白扁豆總皂苷的含量測定與成分分析

韓君

摘要：[目的]對白扁豆進(jìn)行提取分離純化以及含量測定與成分分析。[方法]用石油醚對粉碎的白扁豆脫脂，80%乙醇進(jìn)行回流提取，正丁醇萃取，濃縮正丁醇得白扁豆總皂苷。白扁豆總皂苷粗提物通過硅膠柱、中壓C18色譜柱進(jìn)行進(jìn)一步純化，用薄層監(jiān)測收集富集皂苷類成分的洗脫液，得到需要成分。采用紫外可見分光光度計（UV）測定，以0.5%香草酸-冰醋酸溶液和高氯酸為顯色劑，測定波長為540nm，并采用四極桿-靜電場軌道肼高分辨質(zhì)譜儀（UHPLC-Q-ExactiveOrbitrapMS）在負(fù)離子模式下對總皂苷進(jìn)行成分分析。[結(jié)果]白扁豆總皂苷通過硅膠柱、中壓C18色譜柱純化得到較純的總皂苷。總皂苷UV測定的線性范圍為0.0096～0.0192mg/mL（R2=0.9981）;平均回收率為98.15%。白扁豆總皂苷共鑒定出18個化合物。[結(jié)論]采用硅膠柱干法上樣及中壓C18色譜柱分離純化，該方法純化度高。UHPLC-Q-ExactiveOrbitrapMS技術(shù)與UV法可用于白扁豆總皂苷的定性與定量分析，為后續(xù)白扁豆總皂苷生物活性研究提供數(shù)據(jù)支撐。

關(guān)鍵詞：白扁豆;總皂苷;提取純化;含量測定;成分分析;紫外可見分光光度計;液相色譜-質(zhì)譜聯(lián)用

中圖分類號R284.1文獻(xiàn)標(biāo)識碼A文章編號0517-6611（2021）08-0195-04

doi：10.3969/j.issn.0517-6611.2021.08.051

開放科學(xué)（資源服務(wù)）標(biāo)識碼（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.