用于检测草甘膦的酶联免疫吸附测定法的开发。

Development of an enzyme-linked immunosorbent assay for the detection of glyphosate.

作者信息

Clegg B S, Stephenson G R, Hall J C

机构信息

Laboratory Services Division and Department of Environmental Biology, University of Guelph, Guelph, Ontario, Canada N1G 2W1.

出版信息

J Agric Food Chem. 1999 Dec;47(12):5031-7. doi: 10.1021/jf990064x.

DOI:10.1021/jf990064x

PMID:10606568

Abstract

A competitive indirect enzyme-linked immunosorbent assay (CI-ELISA) was developed to quantitate the herbicide glyphosate [N-(phosphonomethyl)glycine] in water. The ELISA has a detection limit of 7.6 microg mL(-1) and a linear working range of 10-1000 microg mL(-1) with an IC(50) value of 154 microg mL(-1). The glyphosate polyclonal antisera did not cross-react with a number of other herbicides tested but did cross-react with the glyphosate metabolite aminomethylphosphonic acid and a structurally related herbicide, glyphosine [(N,N-bis(phosphonomethyl)glycine]. The assay was used to estimate, quantitatively with accuracy and precision, glyphosate concentrations in water samples. Water samples were analyzed directly, and no sample preparation was required. To improve detection limits, water samples were concentrated prior to analysis, resulting in the increase of the detection limits by 100-fold. After the sample preconcentration step, the detection limit improved to 0.076 microg mL(-1) with an IC(50) value of 1.54 microg mL(-1), and a linear working range was 0.1-10 microg mL(-1). Glyphosate concentrations determined by ELISA correlated well with those determined by high-pressure liquid chromatography (r(2) = 0.99). This assay contributes to reducing the costs associated with conventional residue analysis techniques for the quantitation of glyphosate in water.

摘要

开发了一种竞争性间接酶联免疫吸附测定法（CI-ELISA）来定量水中的除草剂草甘膦[N-(膦酰基甲基)甘氨酸]。该ELISA的检测限为7.6 μg mL⁻¹，线性工作范围为10 - 1000 μg mL⁻¹，IC₅₀值为154 μg mL⁻¹。草甘膦多克隆抗血清与多种测试的其他除草剂无交叉反应，但与草甘膦代谢物氨甲基膦酸和一种结构相关的除草剂草甘膦[(N,N-双(膦酰基甲基)甘氨酸]有交叉反应。该测定法用于准确且精确地定量估计水样中的草甘膦浓度。水样可直接分析，无需样品制备。为提高检测限，水样在分析前进行浓缩，使检测限提高了100倍。经过样品预浓缩步骤后，检测限提高到0.076 μg mL⁻¹，IC₅₀值为1.54 μg mL⁻¹，线性工作范围为0.1 - 10 μg mL⁻¹。通过ELISA测定的草甘膦浓度与通过高压液相色谱法测定的浓度相关性良好（r² = 0.99）。该测定法有助于降低与用于定量水中草甘膦残留的传统分析技术相关的成本。