Sensitive high-performance liquid chromatography method for the simultaneous determination of low levels of dichloroacetic acid and its metabolites in blood and urine

Abstract

Dichloroacetic acid (DCA) is a contaminant found in treated drinking water due to chlorination. DCA has been shown to be a complete hepatocarcinogen in both mice and rats. In this study we developed a rapid and sensitive high-performance liquid chromatography (HPLC) method to simultaneously detect DCA and its metabolites, oxalic acid, glyoxylic acid and glycolic acid in blood and urine samples of animals sub-chronically administered with DCA (2 g/l) in drinking water. Both urine and plasma samples were treated minimally before HPLC analysis. Separation and detection of DCA and its metabolites were achieved using an anion-exchange column and a conductivity detector. The mobile phase consisted of an initial concentration of 0.01 m M sodium hydroxide in 40% methanol followed by a linear gradient from 0.01 m M to 60 m M sodium hydroxide in 40% methanol for 30 min. The lower detection limit for DCA and each of its three major metabolites was 0.05 μg/ml. DCA and its metabolites gave a linear response range from 0.05 to 100 μg/ml. Plasma DCA was also analyzed by gas chromatography (GC), and the results obtained correlated with those from the HPLC method (correlation coefficient=0.999). While available HPLC techniques offer sensitive procedures to detect either glycolic acid or oxalic acid, the described HPLC method has the unique advantage of determining simultaneously the parent compound (DCA) and its three major metabolites (oxalic acid, glyoxylic acid and glycolic acid) in biological samples, without complex sample preparation.