Simultaneous determination of 9 CYP450 enzymes in human liver microsomes using LC-MS/MS

Zhongbo SHI; Dezhang ZHAO; Junhao JIANG; Baogang XIE; Yan LI; Chao YU

doi:10.16016/j.1000-5404.202008230

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Simultaneous determination of 9 CYP450 enzymes in human liver microsomes using LC-MS/MS

Basic Medicine | 更新时间：2026-01-09

- Simultaneous determination of 9 CYP450 enzymes in human liver microsomes using LC-MS/MS
- Journal of Third Military Medical University Vol. 43, Issue 3, Pages: 211-217(2021)
- 作者机构：
  
  400016 重庆，重庆医科大学药学院
- 作者简介：
- 基金信息：
- DOI：10.16016/j.1000-5404.202008230
  CLC： R322.47;R345;R446
- Received：28 August 2020，
  
  Revised：2020-10-25，
  
  Published：15 February 2021
- 稿件说明：
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Zhongbo SHI, Dezhang ZHAO, Junhao JIANG, et al. Simultaneous determination of 9 CYP450 enzymes in human liver microsomes using LC-MS/MS[J]. Journal of Third Military Medical University, 2021, 43(3): 211-217.
DOI：

Zhongbo SHI, Dezhang ZHAO, Junhao JIANG, et al. Simultaneous determination of 9 CYP450 enzymes in human liver microsomes using LC-MS/MS[J]. Journal of Third Military Medical University, 2021, 43(3): 211-217. DOI： 10.16016/j.1000-5404.202008230.

摘要

目的

建立一种同时测定人肝微粒体中9种CYP450酶活性的LC-MS/MS方法(鸡尾酒法)，用于评估药物对CYP450酶活性的影响，以预判药物间发生相互作用的可能性。

方法

使用9种不同酶的底物作为探针与人肝微粒体共同孵育，硝苯地平作为内标物质。用Agilent 1290系列液相色谱仪进行洗脱，所有代谢产物均在多反应监测模式下进行分析，并对单探针法与鸡尾酒法测定的IC

值进行比较。

结果

该LC-MS/MS方法专属性好，色谱峰分离完全且无干扰。线性关系良好，9种代谢产物在标准曲线范围内的相关系数均＞0.99。方法准确度为85.34%~113.89%，日内RSD

12.49%，日间RSD

14.98%，符合相关标准。该鸡尾酒法测定的抑制剂的IC

值与单个探针底物的IC

值呈高度相关(

0.997 8)。

结论

该LC-MS/MS方法可同时对人肝微粒体中9种CYP450酶进行活性测定。

Abstract

Objective

To develop a liquid chromatography with tandem mass spectrometry (LC-MS/MS) to simultaneously detect the activities of 9 CYP450 enzymes in human liver microsomes in order to anticipate the possibility of drug-drug interactions.

Methods

Nine different enzyme substrates were used as probes to incubate with human liver microsomes

with nifedipine as internal standard. All metabolites were eluted by Agilent 1290 series liquid chromatograph and then analyzed under multiple reaction monitoring. The IC

values obtained by single substrate method and cocktail mixed substrate method were compared.

Results

Our established method had good specificity

and all the chromatographic peaks were completely separated without interference. The linear relationship was good. The correlation coefficients of 9 metabolites within the range of the standard curve were all greater than 0.99. The accuracy of the method was 85.34%~113.89%

the intra-day relative standard deviation (RSD) was

12.49%

and the inter-day RSD was

14.98%

which complied with the relevant standards. The IC

value of the inhibitor determined by this cocktail method was highly correlated with that of a single probe substrate (

0.997 8).

Conclusion

Our LC-MS/MS method can quickly and simultaneously measure the activities of 9 CYP450 enzymes in human liver microsomes.

关键词

Keywords

references

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