安非拉酮体内外代谢研究
In vitro and in vivo metabolism of amfepramone
王鑫 1杨欢 1赵君博 1向平1
作者信息
- 1. 司法鉴定科学研究院,上海市法医学重点实验室,上海市司法鉴定专业技术服务平台,司法部司法鉴定重点实验室,上海 200063
- 折叠
摘要
目的 通过液相色谱-高分辨质谱(LC-HRMS)检测分析大鼠肝微粒体模型中安非拉酮的代谢物并与安非拉酮服用者的真实尿液进行对比,从而对体外大鼠肝微粒体模型预测体内代谢物的一致性进行评价研究.方法 向Wistar大鼠肝微粒体中加入安非拉酮标准品5 μg,模拟人体内的代谢过程孵育 1 h,真实尿液经乙腈沉淀蛋白后,利用液相色谱Q Exactive组合型四极杆Orbitrap质谱仪(LC-Q-Exactive-Orbitrap-MS)分析体内外代谢物以查明代谢途径和体内生物标志物.结果 安非拉酮在大鼠肝微粒体体外模型中共产生了 10 种I相代谢产物,其中 8 种I相代谢产物在安非拉酮服用者的真实尿液样本中均有发现.推断主要代谢途径为羰基还原、N-脱烷基化、羟基化反应等.该体外代谢模型试验方法简便快捷,可在一定程度上预测安非拉酮体内代谢产物.安非拉酮一些代谢物如N-脱烷基化代谢物M2(乙卡西酮)和M4(卡西酮)等来源不唯一,因此不能单独作为安非拉酮摄入的特征生物标志物.结论 本研究发现安非拉酮的代谢存在羟基化途径,真实尿液中发现4 个羟基化代谢物.初步阐明了安非拉酮体内外代谢物、代谢位点和代谢途径,以为后续研究安非拉酮药代动力学、法医学评价体系的构建提供理论依据.
Abstract
Objective To detect and analyze the in vitro metabolites of amfepramone using a rat liver microsomal model by liquid chromatography coupled with high-resolution tandem mass spectrometry(LC-HR-MS/MS),and compare them with those derived from authentic urine samples collected from amfepramone users,so as to evaluate the consistency of the in vitro rat liver microsomal model in predicting the metabolites in vivo.Methods Wistar rat liver microsomes were exposed to 5 μg amfepramone and subsequently incubated for 1 h to simulate the metabolic process in the human body.The authentic urine samples collected from amfepramone users were precipitated with acetonitrile.Results A total of of ten phase I metabolites of amfepramone were produced in the in vitro rat liver microsome model.Eight out of the ten in vitro phase I metabolites were also found in authentic urine samples collected from amfepramone users.The primary metabolic pathways were deduced to be carbonyl reduction,N-dealkylation,and hydroxylation.This in vitro metabolic model-based approach proved to be a simple and rapid method for predicting amfepramone metabolites in vivo.However,it was observed that certain metabolites of amfepramone,including M2(ethcathinone)and M4(cathinone),possess non-exclusive origins,limiting their suitability as standalone biomarkers for detecting amfepramone use in urine specimens.Conclusion This study revealed the involvement of the hydroxylation pathway in amfepramone metabolism.Additionally,four new metabolites were identified in real urine samples from amfepramone users.These preliminary results contribute valuable insights into the pharmacokinetics and forensic analysis of amfepramone,providing a theoretical foundation for further research in these domains.
关键词
法医毒物分析/安非拉酮/大鼠肝微粒体/代谢途径/液相色谱-高分辨质谱(LC-HRMS)Key words
Forensic toxicology/Amfepramone/Liver microsomes/Metabolic pathway/LC-HRMS引用本文复制引用
出版年
2024
NETL
NSTL
万方数据