高效液相色谱-串联质谱法测定动物尿液中8种利尿剂残留量

Determination of 8 Diuretics Residues in Animal Urine by HPLC-MS/MS

建立了动物尿液中氯噻嗪、氢氯噻嗪、氢氟噻嗪、氯噻酮、三氯噻嗪、甲氯噻嗪、呋噻米和依他尼酸等8种利尿剂残留检测的高效液相色谱-串联质谱（HPLC—MS／MS）方法。尿液样品经乙酸乙酯提取、阴离子交换固相萃取柱（PAX）净化、C18色谱柱分离、电喷雾离子化（ESI^-）和选择反应监测（SRM）方式采集，外标法定量。该方法的检测限和定量限分别为10μg／L和20μg/L；利尿剂的测定在20～200μg／L范围内线性关系良好，相关系数R^2大于0．99；猪尿在20、50和100μg／L三个添加浓度的平均回收率为74．0％～107．7％，批内RSD为5．2％～10．8％，批间RSD为5．7％～15．6％；牛尿在20、50和100μg／L三个添加浓度的平均回收率为71．1％～109．0％，批内RSD为3．9％～10．1％，批间RSD为6．4％～15．6％。

A high performance liquid chromatography- tandem mass spectrometry method （HPLC -MS/MS） has been established for the determination of 8 diuretics residues in pig urine and bovine urine. The samples of urine were extracted by ethyl acetate, cleaned through PAX cartridges and separated through a C18 colunm. Mass spectrometry condition was electrospray ionization （ESI^-） and selected reaction monitoring （SRM） mode. The quantification was carried out by the external standard method. The limit of detection （LOD） and quantification （LOQ） of the method were 10 μg/L and 20 μg/L respectively. The calibration curves have good linear within the range of 8 diuretics concentration from 20 μg/L to 200 μg/L, and the correlation coefficient was greater than 0.99. The average recoveries from spiked pig urine at three concentrations 20 μg/L, 50 μg/L and 100 μg/L ranged from 74.0% to 107.7%, and the within- and between- run relative standard deviations（RSDs） were 5.2% - 10.8% and 5.7% - 15.6% respectively. The average recoveries from spiked bovine urine at the same three concentrations ranged from 71.1% to 109.0%, and the within - and between - run RSDs were 3.9% - 10.1% and 6.4%-15.6% respectively.