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DOI:10.3736/ jcim20110914
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Devaraj VC , Krishna BG , Visw anatha GL.Simultaneous
determination of quercetin , rutin and kaempfero l in the
leaf ex tr ac ts o f Moringa olei f era Lam.and Raphinus
sativus Linn.by liquid chromato gr aphy- tandem mass
spectrometry.J Chin Integr Med.2011;9(9):1022-1030.
Devaraj VC , Krishna BG , Viswanatha GL.液相色谱-串
联质谱法测定辣木及萝卜叶的提取物中所含的槲皮素 、
芦丁及山柰酚.中西医结合学报.2011;9(9):1022-
1030.
Received April 23 , 2011;accepted June 26 , 2011;published
online September 15 , 2011.
Full-tex t LinkOut at PubMed.Journal title in PubMed:
Zhong X i Y i J ie He X ue Bao.
Correspondence:Venkatapura C.Devaraj , PhD;Tel:+
91-99-00774992;E-mail:devco lo gy@yahoo.com
Journal o f Chinese Integrative Medicine (J CIM) or Zhong
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ISSN 1672-1977.Published by JCIM Press , Shanghai , China.
Original Experimental Research 实验论著
Simultaneous determinat ion of quercetin , rut in and
kaempferol in the leaf extracts of Moringa oleifera Lam.
and Raphinus sativus Linn.by liquid chromatography-
tandem mass spectrometry
Venkatapura C.Devaraj1 , Burdipad G.Krishna2 , Gollapalle L .Viswanatha3
1.Depar tment of Pha rmacolo gy , Bioneeds Labo rato ry Animals and Preclinical Services , Bangalo re 562111 ,
India
2.Depar tment o f Pharmacognosy , R.R Co llege o f Pharmacy , Bangalo re 560090 , India
3.Depar tment o f Pharmaco log y , Peoples Education Socie ty College o f Pharmacy , Bangalo re 560050 , I ndia
Objective:To develop a rapid and sensitive liquid chromatography-tandem mass spectrometry(LC-MS/MS)method to analyze quercetin(QU), rutin(RU)and kaempferol(KA)simultaneously
in the leaf extracts of Moringa oleifera Lam.and Raphinus sativus Linn.
Methods:Samples were prepared by extracting the leaves of the M .oleifera and R.sativus
by cold-maceration technique using 90% ethanol.Chromatographic separation was operated
with a mixture of 0.2% formic acid in water and acetonit rile at a flow rate of 0.4 mL/min on a
Phenomenex Gemini C18 column with a total run time of 5.01 min.
Results:The MS/MS ion transitionsmonitoredwere 303.03 to 153.1 for QU , 611.1 to 303.1 for
RU , 287.1 to 153.2 for KA and 180.1 to 110.1 for internal standard.The lower limit of quantitation
achieved for QU , RU and KA was 5 ng/mL and the linearity was observed from5 to 2 000 ng/mL.
The correlation coefficients of linear regression analysis were 0.994 6 , 0.995 1 and 0.996 9 for
QU , RU and KA , respectively.
Conclusion:The results indicate that the LC-MS/MS method is fast and sensit ive and may
provide excellent specificity for simultaneous determination of QU , RU and KA in leaf extracts
of M .oleifera and R.sativus .
Keywords:Moringa oleifera ;Raphinus sativus ;plant extracts;quercet in;rutin;kaempferol;
chromatography , liquid;tandemmass spectrometry
·1022· 中西医结合学报 2011年 9月第 9卷第 9期 Jou rnal of Chines e Integrative M edicine , September 2011 , Vol.9 , No.9
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Flavonoids are widely distributed plant metabolites
with st ructur es based on 2-phenylbenzopyr one.
Quercet in (QU), rutin (RU) and kaempfer ol(KA)str ucturally have phenolic groups which
se rve as a sour ce of readily available hydr ogen
atoms , so the fr ee r adicals produced can be delo-
cal ized over the phenolic structure
[ 1-3] (Figure 1).
Flavonoids di ffer from one another in the degree
of unsaturat ion , the pat ter n of hydroxylat ion ,
and the type of sugar linked , and the most common
are the flavones and the flavonols.Flavonoids
such as QU , RU and KA have been repor ted to
have various pharmacological activities like antidiabetic ,
hepatoprotective , cardioprotective , anti-inflammatory ,
antiulcer , antithrombogenic , antineoplastic , antioxidant ,
ant imicr obial and ant ivir al activit ies
[ 4] .Due to
the pharmacological act ivi ties of flavonoids , their
quali tat ive and quanti tat ive determination has
been extensively studied
[ 5] .In this systemat ic
search for bioactive secondary metabolites fr om
plant origin , Moringa oleifera Lam.and Raphinus
sativus Linn.were sele cted for investigat ion.
Figure 1 Structures of quercetin, rutin and kaempferol
Moringa oleifera Lam.(Moringaceae), commonly
known as drum stick , is a multi-purpose tree cultivated
all over India f or its edible leaves and fruits.The
plant is used in tr aditional medicine systems to
treat various diseases.The leaves of the plant have
been reported to have varied source of biological
activities like anti-inflammatory[ 6] , antimicrobial[ 7] ,
hepatoprotective
[ 8] , antitumor[ 9] , ant idiabetic[ 10] ,
antiulcer
[ 11]
and ant ihypertensive
[ 12]
act ivities.The
plant was reported to contain benzyl gucosinolates , β-
sitosterol , glycosides , sugars , alkoloids , flavonoids ,
pr oteins and saponins
[ 13] . Raphinus sativus Linn.(Cruciferae), commonly
known asraddish, is an aroma tic annual herb ,
which is used in t raditional system of medicine to
tr eat var ious diseases.The roots and leaves of
R .sat ivus have been repor ted to possess various
ph armacological act ivit ies like gut-stimula tory
effect
[ 14] , hepatoprotect ive act ivi ty[ 15] , cardio-
protective effe ct
[ 16] , ant ioxidant act ivity[ 17] and
antiurol ithiatic act ivity
[ 18] .The repor ted main
consti tuents of R .sat ivus are alkaloids , proteins ,
polysaccharides , flavonoids , glycosides , and phe-
nolic compounds
[ 19] . I n recent years , high-pe rformance liquid chro-
matogr aphy(HPLC)has been successf ully applied
to the analysis of flavonoids
[ 20 , 21] .Liquid chro-
matogr aphy(LC)is an important tool tha t can be
used quali tat ively as well as quant ita tively for
checking the purity and ident ity of a cr ude drug
and also for quality control of the finished pr od-
uct
[ 22 , 23] .LC coupled with tandem mass spec-
t romete r (LC-MS/MS)is an important tool which
can be used for both qual itative and quant itative
analysis.LC-MS/MS techniques fr equent ly pro-
vide specific , select ive , and sensi tive quant itative
results , often with reduced sample prepar ation
and analysis t ime compared with other commonly
used techniques
[ 24-26] .Nevertheless , compl icated
oper at ion and expensive instruments l imi t the use
of LC-MS/MS for r outine analysis of plant ex-
t racts for quali ty evaluat ion.Sever al chroma to-
gr aphic methods have been documented for deter-
mination of QU , RU and KA in plants , but none
on simultaneous de terminat ion of these phenolics
in leaves by LC-MS/MS method.So f ar , no
·1023·中西医结合学报 2011年 9月第 9卷第 9期 J ou rnal of C hinese Integrative Medicine , Septemb er 2011 , Vol.9 , No.9
repor t has been published on the determinat ion of
QU , RU and KA in M .olei fera and R .sativus
leaves by the LC-MS/MS method.Thus in the
pr esent study an at tempt was made to develop an
LC-MS/MS method f or the simultaneous de termi-
na tion of QU , RU and KA in M .oleifera and
R .sat ivus leaves.
1 Materials and methods
1.1 Chemicals and standards Acetonit rile ,
methanol and formic acid were of high-performance
liquid chromatography (HPLC)grade (Qualigens ,
Mumbai , India).QU(99%), RU(94%)and KA(97%)were purchased from Sigma Aldrich , India ,
and all of the solvents used f or extr action process
and chemicals used f or phytochemical analysis
wer e of analytical gr ade and procured from local
firms.De ionized water was prepar ed by a Mil li-Q
water purificat ion system (Millipore , Milf ord ,
MA , USA).
1.2 Collection and identification of the plant mate-
rial The leaves of M .olei fera and R .sativus
were collected from the Madiwala area of Bangalore ,
I ndia , in the month of December 2009.Botanical
ident ificat ion was done by Prof.Balakrishna
Gowda , Gandhi Krishi Vignana Kendra , University
of Agriculture , Bangalore , India.Voucher specimens(Her bar ium No:UASK-IN-3720 for M .oleifera
and UASK-IN-3250 f or R .sat ivus) have been
kept in our institute museum for future references.
1.3 Preparation of sample solution The fresh
leaves were cleaned , shade dried and reduced to
coarse powder in a mechanical grinder and passed
thr ough sieve No.40.The powdered material
obtained was then subjected to extr act ion by cold-
macer ation using rect ified ethanol (90%) for a
total of 7 d.The extract was fil tered and concen-
tr ated in rotary evaporator under reduced pressure
to yield a thick green ethanolic ext ract.The crude
extract obtained was part ition-fr actionated with
1∶1(v/v , volume r atio)of pe troleum ether and
ethanol(50%)and the mixture was shaken vigor-
ously and kept for about 30 min to make the two
layers separate.The upper layer consisted of
pe troleum e ther , which was removed and concen-
tr ated in a rotary evapora tor to obtain pet roleum
ether fr act ion.The same procedure was repeated
with the r esidue using an equivalent volume of
chloroform and ethyl acetate to obtain the chlor o-
f orm fract ion and ethyl acetate fract ion respec-
tively.The dried ethyl acetate extract was dissolved
in mobile phase containing internal standard ,
filtered through a 0.45μm membrane filter(Millipore)
and finally injected to LC-MS/MS.
1.4 Preparation of standard solution Working
standards were pr epar ed from the stock solut ion(0.5 mg/mL)by dilution with mobile phase con-
taining inter nal standard.The working standar d
concentration ranged from 5 to 2 000 ng/mL.All
standard solutions were filtered through a 0.45 μm
membrane filter (Millipore)and injected directly.
1.5 LC-MS/MS conditions LC-MS/MS experi-
ments were carried out using a MDS Sciex (Foster ,
CA , USA)API 4000 MS/MS equipped with a
Turboionspray
TM
interface at 450 ℃, coupled with
a Schimadzu validat ion and productivity(VP)LC
system(Shimadzu , Japan). Chroma tographic separat ion was performed on
reversed phased stat ionar y phase (Gemini C18
column 50 mm ×2 mm (inner diameter), 5 μm
Phenomenex , Torr ance , CA , USA) with the
inject ion volume of 5 μL.The mobile phase con-
sisted of mixtur e of ace tonitr ile (A)and 0.2%
formic acid in wa ter (B)using a gr adient elution.
The gradient program was set as f ollows:0.01 to
1.80 min 90% to 10%B;1.80 to 2.90 min 10%
to 90% B;2.90 to 5.01 min 90% B.The flow
r ate was set a t 0.4 mL/min.The column oven
temper atur e was kept at 40 ℃.The inter face
between HPLC and MS was atmospher ic pressure
ionization source with the elect rospray inlet oper-
ated in the posit ive mode.The column e ffluent
was le t into MS/MS. The mass par ameters , namely , curtain gas ,
nebulizer gas , auxil lary gas and collision gas were
set at 68.95 , 241.32 , 310.26 and 41.37 kPa ,
respect ively.The compound parame ters , namely ,
decluster ing potent ial (DP), coll ision energy(CE), coll ision exit cell potent ial (CXP) and
entrance potential (EP)for QU , RU , KA and
internal standard were 151 , 51 , 8 and 10 V , 41 ,
31 , 16 and 12 V , 106 , 47 , 14 and 10 V and 53 ,
30 , 12 and 9 V , respectively.De tect ion of the
ions was per formed in the mul tiple re action moni-
toring mode , monitor ing the t ransit ion of the
mass-to-charge ra tio (m/z)303.03 precursor ion
to the m/z 153.1 product ion for QU , m/z 611.1
precursor ion to the m/z 303.1 product ion for
R U , m/z 287.1 precursor ion to the m/z 153.2
product ion for KA and m/z 181.1 precursor ion
to the m/z 110.1 product ion for internal standard.
Quadr apole Q1 was set on low resolut ion whe reas
Q3 was set on unit resolution.The analyt ical data
were processed by the Analyst softwar e (version
1.4.2).
1.6 Calibration curve , limit of detection and limit
of quantification Seven levels of working standard
solut ions were measured in dupl icate.Each cal i-
brat ion curve was const ructed by linear regression
of the average peak area rat io of standar d to
internal standard versus standard concentra tion.
The l imi t of de tect ion (LOD)was the concentr a-
t ion of signal-to-noise r at io of 3 , and limits of
quant ificat ion (LOQ) was dete rmined as the
concentrat ion of signal-to-noise r atio of 10.
2 Results
2.1 Optimization of sample preparation The different
fr actions (that is , petr oleum e ther , chlor oform
and ethyl acetate)of M .olei fera and R .sat ivus
·1024· 中西医结合学报 2011年 9月第 9卷第 9期 Jou rnal of Chines e Integrative M edicine , September 2011 , Vol.9 , No.9
leaves were subjected to preliminary phytochemical
invest igat ions which revealed the presence of
flavonoids in the ethyl acetate f ract ions.Hence
the ethyl ace tate fr actions were selected for LC-
MS/MS analysis. The dried ethyl aceta te extr acts were dissolved
in the chr omatogr aphic mobile phase con taining
the internal standard.Af ter filtering thr ough a
filter paper and a 0.45 μm membr ane filter , the
samples were injected to LC-MS/MS.The com-
par ison of the retention t imes of mass spect ra
with those obtained f rom the standard provided
unequivocal confirmat ion of the ident ificat ion of
QU , RU and KA(Figures 2 to 4).
2.2 Optimization of chromatographic conditions
The feasibility of various mixtur es of solvents
such as ace tonitr ile and methanol and different
buffers such as ammonium ace tate , ammoniun
formate and formic acid with altered flow rates
and different gradient composi tion was tested for
complete resolut ion of QU , RU , KA and inter nal
standard(data not shown).The resolut ion of the
peaks was achieved with a gradient composition
of 0.2%formic acid:acetoni trile wi th a flow rate
of 0.4 mL/min , on a Phenomenex Gemini C18
column , and was found to be suitable for the
determinat ion of elect rospr ay r esponse for QU ,
RU , KA and inter nal standard.
Figure 2 Typical multiple reaction monitoring chromatograms of quercetin(left panel)and internal standard
(right panel)in a)mobile phase , b)reference standard , c)Raphinus sativus
leaf extract , and d)Moringa olei f era leaf extract
·1025·中西医结合学报 2011年 9月第 9卷第 9期 J ou rnal of C hinese Integrative Medicine , Septemb er 2011 , Vol.9 , No.9
Figure 3 Typical multiple reaction monitoring chromatograms of rutin(left panel)and internal standard
(right panel)in a)mobile phase , b)reference standard , c)Raphinus sativus
leaf extract, and d)Moringa olei f era leaf extract
2.3 Optimization of MS/MS detection conditions To opt imize elect rospray ionisat ion (ESI)con-
dit ions for QU , RU , KA and internal standard ,
quadr upole full scans were carr ied out in posit ive
ion de tection mode.During a direct infusion ex-
per iment , the mass spect ra f or QU , RU , KA and
inter nal standar d revealed peaks at m/z 303.03 ,
611.1 , 287.1 and 181.1 , respectively , as protonated
molecular ions [ M +H ] +.Following detailed
opt imizat ion of MS condi tions the m/z 303.03
pr ecursor ion to the m/z 153.08 product ion was
used for quant ificat ion of QU(Figur e 5), for RU
the m/z 611.10 precursor ion to the m/z 303.1
product ion (Figure 6) and for KA the m/z
287.06 precursor ion to the m/z 153.16 product
ion was used for quant ificat ion purpose (Figure
7), and similarly , f or inter nal standar d the m/z
180.1 precursor ion to the m/z 110.1 was used for
the pur pose of quant ificat ion.
2.4 Calibration curve The cal ibr ation curve was
const ructed using seven levels of wor king standard
solut ions (5 to 2 000 ng/mL).The cal ibr ation
standard cur ve h ad a rel iable reproducibil ity and
the calibra tion curve was prepared by determining
the best f it of peak are a rat io against concentr a-
t ion.
·1026· 中西医结合学报 2011年 9月第 9卷第 9期 Jou rnal of Chines e Integrative M edicine , September 2011 , Vol.9 , No.9
·1027·中西医结合学报 2011年 9月第 9卷第 9期 J ou rnal of C hinese Integrative Medicine , Septemb er 2011 , Vol.9 , No.9
·1028· 中西医结合学报 2011年 9月第 9卷第 9期 Jou rnal of Chines e Integrative M edicine , September 2011 , Vol.9 , No.9
3 Discussion
LC-MS/MS is often employed to ident ify and
analyze chemical compounds due to the specificity
and high sensit ivi ty of mass spect rometer , though
ma trix effe ct or ion suppression may consti tute an
obstacle to perform the study.Most of the analyt ical
methods reported , for de terminat ion of QU , RU
and KA requir e a long run time of around 15 to
30 min and have a h igh quant ificat ion limit(10 μg/mL).The presented method with a total
run time of 5 min provided excellent specificity to
detect flavonoids (QU , RU and KA) in leaf
ext racts of M .olei fera and R .sat ivus and was
found to be fast , simple , pr ecise , sensi tive and
accurate.The method described saves 75%of the
analysis time.
4 Competing interests
The authors declare that they have no competing
interests.
REFERENCES
1 Harbo rne JB.Nature , distribution and function o f plant
flavono ids.P rog Clin Biol Res.1986;213:15-24.
2 Robards K , P renzler PD , Tucker G , Sw atsitang P ,
Glover W.Phenolic compounds and their r ole in oxida-
tive processes in fruits.Food Chem.1999;66(4):401-
436.
3 Niko lic KM.Theore tical study of pheno lic antiox idants
pr opertie s in reaction w ith oxygen-cente red radicals.J
Mol S truct.2006;774(1-3):95-105.
4 Tapa s AR , Sakarkar DM , Kakde RB.F lavonoids a s
nutraceuticals:a review.T rop J Pharm Res.2008;7
(3):1089-1099.
5 Ververidis F , T ranta s E , Doug las C , Vo llmer G ,
Kretzschmar G , Panopoulos N.Biotechnology of flavonoids
and o ther pheny lpr opanoid-derived na tur al products.
Par t Ⅱ:Reconstruction of multienzyme pa thw ays in
plants and microbes.Bio technol J.2007;2(10):1235-
1249.
6 Ezeamuzie IC , Ambakederemo AW , Shode FO , Ekwebelem
SC.Antiinf lammato ry effects of Moringa olei f era ro ot
e xtr act.Pharm Bio l.1996;34(3):207-212.
7 C ceres A , Cabrera O , Morales O , Mollinedo P , Mendia
P.Pharmaco log ical proper ties of Moringa olei f era .1:
Preliminar y screening fo r antimic robial activity. J
Ethnopharmacol.1991;33(3):213-216.
8 Pari L , Kumar NA.Hepatopro tective activity of Moringa
olei f era on antituber cular drug-induced liv er damage in
rats.J Med Food.2002;5(3):171-177.
9 Guevara AP, Vargas C , Sakurai H , Fujiwara Y , Hashimo to
K , Maoka T , Kozuka M , I to Y , Tokuda H , Nishino
H .An antitumo r promo ter f rom Moringa olei f era
Lam.Mutat Re s.1999;440(2):181-188.
10 Kar A , Choudhary BK , Bandyopadhyay NG.Comparative
evalua tion o f hypog ly caemic activ ity of some Indian
medicinal plants in alloxan diabetic rats.J Ethnopharmacol.
2003;84(1):105-108.
11 Deva raj VC , Asad M , P rasad S.Effect of leav es and
fruits of Moringa olei f era on gastric and duodenal
ulcer s.Pharm Bio l.2007;45(4):332-338.
12 Gilani AH , Af tab K , Suria A , Siddiqui S , Sa lem R ,
Siddiqui BS , Faizi S.Pharmaco lo gical studie s on hypo-
tensive and spasmo ly tic activities of pur e compounds
fr om Moringa olei fera.Phy tother Res .1994;8(2):
87-91.
13 Goyal BR, Agraw al BB , Goyal RK , Mehta AA.Phyto-
pharmacology o f Moringa olei f era Lam.— an overview.
Indian J Nat P rod Resour.2007;6(4):347-353.
14 Gilani AH , Ghayur MN.Pharmaco log ical basis for the
gut stimulato ry activity o f Raphanus sativus leav es.J
Ethnopharmaco l.2004;95(2-3):169-172.
15 Rifat-uz-Zaman , Ahmad M.Evaluation of hepatoprotec-
tive effects of Raphanas sativus L.J Bio l Sci.2004;4
(4):463-469.
16 Rifat-uz-Zaman.S tudy of ca rdiopro tec tive activity o f
Raphanus sativus L.in the rabbits.Pak J Biol Sci.
2004;7(5):843-847.
17 Ba rilla ri J , Cervellati R, Costa S , Guerr a MC , Speroni
E , Utan A , Iori R.Antio xidant and cho leretic properties of
Raphanus sativus L.sprout(Kaiware Daikon) ex tract.J
Ag ric Food Chem.2006;54(26):9773-9778.
18 Vargas R , Perez RM , Perez S , Zavala MA , Perez C.
Antiuro lithiatic activity of Raphanus sativus aqueous
ex tract on r ats.J Ethnopharmaco l.1999;68(1-3):335-
338.
19 Guti r rez RM , Perez RL.Raphanus sativus(radish):
their chemistry and bio lo gy.ScientificWo rldJournal.
2004;4:811-837.
20 Lunte SM.Structural classification of flavonoids in beverages
by liquid chromatog raphy with ultr av io let-visible and
elect rochemical de tection.J Chromatog r.1987;384:
371-382.
21 Nagel CW.Applica tion o f high perfo rmance liquid
chroma tog raphy to analy sis o f flavonoids and phenyl
propenoids.Cereal Chem.1985;62(2):144-147.
22 Jain A , Lodhi S , Singhai AK.Simultaneous estimation
of quercetin and rutin in Tephrosia purpurea Per s by
high pe rformance thin-laye r chr omatog r aphy.Asian J
T radit Med.2009;4(3):104-109.
23 Nayak PS , Upadhyay A , Dwivedi SK , Rao S.Quantita-
tive determina tion of phyllanthin in Phy llanthus amarus
by high-performance thin layer chromatography.BLACPMA.
·1029·中西医结合学报 2011年 9月第 9卷第 9期 J ou rnal of C hinese Integrative Medicine , Septemb er 2011 , Vol.9 , No.9
2010;9(5):353-358.
24 Mellon FA , Bennett RN , Holst B , Williamson G.Intact
gluco sino late analy sis in plant ex tr acts by pro g rammed
cone vo ltag e elect rospray LC/MS:perfo rmance and
compa rison with LC/MS/MS methods.Anal Biochem.
2002;306(1):83-91.
25 Song L , Mor rison JJ , Bo tting NP , Tho rnalley PJ.
Analy sis o f g luco sinola te s , iso thio cyanates , and amine
degr adation products in vege table ex tracts and blo od
plasma by LC-MS/MS.Anal Bio chem .2005;347(2):
234-243.
26 T ian Q , Rosselot RA , Schw ar tz SJ.Quantitative deter-
mina tion of intact glucosinolates in broccoli , broccoli sprouts ,
Brussels sprouts , and cauliflow er by high-per formance
liquid chroma tog raphy-electro spray ionization- tandem
mass spectrometry.Anal Biochem.2005;343(1):93-99.
液相色谱-串联质谱法测定辣木及萝卜叶的
提取物中所含的槲皮素 、芦丁及山柰酚
Venkatapura C.Devaraj1 , Burdipad G.Krishna2 , Gollapalle L .Viswanatha3
1.Depar tment o f Pharmaco log y , Bioneeds Labo rato ry Animals and Preclinical Ser vices , Bangalo re 562111 , I ndia
2.Depar tment o f Pharmacognosy , R.R Co llege o f Pharmacy , Bangalo re 560090 , India
3.Depar tment o f Pharmaco log y , Peoples Education Socie ty College o f Pharmacy , Bangalo re 560050 , I ndia
目的:通过一种快速 、敏感的液相色谱-串联质谱法测定辣木(Moringa olei f era Lam .)及萝卜(Raphinus
sat ivus Linn.)叶的提取物中所含的槲皮素 、芦丁及山柰酚 。
方法:使用冷浸法(90%乙醇)对辣木及萝卜叶进行提取。提取物与 0.2%甲酸和氰化甲烷混合后以
0.4 mL/min的速度通过 Phenomenex Gemini C18色谱层析柱 ,通过时间为 5.01 min 。
结果:串联质谱法测定的离子转换分别为槲皮素 303.03 ～ 153.1 ,芦丁 611.1 ～ 303.1 ,山柰酚 287.1 ～
153.2 ,内参 180.1 ～ 110.1 。对槲皮素 、芦丁及山柰酚定量测量的最低浓度为 5 ng/mL ,线性分布为 5 ～
2 000 ng/mL。槲皮素 、芦丁及山柰酚的线性回归系数分别为 0.994 6 、0.995 1及 0.996 9。
结论:本研究的结果证明了液相色谱-串联质谱法具有快速 、灵敏的特点 ,对于同时测定辣木及萝卜叶的提取
物中所含的槲皮素 、芦丁及山柰酚有较好的特异性 。
关键词:辣木;萝卜;植物提取物;槲皮素;芦丁;山柰酚;色谱法 , 液相;串联质谱法
·1030· 中西医结合学报 2011年 9月第 9卷第 9期 Jou rnal of Chines e Integrative M edicine , September 2011 , Vol.9 , No.9