超高效液相色谱串联质谱法测定人尿中美沙酮  

Determination of methadone in human urine by ultra performance liquid chromatography tandem mass spectrometry

作　　者：和佳鸳 徐虹[1] 张春玲[1] 蒲彦利[1] 张俊[1] 

机构地区：[1]镇江市疾病预防控制中心,江苏镇江212000

出　　处：《中国卫生检验杂志》2017年第22期3215-3217,共3页Chinese Journal of Health Laboratory Technology

基　　金：江苏省临床医学科技专项(BL2014066)

摘　　要：目的建立快速测定人尿中美沙酮的超高效液相色谱串联质谱测定方法。方法尿样采用0.10 mol/L氢氧化钠溶液碱化后,经乙醚漩涡提取、离心,吸取上清液浓缩吹干,用80%甲醇定容后进液相色谱-质谱系统分析,同位素内标法定量。色谱柱采用Waters ACQUITY UPLC BEH C18柱(2.1 mm×100 mm,1.7μm),流动相为乙腈-0.1%甲酸,流速为0.4 ml/min,柱温为35℃,进行梯度洗脱;质谱采用电喷雾离子源,在正离子多反应监测模式(MRM)下进行检测。结果美沙酮在1 ng/ml^100 ng/ml时线性关系良好(r>0.999);检测限(LOD)和定量限(LOQ)分别为0.003 mg/L和0.01 mg/L;低、中、高3个浓度(2 ng/ml、10 ng/ml、50 ng/ml)的回收率分别为99.8%、100.1%、100.3%,精密度(RSD)分别为1.51%、2.72%、2.25%。结论该方法简便、灵敏、准确、稳定,适用于人尿中美沙酮的测定,可满足美沙酮临床研究尿样监测的要求。Objective To establish an ultra performance liquid chromatography tandem mass spectrometry（ UPLC-MS/MS）method for the determination of methadone in human urine. Methods The urine samples were extracted with diethyl ether by vortexing after alkalization with 0. 10 mol/L sodium hydroxide solution and then centrifuged. The extracts were concentrated to dry and then redissolved in 80% methanol,and finally analyzed by LC-MS/MS. The quantification was based on isotope internal standard method. The separation column was Waters ACQUITY UPLC BEH C18（ 2. 1 mm × 100 mm,1. 7 μm） at 35 ℃,and the mobile phase was consisting of acetonitrile-0. 1% formic acid with the flow rate of 0. 4 ml/min by gradient elution. The mass spectrometric detection was performed by electrospray ion source in positive ions multiple reaction monitoring mode（ MRM）.Results Good linearity was obtained when the concentration of the method was within 1 ng/ml-100 ng/ml（ r〉 0. 99）. Limits of detection（ LODs） and limits of quantification（ LOQs） were 0. 003 mg/L and 0. 01 mg/L,respectively. The recoveries of the method at low,medium and high concentration were 99. 8%,100. 1% and 100. 3%,respectively; and the RSDs were 1. 51%,2. 72% and 2. 25%,respectively. Conclusion The proposed method is simple,sensitive,accurate,and stable,which could be applied for the determination of methadone in human urine and meet the monitoring requirements for clinical research.

关 键 词：超高效液相串联质谱法 人尿 美沙酮 

分 类 号：R96[医药卫生—药理学]