分子印迹固相萃取-高效液相色谱法测定尿液中甲基苯丙胺和苯丙胺

Simultaneous determination of methamphetamine and amphetamine in urine by high-performance liquid chromatography with molecularly imprinted solid-phase extraction

目的:建立快速、准确的尿液中甲基苯丙胺(MA)及其代谢产物苯丙胺(AMP)的分子印迹固相萃取(MISPE)高效液相色谱测定方法。方法:尿液与10 mmol/L醋酸铵缓冲液(pH 8.0)混合后离心分离(8000 rpm),采用预先活化过的分子印迹固相萃取柱净化,以醋酸/乙腈溶液洗脱,在XBridge RP18色谱柱上,以甲醇-乙腈-水溶液(15∶30∶55,V/V/V)为流动相,采用210 nm波长进行检测。结果:MA和AMP的加标回收率在88.5%～97.0%范围,其相对标准偏差均小于5.0%,在0.05 mg/L～15.0 mg/L范围呈现良好的线性,其回归系数大于0.999,检出限(LODs)分别为0.005 mg/L和0.01 mg/L。结论:本方法回收率高,净化效果显著,稳定性好,杂质干扰少,可满足临床吸毒病人尿液中MA和AMP的检测要求。

Objective：To develop a method for the simultaneous determination of methamphetamine（MA） and amphetamine（AMP） in urine by high-performance liquid chromatography with molecularly imprinted solid-phase extraction（MISPE）.Methods：After the urine samples were mixed with 10 mmol/L ammonium acetate buffer（pH8.0） and centrifuged at 8000 rpm,the clean up procedure was optimized on an activating molecularly imprinted solid-phase extraction cartridge and the elution was carried by acetate acid/ acetonitrile.The separation was performed on an XBridge RP18 column by a mobile phase consisting of methanol-acetonitrile-water（15：30：55,V/V/V）.Detection was carried out by an ultraviolet detector at 210 nm.Results：Calibration curves of MA and AMP were linear within the range of 0.05 mg/L～15.0 mg/L with correlation coefficients of more than 0.999.The limits of detection（LODs） were 0.005 mg/L and 0.01 mg/L for MA and AMP,respectively.The extraction recoveries were between 88.5%～97.0%,and the RSDs were less than 5.0%.Conclusion：This developed method is simple,sensitive and accurate.It is suitable for the detection of MA and AMP residues in biological specimens for the clinical diagnosis of drug abuse patients.