金玉娥, 徐骞, 薛黎明, 乐孙阳, 熊丽蓓, 唐艳, 万静宜, 汪国权. 亲水作用色谱结合超高效液相色谱-三重四级杆质谱法检测蔬菜水果中8种高极性农药残留[J]. 环境与职业医学, 2022, 39(6): 701-707. DOI: 10.11836/JEOM21439
引用本文: 金玉娥, 徐骞, 薛黎明, 乐孙阳, 熊丽蓓, 唐艳, 万静宜, 汪国权. 亲水作用色谱结合超高效液相色谱-三重四级杆质谱法检测蔬菜水果中8种高极性农药残留[J]. 环境与职业医学, 2022, 39(6): 701-707. DOI: 10.11836/JEOM21439
JIN Yu’e, XU Qian, XUE Liming, LE Sunyang, XIONG Libei, TANG Yan, WAN Jingyi, WANG Guoquan. Determination of eight high polar pesticide residues in vegetables and fruits by hydrophilic interaction liquid chromatography combined with ultra-performance liquid chromatography-tandem mass spectrometry[J]. Journal of Environmental and Occupational Medicine, 2022, 39(6): 701-707. DOI: 10.11836/JEOM21439
Citation: JIN Yu’e, XU Qian, XUE Liming, LE Sunyang, XIONG Libei, TANG Yan, WAN Jingyi, WANG Guoquan. Determination of eight high polar pesticide residues in vegetables and fruits by hydrophilic interaction liquid chromatography combined with ultra-performance liquid chromatography-tandem mass spectrometry[J]. Journal of Environmental and Occupational Medicine, 2022, 39(6): 701-707. DOI: 10.11836/JEOM21439

亲水作用色谱结合超高效液相色谱-三重四级杆质谱法检测蔬菜水果中8种高极性农药残留

Determination of eight high polar pesticide residues in vegetables and fruits by hydrophilic interaction liquid chromatography combined with ultra-performance liquid chromatography-tandem mass spectrometry

  • 摘要: 背景 高极性农药在我国的产量和用量均占世界第一,需要一个速度快、通量大、准确度高的方法来测定食品中该类农药残留。

    目的 利用超高效液相色谱-三重四级杆质谱,建立蔬菜水果中8种高极性农药(矮壮素、百草枯、野燕枯、灭蝇胺、霜霉威、草甘膦、氨甲基膦酸和草铵膦)残留的检测方法。

    方法 通过比较多种类型亲水作用色谱柱及对流动相pH值和缓冲液浓度的优化,实现8种农药有效色谱保留和分离。基于色谱条件下高极性农药的质谱优化,建立目标物的多反应监测(MRM)通道。在样品前处理中对基质含水量、提取溶剂、净化方式进行优化,采用超高效液相色谱-三重四级杆质谱的MRM模式检测,同位素内标法定量。以加标回收率评价方法的准确度,相对标准偏差评价方法的精密度。将所建立的方法应用于57件市售蔬菜水果样本的检测,考察方法的适应性和了解高极性农药的残留状况。

    结果 对电喷雾源正离子模式(ESI+)检测,选择Sielc Obelisc R为色谱柱,20 mmol·L−1甲酸铵水溶液(pH=3±0.05)和乙腈做流动相;对电喷雾源负离子模式(ESI)检测,选择Shodex Asahipak NH2P-50 2D为色谱柱,5 mmol·L−1乙酸铵水溶液(pH=11±0.05)和乙腈做流动相,可以获得较好的色谱分离和峰形;在对样品含水量标化,以2%酸化甲醇为提取溶剂,C18分散固相萃取净化的最优化条件下,矮壮素、百草枯、野燕枯、灭蝇胺、霜霉威5种农药在1.00~100 μg·L−1浓度范围内,草甘膦、氨甲基膦酸、草铵膦3种农药在5.00~500 μg·L−1浓度范围内线性良好(相关系数>0.999),方法的检出限为0.002~0.010 mg·kg−1,定量限为0.005~0.025 mg·kg−1。在低、中、高3个浓度(定量限、2倍定量限和5倍定量限)加标水平下,方法的回收率在85.3%~113.2%之间,相对标准偏差范围为1.5%~9.5%(n=6)。57件市售蔬菜水果样品中检出的农药有矮壮素、灭蝇胺和霜霉威,有1份豇豆样品的灭蝇胺残留量超过了最大残留限量。

    结论 本研究建立的亲水作用色谱结合超高效液相色谱-三重四级杆质谱,同位素内标定量的方法具有简单、稳定、易操作的特点,适用于大批量蔬菜水果中8种高极性农药残留的快速筛查和定量检测,可为高极性农药的残留监测和风险评估提供技术保障。

     

    Abstract: Background The production and consumption of high polar pesticides in China are the largest in the world. Therefore, it is urgent to develop a method with fast analysis, large flux, and high accuracy to determine the residues of these pesticides in food.

    Objective To establish a method for the determination of eight highly polar pesticides chlormequat, paraquat, difenzoquat, cyromazine, propamocarb, glyphosate, (aminomethyl)-phosphonic acid, and glufosinate in vegetables and fruits by ultra-performance liquid chromatography-tandem mass spectrometry.

    Methods After comparing various types of hydrophilic interaction liquid chromatography (HILIC) columns, and optimizing pH value and buffer concentration of mobile phase, effective chromatographic retention and separation of selected eight pesticides were achieved. Based on the optimization of mass spectrometry under chromatographic conditions, a multiple reaction monitoring (MRM) channel of target compounds was established. In the sample pretreatment, through optimization of water content, extraction solvent, and purification method, a final MRM mode of ultra-performance liquid chromatography-tandem mass spectrometry was used for detection, and the isotope internal standard method was used for quantification. The accuracy and the precision of the method were evaluated using recovery and relative standard deviation. The established method was applied to detect 57 samples of retail vegetables and fruits to investigate the adaptability of the proposed method and the residual levels of selected high polar pesticides.

    Results For positive ion electrospray ionization (ESI+) detection, we chose Sielc Obelisc R as chromatographic column, and 20 mmol·L−1 ammonium formate solution (pH=3±0.05) and acetonitrile as mobile phase; for negative ion electrospray ionization (ESI) detection, we chose Shodex Asahipak NH2P-50 2D as chromatographic column, and 5 mmol·L−1 ammonium acetate solution (pH=11±0.05) and acetonitrile as mobile phase to obtain good chromatographic separation and peak shape. Under the optimal conditions of sample water content standardization, using 2% acidified methanol as extraction solvent, and C18 dispersed solid phase extraction purification, the linearity ranges of five analytes (chlormequat, paraquat, difenzoquat, cyromazine, and propamocarb) and three analytes glyphosate, (aminomethyl)phosphonic acid, and glufosinate were 1.00-100 μg·L−1 and 5.00-500 μg·L−1 (both correlation coefficients>0.999) respectively, the detection limits were 0.002-0.010 mg·kg−1, and the limits of quantification (LOQ) were 0.005-0.025 mg·kg−1. At three spiked levels (LOQ, 2LOQ, and 5LOQ), the recoveries were in the range of 85.3%–113.2%, and the relative standard deviations were 1.5%–9.5% (n=6). Three target pesticides (chlormequat, cyromazine, and propamocarb) were detected in 57 samples of retail vegetables and fruits, and the residue of chlormequat in cowpea exceeded the maximum residue limit.

    Conclusion The established method of HILIC combined with ultra-performance liquid chromatography-tandem mass spectrometry and isotopic internal standard quantification has the characteristics of simplicity, stability, and easy operation, which is suitable for rapid screening and quantitative detection of selected eight high polar pesticide residues in large quantities of vegetables and fruits, and provides technical support for monitoring and risk assessment of high polar pesticide residues.

     

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