Separation and purification of alkaloids from Sophora flavescens Ait.

byfocused microwave-
assisted aqueous two-phase extraction coupledwith reversed micellar extraction

abstract

Based on advantages of microwave-assisted extraction (MAE), aqueous two-phase

extraction (ATPE)and reversed micellar extraction (RME), a novel method of extraction and
purification of alkaloids fromSophora flavescens Ait. was put forward by using focused
microwave-assisted aqueous two-phase extrac-tion (FMAATPE) coupled with the reversed
micellar extraction (RME). The aqueous two-phase system(ATPS) consisted of ethanol and
ammonia sulfate was used as extracting solvent, FMAATPE conditionssuch as the
compositions of ATPS, and the extraction temperature, extraction time and solvent-to-
materials ratio were optimized by orthogonal test through HPLC analysis. Then alkaloids
extracted byFMAATPE were further purified with reverse micelles of sodium dodecyl
benzene sulfonate/isooctane/n-octanol. The relative purity of total alkaloids in the product
was improved greatly by RME from 58.30% to88.75% (w/w), and purification recovery was
higher than that of solvent extraction, ion exchange resinand macroporous adsorption resin.
Moreover, the reverse micelles used could be readily recycled byadding salt and water,
keeping better extraction performance with about 80% (w/w) of recovery. Thus,the proposed
method is easy to adjust, operate and enlarge for better application prospect; the purity
ofalkaloids obtained meets the requirements of the compound drug preparation.

1. Introduction

Sophora flavescens Ait. as a typical traditional Chinese medicinehas been commonly

used for the treatment of viral hepatitis, cancer,viral myocarditis, gastrointestinal hemorrhage,
and skin diseasessuch as eczema, colpitis, and psoriasis (Chinese
PharmacopoeiaCommission, 2010). Alkaloids are reported to be its major activeconstituents
like oxymatrine and matrine owing to exhibiting seda-tive, analgesic and other central
nervous system inhibition effectsas well as antipyretic, anti-tumor anti-myocardial activities
(Chenet al., 2010; Ding et al., 2006; Ho John et al., 2009; Huang et al.,2012; Huang and
Chen, 2013; Liang et al., 2012; Xu et al., 2012;Yang and Shen, 2009; Zhang et al., 2011,
2013, 2009; Zhou et al.,2008). As the most important alkaloids, oxymatrine and matrine are
widely utilized not only to evaluate this herb quality, but also tomanufacture pharmaceuticals
such as suppository, capsule, tabletand cream (Chinese Pharmacopoeia Commission, 2010;
Chen et al.,2004; Liu et al., 2010, 2011; Ma and Cheng, 2004; Ma et al., 2012;Ruan et al.,
2010; Wang et al., 2012; Yang and Li, 2012). Moreover,alkaloids also are noticeably used in
agriculture as green pesticidefor insecticidal effect (Li et al., 2006; Lu et al., 2004). Thus,
enor-mous acreages of this herb are grown in the some districts of Chinafor a large quantity
of alkaloids to meet requirement of furtherresearches and extensive applications.
In the traditional extraction and separation of natural products,the plant sample
usually is extracted by organic solvent. This pro-cess is subject to remarkable shortcomings
including the lengthyprocess, high cost of organic solvents, low recovery, and toxic sol-vent
residuals in the products, etc. Aqueous two-phase extraction(ATPE) and reversed micellar
extraction (RME) are widely usedin the separation of biomolecules such as proteins, enzymes
andantibiotics due to simple operation, mild condition, friendly envi-ronment, ease of
scaling-up and low costs (Albertson, 1986; Dekker
 Fig. 1. The scheme of extraction and purification of alkaloids by FMAATPE-RME.

and Leser, 1994; Ding et al., 2015; Guo et al., 2008; Li and Cao,2014; Rosa et al., 2011;
Ruiz-Ruiz et al., 2012; Song and Tang,2004). Aqueous two-phase system (ATPS) is usually
consisted ofeither two incompatible polymers or a polymer and an inorganicsalt in water
above a certain critical concentration. Since shortchain alcohols/hydrophilic solvents and
salts came in to use, ATPEhas simultaneously achieved high extraction yield and high prod-
uct purity in extraction of small molecules from the plants due todemixing effect, low
viscosity and interfacial tension, biocompati-bility and nontoxic. Reverse micelles (RM) are
thermodynamicallystable nanometre droplets containing a certain amount of water,stabilized
in an apolar environment. The constituents could be sol-ubilised in the inner core of a RM (or
be reextracted into a freshaqueous phase using a backward transfer technique), and
isolatedfrom impurities that would be left in the original solution. In termsof the advantages
mentioned above, RME has now been recog-nized as one of the most promising techniques
for separation andpurification of effective constituents.
Recently, ATPE and RME have been applied to extraction of smallmolecules from
biological samples and natural products, holdingor carrying their biologic activities based on
their selectivity, anddemonstrated specific and excellent performance for extraction
ofalkaloids (Chen et al., 2012; Chuo et al., 2014; Dong et al., 2009; Guoet al., 2013; Li et al.,
2011; Liu et al., 2013; Mohd-Setapar et al., 2012;Persona et al., 2015; Ramezani et al., 2015;
Tan et al., 2013, 2014;Wu et al., 2015). Microwave-assisted extraction (MAE) using as oneof
field intensification techniques has been paid more and moreattention in separation of
effective active constituents from naturalproducts due to improving yield, shortening
extraction time, reduc-ing solvent, etc. (Delazar et al., 2012; Liu et al., 2012a). ATPE
canextract effectively alkaloids from Sophora flavescens Ait., removemost of impurities (Liu
et al., 2012c), and attain higher extractionyield when coupled with MAE (Ma et al., 2013;
Zhang et al., 2012;Zhu et al., 2013). Compared with a simple ATPE and other conven-tional
extraction methods, Microwave-assisted aqueous two-phaseextraction (MAATPE) integrated
extraction and purification to one-step procedure, not only intensifying extraction process but
alsopurifying alkaloids based on effects of demixing and microwaveon ATPS. To meet
requirement in pharmaceutical uses especiallycompound drug preparation, further
purification must be devel-oped even if the relative purity of alkaloids could achieve
over70% (w/w) by MAATPE (Zhang et al., 2015). Usually, solvent extrac-tion, ion-exchange
resin and macroporous resin etc can be used topurification of alkaloids, but the procedures
are complicated, longprocess time and large solvent consumption. In our previous work,RME
was found to selectively extract alkaloids from complicatedmatrix under pH 5.0 using the
reverse micelles system of sodiumdodecyl benzene sulfonate (SDBS)/isooctane/n-octanol
(Liu et al.,2012b). Alkaloids in cationic forms were enwrapped into micel-lar droplets while
most of impurities would be left in the aqueoussolution. In view of characteristics and
mechanisms of MAE, ATPEand RME, we integrated MAE with ATPE for rapid extraction of
alkaloids, and subsequently coupled with RME for further purifi-cation. Considering greater
scale of sample pretreatment, focusedmicrowave-assisted extraction (FMAE) is a more
suitable approachto produce more alkaloids than pressurized microwave-assistedextraction
(PMAE) (Delazar et al., 2012; Kaufmann and Christen,2002; Zhang et al., 2012).
FMAE, unlike PMAE limited to 0.1–1.0 gof sample and 5.0–50 mL of solvent, run at
atmospheric pressure,increased the capacity of sample and solvent more than ten times.The
reason is given for allowing FMAE to use larger container forincrease of sample throughput.
Thus, focused microwave-assistedaqueous two-phase extraction (FMAATPE) combining
ATPE withFMAE prior to RME is a better strategy for separation and purifi-cation of
alkaloids from Sophora flavescens Ait. It is would be anattractive, efficient and green
alternative to obtain a relatively highpurity of alkaloids from the herb materials to meet
acquirement of pharmaceutical manufacturing.
In this study, a novel, rapid and effective method for separa-tion and purification of
alkaloids from Sophora flavescens Ait. wasdeveloped by FMAATPE coupled with RME. On
the basis of theresults of PMAE using ATPS of ethanol/ammonium sulfate as sol-vent,
FMAATPE as alternative was established to extract alkaloidsfrom the herb materials in larger
scale of sample pretreatment.Therefore, the factors including microwave power, solvent-to-
materials ratio and the extraction time etc were investigated tooptimize FMAATPE
conditions for improvement of extraction yield.Subsequently, the extract obtained was further
purified by RMEto remove impurities for production of relatively pure alkaloids.Thus,
integrating three technologies together could exhibit theirrespective advantages in extraction
and purification of alkaloidsas compared with the combination of solvent extraction and ion-
exchange separation etc. The method proposed is potential, simpleand easy to operation and
amplification for large scale production

2. Materials and methods

2.1. Materials and reagents

Crude drug of Sophora flavescens Ait. root was obtained fromShanxi Province in
China. After being dried and ground into a finepowder, the samples were placed in
desiccators at room tempera-ture.

Oxymatrine and matrine (purities were ≥98.0%) were purchasedfrom Xi’an Xuhuang
Bio-Tech Co., Ltd. (China). N-oxysophocarpine,sophocarpine and sophoridine (purities
≥98.0%) were boughtfrom Nanjing Zelang pharmaceutical science and technology Co.,Ltd.
(China). sodium dodecyl benzene sulfonate, isooctane andn-octanol(Tianjin Fuchen chemical
regents factory). Acetonitrileand methanol of HPLC grade were purchased from Merck
Darm-stadt Ltd. (Germany). All other chemicals were analytical grade(Guangzhou Chemical
Reagent Factory, China).

2.2. Instruments and apparatus

All extraction experiments were performed on a microwaveextraction system
(PreeKem Scientific Instruments Co., Ltd., China)equipped with a digital timer, power and
temperature controllerand microwave extraction system (Media Electric
Manufacturing,China). HPLC analysis was carried out using Agilent 1200
Infinitychromatograph (Agilent Technologies Co., Ltd. USA)

2.3. The preparation of aqueous two-phase system (ATPS) andreverse micelles system
(RMS)
The ATPS of ethanol/ammonium sulfate was prepared accord-ing to the phase
diagram plotted by Liu et al. (2012c); Zhang et al.(2015). A certain amount of ammonium
sulfate was dissolved indeionized water, and then added ethanol, mixed them by a
vortexstirrer. The ATPS containing ethanol (28.0%, w/w) and ammoniumsulfate (18.0%
w/w) was obtained when the mixture showed twophase separation.
RMS was prepared according to literature (Liu et al., 2012b).Mixed a certain amount
of SDBS and isooctane-n-octanol (4:1, v/v),and dissolved for 10 min by an ultrasonator.
Added a quantity ofwater to the mixture above, heated in an oscillation bath at 25◦Cand
shook at 180 r/min for 2 h, allow to stand up for a night soas to form a reverse micelles
containing 0.05 mol/L of SDBS andW025 of water. Here W0is water content in reverse
micelles, andcharacterized by the following equation:
W0 = nH2O(mol) / nSDBS(mol) (1)

where nH2Oand nSDBSare the molar mass of water and SDBS, respec-tively.

Fig. 2. The apparatus of FMAATPE for extraction of alkaloids from Sophora flavescensAit.

2.4. Separation strategy

The strategy of FMAATPE coupled with RME for extraction andpurification of
Alkaloids from the herb material was planed accord-ing to the following scheme as shown in
Fig. 1, i.e. Alkaloids wereextracted from Sophora flavescens Ait. by FMAATPE with the
ATPSof ethanol/ammonium sulfate, and then purified by RME usingSDBS/isooctane/n-
octanol as a solvent.
2.5. FMAATPE procedure
10 g of herb material (sieved by 80-mesh) and 400 mL of ATPS ofethanol/ammonia
sulfate added to a flask according to the liquid-material-ratio of 40:1, which was then placed
in a microwaveextraction system and equipped with a cooling tube (as shownin Fig. 2). The
extraction was conducted at 85◦C under 420 W for30 min. After cooling, the extract was
filtered to remove the herbresidue. The top phase was not collected until two phases
wereseparated, and removed the solvent by a controlled evaporation toobtain the residues.
The extraction was assessed by the yield (Y) ofalkaloids, which were calculated as follows:

Yield(mg/g) = me / m (2)
where meis the total amount of alkaloids extracted from the herbmaterial (m)

2.6. REM procedure

The residues evaporated was dissolved in HCl solution (pH = 2).Diluted the solution
with water to keep the concentration oftotal alkaloids between 0.2 mg/mL and 0.3 mg/mL,
and adjustedits acidity to pH = 5 with ammonia. Added the equal volume ofreverse micelles
of SDBS/isooctane/n-octanol to the solution above,and extracted for 5 min by
electromagnetic stirring. Then put thereverse micelle layer into the same volume of 1 mol/L
KCl solu-tion for back extraction. Repeated twice, combined the extractsand concentrated.
Subsequently, desalination and crystallizationwere performed by adding 10 mL of methanol.
After filtering, wash-ing and concentrating, dried alkaloids to obtain crud product.
Thepurification was also evaluated by recovery and relative content ofalkaloids, which were
calculated as follows: Recovery(%) = mRM / mTotol × 100%

Relativecontent(%) = We / W × 100% (4)

where mRMis the amount of alkaloids extracted by REM, and mTotalis the amount of
alkaloids in the residue from FMAATPE; WeandW represent the amount of total alkaloids
and the amount of driedresidue respectively.

2.7. Separation of alkaloids by other methods

2.7.1. Solvent extraction

The residues from FMMATPE or PMAATPE were dissolved inHCl solution (pH =
2.0), and extracted three times with chloroform.The aqueous phase was adjust to pH > 10
followed by three timesof extraction with chloroform, combining chloroform extracts,
thecrude product by evaporated at a vacuum evaporator and dried.

2.7.2. Ion exchange resin

The residues dissolved in HCl solution (pH = 2.0) were trans-ferred to a column
packed with 001 × 732 cation exchange resin,flowing through at 2 mL/min of flow rate. After
washed by water,alkaloids were eluted by 2% (w/v) NaOH solution, and concentratedfor
further determination.
2.7.3. Macroporous adsorption resin
The equal amount of residues was dissolved in water, was adjustto pH > 10 with
ammonia. After cooling, the solution was transfer toa column packed with D101
macroporous adsorption resin, washedthe column with water till impurities were not detected
by thinlayer chromatography (TLC). Then alkaloids were eluted by 30%(v/v) ethanol,
concentrated and followed by HPLC analysis

2.8. HPLC analysis

Contents of alkaloids from FMAATPE or RME were determinedby HPLC with a UV
detector at 220 nm using a Phenomenex GemininC18Column (5 _m, 250 mm × 4.6 mm) as
the stationary phase. Themobile phase was made of methanol (A), acetonitrile (B) and
0.1%(w/v) ammonia solution(pH = 10.3)(C). Alkaloids were eluted at30◦C in the following
gradient mode: 0–6 min: 8.5–9.0% A, 9.5–9.0%B and 82% C; 6–11 min: 9.0–16.0% A, 9.0–
16.0% B and 82.0–68.0%C; 11–45 min: 16.0% A, 16.0% B, 68 .0% C. The injection volume
was20 _L. The flow rate was 1.0 mL/min. The obtained chromatogramwas shown in Fig. 3.

3. Results and discussion

3.1. Optimization of FMAATPE conditions

3.1.1. Optimization of FMAATPE conditions with orthogonal test

Before orthogonal test, the single-factor test was first performedconsidering the
factors of the composition of the ATPS, the sizeof matrix materials, the solvent-to-material
ratio, the microwavepower, and the extraction time. The results could be summarizedas
follows: 28% (w/w) of ethanol, 18% (w/w) of ammonium sulfate,80 mesh of the particle size,
the 40:1 of solvent-to-materials ratio,780 W of the microwave power, and 30 min of the
extraction time.

Fig. 3. HPLC Charomatograms of alkaloids before and after RME extraction. 1-Oxymatrine;
2-N-oxysophocarpine; 3-Sophoridine; 4-Matrine; 5-Sophocarpine.
 a It represents total alkaloids, including oxymatrine, N-oxysophocarphine,sophoridine,
matrine and sophocarphine.

Based on the previous work as to ATPE optimized, the composi-tion of ATPS was within
range from 22 to 26% (w/w) of ethanoland from 18 to 22% (w/w) of ammonium sulfate to
achieve higheryield. However, the enlargement of the scale of sample pretreat-ment must be
considered to impact on the effect of microwavewhen using FMAE. The factors of the
microwave power, solvent-to-materials ratio and the extraction time must be
investigatedbecause they would be outstanding and significant on the yield ofalkaloids. Thus,
orthogonal test was employed to further optimizethe extraction conditions according to a
three-variable, three-leveldesign as shown in Table 1. Each extraction was tested in
triplicate,the experimental results and variance analysis also listed in Table 2and Table 3,
respectively.
According to these analytical results, it can be found thatmicrowave power has more
significant effect on the extractionyield of alkaloids among factors. The factors could be
ranked byimportance for total alkaloids as follows: A (microwave power)
 Fig. 4. The pictures before (a) and after (b) extraction of alkaloids from Sophoraflavescens
Ait. by FMAATPE.
>B (solvent-to-material ratio) >C (extraction time). As seen fromTable 2, the factors that
gave the extraction optimum level wereA2B2C2, in other words, the optimum conditions of
FMATPE weremicrowave power of 420 W, extraction time of 30 min, solvent-to-material
ratio of 40:1.

3.1.2. Validation of optimum conditions

To validate of feasibility of FMAATPE, the experiments were con-ducted under the

optimum conditions, triplicate results followsas 75.40, 76.50, 75.38 mg/g of extraction yield
for total alkaloids,and their mean value and the RSD were also 75.76 mg/g and
2.1%,respectively. The validation results demonstrated that adapting L9(33) orthogonal array
scheme could optimize FAATPE conditionseven obtain higher yield than PMAE, reducing
the time and cost,and improving the performance characteristics.
As shown in Fig. 4, the herb powder sample was located betweenthe salt-rich bottom
phase and the ethanol-rich top phase. Itrepeated the fact which the multiphase process
occurred from thesolid-liquid extraction to the liquid-liquid extraction, corroborat-ing the
mechanism reported by literatures (Fan et al., 2006; Paréet al., 1994; Zhang et al., 2015).
Under the presence of ammoniumsulfate, higher conductivity cause microwave a stronger
effect onthe herb cells and the ATPS, leading cells ruptured and improvingmass transfer of
alkaloids in two extraction processes and drivingphases’ separation (Nagaraj et al., 2003).

3.2. Separation of alkaloids by FMAATPE coupled with RME

For further purification, alkaloids could be selectively extractedusing a RMS of

SDBS/isooctane/n-octanol according to optimumconditions (Liu et al., 2012b). Under pH 5.0,
alkaloids in cationicions were wrapped by reverse micelles, and then transferred to theorganic
phase for removal of impurities. Thus, purification of alka-loids extracted by FMAATPE was
performed by RME, meanwhile thecomparative studies was also carried out with PMAATPE
accordingto extraction conditions of the literature (Zhang et al., 2015).
The results in Table 4 showed that three highlights as fol-lows: the extraction yield of
FMAATPE were obviously higherthan that of others; the purity of total alkaloids was
improvedgreatly improvedgreatly from 58.30 to 88.75% by RME; No matter what
FMAATPEor pressurized microwave-assisted aqueous two-phase extraction(PMAATPE)
could help alkaloids to further be purified, thus RMEalso markedly improved the purity of
alkaloids more than 20%(w/w) by selectively extracting cationic alkaloids using
reversemicelles of SDBS/isooctane/n-octanol. The fact that extraction ofalkaloids from the
herb can be intensified by FMAATPE or PMAATPEusing ethanol/ammonium sulfate system
as solvent is proved bythe experiments above. Relatively, FMAATPE would produce
morealkaloids by FMAE device due to option of flask volume, increas-ing at least twenty-
fold of samples. It is worthwhile to note thatwhy FMAATPE in open mode at atmospheric
pressure possessedhigher than PMAATPE. The possible reason for PMAATPE is due
tohigher pressure and temperature that may lead alkaloids decom-posed (Pan et al., 2008).
Fig. 5 illustrated the RME process for purification of alkaloids.It revealed alkaloids
how to be extracted to reverse micelles. Sinceaccepted protons in acid medium, alkaloids
would be convertedinto cationic forms. Accordingly, they were wrapped by micellardroplets,
entering to the organic phase containing isooctane andn-octanol (Liu et al., 2012b). Because
there are hydrophilic waterpools inside reverse micelles, which were surround by hydropho-
bic solvent. As results, acidifying alkaloids not only increased theirpolarity but also drove
them to be dissolved in water pools. Thus,purification of was achieved by extracting
alkaloids to the organicphase while leaving impurities in the aqueous phase. Alkaloidscould
be released from the organic phase by back-extraction whenadding 1 mol/L KCl solution.
With the salt disorganized reversemicelles, purer alkaloids were recovered and collected in
the aque-ous phase. In addition, reverse micelles would also be reused byrecycling.

3.3. Comprison FMAATPE approach with conventional methods

According to the procedures corresponding to the experimen-tal with identical
extracting agent consisting of ethanol and wateras solvent, the comparative study for
extraction and purification ofalkaloids was accomplished by FMAE coupled with solvent
extrac-tion (SE), ion exchange resin (IER) and macroporous resin (MR),respectively. The
results listed in Table 5 demonstrated that appre-ciable differences in extraction efficiency
were observed whenusing the identical content of 28% (w/w) ethanol. In contrast
toconventional extraction methods (Liu et al., 2012a; Zhang et al.,2012; Chen et al., 2012),
FMAATPE greatly improved the extractionyield of alkaloids under the present of ammonium
sulfate. As addingthe salt in the mixture ethanol-water, an ATPS could be formed,i.e. a rich-
ethanol top phase and a rich-salt bottom phase. Underthe action of microwave field, ATPE
processes would be intensifiedincluding extraction and demixing, allowing alkaloids to the
topphase while leaving impurities in the bottom phase. IRE (Gao, 2011;Yu and Wang, 2006),
MR (Li et al., 2013; Chen et al., 2015) and SE(Tong et al., 2001; Yu and Wang, 2006; Zhang
et al., 2010) are com-petent enough to purify alkaloids from a large amount of impurities,but
need much time and many steps such as loading, washing andeluting etc. Although SE is a
relatively simple, selective extraction of

a Here is the percentage of total alkaloids including oxymatrine, N-oxysophocarpine,

sophoridine, matrine and sophocarpine.

Fig. 5. RME process of alkaloids from Sophora flavescens Ait.

 a Here is the percentage of total alkaloids including oxymatrine, N-oxysophocarpine,
sophoridine, matrine and sophocarpine.

a Here is the percentage of total alkaloids including oxymatrine, N-oxysophocarpine,

sophoridine, matrine and sophocarpine.

alkaloids could be realized by adjusting to the suitable acidity. Fromthe results, RME
successfully extracted alkaloids from FMMATPEresidue using SDBS/isooctane/n-octanol as
reverse micelles, andgreatly increased purity of the product as compared with otherthree
methods. It revealed that RME further could remove resid-uary impurities, achieving more
effective purification due to itshigh selectivity. Thus, it proved that FMAATPE coupled with
RMEis an effective alternative to extraction and purification of alkaloidsfrom Sophora
flavescens Ait.

3.4. Recycling of RMS

RMS after back extraction was recycled as follows: added themoderate amount of
water to the collected according to W025 of water content, placed in shaking bath at 25◦C
and 180 r/min,then transferred to a separating funnel, removed aqueous phase,repeated twice.
The replicate use of RMS recycled was conductedfor extraction of alkaloids in the extract
from Sophora flavescensAit. The experimental results listed in Table 6 showed that RMSof
SDBS/isooctane/n-octanol possessed enough ability for recov-ery of alkaloids with 83.98%
and 71.19% (w/w) respectively aftertwo extractions but third extraction down to 63.10%
(w/w). Conse-quently, RMS recycled was used for purification of alkaloids againat least
twice, and keeping their relative content between 84.40%and 82.94% (w/w).

4. Conclusion

In this study, FMAATPE coupled with RME was developedfor extraction and
purification of alkaloids with an ATPS ofethanol/ammonium and a RMS of
SDBS/isooctane/n-octanol,respectively. As integrated FMAE with ATPE into one-step proce-
dure, FMAATPE exhibited great advantages such as intensificationof extraction,
improvement of demixing effect and removal ofimpurities. Under the action of microwave
field, no matter whatFMAATPE or PFMAATPE was superior to the conventional extrac-tion
methods in extraction yield, extraction time and solventconsumption etc. FMAATPE takes a
great advantage of samplehandling size, which is at least twenty times more than that
ofPMAATPE. Additionally, RME showed higher selectivity to furtherseparate alkaloids for
removal of the residual impurities in theextracted from FMAATPE. With FMAATPE
coupled with RME, thepurity of alkaloids was greatly improved from 58.30% and
88.75%(w/w). Moreover, RME used could be recycled at least twice, main-taining recoveries
relative purities of alkaloids between 83.98% and71.19% (w/w). The method proposed
demonstrated a simple, effi-cient and easy scale-up for the selective extraction and
purificationof alkaloids from Sophora flavescens Ait.