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目的:建立同时测定大鼠血浆中CYP450探针药物氯唑沙宗(CYP2E1)和咪达唑仑(CYP3A4)浓度的高效液相色谱法。方法:色谱柱为Inertsil-C18(250 mm×4.6 mm,5μm);流动相为乙腈-20 mmol.L-1乙酸铵-乙酸(45∶55∶0.1,v/v);流速为1 mL.min-1;紫外检测波长(λ)为210nm;柱温为40℃。结果:氯唑沙宗和咪达唑仑均在0.05~30μg.mL-1的范围内线性关系良好,最低检测限均为0.05μg.mL-1,低、中、高三个浓度的回收率分别为96.47%±4.35%、112.56%±4.05%、112.64%±4.28%;85.68%±2.81%、93.53%±3.70%、110.67%±5.54%。结论:本方法操作简便,灵敏度高,快速可靠,可同时用于血浆中CYP450亚型酶的探针药物氯唑沙宗和咪达唑仑的浓度的测定。
Objective: To establish a high performance liquid chromatography (HPLC) method for the simultaneous determination of CYP2E1 and CYP3A4 concentrations in rat plasma. METHODS: The column was Inertsil-C18 (250 mm × 4.6 mm, 5 μm). The mobile phase consisted of acetonitrile-20 mmol.L-1 ammonium acetate-acetic acid (45:55:0.1, v / v) and the flow rate was 1 mL. min-1; UV detection wavelength (λ) of 210nm; column temperature of 40 ℃. Results: Both chlorzoxazone and midazolam showed good linearity in the range of 0.05 ~ 30μg.mL-1 with the detection limits of 0.05μg.mL-1. The recoveries of low, medium and high concentrations were respectively Were 96.47% ± 4.35%, 112.56% ± 4.05%, 112.64% ± 4.28%; 85.68% ± 2.81%, 93.53% ± 3.70%, 110.67% ± 5.54%, respectively. Conclusion: The method is simple, sensitive, fast and reliable, and can be used for the determination of concentration of chlorzoxazone and midazolam in the plasma CYP450 subtype enzyme.