白扁豆总皂苷的含量测定与成分分析

韩君

摘要：[目的]對白扁豆进行提取分离纯化以及含量测定与成分分析。[方法]用石油醚对粉碎的白扁豆脱脂，80%乙醇进行回流提取，正丁醇萃取，浓缩正丁醇得白扁豆总皂苷。白扁豆总皂苷粗提物通过硅胶柱、中压C18色谱柱进行进一步纯化，用薄层监测收集富集皂苷类成分的洗脱液，得到需要成分。采用紫外可见分光光度计（UV）测定，以0.5%香草酸-冰醋酸溶液和高氯酸为显色剂，测定波长为540nm，并采用四极杆-静电场轨道肼高分辨质谱仪（UHPLC-Q-ExactiveOrbitrapMS）在负离子模式下对总皂苷进行成分分析。[结果]白扁豆总皂苷通过硅胶柱、中压C18色谱柱纯化得到较纯的总皂苷。总皂苷UV测定的线性范围为0.0096～0.0192mg/mL（R2=0.9981）;平均回收率为98.15%。白扁豆总皂苷共鉴定出18个化合物。[结论]采用硅胶柱干法上样及中压C18色谱柱分离纯化，该方法纯化度高。UHPLC-Q-ExactiveOrbitrapMS技术与UV法可用于白扁豆总皂苷的定性与定量分析，为后续白扁豆总皂苷生物活性研究提供数据支撑。

关键词：白扁豆;总皂苷;提取纯化;含量测定;成分分析;紫外可见分光光度计;液相色谱-质谱联用

中图分类号R284.1文献标识码A文章编号0517-6611（2021）08-0195-04

doi：10.3969/j.issn.0517-6611.2021.08.051

开放科学（资源服务）标识码（OSID）：

ContentDeterminationandComponentAnalysisofTotalSaponinsinLablabSemenAlbums

HANJun

（NationalandLocalJointEngineeringResearchCenterforKeyTechnologyofChineseMedicinalCompositionGranules，BeijingTcmagesPharmaceuticalCo.，Ltd.，Beijing101301）

Abstract[Objective]Toextract，separateandpurifythelablabsemenalbums，andconductthecontentdeterminationandcomponentanalysis.[Method]Thepulverizedlablabsemenalbumsweredegreasedusingpetroleumether，followedbyrefluxextractionwith80%ethanolandextractionwithn-butylalcohol，andthenthen-butylalcoholwasconcentratedtoobtaintotalsaponinsinthelablabsemenalbums.Thetotalsaponinswereseparatedandpurifiedviasilicagelcolumnandmedium-pressureC18chromatographiccolumn，theeluentofthin-layerredspotswascollectedthroughthin-layermonitoring，andthecomponentsneededwereobtained.Thecontentdeterminationwasrealizedbythecolorimetricmethod.Tobemorespecific，0.5%vanillicacid-glacialaceticacidsolutionandperchloricacidwereusedasthecolordevelopingagents，thedeterminationwavelengthwas540nm，andthecomponentanalysisoftotalsaponinswasimplementedusingthequadrupolerod-electrostaticfieldorbitraphigh-resolutionmassspectrometer（UHPLC-Q-ExactiveOrbitrapMS）underthenegativeionmode.[Result]Thetotalsaponinswereextractedfromthelablabsemenalbumsandpurifiedthroughthesilicagelcolumnandmedium-pressureC18chromatographiccolumn.Thelinearityrangeinthecolorimetricdeterminationoftotalsaponinswas0.0096-0.0192mg/mL（R2=0.9981）;theaveragerecoveryratewas98.15%.Atotalof18chemicalcompoundswereidentifiedfromthetotalsaponinsinthelablabsemenalbums.[Conclusion]Theseparationandpurificationwerecarriedoutthroughthesilicagelcolumndry-typesampleloadingandmedium-pressureC18chromatographiccolumn，withhighdegreeofpurification.TheUHPLC-Q-ExactiveOrbitrapMStechnologyandcolorimetricmethodareapplicabletothequalitativeandquantitativeanalysisoftotalsaponinsinthelablabsemenalbums，andwillprovideadatasupportforthesubsequentresearchontheirbioactivity.