RP—HPLC测定复方当归妇康胶囊中丹参酮ⅡA的含量（英）

RP—HPLC测定复方当归妇康胶囊中丹参酮ⅡA的含量（英） RP—HPLC测定复方当归妇康胶囊中丹参 酮?A的含量(英) 2005年4月 第27卷第4期 中成药 ChineseTraditionalPatentMedicine April2005 V0J.27No.4 参考文献 [1]宋立人,洪恂,丁绪亮,等主编.现代中药学大辞典(上册) [M].北京:人民卫生出版社.1231. [2] [3] 封锡兰,徐绥绪.苦碟子中新黄酮苷(II)[J].中国药物化学 杂志,2000,IO(2):143. 封锡志,徐绥绪,岳大彪,等.苦碟子中新黄酮苷[J].中国药 物化学杂志,2000,9(3):134—136. RP—HPLC测定复方当归妇康胶囊中丹参酮?A的含量(英) 王新春,于志庚,陈卫军,胡志林 (新疆石河子大学医学院第一附属医院,新疆石河子832008) 关键词:HPLC;当归妇康胶囊;丹参酮?A 摘要:目的:建立HPLC法测定复方当归妇康胶囊中丹参酮IIA含量的方法.方法:采用ODS柱,以甲醇.水(80:10)为 流动相,检测波长为270nln,流速为1.0mL?min,.结果:丹参酮IIA在0.01,0.16Ixg,范围内呈良好线性关系,丹参 酮?A平均回收率99.73%,RSD为2.91%(=5).结论:该方法简便,分离完全,结果可靠,适用于该制剂的质量控制. 中图分类号:R927文献标识码:A文章编号:1001—1528(2005)04-0424-03 DeterminationoftanshinonelIAinCompoundDangguifukangCapsulebyRIP- H]f.LC WANGXin-chun,YUZhi—geng,CHENWei-jun,HUZhi-lin (TheFirstHospitalaffai~atoMedicalCollege,ShiheziUniversity,Shlhezl832008,China) KEYWORDS:tanshinoneIIA;DangguifukangCapsule;HPLC ABSTRACT:A:ToestablishaHPLCmetIlodf0rthedeterminationoftanshinone1IAinCom poundDanguifu— kangCapsule.METHODS:Analyticcolumn:ReliasilC18(5m,250mm× 4.6mln),protectivecolumn:(5 m,12.50mm×4.6mm);mobilephase:methnaol— ultrapurewater(80:20);detectivewavelengthwasat270 nm;temperaturewasat30.C;theflowratewasabout1.0mL?min,.REsULTS:,I'}lestandardcurveshowed agoodlinearitywithintherangeof1×10, -0.16Ixg./-=0.9995.Theaveragerecoverywas99.73%and脚 was2.91%.CoNCLUSIoN:Themethodisfeasible.andsimpletooperateandsuitablefortheq ualitycontrolof CompoundDangguifukangCapsule. CompoundDangguifukangCapsuleispreparedby ourhospital,whichiscomposedofseveraltraditional ChinesemedicinessuchasRadixAngelicasinensis, RadixSalviaeMiltiorrhiaze,andRadixAstragali,and soon.Thispreparationhasbeenusedintheclinicfor thetreatmentandpreventionofmenoxenia,painful menstruation,menoschesisandothergynecologicaldis— eases ( , whichwasprescribedonthebaseofthetheo— ryofTCMandresearchedbyapplyingtheadvanced modelTlpharmaceuticstechnology.Theresultsfrom morethantenyearsclinicalexperimenthavebeensat— istiedwitIlitssmallsideeffectandcurativeeffective— ness.TanshinonelIAisonemaioringredientofRadix SalviaeMihiorrhiaze,Ithasstrongerbiologicalac— tiont【 , so,inthispaperthemethodfordetermination oftanshinonelIAinCompoundDangguifukangCap— sulebyHPLCisreported. 1ApparatusHP1100HPLC,HPRev.A.05.01 I~ttareceived:2004-01-02 Support:TheresearchwassupportedbyShiheziUniversityofNaturalScienceFundCommitt ee,itemnumber:2001-822574 Biography:WANGXin-ehun(1969 一),female,masterdegree,associatedirectorpharmacist.E-mail:wxc69@sohu.corn. 424 2005年4月 第27卷第4期 中成药 ChineseTraditionalPatentMedicine April2005 Vo1.27No.4 [273]chemicalworkstation.1314A—uvvariable wavelengthdetector,electronicscales,inprecisionof oneoftenthousand,modelAE-200(Swiss),ultra— soniccleaningwashmachine,modelKS-5000(Ning— bo,China). ReagentsCompoundDangguifukangCapsule(from TheFirstHospitalAffiliatedtoMedicalCollege,Shihe— ziUniversity),tanshinonelIA(boughtfromNational InstituteforControlofPharmaceuticalandBiological Product,batchnumber0765-200011andforuseofde— termination),otherreagentsandsolventswereA.R. grade. 2Experiment ChromatographicconditionsAnalyticColumn:Re— liasilC18(5Ixm,250mm×4.6Inln),protectivecol— umn:(5Ixm,12.50mm×4.6mm);mobilephase: methanol—water(80:20);detectivewavelengthwasat 270nm;temperaturewasat30.C;theflowratewas 1.0mL?min,:therelativeretentiontimewas17 min;theinjectvolumewas10L. 2.1AssaypreparationGrindthecontentsofnot lessthanCompoundDangguifukangCapsulestoafine powdertopassthrougha40一meshscreen.Transferan accuratelyweighedportionofthepowder,equivalentto about2.0g.toSoxhletextractor,add50mLofe— thertoflaskandextractcontinuouslyfor3hinawater bathmaintainedatthetemperatureof50.C.Evapo— rateethersolutiononawaterbath,controlledatthe temperaturethatwillnotcausetheethersolutiontoboil over,withtheaidofastreamofnitrogen,completethe evaporationremovingthelasttracesofetherwithoutthe applicationofheat,immediatelyswirltodissolvethe residueinsmallportionsmethano1.Transfertoa50 mLvolumetricflask.dilutewithmethanoltovolume, mix.Filterthesolutionthrougha0.45Ixmmicropore filtermembrane,discardingthefirst5mLofthefil— trate.Pipe1mLofthesubsequentfiltratetoacentri— fugetube,centrifugeatabout8000rpmfor4min. IIsetheclearsolutionsoobtaiBedasthetestsolution. 2.2BlanksamplepreparationUsethesamees— tablishedmethodforassaysampletotreatthecapsule withoutRadixSalviaeMihiorrhiage. 2.3Standardcurveoftanshinone1IADissolve anaccuratelyweighedquantityoftanshinonelIA,e— quivalenttoabout12.5mg,ina25mLbrownvolu— metricflask.addmethanolandmixtoobtainastock solutionhavingaknownconcetrationofabout0.50mg oftanshinonelIApermL.Separatelyinjectaliquotof 2O,4O,100,150,200,250,300,320IxL,respec— tively,ofstocksolutionpreparationintoseparate10 mLbrownvolumetricflask,dilutewithmethanolto volume,andmix.Injectequalvolumesoftheprepared solutionintothechromatograph,recordthechromato? grams,andmeasuretheresponsesforthemajorpeaks. Accordingtotherecordedchromatogram.thepeakare— as(Y)withstatisticaltreatmentasafoundationofthe amount(X)gavethegoodlinearityforweighttanshi— none11A.intherangof0.01-0.16Ixg.X=O.078Y 一 0.5793.r=0.9995. 2.4PrecisionInjection,continuouslyoneofthe standardsampleandassaysample10IxL,respective? ly,for5timesandmeasuringthepeakareasofTanshi— none11A.theRSDofstandardsampleandassays枷一 piewere0.84%and2.12%: 2.5Repeatabilityweighup5portionsofsample (batchnumber.-030315),preparingthetestsolution asdirectedundertheprocedure2.2,theinjectvolume ofsamplein10IxLaccordingtothechromatogramre— corded.theRSDwas3.O3%. 2.6SolutionstabilityOneofthestandardsamples thatwasplaced,atroomtemperaturefor1,2,3,4,5 h.respectively,wasinjected.rI1hepeakareaswereba? sicallyunchangeableandRSDwas1.62%.indicating thatthesolutionwasstableatleastfor5h. 2.7RecoveryUntreatedcapsulesampleswere mimedtopassthrough40meshscreenandweighted accuratelyup5portionofsample,everyportionwase— quivalenttoabout1.Og.Separatelyeveryportionof samplehomogenizedwith1.5mLstocksolutionprepa— ration,thenprocessedaccordingtothe2.2procedure. rI1heresultswereobtainedwithaveragerecovery 99.73%.andRSDwas2.91%(n=5). 2.8FeasibilityofmethodInjectionblanksample. oneofthestandardsamplesandassaysample10IxL, resrectively,asshowninFigure1,nointerferences werefound. 425 2005年4月 第27卷第4期 中成药 ChineseI'raditionalPatentMedicine April2005 V01.27No.4 A B Fig.1ItPLCchromatogramsoftanshinonelIA(A)Com- poundI)angeifukangCapsule(B)blanksample(C) 2.9Determinationofsample Weighup3batchesofsamples,preparingthetest solutionasdirectedundertheprocedure2.2,Sepa- ratelyinjectequalvolumesintothechromatograph,re— cordthechromatograms,andmeasuretheresponses 426 forthemajorpeaks.Calculatethequantity,inmg,of tanshinoneIIAineachofthecapsule.Themeasured contentswerelistedinrIle1. Table1Tanshinone?AinCompoundDangulf-nkan~ Capsule(,l=3J 3Discussion 3.1TheselectionofthewavelengthOptimizethe absorbanceofassaypreparationatabout223m,254 nm,270am,determine270nnlasthewavelengthof test,almostwithoutinterferingofothercomponents. 3.2TheelectionofextractingmethodOptimize severaldifferentextractingmethodssuchasultrasonic bath,waterbathataconstanttemperature,and Soxhletextracting,accompanyingwithdifferentextrac— tingsolventsincludingmethanol,ethanol,amixtureof methanolandwater,andether,determinetheextrac— tingmethodaccordingtotherecordedchromatogram, asshowninassaypreparation. 3.3Thereportedmethodissimple,reliabletodeter- minetanshinoneIIAinCompoundDangguifukang CapsuleandsuitableforthequalitycontrolofCorn— poundDanguifukangCapsule. [1]WangJM.PharmacologyofChineseMateriaMedica(中药药理 学)[M].Shanghai:ShanghaiScienceandTechnologyPublishing House.1985:115. [2]ZhengHZ.ApplicationandModemResea~hofTraditionalChi- n.cseMedicine(中药现代研究与应用)[M].2nded.B嘶iIlg: XueyuanPublishingHouse.1997.10:1093. [3]uuY,XiePD,WangBQ.Methodonevaluationofthequality ofMihiorrhizaeanditspreparation[J].Ch/naJChinMaterMed (中国中药杂志),1990,15(3):31.