手性固定相HPLC法测定大鼠血浆阿罗洛尔对映体及药代动力学研究

目的: 建立大鼠血浆中阿罗洛尔对映体的HPLC定量分析方法,并研究盐酸阿罗洛尔片在Wistar大鼠体内的手性药代动力学特征。方法: 血浆样品采用液-液萃取的方法进行前处理。采用Chiralpak AD-H(250 mm×4.6 mm, 5 μm)手性色谱柱,正己烷-乙醇-二乙胺-三乙胺(30:70:0.1:0.1)为流动相,流速0.4 mL·min^-1,检测波长315 nm。结果: d-阿罗洛尔和l-阿罗洛尔质量浓度均在0.025~1.25 μg·mL^-1范围内线性关系良好(r²>0.99)。阿罗洛尔对映体的回收率均大于94.1%,日内、日间RSD均小于15%。应用本文方法测定Wistar大鼠灌胃给药阿罗洛尔后的各对映体血浆浓度,获得相应的药动学参数,d-阿罗洛尔和l-阿罗洛尔的C_max、AUC_0-∞分别为0.225 μg·mL^-1、1.82 μg·h·mL^-1和0.226 μg·mL^-1、1.84 μg·h·mL^-1。结论: 该方法经方法学验证,可用于阿罗洛尔对映体在大鼠体内药代动力学研究,发现阿罗洛尔对映体在大鼠血浆药动学特征不存在立体选择性。

Objective: To establish a method for the determination of arotinolol enantiomers in rat plasma, and to study its pharmacokinetic features in Wistar rats. Methods: The plasma samples were pretreated with liquid-liquid extraction. The concentrations of arotinolol enantiomers in rat plasma were determined by HPLC with Chiralpak AD-H column (250 mm×4.6 mm, 5 μm) and UV detector. The mobile phase consisted of hexane-ethanol-diethylamine-triethylamine (30:70:0.1:0.1), with a flow rate of 0.4 mL·min^-1. The detection wavelength was set at 315 nm. Results: Good linear relationship was obtained in the range of 0.025-1.25 μg·mL^-1(r²>0.99). The recoveries of arotinolol enantiomers were above 94.1%. The intra-day and inter-day RSD for arotinolol were lower than 15%. After intragastric administration, the C_max and AUC_0-∞ of d-arotinolol were 0.225 μg·mL^-1 and 1.82 μg·h·mL^-1, respectively. The C_max and AUC_0-∞ of l-arotinolol were 0.226 μg·mL^-1 and 1.84 μg·h·mL^-1, respectively. Conclusion: The established method is proved suitable for the pharmacokinetic study of arotinolol in Wistar rats. In addition, our results indicate that arotinolol enantiomers have no chiral selectivity in Wistar rats.