Serum Triton-X 100 treatment does not affect individual serum bile acid levels

Abstract Objectives Triton X-100 is a nonionic surfactant used for solvent/detergent viral inactivation, disrupting lipid-enveloped viruses while preserving proteins. At 1 % in plasma, it inactivates SARS-CoV-2 within 10 min with ≥6 log 10 titer reduction and is compatible with many immunoassays. Because LC-MS/MS bile acid analysis relies on chromatographic resolution, detergents could alter analytes via mixed micelle formation and matrix effects, making the effect of Triton X-100 on bile acid measurement clinically important yet untested. Methods We assessed 1 % Triton X-100 on LC-MS/MS bile acid quantification in serum from three critically ill, SARS-CoV-2–negative adults. Serum aliquots were untreated or incubated with 1 % Triton X-100, spiked with deuterated internal standards, extracted with methanol, and analyzed by HPLC-ES-MS/MS using water and an acetonitrile/methanol mobile phase with 0.1 % formic acid and 20 mM ammonium acetate. Analytes were quantified by retention times, MRM transitions, and peak areas normalized to internal standards in SCIEX OS Software. Results and conclusions Bile acid concentrations in treated vs. untreated samples were compared using Mann-Whitney U and Bland-Altman analyses. Differences were minimal and not statistically significant, indicating that 1 % Triton X-100 inactivation preserves LC-MS/MS bile acid quantification while improving biosafety.