【摘要】 目的:建立同时测定细胞色素P450 (cytochrome P450,CYP450)亚型CYP1A2、CYP2C9和CYP2E1活性的Cocktail探针药物溶液的高效液相色谱(HPLC)检测方法。方法:分别选择茶碱、甲苯磺丁脲和氯唑沙宗作为CYP1A2、CYP2C9和CYP2E1的特异性探针药物,根据其理化特点配制成Cocktail组合探针溶液。使用HPLC梯度洗脱法同时测定3种探针药物的血药浓度,并对其方法学进行验证。结果:本研究建立的HPLC法可以同时测定大鼠血浆中3种探针药物的浓度,各探针药物的吸收峰分离完全,无杂峰干扰;线性关系良好,线性范围为0.02～50μg/mL,最低检测限为0.02μg/mL。日内、日间精密度均小于10%,回收率在87.74%～112.39%之间,稳定性高,室温放置、-20℃冷冻1周和3周的RSD均小于10%。将该方法应用于研究大鼠体内此3种酶的活性,证明其具有较好的灵敏度。结论:经方法学验证,此法可同时测定3种探针药物茶碱、甲苯磺丁脲和氯唑沙宗的血浆浓度。更多还原

【Abstract】 Objective:To establish an HPLC method for simultaneously determination of the metabolic activities of cytochrome P450(CYP450) isoforms CYP1 A2, CYP2 C9和CYP2 E1. Methods: Cocktail probe solution was prepared by mixing3 specific probe drugs of CYP1 A2,CYP2 C9 and CYP2 E1, namely theophylline,tolbutamide and chlorzoxazone respectively.An HPLC method for simultaneous determination of the above 3 probes was established and validated. Results: An HPLC method for simultaneous determination of the plasma concentrations of 3 probe drugs was established.The absorption peak of each probe was separated without interference of impurity peak.For each probe, the linear range was 0.02 ～50μg/mL, the minimum detection limit was 0.02μg/mL, the intraday and interday precisions were less than 10%, the recovery was between87.74% ～112.39%, the RSD of stability after frozen 1 and 3 weeks was less than 10%. The method showed good sensitivity for pharmacokinetic study in vivo.Conclusion: The established HPLC method is efficient and reliable for simultaneous determination of theophylline, tolbutamide and chlorzoxazone, thus can be applied to activity evaluation of CYP1 A2, CYP2 C9 and CYP2 E1 with a cocktail approach.更多还原