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

关键词:  猪尿  牛尿  利尿剂  高效液相色谱-串联质谱

Key words:  pig urine  bovine urine  diuretic  HPLC-MS/MS

摘要:

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10 µg/L和20 µg/L；利尿剂的测定在20 µg/L～200 µg/L范围内线性关系良好，线性相关指数r＞0.99；猪尿在20 µg/L、50 µg/L和100 µg/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 µg/L、50 µg/L和100 µg/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%。

Abstract:

A high performance liquid chromatography-tandem mass spectrometry method (HPLC-MS/MS) method 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 column. Mass spectrometry condition: 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 coefficients r＞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.