超高效液相色谱串联质谱法测定尿液中草铵膦及3种代谢物浓度
Determination of glufosinate ammonium and three metabolites in urine by ultra performance liquid chromatography-tandem mass spectrometry
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摘要:
背景 目前对尿液中草铵膦及3种代谢物的检验尚未形成统一标准，在一定程度上影响职业暴露风险的准确评估。建立快速有效的检验方法，对保障职业安全和公众健康具有重要意义。
目的 建立一种超高效液相色谱串联质谱法，用于同时检测尿液中草铵膦及3种代谢物。
方法 比较稀释溶剂以及稀释倍数对草铵膦及3种代谢物响应值的影响，分析不同固相萃取柱对目标物的保留能力，以及不同色谱柱、流动相体系对色谱峰的影响。采用基质效应匹配外标法定量。以加标回收率评估方法的准确度，批内和批间相对标准偏差评价方法的精密度，并采集30名正常人群的尿液样本进行方法的应用考察。
结果 本研究采用0.2 mL水、0.6 mL乙腈稀释尿液样本，通过HLB固相萃取柱净化后，以Dikma Polyamino HILIC色谱柱分离，0.5 mmol·L−1乙酸铵及0.1%氨水的水-乙腈体系为流动相进行梯度洗脱，可以获得良好的峰形和质谱响应。4种目标物在0.5~50 ng·mL−1浓度范围内线性关系良好，相关系数
r 均大于0.998，检出限为0.56~2.86 μg·L−1，定量限为1.87~29.54 μg·L−1，加标回收率范围为75.0%~103.6%，批内及批间精密度范围分别为2.5%~8.1%和4.3%~9.3%。应用本法对30份受试者尿液样本进行检测，未检测出目标物。结论 方法操作简单、快速，灵敏度及准确度高，并且无需衍生化，适用于人体尿液中草铵膦及其代谢物的测定。
Abstract:Backgroud At present, there is no unified standard for the detection of glufosinate ammonium and three metabolites in urine, which affects the accurate assessment of occupational exposure risk to a certain extent. It is of great significance to establish a rapid and effective inspection method to ensure occupational safety and public health.
Objective To establish an ultra performance liquid chromatography-tandem mass spectrometry for simultaneous determination of glufosinate ammonium and three metabolites in urine.
Methods The effects of dilution solvents and dilution ratios on the response values of glufosinate ammonium and three metabolites were compared, and the retention capacities of solid phase extraction columns for targets as well as the effects of chromatographic columns and mobile phase systems on chromatographic peaks were analyzed. Samples were quantified by matrix effect matching external standard method. Accuracy of the method was evaluated by recovery rate of standard addition, and precision of the method was evaluated by relative standard deviation of intra-day and inter-day measurements. Urine samples of 30 health individuals were collected to evaluate the application of the method.
Results The urine samples were diluted with 0.2 mL water and 0.6 mL acetonitrile, purified by HLB solid phase extraction columns, and separated by Dikma Polyamino HILIC columns, and gradient elution was carried out with 0.5 mmol·L−1 ammonium acetate and 0.1% ammonia water as mobile phase, which achieved a good peak shape and mass spectrum response. The linearities of the four target compounds were good in the range of 0.5-50 ng·mL−1, and the correlation coefficients (
r ) were all greater than 0.998. The detection limits were 0.56-2.86 μg·L−1, the quantification limits were 1.87-29.54 μg·L−1, and the recovery rates of standard addition ranged from 75.0% to 103.6%, The relative standard deviations of intra-batch and inter-batch were from 2.5% to 8.1% and from 4.3% to 9.3% respectively. The method was applied to detect 30 urine samples of subjects, and no target was detected.Conclusion The method is simple, rapid, sensitive, and accurate. It is suitable for the determination of glufosinate ammonium and its metabolites in human urine without derivatization.
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