4.5 Article

Optimization and validation of a liquid chromatography-tandem mass spectrometry method for the determination of glyphosate in human urine after pre-column derivatization with 9-fluorenylmethoxycarbonyl chloride

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ELSEVIER
DOI: 10.1016/j.jchromb.2021.122616

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Glyphosate; Urine; FMOC-Cl; Derivatization; LC-MS/MS

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In 2015, glyphosate was classified as probably carcinogenic to humans by the International Agency for Research on Cancer, leading to increased public concerns about its environmental and health risks. A simple LC-MS/MS method was developed for quantification of glyphosate in human urine and successfully applied to 52 urine samples from exposed subjects in northern Argentina.
In 2015, glyphosate was classified as 'Group 2A' probably carcinogenic to humans? by the International Agency for Research on Cancer (IARC). Therefore, public concerns about the environmental and health risks of this substance have rapidly increased. Considering its toxicokinetic characteristics, urinary levels of glyphosate could be a powerful tool for human biomonitoring. Nevertheless, the physicochemical properties of this molecule and the complexity of the matrix make this purpose particularly challenging. In order to solve this problem, the presented study describes a simple LC-MS/MS method for the quantification of glyphosate in human urine after pre-column derivatization with FMOC-Cl. Method development was focused on the optimization of the derivatization reaction in human urine, adjusting critical variables such as pH of borate buffer, FMOC-Cl concentration and derivatization time. Besides, chromatographic separation and spectrometric parameters were also established. The analytical method was fully validated according international guidelines for selectivity, carry over, linearity, accuracy, precision, lower limit of quantitation, matrix effect and stability under different conditions. All performance parameters were within the acceptance criteria. In addition, the method was successfully applied to 52 urine samples obtained from exposed subjects from northern Argentina, laying the foundation for future epidemiological studies.

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