液相色谱-串联质谱法测定猪尿液中地西泮及其7种代谢物残留

Determination of Diazepam and Its Metabolite Residues in Swine Urine by UPLC-MS/MS

为建立快速测定猪尿中地西泮及其7种代谢物残留的超高效液相色谱-串联质谱(UPLC-MS/MS)确证方法,本研究优化了样品净化的阳离子交换固相萃取柱(PCX柱)及其淋洗洗脱条件。猪尿液酸化后直接经PCX柱净化,依次用水、60%甲醇水溶液淋洗,最后用5%氨化甲醇洗脱;选用BEH C18色谱柱分离,UPLCMS/MS进行检测,以基质匹配标准曲线定量。结果显示:8种药物在0.3~20.0μg/L范围内具有良好的线性关系(R^(2)≥0.995),检出限和定量限分别为0.1μg/L和0.3μg/L;各药物在3个添加浓度下回收率为73.6%~95.3%,日内、日间相对标准偏差分别为2.9%~18.6%(n=6)和2.2%~12.6%(n=3)。结果表明,本方法操作简单、快速,灵敏度高、特异性好。

In order to establish an ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)to determine diazepam and its seven metabolite residues(temazepam,nimetazepam,nordiazepam,oxazepam,nitrazepam,7-aminonitrazepam and 7-aminonimetazepam)in swine urine,a cation exchange solid phase extraction column(PCX column)and its conditions of elution were optimized.Swine urine was directly purified by PCX column after being acidized,washed with water and 60%methanol water solution in turn,and then eluted with 5%ammoniated methanol;then the pre-treated urine sample was separated by BEH C18 column,detected by UPLC-MS/MS,and quantified by matrix-matched external standard curves.The results showed that a good linear relationship(R^(2)≥0.995)among the eight drugs was obtained within 0.3 to 20μg/L,the limits of detection and quantification were 0.1 and 0.3μg/L respectively;the mean recoveries varied from 73.6%to 95.3%at the three concentrations,the relative standard deviations of intra and inter day were 2.9%to 18.6%(n=6)and 2.2%to 12.6%(n=3),respectively.It was concluded that the proposed method was simple and rapid with high sensitivity and specificity.