Identification of Metabolites of Dendrobine in Human Liver Microsomes by UPLC/LTQ-Orbitrap-MS

LU Yan-liu; HUANG Si; WANG Min; QIU Min; WU Qin; LU Yuan-fu; HE Yu-qi

doi:10.13422/j.cnki.syfjx.2018040072

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Identification of Metabolites of Dendrobine in Human Liver Microsomes by UPLC/LTQ-Orbitrap-MS

更新时间：2024-01-04

- Identification of Metabolites of Dendrobine in Human Liver Microsomes by UPLC/LTQ-Orbitrap-MS
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 4, Pages: 72-77(2018)
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- DOI：10.13422/j.cnki.syfjx.2018040072
  CLC： R285
- Published：2018
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LU Yan-liu, HUANG Si, WANG Min, et al. Identification of Metabolites of Dendrobine in Human Liver Microsomes by UPLC/LTQ-Orbitrap-MS[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(4): 72-77.
DOI：

LU Yan-liu, HUANG Si, WANG Min, et al. Identification of Metabolites of Dendrobine in Human Liver Microsomes by UPLC/LTQ-Orbitrap-MS[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(4): 72-77. DOI： 10.13422/j.cnki.syfjx.2018040072.

摘要

目的：通过UPLC/LTQ-Orbitrap-MS技术对石斛碱在人肝微粒体中的主要代谢产物进行鉴定与分析。方法：运用人肝微粒体体外温孵法，模拟细胞色素P450酶（CYP450）及尿苷二磷酸葡萄糖醛酸基转移酶（UGT）酶反应体系，采用Thermo Q-Exactive型四极杆-静电场轨道阱高分辨质谱系统分析，Hypersil Gold C18色谱柱（2.1 mm×150 mm，1.9 μm），流动相0.1%甲酸水溶液-乙腈梯度洗脱，流速0.3 mL·min-1，电喷雾离子源（ESI）正离子模式检测。结果：通过精确相对分子质量和二级碎片离子信息，鉴定了石斛碱在人肝微粒体中Ⅰ相代谢的4个主要代谢产物，依次标记为M-250 （[M+H]+m/z 250.180 1），M-262 （[M+H]+m/z 262.180 0），M-280 （[M+H]+m/z 280.190 6），M-296 （[M+H]+m/z 296.185 6），以及Ⅱ相代谢的1个主要代谢产物，标记为M-440 （[M]+m/z 440.227 8）。结论：所建立的UPLC/LTQ-Orbitrap-MS测定人肝微粒体中石斛碱及其代谢产物的方法灵敏、快捷，可应用于石斛碱在人肝微粒中的代谢研究，同时为石斛碱的药物代谢动力学研究提供了实验依据。

Abstract

Objective: To establish a method for structural identification of the metabolites of dendrobine in the human liver microsome incubation system for the first time by UPLC/LTQ-Orbitrap-MS. Method: UPLC/LTQ-Orbitrap-MS was applied to systemically analyze the dendrobine metabolism in the liver microsomes of human.Chromatography separation was performed on a Thermo Hypersil Gold C18 column (2.1 mm×150 mm

1.9 μm) with acetonitrile-0.1% formic acid aqueous solution as mobile phase in gradient elution.Electrospray ionization (ESI) was applied and operated in positive ion mode. Result: Four major metabolites in phase Ⅰ metabolism and one major metabolites in phase Ⅱ metabolism of dendrobine marked as M-250([M+H]+ m/z 250.180 1)

M-262([M+H]+ m/z 262.180 0)

M-280([M+H]+ m/z 280.190 6)

M-296([M+H]+ m/z 296.185 6) and M-440([M]+ m/z 440.227 8) were observed in the human liver microsome incubation system. Conclusion: This study introduces a sensitive and efficient analysis method for detecting dendrobine and its metabolites in human liver mirosomes

and it lays a foundation for the further analysis on the pharmacokinetic research of dendrobine.

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