HPLC测定人血浆中盐酸美沙酮的浓度

Determination of Plasma Concentrations of Methadone Hydrochloride by HPLC

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 develop an HPLC method for the determination of methadone hydrochloride in plasma.METHODS Agilent ZORBAX SB-C18（4.6 mm×250 mm,5 μm） column was used.The mobile phase was used acetonitrile-phosphate（pH 2.5）=32∶68.The flow rate was 1.5 mL·min.1 and the UV detection wavelength was 206 nm.The internal standard was trihexyphenidyl hydrochloride.The plasma sample was extracted with ether,the organic phase was then extracted twice with 100 μL hydrochloric acid,took 20 μL sample from water-phase for analysis.RESULTS The concentration of methadone hydrochloride had a good linear relationshipin within 0.1.10 mg·L.1（r=0.999 6）;the lower limit of quantification was 0.05 mg·L.1;the mean relative recovery was（96.89±0.64）%;the average absolute recovery was（77.68±2.47）%.The within-day and the between-day RSD were less than 5%.CONCLUSION The method is simple,rapid,accurate,and can be used for drug determination of methadone hydrochloride in plasma.