使用咖啡因作为探针快速测定人体细胞色素P-450 CYP1A2活性。

Use of caffeine as a probe for rapid determination of cytochrome P-450 CYP1A2 activity in humans.

作者信息

Ou-Yang D S, Huang S L, Xie H G, Wang C Y, Zhou H H

机构信息

Pharmacogenetics Research Institute, Hu-nan Medical University, Changsha, China.

出版信息

Zhongguo Yao Li Xue Bao. 1998 Jan;19(1):44-6.

PMID:10375757

Abstract

AIM

To develop a rapid HPLC method for the determination of cytochrome P-450 CYP1A2 activity.

METHODS

A 300-microL plasma was prepared by extraction with 5-mL chloroform/isopropanol (9:1), and beta-hydroxyletheophylline was added as internal standard (IS). Samples were separated on an ODS column by a gradient elution system, of which mobile phase consisted of 0.05% acetic acid, acetonitrile, and methanol. The compounds of interest were monitored at 282 nm by UV detector.

RESULTS

No potential interfering peaks were found. Paraxanthine (17X), IS and caffeine (137X) were rapidly eluted with baseline resolution, and their retention time was less than 13 min. The detection limits of both 17X and 137X were 0.1 mumol.L-1. Linear relations ranged over 1-100 mumol.L-1 and 1-200 mumol.L-1 with correlation coefficient of 0.9999 and 0.9987, respectively, for 17X and 137X. The coefficients of variation were within 6% for 17X, and 10% for 137X. The average recoveries for both compounds were ranged from 96% to 108%.

CONCLUSION

This method is sensitive and rapid, and can be used for population studies of CYP1A2.

摘要

目的

建立一种快速高效液相色谱法测定细胞色素P - 450 CYP1A2活性。

方法

取300μL血浆，用5mL氯仿/异丙醇（9:1）萃取，加入β - 羟基乙茶碱作为内标（IS）。样品在ODS柱上通过梯度洗脱系统分离，流动相由0.05%乙酸、乙腈和甲醇组成。用紫外检测器在282nm处监测目标化合物。

结果

未发现潜在干扰峰。副黄嘌呤（17X）、内标和咖啡因（137X）快速洗脱，基线分离，保留时间小于13分钟。17X和137X的检测限均为0.1μmol·L⁻¹。17X和137X的线性关系范围分别为1 - 100μmol·L⁻¹和1 - 200μmol·L⁻¹，相关系数分别为0.9999和0.9987。17X的变异系数在6%以内，137X的变异系数在10%以内。两种化合物的平均回收率在96%至108%之间。