色谱

色谱 ›› 2025, Vol. 43 ›› Issue (2): 155-163.DOI: 10.3724/SP.J.1123.2024.03008

• 研究论文 • 上一篇    下一篇

通过式固相萃取-超高效液相色谱-串联质谱法测定沉积物中草铵膦、草甘膦及其代谢物

杨霄1,2, 谢仲桂1,2, 李小玲1,2, 索纹纹1,2, 陈相艺1,2, 万译文1,2,*()   

  1. 1.湖南省水产科学研究所, 湖南 长沙 410153
    2.农业农村部渔业产品质量检验测试中心(长沙), 湖南 长沙 410153
  • 收稿日期:2024-03-08 出版日期:2025-02-08 发布日期:2025-01-23
  • 通讯作者: *E-mail:cnjs3385@163.com.
  • 基金资助:
    湖南省科技创新计划项目(2023NK4168);国家现代农业产业技术体系建设项目(CARS-48-39)

Determination of glufosinate, glyphosate and their metabolites in sediment by ultra-high performance liquid chromatography-tandem mass spectrometry with pass-through solid-phase extraction

YANG Xiao1,2, XIE Zhonggui1,2, LI Xiaoling1,2, SUO Wenwen1,2, CHEN Xiangyi1,2, WAN Yiwen1,2,*()   

  1. 1. Hunan Fisheries Science Institute, Changsha 410153, China
    2. Fishery Products Quality Testing Center, Ministry of Agriculture and Rural Affairs, Changsha 410153, China
  • Received:2024-03-08 Online:2025-02-08 Published:2025-01-23
  • Supported by:
    Science and Technology Innovation Program of Hunan Province(2023NK4168);Earmarked Fund for China Agriculture Research System(CARS-48-39)

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摘要：

建立了通过式固相萃取-超高效液相色谱-串联质谱法测定沉积物中草铵膦、草甘膦及其6种代谢物(3-甲基磷酸亚基丙酸、N-乙酰草铵膦、氨甲基膦酸、N-乙酰氨甲基膦酸、N-乙酰草甘膦、N-甲基草甘膦)的方法。样品采用4%(体积分数)氨水溶液提取,提取液经PRiME HLB固相萃取柱净化,过0.22 μm聚醚砜滤膜后供超高效液相色谱-串联质谱测定。目标化合物使用Metrosep A Supp 5阴离子色谱柱(150 mm×4.0 mm, 5 μm)分离,以水和200 mmol/L碳酸氢铵溶液(含0.05%(v/v)氨水)作为流动相进行梯度洗脱,在电喷雾离子源(ESI)、负离子扫描和多反应监测(MRM)模式下进行测定,基质匹配外标法定量。结果表明,草铵膦、草甘膦及其代谢物在15 min内即可完成色谱分离,色谱峰形良好,响应值高,目标化合物在2.0~200.0 μg/L范围内线性关系良好,相关系数均大于0.995。草铵膦、3-甲基磷酸亚基丙酸、N-乙酰草铵膦、N-乙酰氨甲基膦酸、N-乙酰草甘膦、N-甲基草甘膦的检出限为5 μg/kg,定量限为20 μg/kg;草甘膦、氨甲基膦酸的检出限为10 μg/kg,定量限为30 μg/kg。以空白沉积物为基质样品,在3个加标水平(定量限、5倍定量限和10倍定量限)下,低有机质含量的沉积物中目标化合物的平均回收率为78.5%~107%,相对标准偏差为1.32%~14.7%(n=6);高有机质含量的沉积物中目标化合物的平均回收率为76.4%~113%,相对标准偏差为2.60%~11.2%(n=6)。采用本方法对池塘、湖泊、水库、河流等不同类型的沉积物样品进行测定,结果显示,湖泊、水库、河流沉积物样品中未检出草铵膦、草甘膦及其代谢物,1个池塘沉积物样品中检出草甘膦和氨甲基膦酸,检出含量分别为31.7 μg/kg和52.3 μg/kg。本研究建立的方法具有简单、快速、绿色环保、准确度和灵敏度高、重复性好等优势,适用于沉积物中草铵膦、草甘膦及其代谢物的快速检测,为研究其在沉积物中的残留特征和环境行为提供了技术支持。

关键词: 超高效液相色谱-串联质谱, 通过式固相萃取, 草铵膦, 草甘膦, 代谢物, 沉积物, 非衍生化

Abstract:

Glufosinate (GLUF) and glyphosate (GLY) are nonselective phosphorus-containing amino acid herbicides that are widely used in agricultural gardens and noncultivated areas. These herbicides give rise to a number of key metabolites, with 3-methyl phosphinicopropionic acid (MPPA), N-acetyl glufosinate (N-acetyl GLUF), aminomethyl phosphonic acid (AMPA), N-acetyl aminomethyl phosphonic acid (N-acetyl AMPA), N-acetyl glyphosate (N-acetyl GLY), N-methyl glyphosate (N-methyl GLY) as the major metabolites obtained from GLUF and GLY. Extensive use of these herbicides may lead to their increased presence in the environment, especially aquatic ecosystems. An increasing number of research studies into the toxicities of GLUF, GLY, and their metabolites have shown that these herbicides are potentially toxic to aquatic biota. GLUF and GLY, as well as their metabolites, are extremely polar and water-soluble, and they lack chromogenic and fluorescent groups; therefore, their concentrations are difficult to determine using conventional methods. Most analytical methods used to date have largely depended on derivatization procedures, leading to overall determination processes that are tedious and time-consuming. Therefore, establishing a quick and sensitive method that does not require derivatives for determining GLUF, GLY, and their metabolites in water environments, including surface water, sediments, and aquatic organisms, is an important endeavor.

In this study, a new approach was developed based on pass-through solid-phase extraction coupled with ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) to determine GLUF, GLY, and their metabolites, including MPPA, N-acetyl GLUF, AMPA, N-acetyl AMPA, N-acetyl GLY, and N-methyl GLY, in sediments. Samples were extracted with 4% (v/v) ammonia water and purified using PRiME HLB pass-through solid-phase extraction columns. The extracts were filtered through a polyethersulfone microfiltration membrane and analyzed by UHPLC-MS/MS. Compounds were separated on a Metrosep A Supp 5 column (150 mm×4.0 mm, 5 μm) using gradient elution with water and 200 mmol/L ammonium hydrogen carbonate solution containing 0.05% (v/v) ammonia water as the mobile phases. Analytes were detected using MS/MS with a negative electrospray ionization (ESI-) source in the multiple reaction monitoring (MRM) mode. A matrix-matched external-standard approach was used for quantitative analysis. GLUF, GLY, and their metabolites were detected within 15 min with good peak shapes and high responses. Calibration curves were linear in the range of 2.0-200 μg/L, with correlation coefficients exceeding 0.995. This method delivered limits of detection (LODs) and limits of quantification (LOQs) of 5 μg/kg and 20 μg/kg, respectively, for GLUF, MPPA, N-acetyl-GLUF, N-acetyl AMPA, N-acetyl GLY, and N-methyl GLY, and 10 μg/kg and 30 μg/kg, for GLY and AMPA, respectively. Average spiked recoveries at three levels (LOQ, 5LOQ, 10LOQ) in sediment with low organic matter content were in the range of 78.5%-107%, and the relative standard deviations (RSDs) were in the range of 1.32%-14.7% (n=6). Average spiked recoveries of 76.4%-113% were determined for three levels (LOQ, 5LOQ, and 10LOQ) in sediments with high organic matter contents, with RSDs of 2.60%-11.2% (n=6). The developed method was used to analyze GLUF, GLY, and their metabolites in ponds, lakes, reservoirs, and river sediments. No target compounds were detected in any sediment sample obtained from a lake, reservoir, or river; however, GLY and AMPA were detected in one pond-sediment sample at levels of 31.7 and 52.3 μg/kg, respectively. The developed approach is simple, fast, and green; moreover, it offers advantages, including high accuracies, high sensitivities, and good reproducibilities. Accordingly, the developed method is suitable for determining GLUF, GLY, and their metabolites in sediments and can provide technical support for studying residue characteristics and environmental behavior in sediments.

Key words: ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), pass-through solid-phase extraction, glufosinate, glyphosate, metabolites, sediment, non-derivatization

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