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Analysis of glyphosate, aminomethylphosphonic acid, and glufosinate from human urine by HRAM LC-MS

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Abstract

Aminomethylphosphonic acid (AMPA) is the main metabolite of glyphosate (GLYP) and phosphonic acids in detergents. GLYP is a synthetic herbicide frequently used worldwide alone or together with its analog glufosinate (GLUF). The general public can be exposed to these potentially harmful chemicals; thus, sensitive methods to monitor them in humans are urgently required to evaluate health risks. We attempted to simultaneously detect GLYP, AMPA, and GLUF in human urine by high-resolution accurate-mass liquid chromatography mass spectrometry (HRAM LC-MS) before and after derivatization with 9-fluorenylmethoxycarbonyl chloride (Fmoc-Cl) or 1-methylimidazole-sulfonyl chloride (ImS-Cl) with several urine pre-treatment and solid phase extraction (SPE) steps. Fmoc-Cl derivatization achieved the best combination of method sensitivity (limit of detection; LOD) and accuracy for all compounds compared to underivatized urine or ImS-Cl-derivatized urine. Before derivatization, the best steps for GLYP involved 0.4 mM ethylenediaminetetraacetic acid (EDTA) pre-treatment followed by SPE pre-cleanup (LOD 37 pg/mL), for AMPA involved no EDTA pre-treatment and no SPE pre-cleanup (LOD 20 pg/mL) or 0.2–0.4 mM EDTA pre-treatment with no SPE pre-cleanup (LOD 19–21 pg/mL), and for GLUF involved 0.4 mM EDTA pre-treatment and no SPE pre-cleanup (LOD 7 pg/mL). However, for these methods, accuracy was sufficient only for AMPA (101–105%), while being modest for GLYP (61%) and GLUF (63%). Different EDTA and SPE treatments prior to Fmoc-Cl derivatization resulted in high sensitivity for all analytes but satisfactory accuracy only for AMPA. Thus, we conclude that our HRAM LC-MS method is suited for urinary AMPA analysis in cross-sectional studies.

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The data that support the reported findings of this study are available from AAF and XL.

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Funding

This study received funding from the National Cancer Institute (P30 CA71789).

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Authors and Affiliations

  1. University of Hawaii Cancer Center, Analytical Biochemistry Shared Resource, 701 Ilalo Street, Honolulu, HI, 96813, USA

    Adrian A. Franke, Xingnan Li & Jennifer F. Lai

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  1. Adrian A. Franke
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The manuscript was written through contributions of all authors and all authors approved the final version.

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Correspondence to Adrian A. Franke.

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Deidentified urine samples in this study were obtained from pre- and post-menopausal women, who provided written, informed consent, from a study that was approved by the University of Hawaii Office of Research Compliance and has since been discontinued. Quality control samples were pooled according to menopausal status.

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Franke, A.A., Li, X. & Lai, J.F. Analysis of glyphosate, aminomethylphosphonic acid, and glufosinate from human urine by HRAM LC-MS. Anal Bioanal Chem 412, 8313–8324 (2020). https://doi.org/10.1007/s00216-020-02966-1

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