Evaluation of Tetrahydropalmatine Enantiomers on the Activity of Five Cytochrome P450 Isozymes in Rats Using a Liquid Chromatography/Mass Spectrometric Method and a Cocktail Approach
Evaluation of Tetrahydropalmatine Enantiomers on the Activity of Five Cytochrome P450 Isozymes in Rats Using a Liquid Chromatography/Mass Spectrometric Method and a Cocktail Approach
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使用液相色谱/质谱法和混合法评价延胡索乙素对映体对大鼠五种细胞色素 P450 同工酶的活性
DOI：
10.1002/chir.22469
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发表时间：
2015-08-01
期刊：
影响因子：
2
通讯作者：
Hong, Zhanying
中科院分区：
文献类型：
--
作者：
Li, Wuhong;Zhao, Liang;Hong, Zhanying
目的是评价四氢巴马汀（THP）对映体在体内对5种细胞色素P450（CYP 450）同工酶活性的影响。建立了同时测定大鼠血浆中美托洛尔（2D 6）、咖啡因（1A 2）、氨苯砜（3A 4）、氯唑沙宗（2 E1）和甲苯磺丁脲（2C 9）5种特异性探针底物的液相色谱/质谱（LC-MS）法。以0.1%乙酸水溶液-乙腈为流动相，采用梯度洗脱，移动的进行分离。在同一次运行中，通过选择离子监测（SIM）的质谱检测在正离子模式（美托洛尔m/z 268，咖啡因m/z 195和氨苯砜m/z 249）和负离子模式（氯唑沙宗m/z 168和甲苯磺丁脲m/z 269）下操作。在0.050-25.0 μ g/mL的咖啡因和氨苯砜、0.025-10.0 μ g/mL的美托洛尔、0.050-50.0 μ g/mL的氯唑沙宗和0.25-100.0 μ g/mL的甲苯磺丁脲浓度范围内，线性关系良好（r（2）>0.99）。日内和日间精密度均小于12.09%。基质效应在87.50%~ 109.25%之间，绝对回收率大于70%。该方法已成功地应用于评价THP对映体对CYP 450同工酶活性的鸡尾酒法的影响。5种探针药物的药代动力学结果表明，THP对映体之间存在立体选择性差异，d-THP对CYP 2D 6和CYP 1A 2有抑制作用，而l-THP对CYP 1A 2有抑制作用，对CYP 3A 4和CYP 2C 9有诱导作用。Chirality 27：551-556，2015. (c)2015 Wiley Periodicals，Inc.
The aim was to evaluate the effects of tetrahydropalmatine (THP) enantiomers on the activity of five cytochrome P450 (CYP450) isozymes in vivo. A liquid chromatography / mass spectrometric (LC-MS) method was developed for simultaneous determination of five specific probe substrates including metoprolol (2D6), caffeine (1A2), dapsone (3A4), chlorzoxazone (2E1), and tolbutamide (2C9) in rat plasma. Analytes were separated with the mobile phase consisting of 0.1% acetic acid aqueous solution and acetonitrile in a gradient elution. The mass spectrometric detection via selected ion monitoring (SIM) was operated in both positive ion mode (for metoprolol m/z 268, caffeine m/z 195, and dapsone m/z 249) and negative ion mode (for chlorzoxazone m/z 168 and tolbutamide m/z 269) in the same run. Linear correlation was obtained (r(2)>0.99) over the concentration range of 0.050-25.0 mu g/mL for caffeine and dapsone, 0.025-10.0 mu g/mL for metoprolol, 0.050-50.0 mu g/mL for chlorzoxazone, and 0.25-100.0 mu g/mL for tolbutamide. Intra- and interday precision were less than 12.09%. The matrix effect ranged from 87.50% to 109.25% and the absolute recoveries were greater than 70%. The method was successfully applied to evaluate the effect of THP enantiomers on the activity of CYP450 isozymes by a cocktail approach. The pharmacokinetic results of five probe drugs indicated that there were stereoselective differences between the two THP enantiomers, i.e., d-THP had the potential to inhibit the activities of CYP2D6 and CYP1A2 isozymes, while l-THP inhibited CYP1A2 isozyme and induced CYP3A4 and CYP2C9 isozymes. Chirality 27:551-556, 2015. (c) 2015 Wiley Periodicals, Inc.