富硒堇叶碎米荠中硒化合物的形态分析与鉴定  

Detection and Identification of Selenium Compounds in Selenium-enriched Cardamine Violifolia

作　　者：叶梅 虞锐鹏[1] 丛欣 陈尚卫[1] 朱松[1] YE Mei;YU Rui-peng;CONG Xin;CHEN Shang-wei;ZHU Song(State Key Laboratory of Food Science and Technology,Jiangnan University,Wuxi 214122,China;Enshi Se-Run Health Tech Development Co.,Ltd.,Enshi 445000,China)

机构地区：[1]江南大学食品科学与技术国家重点实验室,江苏无锡214122 [2]恩施德源健康科技发展有限公司,湖北恩施445000

出　　处：《分析测试学报》2022年第1期100-107,共8页Journal of Instrumental Analysis

基　　金：国家自然科学基金面上项目(32172197);食品科学与技术国家重点实验室自主科研课题(SKLF-ZZA-202106)。

摘　　要：该文以富硒堇叶碎米荠为研究对象,通过酶解法提取其中的硒化合物,采用高效液相色谱-电感耦合等离子质谱(HPLC-ICP-MS)定量分析已知的硒化合物;对于未知的硒化合物,将酶解液经3 kDa超滤离心管浓缩除杂、冻干后,用初始流动相复溶,选取Kinetex F5超高效液相五氟苯基柱(100 mm×2.1 mm,2.6µm),以0.1%(体积分数)甲酸-水溶液和0.1%(体积分数)甲酸-乙腈溶液为流动相进行梯度洗脱,通过超高效液相色谱-电喷雾电离源高分辨串联质谱(UPLC-ESI-Triple TOF MS),利用硒独特的同位素模式、一级质谱(TOF MS)和二级质谱(TOF MS/MS)进行筛选与鉴定。结果表明:HPLC-ICP-MS定量分析了堇叶碎米荠中的5种硒化合物,分别为硒代胱氨酸(SeCys_(2))、甲基硒代半胱氨酸(MeSeCys)、硒代蛋氨酸(SeMet)、四价硒[Se(Ⅳ)]、六价硒[Se(Ⅵ)];UPLC-ESI-Triple TOF MS鉴定了其他15种硒化合物,包括7种已报道和8种未报道的硒化合物,并提出了C_(4)H_(7)NO_(2)Se、C_(5)H_(8)O_(2)Se和C_(7)H_(9)NO_(2)Se 3种未报道硒化合物的结构。In this article,selenium(Se)compounds were extracted from Se-enriched Cardamine Vio⁃lifolia(C.Violifolia)by enzymolysis method,and the known Se species were determined by high per⁃formance liquid chromatography-inductively coupled plasma mass spectrometry(HPLC-ICP-MS).As for the identification of unkown Se compounds,the enzymatic hydrolysate was firstly con⁃centrated by removing impurities with a 3 kDa ultrafiltration centrifuge tube,then freeze-dried,and finally suspended in the initial mobile phase.A Kinetex F5 column(100 mm×2.1 mm,2.6µm)was selected for gradient elution.Mobile phases A and B were water and acetonitrile both prepared with 0.1%(volume fraction)formic acid.The unknown Se compounds were screened and identified by ultrahigh performance liquid chromatography-electrospray ionization high resolution tandem mass spectrometry(UPLC-ESI-Triple TOF MS)using unique isotope patterns,TOF MS and TOF MS/MS.The results showed that five Se species in C.Violifolia were determined by HPLC-ICP-MS,namely selenocysteine(SeCys_(2)),methylselenocysteine(MeSeCys),selenome⁃thionine(SeMet),selenite[Se(Ⅳ)]and selenate[Se(Ⅵ)].15 other Se compounds,including 7 reported and 8 unreported Se compounds,were identified by UPLC-ESI-Triple TOF MS.The structural formula of three unreported Se compounds,C_(4)H_(7)NO_(2)Se,C_(5)H_(8)O_(2)Se and C_(7)H_(9)NO_(2)Se were proposed.

关 键 词：富硒堇叶碎米荠 硒化合物 HPLC-ICP-MS UPLC-ESI-Triple TOF MS 鉴定 

分 类 号：O657.63[理学—分析化学] TS207.3[理学—化学]