HPLC－电化学检测法检测血浆中的盐酸纳洛酮和盐酸纳曲酮

Determination of Naloxone Hydrochloride and Naltrexone Hydrochloride in Plasma by HPLC with Electrochemical Detector

本文建立了简便、灵敏的定量分析血浆中盐酸纳洛酮和盐酸纳曲酮的高效液相色谱－电化学检测法。血浆样品用苯－乙醇（９∶１）混合溶剂提取，在Ｃ１８柱上以０．１ｍｏｌ／Ｌ磷酸二氢钾－甲醇（８４∶１６）为流动相进行分离，用电化学检测器在７６０ｍＶ的工作电压下进行检测，色谱峰分离完全，血浆本底无干扰。将盐酸纳曲酮和盐酸纳洛酮互为内标，测得两药在２．５～１６０ｎｇ／ｍｌ的范围内具有良好的线性关系，血浆中盐酸纳洛酮和盐酸纳曲酮的回收率分别为７６．８％～８９．６％和７８．６％～８４．６％，日内ＲＳＤ＜４．８％，日间ＲＳＤ＜９．１％，最低检出浓度为１ｎｇ／ｍｌ血浆。应用本法对犬ｉｖ盐酸纳洛酮和盐酸纳曲酮的药代动力学进行了研究，报告了相应的动力学参数。

A simple and sensitive HPLC-electrochemical detection method has been developed for the determination of naloxone hydrochloride and naltrexone hydrochloride in plasma. Using a mixed solvent of benzene-ethanol (9∶1) the two drugs were extracted from plasma, separated on a C18 column with the mobile phase consisted of 0.1mol/L KH2PO buffer-methanol (84∶ 16) and assayed by electrochemical detection at an oxidation potential of 760 mV. The peaks were separated well without interference from the plasma. In the method naloxone hydrochloride and naltrexone hydrochloride were used as internal standard for each other. The linear ranges of the calibration curves for the two drugs were 2.5～160ng/ml and the minimal detectable concentration was 1.0ng/ml. The recoveries of naloxone hydrochloride and naltrexone hydrochloride were found to be 76.8%～89.6% and 78.6%～84.6% respectively. The RSD wrthin-day and day-to-day were less than 4.8% and 9.1% for the two drugs. The method has been used in pharmacokinetic studies of naloxone hydrochloride or naltrexone hydrochloride in dog after iv 5.0mg and their pharmacokinetic parameters were reported.