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标题 白扁豆总皂苷的含量测定与成分分析
范文

    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.

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