(2). Interestingly, mangiferin dose-dependently reduced LPS-induced COX-2 protein synthesis without modifying COX-2 transcription. This was due to a decrease in COX-2 transcript stability. However, mangiferin did not modify LPS-mediated phosphorylation of p38 mitogen-activated protein kinase (p38 MAPK), a key factor involved in enhancing COX-2 mRNA stability and COX-2 translation in primary glia. Mangiferin had no effects on LPS-induced expression of inducible nitric oxide synthase (iNOS) or TNF- production. Taken together, results from the present study indicate that mangiferin is able to limit glial activation, in terms of attenuation of PGE2 production, free radical formation and reduction in COX-2 synthesis induced by LPS. These data suggest that modulation of glial activation might contribute to the mechanism of cerebral protection by mangiferin.
63
Study On Mango Leaf and Mangiferin Mangiferin Inhibits Passive Cutaneous Anaphylaxis Reaction and Pruritus in Mice
Lee B, Trung Trinh H, Bae EA, Jung K, Kim DH Department of Life and Nanopharmaceutical Sciences and Department of Pharmaceutical Science, Kyung Hee University Dongdaemun-ku, Seoul, Korea
The antiallergic effect of mangiferin isolated from the rhizome of Anemarrhena asphodeloides Bunge (family Liliaceae) was measured in vitro and in vivo. Orally and intraperitoneally administered
mangiferin potently inhibited passive cutaneous anaphylaxis (PCA) reaction induced by IgE-antigen complex as well as pruritus induced by compound 48/80 in mice. Mangiferin also inhibited the expression of the proinflammatory cytokine TNF- and the IgE-switching cytokine IL-4 as well as transcription factor NF- kappaB activation in RBL-2H3 cells stimulated by IgE-antigen complex. These findings suggest that mangiferin may improve PCA reaction and pruritus.
64
Study On Mango Leaf and Mangiferin Mangiferin protects against 1-methyl-4-phenylpyridinium toxicity mediated by oxidative stress in N2a cells
Amazzal L, Laptre A, Quignon F, Bagrel D Laboratoire d'Ingnierie Molculaire et Biochimie Pharmacologique, Universit Paul Verlaine-Metz, Metz, France
1-Methyl-4-phenyl-pyridine
ion
(MPP+),
the
active
metabolite
of
1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induces a Parkinsonian syndrome in humans and animals, a neurotoxic effect postulated to derive from oxidative stress. We report here the first
investigation of MPP+-induced oxidative stress in the murine neuroblastoma cell line N2A. Significant cell death was observed following exposure to 0.25 mM MPP+. Markers of oxidative stress included decreased intracellular levels of GSH after 48 h of exposure (85% depletion) as well as an increase in GSSG. Expression of both superoxide dismutase 1 (sod1) and catalase (cat) mRNA was increased, as well the activity of catalase. These cellular effects were, at least partially, reversed by treatment with the natural polyphenol mangiferin. Administration of mangiferin protected N2A cells against MPP+-induced cytotoxicity, restored the GSH content (to 60% of control levels), and down-regulated both sod1 and cat mRNA expression. Together, these results suggest that the protective effect of mangiferin in N2A cells is mediated by the quenching of reactive oxygen intermediates. Therefore, mangiferin could be a useful compound in therapies for degenerative diseases, including Parkinson's disease, in which oxidative stress plays a crucial role.
65
Study On Mango Leaf and Mangiferin Mangiferin protects human peripheral blood lymphocytes against -radiation-induced DNA strand breaks: a fluorescence analysis of DNA unwinding assay
G. Jagetia, V. Venkatesha Department of Radiobiology, Kasturba Medical College, Manipal, India
Ionizing radiations induce free radicals that lead to a cascade of events causing damage to cellular DNA. It is generally accepted that cell death induced by ionizing irradiation is due to DNA double-strand breaks. Therefore, agents that can neutralize free radicals may be able to reduce DNA damage effectively and protect cell death. Effect of 0, 5, 10, 20, 50, or 100 g/mL mangiferin, a glucosylxanthone present in mango (Mangifera indica), has been studied in human peripheral blood lymphocytes (HPBLs) exposed to 3 Gy -radiation by fluorescence analysis of DNA unwinding assay. This assay detects DNA single- and double-strand breaks and alkali-labile sites by their effect on the rate of DNA denaturation in alkali, which was monitored by the fluorescence intensity of an intercalating dye, Hoechst 33258 (bisbenzimide). Estimation of DNA damage by fluorescence analysis of DNA unwinding assay showed that mangiferin as such did not have adverse effect on DNA damage, and it reduced the radiation-induced DNA damage in a concentration-dependent manner. In addition, a maximum undamaged double-stranded DNA was observed for 50 g/mL of mangiferin. Therefore, further experiments were carried out using this concentration, wherein lymphocytes were exposed to 0, 1, 2, 3, or 4 Gy -radiation 30 minutes after mangiferin treatment. Irradiation of HPBLs caused a radiation dose-dependent increase in the DNA strand breaks and a reduction in the undamaged double-stranded DNA, whereas treatment of lymphocytes with 50 g/mL mangiferin before irradiation significantly reduced DNA strand breaks and subsequently enhanced the undamaged double-stranded DNA at 4 hours posttreatment, indicating repair of radiation-induced DNA strand breaks. Mangiferin treatment restored the undamaged double-stranded DNA to almost normal level after 1 Gy irradiation, whereas it was 50% for 4 Gy at 4 hours postirradiation. Our observations suggest that mangiferin reduces initial DNA damage and enhances DNA repair in the HPBLs exposed to 1 to 4 Gy -radiation and could serve as a protector against the radiation-induced DNA damage during planned and unplanned radiation exposures.
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Study On Mango Leaf and Mangiferin Mangiferin protects the streptozotocin-induced oxidative damage to cardiac and renal tissues in rats
Muruganandan S, Gupta S, Kataria M, Lal J, Gupta PK Division of Pharmacology and Toxicology, Indian Veterinary Research Institute, Bareilly District, UP, India
The role of oxidative stress in streptozotocin (STZ)-induced toxicity and its prevention by a xanthone glucoside, mangiferin was investigated. To induce diabetes mellitus, adult male Wistar rats were injected STZ intravenously at 55 mg/kg body weight. The effect of mangiferin (10 and 20 mg/kg, i.p., 28 days) was investigated in STZ-induced diabetic male rats. Insulin-treated rats (6 U/kg, i.p., 28 days) served as positive control. Diabetic rats given normal saline served as negative control. Normal rats that neither received STZ nor drugs served as normal control. On day 28, the diabetic rats showed significant increase in serum creatine phosphokinase (CPK) and total glycosylated haemoglobin. Kidney revealed tubular degeneration and decreased levels of superoxide dismutase (SOD) and catalase (CAT) with an elevation of malonaldehyde (MDA). Cardiac SOD, CAT and lipid peroxidation were significantly increased. Histopathological findings revealed cardiac hypertrophy with haemorrhages. Analysis of erythrocyte revealed significantly elevated levels of MDA with insignificant decrease in CAT and SOD. Repeated intraperitoneal injections of mangiferin (10 and 20 mg/kg) and insulin (6 U/kg) controlled STZ-induced lipid peroxidation and significantly protected the animals against cardiac as well as renal damage. From the study, it may be concluded that oxidative stress appears to play a major role in STZ-induced cardiac and renal toxicity as is evident from significant inhibition of antioxidant defence mechanism in renal tissue or a compensatory increase in antioxidant defence mechanism in cardiac tissue. Intraperitoneal administration of mangiferin exhibited significant decrease in glycosylated haemoglobin and CPK levels along with the amelioration of oxidative stress that was comparable to insulin treatment.
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Study On Mango Leaf and Mangiferin Mangiferin, a natural occurring glucosyl xanthone, increases susceptibility of rat liver mitochondria to calcium-induced permeability transition
Andreu GL, Delgado R, Velho JA, Curti C, Vercesi AE Departamento de Patologia Clnica, Faculdade de Cincias Mdicas, Universidade Estadual de Campinas, Campinas, SP, Brazil
Mitochondrial permeability transition (MPT) is a Ca2+-dependent, cyclosporine A-sensitive, non-selective inner membrane permeabilization induced by a wide range of agents or conditions, which has often been associated with necrotic or apoptotic cell death. When mitochondria isolated from livers of rats treated with the natural occurring glucosyl xanthone mangiferin (40 mg/kg body weight) were exposed in vitro to Ca2+, they underwent CsA, NEM, and ADP-sensitive high amplitude swelling and associated membrane potential dissipation, release of pre-accumulated Ca2+, oxidation of thiol groups, and depletion of GSH, without changes in the NAD(P)H redox state. The same treatment reduced the phosphorylation rate of mitochondria and the resting respiration by around 4 and 11%, respectively, as well as generation of reactive oxygen species (ROS) by organelle. The in vitro exposure of untreated mitochondria to mangiferin plus Ca2+ also resulted in oxidation of thiol groups, in the same way that the compound inhibited the Ca2+-induced peroxidation of mitochondrial membrane lipids. The spectrum of mangiferin during its oxidation by the H2O2/HRP system showed a characteristic absorption peak at 380 nm, which decreased immediately after reaction was started; two isosbestic points at around 336 and 412 nm, with a blue shift in the position of the maxima absorption of mangiferin were observed, suggesting their conversion into one oxidation product. Glutathione abolished this decrease of absorbance, suggesting that the oxidation product of mangiferin forms adducts with GSH. We propose that Ca2+ increases levels of mitochondria-generated ROS, which reacts with mangiferin producing quinoid derivatives, which in turn react with the most accessible mitochondrial thiol groups, thus triggering MPT. It seems probable that the free radical scavenging activity of mangiferin shifts its anti-oxidant protection to the thiol arylation. An interesting proposition is that accumulation of mangiferin quinoid products would take place in cells exposed to an overproduction of ROS, such as cancer cells, where the occurrence of MPT-mediated apoptosis may be a cellular defence mechanism against excessive ROS formation.
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Study On Mango Leaf and Mangiferin Mangiferin protects human peripheral blood lymphocytes against -radiationinduced DNA strand breaks:a fluorescence analysis of DNA unwinding assay
Ganesh Chandra Jagetia,Venkatasubbaiah A.Venkatesha Department of Radiobiology,Kasturba Medical College,Manipal,India
Ionizing radiations induce free radicals that lead to a cascade of events causing damage to cellular DNA.It is generally accepted that cell death induced by ionizing irradiation is due to DNA double-strand breaks.Therefore,agents that can neutralize free radicals may be able to reduce DNA damageeffectively and protect cell death.Effect of 0,5,10,20,50,or100 g/mL mangiferin,a glucosylxanthone present in mango(Mangifera indica),has been studied in human peripheral blood lymphocytes(HPBLs)exposed to 3 Gy -radiation by fluorescence analysis of DNA unwinding assay.This assay detects DNA single-and double-strand breaks and alkali-labile sites by their effect on the rate of DNA denaturation in alkali,which was monitored by the fluorescence intensity of an intercalating dye,Hoechst
33258(bisbenzimide).Estimation of DNA damage by fluorescence analysis of DNA unwinding assay showed that mangiferin as such did not have adverse effect on DNA damage,and it reduced the radiation-induced DNA damage in a concentration-dependent manner.In addition,a maximum undamaged double-stranded DNA was observed for 50 g /mL of mangiferin.Therefore,further experiments were carried out using this concentration,wherein lymphocytes were exposed to 0,1,2,3,or 4 Gy -radiation 30 minutes after mangiferin treatment. Irradiation of HPBLs caused a radiation dose-dependent increase in the DNA strand breaks and a reduction in the undamaged double-stranded DNA,whereas treatment of lymphocytes with 50 g / mL mangiferin before irradiation significantly reduced DNA strand breaks and subsequently enhanced the undamaged double-stranded DNA at 4 hours posttreatment,indicating repair of radiation-induced DNA strand breaks.Mangiferin treatment restored the undamaged double-stranded DNA to almost normal level after 1 Gy irradiation,whereas it was 50%for 4 Gy at 4 hours postirradiation.Our observations suggest that mangiferin reduces initial DNA damage and enhances DNA repair in the HPBLs exposed to 1 to 4 Gy -radiation and could serve as a protector against the radiation-induced DNA damage during planned and unplanned radiation exposures.
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Study On Mango Leaf and Mangiferin Mechanism of Antioxidant Action of Pueraria Glycoside (PG)-1 (an Isoflavonoid) and Mangiferin (a Xanthonoid)
Sato T, Kawamoto A, Tamura A, Tatsumi Y, Fujii T Department of Biochemistry, Kyoto Pharmaceutical University, Japan
The antioxidant activities of pueraria glycoside (PG)-1 (isoflavonoid) and mangiferin (xanthonoid) were studied and compared with PG-3 and daidzein (isoflavonoids) and with wogonin (flavonoid). PG-1 and mangiferin rapidly scavenged 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, and inhibited lipid peroxidation which was initiated enzymatically by reduced nicotinamide adenine dinucleotide phosphate (NADPH) or non-enzymatically by ascorbic acid or Fenton's reagent (H2O2 + Fe2+) in rat liver
microsomes. Wogonin inhibited the enzymatically induced lipid peroxidation but had no scavenging effect on DPPH radical or on the non-enzymatic peroxidation. PG-3 and daidzein did not show any of these effects. Formation of Fe2+ by NADPH-dependent cytochrome P-450 reductase was inhibited by wogonin, but not by PG-1 or mangiferin. PG-1 and mangiferin had no effect on terminating radical chain reaction during the lipid peroxidation in the enzymatic system of microsomes or in the linoleic acid hydroperoxide-induced peroxidation system. These results suggest that PG-1 and mangiferin have an antioxidant activity, probably due to their ability to scavenge free radicals involved in initiation of lipid peroxidation. In contrast, wogonin may affect NADPH-dependent cytochrome P-450 reductase action, since it inhibited only the enzymatically induced lipid peroxidation.
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Study On Mango Leaf and Mangiferin Mechanism of cell death induced by an antioxidant extract of Cratoxylum cochinchinense (YCT) in Jurkat T cells: the role of reactive oxygen species and calcium
Tang SY, Whiteman M, Jenner A, Peng ZF, Halliwell B Department of Biochemistry, Faculty of Medicine, National University of Singapore, Singapore
YCT is a semipurified extract from Cratoxylum cochinchinense that has antioxidant properties and contains mostly mangiferin. We show here that YCT is selectively toxic to certain cell types and investigate the mechanisms of this toxicity in Jurkat T cells. By flow cytometric analyses, we show that YCT causes intense oxidative stress and a rise in cytosolic Ca2+. This is followed by a rise in mitochondrial Ca2+, release of cytochrome c, collapse of Deltapsi(m), a fall in ATP levels, and eventually cell death. The mechanism(s) of intense oxidative stress may involve a plasma membrane redox system, as cell death is inhibited by potassium ferricyanide. Cell death has some features of apoptosis (propidium iodide staining, externalization of phosphatidylserine, limited caspase-3 and -9 activities), but there was no internucleosomal DNA fragmentation.
Mechanism of protective action of mangiferin on suppression of inflammatory response and lysosomal instability in rat model of myocardial infarction
Prabhu S, Narayan S, Devi CS Department of Biochemistry, University of Madras, Guindy Campus, Chennai, India
Lysosomal instability has been suggested as a major factor in the development of cellular injury during myocardial necrosis through the formation of inflammatory mediators. The present study was designed to investigate the effect of mangiferin on lysosomal hydrolases and TNF- production during isoproterenol (ISPH) induced myocardial necrosis in rats. The rats given ISPH (200 mg/kg body weight twice, subcutaneous) for 2 days showed a significant increase in plasma TNF- production, serum and heart lysosomal hydrolases activity. ISPH administration to rats resulted in decreased stability of the membranes, which was reflected by the lowered activity of cathepsin-D and beta-glucuronidase in mitochondrial, nuclear, lysosomal and microsomal fractions. Pretreatment with mangiferin (100 mg/kg body weight, intraperitoneally) for 28 days, significantly prevented the alterations and restored the enzyme activities to near-normal status. These findings demonstrate that mangiferin could preserve lysosomal integrity through decrease in the inflammatory process and hence establish the cardioprotective effect of mangiferin.
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Study On Mango Leaf and Mangiferin Mechanisms of blood glucose-lowering effect of aqueous extract from stems of Kothala himbutu (Salacia reticulata) in the mouse
Im R, Mano H, Matsuura T, Nakatani S, Shimizu J, Wada M Department of Clinical Dietetics and Human Nutrition, Faculty of Pharmaceutical Sciences, Josai University, Saitama, Japan
Ethnopharmacological relevance: Kothala himbutu (Salacia reticulata) is a medicinal plant that has been used in Ayurvedic system of Indian and Sri Lankan traditional medicine to treat diabetes. This study aimed to clarify the mechanism(s) by which aqueous extracts of Kothala himbutu (KTE) stems decreases fasting blood glucose levels. Gene expression profiles were assessed by DNA microarray and RT-PCR analyses of RNA from the liver of KK-Ay diabetic mice administered KTE or control distilled water for 4 weeks, and from cultured liver cells treated with freeze-dried KTE (KTED) or selected phenolic compounds. DNA microarray and RT-PCR analyses revealed that gluconeogenic
fructose-1,6-bisphosphatase (FBP) was decreased compared with the control in KTE-treated KK-Ay mice. RT-PCR analysis using cultured liver cells treated with KTED and/or actinomycin D or cycloheximide, revealed that KTED directly decreased FBP mRNA levels via destabilization of the mRNA. One compound in KTE, mangiferin, was demonstrated to dose-dependently down-regulate FBP mRNA. These findings suggest that the mangiferin in KTE acts directly on liver cells and down-regulates the gluconeogenic pathway through regulation of FBP expression, thereby decreasing fasting blood glucose levels in mice. Our results demonstrate that gluconeogenic gene regulation is one possible mechanism by which KT exerts its effects in traditional diabetic medicine.
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Study On Mango Leaf and Mangiferin Modulation of P450 enzymes by Cuban natural products rich in polyphenolic compounds in rat hepatocytes
Rodeiro I, Donato MT, Lahoz A, Gonzlez-Lavaut JA, Laguna A, Castell JV, Delgado R, Gmez-Lechn MJ Departmento de Investigaciones Biomedicas, Centro de Qumica Farmacutica, Ciudad de la Habana, Cuba
This paper reports cytotoxic effects and changes in the P450 system after exposing rat hepatocytes to four polyphenol-rich products widely used in Cuban traditional medicine (Mangifera indica L. (MSBE), Thalassia testudinum (Tt), Erythroxylum minutifolium and confusum extracts). Effects of mangiferin, the main polyphenol in MSBE, were also evaluated. Cytotoxicity was assayed by the MTT test after exposure of cells to the products (50-1000 g/mL) for 24 or 72 h. The results showed that 500 g/mL MSBE was moderately cytotoxic after 72 h, while mangiferin was not. Marked reductions in cell viability were produced by Erythroxylum extracts at concentrations200 g/mL, whereas only moderate effects were induced by 1000 g/mL Tt. Seven specific P450 activities were evaluated after 48 h exposure of cells to the products. MSBE reduced phenacetin O-deethylation (POD; CYP1A2) activity in a concentration-dependent manner (IC50=190 g/mL). No decreases were observed in other activities. In contrast, mangiferin produced reductions in five P450 activities: IC50 values of 132, 194, >200, 151 and 137 g/ml for POD (CYP1A2), midazolam 1'-hydroxylation (M1OH; CYP3A1), diclofenac 4'-hydroxylation (D4OH; CYP2C6), S-mephenytoin 4'-hydroxylation (SM4OH), and chlorzoxazone 6-hydroxyaltion (C6OH; CYP2E1), respectively. E. minutifolium, E. confusum and Tt extracts produced small reductions in SM4OH and C6OH activities, but no significant changes were noted in the other P450 activities. On the other hand, all the products increased the benzyloxyresorufin O-debenzylation (BROD; CYP2B1) activity, with MSBE, mangiferin or E. minutifolium showing the highest effects (about 2-fold over control). Our results showed in vitro effects of these natural products on P450 systems, possibly leading to potential metabolic-based interactions.
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Study On Mango Leaf and Mangiferin Modulation of rat macrophage function by the Mangifera indica L. extracts Vimang and mangiferin
Garca D, Delgado R, Ubeira FM, Leiro J Departamento de Farmacia, Universidad Central de Las Villas, Villa Clara, Cuba
Vimang is an aqueous extract of Mangiferia indica L., traditionally used in Cuba as an anti-inflammatory, analgesic and antioxidant. In the present study, we investigated the effects of Vimang and of mangiferin (a C-glucosylxanthone present in the extract) on rat macrophage functions including phagocytic activity and the respiratory burst. Both Vimang and mangiferin showed inhibitory effects on macrophage activity: (a) intraperitoneal doses of only 50-250 mg/kg markedly reduced the number of macrophages in peritoneal exudate following intraperitoneal injection of thioglycollate 5 days previously (though there was no significant effect on the proportion of macrophages in the peritoneal-exudate cell population); (b) in vitro concentrations of 0.1-100 g/ml reduced the phagocytosis of yeasts cells by resident peritoneal and thioglycollate-elicited macrophages; (c) in vitro concentrations of 1-50 g/ml reduced nitric oxide (NO) production by thioglycollate-elicited macrophages stimulated in vitro with lipopolysaccharide (LPS) and IFN; and (d) in vitro concentrations of 1-50 g/ml reduced the extracellular production of reactive oxygen species (ROS) by resident and thioglycollate-elicited macrophages stimulated in vitro with phorbol myristate acetate (PMA). These results suggest that components of Vimang, including the polyphenol mangiferin, have depressor effects on the phagocytic and ROS production activities of rat macrophages and, thus, that they may be of value in the treatment of diseases of immunopathological origin characterized by the hyperactivation of phagocytic cells such as certain autoimmune disorder.
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Molecular mechanisms of neuroprotection by two natural antioxidant polyphenols

Campos-Esparza MR, Snchez-Gmez MV, Matute C Departamento de Neurociencias, Facultad de Medicina y Odontologa. Universidad del Pas Vasco, and CIBERNED, Leioa, Spain
Excessive activation of glutamate receptors, or excitotoxicity, contributes to acute and chronic neurological disorders including stroke. We previously showed that two natural polyphenol antioxidants, mangiferin and morin, are neuroprotective in a model of ischemic brain damage. In this study, we analyzed the molecular mechanisms underlying neuroprotection by mangiferin and morin in an in vitro model of excitotoxic neuronal death involving NMDA receptor overactivation. We observed that both polyphenols reduce the formation of reactive oxygen species, activate the enzymatic antioxidant system, and restore the mitochondrial membrane potential. Moreover, both antioxidants inhibit glutamate-induced activation of calpains, normalize the levels of phosphorylated Akt kinase and Erk1/2, as well as of cytosolic Bax, inhibit AIF release from mitochondria, and regulate the nuclear translocation of NF-kappaB. Each of these effects contributes to the substantial reduction of apoptotic neuronal death induced by glutamate. These results demonstrate that mangiferin and morin exhibit excellent antioxidant and antiapoptotic properties, supporting their clinical application as trial neuroprotectors in pathologies involving excitotoxic neuronal death.
New antidiabetic compounds, mangiferin and its glucoside

Ichiki H, Miura T, Kubo M, Ishihara E, Komatsu Y, Tanigawa K, Okada M Tsumura Central Research Laboratories, Tsumura and Co., Ibaraki, Japan
Mangiferin (MF) and its glucosides (mangiferin-7-O-beta-glucoside) (MG) isolated from Anemarrhena asphodeloides Bunge rhizome, were tested for their antidiabetic activity in KK-Ay mice, an animal model of non-insulin-dependent diabetes mellitus (NIDDM). MF and MG lowered the blood glucose level of KK-Ay mice after oral administration. However, no affect on the blood glucose level in normal mice was seen, indicating that MF and MG are useful in treating NIDDM. In addition, MF or MG improved hyperinsulinemia in KK-Ay mice. From these findings, it seems likely that MF and MG exert their its antidiabetic activity by increasing insulin sensitivity.
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Study On Mango Leaf and Mangiferin New antioxidant C-glucosylxanthones from the stems of Arrabidaea samydoides
Pauletti PM, Castro-Gamboa I, Siqueira Silva DH, Young MC, Tomazela DM, Eberlin MN, da Silva Bolzani V NuBBE- Ncleo de Biossntese, Bioensaios e Ecofisiologia de Produtos Naturais, Instituto de Qumica, Universidade Estadual Paulista, Araraquara, SP, Brazil
Three new C-glucosylxanthones, 2-(2'-O-trans-caffeoyl)-C-beta-d-glucopyranosyl-1,3,6,7-tetrahydroxyxanthone (1), 2-(2'-O-trans-cinnamoyl)-C-beta-d-glucopyranosyl-1,3,6,7-tetrahydroxyxanthone (2), and 2-(2'-O-trans-coumaroyl)-C-beta-d-glucopyranosyl-1,3,6,7-tetrahydroxyxanthone (3), were isolated from the stems of Arrabidaea samydoides, in addition to three known C-glucosylxanthones, mangiferin (4), 2-(2'-O-benzoyl)-C-beta-d-glucopyranosyl-1,3,6,7-tetrahydroxyxanthone (5), and muraxanthone (6). Their chemical structures were assigned on the basis of MS and 1D and 2D NMR experiments. Xanthones 1-6 showed moderate freeradical scavenging activity against 1,1-diphenyl-2-picrylhydrazyl DPPH) as well as antioxidant activity evidenced by redox properties measured on ElCD-HPLC.
Neuroprotection by two experimental ischemia
polyphenols
following
excitotoxicity
and
Gottlieb M, Leal-Campanario R, Campos-Esparza MR, Snchez-Gmez MV, Alberdi E, Arranz A, Delgado-Garca JM, Gruart A, Matute C Departamento de Neurociencias, Universidad del Pas Vasco, Leioa, Vizcaya, Spain
Brain ischemia induces neuronal loss which is caused in part by excitotoxicity and free radical formation. Here, we report that mangiferin and morin, two antioxidant polyphenols, are neuroprotective in both in vitro and in vivo models of ischemia. Cell death caused by glutamate in neuronal cultures was decreased in the presence of subMolar concentrations of mangiferin or morin which in turn attenuated receptor-mediated calcium influx, oxidative stress as well as apoptosis. In addition, both antioxidants diminished the generation of free radicals and neuronal loss in the hippocampal CA1 region due to transient forebrain ischemia in rats when administered after the insult. Importantly, neuroprotection by these antioxidants was functionally relevant since treated-ischemic rats performed significantly better in three hippocampal-dependent behavioral tests. Together, these results indicate that mangiferin and morin have potent neuroprotectant activity which may be of therapeutic value for the treatment of acute neuronal damage and disability.
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Study On Mango Leaf and Mangiferin Novel screening assay for antioxidant protection against peroxyl radical-induced loss of protein function
Bertolini F, Novaroli L, Carrupt PA, Reist M LCT-Pharmacochemistry, School of Pharmaceutical Sciences, University of Geneva,University of Lausanne, Switzerland
Oxidative damage to proteins, implicated amongst other in the etiology and progression of Parkinson's disease (PD) and Alzheimer's disease (AD), results in the loss of specific biological protein function. A
simple, sensitive, and cost-effective fluorimetric test to assess the antioxidant capacity of new chemical entities to protect proteins from loss of activity caused by reactive oxygen species (ROS) was developed using alkaline phosphatase (ALP) as model protein. Protein oxidation was induced by
2,2'-azobis(2-methylpropionamidine) dihydrochloride (AAPH) and the decrease in catalytic activity of ALP to hydrolyze 4-methylumbelliferyl phosphate (4-MUP) to fluorescent 4-methylumbelliferone (4-MU) was monitored as a marker of protein degradation. According to their capacity to protect ALP from peroxyl radical-induced activity loss, ten reference antioxidants were divided into three classes, namely efficient (pIC50 > 5 for quercetin, chlorogenic acid, caffeic acid, mangiferin, and resveratrol), intermediate (4 < pIC50 5 for melatonin, trolox, and ascorbic acid), and poor antioxidants (pIC50< 4 for glutathione and D-mannitol). Multifunctional drugs, having the ability to interact with several disease-related targets are of interest in PD. Therefore, the capacity of three catechol-O-methyltransferase (COMT) inhibitors, entacapone, nitecapone, and tolcapone to protect ALP from oxidative damage was also investigated and found to be very similar to the most potent reference antioxidants.
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Study On Mango Leaf and Mangiferin Pharmacokinetic study of free mangiferin in rats by microdialysis coupled with microbore high-performance liquid chromatography and tandem mass spectrometry
Lai L, Lin LC, Lin JH, Tsai TH National Laboratories of Foods and Drugs, Department of Health, Executive Yuan, Taipei, Taiwan
Mangiferin (2-beta-D-glucopyranosyl-1,3,6,7-tetrahydroxyxanthen-9-one) has been isolated from the herbal root of Anemarrhena asphodeloides Bung showing antioxidative, antiviral, and anticancer effect. An in vivo microdialysis sampling method coupled to microbore high-performance liquid chromatography (HPLC) was employed for continuous monitoring of free mangiferin in rat blood. Microdialysis probes were inserted into the jugular vein/right atrium and brain striatum of Sprague-Dawley rats, and mangiferin at doses of 10, 30 or 100 mg/kg were then administered via the femoral vein. Dialysates were collected every 10 min and injected directly into a microbore HPLC system. Mangiferin was separated by a reversed-phase C18 microbore column (1501 mm) from dialysate within 10 min. The mobile phase consisted of acetonitrile-0.05% phosphoric acid-tetrahydrofuran (10:75:15, v/v/v) with a flow-rate of 0.05 ml/min. The wavelength of the UV detector was set at 257 nm. The limit of quantification for mangiferin was 0.05 g/ml and in vivo recovery of mangiferin at concentrations of 1, 5 and 10 g/ml was in range of 37.7-39.8%. The results indicate that the pharmacokinetics of mangiferin at doses of 10-30 mg/kg reveals a linear relation, while doses of 30-100 mg/kg show a nonlinear pharmacokinetic phenomenon. Mangiferin was undetectable in brain dialysate. The proposed method provides a technique for rapid and sensitive analysis of free mangiferin in rat blood and further application in pharmacokinetic study. Furthermore, the metabolites of mangiferin in the rat bile were confirmed by LC electrospray ionization (ESI) tandem mass spectrometry (MS-MS).
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Study On Mango Leaf and Mangiferin Physiological and biochemical changes with special reference to mangiferin and oxidative enzymes level in malformation resistant and susceptible cultivars of mango (Mangifera indica L.)
V.K. Singh Central Institute for Subtropical Horticulture,Rehmankhera,Lucknow,Uttar Pradesh,India
Changes in biophysical attributes, mangiferin and polyphenol oxidase (PPO), catalase and peroxidase activities in malformation resistant mango cultivar Elaichi were studied at various stages of flower development and compared with susceptible cvs. Amrapali, Beauty Mc-lin and Dashehari. Accumulation of mangiferin was maximum (96.0 and 108.0 mg g-1 FW) in Elaichi prior to flower bud differentiation (September) and at full bloom (February), while these were minimum (59.0 and 74.0 mg g-1 FW) in susceptible cv. Beauty Mc-lin. Mangiferin promoted vegetative growth and exhibited inhibitory role on the occurrence of malformation. It was also found that the resistant cultivar had highest activity of PPO as compared to susceptible ones. There was no significant difference in the enzymes catalase and peroxidase activity at early stage of flower differentiation but at flower bud burst stage the catalase activity was enhanced significantly in cv. Elaichi (25.28 unit min-1 g-1 FW) in comparison to Amrapali (16.20 unit min-1 g-1 FW), Beauty Mc-lin (18.39 unit min-1 g-1 FW) and Dashehari (17.50 unit min-1 g-1 FW). The resistant cultivar had high leaf temperature (30.30 degrees C) and diffusion resistance (476.14 m mol m-2 s-1) during the flowering but the rate of transpiration and relative humidity (RH) were high in susceptible cultivars. Results of the present study clearly indicate that level of mangiferin could be considered as a potential biochemical indicator for screening mango genotypes to malformation.
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Study On Mango Leaf and Mangiferin Polyphenols with antiulcerogenic action from aqueous decoction of mango leaves (Mangifera indica L.)
Severi JA, Lima ZP, Kushima H, Brito AR, Santos LC, Vilegas W, Hiruma-Lima CA Pharmacos and Drugs Department, Pharmaceutical Sciences Faculty, So Paulo State University-UNESP, UNESP, Araraquara, SP, Brazil
This study was designed to determine the gastroprotective effect of a Mangifera indica leaf decoction (AD), on different experimental models in rodents. The administration of AD up to a dose of 5 g/kg (p.o.) did not produce any signs or symptoms of toxicity in the treated animals, while significantly decreasing the severity of gastric damage induced by several gastroprotective models. Oral pre-treatment with AD (250, 500 or 1000 mg/kg) in mice and rats with gastric lesions induced by HCl/ethanol, absolute ethanol, non-steroidal anti-inflammatory drug (NSAID) or stress-induced gastric lesions resulted in a significant decrease of said lesions. Phytochemical analyses of AD composition demonstrated the presence of bioactive phenolic compounds that represent 57.3% of total phenolic content in this extract. Two main phenolic compounds were isolated, specifically mangiferin (C-glucopyranoside of 1,3,6,7-tetrahydroxyxanthone) and C-glucosyl-benzophenone (3-C-beta-D-glucopyranosyl-4',2,4,6-tetrahydroxybenzophenone). These findings indicate the potential gastroprotective properties of aqueous decoction from M. indica leaves.
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Study On Mango Leaf and Mangiferin Potential hepatoprotective effects of new Cuban natural products in rat hepatocytes culture
Rodeiro I, Donato MT, Martnez I, Hernndez I, Garrido G, Gonzlez-Lavaut JA, Menndez R, Laguna A, Castell JV, Gmez-Lechn MJ Laboratorio de Farmacologa, Departamento de Investigaciones Biomdicas, Centro de Qumica Farmacutica, 200 y 21, Atabey, Playa, Ciudad de la Habana, Cuba
The protective effects of five Cuban natural products (Mangifera indica L. (MSBE), Erythroxylum minutifolium, Erythroxylum confusum, Thalassia testudinum and Dictyota pinnatifida extracts and mangiferin) on the oxidative damage induced by model toxicants in rat hepatocyte cultures were studied. Cells were pre-incubated with the natural products (5-200 g/mL) for 24 h. Then hepatotoxins (tert-butyl hydroperoxide, ethanol, carbon tetrachloride and lipopolysaccharide) were individually added and post-incubated for another 24 h. After treatments, cell viability was determined using the MTT assay. Mangiferin and MSBE exhibited the highest cytoprotective potential (EC50 between 50 and 125 mg/mL), followed by T. testudinum and Erythroxylum extracts, whereas no significant protective effects was produced by Dictyota extract treatment. Antioxidant properties of the natural products against lipid peroxidation and GSH depletion induced by tert-butyl hydroperoxide were then investigated. The results show that at 36 h pre-treatment of cells with mangiferin or MSBE, concentrations of T. testudinum and Erythroxylum extracts ranging from 25 to 100 mg/mL significantly inhibited lipid peroxidation induced by tert-butyl hydroperoxide (100 and 250 mM) and increased the GSH levels reduced by the toxicant. D. pinnatifida inhibited lipid peroxidation, but did not preserve GSH levels. In conclusion, MSBE, E. minutifolium, E. confusum and T. testudinum extracts and mangiferin showed hepatoprotective activity against induced damage in all the experimental series, where mangiferin and the extracts of MSBE and T. testudinum were the best candidates to inhibit "in vitro" damage to rat hepatocytes. This hepatoprotective effect found could be associated with the antioxidant properties observed for the products.
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Study On Mango Leaf and Mangiferin Protection against septic shock and suppression of tumor necrosis factor and nitric oxide production on macrophages and glia by a standard aqueous extract of Mangifera indica L. (VIMANG). Role of mangiferin isolated from the extract
Garrido G, Delgado R, Lemus Y, Rodrguez J, Garca D, Nez-Sells AJ Laboratorio de Farmacologa, Centro de Qumica Farmacutica, Habana, Cuba
The present study illustrates the effects of a standard aqueous extract, used in Cuba under the brand name of VIMANG, from the stem bark of Mangifera indica L. on the production of tumor necrosis factor (TNF) and nitric oxide (NO) in in vivo and in vitro experiments. In vivo was determined by the action of the extract and its purified glucosylxanthone (mangiferin) on TNF in a murine model of endotoxic shock using Balb/c mice pre-treated with lipopolysaccharide (LPS) 0.125 mg kg-1, i.p. In vitro, M. indica extract and mangiferin were tested on TNF and NO production in activated macrophages (RAW264.7 cell line) and glia (N9 cell line) stimulated with LPS (10ng ml-1) and interferon (IFN, 2U ml-1). M. indica extract reduced dose-dependently TNF production in the serum (ED50 = 64.5 mg kg-1) and the TNF mRNA expression in the lungs and livers of mice. Mangiferin also inhibited systemic TNF at 20 mg kg-1. In RAW264.7, the extract inhibited TNF (IC50 = 94.1 g ml-1) and NO (IC50 = 64.4 g ml-1). In glia the inhibitions of the extract were IC50 = 76.0 g ml-1 (TNF) and 84.0 g ml-1 (NO). These findings suggest that the anti-inflammatory response observed during treatment with M. indica extract must be related with inhibition of TNF and NO production. Mangiferin, a main component in the extract, is involved in these effects. The TNF and NO inhibitions by M. indica extract and mangiferin on endotoxic shock and glia are reported here for the first time.
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Study On Mango Leaf and Mangiferin Protective effect of Mangifera indica L. polyphenols on human T lymphocytes against activation-induced cell death
Hernandez P, Rodriguez PC, Delgado R, Walczak H Department of Biomedical Research, Center of Pharmaceutical Chemistry, Havana, Cuba
Activation-induced cell death (AICD) plays an important role in maintenance of peripheral lymphocyte homeostasis. Reactive oxygen species (ROS) combined with simultaneous calcium (Ca2+) influx into the cytosol are required for induction of AICD. The extract obtained from the stem bark of Mangifera indica L. has shown to protect T cells from in vitro AICD. This extract is rich in polyphenolic compounds, the three main components of which are mangiferin (MA), catechin (C) and epicatechin (EC). The present study has focused on the possible contribution of the polyphenols MA, C and EC to the demonstrated protective effect of M. indica extract on in vitro human T cell AICD. Our results show that these polyphenols diminished the increase of intracellular ROS and free Ca2+ induced by T cell receptor (TCR) triggering. In addition, these polyphenols attenuated AICD. Our findings suggest that the T cell survival effect of M. indica extract is mediated, at least in part, by its main polyphenols.
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Study On Mango Leaf and Mangiferin Protective effects of a standard extract of Mangifera indica L. (VIMANG) against mouse ear edemas and its inhibition of eicosanoid production in J774 murine macrophages
Garrido G, Gonzlez D, Lemus Y, Delporte C, Delgado R Laboratorio de Farmacologa, Centro de Qumica Farmacutica, Habana, Cuba
A standard aqueous extract of Mangifera indica L., used in Cuba as antioxidant under the brand name VIMANG, was tested in vivo for its anti-inflammatory activity, using commonly accepted assays. The standard extract of M. indica, administered orally (50-200mg/kg body wt.), reduced ear edema induced by arachidonic acid (AA) and phorbol myristate acetate (PMA) in mice. In the PMA model, M. indica extract also reduced myeloperoxidase (MPO) activity. In vitro studies were performed using macrophage cell line J774 stimulated with pro-inflammatory stimuli lipopolysaccharide-interferon gamma (LPS-IFN) or calcium ionophore A23187 to determine prostaglandin PGE2 or leukotriene LTB4 release, respectively. The extract inhibited the induction of PGE2 and LTB4 with IC50 values of 21.7 and 26.0g/ml, respectively. Mangiferin (a glucosylxanthone isolated from the extract) also inhibited these AA metabolites (PGE2, IC50 value=17.2g/ml and LTB4, IC50 value=2.1g/ml). These results represent an important contribution to the elucidation of the mechanism involved in the anti-inflammatory and anti-nociceptive effects reported for the standard extract of M. indica VIMANG.
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Study On Mango Leaf and Mangiferin Protective effects of Mangifera indica L. extract (Vimang), and its major component mangiferin, on iron-induced oxidative damage to rat serum and liver
Pardo-Andreu GL, Barrios MF, Curti C, Hernndez I, Merino N, Lemus Y, Martnez I,Riao A, Delgado R. Departamento de Investigaciones Biomdicas, Centro de Qumica Farmacutica, Calle 200, Esq. 21, Playa, Ciudad de La Habana, Cuba
In vivo preventive effects of a Mangifera indica L extract (Vimang) or its major component mangiferin on iron overload injury have been studied in rats given respectively, 50, 100, 250 mg kg-1 body weight of Vimang, or 40 mg kg1 body weight of mangiferin, for 7 days prior to, and for 7 days following the administration of toxic amounts of iron-dextran. Both Vimang or mangiferin treatment prevented iron overload in serum as well as liver oxidative stress, decreased serum and liver lipid peroxidation, serum GPx activity, and increased serum and liver GSH, serum SOD and the animals overall antioxidant condition. Serum iron concentration was decreased although at higher doses, Vimang tended to increase it; percent tranferrin saturation, liver weight/body mass ratios, liver iron content was decreased. Treatment increased serum iron-binding capacity and decreased serum levels of aspartate-amine transferase (ASAT) and alanine-amine transferase (ALAT), as well as the number of abnormal Kupffer cells in iron-loaded livers. It is suggested that besides acting as antioxidants, Vimang extract or its mangiferin component decrease liver iron by increasing its excretion. Complementing earlier in vitro results from our group, it appears possible to support the hypothesis that Vimang and mangiferin present therapeutically useful effects in iron overload related diseases.
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Study On Mango Leaf and Mangiferin Protective effects of Mangifera indica L. extract, mangiferin and selected antioxidants against TPA-induced biomolecules oxidation and peritoneal macrophage activation in mice
Snchez GM, Re L, Giuliani A, Nez-Sells AJ, Davison GP, Len-Fernndez OS Centre for Research and Biological Evaluation, Pharmacy Institute, Havana University, Havana, Cuba
We compared the protective abilities of Mangifera indica L. stem bark extract (Vimang) 50-250 mgkg-1, mangiferin 50 mgkg-1, vitamin C 100 mgkg-1, vitamin E 100 mgkg-1and beta -carotene 50 mgkg-1 against the 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced oxidative damage in serum, liver, brain as well as in the hyper-production of reactive oxygen species (ROS) by peritoneal macrophages. The treatment of mice with Vimang, vitamin E and mangiferin reduced the TPA-induced production of ROS by the peritoneal macrophages by 70, 17 and 44%, respectively. Similarly, the H2O2 levels were reduced by 55-73, 37 and 40%, respectively, when compared to the control group. The TPA-induced sulfhydryl group loss in liver homogenates was attenuated by all the tested antioxidants. Vimang, mangiferin, vitamin C plus E and beta -carotene decreased TPA-induced DNA fragmentation by 46-52, 35, 42 and 17%, respectively, in hepatic tissues, and by 29-34, 22, 41 and 17%, in brain tissues. Similar results were observed in respect to lipid peroxidation in serum, in hepatic mitochondria and microsomes, and in brain homogenate supernatants. Vimang exhibited a dose-dependent inhibition of TPA-induced biomolecule oxidation and of H2O2 production by peritoneal macrophages. Even if Vimang, as well as other antioxidants, provided significant protection against TPA-induced oxidative damage, the former lead to better protection when compared with the other antioxidants at the used doses. Furthermore, the results indicated that Vimang is bioavailable for some vital target organs, including liver and brain tissues, peritoneal exudate cells and serum. Therefore, we conclude that Vimang could be useful to prevent the production of ROS and the oxidative tissue damages in vivo.
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Study On Mango Leaf and Mangiferin Protective role of mangiferin against Benzo(a)pyrene induced lung carcinogenesis in experimental animals
Rajendran P, Ekambaram G, Sakthisekaran D Department of Medical Biochemistry, Dr. ALM PG Institute of Basic Medical Sciences, University of Madras, Taramani, Chennai, India
In recent years, considerable emphasis has been focused on identifying new chemopreventive agents which could be useful for the human population. In the present study, we examined the protective role of mangiferin during experimental lung carcinogenesis with reference to its effect on DNA-damage and the detoxification enzyme system. The activities of detoxifying enzymes such as glutathione transferase (GST), quinone reductase (QR) and uridin 5'-diphosphate-glucuronosyl transferase (UDP-GT) were found to be decreased while the lipid peroxidation level was increased in the lung cancer bearing animals. Supplementation of mangiferin (100 mg/kg b.wt) enhanced the detoxification enzymes and reduced DNA damage as determined by single cell electrophoresis. Furthermore, the DNA-protein cross links which was found to be high in lung cancer bearing animals was also modulated upon supplementation with mangiferin. Our present results explain the unique association between the anti-oxidant effect of mangiferin and ultimately the capability of mangiferin to prevent cancer.
Studies on palauan medicinal herbs. II. Activation of mouse macrophages RAW 264.7 by Ongael, leaves of Phaleria cumingii (Meisn.) F. Vill. and its acylglucosylsterols
Matsuda H, Tokunaga M, Iwahashi H, Naruto S, Yagi H, Masuko T, Kubo M School of Pharmaceutical Sciences, Kinki University, Osaka, Japan
The extract of Ongael [leaves of Phaleria cumingii (MEISN.) F. VILL.], a Palauan medicinal herb, enhanced an in vitro phagocytic activity of mouse macrophages RAW 264.7 cells (RAW 264.7). Activity-guided fractionation of the Ongael extract by the in vitro phagocytosis assay using RAW 264.7 led to the isolation of a mixture of acylglucosylsterols (1) as an active constituent along with other inactive constituents, tetracosanol and mangiferin. On the basis of chemical modifications and spectral analyses, the compound 1 was deduced to be a mixture of the known 3-O-(6-O-acyl-beta-D-glucosyl)-beta-sitosterols, the acyl moiety being mainly palmitoyl (57%), oleoyl (12%) and -linolenoyl (12%) with small amount of stearoyl (7%) and linoleoyl (4%).
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Radioprotection by mangiferin in DBAxC57BL mice: a preliminary study

Jagetia GC, Baliga MS Department of Radiobiology, Kasturba Medical College Manipal, Karnataka, India
The radioprotective effects of various concentrations (0, 0.25, 0.5, 1, 2, 5, 10, 17.5, 25, 50, 75 and 100 mg/kg b.wt.) of mangiferin (MGN) was studied in the DBAxC57BL mice whole body exposed to 10 Gy of -irradiation. Treatment of mice with different doses of MGN, one hour before irradiation reduced the symptoms of radiation sickness and delayed the onset of mortality when compared with the non-drug treated irradiated controls. The radioprotective action of MGN increased in a dose dependent manner up to 2mg/kg and declined thereafter. The highest radioprotective effect was observed at 2mg/kg MGN, where greatest number of animals survived against the radiation-induced mortality. The administration of 0.5, 1, 2, 5, 10 and 17.5 mg/kg MGN reduced the radiation-induced gastrointestinal death as evident by a greater number of survivors up to 10 days in this group when compared with the DDW + 10 Gy irradiation group. A similar effect of MGN was observed for the radiation-induced bone marrow deaths also. Our study demonstrates that mangiferin, a gluosylxanthone, present in the Mangifera indica protected mice against the radiation-induced sickness and mortality and the optimum protective dose of 2mg/kg was 1/200 of LD50 dose (400 mg/kg) of MGN. The administration of 400 mg/kg MGN induced 50% mortality, therefore LD50 of the drug was considered to be 400 mg/kg.
The suppressive effect of mangiferin with exercise on blood lipids in type 2 diabetes
Miura T, Iwamoto N, Kato M, Ichiki H, Kubo M, Komatsu Y, Ishida T, Okada M,Tanigawa K Department of Clinical Nutrition, Suzuka University of Medical Science, Mie, Japan
The effect of mangiferin (MF) with exercise on bood lipids was studied in KK-Ay mice, an animal model of type 2 diabetes. MF (30 mg/kg) reduced the blood cholesterol (p<0.05) and triglyceride level (p<0.01) of KK-Ay mice with exercise 2 weeks after oral administration when compared with the control group. Diabetes also often has elevated lipid levels. Therefore, it may be that MF has beneficial effects on hyperlipidemia in type 2 diabetes.
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Study On Mango Leaf and Mangiferin Release of intermediate reactive hydrogen peroxide by macrophage cells activated by natural products
Moreira RR, Carlos IZ, Vilega W Departamento de Principios Ativos Naturais e Toxicologia Brasil, Facuildade de Ciencias Farmaceuticas, Universidade Estadual Paulista Jlio de Mesquita Filho,Araraquara, So Paulo.Brazil
By determining the hydrogen peroxide (H2O2) released in cultures of peritoneal macrophage cells from Swiss mice, we evaluated the action of 27 vegetable compounds (pristimerin, tingenone, jatrophone, palustric acid, lupeol, cladrastin, ocoteine, boldine, tomatine, yohimbine, reserpine, escopoletin, esculine, plumericin, diosgenin, deoxyschizandrin, p-arbutin, mangiferin, and others) using a 2 mg/ml solution of each compound (100 g/well). Macrophages are cells responsible for the development of the immunological response reaction, liberating more than one hundred compounds into the extracellular environment. Among these are the various cytokines and the intermediate compounds of nitrogen (NO) and oxygen (H2O2). This coordinated sequence of biochemical reactions is known as the "oxidative burst." When we compared the results with those obtained with zymosan (an important stimulator of H2O2) we observed that the compounds showing the highest activity were substances 2 (tingenone), 16 (reserpine) and 20. Other substances such as compounds 1, 4, 5, 6, 8, 12, 13, 14, 15, 17, 19, 23, 24, 26, and 27 also showed a certain activity, but with less intensity than the aforementioned ones. Compounds 3, 7, 9, 10, 11, 18, 21, 22 and 25 presented no activity. These results suggest that natural products (mainly tingenone and reserpine and others) with different chemical structures are strong immunological modulators. However, further tests are needed to determine the 'oxidative burst' in future studies.
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Study On Mango Leaf and Mangiferin Role of mangiferin on biochemical alterations and antioxidant status in isoproterenol-induced myocardial infarction in rats
Prabhu S, Jainu M, Sabitha KE, Devi CS Department of Biochemistry, University of Madras, Guindy Campus, Chennai, India
The current study dealt with the protective role of mangiferin, a polyphenol from Mangifera indica Linn. (Anacardiaceae), on isoproterenol (ISPH)-induced myocardial infarction (MI) in rats through its antioxidative mechanism. Subcutaneous injection of ISPH (200 mg/kg body weight in 1 ml saline) to rats for 2 consecutive days caused myocardial damage in rat heart, which was determined by the increased activity of serum lactate dehydrogenase (LDH) and creatine phosphokinase isoenzymes (CK-MB), increased uric acid level and reduced plasma iron binding capacity. The protective role of mangiferin was analyzed by triphenyl tetrazolium chloride (TTC) test used for macroscopic enzyme mapping assay of the ischemic myocardium. The heart tissue antioxidant enzymes such as superoxide dismutase, catalase, glutathione peroxidase, glutathione transferase and glutathione reductase activities, non-enzymic antioxidants such as cerruloplasmin, Vitamin C, Vitamin E and glutathione levels were altered in MI rats. Upon pretreatment with mangiferin (100 mg/kg body weight suspended in 2 ml of dimethyl sulphoxide) given intraperitoneally for 28 days to MI rats protected the above-mentioned parameters to fall from the normal levels. Activities of heart tissue enzymic antioxidants and serum non-enzymic antioxidants levels rose significantly upon mangiferin administration as compared to ISPH-induced MI rats. From the present study it is concluded that mangiferin exerts a beneficial effect against ISPH-induced MI due to its antioxidant potential, which regulated the tissues defense system against cardiac damage.
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Study On Mango Leaf and Mangiferin Salacia oblonga extract increases glucose transporter 4-mediated glucose uptake in L6 rat myotubes: Role of mangiferin
Girn MD, Sevillano N, Salto R, Haidour A, Manzano M, Jimnez ML, Rueda R, Lpez-Pedrosa JM Department of Biochemistry and Molecular Biology II, School of Pharmacy, University of Granada, Campus de Cartuja sn, Granada, Spain
To evaluate if the antidiabetic properties of Salacia oblonga extract are mediated not only by inhibiting intestinal -glycosidases but also by enhancing glucose transport in muscle and adipose cells. S. oblonga extract effects on 2-deoxy-D-glucose uptake were assayed in muscle L6-myotubes and 3T3-adipocytes. In L6-myotubes, the amount and translocation of glucose transporters were assayed. A fractionation of the extract was carried out to identify the active compounds. Furthermore, we analyzed the phosphorylation status of key components of signaling pathways that are involved in the molecular mechanisms regulating glucose uptake. S. oblonga extract increased 2-deoxy-d-glucose uptake by 50% in L6-myotubes and 3T3-adipocytes. In L6-myotubes, the extract increased up to a 100% the GLUT4 content, activating GLUT4 promoter transcription and its translocation to the plasma membrane. Mangiferin was identified as the bioactive compound. Furthermore, mangiferin effects were concomitant with the phosphorylation of 5'-AMP-activated protein kinase without the activation of PKB/Akt. The effect of mangiferin on 2-deoxy-d-glucose uptake was blocked by GW9662, an irreversible PPAR- antagonist. S. oblonga extract and mangiferin may exert their antidiabetic effect by increasing GLUT4 expression and translocation in muscle cells. These effects are probably mediated through two independent pathways that are related to 5'-AMP-activated protein kinase and PPAR-.
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Salacia oblonga improves cardiac fibrosis and inhibits postprandial hyperglycemia in obese Zucker rats
Li Y, Peng G, Li Q, Wen S, Huang TH, Roufogalis BD, Yamahara J Herbal Medicines Research and Education Center, Faculty of Pharmacy A15, The University of Sydney, NSW, Australia
Diabetes has a markedly greater incidence of cardiovascular disease than the non-diabetic population. The heart shows a slowly developing increase in fibrosis in diabetes. Extended cardiac fibrosis results in increased myocardial stiffness, causing ventricular dysfunction and, ultimately, heart failure. Reversal of fibrosis may improve organ function survival. Postprandial hyperglycemia plays an important role in the development of type 2 diabetes and cardiovascular complications, and has been proposed as an independent risk factor for cardiovascular diseases. Salacia oblonga (S.O.) is traditionally used in the prevention and treatment of diabetes. We investigated the effects of its water extract on cardiac fibrosis and hyperglycemia in a genetic model of type 2 diabetes, the obese Zucker rat (OZR). Chronic administration of the extract markedly improved interstitial and perivascular fibrosis in the hearts of the OZR. It also reduced plasma glucose levels in non-fasted OZR, whereas it had little effect in the fasted animals, suggesting inhibition of postprandial hyperglycemia in type 2 diabetic animals, which might play a role in improvement of the cardiac complications of OZR. Furthermore, S.O. markedly suppressed the overexpression of mRNAs encoding transforming growth factor betas 1 and 3 in the OZR heart, which may be an important part of the overall molecular mechanisms. S.O. dose-dependently inhibited the increase of plasma glucose in sucrose-, but not in glucose-loaded mice. S.O. demonstrated a strong inhibition of -glucosidase activity in vitro, which is suggested to contribute to the improvement of postprandial hyperglycemia.
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Salacia oblonga root decreases cardiac hypertrophy in Zucker diabetic fatty rats: inhibition of cardiac expression of angiotensin II type 1 receptor
Huang TH, He L, Qin Q, Yang Q, Peng G, Harada M, Qi Y, Yamahara J, Roufogalis BD, Li Y Faculty of Pharmacy, University of Sydney, Australia
We investigated the effect of the water extract of Salacia oblonga (SOE), an ayurvedic antidiabetic and antiobesity medicine, on obesity and diabetes-associated cardiac hypertrophy and discuss the role of modulation of cardiac angiotensin II type 1 receptor (AT1) expression in the effect. SOE (100 mg/kg) was given orally to male Zucker diabetic fatty (ZDF) rats for 7 weeks. At the end-point of the treatment, the hearts and left ventricles were weighed, cardiomyocyte cross-sectional areas were measured, and cardiac gene profiles were analysed. On the other hand, angiotensin II-stimulated embryonic rat heart-derived H9c2 cells and neonatal rat cardiac fibroblasts were pretreated with SOE and one of its prominent components mangiferin (MA), respectively. Atrial natriuretic peptide (ANP) mRNA expression and protein synthesis and [3H]thymidine incorporation were determined. SOE-treated ZDF rats showed less cardiac hypertrophy (decrease in weights of the hearts and left ventricles and reduced cardiomyocyte cross-sectional areas). SOE treatment suppressed cardiac overexpression of ANP, brain natriuretic peptide (BNP) and AT1 mRNAs and AT1 protein in ZDF rats. SOE (50-100 g/ml) and MA (25 Mol) suppressed angiotensin II-induced ANP mRNA overexpression and protein synthesis in H9c2 cells. They also inhibited angiotensin II-stimulated [3H]thymidine incorporation by cardiac fibroblasts. Our findings demonstrate that SOE decreases cardiac hypertrophy in ZDF rats, at least in part by inhibiting cardiac AT1 overexpression. These studies provide insights into a potential cardioprotective role of a traditional herb, which supports further clinical evaluation in obesity and diabetes-associated cardiac hypertrophy.
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Salacia reticulata and its polyphenolic constituents with lipase inhibitory and lipolytic activities have mild antiobesity effects in rats
Yoshikawa M, Shimoda H, Nishida N, Takada M, Matsuda H Kyoto Pharmaceutical University, Misasagi, Yamashina-ku, Japan
Salacia (S.) reticulata, a Hippocrateaceae plant distributed in Sri Lankan and Indian forests, has been used as a supplementary food in Japan to prevent obesity and diabetes. We examined the antiobesity effects of the hot water-soluble extract (SRHW) from the roots of S. reticulata using obese rat models and an in vitro study. Body weight (P = 0.07) and periuterine fat storage (P = 0.10) in female Zucker fatty rats (8-9 wk old) tended to be suppressed by oral administration of SRHW (125mg/kg) for 27 d. Male rats fed a high fat diet were not affected by SRHW. Furthermore, SRHW inhibited porcine pancreatic lipase (PL), rat adipose tissue-derived lipoprotein lipase (LPL) and glycerophosphate dehydrogenase (GPDH) activities with 50% inhibitory concentrations (IC50) of 264 (95% confidence limits: 203-393) mg/L, 15 (12-18) mg/L and 54 (35-85) mg/L, respectively, but did not inhibit hormone-sensitive lipase activity in rat adipose tissue. Next, we examined the effects of polyphenols, di- and triterpenes and salacinol isolated from the roots of S. reticulata on lipid metabolizing enzymes and lipolysis. (-)-Epigallocatechin and (-)-epicatechin-(4beta8)(-)-4'-O-methylepigallocatechin inhibited PL activity with IC50 of 88 (not calculated) and 68 (26-122) mg/L, respectively. (-)-Epicatechin, 3beta, 22beta-dihydroxyolean-12-en-29-oic acid and the tannin fraction inhibited LPL activity with IC50 of 81 (54-214), 89 (62-214) and 35 (24-62) mg/L. Only the tannin fraction inhibited GPDH activity with an IC50 of 6.8 (3.4-10.9) mg/L. These constituents may be involved in the lipase and GPDH inhibitory activities of SRHW. On the other hand, SRHW at 100 mg/L tended to enhance lipolysis in rat adipocytes (P = 0.06). Significant lipolytic effects were exerted by mangiferin, (-)-4'-O-methylepigallocatechin and maytenfolic acid at 100 mg/L (P < 0.01). In conclusion, polyphenolic compounds may be involved in the antiobesity effects of SRHW in rats through inhibition of fat metabolizing enzymes (PL, LPL and GPDH) and enhanced lipolysis.
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Study On Mango Leaf and Mangiferin Scavenger effect of a mango (Mangifera indica L.) food supplement's active ingredient on free radicals produced by human polymorphonuclear cells and hypoxanthine-xanthine oxidase chemiluminescence systems
Gabino Garrido,Deyarina Gonzalez,Cheyla Romay,lberto J. Nunez-Selles,Rene Delgado Departamento de Farmacologia,Centro Nacional de Investigaciones Cientificas,La Habana,Cuba
The in vitro antioxidant and free radical scavenging properties of a stem bark aqueous extract of mango tree {Mangifera indica L.), whose formulations are used in Cuba as food supplements under the brand name of Vimang, were studied. Luminol-enhanced chemiluminescence was used to elucidate the effect of this extract on the generation of reactive oxygen species in PMA- or zymosan-stimulated human polymorphonuclear leukocytes and on superoxide radicals generated in the hypoxanthine-xanthine oxidase reaction. Chemiluminescence was reduced in a dose-dependent manner at extract concentrations from 5 to 100 g/ml, most probably by inhibiting the superoxide generation reaction. Part of this M. indica extract antioxidant activity could be ascribed to the presence of mangiferin as its main component.
Spectroscopic investigation of interaction between mangiferin and bovine serum albumin

Lin H, Lan J, Guan M, Sheng F, Zhang H College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou,China
The mechanism of interaction between mangiferin (MA) and bovine serum albumin (BSA) in aqueous solution was investigated by fluorescence spectra, synchronous fluorescence spectra, absorbance spectra and Fourier transform infrared (FT-IR) spectroscopy. The binding constants and binding sites of MA to BSA at different reaction times were calculated. And the distance between MA and BSA was estimated to be 5.20 nm based on Fster's theory. In addition, synchronous fluorescence and FT-IR measurements revealed that the secondary structures of the protein changed after the interaction of MA with BSA. As a conclusion, the interaction between the anti-diabetes Chinese medicine MA and BSA may provide some significant information for the mechanism of the traditional chinese medicine MA on the protein level to cure diabetes or other diseases.
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Study On Mango Leaf and Mangiferin Swertia chirayita mediated modulation of interleukin-1beta, interleukin-6, interleukin-10, interferon-, and tumor necrosis factor- in arthritic mice
Kumar IV, Paul BN, Asthana R, Saxena A, Mehrotra S, Rajan G. Immunobiology Laboratory, Industrial Toxicology Research Centre, Mahatma Gandhi Marg, Lucknow, India
The effect of aqueous extract of Swertia chirayita stem on the pro- and anti-inflammatory cytokines balance in primary joint synovium of adjuvant-induced arthritic mice has been studied. The level of pro-inflammatory cytokines was found elevated in the joint synovium of arthritic mice in comparison to normal joints. Administration of S. chirayita extract in varying doses through the oral route did not modulate the proinflammatory cytokines on day 2. In contrast, by day 12, a dose dependent (0, 11.86 and 23.72 mg/kg body weight) reduction of tumor necrosis factor- (INF-) interleukin-1beta, (IL-) and interferon-, (IFN-) and elevation of Interleukin-10 (IL-10) was observed in the joint homogenates of arthritic mice. Interleukin-6 (IL-6) was not down regulated in joint homogenate of arthritic mice at the dose 11.86 mg/kg but at higher doses (23.72 and 35.58 mg/kg) significant reduction was observed. The aqueous extract was found to possess two polar compounds, amerogentin and mangiferin but was devoid of swerchirin, chiratol, methyl swetianin, and swertanone. Mangiferin has been reported to possess potent anti-inflammatory property and we presume its presence in the aqueous extract of S. chirayita is responsible for reducing TNF-, IL-1, IL-6, and IFN- and/or elevating IL-10 in the joint homogenates of arthritic mice on day 12. This study will help in our understanding of the mechanism of anti-inflammatory action of S. chirayita in the light of proinflammatory and anti-inflammatory cytokine balance.
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Study On Mango Leaf and Mangiferin The inhibitory effects of mangiferin, a naturally glucosylxanthone, in bowel carcinogenesis of male F344 rats occurring
Yoshimi N, Matsunaga K, Katayama M, Yamada Y, Kuno T, Qiao Z, Hara A, Yamahara J, Mori H Department of Pathology, Gifu University School of Medicine, Gifu, Japan
Mangiferin,1,3,6,7-tetrahydroxyxanthone-C2-beta-D-glucoside, is one of xanthone derivatives and C-glucosylxanthones, is widely distributed in higher plants and is one of constituents of folk medicines. Recent studies showed that mangiferin has a potential as an anti-oxidant and an anti-viral agent. In this study, we examined the effects of mangiferin in rat colon carcinogenesis induced by chemical carcinogen, azoxymethane (AOM). We performed two experiments: a short-term assay to investigate the effects of mangiferin on the development of preneoplastic lesions by AOM, aberrant crypt foci (ACF), and the following long-term assay for the influence of mangiferin on tumorigenesis induced by AOM. In the short-term assay, 0.1% mangiferin in a diet significantly inhibited the ACF development in rats treated with AOM compared to rats treated with AOM alone (64.622.0 vs. 108.343.0). In the long-term assay, the group treated with 0.1% mangiferin in initiation phase of the experimental protocol had significantly lower incidence and multiplicity of intestinal neoplasms induced by AOM (47.3 and 41.8% reductions of the group treated with AOM alone for incidence and multiplicity, respectively). In addition, the cell proliferation in colonic mucosa was reduced in rats treated with mangiferin (65-85% reductions of the group treated with AOM alone). These results suggest that mangiferin has potential as a naturally-occurring chemopreventive agent.
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The variation in cytoplasmic distribution of mouse peritoneal macrophage during phagocytosis modulated by mangiferin, an immunomodulator
De A, Chattopadhyay S. Department of Life Science and Biotechnology, Jadavpur University, Kolkata, India
The peritoneal MPhi in the immunological defense removes foreign particles, pathogenic or otherwise, by phagocytosis, and shows movement in search of the target. The macrophage (MPhi) appears in various shapes and sizes, spherical, flattened spindle-shaped, amoeboid, polygonal and with very long extension. On activation, the MPhi shows changes in cell shape, cytokinesis and development of intercellular contacts. A dynamic redistribution of cytoskeleton with cytoplasmic spread and/or extensions occurs with immunomodulators, like Mangiferin (1,3,6,7-tetrahy -droxyxanthone-C2-beta-D-glucoside). The MPhi isolated from the peritoneal fluid of BALB/c mice pretreated with mangiferin and saline control, on challenge, shows redistribution of cytoplasm with variable morphology. Using the image analyses technique for pattern recognition of individual shapes of MPhi leads to the observation of clustering of cells in the coverslip culture. The stimulation in phagocytosis due to mangiferin, shows cytoplasmic spread, long extensions and intercellular contacts. The individual variations in the cytoplasmic redistribution are due to changes in the balance between the cellular surface area and the long extensions where the shape and nature of the phagocytic particle and the type of MPhi are determinants.
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Study On Mango Leaf and Mangiferin Timosaponin AIII, a saponin isolated from Anemarrhena asphodeloides, ameliorates learning and memory deficits in mice
Lee B, Jung K, Kim DH. Department of Life and Nanopharmaceutical Sciences, and Department of Pharmaceutical Science, Kyung Hee University, Seoul, Republic of Korea.
Anemarrhena asphodeloides Bunge (AA, family Liliaceae), which primarily contains xantones, such as mangiferin, and steroidal saponins, such as timosaponin AIII and sarsasapogenin, has been used as an anti-pyretic, anti-inflammatory, anti-diabetic, anti-platelet aggregation, and anti-depressant agent in traditional Chinese medicine. In the present study, the memory-enhancing effects of these saponins were investigated in scopolamine-treated mice. Among saponins, timosaponin AIII (TA3) significantly reversed the scopolamine-induced deficits in a passive avoidance test and in the Morris water maze test. TA3 also increased hippocampal acetylcholine levels in scopolamine-treated mice and dose-dependently inhibited acetylcholinesterase (AChE) activity (IC50 value, 35.4 M). When TA3 (50 mg/kg) was orally administered to mice and its blood concentration was measured by liquid chromatography and tandem mass spectrometry, the Cmax of TA3 occurred 4-6 h after TA3 treatment. The memory-enhancing effect of TA3 was greater when it was administered 5 h before the acquisition trial than 1 h before. Scopolamine treatment in mice increased brain levels of TNF- and IL-1expression. However, treatment with TA3 and scopolamine inhibited the increase of TNF- and IL-1expression. These results suggest that scopolamine may cause learning and memory deficits that are further complicated by inflammation. TA3 also inhibited the activation of NF-kappaB signaling in BV-2 glia and in SK-N-SH neuroblastoma cells induced with TNF- or scopolamine. Nevertheless, TA3 may ameliorate memory deficits, mainly by inhibiting AChE.
99
Two proteins, Mn2+, and low molecular cofactor are required for C-glucosyl-cleavage of mangiferin
Sanugul K, Akao T, Nakamura N, Hattori M Institute of Natural Medicine, Toyama Medical and Pharmaceutical University, Toyama, Japan
C-Glucosides, in which sugars are attached to the aglycone by carbon-carbon bonds, are generally resistant to acid and enzyme hydrolysis. The C-glucosyl bond of mangiferin, a xanthone C-glucoside, was cleaved by anaerobic incubation with a human intestinal bacterium, Bacteroides sp. MANG, to give norathyriol. A cell-free extract obtained by sonication of B. sp. MANG demonstrated cleaving activity for mangiferin to norathyriol by adding NADH, diaphorase, and dithiothreitol. Both high molecular weight (>10 k) and low molecular weight (<10 k) fractions obtained from the cell-free extract were required for the activity. MnCl2 was necessary for the activity, but other metal ions were not. By purification of the high molecular weight fraction using DEAE-cellulose and Phenyl Sepharose column chromatography, two fractions, designated as proteins A and B, were separated and required for the activity. Neither protein A nor protein B alone showed any activity. This is the first report describing a C-glucosyl-cleaving enzyme from human intestinal bacterium that seems to involve a novel enzyme mechanism.
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Study On Mango Leaf and Mangiferin Utilization of mango peels as a source of pectin and polyphenolics
Nicolai Berardini,Matthias Knodler,Andreas Schieber*,Reinhold Carle Hohenheim University, Institute of Food Technology,Section Plant Foodstuff Technology, Stuttgart, Germany
Two different options for the combined recovery of pectin and phenolic compounds from mango peels, a byproduct of industrial mango processing, were developed. After extraction of dried mango peels with diluted sulfuric acid, the phenolic compounds were adsorbed using a styrene-divinylbenzene copolymerisate resin, and pectin was obtained from the effluent by precipitation with ethanol. Phenolic compounds were recovered from the resin with methanol and the eluate was lyophilized (Process I). Alternatively, the pectin was precipitated by adding the crude extract to ethanol. After removal of the organic solvent, the phenolic compounds were obtained from the aqueous phase of the precipitation bath using the adsorbent resin as described before (Process II). While in total, 129.4 mg/g polyphenols were detected in the lyophilizate obtained from Process I, only 71.0 mg/g dm could be recovered from Process II. The profiles of the polyphenols were almost identical, revealing that during pectin precipitation preferential adsorption of polyphenolic compounds to the pectin may be excluded. Besides the characterization of the pectins and the phenolic compounds, investigations into the influence of the drying temperature on the polyphenolic content of the peels were carried out, indicating a significant loss of flavonol glycosides depending on heat exposure. On the other hand, some xanthone glycosides were formed during the drying process. Furthermore, antioxidative capacities of the lyophilized eluates were investigated using the DPPH, TEAC and FRAP assays. The antioxidative capacity of the extracts exceeded that of mangiferin and quercetin 3-O-glucoside, respectively, thus demonstrating mango peels to be a suitable source of health-beneficial compounds. The lyophilizates obtained from Process I showed higher antioxidative capacities in all three assays. These findings indicate a correlation between the amount of phenolic compounds and the antioxidative capacity.
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Study On Mango Leaf and Mangiferin Vascular effects of the Mangifera indica L. extract (Vimang)
Beltrn AE, Alvarez Y, Xavier FE, Hernanz R, Rodriguez J, Nez AJ, Alonso MJ, Salaices M. Depto. de Farmacologa y Teraputica, Facultad de Medicina, Universidad Autnoma de Madrid, Madrid, Spain
The effects of the Mangiferia indica L. (Vimang) extract, and mangiferin (a C-glucosylxanthone of Vimang) on the inducible isoforms of cyclooxygenase (cyclooxygenase-2) and nitric oxide synthase (iNOS) expression and on vasoconstrictor responses were investigated in vascular smooth muscle cells and mesenteric resistance arteries, respectively, from Wistar Kyoto (WKY) and spontaneously hypertensive (SHR) rats. Vimang (0.5-0.1 mg/ml) and mangiferin (0.025 mg/ml) inhibited the interleukin-1beta (1 ng/ml)-induced iNOS expression more in SHR than in WKY, and cyclooxygenase-2 expression more in WKY than in SHR. Vimang (0.25-1 mg/ml) reduced noradrenaline (0.1-30M)- and U46619 (1nM-30M)but not KCl (15-70 mM)-induced contractions. Mangiferin (0.05 mg/ml) did not affect
noradrenaline-induced contraction. In conclusion, the antiinflammatory action of Vimang would be related with the inhibition of iNOS and cyclooxygenase-2 expression, but not with its effect on vasoconstrictor responses. Alterations in the regulation of both enzymes in hypertension would explain the differences observed in the Vimang effect.
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Study On Mango Leaf and Mangiferin Vimang (Mangifera indica L. extract) induces permeability transition in isolated mitochondria, closely reproducing the effect of mangiferin, Vimang's main component
Pardo-Andreu GL, Dorta DJ, Delgado R, Cavalheiro RA, Santos AC, Vercesi AE, Curti C. Departamento de Fsica e Qumica, Faculdade de Cincias Farmacuticas de RibeiroPreto, Universidade de So Paulo, Ribeiro Preto, SP, Brazil
Mitochondrial permeability transition (MPT) is a Ca2+-dependent, cyclosporin A (CsA)-sensitive, non-selective inner membrane permeabilization process. It is often associated with apoptotic cell death, and is induced by a wide range of agents or conditions, usually involving reactive oxygen species (ROS). In this study, we demonstrated that Mangifera indica L. extract (Vimang), in the presence of 20 M Ca2+, induces MPT in isolated rat liver mitochondria, assessed as CsA-sensitive mitochondrial swelling, closely reproducing the same effect of mangiferin, the main component of the extract, as well as MPT-linked processes like oxidation of membrane protein thiols, mitochondrial membrane potential dissipation and Ca2+ release from organelles. The flavonoid catechin, the second main component of Vimang, also induces MPT, although to a lesser extent; the minor, but still representative Vimang extract components, gallic and benzoic acids, show respectively, low and high MPT inducing abilities. Nevertheless, following exposure to H2O2/horseradish peroxidase, the visible spectra of these compounds does not present the same changes previously reported for mangiferin. It is concluded that Vimang-induced MPT closely reproduces mangiferin effects, and proposed that this xanthone is the main agent responsible for the extract's MPT inducing ability, by the action on mitochondrial membrane protein thiols of products arising as a consequence of the mangiferin's antioxidant activity. While this effect would oppose the beneficial effect of Vimang's antioxidant activity, it could nevertheless benefit cells exposed to over-production of ROS as occurring in cancer cells, in which triggering of MPT-mediated apoptosis may represent an important defense mechanism to their host.
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Xanthone derivatives: new insights in biological activities

Pinto MM, Sousa ME, Nascimento MS Centro de Estudos de Qumica Orgnica, Fitoqumica e Farmacologia da Universidade do Porto, Laboratrios de Qumica Organica e Microbiologia, Faculdade de Farmcia, Porto, Portugal
Xanthones or 9H-xanthen-9-ones (dibenzo--pirone) comprise an important class of oxygenated heterocycles whose role is well-known in Medicinal Chemistry. The biological activities of this class of compounds are associated with their tricyclic scaffold but vary depending on the nature and/or position of the different substituents. In this review, an array of biological/pharmacological effects is presented for both natural and synthetic xanthone derivatives, with an emphasis on some significant studies on structure-activity relationships. The antitumor activity of some xanthones as well as the related targets, particularly PKC modulation studies, is also discussed in detail. Examples of the "hit" compounds involved in cancer therapy, namely DMXAA, psorospermin, mangiferin, norathyriol, mangostins, and AH6809, a prostanoid receptor antagonist, are also mentioned. Finally, a historical perspective of these xanthonic derivatives, their relevance as therapeutic agents and/or their uses as pharmacological tools and as extract components in folk medicine are also highlighted.
Xanthone glycosides from herbs of Polygala hongkongensis Hemsl and their antioxidant activities
Wu JF, Chen SB, Gao JC, Song HL, Wu LJ, Chen SL, Tu PF. Shenyang Pharmaceutical University, Shenyang,P.R.China
Two new xanthone O-glycosides, polyhongkongenosides A and B, together with four known xanthone glycosides, were isolated from the herbs of Polygala hongkongensis Hemsl. Their structures were elucidated on the basis of UV, IR, NMR, and MS spectral data. The antioxidant in vitro activities of 1-6 were determined by the scavenging activities against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals and hydroxyl radicals and their reductive activities to Fe3+. Mangiferin, one of the four known xanthone glycosides, showed potential scavenging effect on DPPH and hydroxy radicals and reductive activity to Fe3+ with IC50 values of 4.7, 13.9, 23.7 M, respectively.
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Study On Mango Leaf and Mangiferin Pharmacokinetics of mangiferin in rat plasma after oral administration of a single dose of Suanzaoren decoction
Yujuan Li, Kaishun Bi School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, P.R.China
This research studied the pharmacokinetics of mangiferin in rats after oral administration of a single dose of Suanzaoren decoction. An HPLC method was established using puerain as internal standard. Plasma samples were deproteinized with acetonitrile-acetic acid (9:1), followed by evaporation of the acetonitrile to dryness. The resultant residue was then dissolved in mobile phase and HPLC separation was achieved on a Hypersil C18 (200 mm 4.6 mm D, 5m ) column at room temperature. The mobile phase consisted of acetonitrile-water (12:88) with 1% acetic acid and 1% tetrahydrofuran at a flow rate of 0.7 mLmin-1. The UV detection wavelength was set at 320 nm. The calibration curve was shown to be linear over the range from 0.536 to 26.8 gmL
1
( r2
0.995). Mean recovery was determined as 92.7%.
Within-day and between-day precisions were less than 9.1% RSD. The limit of quantitation (LOQ) was 0.536 gmL-1. The maximum plasma concentration (Cmax), the time to reach peak concentration (Tmax ) and the apparent elimination half-life ( T1 /2 ) were (10.5 2.2) gmL-1 , ( 5.8 0.4) h and ( 5.0 0.3) h, respectively. We could see the validated HPLC method developed has been applied to take a limited view of pharmacokinetics profile of mangiferin in rat plasma after having orally taken a single dose of Suanzaoren decoction.
Synthesis of mangiferin derivates and study their potent PTP1B inhibitory activity
Honggang Hu1.2, Mingjuan Wang2, Qingjie Zhao2, Shichong Yu2, Chaomei Liu2, Qiuye Wu2
1 2
Department of Chemistry, College of Science, Wayne State University, United States Department of Organic Chemistry,School of Pharmacy, Second Military University, Shanghai, P.R.China
Protein tyrosine phosphatase 1B(PTP1B)has received considerable attention from the drug industry as a potential treatment for diabetes mellitus.Mangiferin has been reported to possess significant activity.Based on the previous study,eight new mangiferin derivates were synthesized and evaluated for their PTP1B inhibitory activity. Some of them display good inhibitory activity on PTP1B.
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Study On Mango Leaf and Mangiferin Pharmacokinetics of mangiferin in rat plasma after oral administration of a single dose of Suanzaoren decoction
Yujuan Li, Kaishun Bi School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, P.R.China
This research studied the pharmacokinetics of mangiferin in rats after oral administration of a single dose of Suanzaoren decoction. An HPLC method was established using puerain as internal standard. Plasma samples were deproteinized with acetonitrile-acetic acid (9:1), followed by evaporation of the acetonitrile to dryness. The resultant residue was then dissolved in mobile phase and HPLC separation was achieved on a Hypersil C18 ( 200 mm 4.6 mm D, 5 m ) column at room temperature. The mobile phase consisted of acetonitrile-water (12:88) with 1% acetic acid and 1% tetrahydrofuran at a flow rate of 0.7 mLmin-1. The UV detection wavelength was set at 320 nm. The calibration curve was shown to be linear over the range from 0.536 to 26.8 gmL
1
( r2
0.995). Mean recovery was determined as 92.7%.
Within-day and between-day precisions were less than 9.1% RSD. The limit of quantitation (LOQ) was 0.536 gmL-1. The maximum plasma concentration (Cmax), the time to reach peak concentration (Tmax ) and the apparent elimination half-life (T1/2 ) were (10.52.2) gmL-1 , (5.80.4) h and (5.0 0.3) h, respectively. We could see the validated HPLC method developed has been applied to take a limited view of pharmacokinetics profile of mangiferin in rat plasma after having orally taken a single dose of Suanzaoren decoction.
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Study On Mango Leaf and Mangiferin Effect of Mangifer in on telomerase activity and cell cycle in K562 cells
Zhigang Peng 1, Jun Luo 1, Yongrong Lai 2, Shanjun Song 2
1
Department of Hemotology, The First Affiliated Hospital, Guangxi Medical University, Nanning,
P.R.China
2
Department of Hemotology, Union Hospital, Tongji Medical College, Huazhong University of Science
and Technology, Wuhan, P.R.China
This study wanted to investigate the effects of mangiferin on telomerase activity and cell cycles in K562 cells lines, and further to study the molecular mechanism of its anti-leukemia. Polymerase chain reaction enzyme linked immunoassay (PCR-ELISA) was used to assay telomerase activity of K562 cells. The change of cell cycle in K562 cells treated with mangiferin was performed by the folw cytometric. The level of telomerase activity of K562 cells decreased as the cells treated by mangiferin, in a tim- and concentration-dependentmanner. After treatment with mangiferin for 24 hours, the cell cycle distribution changed, the percentage G2/M stage cell increased in a concentration dependent manner, indicating G2/M phase arrest. The results showed that Mangiferin could defectively inhibit tolomerease activity. Arresting cell cycle may be the machnisams.
Effect of Mangiferin on the Content of PGE2 in Two Different Inflammation Models

Jiagang Deng, Erwei Hao, Zuowen Zheng, Ke Yang, Li Yan Guangxi Traditional Chinese Medical University, Nanning, P.R.China
This research studied the effect of mangiferin on the content of PGE2 in two different inflammation models and elucidated its mechanisms of anti-inflammation. Mice air-pouch acute inflammatory model induced by carrageenan and rabbit systemic inflammatory model induced by endotoxin were adopted and PGE2 production in two different inflammation models were respectively detected by ultraviolet spectrophotography and radioimmunoassay. The results showed that the mangiferin can obviously inhibit the content of PGE2 in inflammatory exudates of mice back gasbag, and in hypothalamic of Systemic Inflammatory rabbit model. These results suggest that the mechanism of anti-inflammatory effect of Mangiferin might be associated with its inhibitory effect on PGE2.
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Study On Mango Leaf and Mangiferin The Effect of Mangifer in on Telomerase Activity and Apoptosis in Leukem ic K562 Cells
Peng Cheng1, Zhigang Peng1, Jie Yang1 , Shanjun Song2
1
Department of Hemotology, The First Affiliated Hosp ital, Guangxi Medical University, Nanning, P.R.China
Department of Hemotology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, P.R.China
The research investigated the effects of mangiferin on telomerase activity and apoptosis in K562 cells lines and studied the molecular mechanism of its antileukemic. Cell apoptosis was observed by light microscopy and transmission electron microscopy, and the expression of Fas was measured by flow cytometry. Polymerase chain reaction enzyme linked immunoassay (PCR-ELISA) was used to assay telomerase activity of K562 cells. The results showed that Mangiferin could inhibit telomerase activity of K562 cells in a time-and-concentration-dependent manner. Meanwhile, it could induce apoptosis obviously and up-regulate the levels of Fas in K562 cells. For this, we may see Mangiferin can inhibit telomerase activity of K562 cells, and the mechanism of effect is maybe related to inducing apoptosis and the expression of Fas protein.
The Antitussive and Expectorant Effects of Mangifera Leaves Extract

Guofeng Wei, Zuliang Huang,Youcheng He Department of Applied Chemistry, You jiang Medical College for Nationalities, Bose, Guangxi, R.P.China
This research investigated the antitussive and expectorant effect of mango leaves extract on mice with cough. The extract was separated from mango leaves using distilled water and ethanol as solvents. An antitussive and expectorant test was performed on mice with cough induced by ammonia water. The mango leaves extract was effective on mice with cough, in comparison with the control group, there was statistically significant difference(P<0.05 or 0.01).Mango leaves extract has significant anti-titussive and expectorant efficacy.
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Study On Mango Leaf and Mangiferin Preliminary Studies on the Mode of Action of Mangifer in against Phytophthora infestans
Fengping Song1, Shutong Wang 1, Tongle Hu1, Jianjian Wei2, Keqiang Cao1
1 2
College of Plant Protection, Agricultural University of Hebei ,Hebei P.R.China Beijing TEPEC Science Corporation, Beijing, P.R.China
Mangiferin is an active ingredient of traditional Chinese medicinal herb Anemarrhena asphodeloides. Bioassay tests in vitro showed that mangiferin could inhibit the mycelium growth of Phytophthora infestans in tensively, with EC50 value 32.65g/mL, but it has low inhibitory effect on the zoospore release of Phytophthora infestans with EC50 value 100.97g/mL.Coomassie brilliant blue G-250 colorimetric method was used to test the protein variations of Phytophthora infestans mycelia. The result indicated that mangiferin inhibited the protein biosynthes is of Phytophthora infestans. It could influence the permeability of the cell membrane of Phytophthora infestans at higher concentration, but the effect was not significant at the lower concentration 12.5g/mL.Dissolved oxygen content in zoospores suspension was used to test the respiratory inhibition effect of mangiferin on Phytophthora infestans. The results revealed that mangiferin had significant inhibition effect on metabolic path ways such as glycolytic pathway (EMP), tricarboxy licacid cycle (TCA) and hexose monophosphate pathway (HMP), and the inhibition effect on HMP was the most significant.
Experimental Study on Anti-bacterial, Anti-inflammatory and Analgesic Activities of the Mixture of Mangiferin and Berberine
Xuejian Li, Jiagang Deng, Zhenlin Qin, Dongyan Liao, Xiaoni Lu Guangxi Traditional Chinese Medical University, Nanning, P.R.China
The aim of the present study is to study the anti-bacterial, anti-inflammatory and analgesic activities of the mixture of mangiferin and berberine (MMB). Anti-bacterial activity was test in tubes in which the liquid was diluted to double volume everytime; anti-inflammatory activity was tested by a rat model with uterine inflammation; analgesic activity was tested by observing the writhing response caused by acetic acid. MMB had inhibitory effect on a variety of bacteria, on rat uterine inflammation and could reduce the writhing times caused by acetic acid. MMB has anti-bacterial, anti-inflammatory and analgesic activities.
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Study On Mango Leaf and Mangiferin An Experimental Study of Anti-stress Effects of Mangiferin in Mice
Jianquan Wei, Zimin Zheng, Yong Pan, Xiaolin Xu, Qiang Xue, Binxue Huang, Shu Lai, Zengqiong Huang Department of Pharmacology of Youjiang Medical College for Nationalities, Guangxi, P.R.China
The study observed the anti-stress effects of Mangiferin in mice. One hour after different concentrations and doses of Mangiferin and distilled water were administered using Gavage method,mice were divided into groups for anti-stress tests like loaded swimming, resistance to lack of oxygen and low temperature to observe changes of superoxide dismutase (SOD) vitality and concentrations of Malondialdehyde (MDA). The results is that the survival time of the tested mice were longer than the control groups(P<0.01)with sera SOD vitality increased and MDA concentration decreased(P<0.01 or 0.05). Mangiferin had certain anti-stress actions.
Preliminary study on effects of mangiferin on immunologic function in mice

Huaizhou Qin, Liang Wang, Zhenwei Zhao, Lihong Li, Jianping Deng, Youzhi Qu, Hangyu Shi, Li Gao, Guodoug Gao Department of Neurosurgery,Tangdu Hospital, Fourth Military Medical University; Institutefor Functional Neurosurgery of PLA, Shanxi, .RP.China
The research investigated the effects of mangiferin purified from leaves of Mangifera indica L.on immunologic function in mice. The olig-immunity model was made with hydrocortisone. The indexes of immune organs were calculated. The phagocytosis of mononuclear macrophage was determined with carbon particle clearance test. The spectrophotography was used todetect the levels of serum hemolysis IgG and IgM. The cell proliferation was measured by MTT assay. The mangiferin of 50 and 100 mg/kg significantly increased the phagocytic function, and recovered the indexes of the spleen and thymus in immunosuppressive mice caused by hydrocortisone. It also remarkably increased the levels of serum hemolysis IgG and IgM. The mangiferin could increase the proliferation of mouse spleen cells and macrophage. These results suggest that the mangiferin could enhance the non-specific immunity and humoral immunity in immunosuppressive mice.
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Study On Mango Leaf and Mangiferin Effects of mangiferin of TNF- and MPO in rats with myocardial ischemia reperfusion injury
Chao Wang, Guoxian Wang Dept of pharmacology, Jinzhou Medical College, Jinzhou, RP.China
The study investigated the effect of Mangiferin on attenuating TNF-a and MPO in rats with myocardial ischemia reperfusion (MIR) injury. Fifty SD rats were randomly divided into five groups sham, model, Mangiferin groups of 10mg/kg, 20mg/kg, 40mg/kg, sham group with equal volume of saline intraperitoneal injected while was given all the procedures except ligation. mangiferin groups were injected with corresponding concentrations of mangiferin intraperitoneal injucted for 21 days.one hour after the last administration for the experiment. Myocardial ischemia reperfusion models were obtained by ligated left anterior descending coronary artery 40 minutes and followed by 120 minutes reperfusion. Detected Myocardial Tumor necrosis factor-a (TNF-a) with RIA; Detected Infiltration of polymorpho nuclear neutrophils (PMN) in myocardium with Myeloperoxidase (MPO) observe pathologic changes of myocardium with HE staining, observed ultrastructure of myocardium with TEM. Compared with the ,model group, mangiferin group TNF-a, MPO were significantly lower, mangiferin group could improve myocardial pathologic and ultrastructure and pathologic. Mangiferin can protect MIR. The myocardial protective mechanism of may be realized by Alleviate Infiltration of polym-errphonuclear ncutrophils (PMN) in myocardium, inhibit inflammatory factor liberation relate.
Experimental Study on Hypoglycemic Effect of the Mixture of Mangiferin and Berberine

Xuejian Li, Jiagang Deng, Zhenlin Qin, Ning Liang, Dongyan Liao Guangxi Traditional Chinese Medical University, Nanning, P.R.China
The research studied that the hypoglycemic effect of the mixture of Mangiferin and Berberine (MMB). Experimental observation was performed by using various diabetic mice induced by Alloxan,
adrenalin and glucose. MMB could significantly lower the blood glucose levels in Alloxan model mice, adrenalin model mice and hyperglycemic mice, while had no hypoglycemic effect on the normal mice .MMB has hypoglycemic effect in diabetic model mice.
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Study On Mango Leaf and Mangiferin Effect of mangiferin on myocardial ischemia induced by isoproteronol in mice
Jianquan Wei, Zimin Zheng, Yong Pan, Ying Luo, Jian Huang, Shu Lai Department of Pharmacology of Youjiang Medical College for Nationalities, Guangxi, P.R.China
The study evaluated the protective effect of mangiferin (MAN) on myocardial ischemia induced by isoproteronol (ISO) in mice. Acute myocardial ischemic model induced by intraperitoneal administration of ISO in mice was adopted to observe the effects of MAN on the abnormality in lead on ECG, the myocardial contents of SOD and MDA, the serum levels of LDH and CK. MAN could ameliorate the abnormal changes of ECG, could increase the myocardial activity of SOD and could decrease the myocardial contents of MDA, the serum levels of CK and LDH. MAN has protective effect on myocardial ischemia induced by ISO in mice.
Apoptotic mechanism of leukemic K562 cells induced by mangiferin

Zhigang Peng 1, Jun Luo 1, Yongrong Lai 1, Shanjun Song 2
1
Department of Hemotology, The First Affiliated Hospital, Guangxi Medical University, Nanning, P.R.China
This research investigated the apoptosis mechanism of K562 cell lines induced by mangiferin. The mRNA expression levels of apoptosis-related genes including bcl-2, bax, survivin of K562 cells treated by mangiferin (25-200mol /L) for 24, 48, 72 and 96h were determined by RT-PCR; the BCR /ABL protein P210 level was detected by Western blotting. Mangiferin up-regulated bax gene of K562 cells significantly and down-regulated bcl-2 gene slightly, resulting in an enhancement of the ratio of bax bcl-2.Mangiferin down-regulated the expression levels of P210 in K562 cells in a time-and concentration dependent manner and so is the expression level of surviving mRNA in K562 cells. The mechanism of mangiferin-induced apoptosis in K562 leukemic cells might be involved in up-regulating the gene expression of bax and down-regulating the mRNA expression of BCR /ABL protein P210, bcl-2,and surviving.
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Study On Mango Leaf and Mangiferin Inhibitory effect of mangiferin on duck hepatitisB virus (DHBV) DNA in vivo
Jiagang Deng, Ke Yang, Zuowen Zheng, Xiaoxue Zhou Guangxi Traditional Chinese Medical University, Nanning, P.R.China
This research investigated the inhibitory effect of Mangiferin on duck hepatitis B virus DNA in vivo.1-day-old peking ducks were administered orally with Mangiferin, either of 50mg/kg, or 100 mg/kg, or 200mg/kg for twice a day, 10 days on end after intravenous administration of DHBV. Blood collection was performed before the treatment, which was also applied to the 5th, 10th day and 3 days after the treatment. Dynamic changes of DHBV DNA were observed by dot-blot hybridization test on the isolated serum. Mangiferin markedly inhabited the DHBV DNA when the dose of which was 100mg/kg or 200mg/kg. There was no obvious rebound phenomenon shown after the treatment. Mangiferin has favorable inhabiting effect on DHBV DNA. Rebound phenomenon rarely occurs after administration of Mangiferin.
Comparison Tests of the Efficacy of Mango Leaf Decoction, Demangiferin Mango Leaf Decoction and Mangiferin Anti-tussive and Expectorant Drugs
Naiqiu Wei, Jiagang Deng, Hanmei Xian, Jinluan Wei Teaching Office of Chinese Herbology, Guangxi College of Traditional Chinese Medicine, Nanning, Guangxi, P.R.China
This research compared the pharmacological action of mango leaf decoction,demangiferin mango leaf decoction and mangiferin antitussive and expectorant drugs. The methods of thick ammonia water-inducing cough, sulphur dioxide-inducing cough and phenolsulfon phthalein excretion test were used respectively to observe the anti-tussive and expectorant effect of three drugs. All the high dose and medium dose of three drugs could remarkably resist the cough rates caused by thick ammonia water and sulphur dioxide and delay the cough latent period. All the three drugs have expectorant action, esp. the high dose of mangiferin. All the three drugs have anti-tussive and expectorant action, among which the high dose of mangiferin group has the best effect.
113
Study On Mango Leaf and Mangiferin Study on Antibacterial Action of Extract of Leaves of Mangifera indica in Vitro
Xiuping Liu, Weizhe Jiang, Xingzhen Huang, Zengqiong Huang, Min Huang Center of Drug Research and Development, the First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, R.P.China
This research observed the antibacterial action of extracts of the leaves of Mangifera indica on common clinical bacterials in vitro with the two-fold agar dilution method. The extracts of Mangifera indica showed the strong inhibitory effects on common clinical pathogens such as P.aeruginosa, E.coli, K.peneumoniae, Acinetobacter baumannii, S.aureus, S.epidermidis. MIC of the extracts of the leaves of Mangifera indica was between 0.107-1.320 mg/ml. The activity of inhibition was affected by inoculative quantity and pH value of dilution, MIC was decreased when inoculative quantity was increased or the pH value of dilution was decreased. The extracts of the leaves of Mangifera indica has strong inhibition on bacteria in vitro. The results provide a basis for clinical application of mangiferin.
Inhibiting effect in vitro of extract of Mangifera indica L.leaf on some pathogenic bacteria and NDV replication
Zhongsheng Xia, Chunhua Han, Guoying He, Tingchong Tang, Liping Wei College of Animal Science and Technology,Guangxi University,Nanning ,R.P.China
An inhibiting-bacteria test was done about effects of extract of mangifera indica L.leaf on some pathogenic bacteria of domestic animal and poultry by using paper method, and an inhibiting-NDV experiment by extract of mangifera indica L. leaf with chicken embryos infected artificially was carried out. The result of inhibiting-bacteria test showed that extract of mangifera indica L. leaf had strong inhibiting effect on E scherichia coli, Pasteurella multocid a, Salmonella pullorum. Erysip eloth rix rhusiopathiae, Staphylococcus aureus, Riemerella anatipestifer, Strep tococcisuis. The results of inhibiting-NDV showed that at 48h the chicken embryos protective rate of NDV control group and 2.500 g/ mL+NDV group were 10.00% and 63.64%, respectively. There was significant difference between groups (P<0.05).The protective rate of 1.250g /mL+NDV group,0.625g/ mL+NDV group and 0.313 g /mL+NDV group were 72.72%, 70.00%, 69.23%, respectively.There were extremely significant differences (P<0.01) compared with NDV control. It was suggested that extract of mangifera indica L. leaf had inhibiting-NDV effects.
114
Study On Mango Leaf and Mangiferin The Ultramicro-structure Change of Lipid Superoxided Rat Brain Tissue and Protect of Mangiferin on the Tissues
Huayi Huang, Ming Zhong, Chaozan Nong, Shiyuan Zhao, Gang Meng Guangxi Nationalities Hospital,34 East Mingxiu lu,Nanning,Guangxi,R.P.China
This research investigated the ultramicro-structure change of lipid superoxided rat brain tissue and protect of mangiferin on the tissues, we established a lipid superoxided model for rat using chemical substance-alloxan for our study. Rats were divided into 5 groups: Group A: the animals were gastrogavaged 10ml/day of 5% starch solution for 21 days .Group B:30 mg/kg/day of mangiferin for 21 days.Group C:15 mg /kg/day of mangiferin for 21 days. Group D:75 mg /kg/day of VitE for 21 days. Group E:10ml/day of saline for 21 days, and Groups B to E were also intraperitoneal injected 75 mg/kg/day of alloxan from day 16 to day 21.The animals were then executed for perfuse fixation, and ultramicrostructure change in caudate nucleus of brain tissue were observed. The forms of organelles in neurons of Group A were normal, N v:(1.2330.102) /m 3.,w hile it showed that Group Bs were roughly normal, N v:(1.3420.098) /m 3.The mitocondrial cristae in neurons of Group C were reduced, the width between crista increased. N v reduced (0.83760.075).The organelles in neurons of Group D were roughly normal, but the width between crista increased,N v:(1.2210.092).Organelles in neurons of Group E were abnormal, cristae reduced, the width between crista increased significantly, and the N v also reduced significantly (0.82620.058). Mangiferin can scavenge lipids superoxides from brain tissue,and protect the normal function of neuron.
115
Study On Mango Leaf and Mangiferin Inhibitory effect of mangiferin on the proliferation of K562 leukemia cells
Zhigang Peng1, Jun Luo1, Linghu Xia1, Shanjun Song2, Yan Chen2 1 Department of Hemotology, The First Affiliated Hospital, Guangxi Medical University, Nanning, P.R.China 2 Department of Hemotology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, P.R.China
The MTT assay, colony formation assays and growth curves methods were adopted to study the effects of growth inhibition of mangiferin on K562 leukemia cells in vitro. All of the three methods indicated that different concentrations of mangiferin could inhibit the K562 cells proliferation in different times and the inhibition effect were dependent on time and concentration; and the MTT assays showed that the inhibition of K562 cells was 31.99%, 50.74%, 60.24%, 62.65% and 70.41% respectively with (25-200mol /L) mangiferin treatment for 72 hours, the inhibition of K562 cells was 48.19%, 68.88%, 74.38%, 76.93% and 83.50% respectively with 200mol/L mangiferin treatment for 24, 48, 72 and 96h.This conclusions showed that the proliferation of K562 cells was markedly inhibited by mangiferin in a dose-dependent and time-dependent manner.
116
Effect of mangiferin on the expression of -cateninand p120ctn in hepatic tissues of rats with liver cancer
Chaozan Nong, Lingxiao Guo, Huayi Huang Central Laboratory of Clinical Medicine Institute of Guangxi Hospital f or Nationalities, Biology Laboratory of Tumor Molecular and Cellula, Nanning, P.R.China
The methods adopted cells of liver cancer of Walker 256 and Wistar rats for liver cancer model and observed expressions of -catenin and p120ctn in hepatic tissues by immunohistochemical techniques. The results indicated Group A: The expressions of the above cells were of cytomembrane while no expressions for kytoplasm; Group B: Expressions of both cytomembrane and kytoplasm or decrease of that of kytoplasm were detected while some cytomembranes lacked expression; Group C:Obvious expressions of cytomembrane and certain kytoplasms were detected; Group D: Obvious expressions of both cytomembrane and kytoplasm with cytomembrane dominating, some cells showed no expressions; Group E: Expressions of both cytomembrane and kytoplasm appeared with kytoplasm dominating, decrease of expressions and discontinuity occurred. The conclusion showed that mangiferin was definite stabilization in the normal expressions of cell signal transduction prompting that it possessed potential anti-tumor substances in the cell adhesion and channels of signal transduction of mediate.
Effects of enzyme and morphological change of mangiferin on experimental liver damage in rats
Hailong Cheng,Yuhua Li,Qingya Bian Yancheng Health School, Yancheng, Jiangsu, P.R.China
The mangiferin was investigated on the hepatoprotective effects in three kinds of models of experimentally-induced liver injuries in rats. The models were induced by acetaminophen, carbon tetrachloride, and D-GalN. The results showed that mangiferin caused significant reduction of the elevated enzyme levels of serum glutamate oxaloacetate transminase, serum glutamate pyruvate transminase in model rats, and alleviated the pathological damages in livers of the model rats. These findings were suggestive of the potent hepatoprotective effects of mangiferin.
117
Study On Mango Leaf and Mangiferin The proliferation inhibition effect and apoptosis induction of mangiferin on BEL-7404 human hepatocellular carcinoma cell
Huayi Huang ,Chaozan Nong, Lingxiao Guo, Gang Guo, Xiliang Zha Department of Experimental Center, Guangxi Nationalities Hospital, Nanning, P.R.China
The MTT assay was used to determine the effect of Mangiferin on cell proliferation, microscopy method was used to observe cytotoxicity of Mangiferin, and flow cytometry method was used to determine apoptosis induction and cell cycle proliferation blocking effects of Mangiferin. The results indicated that the cytotoxicity effects were observed in various concentrations of Mangiferin and at different exposure times. The effects were enhanced as the Mangiferin concentration increased and exposure time prolonged. The apoptosis effect was also enhanced, and cell cycle arrested at G2/M phase. All the above effects became significant when cells exposed to the concentration from 20mol/L to 200mol/L of Mangiferin at 24h.The conclusion show that Mangiferin is cytotoxic, and may induce apoptosis. Mangiferin has cell cycle blocking effects on hepatocellular carcinoma cells. Its pharmaceutical potentials is worth for further investigating.
Effect of mangiferin on P120ctn phosphorylation and hepatocellular carcinoma cell biology

Shaoyun Nong, Chaozan Nong, Lili Pan, Weisheng He, Huayi Huang Guangxi Nationalities Hospital, Nanning, P.R.China
In this study, Human hepatocellular carcinoma cell line were sitimulated with EGF to make P120ctn tyrosine phosphorylation, tyrosine phosphorylation of P120ctn was detected by immunoprecipitation and immunoblotting methods and investigated the Mangiferin infection in the carcinoma cell. The result indicated: Phosphorylated tyrosine of P120ctn after treatment with EGF was detected; while Mangiferin reduced tyrosine phosphorylation of 120ctn phosphorylated P120ctn, cell adhesion aecreased and migration ability increased, Mangiferin had reversed these phenomenons in some scale. The conclusion show that: P120ctn play an important role in hepatocellular carcinoma cell adhesion and the tyrosine phosphorylation of 120ctn was involved in Hepatocellular carcinoma cell malignant behavior enhancement. Mangiferin restrain heap-tocellular cacinoma P120ctn tyrosine phosphorylation.
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Study On Mango Leaf and Mangiferin Experimental study on effect of mangiferin delaying caducity
Peng Zhao, Li He, Junfeng Yang, Bin Li, Rongzhen Liu, Jian Liang, Fengwen Li, Chaopei Huang Guangxi Zhuang Autonomous Region Center for Disease Prevention and Control, Nanning, P.R .China
Two methods were used to study the effect of mangiferin delaying caducity. (1)The experimented aged rats were continually given the tested sample 100 days, with dose of 5.00,2.50,1.25mg/kg.bw separately, then contents of LPO and SOD in their sera or tissues being tested.(2)The experimented drosophila melanogasters were fed in culture media which were mixed into the tested sample with dose 1.50,0.30,0.06mg/100ml separately, their life time being observed. The results showed that: (1) the contents of LPO and lipofuscinin serum and brain tissue of rats which were given sample were less than those of controls, but their SOD levels were higher than those of controls. (2)The average life time and the most long life time of drosophila melanogasters which were given sample were longer than those of controls. The conclusion investigate that mangiferin has the effect of delaying caducity.
Effects of Mangifer in on induction of apoptosis and in tracellular Ca2+ concentration in Nasopharyngeal Carcinoma CNE2 Cells
Xiaochun Liu1, Jiao Lan1, L ili P an1, Zhaozan Nong2
1 2
The Peoples Hospital of Guangxi, Nanning, PR China Region Hospital of Guangxi, Nanning, P.R.China
This experiment was armed to study the effects of mangiferin on cell apoptosis and intracellular Ca2+ concentration in CNE2 cells of nasopharyngeal carcinoma in vitro. The method was used to measure the different levels of mangiferin for 24 hours, 48 hours and 72 hours by flow cytometry, later these items showed the change of the apoptosis ratio and IECa2+ content in CNE2 cells. The results showed
that:(1)The each level of mangiferin could induce the death of CNE2 cells for 24 hours, 48 hours and 72 hours, his phenomenon was dependent on effect time and levels of mangiferin.(2)The IECa2+ content in CNE2 cells had also increased obviously by mangiferin treatment for 24 hours, 48 hours and 72 hours, meanwhile the time and levels of mangiferin varied directly with the IECa2+content.The conclusion investigate that mangiferin can induce obviously the death of CNE2 cells, and the increased content of IECa2+ in CNE2 cells should play an important role to induce the death of ones.
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Study On Mango Leaf and Mangiferin CML cell line K562 cell apoptosis induced by Mangiferin
Zhigang Peng1, Jun Luo1, Linghui Xia2, Yan Chen2, Sanjun Song2
1
Department of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science
and Technology, Wuhan, P.R.China

2
Department of Hematology, the First Affiliated Hospital, Guangxi Medical University, Nanning,
P.R.China
This study was aimed to investigate the effects and the mechanism of mangiferin on chronic myeloid leukemia cell lines K562 cells in vitro. The antiproliferation effects of mangiferin on K562 leukemia cells were tested by tetrazolium salt (MTT)method; the apoptosis induced by mangiferin on K562 cell line was explored by means of cell morphology, DNA gel electrophoresis and flow cytometry. The changes in bcr/abl gene expression were detected by using reverse transcriptase (RT)-PCR. The results showed that five different concentrations of mangiferin (25-200mol/L) dose-dependently and time-dependently inhibited the proliferation of K562 cells, and induced apoptosis in K562 cell line. RT-PCR revealed that bcr/abl gene expression was down-regulated when K562 cells had been treated with different concentrations of mangiferin. In conclusion, mangiferin remarkably inhibits t he proliferation of K562 leukemia cells in vitro, and induces apoptosis in K562 cell line probably through down-regulation of bcr/abl gene expression.
Pharmacodynamic studie s on Mangiferin

Jiagang Deng, Zuowen Zheng, Chunhui Zeng Guangxi Traditional Chinese Medical University, Nanning, P.R.China
The asthma test caused by histamine and acetylcholine in guinea pig, cough test aqua ammoniae-induced, permeation of capillary vessel, emission of phenol red in mice trachea were examined to investigate the pharmacodynamics of mangiferin. The results showed that mangiferin prolonged markly the incubation period of asthma and cough and decreased cough times, inhibited the permeation of capillary vessel caused by acetic acid and increased the emission of phenol red.
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Experimental study on the pharmacology of Mangiferin monosodium salt

Yefei Yuan1,2, Jiagang Deng 1, Xin Yu 2, Ying He2
1 2
Guangxi Traditional Chinese Medical University, Nanning, P.R.China Luzhou Medical College, Luzhou, P.R.China
This study was aimed to investigate the effect of mangiferin monosodium salt on respiratory system. The methods of sulfur dioxide induced cough, phenolsulfonphthalein excretion test, histamine phosphate and acetylcholine induced asthma, and dimethy benzene induced swelling ear were adopted to observe the antiinflammatory antitussive, expectorant effects by abdominal injection . The results showed that mangiferin monosodium salt was able to prolong cough latent period and decrease the frequency of cough significantly in mice resulted from sulfur dioxide, significantly increase phenolsulfonphthalein excretion quantity of tracheal in mice, prolong asthma latent period in mice induced by histamine phosphate and acetylcholine, and inhibit obviously the ear edema induced by dimethyl benzene in mice. In conclusion, mangiferin monosodium salt has antitussive, expectorant, antiasthmatic and anti-inflammatory actions.
Preparation
of
mengiferin
monosodium
salt
and
comparison
in
pharmacological effects with mengiferin

Jiagang Deng1, Yefei Yuan 2
1 2
Guangxi Traditional Chinese Medical University, Nanning, P.R.China Luzhou Medical College, Luzhou, P.R.China
This study was aimed to prepare mangiferin monosodium salt and compare its pharmacological effects with mangiferin. Mangiferin monosodium salt was prepared by are action of mangiferin and NaHCO3.Its structure was characterized by ESI-MS spectra and elemental analysis. Its pharmacological effects were compared with mangiferin. The results showed that mangiferin monosodium salt was first prepared, its pharmacological effects were better than mangiferin in anticough, antiexpectorant and antiinflammation effects. In conclusion, the synthetic route is practical with high yield mangiferin monosodium salt is expected to develops a potential drug for respiratory tract.
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Study On Mango Leaf and Mangiferin Protective effect of Mangiferin dropping pills on chronic liver injury in rats
Xiaoou Huang1, Jiagang Deng 2, Zhuang Chen1
1 2
Ruikang Hospital,Guangxi Traditional Chinese Medical University, Nanning, P.R.China Guangxi Traditional Chinese Medical University,Nanning, P.R.China
This experiment was aimed to study the protective effect and mechanism of Mangiferin dropping pills on chronic liver injury. The chronic liver injury models were induced by CCl4 in rats. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), totalprotein (TP), Albumin(ALB),
Hydroxyproline (HyP) in serum, malondiadehyde (MDA) content,superoxide dismutase (SOD) and glutathione peroxidase(GSH-PX) activities in liver homogenate were assayed by spectrophotometry;serum hyaluronic acid (HA) and procollagen PC were determined by radioimmunoassay; the transforming growth factor-l (TGF-l) Expression level were determined by radioitnmunoassay; hepatic tissue sections were stained with hematoxylin and eosin and examined under a light microscope. The result showed that: Mangiferin dropping pills was found to significantly decrease the serum transaminase activities and level of HA, Hyp and PC ( P<0.0l or P<0.05 ) , and improved the level of ALB,TP ( P<0.01or P<0.05) Meanwhile mangiferin dropping pills also decreased MDA content and prevented the reduction of SOD, GSH-PX activities in liver homogenate( P<0.01or P<0.05 ) TGF-1expression was inhibited after administration of mangiferin dropping pills. The results showed that mangiferin dropping pills could improve the pathological changes of liver tissue. The conclusions suggest that mangiferin dropping pills have protective effect on chronic liver injury in rats. The protective effect may be related to its antioxidant activity and the TGF-l expression level.
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Study On Mango Leaf and Mangiferin Effects of Mangiferin on cell cycle status and cyclin A,cyclin B1 expression of K562 cells
Zhigang Peng 1, Jun Luo 1,Yongrong Lai1,Shanjun Song2
1 2
The First Affiliated Hospital , Guangxi Medical University, Nanning , P.R.China Affiliated Union Hospital, Tongji Medical College, Huazhong University of Science and Technology,
Wuhan, P.R.China
This experiment was aimed to study the effect of mangiferin on cell cycle status and Cyclin A, Cyclin B1 expression in K562 cells , further to study the molecular mechanism for treating leukemia. The change of cell cyclestatus in K562 cells treated with mangiferin was performed by the flow cytometric; The expression of Cyclin AmRNA and Cyclin B1 mRNA was detected by semi-quantitative RT-PCR.The results showed that : K562 cells in G2/ M phase increased in a dose- dependent way after 24 hours mangiferin admininistration, indicating G2/ M phase blockage ;the expression of Cyclin A mRNA and CyclinB1mRNA enhanced in a dose-dependent way. The finding suggest that Mangiferin blocked the cell cycle progression in G2/ M phase ,and it can significantly increase the the expression of Cyclin A and Cyclin B1at the mRNA level in K562 cells , G2/ M phase blockage may be its one of the molecular mechanism for treating leukemia.
Effects of Mangiferin on myocardial ischemia induced by pituitrin in mice

Jianquan Wei, Zimin Zheng, Yong Pan, Ying Luo, Jian Huang, Shu Lai, Xiaolin Xu Departmentof Pharmacology, Youjiang Medical College for Nationalities, Baise, Guangxi, P.R.China
This study aimed to evaluate the protection of mangiferin against mice myocardial ischemia induced by pituitrin (PIT). Acute myocardial ischemic models were obtained by intraperitoneal administration of PIT 30u/kg in mice. Meanwhile, the mice were also intraperitoneal administration with different concentrations of mangiferin once daily for 7 days. The fluctuations of lead II on ECG, the myocardial SOD and MDA, the serum levels of LDH and CK were measured. The results showed that mangiferin could ameliorate t he abnormal changes of ECG (P<0.05),and up-regulate SOD activity, down-regulate myocardial MDA, serum CK and LDH levels(P<0.05 or 0.01). In conclusion, mangiferin can protect against myocardial ischemia induced by PIT in mice.
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Study On Mango Leaf and Mangiferin Effects of Mangiferin on Gastric Ulcers in Rats
Jianquan Wei, Zimin Zheng, Qiang XueYong Pan, Xiaolin Xu Pharmacy Teaching and Research office, Youjiang Medical College for Nationalities, Baise, Guangxi, P.R.China
This experiment was aimed to study the effect of mangiferin on gastric ulcers in rats. Ulcer index, area of ulcergastric juice volume, acidity and pepsin output were measured on experimental gastric ulcer induced by water-immersion stressacetic acid and pyloric ligation in rats. The changes of superoxidase dismutase (SOD) and malondialdehyde (MDA) in the gastric mucosa of gastric ulcer induced by waterimmersion stress were assessed. The results showed that:Mangiferin could inhibit the formation of ulcer, could increase the activity of SOD and decrease the contents of MDA in the gastric mucosa. acidity and pepsin output. The finding Mangiferin was not shown to be able to reduce gastric juice volume suggest that:Mangiferin has antiulcer effect in rats.
Effect of Mangiferin on the arachidonic acid metabolites in rat

Jiagang Deng, Li Yan, Licheng Guo Guangxi Traditional Chinese Medical University, Nanning, P.R.China
The study aimed to investigate the effect of mangiferin on arachidonic acid metabolites so as to elucidate its mechanisms of anti-inflammatory action. Rat air-pouch acute inflammatory model was established with intrathoracically injecting carrageenan. Leukot rieneB4 (LTB4), LTC4 and 6-ketoprostaglandin F1 alpha (6 - keto - PGFla) were measured using radioimmunoassay and reversed phase high performance liquid chromatography (RP - HPLC) method respectively. The results showed that mangiferin (10 and 100mol/L) was found to significantly decreased the contents of 6-keto-PGFla. However, the three concentrations of 1.0, 10 and100mol/L mangiferin did not affect the levels of LTB4 and LTC4. This finding suggest that the anti- inflammatory effect of mangiferin might be associated with inhibitory effect on 6- keto-PGFla .
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Study On Mango Leaf and Mangiferin Cardioprotective Effects of Mangiferin on Myocardial in Schemia Reperfusion Injury in Rats
Chao Wang, Guoxian Wang Department of pharmacology Jinzhou Medical College, Jinzhou, P.R.China
The study aimed to investigate the effect and possible mechanism of mangiferin on myocardial ischemia reperfusion (MIR) injury in rats. Forty-eight SD rats were randomly divided into six groups: sham group, model group , mangiferin groups of 10 mg/kg , 20 mg/kg , 40 mg/kg and Diaoxinxuekang group. Each group were injected with corresponding concentrations of mangiferin, Diaoxinxuekang or equal volume of saline gastrogavaged for 21d. One hour after the last administration myocardial is chemia reperfusion models were obtained by ligated left anterior descending coronary artery 40 minutes and followed by 120 minutes reperfusion and sham group was given all the procedures except ligation. Ultrastructure of myocardium with TEM and infiltration of polymorpho nuclear neutrophils (PMN) in myocardium with myeloperoxidase (MPO) were observed; serum activity of lactate dehyd-rogease (LDH), myocardial activity of superoxide dismutase (SOD) and contents of malondialdehyde (MDA) were detected respectively. The results showed that mangiferin could improve myocardial pathology and ultrastructurein mangiferin10 mg/kg group the activities of SOD was higher (P< 0.05) ; the contents of MPO, MDA , LDH were significantly lower ( P<0.05) . In conclusion, mangiferin can protect MIR. The myocardial protective mechanism of it may be realized by enhancing the activation of SOD activity, enhancing myocardial antioxygen capability, stabilizing myocardial cellular membrane and alleviating infiltration of polymerrphonuclear neutrophils (PMN) in myocardium.
Effect of Mangiferin on lymphocyte proliferation in immunosuppressed mice

Jiagang Deng, Ke Yang, Li Yan, Licheng Guo, Huiqin Tang Guangxi Traditional Chinese Medical University, Nanning, P.R.China
The immunodepressed mice were established by means of subcutaneous injection of cyclophosphamide (CTX). The mice of administration groups were given doses of 200 mg/kg, 100 mg/kg and 50 mg/kg mangiferin respectively and control group and model group were set up. MTT method was used to detect lymphocyte proliferation induced by ConA (5g/ml, 10g/ml). These findings suggest that mangiferin could significantly enhance ConA-stimulated T lymphocytes proliferation.
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The effects of Mangiferin on human platelet aggregation and secretion of CD62P

Yanping Huang Department of Hemotology, the First Affiliated Hospital, Guangxi Medical University, Nanning, P.R.China
The research investigated the effects of mangiferin on human platelet aggregation and expression of CD62P in secretion after platelet activation in vitro, and studied its mechanism of anti-aggregation. Whole blood samples was prepared from healthy adult volunteers, incubating with the drug at 37 , platelet activation was induced by collagen or arachidonic acid, the platelet aggregation was determined by implement's method; the percentage of CD62P-positive platelets was detected by flow cytometry. The result showed that 100g/ml mangiferin didn't influence the platelet aggregation when incubated with whole blood at 5min, 15min and 30min.Mangiferin could inhibit the platelet aggregation, the inhibitory effect was in a concentration-dependent manner. The amplitude of 4g/ml,20g/ml and 100g/ml mangiferin induced by collagen were 17.404.01 ohm, 15.604.06 ohm and 12.504.93 ohm, the idex induced by arachidonic acid were 28.207.32 ohm, 21.507.41 ohm and 16.607.03 ohm, the groups of 20g/ml and 100g/ml mangiferin showed significance with control group (P0.05).The inhibitory effects induced by collagen displayed weaker than that induced by arachidonic acid at the same concentration, the IC50were 140.77g/ml and 98.61g/ml, respectively. It could prolong the lagtime,100ug/ml mangiferin induced by arachidonic acid showed significance compared with sodium chloride(P0.05).But it did not influence the expression of CD62P induced by both agonists, the percentage of CD62P-positive platelets in collagen-induced platelet activation were 13.415.53%, 15.328.76%and 14.296.60%, which were 12.2911.48%, 12.129.52% and 13.9012.83% after stimulation with arachidonic acid, there was no difference between control and mangiferin-treated samples (P0.05). In conclusion, the present study demonstrates that mangiferin didnt influence the platelet aggregation when incubated with whole blood at different time, Mangiferin could inhibit platelet aggregation, the effect was more powerfully induced by arachidonic acid than that by collagen, but it did not affect the expression of CD62P in the course of platelet secretion.
126
Study On Mango Leaf and Mangiferin Effect of Mangifer on Serum E-cadherin, carcinoembryonic antigen and monoamine oxidase activity and cell cycle in live tumor rats
Huayi Huang 1, Caozan Nong 1, Lingxiao Guo 1, Shiyuan Zhao1, Xiliang Zha 2
1 2
Guangxi Minzu Hospital, Nanning, P.R.China Shanghai Medical College, Fudan University, Shanghai, P.R.China
Recent studies showed that mangiferin could improve immune function and had antioxidant and anti-tumor effects. In this study, we examined the anti-tumor effects of mangiferin and its mechanisms in rat. Wistar rats were implanted with Walker-256 tumor in the liver. Serum E-cadherin (sE-cad) and carcinoembryonic antigen (CEA) levels and monoamine oxidase (MAO) activity were measured by an enzyme immunoassay (ELISA). The sE-cad and MAO levels decreased and serum CEA levels were not change, when the dose of mangiferin was 15mg/kg per day. The results proved that mangiferin could affect the E-cadherin-mediated cell adhesion system and slow down the process of liver cirrhosis.
The Impact of Mangiferin on Releasing of Slow Reacting Substance of Anaphylaxis from Guinea-Pig Lung Tissue
Jiagang Deng*, Lichen Guo , Zuowen Zheng, Ke Yang, Li Yan Guangxi Traditional Chinese Medical University, Nanning, P.R.China
The aim of the present study is to investigate the mechanism of anti-inflammation action of mangiferin. Guinea pigs were intragastric administrated with different dosages of mangiferin during three weeks. Either the content of slow reacting substance of anaphylaxis (SRS-A) in lung tissue of guinea pig, and contractions of the isolated guinea pig ileum, were measured in the end of the third week. Results showed that releasing of SRS-A from guinea-pig lung tissue was depressed by mangiferin. Thus, we suggest that there is possibly a relationship between the depression of releasing SRS-A caused by mangiferin and anti-inflammation mechanism of mangiferin.
127
Study On Mango Leaf and Mangiferin Effects of on Mangiferin HBsAg and HBeAg Excreted by 2215 Cell
Zuowen Zheng*, Jiagang Deng, Ke Yang Guangxi Traditional Chinese Medical University, Nanning, P.R.China
Mangiferin has exhibited liver protection, antiasthmatic, antitussive and antivirus properties. The research observed the cytotoxicity of mangiferin and its effects on HBsAg and HBeAg secretion. The cytotoxicity tests were carried out on 2215 cell by MTT method. The inhibitory effect on HBsAg and HBeAg were detected in the nontoxic concentration. The TC50 concentration and the maximum nontoxic concentration of mangiferin were 250g/ml and 125g/ml respectively. Mangiferin had obvious inhibitory effect on HBeAg. The IC50 concentration of mangiferin was 37.6g/ml and the therapeutic index was 6.65.
The Effect of Mangiferin on hTERT-mRNA Expression and Telomerase Activity in K562 Cells
Zhigang Peng 1, Jun Luo 1, Yongrong Lai 1, Yuying Lu1, Shanjun Song 2
1
Department of Hemotology, The First Affiliated Hospital, Guangxi Medical University, Nanning, P.R.China
This study was designed to investigate the effects of mangiferin on hTERT- mRNA expression and telomerase activity in K562 cells and their relationship. The hTERT- mRNA expression was determined by RT- PCR assay in untreated and mangiferin- treated K562 cells, and telomerase activity was analyzed by TRAP- PCR- ELISA assay. The hTERT- mRNA expression of K562 cells was significantly inhibited when treatment with mangiferin, while telomerase activity was decreased. Mangiferin can inhibit the hTERTmRNA expression and telomerase activity of K562 cells. Telomerase activity might be related to the expression of hTERT- mRNA.
128
Study On Mango Leaf and Mangiferin Protective effect of Mangiferin on Alcohol-Induced Liver Injury in Mice
Jianquan Wei, Zimin Zheng, Yong Pan, Ying Luo Department of Pharmacology of Youjiang Medical College for Nationalities, Guangxi, P.R.China
The present study was to investigate the effect of mangiferin on alcohol-induced liver injury in mice. Mangiferin (15, 30, 60mg/kg) was given to mice. Liver injury was evaluated by measured the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), Enzymes of the glutathione S-transferase (GST) and Superoxide Dismutase (SOD) in the serum and content of malondialdehyde (MDA) and triglyceride (TG) in liver homogenates. As a result, mangiferin significantly protected against alcohol-induced liver injury as evidenced by decreasing elevated serum ALT, AST and GST and lowering level of MDA and TG. In addition, Mangiferin could elevate SOD level. The protective effect of mangiferin on alcohol-induced liver injury was mainly due to its ability to attenuate oxidative stress.
Pharmacodynamic Study of Total Glycosides Tablet of Mango Leaves

Naiping Wang, Jiagang Deng, Haibin Huang, Xuejian Li Guangxi Traditional Chinese Medical University, Nanning, P.R.China
Mango leaves are the dry leaves of Mangifera indica L., character and taste are acid, sweet, cool, calm, it has a function of smooth to the body. In folk, people use it to treat cough. Clinic testify that it has effectiveness on cough and expectoration which is caused by bronchitis, and also have effective on preventing asthma to some extend. Mango leaves also the main material on producing Mango cough drop. In this study, total glycosides were extracted from mango leaves and were made in tablet. In this experiment, pharmacodynamic effects of total glycosides tablet were evaluated by the variety of animal models. The cough models induced by ammonia liquor in mice were used to observe the antitussive effects. The methods of phenel red execretion in mice were used to investigate the expectorant effects. Bronchial asthma model induced by histamine-acetylcholine in guinea pigs was used to observe the antiasthmatic effects. The croton oil- induced ear swelling test was used to study the anti-inflammation effects. The results showed that total glycosides tablet has the anti-asthma, antitussive and expectorant effect in vivo, which supplied for further research on pharmacological mechanisms.
129
Study On Mango Leaf and Mangiferin The influence of mangiferin on the body temperature of rabbit in endotoxin-induced fever
Jiagang Deng, Zuowen Zheng, Ke Yang Guangxi Traditional Chinese Medical University, Nanning, P.R.China
The research observed the influence of mangiferin on the body temperature of rabbit in endotoxin-induced fever. The healthy qualified Japanese white rabbits were chosen, endotoxin was injected intravenously to lead to fever model, mangiferin was administrated intragastricly and then the change of rabbits body temperature was supervised in 5 hours. The result showed that mangiferin has function of antipyretic on the animal fever model caused by endotoxin. High dosage of mangiferin is equal to complex aspirin.
The study on mangiferin protective role of lipid peroxidation damage of brain tissues in rats
Huayi Huang1, Ming Zhong2, Gang Meng1, Chaozan Nong1, Shiyuan Zhao1, Shengming Yu2, Linyun Huang2
1
Central Laboratory of Clinical Medicine Institute of Guangxi Hospital for Nationalities, Nanning,
P.R.China
2
Medicine Institute of Guangxi for Nationalities, Nanning, P.R.China
This study aimed to investigate the protection of mangiferin protective role of lipid peroxidation damage of brain tissues in rats. The brain damage model of rats was established by alloxan. Maleic dialdehyde (MDA) levels, Lipid Peroxide (LPO) and Superoxide Dismutase (SOD) activities were determined to assess lipid peroxidation. The result showed that the activity of SOD in brain tissue decreased both in blank group and model group. Compared with model group, mangiferin group had significantly difference(P<0.05 ).The content of LPO in mangiferin high dose group was decreased. Compared with model group, it had significantly difference (P<0.05 ). The activity of MDA of mangiferin group were lower than model group, but there was no difference (P>0.05). In conclusions, mangiferin has protective effect on lipid peroxidation damage in rats.
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Study On Mango Leaf and Mangiferin The antidepressant effect of mangiferin on the behavioral despair mice
Zimin Zheng, Jianquan Wei, Ying Luo, Yong Pan, Qiang Xue. Youjiang Medical College for Nationalities, Baise, Guangxi, P.R.China
The research investigated the anti-depressant effect and the possible mechanism of mangiferin in animal models. The forced swim test and the tail suspension test were applied to study the anti-depressant effect of mangiferin in mice. Mangiferin at the dose of 15, 30 and 60mg/kg significantly reduced immobility time on the tail suspension test, and at the dose of 60mg/kg significantly shorter immobility time in force swimming test. The present results suggested that mangiferin has significant anti-depressant effect.
Effects of Mangiferin on lipid peroxidation metabolism of blood in rats

Huayi Huang1,Ming Zhong2,Gang Meng1,Chaozan Nong1,Shiyuan Zhao1,Hongmei Zu2,Qingqing Zhan2, Zhijin Zhao1
1
Central Laboratory of Clinical Medicine Institute of Guangxi Hospital for Nationalities, Nanning, P.R.China
The Pharmacological Room of Medicine Institute of Guangxi for Nationalities, Nanning, P.R. China
This study aimed to investigate the effects of mangiferin on lipid peroxidation metabolism of blood in rats. The lipid peroxidation injury model of rats was established. The glutathione peroxidase (GSH-Px), and catalase (CAT) activities, the content of Lipid Peroxide (LPO) and hemoglobin (Hb) were determined to assess lipid peroxidation metabolism abilities. The result showed that: The activity of GSH-Px and CAT increased both in blank group and model group. Compared with model group, mangiferin group had significantly difference (P<0.01).The content of LPO in mangiferin high dose group was decreased. Compared with model group, it had significantly difference (P<0.01). Compared with blank group, the content of Hb in mangiferin group were lower than blank group, but there was no difference (P>0.05). In conclusion, mangiferin can decrease the level of lipid peroxidation in rat blood, its metabolism may relate to enhance the activities of antioxidant enzyme system and decrease the content of Hb.
131
Study On Mango Leaf and Mangiferin Antlviral activity of mangiferin against herpes simplex virus type 2 in vitro
Xuemei Zhu, Jiaxing Song, Zongzhi Huang, Yimou Wu, Mingjun Yu Department of Microbiology,Hengyang Medical College,Hengyang , PR China
The effect of mangiferina tetrahydroxy pyrrolidone saponin extracted from the leaves of mango(Mangifera indica) against herpes simplex virus type2 (HSV-2)in vitro was studied.The 50% effective concentration (EC50)of it against HSV-2 plaque for-mation in HeLa cells was 111.7gml-1,and the con-centrations of 33 and 80gml-1 reduced the virus replicative yields by 90 (EC90)and 99%(EC99),re-spectively.The therapeutic index (IC50 EC50) was 8.1. Mangiferin did not directly inactivate HSV-2.The results of the drug addition and removal tests suggest that mangifetin inhibits the late event in HSV-2 replication.
Effect of Mangiferin on the Arachidonic Acid Metabolizing Enzymes in Rat Neutrophils

Jiagang Deng, Li Yan Guangxi Traditional Chinese Medical University, Nanning, P.R.China
Mangiferin, a natural polyphenol is known to exhibit anti-inflammatory, antioxidant, and antiviral effects. Arachidonic acid (AA) metabolizing enzymes play an important role in case of inflammatory responses, including respiratory infections. In this study, we investigated the effect of mangiferin on the various family members of AA metabolizing enzymes. We measured activity of phospholipase A2 (PLA2) in rat air-pouch acute inflammatory model using acid-base titration and evaluated the activity of 5-lipoxygenase (5-LOX) and cyclooxygenases (COX) in pleural effusion using radioimmunoassay and reversed phase high performance liquid chromatography (RP-HPLC) method respectively. From this analysis, we found mangiferin significantly decreased the activities of PLA2 at 140, 70 and 35mg/kg, and significantly inhibited the activities of COX at 10mol/L and 100mol/L. However, the three concentrations of 1.0, 10 and 100mol/L, mangiferin did not affect the activities of 5-LOX. These results suggest that the anti-inflammatory effect of Mangiferin might be associated with its inhibitory effect on PLA2 and COX activity.
132
Study On Mango Leaf and Mangiferin Antiviral effect of virus

Minshi Zheng 1, Zongyi Lu2 Department of Microbiology, Jiangxi Medical, Nanchang, PR China Department of Traditional Medicine,Shanghai Researh Institute of Medicinal Industry, Shanghai, P.R. China
mangiferin and Isomangiferin on herpes simplex
Using tissue culture techniques the present study assured us of mangiferin
and isomangiferin in
the antiviral action. against HSV-l. Utilizing 4 main patterns fer evaluating drug effectivenss (ie intratube drug-on-virus directaction, simultaneous addition addition of drug-virus- inoculum to cell bottle, virus inoculation preceding drug addition,and drug addition followed by virus inoculation),it was readily found by logarithm determination of HSV-I inhibition that isomangiferin was superior to such control drugs as acyclovir, idoxuridine, and cyclocytidine in logarithm by 0.27-0.50, and that mangiferin was lower than isomangiferin in logarithm by 0.53. The average plaque reduction rates of mangiferin and isomangiferin were 69.5 and 56.8%, respectively. All in all the antiviral effect of mangiferin and isomangiferin was attributed presumably to their capability to inhibit virus replication within cells.
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Chemical Study and Analytical methods

Characterization and quantitative determination of the impurity in prepared mangiferin extracted from Mangifera indica L. leaves
Jieping Qin1, , Jiagang Deng1, Yuqi Feng2, Xu Feng1
1 2
Guangxi Traditional Chinese Medical University, Nanning, P.R. China Department of Chemistry, Wuhan University, Wuhan, P.R. China
Abstract Mangiferin is a pharmacologically active and a natural xanthone C-glycoside, which widely distributed in the bark, fruits, roots and leaves of Mangifera indica L. and many higher plants. Mangiferin has been reported to possess important pharmacological activity, including antioxidant, antitumor, antiviral, immunomodulatory, and antidiabetic activities. As the active pharmaceutical ingredients, mangiferin product extracted from M. indica L. leaves can be used to prepare medicines, such as antivirus and anti-inflammation medicament. In order to ensure the quality and safety of pharmaceuticals, it becomes important to determine the contents of mangiferin and related substance in the mangiferin products. In this paper, an unknown impurity in the prepared mangiferin extracted from Mangifera indica L. leaves was isolated and determined by a newly developed reverse phase high performance liquid chromatographic (HPLC) method. The unknown impurity was identified by LC/MS/MS and its structure was confirmed by spectrometric studies of IR and NMR. Based on the spectral data, the molecular structure of the unknown impurity was characterized as 2-C--D- glucopyranosyl-1, 6, 7trihydroxy-3-methoxy-xanthen-9-one. The fragmentation mechanisms of the impurity and mangiferin were proposed. From the comparison of the HPLC chromatogram between the prepared mangiferin product and the crude methanol extract of Mangifera indica L. leaves, it can be seen that the unknown impurity is natural occurring compound. The validated HPLC method was used to determine the amounts of the impurity in nine batches of prepared mangiferin products. The results shown the contents of the impurity in nine batches of mangiferin sample (mangiferin content >90%) were range from 13.3mgg-1~75.9 mgg-1.
Keywords: Mangifera indica L.; Prepared Mangiferin; Impurity; HPLC; LC/MS/MS; NMR; Characterization; quantitative determination.
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1 Introduction Mangiferin (Fig.1), molecular formula: C19H18O11, is a pharmacologically active and a natural xanthone C-glycoside, which widely distributed in the bark, fruits, roots and leaves of Mangifera indica L. and in more than a hundred species of higher plants [14]. Mangiferin has been reported to possess important and antiviral[10,12], antioxidant[1318],
24]
broad pharmacological activity, including antidiabetes[57], antitumor [811], immunomodulatory[3,1922], anti-inflammatory

[22]
and vascular modulatory activity[23,
. As the active
pharmaceutical ingredients, mangiferin product extracted from M. indica L. leaves can be used to prepare medicines, such as antivirus and anti-inflammation medicament. In order to ensure the quality and safety of pharmaceuticals, it becomes important to determine the contents of mangiferin and related substance in the prepared mangiferin products from M. indica L. leaves. Although various analytical methods have been developed for the separation and quantitative determination of neomangiferin and mangiferin in Rhizoma Anemarrhenae[25, 26]. There has been no report in the literature on the identification and characterization of the unknown compound in mangiferin extracted from M. indica L. leaves even though it has been found in capillary zone electrophoresis(CZE) analysis before
[27, 28]
. This paper aims at the isolation and
characterization of the impurity in mangiferin product extracted from M. indica L. leaves. A new reversed phase HPLC method was developed for the isolation and quantitative determination of mangiferin and the impurity. The HPLC chromatogram of prepared mangiferin contained two peaks, one major peak and another minor peak follows. According to the control mangiferin 1H NMR and HPLC-DAD results, the front major peak represents mangiferin, while the rear minor peak was an unknown impurity. A comprehensive study was undertaken for the identification of the impurity using LC/MS/MS followed by further characterization by various spectroscopic techniques. Based on the spectral data, the molecular structure of the impurity was characterized as 2-C--D-glucopyranosyl-1, 6, 7 -trihydroxy-3-methoxyxanthen-9-one. The validated HPLC method was used to determine the amounts of the impurity in nine batches of prepared mangiferin products extracted from the M. indica L. leaves. The results shown the contents of the impurity in nine batches of mangiferin sample were range from 13.3mgg-1~75.9 mgg-1.
R HO 4` HO HO 6` O 5` 2` 1` OH 3` OH O 2 1 8 3 4 4a 8b O 4b 8a 7 OH 5 6 OH
Fig.1 Chemical structure of mangiferin and impurity. 1: R=OH (mangiferin); 2: R=OCH3 (impurity).
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2 Experimental methods 2.1. Liquid Chromatographic Systems Analytical An Agilents 1100 Series HPLC Separation module equipped with a diode array UV detector was used. A Hypersil C18 column (150mm4.6 mm, i.d., 5m particles) (Elite Co. Ltd., China) was used for the separation. The mangiferin samples and each preparative peak fraction were analyzed by HPLC. The mobile phase consisting of A: 0.1%phosphoric acid (v/v in H2O) and B: HPLC grade acetonitrile, with a timed gradient program T (min) / % B: 0/10, 15/12, 20/15, 30/45, and a flow rate of 1.0 mL per min was used. The injection volume was 5L and the detector wave-length was fixed at 258 nm. The column was maintained at 30C throughout the analysis. Preparative A Shimadzu preparative HPLC equipped with LC-8A pump, SCL-8A System controller, SPD-6AV UVVIS detector, Rheodyne Injector with 1 mL sample loop was used. The data were collected and processed using Shimadzu CR7A chromatopak. A Lichrospher-C18 column( 250mm10.0 mm, 10m) (Jiangsu Hanbang Science & Technology Co., Ltd., China) was employed for the separation. The mobile phase was consisted of Solvent A (contained 0.05% trifluoroacetic acid in water) and solvent B (HPLC grade methanol) in a ratio of 65:35 (v/v). The flow rate was set at 2.0 mL/ min. Detection was carried out at 258 nm. Column temperature was ambient. 2.2. mass spectrometry(MS/MS) A Bruker LC-micro TOF-Q electrospray ionization mass Spectrometer was used to analyze mangiferin and the unknown compound, respectively. The MS was operated in the negative ionization mode from the electrospray ionization (ESI) source. The temperature of the drying gas (N2) was 200 C and flowed at 3.0 L/ min. The nebulizing pressure (N2) was maintained at 1.0 bar to generate the spray. The MS/MS data was generated with the collision energy ramping from -10 to -60V in nitrogen atmosphere. The mass to charge ratio was scanned across the range of m/z 50500. VEnd plate offset = 500V. The mass spectrometer was operated in a manner similar to the LCMS experiments, except that a target mass (parent ion) was mass selected and separated from all other ions. This mass selected parent ion was then fragmented in collisions with nitrogen gas in the trap. Then, the fragment ions were analyzed in the negative mode.
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2.3. NMR spectroscopy NMR measurements were performed on a VARIAN INOVA-600MHz instrument (both for 1H and 13C) at 25C in DMSO-d6. The 1H and 13C chemical shift values were reported on the scale in ppm, relative to TMS (= 0.00) as internal standard. Assignments were further confirmed by running two-dimensional chemical shift correlation experiments. 2.4. FT-IR spectroscopy The IR spectra of isolated impurity and mangiferin were recorded by a NICOIET NIXUS-470 FT-IR spectrophotometer using KBr method at 25 2.5. Rotary Evaporator A RE-52CS Rotary Evaporator (Shanghai Yarong Biochemistry Instrument Factory, Shanghai, China) was utilized to evaporate solvent. 2.6. Chemicals Dimethyl sulphoxide-d6 (for NMR) was purchased from Aldrich Chemical Co., USA. HPLC grade acetonitrile was purchased from Tianjin Shield Company of China. All other chemicals and reagents used were of analytical grade unless indicated otherwise. 2.7. Mangiferins Control mangiferin purchased from Sigma (SigmaAldrich, St. Louis, MO 63103 USA 314-771-5765), Cat. No.M3547, with purity > 99.9% by HPLC method. Mangiferin drug samples extracted from Mangifera indica L. leaves was supplied by Pharmaceutical Factory of Guangxi Traditional Chinese Medical University (China). 2.8. Sample Preparation The mangiferin samples injected for LC analysis were all dissolved in the solvent consisting of methanol-water (1:1, v/v). All sample solutions were filtered through a 0.45m filter before injection. The concentrations of mangiferin sample were approximately 1 mg/mL for preparative LC and the injection volume was 1ml. For analytical LC, the concentrations of mangiferin samples were about 0.6 mg/mL. The injection volume was 5l.
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3 Result and discussion 3.1. Analysis of mangiferin sample by HPLC-DAD An Agilents 1100 Series HPLC Separation module equipped with a diode array UV detector described in Section 2.1 was used. The HPLC chromatogram of prepared mangiferin product contained two peaks, one major peak and another minor peak follows. According to the control mangiferin 1H NMR, LCMS and HPLC-DAD results, the front major peak represents mangiferin. HPLC analysis of nine batches of mangiferin samples revealed the presence of an impurity at RRT 1.34 with respect to principle peak. The typical HPLC chromatogram of prepared mangiferin product and UV spectra of mangiferin and the impurity are shown in Fig. 2. It can be seen that the UV spectra of mangiferin (max = 240,258,317 and 367nm ) and the impurity (max = 238,258,319 and 367nm ) are very similar and display the characteristic of xanthone, which suggest the two structures were quite similar.
Time (min) Fig. 2. HPLC chromatogram of prepared mangiferin product and their UV spectra ( ) mangiferin ; ( ) impurity. 3.2. Isolation Procedure A Shimadzu preparative HPLC System was used. The unknown impurity in prepared mangiferin product was separated and purified under the optimum reversed-phase preparative HPLC condition described in Section 2.1. In this separation process, the sample solution was injected into the separation column. The separation temperature was ambient. The effluent from the outlet of the column was continuously monitored at 258 nm. Each peak fraction was manually collected according to the chromatogram. The typical preparative HPLC chromatogram is shown in Fig. 3.
138
Absorbance (mAU)
Time (min)
Fig. 3. Preparative HPLC chromatogram of mangiferin product In the first separation, the peak fraction of the impurity was found contain mangeferin by HPLC analysis. So the second separation must been performed. The second preparative HPLC chromatogram was shown in Fig. 4. In the second separation, the peak fraction of the impurity was manually collected according to the chromatogram and evaporated under reduced pressure. In the end, some pale yellow needle crystals were obtained and then dissolved in 1:1 methanol-water for HPLC analysis to confirm to be a pure compound.
Absorbance (mAU)
Time (min)
Fig. 4. Preparative HPLC chromatogram of the unknown compound purification 3.3. Identification of the unknown compound by/MS/MS A Bruker LC-micro TOF-Q mass Spectrometer was used to analyze mangiferin and the unknown compound, respectively. The experimental method was described in Section 2.2. The mass spectra obtained in the negative mode. The mass spectrum of mangiferin (Fig. 5) shows a [M-H] molecular ion peak at m/z 421.0775 (C19H17O11,calculated:421.0776), while the mass spectrum of the unknown compound (Fig. 6)
139

exhibits a [M-H] molecular ion peak at m/z 435.0973 (C20H19O11,calculated:435.0933), which is 14 mass unitsCH2 more than that of mangiferin. The increase of 14 mass units may be caused by methylation .
Fig. 5.
HR-ESI mass spectrum of mangiferin.
Fig. 6.
HR-ESI mass spectrum of the unknown impurity
140

To obtain additional structural information, MS/MS analysis was further performed. The
HR-ESI-MS/MS spectrum of the unknown impurity (Fig.7 (A) exhibited a series of product ions:m/z 345.0657 C17H13O8,calculated:345.0615 ,315.0486 C16H11O7,calculated:315.0510 ,287.0189
C14H7O7,calculated:287.0197 ,272.0308 C14H8O6,calculated:272.0326 , while the spectrum of

mangiferin exhibited a series of product ions: m/z 331.0436C16H11O8,calculated:331.0459,301.0335
C15H9O7,calculated:301.0354,271.0237C14H7O6,calculated:271.0248 (Fig. 7 (B) ). These indicated

the two compounds were very similar structurally. Fragment ions of the unknown compound at m/z 435,345 and 315 were assumed to be [421+ CH2 ] , [331+CH 2] and [301+CH2 ] respectively, which suggest the hydroxylation of mangiferin at positions 3 could be processed through O-methylation (m/z 14) at hydroxylated group to produce methylmangiferin with m/z of 435 (421+14). The formation of these product ions can be explained by the dissociation mechanism shown in Fig. 8 (mangiferin) and Fig. 9 (impurity).
A ) impurity
B ) mangiferin
Fig. 7.
HR-ESI MS/MS spectra of the unknown impurity (A) and mangiferin (B).
141

HOH2 C O OH OH OH OH OH O HO O
OH
mangiferin
M.Wt .422
-H
HOH2C O OH
HO
OH
HO
OH
- C6H10O5
O
OH OH
OH
m/z 421
O OH O
- C3H6O3
m/z 259
HO
OH HOH2C O
m/z 331
OH
OH
- C H 2O
O O OH O m /z 3 01
O O OH O
m/z 331
O CH2OH
- C H 2O
- CH2O
HO
OH
HO
OH
OH
O
- CH2O
O
m/z 301
O O
m/z 271
OH
m/z 271
Fig. 8. Fragmentation mechanism for the product ions from M-H peak of mangiferin.
142

HOH2C O OH OH OH OH OH O H3CO O OH
impurity
M.Wt .436
-H
HOH2C O OH
H3CO
OH
O O
O
OH
OH OH OH O
m/z 435
HO
O OH O
-C3H6O3
H H2 CO O HOH2C O OH O
m/z 287
OH
-C2H4
OH
m/z 345
O OH OH O
m/z 315
OH
-CH2O
HOH2C OH OH O
m/z 345
H3CO
OH
GLUCOSE OH OH O
H
H3CO O
impurity M.wt.436
OH
GLUCOSE OH O O
m/z 435
-GLUCOSE
O
H3CO
OH
OH
-O
O
m/z 272
Fig.9
Fragmentation mechanism for the product ions from M-H peak of the impurity
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3.4. Infrared spectrum analysis Fig. 10 shows the infrared spectrum of the unknown impurity. The main absorbance peaks included 3521,3420, 1647, 1613, 1480, 1405, 1289, 1210, 1210, 1176, 1089, 808 cm
In the IR
spectrum,,absorption characteristics of a xanthone were observed at 1647 cm 1 (conjugated carbonyl), as well as 1613, 1480 cm 1 (aromatic ring C=C stretching). The peaks at 3521, 3420 cm 1 corresponding to
OH stretching suggested the presence of hydroxide bond in molecular structure.
Fig. 10.
Infrared spectrum of the unknown compound.
3.5. NMR analysis Assignments of the 1H NMR and 1C NMR resonances of mangiferin and the unknown impurity were obtained by using Varian spectra recorder described in Section 2.3. 1HNMR analysis of the unknown impurity suggested the presence of three chelated hydroxyl group ( 13.7,10.7 and 9.83 ppm), which were assigned to the protons of C1-OH,C6-OH and C7-OH. In the 1H NMR spectrum of the unknown impurity, it can be seen three single aromatic protons (6.64, 6.90 and 7.41 ppm),these aromatic protons were assigned to the protons at C-4, 5 and 8 positions of the xanthone [26]. In contrast with the 1H NMR spectrum of mangiferin, the obvious difference was the presence of a methyl singlet at 3.87 and 3.90 ppm. In the 13C NMR spectrum, appearance of downfield methoxyl signals at 56.8 and 57.1 ppm indicated the presence of a methoxy group. The paired resonances during the NMR examination could be explained by the influence of a conformational isomer caused by spin hindrance of ring structure, cause their magnetic inequivalence.
144

Based on the spectroscopic analysis above,the structure of the unknown impurity was characterized as 2-C--D-glucopyranosyl-1, 6, 7 -trihydroxy-3-methoxy-xanthen-9-one. The
1
H-NMR and
13
C-NMR
spectrum of mangiferin and the unknown impurity are presented in Fig. 11 and Fig.12. The HSQC spectrum of the unknown impurity is presented in Fig.14. The 1H-NMR and shown in Table 1.
13
C NMR spectrum data are
A) mangiferin
13
12
11
10
ppm
B) impurity
13
12
11
10
ppm
Fig.11
HNMR spectrum of the mangiferin (A) and impurity (B)
145
A) mangiferin
B) unknown compound
180
170
160
150
140
130
1 20
110
100
90
80
70
60
50
40
ppm
Fig. 12.
13
CNMR spectra of the mangiferin (A) and unknown compound (B).
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Table 1
1
HNMR and 13CNMR data for mangiferin and the unknown compound position mangiferin unknown compound H 1 2 3 4 4a 4b 5 6 7 8 8a 8b 9 1` 2` 3` C 162.5 108.3 164.5 94.0 156.9 151.4 103.3 154.7 144.4 108.7 112.4 102.0 179.8 73.7 82.3 71.3 70.9 79.7 62.2 H C 162.0(161.3) 108.3(108.5) 164.9(166.1) 90.4 (91.2) 157.4(157.5) 151.5(151.6) 103.0(103.2) 154.9 144.5 108.7(109.1) 112.5 102.5(102.7) 179.7(180.1) 73.5(73.3) 82.3(82.4) 71.0(70.3) 71.5(71.6) 79.7(79.8) 62.4
6.34(1H,s)
6.64(1H,s)
6.83(1H,s)
6.90(1H,s)
7.35(1H,s)
7.41(1H,s)
4.60(1H,d,J=9.6)
4.58(4.64) 3.15 3.99(4.21) 3.09 3.20 3.70,3.37 13.7(1H,s)
4` 3.104.10(5H,m) 5` 6` 1-OH 13.7(1H,s) 3-OH 10.6(1H,s) 6-OH 10.7(1H,s) 7-OH 9.78(1H,s) 3-OMe
10.7(1H,s) 9.83(1H,s) 3.90(3.87)(3H,s)
57.1(56.8)
3.6. HPLC analysis of the impurity in mangiferin extracted from Mangifera indica L. leaves. In this paper, nine batches of mangiferin products (mangiferin content >90%) were determined by using the HPLC analytical method described in Section 2.1. The method validation test shown the calibration curve of impuryty was linear in the range of 0.96~19.2g/mL, A = 47742.82C1.66 (r=0.9999, n=5). The average recovery rate was 97.8%, and RSD = 2.2% (n=6). The impurity contents (mgg-1) of nine batches of mangiferin product extracted from M. indica L. leaves are listed in Table 2. From the determination results, it can be seen the impurity content variations lain in the different batches of mangiferin sample. This is probably due to the different collected month of crude drugs or the manufacturing processes.
147

Table2 Homomangiferin content (mgg-1) in 9 batches of mangiferin (n=3) impurity content (mgg-1) NO. 1 20050624 20050709 20050715 20060421 20060426 20060517 20060608 20060918 20070406 39.4 29.4 76.2 39.1 24.1 72.3 38.8 66.0 13.1 2 38.9 29.7 75.4 38.7 24.6 72.4 39.2 65.4 13.2 3 39.4 30.2 76.0 38.9 24.0 72.1 38.7 66.7 13.5 39.2 29.8 75.9 38.9 24.2 72.3 38.9 66.0 13.3 0.8 1.4 0.6 0.6 1.4 0.3 0.7 1.0 1.6
average (mgg-1)
RSD(%)
In order to investigate if the unknown impurity was the artifact during the preparing progress of mangiferin products, the HPLC analytical method described in Section 2.1 was used for the analysis of prepared mangiferin product and the crude methanol extract of Mangifera indica L. leaves. Their HPLC chromatograms were shown in Figure 13. It can be seen that this compound exist both in the prepared mangiferin and crude methanol extract of Mangifera indica L. leaves. Thus the unknown impurity is natural occurring compound.
Fig. 13.
HPLC chromatograms of prepared mangiferin (A) and of crude methanol extract of Mangifera indica L. leaves (B). 1. mangiferin ; 2. impurity
148

4 Conclusions An unknown impurity in mangiferin product extracted from Mangifera indica L. leaves was isolated and purified by preparative HPLC. Its molecular structure was identified by UV,FT-IR,HR-ESI/MS,MS/MS,
1
H NMR and
13
C NMR analysis. The spectroscopic analysis of the isolated unknown impurity indicated
that the molecular structure of the unknown impurity is hydroxylation of mangiferin at positions 3 through O-methylation at hydroxylated group to produce methylmangiferin derivative, as evident from MS/MS, FT-IR and NMR techniques. Based on the spectroscopic data,the structure of the impurity was characterized as 2-C--D-glucopyranosyl- 1,6,7- trihydroxy-3-methoxy-Xanthen-9-one. HPLC analysis of the impurity in 9 batches of mangiferin product extracted from M. indica L. leaves (mangiferin content >90% ) shown the contents of impurity in the different batches of mangiferin product were range from 13.3mgg-1~75.9 mgg-1. From the comparison of the HPLC chromatogram between the prepared mangiferin product and the crude methanol extract of Mangifera indica L. leaves, It can be seen the unknown impurity is natural occurring compound. 5 Acknowledgements This study was supported in part by Guangxi Science and Technology Department (No. KUI CAI JIAO 2007-109 and No. KUI CAI JIAO 2009-7). The authors wish to thank Dr. F. Li and Dr. X.J. Wu at Wuhan University for their kind help in the analysis of 1HNMR and 1CNMR spectra. We would also like to thank Y.Y Zhang, W.Zhang and W. Liu for their kind help in the separations. 6 References 1.R.A. Finnegan, R.A. Stephani, G. Ganguli, S.N. Ganguly, A.K. Bhattacharya,J. Pharm. Sci. 57 (1968) 1039. 2. E.H. Karunanayake, S.R. Sirimanne, J. Ethnopharmacol. 13 (1985) 227. 3. N. Makare, S. Bodhankar, V. Rangari, J. Ethnopharmacol. 78 (2001)133. 4.A.J. Nunez Selles, H.T. Velez Castro, J. Aguero-Aguero, J. Gonzalez-Gonzalez, F. Naddeo, F. De Simone, L. Rastrelli, J. Agric. Food Chem.50 (2002) 762. 5. Miura T, Ichiki H, Iwamoto N, Kato M, Kubo M, Sasaki H, Okada M, Ishida T, Seino Y, Tanigawa K (2001) Biol Pharm Bull 24(9):1009-1011. 6.D. Garcia, J. Leiro, R. Delgado, M.L. Sanmartin, F.M. Ubeira, Phytother.Res. 17 (2003) 1182. 7.S. Muruganandan, K. Srinivasan, S. Gupta, P.K. Gupta, J. Lal. Journal of Ethno- pharmacology 97 (2005) 497-501.
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8.N. Yoshimi, K. Matsunaga, M. Katayama, Y. Yamada, T. Kuno, Z. Qiao,A. Hara, J. Yamahara, H. Mori, Cancer Lett. 163 (2001) 163. 9.J.M. Leiro, E. Alvarez, J.A. Arranz, I.G. Siso, F. Orallo, Biochem. Pharmacol.65 (2003) 1361. 10. S. Guha, S. Ghosal, U. Chattopadhyay, Chemotherapy 42 (1996) 443. 11. A. Sarkar, Y. Sreenivasan, G.T. Ramesh, S.K. Manna, J. Biol. Chem.279 (2004) 33768. 12. M.S. Zheng, Z.Y. Lu, Chin. Med. J. (Engl.) 103 (1990) 160. 13.M. Yoshikawa, H. Shimoda, N. Nishida, M. Takada, H. Matsuda, J. Nutr. 132 (2002) 1819. 14.G.M. Sanchez, L. Re, A. Giuliani, A.J. Nunez-Selles, G.P. Davison, O.S. Leon-Fernandez, Pharmacol. Res. 42 (2000) 565. 15.S. Muruganandan, S. Gupta, M. Kataria, J. Lal, P.K. Gupta, Toxicology 176 (2002) 165. 16.G. Garrido, R. Delgado, Y. Lemus, J. Rodriguez, D. Garcia, A.J. Nunez-Selles, Pharmacol. Res. 50 (2004) 165. 17.G. Martinez, A. Giuliani, O.S. Leon, G. Perez, A.J. Nunez-Selles, Phytother. Res. 15 (2001) 581. 18.T. Miura, H. Ichiki, I. Hashimoto, N. Iwamoto, M. Kato, M. Kubo, E.Ishihara, Y. Komatsu, M. Okada, T. Ishida, K. Tanigawa, Phytomedicine 8 (2001) 85. 19. J. Leiro, J.A. Arranz, M. Yanez, F.M. Ubeira, M.L. Sanmartin, F. Orallo, Int. Immunopharmacol. 4 (2004) 763. 20. U. Chattopadhyay, S. Das, S. Guha, S. Ghosal, Cancer Lett. 37 (1987) 293. 21.D. Garcia, R. Delgado, F.M. Ubeira, J. Leiro, Int. Immunopharmacol. 2 (2002) 797. 22.J. Leiro, D. Garcia, J.A. Arranz, R. Delgado, M.L. Sanmartin, F. Orallo, Int. Immunopharmacol. 4 (2004) 991. 23.A.E. Beltran, Y. Alvarez, F.E. Xavier, R. Hernanz, J. Rodriguez, A.J.Nunez, M.J. Alonso, M. Salaices, Eur. J. Pharmacol. 499 (2004) 297 24.S. Prabhu, Mallika Jainu, K.E. Sabitha, C.S. Shyamala Devi. Vascular Pharmacology 44 (2006) 519525 25.J. J. Wang, Z. Y. Lou, Z. Y. Zhu, Y.F. Chai, Y.T. Wu, Chromatographia, 2005, 61, June (No. 11/12)633-636 26.Qinghua Sun, Ailing Sun, Renmin Liu. J. Chromatogr. A 1104 (2006) 6974. 27. H. Huang, C. Nong, L. Guo, S. Zhao, X. Zha, Zhongguo Yiyuan Yaoxue Zazhi. 22 (2002) 3. 28.Chaozan Nong, Weisheng He, Debbi Fleming, Lili Pan, Huayi Huang, Journal of Chromatography B, 826 (2005) 226-231.
150
Study On Mango Leaf and Mangiferin A new C-glycosyl xanthone isolated from Davallia solida
Sandrine Rancon1, Annie Chaboud1,*, Nicole Darbour1, Gilles Comte2, Denis Barron2, Jean Raynaud1, Pierre Cabalion3 1 Laboratory of Pharmacognosy, Faculty of Pharmacy, Lyon, France 2 Natural Products Laboratory, Claude Bernard University of Lyon, Villeurbanne, France 3 ORSTOM, UR 4G, Departement Sante, Paris, France
A new xanthone glycoside has been isolated together with mangiferin from Davallia solida. The structures were elucidated by chemical and spectral means as 2-C--D-glucopyranosyl-1,3,6,7 -tetrahydroxyxanthone (mangiferin) and 2-C--D-xylopyranosyl-1,3,6,7-tetrahydroxyxanthone.
Characterization and quantitation of polyphenolic compounds in bark, kernel, leaves, and peel of mango (Mangifera indica L.)
Jacqueline C. Barreto,Maria T.S. Trevisan,William E. Hull, Gerhard Erben,Edy S. DE Brito, Beate Pfundstein, Gerd Wu Rtele, Bertold Spiegelhalder, and Robert W. Owen German Cancer Research Center, Heidelberg, Germany
The contents of secondary plant substances in solvent extracts of various byproducts (barks, kernels, peels, and old and young leaves) in a range of Brazilian mango cultivars were identified and quantitated. The results show that the profiles of secondary plant substances such as xanthone C-glycosides, gallotannins, and benzophenones in different byproducts vary greatly but are fairly consistent across cultivars. The free radical scavenging activity of the solvent extracts was evaluated using a high-performance liquid chromatography-based hypoxanthine/xanthine oxidase assay and revealed dose-dependent antioxidant capacity in all extracts. Four (mangiferin, penta-O-galloylglucoside gallic acid, and methyl gallate) of the major phenolic compounds detected were also evaluated in additional in vitro bioassay systems such as oxygen radical absorbance capacity, 2,2-diphenyl-1-picrylhydrazyl, and ferric reducing ability of plasma. Mangiferin in particular, detected at high concentrations in young leaves (Coite ) 172 g/kg), in bark (Momika ) 107g/kg), and in old leaves (Itamaraka ) 94 g/kg), shows an exceptionally strong antioxidant capacity.
151
An investigation of the stem bark of Bersama abyssinica

Ian H. Bowen1,2, Betty P. Jackson1, Hemaia M. I. Motawe3
1 2 3
Pharmacognosy Research Laboratory, Sunderland Polytechnic, Sunderland, U. K. Department of Pharmaceutical Chemistry, Sunderland Polytechnic, Sunderland, U. K. Faculty of Pharmaceutical Sciences, Sunderland Polytechnic, Sunderland, U. K.
The barks of two subspecies of Bersama abyssinica, subspecies abyssinica and subspecies
-bufa-20,22-trienolide and 3-acetoxy-14-hydroxy-bufa-20,22-dienolide. Three sterols: -sitosterol, -stigmasten-3-ol and 4-methyl- -stigmast-dien-3-olhave also been isolated. 3 is a new compound
6 4 5,23
paullinioides have been shown to contain two bufadienolides: 5, 14-dihydroxy-3,6-diacetoxy-
and 3 and 4 are reported for the first time from the Melianthaceae. The presence of the xanthone mangiferin is a possible taxonomic marker for the genus within this family. Differences between the two subspecies were seen on examining the glycoside fractions. The alcoholic extracts did not exhibit any antitumour activity in the standard test.
Biosynthesis of mangiferin in anemarrhena incorporation of C6-C3 precursor into xanthone

Masao Fujita and Takao Inoue Hoshi College of Pharmacy,Shinagawa-ku,Tokyo,Japan
asphodewides:
intact
Mangiferin is a C-glycosylxanthone which is widely distributed in several families. It has been suggested that naturally occurring xanthones are biosynthesized via benzophenone-like intermediate derived wholly from polyketide in fungi, and from shikimate-polyketide in higher plants. The experiments indicate that the aglycone can be biosynthesized by the cyclization of an intermediate derived from p-coumarate and two malonates. This conclusion presents the occurrence of a new route for xanthone biosynthesis which is different from that of the xanthones in Gentiana lutea. The experiments strongly support that 3-C-glucosyl-maclurin would be a key intermediate for the biosynthesis of the above C-glucosylxanthones. Probable biosynthetic route of mangiferin and related C-glucosylxanthonesis shown in Scheme.
152
Study On Mango Leaf and Mangiferin Antiosteoporotic chemical constituents traditional Chinese herbal formula from Er-Xian Decoction,a
Luping Qin1, Ting Han1, Qiaoyan Zhang1, Dapeng Cao2, Hua Nian1,Khalid Rahman3, Hanchen Zheng1
1 2 3
School of Pharmacy, Second Military Medical University, Shanghai, P.R. China College of Chinese Medicinal Materials, Jilin Agricultural University, Changchun, P.R. China School of Biomolecular Science, Faculty of Science, Liverpool John Moores University, Liverpool, U.K.
Er Xian Decoction (EXD), a traditional Chinese medicine formula, has long been used for the treatment of osteoporosis and menopausal syndrome in China. The present study was designed to investigate the antiosteoporotic constituents of EXD, and evaluate their antiosteoporotic effects in ovariectomized rats. Osteoblasts in neonatal calvaria cultures and osteoclasts derived fromrat marrow cells were used to bioactivity-guided screen the active constituents. The proliferation of osteoblast was assayed by MTT methods. The activity of ALP and TRAP was measured by p-nitrophenyl sodium phosphate assay. The antiosteoporotic effects of icariin, anemarsaponin B II and berberine were verified by using OVX rats model. The bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry using the small animal scan mode. The undecalcified longitudinal proximal tibial metaphysical (PTM) sections were cut and stained for the bone histomorphometric analysis. Results: Bioactivity-guided fractionation has led to the successful isolation of antiosteoporotic constituents, i.e., icariin, icariside I, baohuoside I, mangiferin, neomangiferin, berberine, anemarsaponin B, anemarsaponin BII, anemarsaponin C, anemarrhenasaponin I, rubiadin- 1-methyl etherand obaculactone from EXD. Further study showed that icariin, anemarsaponin BII and berberine increased the BMD in ovariectomized rats, and icariin not only increased the bone formation, but also inhibited bone resorption; anemarsaponin BII mainly increased bone formation and berberine only inhibited the bone resorption in ovariectomized rats. These findings demonstrate that multiple ingredients are responsible for antiosteoporotic activity in traditional Chinese medicine formula Er-Xian decoction.
153
Study On Mango Leaf and Mangiferin Antioxidant C-Glucosylxanthones from the Leaves of Arrabidaea patellifer
Frdric Martin1, Anne-Emmanuelle Hay1, Delphine Cressend2, Marianne Reist2, Livia Vivas3, Mahabir P. Gupta4, Pierre-Alain Carrupt2, Kurt Hostettmann*,1
1
Laboratory of Pharmacognosy and Phytochemistry, School of Pharmaceutical Sciences, University of
Geneva, University of Lausanne, Switzerland

2 3
Unit of Pharmacochemistry, School of Pharmaceutical Sciences, University of Geneva, Switzerland University of Lausanne, Switzerland, London School of Hygiene and Tropical Medicine, Department of
Infectious and Tropical Diseases, London, U.K.

4
Center for Pharmacognostic Research on Panamanian Flora (CIFLORPAN), College of Pharmacy,
University of Panama, Panama
Chemical investigation of the methanol extract from the leaves of Arrabidaea patellifera, a Bignoniaceae from Panama, afforded mangiferin, isomangiferin, and six new derivatives mangiferin,
(3-O-p-hydroxybenzoylmangiferin,3-O-trans-coumaroylmangiferin,6-O-trans-coumaroyl-
3-O-trans-cinnamoylmangiferin, 3-O-trans-caffeoylmangiferin, and 3-O-benzoyl- mangiferin). All these compounds had antioxidant and radical-scavenging activities, and four of them were relatively active in vitro against Plasmodium falciparum. The structures were determined by spectrometric and chemical methods, including 1D and 2D NMR experiments and MS analysis.
A Xanthone C-glycoside from Iris Nigricans

Al-Khalil*,1, Hideki Tosa2, Munekazu Iinuma2
1 2
Faculty of Pharmacy, Jordan University for Women, Amman, Jordan Gifu Pharmaceutical University, Gifu, Japan
The isolation and identification of seven xanthones from an extract of the rhizomes of Iris nigricans is described. The isolated compounds are the xanthones, 1,3,6,7-tetrahydroxy- l,6,8-trihydroxy-2-methoxyl,6-dih ydrox y-3,7-dime thoxy-and 1,3,5,8-tetrahydrox y-,and the xanthone C-glycosides mangiferin,7-O-methylmangiferin and a new compound 2--D-glucopyranosyl-l,3,5,8-tetrahydroxyxanthone (nigricanside). The new structure was established by detailed spectral and chemical analysis.
154
Study On Mango Leaf and Mangiferin Benzophenone glycosides from Gnidia involucrate
Julien Ferrari1, Christian Terreaux1, Sevsen Sahpaz1, Jerome D. Msonthi2, Jean-Luc Wolfender1, Kurt Hostettmann1,*
1 2
Institut de Pharmacognosie et Phytochimie, Universit de Lausanne, Lausanne, Switzerland Department of Chemistry, University of Swaziland, Kwaluseni, Swaziland
Six compounds have been isolated from the methanol extract of the aerial parts of Gnidia involucrata (Thymelaeaceae). They were identified as 2,3,4',5,6-pentahydroxy-benzophenone -4-C-glucoside and 2,4',6-trihydroxy-4-methoxybenzophenone-2-O-glucoside, together with mangiferin, kaempferol-3-O-glucoside, yuankanin and manniflavanone by chemical and spectroscopic means. The structures of three additional C-glycosyl flavones-vitexin, isovitexin and isoorientin were determined on-line by LC/UV/APCI-MSn analysis of the crude extract.
Capillary electrophoresis analysis of mangiferin extracted from Mangifera indica L. bark and Mangifera persiciformis C.Y. Wu et T.L. Ming leaves
Chaozan Nong1, Weisheng He1, Debbi Fleming 2, Lili Pan1, Huayi Huang1 1 Department of Experimental Center, Guangxi Nationalities Hospital, Nanning, P.R. China 2 Department of Clinical Nutrition, Kaleida Health, Buffalo, New York, USA
The flavonoid compound mangiferin is found in the leaves, stem bark, fruit peels and root of Mangifera indica L. and in many other herbal species with many potential pharmacological properties. We have established an analytical method of mangiferin extracted from M. indica L. bark and Mangifera persiciformis C.Y.Wu et T.L. Ming leaves utilizing CZE. An electrolytic buffer containing 0.05M borate buffer, pH 7.4 with methanol(1:0.3, v/v) was deemed suitable for mangiferin analysis. An ideal mangiferin electropherogram with a migration time at pproximately 10.50 minwas obtained. Repeatability tests showed that the R.S.D.s for both intra- and inter-day migration time and peak area for all manigferin sourcestested were less than 4%. The linearity range of this method was 5-1000g/ml. The detection limit of this method was 1.5 g/ml. Quantitativeanalysis of mangiferin was also performed with this method. The accuracy of quantitation at 10, 500 and 1000g/ml of control mangiferin were99.00, 99.38 and 99.14%, respectively (n = 10). The repeatability of quantitation (R.S.D.) was below 3%. Our results demonstrated that CZE is asimple and reliable method in mangiferin analysis and more studies are needed to detect other mangiferin resources, such as clinical biological samples, in pharmacology and pharmacokinetic studies.
155
Study On Mango Leaf and Mangiferin Characterizaton of antioxidant and antiglycation properties and isolation of active ingredients from traditional Chinese medicines
Soon Yew Tang, Matthew Whiteman, Zhao Feng Peng, Andrew Jenner, Eu Leong Yong, and Barry Halliwell Department of Biochemistry, Faculty of Medicine, National University of Singapore, Singapore
There is considerable interest in the isolation of more potent antioxidant compounds to treat diseases involving oxidative stress. Thirty-three traditional Chinese medicine (TCM) extracts were examined for their antioxidant activity using the 2,2'-azinobis[3-ethylbenzothiazoline-6-sulfonate] (ABTS) assay. Five extracts with high activity (Cratoxylum cochinchinense, Cortex magnoliae officinalis, Psoralea corylifolia L, Curculigo orchioides Gaertn, and Glycyrrhiza uralensis Fisch.) were selected for further characterization. C. cochinchinense outperformed other extracts in most of the assays tested except phospholipid peroxidation inhibition, where P. corylifolia L showed higher activity. C. cochinchinense was particularly potent in inhibiting the formation of advanced glycation end products on proteins and strongly inhibited hypochlorous acid-induced DNA damage. We attempted to isolate the active ingredients from C. cochinchinense and obtained an extract (YCT) containing at least 90% mangiferin as identified by HPLC and mass spectrometry. However, YCT showed significantly higher activity in assays of phospholipid peroxidation, inhibition of protein glycation, and superoxide (O2-) and peroxynitrite (ONOO-) scavenging, as compared with mangiferin, suggesting that the nonmangiferin constituents of YCT contribute to its additional antioxidant activities.
Chemical constituents from Mahkota dewa

Bingzhang Yan, Junxu Xiang, Minliu Hong* New Drug Research and Development Centre, Zhengzhou University, Zhengzhou, China
A new phenolic glycoside , mahkoside A, together with six known compounds including mangiferin, kaempferol-3-O--D-glucoside , dodecanoic acid, palmitic acid, ethyl stearate and sucrose, were isolated from the pit of Mahkota dewa. Their structures were identified on the basis of spectroscopic analysis. All the compounds were isolated from the title plant for the first time.
156
Study On Mango Leaf and Mangiferin Characterization of polyphenols in mango puree concentrate by HPLC with diode array and mass spectrometric detection
Andreas Schieber, Wieland Ullrich, Reinhold Carle Hohenheim University,Institute of Food Technology,Section Plant Foodstuff Technology, Stuttgart, Germany
Polyphenols from mango puree concentrate were characterized by HPLC with diode array and mass spectrometric detection. After extraction with acetone, further fractionation of polyphenols with ethyl acetate and Sephadex LH-20 was necessary to obtain pure peaks. Five quercetin (Q) glycosides and one kaempferol glycoside were unambiguously identified. The predominant flavonol glycosides were Q 3-galactoside (22.1 mg/kg fresh wt.), Q 3-glucoside (16 mg/kg), and Q 3-arabinoside (5mg/kg). Among the phenolic acids, gallic acid was predominant(6.9mg/kg). Quantification of the C-glycoside mangiferin (4.4mg/kg) was also achieved by the HPLC method described. Using MS/MS, a gallotannin consisting of glucose and four gallic acid units was detected. Due to the presence of both carotenoids and polyphenols, mangos can be considered as an especially rich source of antioxidants.
Characterization of the mangiferin-human serum albumin complex by spectroscopic and molecular modeling approaches
Yuanyuan Yue, Xingguo Chen, Jin Qin, Xiaojun Yao Department of Chemistry, Lanzhou University, Lanzhou, P.R. China
The interactions between mangiferin andhumanserum albumin (HSA) were investigated by spectroscopy and molecular modeling. The results proved the formation of complex between mangiferin and HSA. Hydrophobic interaction dominated in the association reaction. Mangiferin statically quenched the fluorescence of HSA in a concentration dependent manner positively deviating from the linear Scatchard equation. The binding of mangiferin to HSA lead to changes in the conformation of HSA according to synchronous fluorescence spectra, FT-IR, UVvis and CD data. The presence of amino acids and metal ion affected the binding constant of mangiferinHSA complex. Computational mapping of the possible binding sites of mangiferin revealed the molecule to be bound in the large hydrophobic cavity of subdomain IIA.
157
Study On Mango Leaf and Mangiferin Chemical and chemotaxonomical studies of ferns. LXXXVII. constiuents of trichomanes reniforme
Hiroshi Wada1,Yasufumi Shimizu1, Nobutoshi Tanaka1, Richard C. Cambie2,John E. Braggins3
1 2 3
Faculty of Pharmaceutical Sciences, Science University of Tokyo, Tokyo,Japan Department of Chemistry,The University of Auckland, New Zealand Department of Botany,The University of Auckland,New Zealand
Five
new
glycosides,3,4-dihydroxyphenethyl
alcohol
4-O-caffeoyl--D-allopyranoside,
(6S,
13S)-13--D-fucopyranosyl-6-{-D-fucopyranosyl-(1-2)-[-D-fucopyranosyl-(1-4)--L-rhamno-pyranosyloxy]}-cleroda-3,14-diene, (6S,13S)-13- -D- fucopyranosyloxy-6-{-D-quinovo-pyranosyl-(1-2)-[-Dfucopyranosyl-(1-4)--L-rhamnopyranosyloxy]}-cleroda-3,14-diene, (6S,13S) -13--L-arabinopyranosyloxy-6-{-D-fucopyranosyl-(1-2)-[ -D-fucopyranosyl- 1-4
--L- rhamnopyranosyloxy]}-cleroda-3,14-
diene and (6S,13S)-13--L- arabinopyranosyloxy -6-{-D-quinovopyranosyl-(1-2)-[ -D- fucopyranosyl(1-4)--L-rhamnopyranosyloxy]}-cleroda-3,14-diene,were isolated,together with mangiferin and 6'-Oacetylmangiferin,from the fronds of a New Zealand fern, Trichomannes reniforme.
Chemical constituents of Gentianaceae XIX: CNS-depressant effects of swertiamarin

S. K. Bhattacharya1, P. K. S. P. Reddy1, S. Ghosal2, A. K. Singh2, P. V. Sharma2
1 2
Merrell-Nationul Laboratories, Division of Richardson-Merrell Inc. Cincinnati, USA Section of Medicinal Chemistry and Pharmacognosy, School of Pharmacy, University of Connecticut,
Storrs, USA
CNS activity of swertiamarin, a secoiridoid glucoside from Swertia chirata, was evaluated. An apparent anomaly, associated with the unanticipated finding that the alcoholic extracts (excluding mangiferin) of S. chirata significantly reversed the marigiferin-induced CNS-stimulating effects in albino mice and rats, was resolved. The results indicate that swertiamarin and mangiferin antagonize each other in vivo and thereby reverse their CNS effects.
158
Study On Mango Leaf and Mangiferin Determination of gentiopicroside, mangiferin, palmatine, berberine, baicalin, wogonin and glycyrrhizin in the traditional Chinese medicinal preparation Sann-Joong-Kuey-Jian-Tang by high- performance liquid chromatography
Shion-Jane Lin, Hshinn-Hshiung Tseng, Kuo-Ching Wen*, Tsi-Tee Suen National Laboratories of Foods and Drugs, Department of Health, Executive Yuan, Nankang, Taipei, Taiwan
High-performance liquid chromatography was employed to determine the contents of several marker substances such as gentiopicroside, mangiferin, palmatine, berberine, baicalin, wogonin and glycyrrhizin in Sann-Joong-Kuey-Jian-Tang. The separation was performed on a Cosmosil 5C18-AR column by gradient elution with 0.03% (v/v)phosphoric acid-acetonitrile (0 min, 90:l0; 10 min, 87:13; 17-27 min, 77:23; 40 min, 62:38; 50 min, 5545) as the mobile phase at a flow-rate of 1.0 ml/min, with detection at 254 nm. n-Propylparaben was used as the internal standard and seven regression equations revealed linear relationships between the peak-area ratios (marker substances/internal standard) and concentrations. The repeatability and reproducibility (relative standard deviation) of the method were in the ranges 0.02-1.78% and 1.44-4.95%, respectively.
Determination of the residue of organochlorine pesticides in mango leaves using GC-MS-SIM

Jieping Qin, Jiagang Deng, Yunyun zhang , Xu Feng, Zedong Chen Guangxi Traditional Chinese Medical University, Nanning, PR China
A simple and fast method has been developed to determine 9 organic chlorine pesticides in 10 batches of mango leaves. The results indicated this method had good relativity of linearity in 0.01ng-0.1ng, and the limits of detection were lower than 0.05ng for gas chromatography with electron impact mass spectrometric detection in the selected ion monitoring mode (GC-MS-SIM). The recovery results found ranged from 96.75 % to 101.84 % and RSD% was less than 3.73. The determination results shown that the residues of organochlorine pesticides in 10 batches of mango leaves which collected from different areas were all less than 210-7g/g.
159
Study On Mango Leaf and Mangiferin Determination of mangiferin, jateorrhizine, palmatine, berberine, cinnamic Acid, and cinnamaldehyde in the traditional Chinese medicinal preparation Zi-Shen Pill by high-performance liquid chromatography
Ronghua Dai, Kang Li, Qing Li, and Kaishun Bi* Shenyang Pharmaceutical University, Wenhua Road 103, Shenyang, P.R. China
High-performance liquid chromatography is employed to determine the contents of six marker components such as mangiferin,jateorrhizine, palmatine, berberine, cinnamic acid, and cinnamaldehyde in the traditional Chinese medicinal preparation Zi-Shen pill. The separation is performed on a C18 column by stepwise gradient elution with water (0.2%, v/v, triethylamine adjusted to pH 4 with phosphoric acid)-methanol-acetonitrile (0.01min, 98:0:2; 20 min, 80:5:15; 30 min, 65:13:22; and 55 min,65:13:22) as the mobile phase at a flow rate of 0.9 mL/min, with UV detection at 280 nm. Six regression equations show good linear relationships between the peak area of each marker and concentration. The recoveries of the markers listed are 95.5%, 98.3%, 96.8%, 99.5%, 101.7%, and 102.1%, respectively. The repeatability and reproducibility (relative standard deviation) of the method are less than 2.5% and 3.3%, respectively.
Differentiation of Swertia Mussotii Franch from Artemisiae Capillaris Herba by capillary electrophoresis with electrochemical detection
Yuhua Cao1, Yun Wang1, Jiannong Ye2
1 2
School of Chemical and Material Engineering, Southern Yangtze University, Wuxi, PR China Department of Chemistry, East China Normal University, Shanghai, PR China
A high-performance capillary electrophoresis (CE) with electrochemical detection (ED) method is developed for differentiation of Swertia Mussotii Franch from Artemisiae Capillaris Herba in this work. Swertia Mussotii Franch contains a great deal of swertiamarin and mangiferin that are not present in Artemisiae Capillaris Herba, whereas Artemisiae Capillaris Herba consists of abundant chlorogentic acid. Therefore, determining their swertiamarin, mangiferin and chlorogentic acid contents can differentiate these two crude herbs. Operated in a wall-jet configuration, a 300m diameter carbon-disk electrode was used as the working electrode, which exhibits good response at +1000mV (versus SCE) for the three analytes. With a separation voltage of 14 kV, the three analytes were separated within 14 min in a 52 cm length capillary in 50 mmol/l borax buffer (pH 9.2). The system was demonstrated good stability and reproducibility with an R.S.D. of less than 5% for both migration time and peak current. This method was successfully used to analyze and identify the crude herbs with satisfactory assay results.
160
Study On Mango Leaf and Mangiferin Evaluation of spectrophotometric methods for screening of green rooibos (Aspalathus linearis) and green honeybush (Cyclopia genistoides) extracts for high levels of Bio-active compounds
Elizabeth Joubert1,2,Marena Manley2,Mariza Botha2
1 2
ARC Infruitec-Nietvoorbij, Stellenbosch, South Africa Department of Food Science, Stellenbosch University, Matieland (Stellenbosch), South Africa
The potential of UV spectrophotometry and an aluminium chloride (AlCl3) colorimetric method to determine the dihydrochalcone (DHC) and mangiferin contents of green rooibos and honeybush (C. genistoides) extracts, respectively, was investigated.The DHC content of rooibos water extracts, determined using UV spectroscopy, correlated with the sum of the aspalathin and nothofagin contents as quantified using HPLC (r=0.98). A correlation coefficient of 0.91 was obtained when correlating the mangiferin content of C. genistoides methanol extracts, determined by the AlCl3 colorimetric method, with the results obtained by HPLC. Using the linear equations from the correlations it was possible to predict the DHC and mangiferin contents of extracts from the respective spectrophotometric measurements to a reasonable accuracy as an alternative to HPLC. The total polyphenol(TP) content of rooibos water extracts can also be determined using UV spectrophotometry and aspalathin as a standard (r=0.99) as an alternative to the Folin-Ciocalteau method. The TP content of rooibos extracts correlated (r=0.99) with its total antioxidant activity (TAA) as determined with the ABTS radical cation scavenging assay, but the TP content of C. genistoides water extracts is not a good indication of their TAA (r=0.27). The aspalathin content of rooibos extracts correlated with their TAA (r =0.96), but the mangiferin content of honeybush water extracts only gave a moderate correlation with their TAA(r=0.75).
161
Study On Mango Leaf and Mangiferin Flavonoid and xanthone patterns in bearded Iris species and the pathway of chemical evolution in the genus
Christine A. Williams1, Jeffrey B. Harborne1, Maretta Colasante2
1 2
Department of Botany, University of Reading, U.K. Department of Plant Biology, University La Sapienza, Rome, Italy
Flavonoids, xanthones and isoflavones have been surveyed in leaf, flower and rhizome of four diploid and seven allopolyploid taxa of Bearded Iris, in order to investigate their phylogeny. Eighteen glycoflavones and eight glucoxanthones were identified in the leaves and these compounds proved to be the most useful evolutionary markers. Both the diploid and allopolyploid taxa showed stable patterns, with minimal variation below the species level. The results showed that the species Iris pseudopumila was a strong candidate as a diploid parent, while they ruled out the participation of Iris pallida. Of the allopolyploids, I. germanica and I.albicans appeared to be very close, and I. biflora was also closely related possibly because of their supposed hybrid origin, cultivation and successive escapes from it. Iris marsica, which is endemic to the Abruzzi Region of Italy, has a distinct phenolic profile. Anthocyanin patterns in the flowers and isoflavone patterns in the rhizomes were generally uniform throughout the group.
162
Study On Mango Leaf and Mangiferin Glucuronide triterpene saponins from Bersama engleriana
Azefack Lon Tapondjou1,3, Tomofumi Miyamoto2, Marie-Aleth Lacaille-Dubois1,*
1
Laboratoire de Pharmacognosie, Unit de Molcules dIntrt Biologique, UMIB UPRES EA3660,
Facult de Pharmacie, Universit de Bourgogne, Dijon Cedex, France

2 3
Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan Laboratoire de Chimie Applique et Environnementale, Facult des Sciences, Universit de Dschang,
Dschang, Cameroon
Five 3-O-glucuronide triterpene saponins were isolated from the stem bark of Bersama engleriana Gurke along with two known saponins, polyscias saponin C and aralia saponin 15, and one major C-glycoside xanthone, mangiferin. The structures of the saponins were established mainly by means of spectroscopic methods (one- and two-dimensional NMR spectroscopy as well as FAB-, HRESI-mass spectrometry) as 3-O-[-D-glucopyranosyl-(12)--D-glucuronopyranosyl]-28-O-[- D- glucopyranosyl] -betulinic acid, 3-O- [- D-glucopyranosyl -(12)- [ -D-galactopyranosyl-(13)]- -D-glucurono
pyranosyl]-oleanolic acid , 3-O-[-D-glucopyranosyl-(13)--D-glucuronopyranosyl]-28-O-[- D-xylopyranosyl-(16)--D-glucopyranosyl]-oleanolic acid, 3-O-[-D-galactopyranosyl-(13)- -D-glucuronopyranosyl]-28-O-[-D-glucopyranosyl-(14)- -D-glucopyranosyl]-oleanolic acid , and 3-O-[-D-glucopyranosyl-(13)--D-galactopyranosyl-(13)--D-glucuronopyranosyl]-28-O-[-D- xylopyranosyl(16)- -D-glucopyranosyl]-oleanolic acid.
163
Study On Mango Leaf and Mangiferin High-performance liquid chromatographic method for the determination and pharmacokinetic study of mangiferin in plasma of rats having taken the traditional Chinese medicinal preparation Zi-Shen pill
Ronghua Dai*, Jun Gao, Kaishun Bi Shenyang Pharmaceutical University, Shenyang, P.R. China
A high-performance liquid chromatographic method for the determination and pharmacokinetic study of mangiferin in the plasma of rats that have been orally administered the traditional Chinese medicinal preparation Zi-Shen pill is established. Plasma samples taken from rats are pretreated by protein precipitation with acetonitrile. Separation of the main effective constituent mangiferin is accomplished on a C18 stationary phase and a mobile phase of methanol-water (25:75, v/v) with 0.6% glacial acetic acid. The UV detection wavelength is set at 320 nm, and the detection limit for mangiferin in plasma is 0.163 g/mL. After validation, the method is used to take a limited view of pharmacokinetic profiles of the traditional Chinese medicinal preparation Zi-Shen pill.
High-performance liquid chromatography as a tool for the chemical standardisation of triphala-an ayurvedic formulation
D. P. Singh, R. Govindarajan, A. K. S. Rawat* Pharmacognosy and Ethnopharmacology Division, National Botanical Research Institute, Lucknow, India
Triphala is an anti-oxidant-rich herbal formulation containing fruits of Emblica officinalis, Terminalia chebula and T. belerica in equal proportions. The preparation is frequently used in Ayurvedic medicine to treat diseases such as anaemia, jaundice, constipation, asthma, fever and chronic ulcers. Anti-mutagenic effects of the polyphenolic fractions isolated from Triphala have been reported, thus indicating that the phenols present in the formulation might be responsible for its therapeutic efficacy. A simple high-performance liquid chromatography method for the separation and quantitative determination of the major antioxidant polyphenols from Triphala has been developed. The use of an RP18 column with an acidic mobile phase enabled the efficient separation of gallic acid, tannic acid, syringic acid and epicatechin along with ascorbic acid within a 20 min analysis. Validation of the method was performed in order to demonstrate its selectivity, linearity, precision, accuracy and robustness. In addition, optimisation of the complete extraction of phenolic compounds was also studied.
164
Study On Mango Leaf and Mangiferin Isolation of isomangiferin from honeybush (Cyclopia subternata) using high-speed counter-current chromatography and high performance liquid chromatography
Dalene de Beer 1,*, Gerold Jerz 2, Elizabeth Joubert 1,3, Victor Wray 4, Peter Winterhalter 2
1 2 3 4
Post-Harvest and Wine Technology Division, ARC Infruitec-Nietvoorbij, Stellenbosch, South Africa Institute of Food Chemistry, Technical University of Braunschweig, Braunschweig, Germany Department of Food Science, Stellenbosch University, Stellenbosch, South Africa Department of Structural Biology, Helmholtz Centre for Infection Research, Braunschweig, Germany
Isomangiferin was isolated from Cyclopia subternata using a multi-step process including extraction, liquid-liquid partitioning, high-speed counter-current chromatography (HSCCC) and semi-preparative reversed-phase HPLC. Enrichment of phenolic compounds in a methanol extract of Cyclopia subternata leaves was conducted using liquid-liquid partitioning with ethyl acetate-methanol-water (1:1:2, v/v). The enriched fraction was further fractionated using HSCCC with a ternary solvent system consisting of tert-butyl methyl ether-n-butanol-acetonitrile-water (3:1:1:5, v/v). Isomangiferin was isolated by semipreparative reversed-phase HPLC from a fraction containing mostly mangiferin and isomangiferin. The chemical structure of isomangiferin was confirmed by LC high-resolution electrospray ionization MS, as well as one and two dimensional NMR spectroscopy.
165
Study On Mango Leaf and Mangiferin Isolation of Mangiferin and Isomangiferin from Leaf Material of Hibiscus liliastrum (Malvaceae)
Stephen Cafferty, Jenny Greenham, Christine A. Williams* Department of Botany, Plant Sciences Laboratories, The University of Reading, Reading, U.K.
Leaf material (0.2 g) from the herbarium specimens was extracted with 70% ethanol and the phenolic constituents isolated from the concentrated extracts by multiple two-dimensional chromatography. The mangiferin (1) and isomangiferin (2) spots (both dark orange to yellow in UV light and ammonia) were cut out, and eluted in 80% methanol. The concentrated eluates were run on a Waters 600 HPLC with photo-diode array detector (940) using a gradient method and a reverse phase Bondapak phenyl column. The proportions of Solvent A (2% HOAc) to Solvent B (MeOH:HOAc:H2O: 18:l:l) were changed from 75% A/25% B to 100% B over 20 min in linear mode at a flow rate of 1ml/l min at room temperature with detection at 260 nm. The retention time of each sample was compared with that of an authentic mangiferin marker. The identity of 1 was confirmed as mangiferin by co-chromatography on cellulose TLC in BAW. 15% HOAc, H2O and CAW (CHCl3:HOAc, 1:1 saturated with H2O), and from UV spectral data. Compound 2 was identified as isomangiferin (the corresponding-4-C-glucoside) from its Rf and Rt values, and from UV spectral data compared with those of mangiferin and relevant literature data.
Isolation of mangiferin from Bombax malabaricum and structure revision of shamimin

Abdelaaty A. Shahat1,2, Rasmeria A. Hassan1, Naglaa M. Nazif1, Sabine Van Miert2, Luc Pieters2, Faiza M. Hammuda1, Arnold J. Vlietinck2
1 2
Department of Pharmaceutical Sciences, National Research Centre, Dokki, Cairo, Egypt Department of Pharmaceutical Sciences, University of Antwerp, Antwerp, Belgium
Repeated column chromatography of the n-BuOH fraction of the 70% EtOH of the dried leaves of Bombax malabaricum led to the isolation of mangiferin, a xanthone. Mangiferin was identified by UV,
1
H-and 13C-NMR spectroscopy and electrospray mass spectrometry. It was found to be identical to
shamimin, a compound for which originally a flavonol structure was proposed, and the structure of which has to be revised.
166
Study On Mango Leaf and Mangiferin Mangiferin and isomangiferin in some Hypericum species
Gerassim M. Kitanov*, Paraskev T. Nedialkov Department of Pharmacognosy, Faculty of Pharmacy, Medical University of Sofia, Bulgaria
Mangiferin was found in 26 and isomangiferin in 33 of the 36 evaluated species from Hypericum L., belonging to 17 sections of the genus. These substances are reported here in 25 taxa for the first time. In most of the investigated species they were minor components. The highest amounts of mangiferin were observed in H. rochelii Griseb. et Schenk, H. perfoliatum L., H. aucheri Jaub. et Spach and H. momtanum L. The widely distribution of mangiferin and isomangiferin in species of Hypericum appear to be little chemotaxonmic significance for the sectional classification of the genus. These xanthone C-glucosides seem to possess value as taxonomic markers at the subfamily level. At present they are characteristic only for the taxa of subfamily Hypericoideae and have not been found in the remaining subfamilies of the Guttiferae.
Mangiferin Identified in a Screening Study Guided by Neuraminidase Inhibitory Activity

Xiaofan Li1, Takashi Ohtsuki1, Sayaka Shindo1, Masaaki Sato1, Takashi Koyano2, Srisomporn Preeprame3, Thaworn Kowithayakorn4,Masami Ishibashi1
1 2 3 4
Graduate School of Pharmaceutical Sciences, Chiba University, Chiba,Japan. Temko Corporation, Tokyo, Japan. Faculty of Pharmaceutica Sciences, Khon Kaen University, Khon Kaen,Thailand. Faculty of Agriculture, Khon Kaen University, Khon Kaen,Thailand
A screening study on neuraminidase inhibitory constituents was carried out,and activity-guided fractionations of three plants,Gouania obtusifolia,Zizyphus cambodiana,and Mangifera odorata, led to the isolation of eleven compounds.Mangiferin was identified as a significant neuraminidase inhibitor.
167
Study On Mango Leaf and Mangiferin Liquid chromatography/tandem mass spectrometric study and analysis of xanthone and secoiridoid glycoside composition of Swertia chirata, a potent antidiabetic
Satyendra Suryawanshi1,Nitin Mehrotra1,R.K.Asthana2 and Ram C.Gupta1*
1 2
Pharmacokinetics and Metabolism Division,Central Drug Research Institute,Lucknow,India Medicinal and Process Chemistry Division,Central Drug Research Institute,Lucknow,India
Swertia chirata is a bitter plant,used in the Indian system of medicine(Ayurveda)for various human ailments.The bioactive constituents include the xanthone and secoiridoid glycosides consisting of mangiferin,amarogentin,amaroswerin,sweroside and swertiamarin.Methanolic extracts of S.chirata possess constituents with antidiabetic activities,which was investigated by highperformance liquid
chromatography/electrospray ionization tandem mass spectrometry(LC/ESIMS/MS).Preliminary HPLC analyses were performed on a reversed-phase C18 column using gradient elution.In the LC/ESI-MS spectra,predominant[M+H]
+
and[M+Na]
ions were observed in positive ion mode and provided
molecular mass information.The five components of S.chirata were structurally correlated and confirmed based on the fragmentation characteristics and information available in the literature.The fragmentation behavior of [M+H] +/[M+Na]+ ions of these components were deduced from the collision-induced dissociation(CID)spectra obtained from the selective on-column information-dependant
acquisition(IDA)approach.Xanthone-C-glycoside showed characteristic fragment ions due to fragmentation in the C-glycosidic unit while iridoid-O-glycosides showed characteristic fragment ions due to cleavage in the glycoside linkage and retro-Diels-Alder(RDA)cleavage within an iridoid aglycone.Furthermore,on the basis of this information,an analytical assay was developed and validated to determine relative concentrations of mangiferin, amarogentin, amaroswerin, sweroside and swertiamarin.The detection was carried out using multiple reaction monitoring (MRM) in positive ionization mode with a total analysis time of 3.5 min.The method was successfully applied to standardize four different batches of herbal preparation on the basis of relative concentration of five bioactive components.
168
Study On Mango Leaf and Mangiferin Mangeiferin from the root bark of salaczaretzculata
E.H. Karunanayake and S.R. Sirimanne Department of Biochemistry, Faculty of Medicine, University of Colombo, Colombo 8 (Sri Lanka)
According to the Ayurvedic system of medicine practised in Sri Lanka,several plants have been claimed to possess oral hypoglycaemic activity(Attygale, 1917; Chandrasena, 1935). However, most of these plants have not been subjected to systematic chemical and pharmacological investigations. Salacia reticulatu (Celastraceae) is most commonly and widely used by the Ayurvedic practitioners in the treatment of diabetes mellitus. The mode of administration consists of an aqueous decoction prepared from the roots of the plant. We have recently reported (Karunanayake et al., 1984) the oral hypoglycaemic activity of this plant preparation in laboratory animals. In the course of our investigation of the oral hypoglycaemic principles of this plant, we have isolated mangiferin from its root bark.Powdered root bark (100 g) of Salucia reticuluta was boiled with distilled water (1L) for 3 h. The aqueous extract was cooled and freeze-dried. The lyophilized product was chromatographed on Sephadex LH-20 using 95% ethanol as the eluent. A pale yellow solid was obtained on evaporation of ethanol. Recrystallization from 75% ethanol gave mangiferin as pale yellow needles, m.p. 271-272C (found: C 54.32, H 4.45; C19H18011 requires C 54.0, H 4.3%). Paper chromatographic behaviour, colour of the spot under UVZ+, and UVZ5JNH3, m.p., IR, UV and NMR data were identical with the recorded data for authentic mangiferin (Billet et al., 1965; Haynes and Taylor, 1966, Bhatia et al., 1967). Attempted hydrolysis of mangiferin gave no sugar (Billet et al. 1965). Acetylation with acetic anhydridelpyridine gave mangiferin hepta-acetate, m.p. 229-230C. NMR spectrum of the hepta-acetate was identical with the recorded data for mangiferin in hepta-acetate (Billet et al., 1965).Investigation of a possible oral hypoglycaemic activity of mangiferin by the procedure previously reported (Karunanayake et al., 1984) showed no such activity in laboratory animals.Although mangiferin has been isolated from Salacia pirinoides (Pilay and Lekshuri, 1958), this is the first report of the occurrence of mangiferin in Salacia reticulata. The compound had no oral hypoglycaemic activity when tested in laboratory animals. This work was supported by grant No. 531 from the International Foundation for Science, Sweden and RG 81/23 from the Natural Resources, Energy and Science Authority of Sri Lanka. The authors are thankful to the Department of Chemistry, University of Peradeniya, Sri Lanka for providing facilities to obtain NMR and IR spectra.
169
Study On Mango Leaf and Mangiferin Miscibility Characterization in Relation to Phase Morphology of Poly (ether sulfone)/Poly (vinyl pyrrolidone) Blends Containing a Phytochemical
Chandrasekaran Neelakandan Thein Kyu* Department of Polymer Engineering,University of Akron,Akron,Ohio,U.S.A.
Miscibility and morphology of poly(ether sulfone)/poly(vinyl pyrrolidone)(PES/PVP)blends containing a crystalline phytochemical called mangiferin were investigated using differential scanning calorimetry(DSC),Fourier transformed infrared spectroscopy(FTIR),and polarized optical
microscopy(POM).The binary blends of PES/PVP were found to be completely miscible.However,FTIR experiments revealed no spectral shift that is attributable to the miscibility of the PES/PVP pair,although the occurrence of hydrogen-bonding interactions can be confirmed in binary blends of both PES/mangiferin and PVP/mangiferin.The addition of mangiferin to the PES/PVP blends resulted in liquid-liquid phase separation as well as liquid-solid phase transition.However,the liquid-liquid phase separation was observed only in a very small ternary composition range of the PES/PVP/mangiferin blends.With further increase of mangiferin concentration,crystallization occurred,leading to phase segregation between the isotropic liquid(PES/PVP)phase and the crystalline mangiferin.A ternary morphology phase diagram of the PES/PVP/mangiferin blends was established based on the evidence from DSC and POM experiments,which exhibited various coexistence regions including isotropic, liquid+liquid, liquid+crystal, and solid crystal regions.
New Steroidal Sapomins from the Rhizomes of Anemarrhena asphodeoides Bunge (Liliaceae)
Setsuo Saito*, Satoshi Nagase, Koki Ichinose Faculty of Pharmaceutical Sciences, Josai University,Keyakidai 1-1,Sakado,Saitama 350-02,Japan
From the rhizome of Anemarrhena asphodeoides Bunge(Liliaceae),four new steroidal saponins named anemarrhenasaponins - were isolated,together with known saponins, timosaponinA- ,marcogenin diglycoside and timosaponin B-
and a xanthone C-glycoside, mangiferin. These saponina are coprostane
type steroidal glycosides.Their structures were established on the basis of spectroscopic and chemical evidence.
170
Study On Mango Leaf and Mangiferin On-line purity monitoring in high-speed counter-current chromatography Application of HSCCC-HPLC-DAD for the preparation of 5-HMF, neomangiferin and mangiferin from Anemarrhena asphodeloides Bunge
Tingting Zhou1,Zhenyu Zhu1,Chen Wang1,Guorong Fan2,Jinyong Peng,Yifeng Chai,Yutian Wu
1 2
Shanghai Key Laboratory for Pharmaceutical Metabolite Research, Shanghai,P.R.China School of Pharmacy,Second Military Medical University, Shanghai,P.R.China
An efficient on-line purity monitoring strategy based on on-line coupling of high-speed counter-current chromatography(HSCCC)with highperformance liquid chromatography-diode array detection(HPLC-DAD)was successfully applied for the first time to the isolation and purification of 5-hydroxymethyl-furancarboxaldehyde(5-HMF),mangiferin and neomangiferin from the Chinese medicinal plant Anemarrhena asphodeloides Bunge,a plant used in the traditional Chinese medicine.The introduction of on-line purity monitoring in HSCCC has greatly improved the efficiency of this technique by overcoming the drawbacks of post-purification sample handling in HSCCC isolation.The effluent from the outlet of HSCCC was splitted into two parts,and one was collected,while the other was introduced directly through a switch valve into a HPLC-DAD system for purity monitoring.Using this method the desired fractions with high purities could be collected.From 600 mg partially purified extract,165.6 mg neomangiferin and 292.8 mg mangiferin with purities of 98.9 and 99.5%,respectively,were obtained with a two-phase solvent system composed of n-butanolwater(1:1,v/v)by increasing the flow-rate of the mobile phase stepwise from 1.0 to 2.2 ml min-1 after 210 min.A 17.1 mg 5-HMF with purity of 96.6%was also isolated for the first time.
171
Phenolic compounds from Hypericum perforatum

Guido Jrgenliemk1,2, Adolf Nahrstedt1
1
Institute of Pharmaceutical Biology and Phytochemistry,Westfalische Wilhelms-University of Mnster,
Germany
2
Part of the PhD thesis of G Jrgenliemk,Germany
During a re-investigation of phenolic compounds from the dried crude drug material of St.John's wort(Hypericum perforatum L.) 22 phenolic compounds were detected by HPLC;14 of them were quantified using the same system..Twelve phenolic compounds were isolated from the plant material and their structures identified mainly by spectroscopic methods,among them quercetin-3-O-(2-O-acetyl)--Dgalactoside as a new natural product.Cryptochlorogenic acid,protocatechuic acid,3-O-[Z]-p-coumaroyl -quinic acid,isoorientin,cyanidin-3-O- -L-rhamnoside,and astilbin were obtained for the first time from this source;the earlier suspected neochlorogenic acid,3-O-[E]-p-coumaroylquinic acid,mangiferin, miquelianin and guaijaverin were confirmed.
Phenolic metabolites from honeybush tea (cyclopia subternata)

B.Irene Kamara*1,D.Jacobus Brand1,E.Vincent Brandt*1,Elizabeth Joubert2
1 2
Department of Chemistry, University of the Free State,Bloemfontein, South Africa ARC Infruitec-Nietvoorbij, Stellenbosch, South Africa
Cyclopia subternata is one of the 24 Cyclopia species that are used to brew honeybush tea, a unique South African herbal beverage with a pleasant taste and flavor. It contains various antioxidants, very low tannin content, and no caffeine. Many health properties are associated with regular consumption of the tea. Honeybush infusions have been noted as a tonic for colds and influenza, catarrh, and pulmonic tuberculosis and is becoming well-known for its effectiveness in alleviating menopausal symptoms in women. Unfermented leaves of C. subternata contain pinitol, shikimic acid, p-coumaric acid, 4-glucosyltyrosol, epigallocatechin gallate, the isoflavone orobol, the flavanones hesperedin, narirutin, and eriocitrin, a glycosylated flavan, the flavones luteolin, 5-deoxyluteolin, and scolymoside, the xanthone mangiferin,and the flavonol C-6-glucosylkaempferol.The structures were elucidated by spectroscopic and spectrometric analysis.
172
Study On Mango Leaf and Mangiferin Polyphenol constituents from salacia species: quantitative analysis of mangiferin with a-glucosidase and aldose reductase inhibitory activities
Masayuki Yoshikawa*1, Norihisa Nishida1, Hiroshi Shimoda2, Miki Takada2, Yuzo Kawahara2, Hisashi Matsuda1
1 2
Kyoto Pharmaceutical University, Kyoto, Japan and Morishita; Jintan Co., Ltd., Osaka, Japan
Mangiferin, three catechins, and two catechin dimers were isolated from the roots of Salacia reticulata (SRE), and examined their inhibitory activities against several carbohydrate metabolize enzymes (sucrase, maltase, isomaltase, -amylase, and aldose reductase). Among them, mangiferin was found to inhibit sucrase, isomaltase, and aldose reduc-tase from rat with IC50 values of 87, 216 and 1.4 g/ml, respectively. The inhibitory activities of mangiferin are competitive for sucrase and isomaltase with inhibitor constant (Ki) 55 g/ml and 70 g/ml, respectively. In order to determine the mangiferin contents in the water extracts from the roots of S. reticulata , a quantitative analytical method by means of HPLC was developed and the mangiferin contents in SRE were determined to be in the range of 0.9-2.3 by the application of this method. A high linear correlation (r 0.934) was observed between the mangiferin contents and the sucrase inhibitory activity. In addition, this analytical procedure of mangiferin was found to be applicable for other Salacia species (S. oblonga, S. chinensis, and S. prinoides). Thus, the quantitative HPLC analysis of mangiferin was supposed to be suitable for the quality control of Salacia species and its products.
173
Study On Mango Leaf and Mangiferin Preparative isolation and purication of four compounds from the chinese medicinal herb rhizoma anemarrhenae by high-speed counter-current chromatography
Qinghua Sun, Ailing Sun, Renmin Liu* College of Chemistry and Chemical Engineering, Liaocheng University, Liaocheng, P.R.China
High-speed counter-current chromatography (HSCCC) was applied to the separation and purication of mangiferin, neomangiferin, cis-hinkiresinol and (-)-4/-O-methylnyasol from the Chinese medicinal herb Rhizoma Anemarrhenae. Five hundred milligrams of crude extracts were separated by using n -butanolacetic acid (1%) (1:1, v/v) as the two-phase solvent system and yielded 35.3mg of neomangiferin and 245.4mg of mangiferin. During this separation, cis -hinkiresinol and (-)-4/-O-methylnyasol were stillmaintained in the stationary phase. The stationary phase was collected, evaporated to dryness and separated with light petroleumethyl acetatemethanolwater (1:1:1.2:0.8, v/v) and 1:1:1.4:0.6 (v/v) in gradient elution, which yielded 17.2mg of cis-hinkiresinol and 12.4mg of (-)-4 / -O -methylnyasol. The purities of mangiferin, neomangiferin, cis -hinkiresinol and (-)-4 / -O methylnyasol were 96.3, 98.0, 97.3 and 98.2%, respectively, as determined by HPLC. The chemical structures of these components were identied by 1H NMR and 13C NMR.
Chemical constituents in the leaves of Mangifera persiciformis C.Y. Wu et Y.L. Ming

XiuLing Si, Song Wei, XueJiang Xu, Xuehuan Fang, WenJiang Wu
Department of Pharmacy, Guangxi Traditional Chinese Medical University, Nanning 530001
Eleven crystalline constituents have been isolated from the leaves of Mangifera persiciformis, of which five were identified as taraxerol, friedelin, beta-sitosterol, mangiferin and quercetin by comparing their physicochemical and spectroscopic data. They were isolated from this plant for the first time.
174
Study On Mango Leaf and Mangiferin Quality evaluation of rhizoma belamcandae (belamcanda chinensis (L.) DC.) by using high-performance liquid chromatography coupled with diode array detector and mass spectrometry
Jun Li, Winnie Z.M. Li, Wen Huang, AnnaW.H. Cheung, Cathy W.C. Bi, Ran Duan, Ava J.Y. Guo, Tina T.X. Dong, KarlW.K. Tsim Center for Chinese Medicine and Department of Biology, the Hong Kong University of Science and Technology, Hong Kong, P.R.China
A high-performance liquid chromatography coupled with diode array detector and mass spectrometry (HPLC-DAD-MS) method was developed to evaluate the quality of Rhizoma Belamcandae (Belamcanda chinensis (L.) DC.) through establishing chromatographic ngerprint and simultaneous determination of seven phenolic compounds. The analysis was achieved on an Alltima C18 analytical column (250mm4.6mm i.d. 5m) using linear gradient elution of acetonitrile0.1% triuoroacetic acid. The correlation coefcients of similaritywere determined fromthe HPLC ngerprints, and they shared a close similarity. By using an online APCI-MS/MS, twenty phenols were identied. In addition, seven of these phenols including mangiferin, 7-O-methylmangiferin, tectoridin,resveratrol, tectorigenin,irigenin and irisorentin were quantied by the validated HPLC-DAD method. These phenols are considered to be major constituents in Rhizoma Belamcandae, and are generally regarded as the index for quality assessment of this herb. This developed method by having a combination of chromatographic ngerprint and quantication analysis could be applied to the quality control of Rhizoma Belamcandae.
Synthesis of mangiferin
V.K. Bhatia, T.R. Seshadri Department of Chemistry, University of Delhi, Delhi, India
Mangiferin was synthesised by nuclear glucosylation somewhat analogous to nuclear methylation. It was achieved by the reaction of 1,3,6,7-tetrahydroxyxanthone with tetra-O-acetyl--D-glucopyranosyl bromide. The former was obtained by condensing 2,5-dihydroxy-4-methoxy benzoic acid with anhydrous phloroglucinol in the presence of zinc chloride and phosphorcus oxychloride followed by demethylation using hydriodic acid.This synthesis confirms that the glucoside has the -configuration.
175
Study On Mango Leaf and Mangiferin Rapid Identification of Polyphenol C-Glycosides from Swertia franchetiana by HPLC-ESI-MS-MS
Yanguo Sun, Xi Zhang, Xingya Xue, Yan Zhang, Hongbin Xiao, Xinmiao Liang* Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, P.R.China
High-performance liquid chromatography coupled to positive ion electrospray ionization tandem mass spectrometry (MS) and diode array detection was employed to identify the polyphenol C-glycosides in the extract of Swertia franchetiana, a traditional Chinese/Tibetan herb. The neutral loss scan of the extract of S.franchetiana using the characteristic losses of 120 and 150u provided a detailed profile of the polyphenol C-glycosides in the complex mixture. Online UV spectroscopy along with MS-MS and MS-MS-MS mass spectra analysis produced with and without insource collision induced dissociation was contributed to discriminate and identify the polyphenol C-glycosides. Three xanthone C-glycosides (i.e., mangiferin, isomangiferin, and 1, 6, 7-trihydroxyl-2-C-glucosexanthone) and three flavone C- glycosides (i.e., isoorientin, isovitexin, and swertisin) were tentatively identified. Isomangiferin and 1, 6, 7-trihydroxyl-2C-glucose-xanth-one were for the first time found in this plant.
Secoiridoids and Xanthones from Gentianella nitida

Marie-Aleth Lacaille-Dubois*1, Katja GalIe2, Hildebert Wagner2
1 2
Laboratoire de Pharmacognosie, Facult de Pharmacie, Universit de Bourgogne, Dijon Cedex, France Institut fr Pharmazeutische Biologie der Universitt Mnchen, Mnchen, Germany
From Gentianella nitida twelve known metabolites were isolated and identified by HPLC-(UV-and/or by spectroscopic methods as secologanoside, amaroswerin, amarogentin (secoiridoids), isoorientin (C-glucosylflavone), mangiferin, demethylbellidifolin 8-O-glucosicle, norswertianine 1-O-glucoside. swertianine 1-O-primeveroside, swertianine 8-O-glucoside, norswertianine, demethylbellidifolin,and swertianine (xanthone glycosides and aglycones). Secologanoside is reported here for the first time in Gentianaceae species; the antioxidant mangiferin was obtained as the major compound in good yield.
176
Study On Mango Leaf and Mangiferin Simultaneous determination of bioactive xanthone glycosides and
norlignans from ethanolic extract of Anemarrhena asphodeloides by liquid chromatography

M. Nurul Islam1, Hye Hyun Yoo1, Jun Lee2, Joo Won Nam2, Eun Kyoung Seo2, Changbae Jin1, Dong-Hyun Kim1
1 2
Korea Institute of Science and Technology, Doping Control Center, Seoul, Korea; Ewha Womans University, College of Pharmacy and The Center for Cell Signaling and Drug Discovery
Research, Seoul, Korea
The rhizomes of Anemarrhena asphodeloides Bunge (Liliaceae) are prescribed as crude drugs in herbal medication for the treatment of various diseases such as diabetes, inflammation, and platelet aggregation inhibition. A simple, sensitive, and precise reversed-phase liquid chromatographic method was developed to study the quantitative determination of 5 bioactive compounds from these rhizomes, namely, neomangiferin, mangiferin, isomangiferin, nyasol, and methylnyasol. Chromatographic analysis was performed on Capcell Pak C18 column (1504.6 mm, 3m) with a mobile phase consisting of acetonitrile, methanol, and 0.1% formic acid at a flow rate of 1.00 mL/min. Quantitation was performed using a UV-visible detector at 260 nm. The method for the determination of reported medicinal agents was accurate and reproducible. Excellent linear behavior was observed over the investigated concentration range of 2.5-100.0 g/mL for neomangiferin; 1.5-60.0 g/mL for mangiferin; 0.5-20.0 g/mL for nyasol; and 0.2-20.0 g/mL for methylnyasol; correlation coefficient >0.99. The intraday and interday precision over the concentration range of compounds was <6.6% (relative standard deviation) and accuracy was between 94.9 and 109.3%. This method can be successfully applied for the analysis of medicinal compounds from the ethanolic extract of A. asphodeloides Bunge.
177
Study On Mango Leaf and Mangiferin Simultaneous determination of phenols in Radix Polygalae by high performance liquid chromatography: quality assurance of herbs from different regions and seasons
Jun Li1Xiaobing Dong1Yong Jiang2Qiutao Gao1Zhiyong Jiang1AnnaW. H. Cheung1Ran Duan1Tina T. X. Dong1Pengfei Tu2KarlW. K. Tsim1
1
Department of Biology and Center for Chinese Medicine, The Hong Kong University of Science and
Technology, Hong Kong

2
Department of Natural Medicines, School of Pharmaceutical Sciences, Peking University Health Science
Center, China
Radix Polygalae, roots of Polygala tenuifolia or of Polygala sibirica, is a Chinese herbal medicine commonly used to prevent dementia. Reliable chemical markers for quality assurance of this herb are missing. Here, a high performance liquid chromatography method coupled with diode array detection was developed to simultaneously determine nine different phenols in Radix Polygalae, including sibiricose A5, sibiricose A6, glomeratose A, tenuifoliside A, glomeratose D, 3',6-di-O-sinapoyl sucrose ester, mangiferin, polygalaxanthone III, and polygalaxanthone XI. By using two different detection wavelengths in the HPLC analysis, the developed method was able to determine the phenols with excellent resolution, precision, and recovery. This established method was therefore applied to determine the amounts of phenols in thirty-two samples from different cultivation regions and harvest seasons in China, and significant variations were revealed. The amounts of phenols in the roots of P. tenuifolia collected in Shanxi and Shannxi Provinces were markedly higher than in roots collected from other Provinces. Moreover, the samples harvested in the spring contained higher contents of phenols than those collected in other seasons.
178
Study On Mango Leaf and Mangiferin Simultaneous estimation of mangiferin and four secoiridoid glycosides in rat plasma using liquid chromatography tandem mass spectrometry and its application to pharmacokinetic study of herbal preparation
Satyendra Suryawanshi1 , R.K. Asthana2, R.C. Gupta 1
1 2
Extracts from Swertia chirata (family Gentianaceae) have antidiabetics and antioxidant activity, largely attributed to the avonoids and secoiridoids, which are a major class of functional components in methanolic extracts from aerial part of plants. In order to facilitate analysis of systemic exposure to S. chirata derived products in animals, we developed a liquid chromatography tandem mass spectrometry (LCMS/MS) based method that is capable of routinelymonitoring plasma levels of avonoids and secoiridoids.An LCMS/MS-basedmethod has been developed for the simultaneous estimation of two bioactive markers, mangiferin and amarogentin along with three other components, amaroswerin, sweroside and swertiamarin in rat plasma. All the analytes including the internal standard (kutkoside) were chromatographed on RP-18 column (250mm4mm i.d.,5m.) coupledwith guard column using acetonitrile: 0.5mMammoniumacetate buffer, pH3.0 asmobile phase at a owrate of 1ml/min in gradient mode. The nal ow to source was splitted in 1:1 ratio. The detection of the analytes was performed onAPI 4000 LCMS/MS system in the multiple reaction-monitoring (MRM)mode. The quantitation for analytes other than the pure markers was based on relative concentration. Themethodwas validated in terms of establishing linearity, specicity, sensitivity, recovery, accuracy and precision (Intra- and Inter-day), freeze-thaw stability, peltier stability, dry residue stability and long-term stability. The recoveries from spiked control samples were >90% for all analytes and internal standard except mangiferin where recovery was >60%. Intra- and inter-day accuracy and precision of the validated method were within the acceptable limits of <15% at low and <10%at other concentrations. The quantitation method was successfully applied to generate pharmacokinetic (PK) prole of markers as well as to detect other components in plasma after intravenous dose administration of herbal preparation in male Sprague-Dawley (SD) rats.
179
Study On Mango Leaf and Mangiferin Structures of New Friedelane-Type Triterpenes and Eudesmane-Type Sesquiterpene and Aldose Reductase Inhibitors from Salacia chinensis
Toshio Morikawa, Akinobu Kishi, Yutana Pongpiriyadacha, Hisashi Matsuda, and Masayuki Yoshikawa* Kyoto Pharmaceutical University, Misasagi, Yamashina-ku, Kyoto, Japan * To whom correspondence should be addressed.
Three new friedelane-type triterpenes named salasones A(1), B(2), and C(3), a new norfriedelane-type triterpene, salaquinone A(4), and a new acylated eudesmane-type sesquiterpene, salasol A (5), were isolated from the 80% aqueous methanolic extract of the stems of Salacia chinensis collected in Thailand. Their stereostructures were elucidated on the basis of chemical and physicochemical evidence. In addition, six constituents, 3,22-dihydroxyolean-12-en-29-oic acid, tingenone, tingenine B, regeol A, triptocalline A, and mangiferin, were found to show an inhibitory effect on rat lens aldose reductase.
Studies on the constituents from the fruits of Phaleria macrocarpa

Shiori Oshimi1, Kazumasa Zaima1, Yosuke Matsuno1, Yusuke Hirasawa1, Toru Iizuka1, Herra Studiawan2, Gunawan Indrayanto2, Noor Cholies Zaini2, Hiroshi Morita1
1 2
Faculty of Pharmaceutical Sciences, Hoshi University, Shinagawa, Tokyo 142-8501, Japan Faculty of Pharmacy, Airlangga University, Jalan Dharmawangsa Dalam, Surabaya, Indonesia
From the fruits of Phaleria macrocarpa, icariside C3 (1), phalerin (2), and mangiferin (3) were isolated and their structures were identied on the basis of spectroscopic data. Icariside C3 (1) showed a slow vasorelaxant activity against noradrenaline-induced contraction of isolated rat aorta. The structure of phalerin (2) was revised as 2, 4', 6-trihydroxy-4-methoxybenzophenone-2- O--D-glucoside.
180
Study On Mango Leaf and Mangiferin Synthesis of mangiferin derivates and study their potent PTP1B inhibitory activity
Hong Gang Hu1,2 , Ming Juan Wang2 , Qing Jie Zhao 2, Shi Chong Yu2, Chao Mei Liu 2, Qiu Ye Wu 2,*
1 2
Department of Chemistry, College of Science, Wayne State University, Detroit, United States Department of Organic Chemistry, School of Pharmacy, Second Military Medical University, Shanghai,
China
Protein tyrosine phosphatase 1B (PTP1B) has received considerable attention from the drug industry as a potential treatment for diabetes mellitus. Mangiferin has been reported to possess signicant antidiabetic activity. Based on the previous study, eight new mangiferin derivates were synthesized and evaluated for their PTP1B inhibitory activity. Some of them displayed good inhibitory activity on PTP1B.
The Major Phenolic Compounds in the Leaves of Cyclopia Species (Honeybush Tea)
Anna Maria De Nysschen1,Ben-erik Van Wyk1 2,Fanie R.Van Heerdens2,Anne Lise Schutte1
1 2
Department of Botany, Rand Afrikaans University,Johannesburg South Africa Department of Chemistry and Biochemistry, Rand Afrikaans University,Johannesburg South Africa
The phenolic compounds of the leaves of Cyclopia species (tribe Podalyrieae) are of both chemotaxonomic and commercial interest,as the leaves are used to brew a herbal drink known as honey bush tea. Despite the commercial importance of Cyclopia, virtually nothing leaves prior to the present work.Methanolic extracts from was known about the chemistry of the
leaves of 22 species were screened for the
presence and distribution of phenolic compounds.Three major constituents of the leaves were identified as mangiferin (a xanthone) and glycosides of the flavanones hesperitin and isosakuranetin.The combination of these three compounds is a unique character for Cyclopia,as none of them are present in any of the other genera of the tribes Podalyrieae and Liparieae. Various combinations of the three compounds occur in the different infrageneric groups, but the species are remarkably similar. These results are thus of chemotaxonomic significance at the generic rather than infrageneric level.
181
Study On Mango Leaf and Mangiferin Temperature and solvent dependent NMR studies on mangiferin and complete NMR spectral assignments of its acyl and methyl derivatives
Shaheen Faizi, Sadia Zikr-ur-Rehman, Muhammad Ali and Aneela Naz International Centre for Chemical Sciences, H.E.J. Research Institute of Chemistry, University of Karachi, Karachi, Pakistan
By employing concerted 1 and 2D NMR techniques, exact NMR spectral assignments have been made of the acyl (2-7) and methyl (8 and 9) derivatives of mangiferin (1) isolated from the leaves of Bombax ceiba. Derivatives 2, 8 and 9 have been reported in literature, while 3-7 represent new compounds. The acetates 2 and 3 were found to be unstable and were converted into the same penta-acetate 4 at room temperature. Extensive NMR studies on mangiferin (1) and its derivatives showed that H-4 exchanges with deuterium of the solvent molecule more easily. This exchange under acidic conditions occurred at that position (C-4) where electrophilic substitution reactions can easily take place. This is the rst report describing the exchange of C-4 proton of mangiferin (1), or any other xanthone, with deuterium of solvent molecules.
Use of NIRS for quantification of mangiferin and hesperidin contents of dried green honey bush (Cyclopia genistoides) plant material
Elizabeth Joubert, Marena Manley, Mariza Botha Department of Food Science, Stellenbosch University, Matieland (Stellenbosch), South Africa
Cyclopia genistoides,normally used for the preparation of an herbal tea,honeybush,is a good source of the bioactive compounds mangiferin and hesperidin and is in demand for the preparation of xanthone-enriched extracts.Near-infrared spectroscopy(NIRS)was used to develop calibration models to predict the mangiferin and hesperidin contents of the dried green plant material.NIRS measurements of plant material and pure compounds were performed in diffuse reflectance mode.The calibration sets for mangiferin and hesperidin contents ranged from 0.7 to 7.21 and 0.64-4.80g/100g,respectively.Using independent validation,it was shown that the NIRS calibration models for the prediction of mangiferin(SEP=0.46g/100g,R2=0.74,and RPD=1.96) and hesperidin(SEP=0.38g/100g,R2=0.72,and
RDP=1.90)contents of the dried plant material are adequate for screening purposes,based on RPD values.
182
Study On Mango Leaf and Mangiferin Using LC/MS/MS to determine matrine, oxymatrine, ferulic acid, mangiferin, and glycyrrhizin in the Chinese medicinal preparations Shiau-feng-saan and Dang-guei-nian-tong-tang
TingtingJong1, Mawrong Lee1,Yicheng Chiang1,Shutuan Chiang2
1 2
Department of Chemistry, National ChungHsin University,Tai chung,Taiwan Chuang Song Zong Pharmaceutical Co.,Ltd.,Li gang Shiang,Pingtung,Taiwan
We have developed a simple, rapid, selective,and reproducible method for the quality control of traditional Chinese medicinal preparations.In this study,we used LC/MS/MS to simultaneously identify and quantify five marker compounds-matrine, oxymatrine, ferulic acid, mangiferin, and glycyrrhizin-in preparations of Shiau-feng-saan and Dang-guei-nian-tong- tang.The calibration curves for the five marker compounds were linear over the concentration range 50-2500ng/mL(R>0.9971).The matrix effect was minimized and the recoveries of the five marker compounds were>90% at a concentration of 1g/mL. Our
experimental data reveal that significant differences exist between samples obtained from different sources.
Variation of active constituents of an important Tibet folk medicine Swertia mussotii Franch. (Gentianaceae) between artificially Cultivated and naturally distributed
HuilingYang,Chenxu Ding,Yuanwen Duan,Jianquan Liu. Qinghai-tibet Plateau Biological Evolution and Adaptation Laboratory,Northwest Plateau Institute of Biology,Chinese Academy of Sciences,Xining,Qinghai,China
Concentrations of seven phytochemical constituents (swertiamarin, mangiferin, swertisin, oleanolic acid,1,5,8-trihydroxy-3-methoxyxanthone,1,8-dihydroxy-3,7-dimethoxyxanthone and 1,8-dihydroxy-3,5dimethoxyxanthone)ofZangYinChen
Swertiamussotii,a
herb used in Tibetan folk medicine)were
determined and compared in plants collected from naturally distributed high-altitude populations and counterparts that had been artificially cultivated at low altitudes.Levels of mangiferin,the most abundant active compound in this herb,were significantly lower in cultivated samples and showed a negative correlation with altitude.The other constituents were neither positively nor negatively correlated with cultivation at low altitude.Concentrations of all of the constituents varied substantially with growth stage and were highest at the bud stage in the cultivars,but there were no distinct differences between flowering and fruiting stages in this respect.
183
Study On Mango Leaf and Mangiferin UV/vis, 1H and 13CNMR spectroscopic studies to determine mangiferin pKa values
Berenice Gmez-Zaleta1, Mara Teresa Ramrez-Silva1, Atilano Gutirrez1,Enrique
Gonzlez-Vergara2, Marisol Guizado-Rodrguez3,Alberto Rojas-Hernndez1

1
Universidad Autnoma Metropolitana-Iztapalapa, Depto.Qumica,rea de Qumca Analtica,
Apdo.Mxico DF, Mxico

2 3
Centro de Qumica,Instituto de Ciencias,Benemrita Universidad Autonoma de Puebla,Mxico Depto.de Qumica Inorgnica,Instituto de Qumica,UNAM,Ciudad Universitaria,Mxico DF,Mxico
The acid constants of mangiferin(anaturalxanthonoid) inaqueous solution were determined through an UV/vis spectroscopic study employing the SQUAD program as a computational tool.A NMR study complements the pKa values assignment and evidences a H-bridge presence on 1-C.The chemical model used was consistent with the experimental data obtained.The pKa values determined with this procedure were as follows:H 4 (MGF)=H 3 (MGF) - +H + ,pK a 1 (6-H)=6.520.06;H 3 (MGF)=H 2 (MGF) 2 - +H+, pK a2 (3-H)=7.970.06;H 2 (MGF) 2- =H(MGF) 3- +H + ,pK(7-H)=9.440.04;H(MGF) 3- =(MGF) 4- +H + , pKa4(1-H)=12.100.01;where it has been considered mangiferin C19H18O11 as H (MGF).Mangiferin UV/vis spectral behavior,stability study inaqueous solution as well as NMR spectroscopy studies:onedimensional1H,13C,2D correlated1H/C performedby(g)-HSQC and (g)-HMBC methods;are also
presented.pKa values determination of (MGF)in aqueous solution is anecessary contribution to subsequent pharmacokinetic study,and a step towards the understanding of its biological effects.
Quality standard research on Mangiferin crude drug

Jiagang Deng, Yong Chen*, Qin Wang, Linan Pang, Li-Li, Bing Li, Yanling Li Faculty of Pharmacy, Guangxi Traditional Chinese Medical Univerisity. Nanning, P.R.China
In order to establish the quality standard for the mangiferin crude drug from the extract of Mangifera indica L leaf, A TLC method was performed for Identification with Mangiferin as control article, and A HPLC method was performed for the quantitative assay. It showed the same color dots as well as the same Rf value comparing the control article in TLC. In the HPLC quantitative assay, The linear range was 0.195 0.975g ( r = 0.9999), the average recovery was 99.7%, RSD=0.12. The established analysis method was simple, feasible with good reproducibility and could be used for quality control on the mangiferin crude drug from the extract of M angifera indica L leaf.
184
Xanthones from Swertia punctata

Heboja qenkovi1, Katarina avikin-Fodulovi1, VanjaBulatovi1, Ivana Aljani4, Nenad Jurani3, Slobodan Macura3, VlatkaVajs2, Slobodan Milosavljevi4
1 2 3 4
Institute for Medicinal Plant Research, Belgrade, Yugoslavia Institute for Chemistry, Technology and Metallurgy, Belgrade, Yugoslavia Department of Biochemistry and Molecular Biology, Mayo Foundation, Rochester, USA Faculty of Chemistry, University of Belgrade, Belgrade, Yugoslavia
Isolation of 1-O-primeverosyl-3,8-dihydroxy-5-methoxyxanthone and 1-O-gentiobiosy-l-3,7-dimethoxy-8-hydroxyxanthon,along with five known xanthones, isobellidifolin, methylbellidifolin,isoswertianin,methylswertianin and norswertianin-1-O--D-glucoside,from the roots of Swertia punctata is reported.In the aerial parts four xanthones, bellidifolin, methylbellidifolin, swertianolin and Mangiferin, and favone-C-glucoside, isoorientin were identified, The chemotaxonomic and harmacological significance of these results is discussed.
Study on the extracting method of mangiferin in Anemarrhena asphodeloides Bge and Comparison of content of mangiferin in hair, skin and meat of Anemarrhena asphodeloides Bge
De Ji, Tulin Lu, Seng Song Nanjing University of Traditional Chinese Medicine, Nanjing, P. R. China
A HPLC method has developed to study the contents of mangiferin exacted by different extracting method and different extraction solvents, and then compare the contents of mangiferin in hair, skin and meat of Anemarrhena asphodeloides Bge. The results shown using ultrasound extraction method with 50% alcohol as extraction solvent can entirely extract mangiferin in hair, skin and meat of Anemarrhena asphodeloides Bge, and the extraction method is convenient and easy to use. The content of mangiferin in hair, skin and meat of Anemarrhena asphodeloides Bge is 0.298%, 0.538% and 0.826% respectively. This method is simple, rapid, accurate and can be repeated.
185
Study On Mango Leaf and Mangiferin Study of extraction and stability of yellow pigment from mango leaves
Guofeng Wei, Jinsheng Cheng, Zuliang Huang, Youcheng He Department of Applied Chemistry, Youjiang Medical College for Nationalities, Baise, P. R. China
The stability of the extracted pigment was also analyzed Results indicated that the best extraction solvent is 60 ethanol-water solution the ratio of raw material to solvent is 1:9 (g:mL) the extraction temperature is 60, and 1.5 hours of refluxing time for twice extraction. The extraction rate of the pigment is 81.23. The yellow pigment of mango leaves was stable when treated with acid and heatNa Zn oxidant and food additives (such as glucosecitric acid and sodium benzoate) can not affect the yellow pigment, Fe , Cu and vitamin C affects slightly on the extracted pigment
leaves by performing orthogonal test and using mango leaves as raw materials
+ 2+ 3+ 2+
An optimizedly experimental design was developed for the extraction of yellow pigment from mango
Extraction and identification of total flavone from mango leaves

Yulian Tang1, Haini Li1, Haihua Liu2, Suoyi Huang3, Guofeng Wei3
1
Undergraduate of Grade 2003, Department of Clinical Laboratory Medicine, Youjiang Medical College
for Nationalities, Baise, P. R. China

2
Undergraduate of Grade 2004, Department of Clinical Laboratory Medicine, Youjiang Medical College
for Nationalities, Baise, P. R. China

3
Department of Chemistry, Youjiang Medical College for Nationalities, Baise, P. R. China
In order to make full use of the mango plant resources, a pure physical technological process was used to extract flavones from mango leaves, and the contents of the total flavone were determined by spectrophotometry measurement. The recovery rate of this method was 97.33%. This extraction method is a pure physical process without any pollution, which can be an effective way for extracting flavones from mango leaves.
186
Study On Mango Leaf and Mangiferin Extraction and identification of mangiferin from Mangifera indica leaves
Huayi Huang1, Chaozan Nong1, Lingxiao Guo1, Gang Meng1, Xiliang Zha2
1 2
Guangxi Nationalities Hospital, Nanning, P.R.China Fudan University, Shanghai, P.R.China
Mangiferin was extracted and purificated from Mangifera indica leaves by alcohol percolation and other methods, the extractive mangiferin was identified by polyamide thin layer chromatography, spectral absorption scan analysis, infrared absorption spectrum analysis , high performance liquid chromatography and high performance capillary electrophoresis methods. The extractive mangiferin was golden yellow and showed as needle style crystal under light microscopy examination. Its melting point was 271-273
. Ferric
trichloride and strontium chloride reactions were positive. The Rf value of polyamide thin layer chromatography was 0. 46. Spectrum absorption, infrared spectrum, high performance liquid chromatography, high performance capillary electrophoresis results and all parameters above were similar to that of the control mangiferin. The purity of mangiferin extracted from domestic Mangifera indica leaves is similar to that of the standard confirmed by above determined methods. It can be used in pharmacology research.
Extraction of total flavanone from mango leaves by ultrasonic wave

Suoyi Huang1, Haini Li2, Yulian Tang2, Jingxuan Zhang2
1 2
Department of Chemistry, Youjiang Medical College for Nationalities, Baise, P. R. China Department of Laboratory Medicine, Youjiang Medical College for Nationalities, Baise, P. R. China
An extract method for total flavanone from mango leaves was studied in order to make use of the resources of mango leaves. The flavanones were extracted by ethanol from mango leaves with ultrasonic wave and the extracted flavanone was determined by spectrophotometer. The density content of the total flavanone extracted from mango leaves was 2900mg/mL and the recovery rate was 100. 2%. This method is a purely physical process which has not any pollution and can be an ideal way to extract the flavanone from mango leaves.
187
Study On Mango Leaf and Mangiferin Optimized procedures for quercetin extraction from Mangifera indica Linn. leaf based on orthogonal design
Hang Dai1,Xiaotao Hou1,Lixia Zhou1,Yinfang Shen2
1 2
Guangxi Traditional Chinese Medical University,Nanning,P.R.China Yunshang Science and Technology Research Institute Company Limited,Nanning,P.R.China
In order to optimize the procedures for extracting quercetin from Mangifera indica Linn. leaf, the extraction procedures were evaluated by quercetin yield determined by HPLC based on the orthogonal test. The main factor influencing quercetin extraction was the contents of sulfuric acid.The optimal condition for quercetin extraction was the addition of 40-time volume of 5% sulfuric acid with 60-minute ebullition hydrolization.This method is simple and stable.
Determination of mangiferin and homomangiferin in almond leaves by HPLC

Shengbo Wang, Xu Feng, Chengxia Xu, Lili Wang, Jieping Qin Guangxi Traditional Chinese Medical University, Nanning, P.R. China
A HPLC method for the simultaneous determination of mangiferin and homomangiferin in almond leaves has been developed and four samples of almond leaves from different areas have been determined. The separation was performed on a Elite Hypersil C18 column (5m, 4.6mmIDX250mm), with a gradient system of acetonitrile-0.1%H3PO4 as mobile phase at a flow rate of 1.0 mL/min. The detective wavelength was 258nm, the column temperature was 30
.The linear ranges of mangiferin and homomangiferin were
good within the range of 0.0254-0.508ug/uL(r=0.9999) for mangiferin and 0.000960-0.0192ug/uL (r=0.9999) for homomangiferin. The average recovery rate was 100.7% (RSD=2.8%) and 100.6% (RSD=2.6%) respectively. The method is simple and accurate with good reproducibility. It can be used to determine the contents of mangiferin and homomangiferin in almond leaves.
188
Study On Mango Leaf and Mangiferin Determination of gallic acid in mango leaves by HPLC
Xu Feng, Shengbo Wang, Jiagang Deng, Jieping Qin Guangxi Traditional Chinese Medical University, Nanning, P.R. China
A method for determining the contents of gallic acid in mango leaves from different producing areas and breeds was established. The separation was performed on Agilent Eclipse XDB-C18(4.6mm150mm, 5m) column with methanol 0.1%H3PO4 (0.1%triethylamine) (5 1.0mL/min.The detection wavelength was 270nm.
95) as mobile phase at flow rate of The liner range of gallic acid was 0.00512g/L
0.06144g/L(r=0.999 7), the average recovery rate was 98.3%and RSD was1.7%(n=6). The contents of gallic acid in mango leaves from different producing areas and breeds were different. The method is sensitive, accurate, quick, characteristic and reproducible.
Study on the content of mangiferin in mangifera indica L. from different areas

Jiagang DengXu Feng, Qin Wang, Jie ping Qin, Yong Ye Guangxi Traditional Chinese Medical University, Nanning, P.R. China
A method was established to determine Mangiferin in the leaves of Mangifera indica L. from different areas. Mangiferin was separated by Luna 5um C18 (2) column (4.6mmID250mm) with a mobile phase of methanol- 0.1% phosphoric acid solution (30:70) and detected at a wavelength of 258nm. The calibration curve was linear in the range of 0.132 2.63g (r=0.99999). The average recovery rate was 98.7% and RSD=2.6% (n=6). The content of Mangiferin in sample of Guire No.82 from Nanning was the
highest among the fourteen samples. The content of Mangiferin in the leaf of Mangifera indica L. was very different due to the different sample-variety and sample-region.
189
Study On Mango Leaf and Mangiferin Determination of mangiferin and homomangiferin in manggo leaves by HPLC
Xu Feng, Shengbo WangJiagang DengJieping Qin
Guangxi Traditional Chinese Medical University,Nanning, P.R. China
A HPLC method for the determination of mangiferin and homomangiferin in manggo leaves was establishe. The separation was performed on Elite Hypersil ODS column (4.6mm250mm, 5m); mobile phase: acetonitrile-0.1%H3PO4 in a gradient mode; flow rate: 1.0mL/min; detective wavelength: 258nm; column temperature: 30 homomangiferin 0.000960
0.0192g/l
, sample volume: 5L. There was good leaner relationship of mangiferin and within the range of mangiferin: 0.0254 0.508g/l (r=0.9999), homomangiferin
(r=0.9999). The average recovery rate were 101.7% (RSD=2.0%), 101.0%
(RSD=1.7%). It preliminarily established a method for the determination of mangiferin and homomangiferin in manggo leaves by HPLC. It can further improve the quality control system of manggo leaves.
Determination of impurity of homomangiferin in raw medicine of mangiferin by HPLC

Xu Feng, Jiagang Deng, Jieping Qin, Jiaxi Xi , Weidong Zhong, Shengbo Wang Guangxi Traditional Chinese Medical University, Nanning, P.R. China
A HPLC was used to determine Homomangiferin in raw medicine of mangiferin. homomangiferin
68)and detected at a wavelength of 258 nm. The calibration curve was linear in the range of 0.042 40.848g(r=0.999 9). The average recovery rate was 97.8%and
methanol-0.1%phosphoric acid solution(32 RSD=2.2% (n=6). The method is simple, sensitive and precise.It can be used for the determ ination of homomangiferin in raw medicine of Mangifierin..
was separated on a Hypersil ODS 5m column(4.6 mm ID250 mm) with a mobile phase of
190
Study On Mango Leaf and Mangiferin Comparison of HPLC fingerprint among different tissues of Mangifera indica L.
Xu Feng, Jiagang Deng, Jieping Qin, Xinhua Wang, Wei Zhang Guangxi Traditional Chinese Medical University, Nanning, P.R. China
To study HPLC fingerprint of methanol extracts from different tissues of Mangifera indica L. in various species and make a comparison among them. A waters symmetry C18 (5m, 4.6mm250 mm) column was applied with 0.1% phosphoric acid as mobile phase in a gradient mode.The flow rate was 1.0mL/min, the temperature of column was 25
, and the wavelength of detection was 216nm. The HPLC
fingerprint of Mangifera indicaL. in different tissues showed great difference. The Homogeneity should be taken into consideration when study the fingerprint of caulis herbs.
Comparison research on the content of mangiferin between manggo leaf and manggo branch
Jiagang Deng, XuFeng, Qin Wang, Jieping Qin, Yong Ye, Feng Chen Guangxi Traditional Chinese Medical University,Nanning, P.R. China
The contents of mangiferin in the leaves and branches of mango tree from different collecting area and different varieties were separated by a Luna C18(2)(5m,4.6 mm ID250mm) column with a mobile phase of methanol-0.1%phosphoric acid solution (32 68) and detected at a wavelength of 258 nm. There was good linearity relationship between the samples and peak area of mangiferin within the range of 0.132-2.630g (r=0.99999), the average recovery rate was 99.36%, RSD1.3% (n=6). There are great difference of mangiferin between the manggoes in leaves and branches from different producing area and breeds.
191
Study On Mango Leaf and Mangiferin Determination of gallic acid in the leaves of 4 genera of Mangifera indica L. by RP-HPLC
Xiaotao Hou, Hang Dai, Lixia Zhou Guangxi Traditional Chinese Medical University,Nanning, GuangXi, China
A HPLC method was applied to measure the contents of gallic acid in the leaves of Mangifera indica L. The contents of gallic acid were different in various Mangifera indica L. The contents of gallic acid in the leaves of Tianyangxiangmang were the highest. The contents of gallic acid in the leaves of Hongxiangya were the lowest. The method is simple, reliable, accurate and can be used to control the quality of the leaves of Mangifera indica L.
Determination of mangiferin, neomangiferin in Rhizoma anemarrhenae from different producing area

Wansheng Chen, Li Li, Chuanzhuo Qiao, Fubao Dai College of Pharmaceutical Science, Second military Medical University, Shanghai, P.R.China
In order to study the effects of growing condition on quality of Rhizoma anemarrhenae, the contents of mangiferin and neomangiferin in Rhizoma anemarrhenae from same growing period, different habitats and different organs were determined by high performance liquid chromatography. The separation was performed on a Lichrosorb Rp-C18 10 m (4. 6 mm250 mm) column with 25% (v/v) acetic acid-methanol (70:30) as the mobile phase at a flow-rate of 0.8 ml/min, with detection at 320nm. The method is rapid, accurate and with good reproducibility. The contents of mangiferin, neomangiferin and the ratio of them were particularly different between different habitats. The content of mangiferin was relatively high in roots and leaves. Root of Rhizoma anemarrhenae can be used as medicine, and leaf is one of the natural resources of mangiferin.
192
Study On Mango Leaf and Mangiferin Comparison of mangiferin content in different cultivars of mango leaves
Xuejian Li1, Changlin Mo2, Jianggang Deng1
1 2
Guangxi Trditional Chinese Medical University,Nanning, Guangxi, P.R.China Guangxi Science and Technology, Nanning,Guangxi,P.R.China
In Order to compare the mangiferin content in different cultivars of mango leaf, the contents of mangiferin in mango leaf were determined by HPLC. Mango leaves were collected from Tianyang Guangxi, 16 cultivars of mango and a local species were determined. The content of mangiferin in Tainongyihao leaf was the highest, in Guiqimang leaf was the lowest. From the determination results of mangiferin content, only 3 cultivars were higher than local species, other 13 cultivars were similar to local species. There was no significant difference of mangiferin content between local species and most cultivars.
Determination of mangiferin of the aerial parts in Gentiana manshurica Kitagawa

Keqin Lin1,Xueying Yu1,Haoyou Wang2,Zhijun Ma1,Hongtao Wang1
1 2
Institute of Natural Resources,Heilongjiang Academy of Science,Harbin,China Harbin Normal University, Harbin,China
TLC was used to identify mangiferin. High-performance liquid chromatography was used to determine the content of mangiferin in the aerial parts of Gentiana manshurica Kitagawa. The separation was performed on a NOVA-PACK column (1503.9 mm) with methanol-water (10:90) as the mobile phase at a flow-rate of 1.0 ml/min, with detection at 207 nm. The content of mangiferin was 140 mg/100 g.
193
Study On Mango Leaf and Mangiferin Determination of mangiferin in dejecta of rabbit by RP-HPLC
Chunhui Zeng1,Xiaojiao Pan1,Ke Yang1,Dezhi Tang2,Jiagang Deng1,Liyong Fan3
1 2 3
Pharmacy College of Guangxi TCM University, Nanning, P. R. China Nanning Institute for Food and Drug Control, Nanning, P. R. China Guangxi Sante Medical and Pharmaceutical Co., Ltd., P. R. China
To investigate the excretion p rocess ofmangiferin after single oral administration in rabbit, RP-HPLC was used to determinate the mangiferin in dejecta of rabbits in 6 days after administration. It is resulted that the prototype of mangifein was detected in the first day and the second day after administration, instead of the third day. The fecal excretion rates were 20.32 %, 4.12%, respectively, and the total fecal excretion rate was 24. 44%. The recovery rate of mangiferin was 99. 87%, RSD was 1. 92%. It is conclused that a sensitive accurate and reproducible method for the concentration measurement of mangiferin in the dejecta of rabbit is developed.
Determination of mangiferin in Qingqiliangying injection by RP-HPLC

Tianshan Wang, Yang Pan, Guoxiang Ma, Lian Chen Nanjing University of Traditional Chinese Medicine, Nanjing, P.R.China
A RP-HPLC method has been established for the determination of mangiferin in Qingqiliangying injection. The mangiferin was separated on a Nova-pak C18 column (150mm 3. 9mm) and detected at 258nm, using methanol-tetrahydrofuran-0. 85 %H3 PO4 aqueous solution (500:80:15) as the mobile phase. The average recovery of mangiferin was 100. 9 %. This method is simple, rapid, and well reproducible, and thus very reliable for the quality control of Qingqiliangying Injection.
194
Study On Mango Leaf and Mangiferin Determination of the contents of mangiferin and berberine hydrochloride in the Zishen Pills by RP-HPLC
Ronghua Dai, Jun Gao,Xi Wang, Kaishun Bi School of Pharmacy,Shenyang Pharmaceutical University, Shenyang,P.R.China
A RP-HPLC method was developed to determine the contents of mangiferin and berberine hydrochloride in the Zishen Pills. The mobile phase (1) was: CH3OH:H2O:HOAc (22:78:0.6 ,V:V), the detection wavelength was 320 nm. The mobile phase (2) was: CH3OH:CH3CN:H2O: (CH3CH2)
3N(10:23:70:02
,V:V) , pH = 4 (H3PO4), the detection wavelength was 340nm. The average recovery of
mangiferin was 97.3 %, RSD = 1.9 %( n =5), the average recovery of berberine hydrochloride was 100.8 %, RSD = 2.8 %( n = 5). The methods were simple and accurate, and can be used for the quality control of the Zishen Pills.
Determination of mangiferin in rhizoma anemarrhenae from different habitats by HPLC-UV

Rongrong Ma1,2,* , Yihong Tang2, Chunhui Ma2, Zhixiong Li2
1
Center of Research and Development on Life Sciences and Environmental Sciences, Harbin University of
Commerce, Harbin, P.R. China

2
Shanghai Institutes of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of
Sciences, Shanghai, P.R. China
A method was established to determine the mangiferin in rhizoma anemarrhenae from different habitats. HPLC method is performed on a DiamonsilR C18 (250mm 4. 6mm, 5m) column. The chromatography condition consisted of CH3CN - H2O (15:85V /V ) ( pH=3 ) with the detection wavelengh at 254 nm. The column temperature is at 20
and the flow rate is 1.0 mL /min. HPLC method is developed
for the determination of mangiferin in rhizome anemarrhenae from different habitats. The sample from Tianzhen owns the highest content of mangiferin. This method is simple, reliable, repeatable, and is suitable for the determination of mangiferin in rhizoma anemarrhenae.
195
Study On Mango Leaf and Mangiferin Determination of mangiferin in rhizoma anemarrhenae from different habitats by HPLC
Zhongqin Shen Kunshan Traditional Chinese Medicine Hospital, Jiangsu Kunshan, P.R. China
High-performance liquid chromatography was employed to determine the content of mangiferin in rhizoma anemarrhenae from different habitats. The chromatography condition consisted of the mobile phase was water-acetonitrile-0.85% phosphonic acid (100:25:5), The flow rate was 1.0mL /min and the column temperature was 30
, with the detection wavelengh was 258 nm. The content of mangiferin in
rhizoma anemarrhenae from Hebei, Neimeng, Shanxi, Shanxi, Anhui were 0.05~1.93%. The method is simple, rapid, accurate
repeatable, The content of mangiferin in rhizoma anemarrhenae from different
habitats was differentiation.
Determination of mangiferin and sarsasapogenin in rhizoma anemarrhenae and stir-baked rhizoma anemarrhenae before sprinking salt solution by HPLC
Xiaoping Dang, Chunqin Mao*, Tulin Lu, Jiajia Xu, Jing wang Nanjing University of Chinese Medicine, Jiangsu Nanjing 210046, PR China
HPLC-UV and HPLC-ELSD were employed to determine the content of mangiferin and sarsasapogenin in rhizoma anemarrhenae and stir-baked rhizoma anemarrhenae before sprinking salt solution. The chromatography condition for determination of mangiferin was performed on a Kromasil column (250 mm 4.6mm, 5m). The mobile phase was water-acetonitrile -0.85% phosphonic acid (100:25:5 ). The flow rate was 0.6mL/min. The column temperature was 30
, and the detection wavelengh
was 258 nm.To determination sarsasapogenin, a Kromasil column (250 mm 4.6mm, 5m) was used, the mobile phase was methanol-water (95:5), the flow rate was 1.0mL /min. The column temperature was 30
and the detector was ELSD. The diift tube temperature was 85 . The content of mangiferin and sarsasapogenin in rhizoma anemarrhenae and stir-baked rhizoma anemarrhenae before sprinking salt solution were both to be improved. The method is suitable for the quality evaluation and quality control of mangiferin and sarsasapogenin in rhizoma anemarrhenae and stir-baked rhizoma anemarrhenae before sprinking salt solution.
196
Study On Mango Leaf and Mangiferin Determination of mangiferin and neomangiferin in rhizoma anemarrhenae and its preparation by HPLC
Hongli Zhai1,2,*, Lianna Sun1,2, Wei Lai1,2, Xiaojing Yu1,2, Wansheng Chen2,3
1 2
Department of Pharmacognosy, School of Pharmacy, Shanghai, P.R. China Modern Research Center for Traditional Chinese Medicine, Second Military Medical University,
Shanghai, P.R. China

3
Department of Pharmacy, Changzheng Hospital, SecondMilitaryMedicalUniversity, Shanghai, P.R. China
A HPLC method was established to determine the content of mangiferin and neomangiferin in rhizoma anemarrhenae and its preparation. The determination was performed on a Diamonsil C18 column (250 mm 4.6mm, 5m, Dkima) ,with the mobile phase composed of acetonitrile -water-formic acid (12:88:1.7). The detection wavelength was 317nm, and the flow rate was 0.8 ml/min. The calibration curves of mangiferin and neomangiferin were linear between 2.45 ~ 49.00 g/mL (r =0. 9993), 0.99~37.60g/mL (r =0. 9991) respectively. This method is simple, accurate, reliable and is suitable for determination of mangiferin and neomangiferin.
Determination of mangiferin and neomangiferin in rhizoma anemarrhenae by HPLC

Yonggang Zhou1,*, Sheng Huang2, Li Gu2, Bin Wang3, Guoqing Zhang3, Ziyang Lou2
1 2 3
The 81th Hospital of Nanjing Military Region, Nanjing, P.R. China The Second Military Medical University, Shanghai, P.R. China The Eastern Institute of Hepatobiliary Surgery, Shanghai, P.R. China
A HPLC method was established to determine the content of mangiferin and neomangiferin in rhizoma anemarrhenae. A Zorbax Eclipse XDB-C18 column (250 mm 4.6mm, 5m) with a grient mobile phase composed of 25 mmol/L dihydrophosphate potassium and acetonitrile were used. The decection wavelength was 257 nm and the flow rate was 1.0mL/min. The calibration curves of mangiferin were linear between 14.2~568.5g/mL (r =0.9999), the neomangiferin were 14.8~590.5g/mL. The within-day precision RSD was both less than 4.5% and the inter-day precision RSD was less than 3.9%. The method is simple, rapid, accurate and is suitable for the determination of mangiferin and neomangiferin.
197
Study On Mango Leaf and Mangiferin Determination of chimonin and forsythiaside in Kangbingdu oral liquid by RP-HPLC
Fangmei Li1,*, Zhaozhan Lin2, Songfeng Zhen2, Chenchen Zhu2
1 2
Nantong Jinghua Pharmaceutical Company Limited, Nantong, P.R. China College of Chinese Materia Medica, Guangzhou University of Chinese Medicine, Guangzhou, P.R. China
A RP-HPLC method was established to determine the contents of chimoni and forsythiaside in Kangbingdu Oral Liquid (KOL) and a new quality standard for KOL was developed. The chromatographic conditions were as follows: Kromasil RP-C18 (250 mm 4.6mm, 5m) column was used, a mixture of acetonitrile - 0.1% acetic acid (15:85, V/V ) served as mobile phase, the flow rate was 0.80mL/min, and detective wavelengths of time process were 025 min:258 nm, and 2530 min:277 nm. Theoretical plate number of chimoni was over 6000 and that of forsythiaside was over 5000.The chimonin showed a good linearity in the range of 48.8ng~1525ng (r = 0.9994), and the linear range of forsythiaside was in the ranged of 56ng~1750ng ( r =0.9995 ). The mean recovery of chimonin and forsythiaside was 97.65% and 99.21% respectively. The content of chimonin in the 10 batches of samples was in the range of 14.40~20.41g/mL, and that of forsythiaside was in the range of 28.70~36.01g/mL. The method has been proved to be simple, stable and reproducible, and can be applied for quality control of the Kangbingdu Oral Liquid.
Determination of Mangiferin in Mango peel by RP-HPLC

Min-qi Huang1, Han-shen Zhen2, Wan-na Xiong2, Jian-ping Jiang2, Mei-qiong Dang1
1 2
Guangxi institute of Health and Cadre Management,Nanning, ,Guangxi,P.R.China Guangxi Traditional Chinese Medical University , Nanning , Guangxi , P.R.China
In order to establish a method for the determination of mangiferin in Mango peel, a HPLC method was established, and a Hanbon Lichyopher C18 (4.6250mm,5m) column was used. The mobile phase was methanol-0.3%H3PO4 (32:68) and the flow rate was1ml/min. The UV detection wavelength was 258nm ,The linear ranges of mangiferin were in the range of 0.4~0.8ug with equation of Y=1.065+2.035X, r=0.9999 (n=5) The average recovery of mangiferin was 97.8 with RSD of 1.85
. This method is simple accurate reproducible. It was found that the Mango peel in Bai-se, Nanning, and Tian-yang county
have the highest content of Mangiferin.
198
Study On Mango Leaf and Mangiferin RP-HPLC determination of mangiferin in the leafs of Folium Mangiferae sampled in different months and regions
Haibin HUANG , Xuejian LI , Qiuyun LING Pharmaceutical factory of Guangxi Traditional Chinese Medical University, Nanning , Guangxi , P.R.China
To develop a new method for determination of Mangiferin in the leaves of Folium Mangiferae. Useing this new method, mangiferin in F. mangiferae sampled in different months and in different regions was determinated. A RP-HPLC method was set up, a Shim pack CLC-ODS column was used, methanol-0. 05 molL
-1
H3 PO4 (65 134, pH 3. 5) as mobile phase, the flow rate was 1 mL min 1, with 258 nm as
detection wave, at room temperature. Samples of F. mangiferae collected in Nanning, Qinzhou and Tianyang, Guangxi province from January to December were determined respectively. The average recovery of the RP-HPLC was 99. 2 %, RSD = 1. 05 %( n = 5). The contents of mangiferin in F. mangiferae was statistically different due to different Sample-regions or sample -time. This RP-HPLC method is simple, specific and exact. The contents of mangiferin in the leaves of F. mangiferae sample in Nanning and Tianyang were statistically similar, but higher than that in Qinzhou. The contents of mangiferin in the leaves of F. mangiferae sampled in July to October were higher than that in the other months. The content in September was the highest, the content in February was the lowest .
Determination of the contents of mangiferin in the roots and stems of Rhizoma Anemarrhenal at different harvest dates
Yongfu Hong etc.
A TLC-UV method was used to determine and compare the contents of mangiferin in the roots and stems of Rhizoma Anemarrhenal at different harvest dates. Mangiferin was separated by TLC and then detected at 258nm to count the contents, the linear ranges was in the range of 1~10ug, y=0.0152x+0.007. The determination results shown the root and stem of Rhizoma Anemarrhenal has the highest content of mangiferin (1.26%) in April, and the lowest mangiferin content was in March (0.12%).This study provided some basis data for making use of Rhizoma Anemarrhenal and researching the metabolic process of mangiferin.
199
Determination of mangiferin and neomangiferin in Rhizoma Anemarhenal using RP-HPLC

Jianjun WANG, Ziyang LOU, Yutian WU Department of Pharmaceutical Analysis, School of Pharmacy, the second Military Medical Unitersity, Shanghai,P.R. China
To establish a HPLC assay for the determination of mangiferin and neomangiferin in Rhizoma Anemarrhenal as well as its total flavones. The determination was performed on a C18-ODS column (200mm4.6mm, 5um), with the mobile phase of acetonitrile -0.05 mol.L-1 NaH2PO4 (adjusting PH to 3.20) (10:90). The detection wavelength was at 317 nm. Both of mangiferin and neomaniferin showed good linearity over the ranges of 2.0-40.5 ug.mL-1 (r=0.9997) and 3.0-59.2 ug.mL-1, (r=0.9997), respeclively. The recoveries were 96.4% (RSD=1.79%, n=5) for mangiferin and 97.1% (RSD=2.14%, n=5) for neomangiferin. This method was easy , rapid , accurate , and sensitive . It will be employed for detemining mangiferin and neomangiferin in herbs and tablets.
Determination of Mangiferin and Polysaccharide in Rhizoma Anemarrhenal from Different Origin

Honghui Guan 1, Qiuping Guo 1 ,YingGao 2, Xiaohua Yao 1
1 2
Pharmacy College,Guang zhou University of TCM,Guang zhou, Guang dong, P.R.China New Drug Research and Development Center,Guang zhou University of TCM,Guang zhou, P.R.China
To establish a method for determing mangiferin and Polysaccharide in Rhizoma Anemarrhenal from Different Origin, we used a HPLC method with Dumas C18 (4.6250mm,5m)column, the mobile phase was methanol-3% acetic (45:55) at a flow rate of 1ml/min. We found that Mangiferin in Rhizoma Anemarrhenal from He-Bei, An-Hui, Guang-Dong was 0.50%~1.18%, The average recoveries of mangiferin was 101.54
1.29~7.74ug (r=0 9995); Polysaccharide in Rhizoma Anemarrhenal from He-Bei,An-Hui,Guang-Dong was 0.52%~1.91%, The average recoveries of mangiferin was 99.15
with RSD of 2.33 (n=6);The linear ranges of mangiferin was in the range of with RSD of 1.94(n=6). So we
made a conclusion that it was different in the contents of Mangiferin and Polysaccharide in Rhizoma Anemarrhenal from Different Origin.
200
Study On Mango Leaf and Mangiferin Quantitative determination of four effective components in Swertia delavayi by HPLC
Conglong Xia, Guangming Liu, Yin Wang Department of Pharmacy,Yunnan Dali University,Dali,P.R.China
A method was established to determine the contents of swertiamarin, gentiopicroside, mangiferin and oleanolic acid in Swertia Delavayi. The chromatographic condition of swertiamarin, gentiopicroside, mangiferin was that the ZORBAX SB-C18 ( 4.6 mm150 mm , 5m) was used, the mobile phase
consisted of CH3OH-H2O( 0.1% H3PO4 ) gradient elution (CH3OH %: 0 min 20 % ,30 min 30 %) , the flow rate was 0.60 mLmin-1, the UV detection wavelength was 240nm , the temperature of column was 30
; .
The chromatographic condition of oleanolic acid was that the ZORBAX SB-C18 (4.6mm 150 mm , 5m) was used , the mobile phase consisted of CH3CN- H2O( 0.1% H3PO4 ) (75 25) , the flow rate was 1.0 mLmin-1, the UV detection wavelength was 210 nm , the temperature of column was 40
Swertiamarin , gentiopicroside , mangiferin , oleanolic acid were in good linearity over the range of 0.155 19g ; 0.018 6 - 1.24g ;0.00680 - 1.36g ; 0.15 - 3.9g.respectively (r= 0. 9999 , 0.9999 , 0.9998 , 0.9996 ) , The average recoveries were in the range of 99 % and 103 % ,with RSD of less than 2%. The
method is simple , sensitive , accurate and reproducible and can be used to control the quality of Swertia Delavayi.
Determination of mangiferin in Liyan tablets by HPLC

Ping Xiao1, Xingzhen Huang2
1 2
The Fourth Affiliated Hospital of Guangxi Medical University, Liuzhou, P.R.China Guangxi Medical University, Nanning, P.R.China
The content of mangiferin in Liyan tablets was determined by RP-HPLC with kromasil - C18 column, the mobile phase consisted of acetonitrile-0.5% phosphoric acid - triethylamine ( 20:79.5:0.5) with a flow rate of 1.0 mLmin-1, and the injection volume was 20L. External standard method was used and the wavelength of the detector was set at 258 nm. The linear range was 5.12 - 51.20 mgL-1, the correlation coefficient was 0. 9999. The average recovery and the relative standard deviation were 102.22 % and 1.10 % respectively. The method is simple and accurate and the precision is good . It is proved to be suitable for the quality control for Liyan tablets.
201
Study On Mango Leaf and Mangiferin Determination of mangiferin in Zhibai Dihuang Pill by HPLC
Yuerong Chen, Haiqing Liu Qinghai Institute for Drug Control, Xining, P.R.China
The content of mangiferin in Zhibai Dihuang Pill was determined by HPLC with Agilent ODS C18 column (4.6 mm 250 mm , 5m), the mobile phase consisted of CH3OH-0.1% phosphoric acid ( 25:75) with a flow rate of 1.0 mLmin-1, and the wavelength of the detector was set at 316 nm. The linear range was 0.132- 0.660 mgL-1 and the correlation coefficent was 0. 9997. The average recovery and the relative standard deviation were 98.14 % and 0.55 % respectively. The method is simple and accurate and the precision is good . It is proved to be suitable for the quality control method for Zhibai Dihuang Pill.
Dertermination content of mangiferin and sarsasapogenine in Rhizoma Anemarrhenae from different areas
Xiuqing Hou1, Chunqin Mao2, Tulin Lu2, Shen Song2
1 2
China Pharmaceuticul University, Nanjing, P.R.China Nanjing University of ChineseMedicine, Nanjing, P.R.China
To establish the determination of mangiferin and sarsasapogenine in Rhizoma Anemarrhenae from different areas. The content of mangiferin and sarsasapogenine in Rhizoma Anemarrhenae were determined by HPLC and HPLC-ELSD. The mobile phase were water-acetonitrile-0.85% phosphoric- acid (100:25:5) and methanol-water (95:5) , with the flow rate were 0.6mL /min and 1.0mL /min, at 30
. The detection
wavelength of mangiferin was at 258 nm. An evaporative light - scattering detector (ELSD) was used as detector to determine sarsasapogenine with the drifttube temperature of 85 . The content of mangiferin and sarsasapogenine in Rhizoma Anemarrhenae from different areas were 0.11% 0.73% and 1.51%
1.73%. The method can be used for the quality control of Rhizoma Anemarrhenae from different areas, the result have provide a scientific basis for processing Rhizoma Anemarrhenae .
202
Study On Mango Leaf and Mangiferin Separation of mangiferin by high performance capillary electrophoresis
Huayi Huang,Chaozan Nong,Lingxiao Guo,etal Department of Central Laboratory and Tumor Molecular Cell Biology Laboratory , Guangxi Nationalities Hospital , Nanning , P.R.China
In order to explore an effective separation of Mangiferin, A free solution capillary zone electrophoresis (CZE) was used to analyze Mangiferin extracted from Mangifera indica leaves. The migration time was similar to that of sigma reagent when monitored at 254 nm, a better separation was obtained using pH 7. 4 boric acid solution-methanol (1:0. 3) as buffer solution. The method is useful for clinical analyses of Mangiferin.
HPLC determination of mangiferin content in commercial Rhizoma Anemarrhenae

Yuerong Chen Qinghai Institute for Drug Control, Xining, China
In order to establish an HPLC method for the determination of mangiferin content in Rhizoma Anemarrhenae, an external method with Elite-ODS column(4.6 mm 150 mm , 5m) and methanol-0.1% phosphoric acid solution(27:73) as mobile phase was adopted. The detective wavelength was 316 nm and the flow rate was 1.0mL /min. The linear range was 0.214~1.07g and the correlation coefficient was 0.9997. The average recovery(n=3) average of low middle and high of concentration 0.86 were
99.75%,98.47%,96.96%,with
relative
standard
deviations
%,0.91%,0.56%
respectively. The method is simple,accurate and the precision is good . It can be used for the quality control of Rhizoma Anemarrhenae.
203
Study On Mango Leaf and Mangiferin Comparison of Mangifern in Contents in Different Parts of Mango Tree
Xuejian Li, Jiagang Deng Guangxi College of TCM, Nanning Guangxi 530001
In order to compare the mangiferin contents in different parts of mango tree, HPLC method was used to determine the mangiferin contents in root, stem, bark, leaf, branch of mango tree. Mangiferin contents in bud, tender leaf, tender branch and mature fruit were similar to that in mature leaf, but those in flower, stem, root, coarse branch were obviously lower and that in green fruit were significantly higher than that in mature leaf. Mangiferin contents in different parts of mango tree are different.
The physiological and biochemical change induced by mangiferin accumulation in Mango tree
D.K.Charkrabarti
Mangiferin is the natural metabolite of Mango. Its accumulation in regeneration buds effects the reproductive growth turning to vegetable growth. The test was performed on the research of the mango physiological change induced by mangiferin. And relationship between normal or disorder metabolism and the mangiferin accumulation in dysplastic type mango tree was confirm in this paper. High concentration mangiferin could lower the activity of peroxydase, catalase, amylase and IAA oxidase but increase the activity of polyphenol oxidas and convertase. Mangiferin accumulateion increased the photosynthesis but lower the transpiration and respiration. Mangiferin could increase the content of chlorophyl, carbohydrate, total nitrogen, protein nitrogen, nucleinic acid(RNA & DNA), heteroauxin.
204
Study On Mango Leaf and Mangiferin Optimization of Extraction Technology for Total Saponin in Mango Leaf
Yanqing Tang1, Danping Qiu1, Yonglin Luo1, Suoyi Huang2
1
Undergraduate of Grade 2006 ,Department of Clinical Laboratory Medicine ,Youjiang Medical College
for Nationalities ,Baise,P.R.China

2
Teaching and Research Section of Chemistry ,Department of Pharmacy ,Youjiang Medical College for
Nationalities ,Baise,P.R.China
The influence factors which affected the extraction technology of total saponin in mango leaf, included alcohol concentration, volume, extracting time and temperature, were studied with alcohol as extraction solvent by ultrasonic wave. The optimized extraction technology for the total saponin of mango leaf was as followed : 30 times of amount of 75 % alcohol, extracted at 70 ultrasonic was 100 MHz.
for 35 min, the power of
Rapid determination of mangiferin and neomangiferin in Anemarrhena asphodeloides Bge. By capillary zone eectrophoresis with UV detection
Jianjin Wang, Shanlei Qiao, Yifeng Chai, Ziyang Luo, Yutian Wu Department of Phamaceutical Analysis, School of Pharmacy , the Second Military University, Shanghai, P.R.China
A rapid capillary zone electrophoresis method was, for the first time, developed for the simultaneous determination of mangiferin and neomangiferin in the Chinese herbal extract from Anemarrhena asphodeloides Bge. Optimun separation was achieved with 30mmoLL-1 Borax at pH 9.18. The applied voltage was 25 kV and the capillary temperature was kept constant at 30
and the detected wavelength
was 214nm. Uncoated fused silica capillary column 50m50cm (effective length 41.5 cm ). Cinnamic acid was used as the internal standard.Regression equation revealed good linear relationship (correlateon coefficient: 0.9995 and 0.9994) between the ratio of peak area of each compound (mangiferin and neomangiferin) and its concentrateion (concentration range: 8.1-162.0, 5.9-118.0 gmL-1). The relative standard deviations of relative peal area were less than 4.1%. The contents of the two flavonoids in Anemarrhena asphodeloides Bge. were successfully determined in 6min, with satisfactory recovery and repeartability.
205
Study On Mango Leaf and Mangiferin Isolation and Identification of Oleanolic acid and Magiferin from Swertia punicea hemsl
Guangming Liu, Yongshou yang, Guangping Dong, Huai Xiao Pharmaceutical Department of Dali Medical college, Dali,P.R.China
Using pressed silica gel cylinder chromatography to extract, isolated and identify the main chemical compounds from Swertia punicea hemsl, a species of Chinese traditional herbs .Using ultraviolet, infared, nuclear-magnetic and mass spectra to analyse and identify the physical and chemical characteristics of the components.13 monomers were isolated from the alcohol extract of Swertia punicea hemsl. Two compounds were identified. Compound 1 was proved to be oleanolic acid and compound 2 was proved to be mangiferin. Other compounds are being identified. Two compounds, oleanolicid and mangiferin were firstly isolated and identified from Swertia punicea hemsl.
Changes of mangiferin and neomangiferin Anemarrhenae before and after processing

Liu Min1,Zhao Baiyun1,Zhao Liang2,Lou Zi yang3,Chai Yifeng1.
1
contents
in
Rhizoma
Department of Pharmaceutical Analysis,School of Pharmacy,Second Military Medical University, Shanghai,P.R.China
Department of Pharmacy,Eastern Hepatobiliary Surgery Hospital,Second Military Medical University, Shanghai,P.R.China
Analysis and Testing Center ,School of Pharmacy,Second Military Medical University,Shanghai, P.R.China
To determine the changes of mangiferin and neomangiferin contents in Rhizoma Anemarrhenae before and after processing. High performance liquid chromatography HPLC was used to determine the changes of mangiferin and neomangiferin contents in Rhizoma Anemarrhenae before and after processing.The mobile phase was ACN: 25mmol/L KH2PO4
pH3.0, gradient elution. The Similarity Evaluation System for Chromatographic Fingerprint of TCM2004 A editionwas used to evaluate the similarity of the 3
regulated and neomangiferin was down regulated afer prosessing.The similarity of the 3 batches was all
batches of Rhizoma Anemarrhenae before and after processing.Mangiferin in the 3 batches was up
decreased(<1).Processing changes the contents of mangiferin and neomangiferin in Rhizoma Anemarrhenae.
206
Dynamic Study of Contents of mangiferin in XiLing Anemarrhena asphodeloides Bge.

Qianliang Chen1,Wenquan Wang1,Changhua Ma1,Jianyong Liu2,Yushan Yin3
1 2 3
BeiJin University of Traditional Chinese Medicine University,BeiJing,P.R.China He Bei Yi County Science & Technology Bureau,Yi County, P.R.China He Bei Yi County TaiHang Plant Products Development Company limited,Yi County, P.R.China
In order to explore the content variation of mangiferin in XiLing Anemarrhena asphodeloides Bge and determine the best collecting time, we carried out a dynamic study of XiLing Anemarrhena asphodeloides Bge which was evaluated by mangiferin yield determined by HPLC. The finding is the change trend of content of mangiferin from March to November. The content change is significant.We found that different collecting time great effected medical quality. This result provides scientific basis for collecting Anemarrhena asphodeloides Bge..
Study on the Factors Affecting the Mangiferin Contents in Mango Leaves

Xuejian Li,Jiagang Deng,Zhenlin Qin Guangxi Traditional Chinese Medical University, Nanning, P.R.China
In order to study the growth factors which affect the mangiferin contents in Mango Leaves, Mango trees were cultivated by controlling growth factors including illumination, water, fertilizer, fructification and graft. Then Mango leaves were collected to determine the mangiferin contents. Graft and fructification did not affect the mangiferin contents in Mango leaves. Less water and/or more illumination increased Mangiferin contents. But the increase in nitrogenous fertilizer reduced mangiferin contents. More illumination and /or low or growth rate increased Mangiferin contents, while graft and fructification made no difference.
207
Structure modification of mangiferin

Hongli Liao1 2,Qiuye Wu2,Honggang Hu2,Zhihe Zang1,Li Song1,Qian Yang1
1 2
School of Pharmacy,Chengdu Medical College,Chengdu,P.R.China School of Pharmacy,Second Military Medical University,Shanghai,P.R.China
In order to search for new antidiabetic compounds,the structure of mangiferin is modified. Using mangiferin as startingmateial,a series of 3,6,7-O-trisubstituted derivatives of mangiferin were synthesized,and then their antidiabetic activities were tested in vitro using mangiferin as the control.A total of 11 target compounds were synthesized,and their structures were all confirmed by H-NMR. Compounds 5 and 11 were shown to have better antidiabetic activities than mangiferin.
Identification and determination of four metabolites of mangiferin in rat urine

Hui Wang, Guan Ye, Chun-Hui Ma, Yi-Hong Tang, Ming-Song Fan, Zhi-Xiong Li, Cheng-Gang Huang* Shanghai Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, PR China
Four metabolites of mangiferin were firstly isolated and identified from rat urine. The structures of the four metabolites were determined to be 1,3,7-trihydroxyxanthone (M-1), 1,3,6,7tetrahydroxyxanthone (M-2), 1,3,6-trihydroxy-7-methoxyxanthone (M-3) and 1,7- dihydroxyxanthone (M-4), respectively. A simple and specific analytical method for determination of the four metabolites in rat urine was developed by high performance liquid chromatography (HPLC). Quercetin was employed as an internal standard. The correlation coefficients of the calibration curves were higher than 0.997, both intra-and inter-day precision of four metabolites were determined and their R.S.D. did not exceed 10%. The accuracy and linear range had been investigated in detail. The cumulative urinary excretions of the four metabolites were measured and the possible metabolic pathway of the metabolites was discussed.
208
Study On Mango Leaf and Mangiferin Stability of mangiferin and factors affecting the stability
Yiping Chen,Xiaojing Niu,Shuangying Chen School of Pharmaceutical Sciences,Guangxi Traditional Chinese Medical University,Nanning,P.R.China
A method was established to determine the stability of mangiferin and analyze its affecting factors in different conditions. The factors affecting the chemical structure of mangiferin in different conditions have been studied by Ultraviolet- visible spectrum under 200-800 nm. Mangiferin was stable when it existed in aqueousphase under slightly acidic and neutral conditions. Furthermore, its aqueousphase at 100
was
unstable. In the processes of extracting and using Mangiferin, metalion should be wiped off, as transitional metalions such as Cu2+
Fe Al
3+
3+
could induce oxidation. Using ultraviolet-visible spectrum we can
evaluate the factors affecting the stability of Mangiferin in different conditions.
Isolation and structure modification of mangiferin from Anemarrhena asphodeloides Bge.

Hongli Liao Second Military Medical University, Shanghai, P.R.China
In order to look for new antidiabetic compounds, we have improved the method of isolating mangiferin and studied the structure modification of mangiferin. With Mangiferin as the lead compound, twenty-one mangiferin derivatives were synthesiezd and their structures were all confirmed by IR, HNMR, NMR and MS. Twenty of them are reported at the first time. Preliminary screening of twenty-one target compounds, most of them can inhibit protein tyrosine phosphatase 1B (PTP1B). Meanwhile, three compounds having high antidiabetic activities were identified. This suggests that mangiferin derivatives perhaps show antidiabetic activities by Inhibiting PTP1B.
209
Study On Mango Leaf and Mangiferin Determination of mangiferin in Zhimu Compounding Granules by HPLC
Ying Peng Department of Pharmacy,The people's hospital of taiping,Dongguan, P.R.China
The content of mangiferin in Zhimu Compounding Granules was determined by HPLC with Diamonsil C18 column (4.6 mm 250 mm , 5m), the mobile phase consisted of acetonitrile -1% CH3COOH ( 20 80, V /V) with a flow rate of 1.0 mLmin-1, the injection volume was 10L and the wavelength of the detector was set at 320 nm. The linear range was 30-150 ng and the correlation coefficent was 0. 9999. The average recovery and the relative standard deviation were 99.1 % and 1.63 % respectively. The method is simple, accurate and the precision is good . It is proved to be suitable for the quality control for Zhimu Compounding Granules.
High-performance liquid chromatographic method for the determination of mangiferin, ikviritin and dihydroquercetin in rat plasma and urine
Sergey V. Geodakyan, Inna V. Voskoboinikova, Jury A. Kolesnik*, Nonna A. Tjukavkina, Litvinenko and Vladimir I. Glyzin Department of Organic Chemistry, Faculty of Pharmaceutical Science, Moscow Medical Academy,, Moscow,Russia Vasiliy I.
The use of reversed-phase high-performance liquid chromatography for the determination of the biologically active plant phenolic compounds mangiferin, likviritin and dihydroquercetin is described. Perchloric acid (35%) was used for deproteinization in the case of mangiferin and methanol for dihydroquercetin. Detection was performed at 254, 275 and likviritin, and acidified 290 nm for mangiferin,
likviritin and dihydroquercetin in plasma, and 365, 312 and 290 nm in urine, respectively. The limit of detection was 0.2 pg/ml for plasma and 0.5 pg/ml for urine.
210
Study On Mango Leaf and Mangiferin Pharmacokinetic study of free mangiferin in rats by microdialysis coupled with microbore high-performance liquid chromatography and tandem masss pectrometry
Ling Lai1,2, Lie-Chwen Lin3,Jer-Huei Lin1, Tung-Hu Tsai2,3*
1 2 3
National Laboratories of Foods and Drugs, Department of Health, Executive Yuan, Taipei, Taiwan Institute of Traditional Medicine, National Yang-Ming University, Taipei, Taiwan National Research Institute of Chinese Medicine, Taipei, Taiwan
Mangiferin (2-b-D-glucopyranosyl-1,3,6,7-tetrahydroxyxanthen-9-one) has been isolated from the herbal root of Anemarrhena asphodeloides Bung showing antioxidative, antiviral, and anticancer effect. An in vivo microdialysis sampling method coupled to microbore high-performance liquid chromatography (HPLC) was employed for continuous monitoring of free mangiferin in rat blood. Microdialysis probes were inserted into the jugular vein/right atrium and brain striatum of Sprague-Dawley rats, and mangiferin at doses of 10,30 or 100 mg/kg were then administered via the femoral vein. Dialysates were collected every 10 min and injected directly into a microbore HPLC system. Mangiferin was separated by a reversed-phase C18 microbore column (1501mm) from dialysate within 10 min. The mobile phase consisted of acetonitrile-0.05% phosphoric acid-tetrahydrofuran (10:75:15, v/v/v) with a flow-rate of 0.05 ml/min. The wavelength of the UV detector was set at 257nm. The limit of quantification for mangiferin was 0.05 mg/ml and in vivo recovery of mangiferin at concentrations of 1, 5 and 10 mg/ml was in range of 37.7-39.8%. The results indicate that the pharmacokinetics of mangiferin at doses of 10-30 mg/kg reveals a linear relation, while doses of 30-100 mg/kg show a nonlinear pharmacokinetic phenomenon. Mangiferin was undetectable in brain dialysate. The proposed method provides a technique for rapid and sensitive analysis of free mangiferin in rat blood and further application in pharmacokinetics tudy. Furthermore, the metabolites of mangiferin in the rat bile were confirmed by LC electrospray ionization (ESI) tandem mass spectrometry (MS-MS).
211
Study On Mango Leaf and Mangiferin High-performance liquid chromatographic method for the determination of mangiferin in rat plasma and urine
Hui Wang1, Guan Ye1, Yi-Hong Tang1, Hai-Yan Zhu1, Rong-Rong Ma2, Zhao-Ling Sun1 and Cheng-Gang Huang1*
1
Shanghai Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, Peoples Republic of China
Center of Research and Development on Life Sciences and Enviromental Sciences, Harbin University of Commerce, Harbin, Peoples Republic of China
A reversed-phase high-performance liquid chromatography assay for mangiferin in rat plasma and urine was developed. Rutin was employed as an internal standard. The mobile phase consisted of acetonitrile-water (16: 84, v/v) containing 3 acetic acid at a flow rate of 1mL/min. Detection was at 257 and 365 nm for mangiferin in plasma and urine, respectively. The limit of quantitation (LOQ) of mangiferin was 0.6 g/mL in plasma, and 0.48 g/mL in urine. The standard curve was linear from 0.6 to 24 g/mL in plasma, and 0.48 to 24 g/mL in urine, both intra- and inter-day precision of the mangiferin were determined and their RSD did not exceed 10 %. The method provides a technique for rapid analysis of mangiferin in rat plasma and urine, which can be used in pharmacokinetic studies.
212
Study On Mango Leaf and Mangiferin Identification of major xanthones and steroidal saponins in rat urine by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry technology following oral administration of Rhizoma Anemarrhenae decoction
Chunhui Ma1, Longxing Wang2, Yihong Tang1, Mingsong Fan1, Hongbin Xiao2* and Chenggang Huang 1*
1
Shanghai Institute of Materia Medica, Shanghai Institute for Biological Science, Chinese Academy of
Sciences, Shanghai, Peoples Republic of China

2
Laboratory of Medicinal Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences,
Dalian, Peoples epublic of China
Rhizoma Anemarrhenae (Zhimu in Chinese), the dried rhizome of Anemarrhena asphodeloides Bge. (Fam. Liliaceae), is a well-known traditional Chinese medicinal herb and has been used clinically in China for centuries to cure various diseases. However, like other traditional Chinese medicines, the effective constituents of this medicine, especially the assimilation and metabolites in vivo, which are very important to show their effects, have not been systematically studied. In this paper, solid-phase extraction and liquid chromatography-atmospheric pressure chemical ionization mass spectrometry technologies were used to study the constituents absorbed into rat urine and their metabolites after oral administration of Rhizoma Anemarrhenae decoction. A total of 11 compounds, including two xanthones, three of their metabolites and six steroidal saponins, were identified in rat urine sample. They were neomangiferin (1), glucuronide and monomethyl conjugate of mangiferin (2), mangiferin (3), monomethyl conjugate of mangiferin (4), dimethyl conjugate of mangiferin (5), timosaponin N or timosaponin E1 (6), timosaponin BII (7), timosaponin BIII (8), anemarrhenasaponin I or anemarrhenasaponin II (9), timosaponin AII (10) and timosaponin AIII (11). The results would efcaciously narrow the potentially active compounds range in Rhizoma Anemarrhenae decoction, and pave a helpful way for follow-up mechanism of action research.
213
Study On Mango Leaf and Mangiferin Purification and analysis by high performance liquid chromatography of Mangiferin
Xiaoyi Yang, Yulin Tu, Xuemei Zhu, Huifang Xu Hengyang Medical College, Hengyang, P.R. China
In this paper, ethanol re-crystallization technology was applied to the purification of crude mangiferin. High performance liquid chromatography (HPLC) method was established for the analysis of mangiferin. The results showed that the purified mangiferin was HPLC grade. HPLC showed a single mangiferin peak and the same retention time by 3.42min compared with standard sample. Ultraviolet characteristic absorption peak was 220nm. Moreover,It was found that the loss rate was 28.5% after filtration by G6 filtrator while mangiferin being solubilized with DMSO. The actual suppressive concentration that suppressed the duplication of herpes simplex virus type in Hela cells was obtained by correcting the concentration in the cell experiment.
Pulse radiolysis studies of mangiferin: A C-glycosyl xanthone isolated from Mangifera indica
B.Mishra1 ,K.IndiraPriyadarsini1* ,M.Sudheerkumar2 ,M.K.Unnikrishhnan2,H.Mohan1
1
Radiation Chemistry & Chemical Dynamics Division, Bhabha Atomic Research Centre, Trombay,
Mumbai, India
2
College of Pharmaceutical Sciences, Kasturba Medical College, Manipal, India
Pulse radiolysis technique has been employed to study the reaction of different oxidizing and reducing radicals with mangiferin. The reaction of OH radical showed the formation of transient specie sabsorbing in 380-390 and 470-480 nm region. The reaction with specicone-electron oxidants (N3, CCl3O2) also showed the formation of similar transient absorption bands and is assigned tophenoxyl radicals. The pKa values of the transient species have been determined to be 6.3 and 11.9. One-electron oxidation potential of mangiferin at pH9 has been found to be 0.62V vs. NHE. The reaction of e
aq
showed the formation of
transient species with max at 340 nm, which is assigned to the ketyl anion radical formed on addition of e
aq
at carbonyl site. Reactions of one-electron oxidised mangiferin radicals with ascorbic acid have also been
studied.
214
Technology and Application

Summarize of mangiferin products
Zhiping Wang, Jianqin Liang, Jiagang Deng Guangxi Traditional Chinese Medical University, Nanning, P. R. China
Abstract: The mangiferin bulk drug, high purity mangiferin (>98%), was obtained from mango leaf. Even the solubility of mangiferin was very low, it was still increased by 300 folds by inclusion technic. The bulk drug was prepared to 4 kinds of products, including mangiferin tablet, mangiferin injection powder, mangiferin infusion and mangiferin cream. Keywords: Mangiferin; Tablet; Injection Powder; Infusion; Cream; Preparation
Mangiferin is a C-glucosylxanthone from Mangiera indica L. stem bark, leaves, heartwood, roots. The reported pharmacological activities of mangiferin include antioxidant[1], radioprotective[2], antitumor[3], anti-allergic[4], immunomodulatory[5], anti-inflammatory[6], antidiabetic[7], etc.The solubility of mangiferin is low, which limited its pharmaceutical application. In the passed few years, Dengs research team, founded in Guangxi TCM University, have devoted to the investigation of mangiferin, and obtained great achievements. Mangiferin tablet, mangiferin injection powder, mangiferin pills and mangiferin cream are prepared. Some preparations are developed in other countries, such as Vimang[8] extracted from Mangifera indica L. stem bark widely used as a nutritional supplement in Cuba, mangoherpin[9] capsule/cream/gel from Vietnam used as a medicine for herpes virus infections. Mangiferin tablet Mangiferin tablet is used as medicines to treat kings of conditions, such as cough, slight fever, phlegmatic, etc. The research works mainly include several optimizations by experiment design, including formula, procedure parameters. Bulk drug and all the adjuvants were mixed up, then granulated, dried, and punched to tablet. The uncoated mangiferin tablet was coated with flim-coated material to keep all the tablets the same appearance and more stable. After quality standard reseach, pharmacodynamics, toxicology, preclinical study of mangiferin tablet had finished, clinical research I of mangiferin tablet is being conducted now[10].
215

Mangiferin injection powder This injection powder has the same efficacy as mangiferin tablet. Mangiferin has low solubility, which limits its pharmaceutical applications. Inclusion technic is a useful method to improve the water solubility and bioavailability of mangiferin. Inclusion compounds are made for infusion and powder injection use. Especially, the mangiferin inclusion for powder injection use is prepared by frozen drying procedure [11]. Mangiferin pill Mangiferin has wide bioactivities, such as hepatic protection. Drop-pill is a new dosage with high efficiency and bioavailability. Mangiferin pill is useful for treatment of hepatic diseases. But with the pills drawbacks of lower loading drug ratio, it is important to do more pharmaceutical research works. There is one article was issued[12]. Mangiferin cream (5%) Each tube of 10 g mangiferin cream contains Mangiferin 0.5 g. Mangiferin has antihepertic activity with inhibitory effect on the early stages of reproduction of Herpes virus. Mangiferin has an immunostimulating effect on both cellular and humoral immunity link. So mangiferin cream is applied for the treatment of acute and recurrent forms of herpes virus infections of extragenital and genital localizations. Now, mass research works of mangiferin cream are guided by Drug Registration Regulation SFDA Order 28 . References 1.S. Muruganadan, S. Gupta, M. Kataria, et al. Mangiferin protects the streptozotocin-induced oxidative damage to cardiac and renal tissues in rats. Toxicology, 176 (2002) 165. 2.G. C. Jagetia, V. A. Venkatesha. Mangiferin protects human peripheral blood lymphocytes against -radiationinduced DNA strand breaks: a fluorescence analysis of DNA unwinding assay. Mutrition Research, 26 (2006) 303. 3.Y. Maoki, M. Kengo, M. Katayama, et al. The inhibitory effects of mangiferin, a naturally occurring glucosylxanthone, in bowel carcinogenesis of male F344 rats. Cancer Letters, 163 (2001) 163. 4.D.Garcia, M. Escalante, R. Delgado,et al. Anthelminthic and antiallergic activities of Mangifera indica L. stem bark components Vimang and mangiferin. Phytotherapy research, 17(2003)1203. 5.S. Muruganandan, J. Lal, P.K. Gupta. Immunotherapeutic effects of mangiferin mediated by the inhibition of oxidative stress to activated lymphocytes, neutrophils and macrophages. Toxicology ,215 (2005) 57 6.H. S. Bhatia, E. C. Jalil, A. C. P. de Oliveira, et al. Mangiferin inhibits cyclooxygenase-2 expression and prostaglandin E2 production in activated rat microglial cells. Archives of Biochemistry and Biophysics, 477 (2008) 253. 7.S. Muruganadan, K. Srinivasan, S. Gupta, et al. Effect of mangiferin on hyperglycemia and
216

atherogenicity in streptozotocin diabetic rats. Journal of Ethnopharmacology, 97 (2005) 497. 8.G.L. Pardo-Andreu, D.J. Dorta, R. Delgado, et al. Vimang (Mangifera indica L. extract) induces permeability transition in isolated mitochondria, closely reproducing the effect of mangiferin, Vimangs main component. Chemico-Biological Interactions 159 (2006) 141. 9.http://www.bvpharma.com.vn 10.J.G. Deng, Z.P. Wang, W.H.,Shen et al. Study on thin-film coating process for mangiferin tablet. China Pharmacist, 11 (2008) 1140. 11.Zhiping Wang, Jiagang Deng, Qin Wang, et al. Solubility enhancement of mangiferin by HP--CD inclusion technic. Chinese Journal of Hospital Pharmacy. 30(2008)1123. 12.Jiagang Deng, Zhiping Wang, Xuejian Li, et al. Study on processing technology of mangiferin pill. Chinese Traditional Patent Medicine. 30(2008)1070.
217
Study On Mango Leaf and Mangiferin 30 Cases of acute upper Respiratory infection treated with mangiferin tablets
Lilan Qin1, Aiwu Liang2 , Jiagang Deng1
1 2
Guangxi Traditional Chinese Medical University, Nanning, P.R. China Affiliated Ruikang Hospital of Guangxi Traditional Chinese Medical University, Nanning, P.R. China
To assess the effects of mangiferin tablets on acute upper Respiratory infection as well as the safety of the medicine against the disease, the clinical observation selected sixty patients with acute upper Respiratory infection who were then randomly divided into two groups, the treatment group and the control group. Patients in the treatment group, which consisted of 16 men and 14 women, took orally mangiferin tablets three times a day, 3 tablets each time, and this treatment lasted for five days. On the other hand, patients of the control group (15 men and 15 women) took orally Yinqiao Antitoxic Pills (mainly made from honeysuckle flower and forsythia) twice per day, 1 pill each time, and the treatment also lasted for five days. The observation shows that total effective rates were respectively 96.67% and 76.67% for the treatment group and the control group, and significant differences existed between these two rates (P < 0.05). So, mangiferin tablets may be an effective and safe medicine for treating acute upper Respiratory infection.
Optimization of mangiferin extraction process by orthogonal design from Zhimu

Yucun Niu, Changhao Sun, Ying Li, Fuchuan Guo Harbin Medical University School of Public Health, Harbin, PR China
In this paper, the mangiferin extraction process from Zhimu was studied. The orthogonal experimental design was used for the process optimization. The variables factors were ethanol concentration, ethanol volume, bath time, refluxing time. The optimum process conditions was conducted by 60% ethanol, 8 folds ethanol, water bath for 24h, refluxing extract for 2h. The yield was 91.3%. This extraction process of mangiferin from Zhimu was feasible.
218
Study On Mango Leaf and Mangiferin Preparation of mangiferin monosodium salt

Yefei Yuan1 1, 2, Jiagang Deng 1
1 2
Guangxi Traditional Chinese Medical University, Nanning, P.R. China Luzhou Medical College, Luzhou, P.R. China
In this paper, Study of the preparation mangiferin monosodium salt was carried out. Mangiferin was dissolved in water and acetone, then dripped with sodium bicarbonate solution. After filtration, the filtrate was added with acetone. Its structure was characterized by ESI-MS spectra and elemental analysis. Mangiferin monosodium salt was first prepared, and it was crystallized out with 92.8% yield. The synthetic route is practical with high yield.
Solubility enhancement of mangiferin by HP--CD inclusion technic

Zhiping Wang, Jiagang Deng, Qin Wang, Xuejian Li, Huixian Wei School of Pharmacy, Guangxi College of Traditional Chinese Medicine, Nanning, PR China
In the paper, the solubility of mangiferin was increased the by hydroxyp ropy1--cyclodextrin inclusion technic. After optimized the inclusion process and parameters of frozen drying by comparing method, mangiferin inclusion was obtained. The characteristics of mangiferin inclusion were tested by infrared spectrophotometer, UV spectrophotometer, differential scanning calorimetry analysis, phase solubility diagram and solubility determination. The stirred process and frozen drying preparation of HP--CD inclusion compound by the ratio of host/guest molecules 1
1. The solubility of
were fit for the
mangiferin inclusion (tested by 5.99 mg/ml) was 300 times as large as mangiferin bulk drug (tested by 0.111 mg/ml). Hydroxyp ropy1--cyclodextrin inclused mangiferin compound could be prepared by stirred process and frozen drying. Its solubility of the compound was increased significantly.
219
Study On Mango Leaf and Mangiferin Studies on the effects of mango leaf electuary upon influenza
53603 military hospital.
To research into the effects of mango leaf electurary on influenza, the studies selected fifty-four patients infected with influenza who was treated by taking orally mango leaf electuaries with water three times a day, 2 bags each time; the treatment lasted for two days and a follow-up study was done every day. By contrast, 22 patients suffered from the same disease was treated by taking amantadine pills three times per day, 0.2 grams each time, for two consecutive days. The observation shows that Mango leaf electuaries got a total effective rate of 92.6% against influenza, they could eradicated most of the main symptoms (except cough) within 24-48 hours. They reduced fever fast and steadily, without apparent side effects upon the liver function. Besides, they prevented influenza better than amantadine pills (P < 0.01). However, mango leaf electuaries had a weaker effectiveness against influenza than amantadine pills. Anyway, Mango leaf electuaries may be an effective and safe medicine for treating influenza.
Study on mangiferin extraction by air-blasting method

Haibin Huang, Xuejian Li Guangxi Traditional Chinese Medical University, Nanning, P.R. China
The air-blasting method can break plant cell wall, tear plant tissue, make the herbs structure loose. These are beneficial to solvent penetrate and transport inside the herbs, make a substantial increase in surface area. The Mangiferin percolation experiment results are obvious with the yield of 90% and 100% and with the time reduced by half. At the initial 3 h, the yield was 41.8% of air-blasting method compared with yield 27.6% of non-blasting method.
220
Study On Mango Leaf and Mangiferin Study of extraction of Mango Leaf total glucosides tablets
Hua Lu1, Jianmo Huang2
1 2
Guangxi Zhuang Autonomous Region People's Hospital, Nanning, P.R. China Tiantian Le Pharmaceutical Co. Ltd. Guangxi, Liuzhou, P.R.China
In order to improve the quality of Mangguo Zhike tablets, orthogonal design experiment was used for optimization of the extraction of mango leaf. The extraction ratio of mangiferin in mango leaves was the indicator of the experiment. The best conditions of this process are refluxing extraction 3 times by 95% ethanol, extraction for 90min each time. The results show that this process of mangiferin extraction is fit for extraction of total glucosides from mango leaf.
Study on processing technology of mangiferin pills

Jiagang Deng, Zhiping Wang, Xuejian Li, Qin Wang Guangxi Traditional Chinese Medical University, Nanning, P.R. China
In this paper, the formula of mangiferin pills and optimal prescription technology were studied by using single factor method and orthogonal experimental method. Evaluation criteria were pills dropping situation, shape, hardness, roundness, disintegration and mangiferin content. Results showed that coolant was liquid paraffin with a temperature between 6 8 , the ration of drugs and matrix was 1:4, matrix was a mixture of polyethylene glycol6000 and polyethylene glycol4000 (1:8). liquor temperature was 85
89, burette temperature was 8084, and internal diameter of burette was 3mm, while the
dropping distance was about 15cm. It is an optimum formula and process with a high yield. The quality of mangiferin pills was conformity with the standard stated in the Chinese Pharmacopoeia.
221
Study On Mango Leaf and Mangiferin Study on thin-film coating process for mangiferin tablet
Jiagang Deng, Zhiping Wang, Wenhui Shen, Huixian Wei Guangxi Traditional Chinese Medical University, Nanning, P.R.China
In the paper, The conditions of thin-film coating process for mangiferin tablets were optimized. The evaluation indexes were appearance qualified rate, hardness, weight increment, humidity endurance, and dissolution rate. Thin-film coating process conditions were optimized by compare-method and orthogonal method. The best process parameters were that concentration of coated solution was 13%, engine speed was 12r/min, and bed temperature was 45
. The process was feasible and stable.
Study on ultrasonic extraction technics of mangiferin in Anemarrhena asphodeloides Bge

Xiao Yang 1, Juan Dai2, Guangwei Rui 1, Jie Tang1, Xianggui Chen1
1 2
Bioengineering School, Xihua University, Chengdu, P.R. China Department of Lab Medicine, Chengdu Medical College, Chengdu, P.R.China
In this paper, the studies were carried out to optimize the extraction condition of ultrasonic extraction of mangiferin in Anemarrhena asphodeloides Bge. The influencing rules of the mangiferin in Anemarrhena asphodeloides Bge were studied basing on the yield of extracts. Orthogonal experiment was carried out to optimize four variable factors of alcohol concentration, ultrasonic extracting time, ultrasonic power and ratio of solid to liquid. HPLC was used for the determination. The best extraction condition of mangiferin in Anemarrhena asphodeloides Bge were determined, which showed that alcohol concentration was 60%, ultrasonic extracting time was 30 min, ultrasonic power was 40 W and ratio of solid to liquid was 1
70
(by W/V). In this extraction condition, the yield of extracts of mangiferin was 1.85%. Ultrasonic method can save time, be easy to operate, improve extraction rates and save energy.
222
Index
A
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