REVIEW JURNAL KIMIA BAHAN ALAM

“Evaluation of Anti-Inflammatory, Anti-diabetic activity of Indian

Bauhinia vahlii (stem bark)”

Disusun oleh :
Nama (NIM)

PRODI FARMASI
SEKOLAH TINGGI ILMU KESEHATAN
BAKTI TUNAS HUSADA
TASIKMALAYA

2019
A. LATAR BELAKANG
Sejak dulu tanaman merupakan sumber yang sangat diperlukan untuk pengobatan. Studi produk alami
bertujuan untuk menentukan nilai obat pada tanaman dengan mengeksplorasi dari pengetahuan ilmiah
yang telah ada, penggunaan tradisional dan penemuan agen berpotensi kemoterapi. Peradangan dianggap
sebagai bentuk fisiologis utama dari mekanisme pertahanan yang membantu tubuh untuk melindungi diri
dari infeksi, luka bakar, bahan kimia beracun, alergen atau rangsangan beracun lainnya. Efek samping
dari obat anti-inflamasi yang tersedia saat ini menimbulkan masalah besar selama penggunaan klinis.
Oleh karena itu, diperlukan pengembangan obat anti-inflamasi yang lebih baru dan lebih ampuh dengan
efek samping yang lebih rendah.
Diabetes mellitus adalah penyakit dengan etiologi yang berbeda. Hal ini ditandai dengan terjadinya
penurunan karbohidrat, protein dan metabolisme lemak yang disebabkan oleh ketidakcukupan sekresi
insulin dalam tubuh. Sekitar 140 juta orang di seluruh dunia menderita penyakit diabetes. Penyakit
diabetes menjadi masalah kesehatan serius di masyarakat negara-negara berkembang, dimana
prevalensinya terus meningkat dan pengobatannya relatif mahal atau tidak tersedia. Strategi alternatif
untuk farmakoterapi modern penyakit diabetes mellitus sangat dibutuhkan saat ini, karena
ketidakmampuan dari terapi modern yang ada untuk mengontrol semua aspek gangguan patologis, serta
besarnya biaya dan kemiskinan untuk banyak penduduk pedesaan di negara berkembang. Oleh karena itu,
bahan tanaman dianggap menjadi sumber alternatif untuk mencari tahu agen baru untuk antidiabetes.
Bauhinia vahlii adalah pohon panjat raksasa hijau yang pernah ada dan penjalar terbesar di India dan
dapat tumbuh memanjang hingga 10-30 m. Batang pohon dapat menebal hingga 20 cm. Pohon ini
mempunyai cabang gemuk ditutupi dengan rambut halus kaku. Daunnya mempunyai ukuran sangat
bervariasi, berbentuk hati kedalam, berwarna hijau, berlekuk tumpul, bulat; mempunyai panjang tangkai
daun 3-6 inch, gemuk. Bunga berwarna putih. Buahnya berupa kulit kayu datar dengan rambut halus kaku,
panjangnya 20-30 cm. Tanaman ini telah dilaporkan memiliki agathisf lavone, flavonoid, asam betulinic,
triterpen, campesterol dan steroid di daun, catechin, asam galat metil ester, benzenoid, mopanol dan 4-O-
metil ester. Tanaman ini juga dilaporkan mengandung kaemferol, quercetin, rutin, asam betulinic,
stigmasterol, sitosterol di daun.

B. RUMUSAN MASALAH
1. Apakah ekstrak kulit batang Bauhinia vahlii memiliki efek anti inflamasi dan efek anti diabetes?
2. Apakah ekstrak kulit batang Bauhinia vahlii dapat diamati dengan skrining fitokimia pendahuluan?
3. Apakah ekstrak kulit batang Bauhinia vahlii dapat diamati efek toksisitas akut?

C. TUJUAN PENELITIAN
Tujuan dari penelitian ini adalah untuk mengevaluasi anti inflamasi, aktivitas anti-diabetes dari ekstrak
kulit batang Bauhinia vahlii menggunakan induksi karagenan pada kaki tikus udem dan Streptozotocin
untuk induksi diabetes. Ekstrak ini juga dipelajari untuk efek toksisitas akut dan skrining fitokimia
pendahuluan.
D. METODE PENELITIAN
Alat yang digunakan dalam penelitian ini adalah soxhlet, penggiling mekanik. Sedangkan bahan yang
digunakan adalah Streptozotocin (STZ), karagenan, petroleum eter, benzene, kloroform, etanol, tanaman
Bauhinia vahlii, mencit albino Swiss (20-25g), tikus wistar pria (150-200 g).
3.1. Persiapan Bahan Tanaman
Kulit Batang Bauhinia vahlii

- dikeringkan selama beberapa minggu

- dihancurkan dengan tangan dan dikeringkan lagi
- digiling menjadi bubuk kasar dengan bantuan penggiling mekanik
- digunakan alat soxhlet dengan bahan bubuk (1.400 g) dan diekstraksi berturut-turut dengan
petroleum eter (60-80 ml) untuk menghilangkan lemak zat aktif
- diekstraksi dengan benzen, kloroform dan etanol 95% untuk meningkatkan kepolaran.
- difiltrasi setiap campuran melalui kertas saring whatmann.

Filtrat (Petroleum eter, benzene, kloroform, dan etanol)

- diuapkan dengan rotary evaporator pada 5 sampai 6 rpm dan pada suhu 40°C (perlakuan ini
menghasilkan getah berkonsentrat sebagai hasil ekstrak kasar)
- dibekukan, dikeringkan dan diawetkan pada suhu 4°C.

Berbagai Ekstrak Kulit Batang Bauhinia vahlii

3.2. Skrining fitokimia

Berbagai Ekstrak Kulit Batang Bauhinia vahlii

- Tes fitokimia kualitatif untuk identifikasi alkaloid, flavonoid, steroid, glikosida, saponin, tanin
dan terpenoid dilakukan dengan prosedur standar.

Hasil

3.3. Penelitian Toksisitas akut (LD50)

Mencit Albino Swiss dari 2 jenis kelamin (20-25 gr)

- dilakukan sesuai metode Karber untuk studi toksisitas akut

- dibagi menjadi enam kelompok dengan setiap kelompok terdiri dari enam hewan
- diperlukan selama 90 hari untuk menentukan LD50 berbagai ekstrak Bauhinia vahlii
- digunakan larutan gum akasia (2%) sebagai alat untuk mempersingkat berbagai ekstrak
- diberikan secara oral pada tikus dengan tingkat dosis berbeda tiap kelompok, yang telah berpuasa
semalaman
- dihitung dan ditetapkan LD50 dari berbagai ekstrak
Dosis 353 mg/kg

3.4. Uji Aktivitas Anti-inflamasi

Tikus

- dibagi menjadi 6 kelompok dengan setiap kelompok terdiri dari enam dan dipuasakan selama 12
jam sebelum percobaan
 - diukur volume dari kaki belakang tikus dengan menggunakan plethysmometer dan kemudian 0,1
mL 1% b/v larutan karagenan disuntikkan secara subkutan ke daerah sub plantar tepat berada di
belakang kaki
- diukur pada 1, 2, 3, dan 4 jam setelah injeksi karagenan
- ditentukan volume kaki tikus (Kosolven (2% larutan gum akasia, P.O), berbagai ekstrak dari
Bauhinia vahlii sebagai suspensi dalam 2% larutan gum akasia (P.O) dan ibuprofen (10 mg/kg,
P.O) diberikan 30 menit sebelum injeksi karagenan. Kelompok yang menerima kosolven sebagai
kontrol. Kaki belakang tikus diukur volumenya secara plethysmometrically sebelum dan sesudah
injeksi karagenan, dengan interval selama 4 jam)
- dinyatakan dengan rata-rata ± SEM dan data dianalisis dengan metode ANOVA dilanjutkan
dengan uji-t test (signifikan jika P < 0,05)

Hasil

3.5. Uji Aktivitas Anti Diabetes

Tikus

- dibuat diabetes dengan injeksi intraperitonial dari Streptozotocin (STZ 60 mg/kg berat badan)
yang dilarutkan dalam buffer sitrat (0,1 M, pH 4,5)
- dikonfirmasi dibetes jika adanya STZ dengan mengukur kadar glukosa darah tikus yang
dipuasakan selama 48 jam setelah injeksi STZ
- dinyatakan diabetes jika kadar gula darah di atas 250 mg/dL
- diambil sampel darah dari ujung ekor pada 12, 24, 36 dan 48 jam setelah pemberian STZ
- diukur kadar glukosa darah pada 2, 4, 8, 12 dan 24 jam setelah pemberian oral Glibenklamid dan
berbagai ekstrak dari Bauhinia vahlii
- ditemukan bahwa adanya produksi hiperglikemia yang stabil setelah 48 jam
- dinyatakan dengan rata-rata ± standard error dan data dianalisis dengan metode ANOVA diikuti
oleh Dunnett’s Multiple Comparison Test (signifikan jika P < 0,05).
Hasil

E. HASIL DAN PEMBAHASAN

Hasil tes fitokimia pendahuluan mengungkapkan terdapatnya alkaloid, flavonoid, fitosterol, senyawa
fenolik, dan glikosida. Sementara karbohidrat, protein, asam amino, gum dan musilago dinyatakan tidak ada
(Tabel 1). Studi toksisitas akut berbagai ekstrak Bauhinia vahlii yang telah dilakukan dan LD50 dari semua
ekstrak terletak antara 3.000-4.000 mg/kg berat badan. LD50 yang di dapat, dalam kisaran dosis yang lebih
tinggi sehingga ekstrak aman dan 1/10 dari rata-rata dari semua ekstrak yaitu 353 mg/kg ditetapkan sebagai
dosis untuk prosedur screening farmakologis.
Aktivitas anti-inflamasi ekstrak petroleum eter, kloroform dan etanol Bauhinia vahlii secara signifikan
menurunkan edema kaki tikus yang disebabkan oleh karagenin pada tikus dengan dosis 353 mg/kg
dibandingkan dengan ibuprofen standar (100 mg/kg) ditunjukkan pada Tabel 2. Efek anti-inflamasi yang
signifikan ini mungkin karena penghambatan setiap mediator inflamasi oleh glikosida atau steroid yang ada
dalam ekstrak.

Hasil efek antidiabetes digambarkan dalam Tabel 3. Aktivitas antidiabetes pada tikus yang induksi
diabetes menggunakan streptozotocin dari tabel ditunjukkan bahwa ekstrak etanol dan kloroform Bauhinia
vahlii pada tingkat dosis 353 mg/kg, menunjukkan penurunan yang signifikan dalam tingkat gula darah 2
sampai 24 jam secara progresif sebanding dengan glibenklamid standar.

Dari evaluasi disimpulkan bahwa ekstrak petroleum eter, kloroform dan etanol Bauhinia vahlii
menunjukkan penurunan yang signifikan pada peradangan tikus dibandingkan dengan obat ibuprofen standar
(P <0,05) dan ekstrak kloroform dan etanol menyebabkan penurunan yang signifikan dalam kadar glukosa
darah dibandingkan dengan obat glibenklamid standar (P <0,05). Hasil ini menunjukkan kemanjuran Bauhinia
vahlii sebagai agen terapi yang efektif dalam pengobatan dari radang akut.

F. KESIMPULAN
Ekstrak petroleum eter, kloroform dan etanol kulit batang Bauhinia vahlii mempunyai aktifitas sebagai
anti inflamasi ditunjukkan pada menurunnya peradangan pada tikus dibandingkan dengan obat ibuprofen, dan
sebagai anti diabetes, ekstrak kloroform dan etanol kulit batang Bauhinia vahlii ditunjukkan menurunnya kadar
glukosa darah tikus dibandingkan dengan obat glibenklamid.

G. REKOMENDASI
Penelitian lebih lanjut ekstrak kulit batang Bauhinia vahlii dijadikan dalam sediaan seperti apa yang
cocok untuk pasien, adanya penelitian lebih lanjut tentang ekstrak ini apakah ada interaksi dengan obat lain,
dan publikasi lebih lanjut bahwa ekstrak kulit batang Bauhinia vahlii mempunyai aktifitas sebagai anti
inflamasi dan anti diabetes.
S1382 Asian Pacific Journal of Tropical Biomedicine (2012)S1382-S1387

Contents lists available at ScienceDirect

Asian Pacific Journal of Tropical Biomedicine

journal homepage: www.elsevier.com/locate/apjtb

Document heading doi: 襃 襃 2012 by the Asian Pacific Journal of Tropical Biomedicine. All rights reserved.

Evaluation of Anti-Inflammatory, Anti-diabetic activityof Indian

Bauhinia vahlii (stembark)
Das Surya Narayan1*, Patro Varanashi Jagannath2, Dinda Subas Chandra3
1
Gayatri College of Pharmacy, Jamadarpali, Sambalpur, Odisha, India
2
Roland Institute of Pharmaceutical Sciences, Berhampur, Odisha, India
3
School of Pharmaceutical Education & Research, Berhampur University, Berhampur-760007, Odisha, India

ARTICLE INFO ABSTRACT

Article history: Objective: To evaluate the anti-inflammatory and antidiabetic property of Bauhinia vahlii (stem
Received 10 August 2012 bark) with preliminary phytochemical profile of the extracts. Methods: The dried whole plant
Received in revised from 11 September 2012 material (1 400 g) was packed in soxhlet apparatus and extracted successively with Pet. Ether (PE)
Accepted 8 December 2012
to defat the drug, petroleum ether was removed from the powdered defatted drug which was then
Available online 28 December 2012
extracted with benzene (BE), chloroform(CE) and 95% of Ethanol (EE) as increasing polarity and
all extracts screened for anti-inflammatory and antidiabetic activity using carrageenan induced
Keywords: paw edema and streptozotacin induced diabetic respectively. The toxicity and phytochemical
Phytochemicals screening were done using standard procedure. Result: The preliminary phytochemical tests
Anti-inflammatory revealed the presence of alkaloids, flavonoids, phytosterol, phenolic compounds, and glycoside.
Antidiabetic While carbohydrates, protein, gums and amino acids were absent. The acute toxicity study
Bauhinia vahlii stem bark of various extracts of Bauhinia vahlii was conducted and dose of 353 mg/kg is fixed for anti-
inflammatory and antidiabetic perperty. The pet ether, chloroform and ethanolic extract of
Bauhinia vahlii significantly decreased the paw edema induced by carrageenin in rats at a dose
of 353 mg/kg comparable to standard ibuprofen (100 mg/kg). Similarly in case of antidiabetic
property, the ethanolic and chloroform extract of Bauhinia vahlii at a dose level 353 mg/kg,
showed significant reduction in blood sugar level from 2 to 24 h in progressive manner comparable
to standard glibenclamide (5mg/kg).

1. Introduction available anti-inflammatory drugs pose a major problem

Plants are indispensible sources of medicine since time
immemorial. Studies on natural product are aimed to
determine medicinal values of plants by exploration of
existing scientific knowledge, traditional uses and discovery
of potential chemotherapeutic agents. Phytochemicals are
used as templates for lead optimization programs, which are
intended to make safe and effective drugs [1].
Inflammation is considered as a primary physiologic
defense mechanism that helps body to protect itself against
infection, burn, toxic chemicals, allergens or other noxious
stimuli. An uncontrolled and persistent inflammation may
act as an etiologic factor for many of these chronic illnesses
[2]. Although it is a defense mechanism, the complex events
and mediators involved in the inflammatory reaction can
easily be induced [2]. The side effects of the currently
*Corresponding author: Das Surya Narayan, Assoc. Prof, (Pharm Analysis & Quality
Assurance) Gayatri College of Pharmacy, Jamadarpali, Sambalpur, Odisha.
Tel: -09437158382
E-mail: suryadas2007@rediffmail.com , suryadas2007@gmail.com
during their clinical uses [3]. Therefore, the development of
newer and more potent anti-inflammatory drugs with lesser
side effects is necessary.
Diabetes mellitus is a group of disorders with different
aetiologies. I t is characterized by derangements in
carbohydrate, protein and fat metabolism caused by the
complete or relative insufficiency of insulin secretion and/
or insulin action [4]. Approximately 140 million people
worldwide suffer from diabetes [5].The disease becomes
a real problem of public health in developing countries,
where its prevalence is increasing steadily and adequate
treatment is often expensive or unavailable[6]. Alternative
strategies to the current modern pharmacotherapy of
diabetes mellitus are urgently needed [7], because of the
inability of existing modern therapies to control all the
pathological aspects of the disorder, as well as the enormous
cost and poor availability of the modern therapies for many
rural populations in developing countries.
Diabetes mellitus is ranked seventh among the leading
 Das Surya Narayan et al./Asian Pacific Journal of Tropical Biomedicine (2012)S1382-S1387
causes of death and third when it’s fatal complications are
taken into account [8]. Traditional preparations of plant
sources are widely used almost everywhere in the world to
treat this disease. Therefore, plant materials are considered
to be the alternative sources for finding out new leads for
hypo- /antihyperglycemic agents.
Bauhinia vahlii[9] is a gigantic climbing ever green tree
and largest creeper in India and can grow up to 10-30
m long. The woody stem can get as thick as 20 cm. The
spreading stout branches are covered with rusty fine hair.
Branchlets[10] densely pubescent and terminating in a pair
of revolute tendrils; young branches, tendrils, petioles,
underside of leaves especially along the nerves and
inflorescence clothed with dense ferruginous tomentum.
Leaves [11] very variable in size, often up to 18 in. diam.,
as broad as long or broader, deeply cordate, 11-15 nerved,
cleft through about 1/3 of the length, sub-coriaceous, dark
green and glabrescent above more or less downy beneath;
lobes obtuse, rounded; petiole 3-6 in. long, stout. Flowers[11]
white, on long slender pedicles, in terminal corymbose or
corymbose racemes.
Fruit [12] is a flat woody pod with fine rusty hairs, 20-30
cm long. The plant has been reported to have agathisf
lavone,flavonoid,betulinic acid,triterpene,campesterol
and steroid in leaves [13] , catechin,gallic acid methyl
ester,benzenoid,mopanol and 4-O-methyl ester[13]. The
plant is also reported to contain kaemferol, quercetin, rutin
, betulinic acid[14] and Quercitrin, Stigmasterol, Sitosterol[16]
in leaves.
The purpose of the present study was to evaluate the anti-
inflammatory, anti-diabetic activity of Bauhinia vahlii stem
bark extracts using carrageenan induced rat paw edema
and Streptozotocin induced diabetic. The extracts were
also studied for its acute toxicity effects and preliminary
phytochemical screening.
2. Materials and methods
2.1 Materials
Streptozotocin (STZ) was purchased from Sigma Chemicals
Co. (St. Louis, MO, USA), Carrageenan was procured from
Himedia laboratories, Mumbai, India. All other chemicals
and solvents(Pet.ether,Benzene,Chloroform and Ethanol)
used were of analytical grade and obtained commercially
from Merck- Limited, India, Mumbai.
2.2. Collection and identification of plant material
The Plant Bauhinia vahlii were collected in the month of
October from Padiabahal, Sambalpur, Odisha , India. The
plant material was taxonomically identified by Dr. (Mrs.) Uma
Devi, Head, Department of Botany, Govt. Women’s College,
Sambalpur, Odisha. A voucher specimen (GWC/B-315/09)
S1383
has been deposited in the Herbarium of the Department of
School of Pharmaceutical Education & Research, Berhampur
University, Berhampur-760007, India for future reference.
2.3. Experimental animals
S wiss albino mice ( 20 - 25 g ) and M ale W istar rats
(150-200 g) were purchased from the animal house of Gosh
enterprises, Kolkata and housed in polypropylene cages
at room temperature with proper ventilation. Prior to the
experiments, mice and rats were fed with standard diet for
1 week in order to adapt to laboratory conditions. They were
fasted over night but allowed free access to water before the
experiment. The experimental protocols were approved by
Institutional Animal Ethics Committee (Reg. No.1339/ac/10/
CPCSEA).
2.4. Preparation of plant material
The whole plant was first sun dried for several weeks,
crushed by hands and dried again. Then the crushed parts
of the plant were ground into coarse powder with the help
of a mechanical grinder. By using the concept of the nature
of solubility and distribution of the active ingredients,
powdered material (1 400 g) was packed in soxhlet apparatus
[17] and extracted successively with Pet. Ether (60-80,) to
defat the drug, petroleum ether was removed from the
powdered defatted drug which was then extracted with
benzene, chloroform and 95 % of E thanol as increasing
polarity. The whole each mixture then underwent filtration
through what man filter paper. The filtrates (Pet. Ether,
Benzene, Chloroform and Ethanol filtrate) obtained were
evaporated by rotary evaporator at 5 to 6 rpm and at 40 °C
temperature. It rendered a gummy concentrates. The gummy
concentrate was designated as crude extract of respective
solvent which was then freeze dried and preserved at 4 °C.
2.5. Phytochemical screening
Qualitative phytochemical tests for the identification of
alkaloids, flavonoids, steroids, glycosides, saponins, tannins
and terpenoids were carried out for all the extracts by
standard procedure [18-21] (Table 1).
2.6 Acute toxicity study (LD50)
Acute toxicity study [22-23] was performed according to
Organisation for Economic Co-operation and Development
was performed as per Karber s method. Swiss albino mice
of either sex weighing between 20-25 g were divided into
six groups with six animals each.and of 90 d were used to
determine LD50 of various extracts of Bauhinia vahlii. The
gum acacia solution (2%) was used as a vehicle to suspend
various extracts. The extracts were administered orally at
different dose levels to group of six mice each, which have
S1384 Das Surya Narayan et al./Asian Pacific Journal of Tropical Biomedicine (2012)S1382-S1387
been fasted overnight. The LD50 values of various extracts are
calculate and dose is fixed as 353 mg/kg.
2.7. Experimental design
Thirty-Six experimental animals were randomly selected
and divided into six groups denoted as Group I, Group II,
Group III, Group IV, Group V and Group VI, consisting of 6
Wistar rats in each group. Each group received a particular
treatment i.e. control, standard and the four doses of the
extract. Prior to any treatment, each rat was weighed properly
and the doses of the test samples and control materials were
adjusted accordingly. Group III to Group VI received the
crude extract orally at the doses of 353 mg/kg of body weight.
Group II received intraperitoneal administration of Ibuprofen
at a dose of 100 mg/kg as standard for anti-inflammatory
study, while Group I was kept as control giving 2% gum
acacia solution in normal saline water.
Similarly forty-two animals were selected and divided
into seven groups denoted as Group I, Group II, Group III,
Group IV, Group V, Group VI and Group VII consisting of 6
Wistar rats in each group. Each group received a particular
treatment i.e. normal control, diabetic control, standard and
the four doses of the extract. Prior to any treatment, each
rat was weighed properly and the doses of the test samples
and control materials were adjusted accordingly. Group IV
to Group VII received the crude extract orally at the doses of
353 mg/kg of body weight. Group III received intraperitoneal
administration of Glibenclamide at a dose of 5 mg/kg-body
weight as standard drug for anti-diabetic study, while Group
II was kept as diabetic control giving 2% gum acacia solution
in normal saline water.
2.8. Anti-inflammatory Activity
The anti-inflammatory activity of Bauhinia vahlii was
studied using acute ( carrageenan induced paw edema )
models of inflammation. The experiment protocols were
approved by the Institutional Animal Ethics Committee prior
to the conduct of the animal experiments (Reg. No.1339/
ac/10/CPCSEA).
This model is based on the principle of release of various
inflammatory mediators by carrageenan. Edema formation
due to carrageenan in the rat paw is biphasic event. The
initial phase is attributed to the release of histamine
and serotonin. The second phase of edema is due to the
release of prostaglandins, protease and lysosome[24,25].
Subcutaneous injection of carrageenan into the rat paw
produces inflammation resulting from plasma extravasations,
increased tissue water and plasma protein exudation along
with neutrophil extravasations, all due to the metabolism of
arachidonic acid[26]. The first phase begins immediately after
injection of carrageenan and diminishes in two hours. The
second phase begins at the end of first phase and remains
through third hour up to five hours.
2.9. Method: Carrageenan-Induced Paw Edema in Rats
A nti-inflammatory activity was evaluated using the
Carrageenan induced rat paw oedema according to the
technique of Winter et al [26-31]. The animals were housed
in cages under standard laboratory condition. They had
free access to standard diet and water. The animal were
divided into 6 groups of six animals each and fasted for 12
h before the experiment. The initial right hind paw volume
of the rats were measured using a plethysmometer and then
0.1 mL of 1% w/v carrageenan solution in normal saline
was subcutaneously injected into the sub plantar region
of the right hind paw. The volume of right hind paw was
measured at 1, 2, 3, and 4 h after carrageenan injection, and
the paw volume was determined. The data were expressed
as paw volume (ml), compared with the initial hind paw
volume of each rat. Cosolvent (2% gum acacia solution, p.o),
various extracts of Bauhinia vahlii as suspension in 2% gum
acacia solution (p.o) and ibuprofen (10 mg/kg, p.o) [25] was
administered 30 min before carrageenan injection. The group
received Co solvent was treated as control.The hind paw
volume was measured plethysmometrically before and after
the carrageenan injection, at hourly intervals for 4 h (Table
2).
Vc-Vt
% inhibition of edema= ( ) 伊100
Vc
Where, VT = mean paw volume of test group
VC = mean paw volume of control group
2.10. Anti diabetic Activities
2.11. Induction of diabetes in rats [32-33]
Rats were made diabetic with an intraperitoneal injection
of Streptozotocin (STZ 60 mg/kg body weight) dissolved in
citrate buffer (0.1 M, pH 4.5). Diabetes was confirmed in STZ
rats by measuring the fasting blood glucose level 48 h after
the injection of STZ. Rats with blood glucose level above 250
mg/dL were considered to be diabetic and were used in this
experiment.
2.12. Estimation of blood glucose
Blood samples were collected from the tail tips at 12, 24,
36 and 48 h after streptozotacin administration and found
that stable hyperglycemia produced after 48 h. The blood
sugar level was measured by digital display glucometer (One
touch -Johnson & Johnson Ltd.). Initial blood sample were
taken before the oral administration of the standard drug
(Glibenclamide) and extracts. The blood glucose levels were
measured at 2, 4, 8, 12 and 24 h after oral administration of
Glibenclamide and various extracts of Bauhinia vahlii (Table
3).
2.13. Statistical Analysis
For Anti-inflammatory screening all experimental results
were expressed as mean依SEM and data were assessed by
 Das Surya Narayan et al./Asian Pacific Journal of Tropical Biomedicine (2012)S1382-S1387
S1385

ANOVA Method followed by student’s t-test. P<0.05 was
considered as statistically significant. While for Anti-
diabetic activity, all results are expressed as mean依
standard error. The data was analyzed statistically using
ANOVA followed by Dunnett’s Multiple Comparison Test
using SPSS 10.0 statistical software. The level of significance
was fixed at 5%.
3.3. Anti-inflammatory activity
The pet ether, chloroform and ethanolic extract of Bauhinia
vahlii significantly decreased the paw edema induced by
carrageenin in rats at a dose of 353 mg/kg comparable to
standard ibuprofen (100 mg/kg) shown in Table 2.
3.4. Antidiabetic activity

3. Result The result of antidiabetic effect depicted in Table 3. From

3.1. Phytochemical constituents of the plant
The preliminary phytochemical tests revealed the presence
of alkaloids, flavonoids, phytosterol, phenolic compounds,
and glycoside. While carbohydrates, protein, gums and
amino acids were absent (Table 1).
the table it is revealed that the ethanolic and chloroform
extract of Bauhinia vahlii at a dose level 353 mg/kg, showed
significant reduction in blood sugar level from 2 to 24 h in
progressive manner comparable to standard glibenclamide.
4. Discussion

3.2. Acute toxicity study Inflammation is a common phenomenon and it is a reaction

The acute toxicity study of various extracts of Bauhinia
vahlii was conducted and the LD50 of all extracts found
to lay between 3 000-4 000 mg/kg body weight. The LD50
is in a higher dose range so the extracts are safe and 1/10
average of all extracts i.e. 353 mg/kg was taken as dose for
pharmacological screening procedure.
of living tissues towards injury. Steroidal anti-inflammatory
agents will lyse and possibly induce the redistribution of
lymphocytes, which cause rapid and transient decrease in
peripheral blood lymphocyte counts to affect longer term
response. B. vahlii belongs to family Caesalpiniaceae is a
common plant of Western Odisha. Phytochemical evaluation
of the various extracts of Bauhinia vahlii reveals the
presence of alkaloids, flavonoids, phytosterol, phenolic

Table 1
Preliminary Phytochemical Investigation of various extracts and powder of Bauhinia vahlii.
Inference
Test
PD PEE BE CE EE
Test for Carbohydrates - - - - -
Test for Gums and Mucilages + - - - -
Test for Proteins and Amino Acid + - - - -
Test for Fixed Oils and Fats + + + + +
Test for Phytosterols + + + + +
Test for Glycosides - - - + -
Tests for Saponins + - - - +
Tests for Flavonoids + + _ + +
Tests for Alkaloids + - - - +
Tests for Tannins and Phenolic Compounds - + + - +

Table 2
Anti-inflammatory activity of various extracts of Bauhinia vahlii (Stem bark).
Mean paw volume (mL) 依 SEM
Treatment Dose (mg/kg) Time in minutes
60 120 180 240
Control(2% gum acacia) - 0.48依0.03 0.78依0.09 0.85依0.12 0.89依0.14
Ibuprofen 0.29依0.07 (39.6) 0.28依0.07 (64.1) 0.24依0.06 (71.80) 0.23依0.13 (74.2)
* * * *
100
Pet.Ether. Ext 0.33依0.13(31.25) 0.41依0.09 (47.4) 0.49依0.01 (42.4) 0.42依0.07 (52.8)
* * *
325
Benzene. Ext 325 0.45依0.09(6.3) 0.73依0.12 (6.4) 0.78依0.15 (8.2) 0.78依0.11(12.3)
Chloroform. Ext 0.36依0.07(25) 0.34依0.09 (56.4) 0.35依0.07 (58.8) 0.32依0.11 (64)
* * *
325
Ethanol. Ext 0.36依0.12 (25) 0.42依0.13 (46.2) 0.57依0.17 (33) 0.50依0.09 (43.8)
* * * *
325
Each value is mean依SEM (n=6), Denotes significant difference compared to control value at P<0.05.
*
S1386 Das Surya Narayan et al./Asian Pacific Journal of Tropical Biomedicine (2012)S1382-S1387

Table 3
Antidiabetic activities of various extracts of Bauhinia vahlii stem bark.
Blood Sugar level in mg/dL
Group Time (h)
2h 4h 8h 12 h 24 h
Dose
I Normal control - 88.4依2.07 87.4依1.12 87.1依0.6 87.6依1.7 87.2依1.03
II Diabetic control 2% Gum acacia solution 288.4依8.06 286.7依9.14 287.1依10.4 289.6依7.6 293.2依5.23
III Glibenclamide 5mg / kg /Day 227.0依4.04 171.0依4.57 139.4依2.64 122.2依2.3 114.0依3.7
* * * * *

IV Pet.Ether Ext 320mg/kg 284.3依7.3 281.2依9.4 280.6依9.2 280.3依5.1 288.8依8.1

V Benzene. Ext 320mg/kg 289.4依5.2 284.0依7.5 285.4依9.3 289.2依9.5 292.4依5.3
VI Chloroform. Ext 320mg/kg 260.4依7.5 246.0依3.5 227.3依8.6 212.7依6.3 269.4依8.3
* * * * *

VII Ethanol. Ext 320mg/kg 268.4依5.8 252.2依3.4 233.4依8.1 194.7依2.7 194.7依2.7

* * * * *

All values are expressed as mean依 SEM, n=6, P<0.05 significant compared to control.
*

compounds, and glycoside. Here anti-inflammatory activity

was performed based on the folk lore information by using
the method,
C arrageenan induced inflammation model for the
estimation of anti-inflammatory effect. The development
of oedima in the paw of the rat after the injection of
Carrageenan is due to the release of histamine, serotonin,
and prostaglandin [38-39]. Stem bark extracts of B.vahlii
showed significant anti inflamatory activity. This significant
anti-inflammatory effect may be due to the inhibition of
any inflammatory mediators by the glycosides or steroids [39]
present in the extract.
Similarly the treatment of diabetes with medicines of plant
origin that proved much safer than synthetic drugs is an
integral part of many cultures throughout the world and
gained importance in recent years. India has a rich history
of using various potent herbs and herbal components for
treating various diseases diabetes [34] and several species
of plants have been described as having a hypoglycemic
activity [35-37].
The anti-inflammatory activity in carrageenan-induce
rat paw edema and antidiabetic activity in streptozotocin-
induced diabetic rats of Bauhinia vahlii were monitored at
a dose level 353 mg/kg for a time interval of 1hr to 4 h and
2h to 24h respectively. From evaluation it was concluded
that the Pet.ether, chloroform and Ethanol extract shows
significant reduction of inflammation as compared to
Ibuprofen (P<0.05) and Chloroform and Ethanol extracts
cause a significant decrease in blood glucose level as
compared to standard drug Glibenclamide (P<0.05).
The present result indicates the efficacy of Bauhinia
vahlii as an effective therapeutic agent in the treatment
of acute inflammations and diabetic and also the present
study authentifies the folk lore information on the anti-
inflammatory and antidiabetic property of the plant Bauhinia
vahlii. Further and detailed studies are in process for the
isolation of active constituent responsible for this property
and to idendification of the possible mechanism for the said
properties.
Conflict of interest statement
We declare that we have no conflict of interest.
Acknowledgement
This study was supported by the grants from AICTE (8024/
RIFD/MOD-75/2010-11), New Delhi, India. The Authors are
extremely grateful to Prof. Ashok Kumar Satpathy , Director
and Prof (Dr).S.K.Mohapatra, Principal, Gayatri college of
Pharmacy,Sambalpur, Odisha,India for providing necessary
facilities to carry out this work.
References
[1] Balunas M J, Kinghorn A D. Drug discovery from medicinal plants.
Life Sci 2005; 78: 431-441
[2] Sosa S, Balicet MJ, Arvigo R, Esposito RG, Pizza C, Altinier GA.
S creening of the topical anti-inflammatory activity of some
Central American plants. J Eth.pharmacol 2002; 8: 211-15.
[3] E David E, Elumalai EK, Viviyan Therasa E, Thirumalai T.
Evaluation of anti-inflammatory activity of the leaf extract of
Wrightia tomentosa. Roem & Schult. J Phar Resh 2010; 3:208-209
[4] Ansarullah, Bharucha B, Dwivedi M, Laddha NC, Begum
R, A Hardikar AA,3 and Ramachandran AV. Antioxidant rich
flavonoids from Oreocnide integrifolia enhance glucose uptake
and insulin secretion and protects pancreatic β-cells from
streptozotocin insult. BMC Complement Altern Med. 2011; 11: 126.
[5] S angameswaran, B and J ayakar, B . A nti-diabetic and
spermatogenic activity of Cocculus hirsutus (L) Diels. Afr J Biot
2007; 6(10):1212-1216.
[6] Paul D D, Tédong L, Asongalem EA, Dimo T, Sokeng S D,
Kamtchouing P. Hypoglycaemic and antidiabetic effect of root
extracts of Ceiba pentandra in normal and diabetic rats. Afr. J.
Trad. CAM 2006; 3 (1): 129 -136
 Das Surya Narayan et al./Asian Pacific Journal of Tropical Biomedicine (2012)S1382-S1387
[7] P areek A , S uthar M . A ntidiabetic activity of extract of
Berberis aristata root in streptozotocin induced diabetic rats.
Pharmacologyonline 2010; 2: 179-185.
[8] Trivedi NA, Majumder B, Bhatt JD, Hemavathi KG. Effect of
Shilajit on blood glucose and lipid profile in alloxan-induced
diabetic rats. Indian J Pharmacol. 2004; 36: 373-76.
[9] Baker in Hook.f. Bauhinia vahlii Wight & Arnott Fl.Brit Ind; 1876.
2:279.
[10]Prodr. Bauhinia vahlii Wight & Arnott.Fl.Pen.Ind; 1834. P. 297.
[11]Parker. Bauhinia vahlii Wight & Arnott, Fl.Punj; 1956; 3. P.180.
[12]Ali,P. Bauhinia vahlii Wight & Arnott.Acta Biol; 1967.8.P.241.
[13]D ugasani S , B alijepalli M K , T andra S , P ichika M R .
Antimicrobial activity of Bauhinia tomentosa and Bauhinia vahlii
roots. Pharmacog maga 2010; 6(23):204-207.
[14]D as S N , D ash P , M istry S , P atro V J , C houdhary N S K .
Antimicrobiological activity of Bauhinia vahlii (Stem Bark) against
selected Human Pathogens. IntRJPharmSci.2010; 01(01): 0036.
[15]Sultana, S, Ilyas M, Kamil M, ShaidaWA. Chemical Investigation of
Bauhinia Vahlii. J Indian Chem Soc 1985; 62(4): 337-338.
[16]H arborne J B . P hytochemical M ethods: A G uide to M odern
Techniques of Plant Analysis.New Delhi: Springer, Rajkamal
Electric Press, 3rd Edn.1998: p.1-32, 40-227.
[17]S hah B N , N ayak B S . E xperimental P harmacognosy.
Jalandhar,India: Vikash & Co; 2008, p.190-200.
[18]S hah C S , Q uadry J S . A T ext B ook of P harmacognosy.
Ahmedabad,India: B.S. Shah Prakashan; 2007-08: 13th Edn, p.83-
86,97.
[19]A li M . T ext B ook of P harmacognosy, CBS P ublishers and
Distributers, New Delhi: , 2003, 2nd Edn. p. 64-65,96-97,139-
40,272,283.
[20]TailangM, SharmaA. Phytochemistry:Theory & Practical.New
Delhi,India:Birla Publications; 2008, Ist Edn.p.199.
[21]Ghosh MN. Evaluation of Analgesic activity. In:Fundamentals
of experimental pharmacology, 2 nd ed., S cientific book
Agency,Calcutta; 2005,P.69-71.
[22]Agrahari AK, Khaliquzzama M and Panda SK. Evaluation of
analgesic activity of methanolic extract of Trapa natans. var
Bispinosa roxb Roots. Journal of Current Pharmaceutical Research
2010; 01: 8-11.
[23]Rahman M, Bachar SC and Rahmatullah M. Analgesic and anti-
inflammatory activity of methanolic extract of acalypha indica
linn. Pak. J. Pharm. Sci. 2010; 23(3): 256-258
[24]Chatpaliwar VA , Johrapurkar A A , Wanjari M.M , Chakraborty R
R , Kharkar V T Indian Drugs 2002; 42; 543- 45.
[25]S undhararajan R and R avichandiran V . A nti-inflammatory
activity of Spermacoce ocymoides Burm.F. Journal of Pharmacy
S1387
Research 2012; 5(1):719-721.
[26]Asuzu I U, Sosa S, Loggia D R. The anti-inflammatory activity of
Icacina trichantha tuber. Phytomedicine. 1999; 4(2): 267-272.
[27]Areej M, Taweel AL, Kadriya S, El-Deeb, Maged S, Abdel K, Jaber
S. GC/MS analysis of the fatty acids of three Clematis species
growing in saudiarabia and their Anti-inflammatory activity.
Saudi Pharmaceutical Journal 2007; 15(3-4):224-227.
[28]S ati S C , S ati M D , R aturi R , B adoni P , S ingh H . A nti-
Inflammatory and Antioxidant Activities of Zanthoxylum Armatum
Stem Bark. Global Journal of researches in engineering: J General
Engineering 2011; 5:86.
[29]Das BN, Saha A, Ahmed A. Anti-inflammatory activity of bark of
Xeromphis spinosa. Bangladesh J Pharmacol 2009; 4: 76-78.
[30]P andurangan A , K hosa R L , S iva E . E valuation of A nti-
inflammatory and Analgesic Activity of Root Extract of Solanum
Trilobatum Linn. Iranian Journal of Pharmaceutical Research
2008; 7 (3): 217-221.
[31]V aiyapuri S , D ey A , B alasundaram, B alasubramanian R .
Antihyperglycemic, Antihyperlipidemic and Antioxidant Effect
of Phyllanthus rheedii on Streptozotocin Induced Diabetic Rats.
Iranian Journal of Pharmaceutical Research 2008; 7 (1): 53-59.
[32]M ishra SB , V erma A , M ukerjee A , V ijayakumar M . A nti-
hyperglycemic activity of leaves extract of Hyptis suaveolens L.
Poit in streptozotocin induced diabetic rats. Asian Pacific Journal
of Tropical Medicine. 2011; 689-693.
[33]Yeh GY, Eisenberg DM, Kaptchuk T J, Phillips RS. Systematic
review of herbs and dietary supplements for glycemic control in
diabetes. Diabetes Care 2003; 26: 1277-1294.
[34]Verspohl EJ. Recommended testing in diabetes research. J Planta
Med 2002; 68: 581-590.
[35]De Sousa E, Zanatta L, Seifriz I, Creczynski-Pasa TB, Pizzolatti
M . G , S ypoganicz B , S ilva FRMB . H ypoglycemic effect and
antioxidant potential of kaempferol-3,7-0-(ἀ)-dirhamnoside from
Bauhinia forficate leaves. J Nat Prod 2004; 15(3-4): 829-832.
[36]Colca J R. Insulin sensitizers may prevent metabolic inflammation.
J Biochem Pharmacol 2006; 72: 125-131.
[37]G e o r g e w i l l O A , G e o r g e w i l l U O , N w a n k w o a l a R N P .
Antiinflammatory effects of Morninga oleifera lam extract in rats.
Asian Pac J Trop Med 2010; 3(2): 133-135.
[38]G eorgewill OA , G eorgewill UO . E valuation of the anti-
inflammatory activity of extract of Vernonia Amygdalina. Asian
Pac J Trop Med 2010; 3(2): 150-151.
[39]Mohamed Saleem TK, Azeem A K, Dilip C, Sankar C, Prasanth
NV, Duraisami R. Anti-inflammatory activity of the leaf extacts of
Gendarussa vulgaris Nees.Asian Pac J Trop Med 2011; 147-149.