超高效液相色谱-串联质谱同时检测血液中29种农药

Detection of 29 pesticides in the blood by ultra high performance liquid chromatography-series mass spectrometry

本文采用超高效液相色谱-串联质谱法(UPLC-MS/MS)和固相萃取法(SPE)建立了血液中29种农药同时筛查、定性、定量分析的方法,血液经4%磷酸水溶液稀释后,震荡10min,以8000r·min-1转速离心10min,取上清液过3mL甲醇和3mL水活化好的Oasis Prime HLB(3cc,60mg)固相萃取小柱,使用3mL5%甲醇水淋洗,3mL乙腈甲醇混合溶剂(90:10)洗脱,接收洗脱液后在40℃条件下氮吹仪吹干,使用0.5mL初始流动相复溶,震荡10s后,过0.22μm水膜,装液质小瓶后进样分析。采用ACQUITY UPLC HSS C18色谱柱(150 mm×2.1mm,1.8μm)分离,流动相为0.1%甲酸乙腈-水/甲酸/甲酸铵(5mmol,pH=3),梯度洗脱,电喷雾电离正离子模式(ESI+),多反应选择离子监测模式(MRM)检测。29种农药的检出限为0.1 ng·mL^-1~5 ng·mL^-1,定量限为0.5 ng·mL^-1~10 ng·mL^-1,回收率为62.4%~97.4%,基质效应为82.8%~109%,相对标准偏差小于10.3%,相关系数均大于0.99,线性关系良好范围为10 ng·mL^-1~1000ng·mL^-1。本文方法灵敏度高,可以对血液中29种农药成分进行筛查、定性、定量分析,能够满足实际血液样品中农药成分检测的需求。

In this paper,ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)and solid phase extraction(SPE)were used to establish a method for simultaneous screening,qualitative and quantitative analysis of 29 pesticides in blood.After the blood was diluted with 4%phosphoric acid solution,shaken for 10 min,centrifuged at 8000 r·min-1 for 10 min.Oasis Prime HLB(3 cc,60 mg)solid phase extraction cartridge was activated with 3 mL methanol and 3 mL water,then the supernatant was taken through a solid phase extraction cartridge.Rinse the solid phase extraction cartridge with 3 mL 5%methanol water,and elute the target compound using 3 mL mixed solvent of acetonitrile and methanol(90∶10).After receiving the eluent,it was dried by nitrogen at 40°C,reconstituted with 0.5 mL initial mobile phase,shaken for 10 s,and passed through a 0.22μm water film.The filtrate was loaded into a vial dedicated to liquid chromatography and mass spectrometry,and the sample was analyzed.The target compound was separated using an ACQUITY UPLC HSS C18 column(150 mm×2.1 mm,1.8μm).The mobile phase used 0.1%formic acid acetonitrile-water/formic acid/ammonium formate(5 mmol,pH=3).Set the mobile phase gradient to elute and separate the target compound.Electrospray ionization,positive ion mode(ESI+),and monitor target compounds in multiple reaction selective ion monitoring mode(MRM).The detection limit of 29 pesticides was 0.1 ng·mL^-1~5 ng·mL^-1.The limit of quantification was 0.5 ng·mL^-1~10 ng·mL^-1.The recovery was 62.4%~97.4%,and the matrix effect was 82.8%~109%.The standard deviation was less than 10.3%.The correlation coefficient was greater than 0.99,and the linear relationship was in the range of 10 ng·mL^-1~1000 ng·mL^-1.The method has high sensitivity and can screen,qualitatively and quantitatively analyze 29 pesticide components in blood,which can meet the requirements of pesticide component detection in actual blood samples.