内源性甾体激素GC/MS分析中酮基衍生规律的研究  

Study on the Keto Groups Derivatization of Endogenous Steroid Hormones with GC/MS

作　　者：李慧 付玉娟[1] 焦丽丽 刘淑莹 宋凤瑞 吴巍[1] LI Hui;FU Yu-juan;JIAO Li-li;LIU Shu-ying;SONG Feng-rui;WU Wei(Jilin Ginseng Academy,Changchun University of Chinese Medicine,Changchun 130117,China;Changchun Institute of Applied Chemistry Chinese Academy of Sciences,Changchun 130022,China)

机构地区：[1]长春中医药大学,吉林省人参科学研究院,吉林长春130117 [2]中国科学院长春应用化学研究所,吉林长春130022

出　　处：《质谱学报》2019年第6期499-509,共11页Journal of Chinese Mass Spectrometry Society

基　　金：国家自然科学基金项目(81873193);吉林省科技发展计划项目(20180520062JH);吉林省中医药科技项目(2018DZ01);吉林省教育厅项目(JJKH20190473KJ)资助

摘　　要：本实验以甲氧胺(MOX)和N-甲基-N-(三甲基硅烷)三氟乙酰胺(MSTFA)为衍生化试剂,采用肟化-硅烷化方法对12种内源性甾体激素进行衍生,通过GC/MS解析鉴定衍生产物结构,研究甾体激素的衍生规律。结果表明,肟化-硅烷化反应可以提高甾体激素的信号,但孕酮(P)、雄烯二酮(ASD)、雌酮(E1)、睾酮(T)、皮质甾酮(B)、可的松(E)受共轭结构影响,产生多种同分异构体。[M-31]^+和[M-15]^+,[M-90]^+和[M-72]^+可分别作为肟化和硅烷化反应的鉴定依据。在80℃,加热40 min,酮基与MOX的摩尔比为1∶20是肟化反应的最佳条件。采用单反应离子检测(SRM)模式,衍生后甾体激素的检测灵敏度可达1μg/L,线性范围为1~200μg/L。该方法可为相关激素类成分的分析提供检测技术支持。Steroid hormones are a kind of aliphatic hydrocarbon compounds.They are synthesized in central or peripheral nervous system which are considered as potential biomarkers.High sensitivity method was required for steroid hormones detection for their poor responses and low physiological concentrations.Gas chromatography-mass spectrometry(GC/MS)combined with pre-column derivatization is commonly used in detection of compounds such as steroid hormones.In this work,derivation of oximation conditions were studied based on 12 steroid hormones.Oximation-silylation method was applied for the derivatization of dehydroeppiandrosterone(DHEA),dihydrotestosterone(DHT),androsterone(ADL),epiandrosterone(Epian),pregnenolone(Preg),testosterone(T),estrone(E1),androstenedione(ASD),corticosterone(B),cortisone(E),progesterone(P)and allopregnanolone(AP).Twelve endogenous steroid hormones were processed with methoxyamine(MOX)and N-methyl-N-(trimethylsilane)trifluoroacetamide(MSTFA).Oximation-silanization reactions were carried out on ketone group and hydroxyl group separately which could avoid enolate reaction of ketone group.When processed with MOX,characteristic peaks of[M-31]^+and[M-15]^+were observed in the electron bombardment(electron-impact,EI)mass spectra,mainly due to the loss of[—OCH3]and[—CH3]groups.Therefore peaks of[M-31]^+and[M-15]^+could be considered as the identification for ketoximation.At the same time,characteristic peaks of[M-73]^+,[M-90]^+due to the loss of[—Si(CH3)3]or[—O—Si(CH3)3]groups from[M]^+were observed when steroid hormones were further silylated with MSTFA.Peaks of[M-73]^+and[M-90]^+could be considered as the basis for hydroxysilylation.Preg,DHEA,AP,ADL,Epian,DHT generated single derivatization with good peak shape.For the existence of conjugated groups in P,ASD and T structures,isomer products generated after derivatization.The ion peak shape was poor and noise was high.For B,and E six derivatives were observed in each derivatives after reaction.Then oximation conditions for endogenous steroid ho

关 键 词：气相色谱-质谱法(GC/MS) 甾体激素 酮基 衍生 

分 类 号：O657.63[理学—分析化学]