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1、三七不同部位及总皂苷提取物的HPLC指纹图谱研究崔翰明,张秋燕,彭亮,路文龙,王阶*中国中医科学院广安门医院(北京市宣武区北线阁5号,100053)摘要:目的建立三七不同部位及总皂苷提取物的HPLC指纹图谱分析方法。方法三七不同药用部位(主根、筋条、剪口、叶、花)和不同规格(20-100头)及总皂苷提取物采用甲醇提取,RP-HPLC/UV法测定,KromasilC18(250mm×4.6mm,5μm)色谱柱;乙腈和水(v/v)线性梯度洗脱;流速:1mL·min-1;柱温:30℃;检测波长:203n
2、m;进样量:20μL。结果三七在80min的色谱图中共确定17个特征峰,主要峰分离良好,经对照品和HPLC/MS判定峰4为三七皂苷R1、峰5为人参皂苷Rg1和Re、峰6为Rf、峰7为Rb1、峰8为Rc、峰9为Rb2、峰10为Rb3、峰11为Rd。以峰4为参照峰(S),计算17个特征峰的相对保留时间和相对峰面积比,各峰精密度、稳定性和测定重现性RSD均小于6.47%。通过相似度软件评价,不同规格三七根(20-100头)和剪口、筋条的指纹图谱近似,相似度大于0.971,但三七根和根茎与花和叶差异很大,
3、相似度分别为0.596和0.564。三七总皂苷提取物因药用部位及生产工艺不同,其主要特征峰的峰面积及比例存在较大差异。结论本方法简便、准确,重现性良好,可用于准确分辨三七不同药用部位及其总皂苷提取物的质量。关键词:三七;高效液相色谱;指纹图谱;三七总皂苷StudyonthefingerprintofdifferentpartofPanaxNotoginsengandtotalsaponinsbyHPLCCUIHan-ming,ZHANGQiu-yan,PENGLiang,LUWen-long,Wa
4、ngJie*Guang’anmenHospital,ChinaAcademyofChineseMedicalScience;Beijing100053,ChinaAbstract:Objective:ToestablishananalysismethodofHPLCfingerprintofthedifferentpartofPanaxNotoginsenganditstotalsaponins.Method:Thedifferentpart(includingroot,rhizome,branc
5、hroot,leaf,flower)anddifferentspecificationsof(20-100piecesper500g)PanaxNotoginsenganditstotalsaponinswereextractedwithmethanol.TheHPLCconditionswereasfollowing:KromasilC18column(250×4.6mm,5μm),acetonitrileandwaterlinearitygradientelution,flowrateat1.
6、0mL·min-1,columntemperatureat30℃,wavelength203nm,injection20μL.Result:Theprimarypeaksin80minHPLCchromatogramofthedifferentpartofPanaxNotoginsengwerewellseparated.The4th,5th,6th,7th,8th,9th,10th,11thpeakwereidentifiedasNotoginsenosideR1,ginsenosideRg1a
7、ndRe,Rf,Rb1,Rc,Rb2,Rb3,RdbycontrolsampleandHPLC/MStechnique,respectively.17characterpeakswereselectedasthefingerprintpeaksofPanaxNotoginseng.Therelativeretaintimeandrelativeareaof17peakswerecalculatedbytakingNotoginsenosideR1peak(4th)asthereferentialp
8、eak.TheRSDsofpreciseandstabilityandrepeatabilitywerelessthan6.47%.ThesimilaritiesoffingerprintswereevaluatedbysimilarityevaluationsystemforchromatographicfingerprintofTCM.Thesimilaritiesoftheroots(20-100piecesper500g)andrhizomeandbranchrootofP