反相高效液相色谱法同时测定人血浆中对乙酰氨基酚/曲马朵及其代谢物的含量

目的:建立一种用高效液相色谱法(HPLC)同时测定人血浆中对乙酰氨基酚/曲马朵及其代谢物氧去甲基曲马朵(M1)浓度的方法。方法:以间乙酰氨基酚为内标,血浆样品经碱化后,用乙酸乙酯提取。HPLC-UV测定采用DiamonsilTM C18色谱柱(250mm×4.6mm),流动相为0.01mol·L-1磷酸二氢钾缓冲液(含三乙胺0.005mol·L-1,磷酸调pH3.0)-乙腈(79:21),流速:1.0ml·min-1,紫外检测波长:218nm。结果:本实验条件下各物质分离良好,对乙酰氨基酚在0.2-21.6μg·ml-1范围呈线性,曲马朵及其代谢物M1分别在10-720和5-160ng·ml-1范围呈线性。对乙酰氨基酚的最低检测限为0.2μg·ml-1,曲马朵和M1的最低检测限分别为10和5ng·ml-1。三种物质的方法回收率均在80%-120%范围内,提取回收率均在75%以上。结论:本法简单、快速,可用于人血浆中对乙酰氨基酚/曲马朵及M1的同时检测。

SIMULTANEOUS DETERMINATION OF PARACETAMOL/TRAMADOL AND ITS METABOLITE IN HUMAN PLASMA BY REVERSED-PHASE HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Objective:To establish a reversed-phase high performance liquid chromatography(HPLC) method for the determination of paracetamol/tramadol and its metabolite O-desmethyltramadol(M1)in human plasma. Methods: M-acetaminphen was used as an internal standard,and alkalinized samples of plasma were extracted with ethyl acetate. Chromatographic separation was achieved with a DiamonsilTM C18 analytical column(250 mm ×4.6 mm). The mobile phase was 0.01 mol·L-1 potassium dihydrogen phosphate (0.005 mol·L-1 triethylamine,pH 3.0 adjusted by phosphoric acid)-acetonitrile(79∶21),and flow rate was 1.0 ml·min-1.The drug concentrations were measured by UV at 218 nm. Results:The calibration curves were linear in the concentration range of 0.2-21.6 μg·ml-1, 10-720 ng·ml-1 and 5-160 ng·ml-1 for paracetamol,tramadol and M1, respectively. The limit of quantition(LOQ) was 0.2 μg·ml-1for paracetamol,10 ng·ml-1 for tramadol and 5 ng·ml-1 for M1. The recovery of the three samples was within 80%-120%, the absolute recovery was above 75%. Conclusion:This method is simple and rapid,especially for quantitative analysis of paracetamol/tramadol in human plasma simultaneously.