Analysis of Metabolites and Metabolic Pathway of 2'-Methoxykurarinone in Rats

ZHAO Lin-lin; YU Ya-nan; HUANG Xiao-bin; WANG Huan; LIU Yi; CHEN Lei

doi:10.13422/j.cnki.syfjx.20180897

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Analysis of Metabolites and Metabolic Pathway of 2'-Methoxykurarinone in Rats

更新时间：2024-01-04

- Analysis of Metabolites and Metabolic Pathway of 2'-Methoxykurarinone in Rats
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 8, Pages: 79-84(2018)
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- DOI：10.13422/j.cnki.syfjx.20180897
  CLC： R285.5
- Published：2018
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ZHAO Lin-lin, YU Ya-nan, HUANG Xiao-bin, et al. Analysis of Metabolites and Metabolic Pathway of 2'-Methoxykurarinone in Rats[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(8): 79-84.
DOI：

ZHAO Lin-lin, YU Ya-nan, HUANG Xiao-bin, et al. Analysis of Metabolites and Metabolic Pathway of 2'-Methoxykurarinone in Rats[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(8): 79-84. DOI： 10.13422/j.cnki.syfjx.20180897.

摘要

目的：通过超高效液相色谱与质谱联用技术（UPLC-Q-Exactive Orbitrap-MS）分析2'-甲氧基苦参酮在大鼠体内的代谢产物和代谢途径。方法：按剂量50 mg·kg-1灌胃给予SD大鼠2'-甲氧基苦参酮后，由眼眦静脉丛取血并获得血浆。血浆样品经固相萃取柱处理后，以乙腈和0.1%甲酸水溶液为流动相，经Hypersil GOLD C18色谱柱分离后进入质谱，电喷雾离子源在负离子模式下进行一级和二级数据的采集，电喷射电压3 kV，离子传输管温度320℃，m/z 100~1 000，使用Xcalibur 4.0软件对数据进行处理，对比给药血浆与空白血浆的总离子流图，通过色谱和质谱信息分析，筛选出2'-甲氧基苦参酮在大鼠体内可能的代谢产物。结果：共检测到11个代谢产物，其中7个为Ⅰ相代谢产物，包括脱氢、羟基化、水合反应、羟基化和脱氢相结合的代谢物；其余4个为Ⅱ相代谢产物，包括葡萄糖醛酸化、硫酸化、葡萄糖醛酸化与羟基化相结合的代谢物。结论：UPLC-Q-Exactive Orbitrap-MS结合Xcalibur 4.0软件的方法能够提供分析物的结构信息，为鉴定2'-甲氧基苦参酮的代谢产物提供了可靠的检测手段；由检测到的代谢产物，推测2'-甲氧基苦参酮Ⅰ相代谢反应位置主要是异戊烯基基团。

Abstract

Objective: To profile and identify metabolites of 2'-methoxykurarinone in rat plasma by UPLC-Q-Exactive Orbitrap-MS. Method: SD rats were orally administered 2'-methoxykurarinone and blood samples were collected through retro orbital sinus to obtain plasma.The extraction of analytes from rat plasma were achieved by solid-phase extraction method on a HyperSep Retain PEP column.The mobile phase consisted of acetonitrile and 0.1% formic acid in water.Separation was achieved on a Hypersil GOLD column.The data acquisition was carried out by electrospray ionization (ESI) with full scan and second stage mass scan in negative ion mode (spray voltage of 3 kV

capillary temperture at 320℃

m/z 100-1 000).Possible metabolites were detected and identified according to the information of chromatograph and mass spectrometry. Result: Seven phase Ⅰ metabolites and four phase Ⅱ metabolites of 2'-methoxykurarinone were detected in rat plasma.These metabolites were formed by a series of metabolic reactions

including dehydrogenation

hydroxylation

hydroxylation+dehydrogenation

hydration

sulfation conjugation

glucuronidation

and hydroxylation+glucuronidation. Conclusion: The method of UPLC-Q-Exactive Orbitrap-MS coupled with Xcalibur 4.0 can provide structural information of analytes

which provides a reliable method for the identification of metabolites of 2'-methoxykurarinone.The phase Ⅰ metabolic reaction of 2'-methoxykurarinone occurs mainly in the prenyl moiety.

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