HPLC-MS/MS结合液液萃取法检测尿样中22种新烟碱类农药及其代谢物

Determination of 22 Neonicotinoid Pesticides and Their Metabolites in Urine Samples by HPLC-MS/MS Combined with Liquid-Liquid Extraction

建立了高效液相色谱-串联质谱(HPLC-MS/MS)同时检测尿液中22种新烟碱类农药(NEOs)及其代谢物的分析方法。尿样经酶解,甲酸调节pH值,液液萃取法提取,HSS T3色谱柱分离后,以(0.1%甲酸+5mmol/L甲酸铵)/水-乙腈作为流动相进行梯度洗脱,正/负离子模式切换扫描,多反应监测(MRM)模式检测,内标法定量。结果表明,22种NEOs及其代谢物在各自质量浓度范围内呈良好线性关系(r^(2)>0.995 0),方法检出限和定量下限分别为0.05~0.10 ng/mL和0.15~0.30 ng/mL;在低、中、高3个加标水平下,平均回收率为64.5%~125%,相对标准偏差(RSD)为0.10%~12%。样品稳定性实验表明,尿样采集后应尽快检测。利用该方法对54份真实尿样进行测定,均有不同程度的检出。所建立的方法高效、稳定,可实现22种NEOs及代谢物的同时检测。

An analytical method based on high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)was established for the simultaneous detection of 22 neonicotinoid pes⁃ticides(NEOs)and their metabolites in human urine.Urine was extracted by liquid-liquid extraction after enzymatic hydrolysis,pH adjusted by formic acid,and then separated by HSS T3 column.Gra⁃dient elution was performed with(0.1%formic acid+5 mmol/L ammonium formate)/water-acetoni⁃trile as mobile phase.The samples were scanned simultaneously in positive/negative ion mode,de⁃tected in multiple reaction monitoring(MRM)mode,and quantized by internal standard method.The results showed that there was a good linear relationship between the 22 NEOs and their metabo⁃lites in the mass concentration ranges(r^(2)>0.9950).The detection limit and quantitation limit of the method were 0.05-0.10 ng/mL and 0.15-0.30 ng/mL,respectively.The average recoveries were 64.5%-125%at low,middle and high spiked levels.The relative standard deviations ranged from 0.10%to 12%.In addition,sample stability tests indicated that urine samples should be tested as soon as possible after collection.The method had been used to determine 54 real urine samples.All samples were detected NEOs or their metabolites.The established method is efficient and stable,and can simultaneously detect 22 NEOs and their metabolites.