He Fan, Bi Hui-Chang, Xie Zhi-Yong, Zuo Zhong, Li Jian-Kang, Li Xin, Zhao Li-Zi, Chen Xiao, Huang Min
School of Pharmaceutical Sciences, Sun Yat-sen University, 74 Zhongshan Road II, Guangzhou 510080, China.
Rapid Commun Mass Spectrom. 2007;21(5):635-43. doi: 10.1002/rcm.2881.
A rapid liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed for the determination of six cytochrome P450 (CYP) probe substrate metabolites including paracetamol (PAR) for CYP1A2, 4-hydroxytolbutamide (OHTOL) for CYP2C9, 5-hydroxyomeprazole (OHOMe) for CYP2C19, dextrorphan (DEXM) for CYP2D6, 6-hydroxychlorzoxazone (OHCHL) for CYP2E1 and dehydronifedipine (DNIF) for CYP3A4. The triple-quadrupole mass spectrometer was operated in both positive and negative modes, and selective reaction monitoring was used for quantification. The method was validated over the concentration ranges (0.075/0.04/0.05/0.02/0.1/0.0625 microM to 4.8/2.56/3.2/1.28/6.4/4.0 microM) for PAR/OHTOL/OHOME/DEXP/OHCHL/DNIF analytes with acceptable accuracy and precision. The inhibitory effect on the six CYP enzymes has been verified with their known specific inhibitors. This high-throughput inhibition screening approach has been successfully applied to study the inhibitory effects of 18 terpenoids on CYP enzymes. Among them, tanshinone IIA and cryptotanshinone are found to be potent inhibitors to CYP1A2, while artemisinin is a marginal inhibitor to CYP1A2 and glycyrrhetic acid is a weak inhibitor to CYP2C9.
建立了一种快速液相色谱/串联质谱（LC/MS/MS）方法，用于测定六种细胞色素P450（CYP）探针底物代谢物，包括用于CYP1A2的对乙酰氨基酚（PAR）、用于CYP2C9的4-羟基甲苯磺丁脲（OHTOL）、用于CYP2C19的5-羟基奥美拉唑（OHOMe）、用于CYP2D6的右美沙芬（DEXM）、用于CYP2E1的6-羟基氯唑沙宗（OHCHL）以及用于CYP3A4的脱氢硝苯地平（DNIF）。三重四极杆质谱仪以正、负两种模式运行，并采用选择性反应监测进行定量分析。该方法在PAR/OHTOL/OHOME/DEXP/OHCHL/DNIF分析物的浓度范围（0.075/0.04/0.05/0.02/0.1/0.0625 microM至4.8/2.56/3.2/1.28/6.4/4.0 microM）内进行了验证，具有可接受的准确度和精密度。用已知的特异性抑制剂验证了对六种CYP酶的抑制作用。这种高通量抑制筛选方法已成功应用于研究18种萜类化合物对CYP酶的抑制作用。其中，丹参酮IIA和隐丹参酮被发现是CYP1A2的强效抑制剂，而青蒿素是CYP1A2的边缘抑制剂，甘草次酸是CYP2C9的弱抑制剂。
