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目的建立HPLC测定人血浆中盐酸美沙酮浓度的方法。方法采用Agilent ZORBAX SB-C18(4.6 mm×250 mm,5μm)色谱柱,以乙腈-磷酸盐缓冲液(pH 2.5)=32∶68为流动相,流速为1.5 mL.min-1,检测波长为206 nm,盐酸苯海索为内标,对血浆中的盐酸美沙酮进行检测。血浆样品以乙醚提取,取有机相用100μL盐酸二次萃取,取水相20μL进样分析。结果盐酸美沙酮浓度在0.1～10 mg.L-1内线性关系良好(r=0.999 6);定量下限为0.05 mg.L-1;平均相对回收率为(96.89±0.64)%;平均绝对回收率为(77.68±2.47)%;日内、日间RSD均<5%。结论该法简便、快速、准确,可用于血浆中盐酸美沙酮浓度的测定。
Objective To establish a HPLC method for the determination of methadone hydrochloride in human plasma. Methods The mobile phase was acetonitrile-phosphate buffer (pH 2.5) = 32:68 with a flow rate of 1.5 mL · min-1 on an Agilent ZORBAX SB-C18 column (4.6 mm × 250 mm, 5 μm) 206 nm, benehhydrochloride as an internal standard, the detection of methadone hydrochloride in plasma. The plasma samples were extracted with ether. The organic phase was extracted twice with 100 μL hydrochloric acid and the aqueous phase was extracted by 20 μL injection. Results The linear range of methadone hydrochloride was 0.1-10 mg.L-1 (r = 0.999 6). The lower limit of quantitation was 0.05 mg.L-1. The average relative recovery was (96.89 ± 0.64)%. The average absolute recovery (77.68 ± 2.47)%; day and day RSD were less than 5%. Conclusion The method is simple, rapid and accurate and can be used to determine the concentration of methadone hydrochloride in plasma.