Assay of short-chain carboxylic acids in plasma and urine using gas chromatography after extractive alkylation.

Abstract

After extractive alkylation combined with plasma deproteinization, we used gas chromatography to assay for short-chain carboxylic acids from formic acid to valeric acid in plasma and urine. It was possible to provide highly sensitive analysis with 0.1-3.4µg/mL as the limit of detection for plasma and 0.6-8.0µg/mL for urine, with a correlation coefficient of 1.000 for the linear regression calibration curves. For plasma, deproteinization using ultrafiltration before extractive alkylation resulted in a higher sensitivity for acetic, propionic, butyric, and valeric acids compared with the method without deproteinization. The concentrations of formic acid and acetic acid were determined to be 6µg/mL and 10µg/mL, respectively, in the tested plasma, and 22µg/mL and 32µg/mL, respectively, in the tested urine. Concentrations from propionic acid to valeric acid were ≤ 1.3µg/mL. In addition, high concentrations of sulfate, phosphate, hydrogen carbonate, ammonium, and/or sodium ions did not remarkably inhibit the derivatization of carboxylic acids, although hydrogen carbonate ions significantly inhibited that of formic acid.