Rapid determination of polyethoxylated tallow amine surfactants in human plasma by LC-MSMS

The potential exposure to the widely used glyphosate-based herbicides, including attempted suicide by ingestion, is of world-wide concern. Whilst the major focus to date has been on managing exposure to the active ingredient glyphosate, it is now recognised that a common major 'inert' surfactant ingredient, polyethoxylated tallow amine (POEA) and related compounds, may be more toxic. However, the information on the toxicokinetics of POEA surfactants after exposure is limited, in part, due to the lack of suitable methods for their analysis in biological matrices. We therefore developed and validated a robust LC-MSMS method that allowed, for the first time, a rapid analysis of 11 POEA homologues in human plasma. Chromatographic separation was achieved on a Kinetex EVO C18 column under a 5 min gradient elution with mobile phase A containing water/acetonitrile/formic acid (95:5:0.1, v/v/v) and mobile phase B containing acetonitrile/water/formic acid (95:5:0.1, v/v/v). Amlodipine was chosen as the internal standard (IS) for this assay. Amlodipine-d4 would be an ideal alternative IS to expand the applicability of the established method especially in antihypertensive patients. Multiple reaction monitoring (MRM) methods were optimized for 11 POEA homologues and the IS. Sample pre-treatment was performed using simple protein precipitation with methanol at a ratio of 4:1, requiring only 20 μL plasma. The validated method showed good specificity, accuracy and precision with lower limits of quantification (LLOQ) ranging from 0.35 to 10.8 ng mL^-1 for all selected POEA homologues. The method was then used to measure concentrations of the various POEA surfactants in more than 600 human plasma samples from 151 patients admitted to hospital with acute glyphosate intoxication. The highest concentrations ranged from 1.07 ng mL^-1 for C₁₈u(EO)₄-362.70 ng mL^-1 for C₁₆s(EO)₂. The analysis of POEA surfactants plasma concentrations as described here underpins the assessment of POEA internal exposure and the relationships between POEA related glyphosate toxicity and the extent of poisoning.