非衍生化高效液相色谱-串联质谱法快速检测生物体液中草甘膦、草铵膦及代谢物

建立了一种非衍生化高效液相色谱-串联质谱快速检测生物体液中草甘膦、草铵膦及其代谢物等8种极性农药的方法。8种极性农药经Metrosep A Supp 5阴离子色谱柱(150 mm×4.0 mm,5μm)分离,以纯水-200 mmol/L碳酸氢铵溶液(含0.1%氨水)为流动相进行梯度洗脱,负离子多反应监测(MRM)模式进行检测。实验结果表明,8种极性农药在0.5~50 ng/mL范围内线性关系良好(r²>0.99),检出限(S/N≥3)为0.08~0.3 ng/mL,定量下限(S/N≥10)为0.3~1 ng/mL。方法的基质效应为86.5%~106%,目标化合物的回收率为81.5%~114%,日内相对标准偏差(RSD)为0.30%~2.8%,日间RSD为0.50%~5.3%。该方法无需复杂的衍生化过程,简便快速、灵敏度高、稳定性好,适用于生物体液中8种极性农药的检测。

Non-derivative Rapid Determination of Glyphosate,Glufosinate and Their Metabolites in Biological Fluids by High Performance Liquid Chromatography-Tandem Mass Spectrometry

Glyphosate and glufosinate are a class of carcinogenic,non-selective and internally suction herbicides,which have received more and more attention from the society.Much work about inspection or identification of glyphosate and glufosinate in the drug-related cases are required as poisoning cases of this two herbicides poisoning,suicide and other causes also occur from time to time.To develop a detection method for such herbicides in biological fluids,e.g.blood and urine could provide a technical support for the qualitative and detection of cases and provide a reference for clinical toxic analysis and treatment.Therefore,a non-derivative high performance liquid chromatography-tandem mass spectrometry was established for the rapid detection of 8 polar pesticides such as glyphosate and glufosinate and their metabolites in biological fluids.8 polar pesticides were separated on a Metrosep A Supp 5 anion column(150 mm×4.0 mm,5μm),with pure water-200 mmol/L ammonium hydrogen carbonate solution containing 0.1%ammonia water as mobile phase by gradient elution,and analyzed in negative ion multiple reaction monitoring(MRM)mode.In this study,the chromatographic and mass spectrometry conditions for the instrument were optimized,and the blood and urine of biological fluids were studied.The blood and urine samples were mixed with water and oscillated,then freeze-centrifugated with appropriate amount of acetonitrile for 10 min.The supernatant was in turn passed through a Cleanert PEP Plus solid phase extraction column and a 0.22μm organic micro-porous membrane,then collected for analysis.The experimental results showed that there were good linear relationships for 8 polar pesticides in the range of 0.5-50 ng/mL with their correlation coefficients(r²)larger than 0.99.The limits of detection(LOD,S/N≥3)and the limits of quantitation(LOQ,S/N≥10)of this method were in the ranges of 0.08-0.3 ng/mL and 0.3-1 ng/mL,respectively.The matrix effects of the method in blood and urine were in the range of 86.5%-106%,and the recoveries for the target compounds ranged from 81.5%to 114%,with intra-day relative standard deviations(intra-day RSD,n=6)and inter-day RSD of 0.30%-2.8%and 0.50%-5.3%,respectively.With the advantages of simple operation,time-saving and labor-saving,high sensitivity,good accuracy and good reproducibility,the developed method could be used for the rapid and accurate qualitative and quantitative analysis of 8 polar pesticides,to meet the testing and identification requirements of glyphosate,glufosinate-related toxic cases.