高效液相色谱法测定芹菜中芹菜素含量

:(:60171):(1964-),,E-mai:[e-mail]::2008-12-0325220093Vol.25,No.2Mar.2009Determinationofapigenincontentfromcelerybyhighperformanceliquidchromatography(HPLC)王克勤1,2WANGKe-qin1,2罗军武1LUOJun-wu1陈静萍2CHENJing-ping2严志慧1YANZhi-hui1(1.,410128;2.,410125)(1.KeyLaboratoryofTeaScienceofMinistryofEducation,HunanAgricultureUniversity,Changsha,Hunan410128,China;2.HunanInstituteofAtomicEnergyApplicationinAgriculture,Changsha,Hunan410125,China):建立一种测定芹菜中芹菜素含量的高效液相色谱法,测定湖南不同地区的芹菜中芹菜素含量采用高效液相色谱法,色谱柱为KromasilC18,流动相为乙腈和水(3565),流速1.0mL/min,检测波长270nm,柱温为35,进样量:10L,检出限:18ng/mL在10~160g/mL范围内呈良好的线性关系(r2=0.9974),芹菜素平均回收率102%,(RSD=1.91%)该方法操作简便快速分离度,灵敏度高,适用于芹菜素的测定:高效液相色谱;芹菜;芹菜素Abstract:Itwasaimthatthemethodtomeasureapigenincontentofcelery(ApiumgraveolensL.)wasestablishedbyhighperformanceliquidchro-matography(HPLC).TheapigenincontentsofceleryweredeterminedindifferentareainHunan.Thedeterminateconditionwereasfollows:theseparationperformedonKromasilC18column,mobilephasewasaceton-itrile-water(3565,VV),theflowratewas1.0mL/min,andmeasurewavelengthwas270nm,thecolumntemperatureis35.injectedsam-plevolume10L,limitofdetection:18ng/mL.Thecalibrationcurveofapigeninshowedgoodlinearitywithpeakareaintherangeof10~160g/mL,(r2=0.9974),andtheaveragerecoveryratewas102%(RSD=1.91%).Theadvantagesofthismethodarebriefness,speed-iness,wellseparation,andhighsensitivedegree.Keywords:Highperformanceliquidchromatography(HPLC);Celery;Apigenin(ApiumgraveolensL.),,,,(apigenin),,4,5,7-,4,5,73C2C3,[1,2],[3],[4][5,6],[7],,,1材料与方法1.1:65,70%,110(WV),70,2h,;98%:;95%:;:,;:4504MP,Sartorius;:KQ-3200,;:LC-10ATVP,Shimadzu;:R-200,BCHI;:98-I-C,;:AIPHR1-2LD,Christ;:UV-2550,Shimadzu1.21.2.1::KromasilC18(1504.6mm,5m);:(3565);741.0mL/min;35;UV,:270nm1.2.2(1):50mg100mL,(2):0.5,1,2,4,6,8mL25mL,0.45m10,20,40,80,120,160g/mL(3):15mg25mL,,,0.45m1.2.3,200~700nm1.2.410L,(X),(Y)1.2.5,(SN)31.2.61.2.2(40mg/mL)0.5,3,7hRSD1.2.7(80g/mL),7,,RSD1.2.81.2.2(40mg/mL),7,RSD1.2.9(20g/mL)5,0.5,1,1.5,2,2.5mg25mL,0.45m,10L,1.2.1070%,110(WV),70,2h,15mg,2结果与分析2.11,200~700nm,270nm335nm2,,270nm,,270nm,12.20.5,1,2,4,6,8mL25mL,25mL0.45m,,,:Y=32678x-77328,R2=0.9974,:10~160g/mL,1,23,41Table1Peakareaofthecontentofapigeninstandardsample/(gmL-1)10204080120160/Au2521305129481104397260344539893725048021图1芹菜素标准品的全波长光谱扫描图Figure1Ultra-violetabsorptioncurveofapigeninstandardsample图2芹菜素标准品液相色谱图Figure2HPLCchromatogramofapigeninstandardsample图3芹菜提取物样品溶液液相色谱图Figure3HPLCchromatogramofceleryextractionsolutionsample752009年第2期图4芹菜素标准品的液相标准曲线Figure4HPLCstandardcurveofapigeninstandardsample2.3,,(S/N)3,18ng/mL2.4,0.5,3,7h,2,7h,RSD(n=3)0.93%2Table2Resultofstabilitytest(n=3)/h0.537RSD/%/(gmL-1)0.4820.4870.4910.932.510L7,3HPLCRSD(n=7)0.93%2.6,7,4,RSD%1.08%,2.7,0.5,1,1.5,2,2.5mg,3Table3Resultofprecisiontest(n=7)/(gmL-1)RSD/%179.573279.106379.564478.8750.93579.398678.298777.6374Table4Resultofrepeattest(n=7)/(gmL-1)RSD/%10.49220.48230.49640.4961.0850.48660.48970.4945,,102%,RSD(n=5)1.91%2.8,65,70%,110(WV),70,2h,15mg25mL,,,65Table5Resultofcallbackrate/(mgmL-1)/mg/(mgmL-1)/%/%RSD/%(n=5)10.020.50.51899.620.0211.066104.530.021.51.552102.11021.9140.0222.058101.850.022.52.521100.076252:6Table6Measurementresultsofapigenincontentfromcelerybreed/%0.0410.0100.0130.0590.0130.0520.0260.0100.0720.0150.0220.0280.0140.0160.0380.0050.0230.0030.0020.0200.0330.0040.0880.0350.0370.0123结论,,,,,HPLC,[8],[9][10],,0.011%~0.046%0.09%~0.12%,,KromasilC18,(3565),1.0mL/min,270nm,35,:10L,:18ng/mL10~160g/mL(r2=0.9974),102%,(RSD=1.91%),,,,,,,,,,:,0.003%~0.088%,1RomanovaD,VachalkovaA,CipakL,eta.lStudyofantioxidanteffectofapigen,iliteolinandquercetinbyDNAprotectivemethod[J].Neo-plasma,2001,48(2):104.2NarumiS,TakayukiA,MikaeO,eta.lAnt-iandpro-oxidativeeffectsofflavinoidsonmeta-linducedlipidhydroperoxide-dependentlipidperoxidationinculturedhepatocytesloadedwitha-linolenicacid[J].FreeRadicalBiology&Medicine,1999(27):1313.3,,.[J].,2000,20(1):51.HuangZhengming,YangXinbo,CaoWenbin,eta.lExperimentalstudyofQinlingChongiionliverprotection[J].ChineseJournalofHospitalPharmacy,2000,20(1):51.4TsiD,TanBK.Efectsofceleryextractand3-N-butylphthalideonlip-idlevelingeneticallyhypercholester-olaemic(RICO)rats[J].Clin.Exp.PharmacolPhysio,l1996,23(3):2l4~217.5,,,.[J].,2006,28(1):6~9.ZhouHu,iLuXiangyang,TianYun,eta.lAdvancesinstudiesonchemicalconstituentsandpharmacologicalactivitiesofapiumL[J].AminoAcids&BioticResource,2006,28(1):6~9.6HessenauerA,MontenarhM,GotzC.InhibitionofCK2activitypro-vokesdifferentresponsesinhormonesensitiveandhormonerefractoryprostatecancercells[J].Int.J.Onco1.,2003,22(6):126,370.7,,,.[J].,2006,22(3):57~61.WangKeqin,Chenliang,LiuZhonghua,eta.lAdvanceoncelerysynthesesprocessinganditsfunctionalcomponent[J].Food&Ma-chinery,2006,22(3):57~61.8,.HPLC[J].,1997(4):33~34.DaiHua,XiaoLiming.DeterminationofresidueoforganochlorinespesticideinorangebyHPLC[J].Food&Machinery,1997(4):33~34.9,,.[J].,2007,16(5):16~17.LuYingnian,WuKefeng,LiangNianc.iQuantitativedeterminationofapigenininpterissemipinnataL.byHPLC[J].ChinaPharmaceut-ica,l2007,16(5):16~17.10,,,.HPLC3[J].,2006,26(10):1404~1407.