HPLC-MS/MS法测定汗液中美托洛尔及其代谢物α-羟化美托洛尔的质量浓度

Simultaneous determination of metoprolol and its metabolite α-hydro-metoprolol secreted in human sweat by HPLC-MS/MS

目的建立高效液相色谱-串联质谱法(HPLC-MS/MS)测定汗液中美托洛尔(metoprolol,MET)及其代谢产物α-羟化美托洛尔(α-hydro-metoprolol,α-MET)的质量浓度。方法采用Finnigan液相色谱仪串联LXQ离子阱质谱仪检测,流动相为乙腈-甲醇-体积分数0.1%甲酸水溶液(体积比20∶20∶60),采用SRM模式,正离子模式监测。采用内标法计算待测样品的质量浓度,内标物为盐酸普萘洛尔(proprandoIhydroc.hloride,PRO)。结果MET在载药量0.5~50ng内、α-MET在载药量2.5~50ng内线性关系良好;MET(α-MET)低、中、高三个质量浓度的回收率分别为89.9%(89.5%)、103.5%(83.6%)和92.5%(87.3%);汗液中MET检出时间为服药后1~18h,α-MET检出时间为服药后2~16h。结论本方法适用于同时测定汗液中美托洛尔及其代谢产物的含量,可为药物的体外监测提供参考。

Objective To establish a HPLC-MS/MS method for the determination of metoprolol and its metabolite ot-hydro-metoprolol secreted in human sweat. Methods The sweat sample was processed and chromatographed with the mobile phase consisting of acetonitrile-methanol - 0. 1% formic acid aqueous solution（20： 20： 60, V/V/V）. SRM mode was applied and operated in the positive ion mode. The drug concertation was calculated using internal standard method with propranolol hydrochloride as the internal standard substance. Results The standard curve of MET was linear over the drug loading range from 0. 5 - 50 ng. The standard curve of o^-MET was linear over the drug loading range from 2. 5 - 50 ng. The recovery of MET（ and a-MET） at low, medium and high concentrations were 89.9% （ 89.5% ）, 103.5% （ 83.6% ） and 92.5% （87.3%）, respectively. The MET in sweat could be detected from 1 h after administration through to 18 h,and the a-MET from 2 h through to 16 h. Conclusions This method applies for determining the concentration of metoprolol and its metabolite a-hydro-metoprolol secreted in human sweat, and is suit-able for clinical monitor.