Simultaneous determination of urinary hippuric acid, o-, m- and p-methylhippuric acids, mandelic acid and phenylglyoxylic acid for biomonitoring of volatile organic compounds by gas chromatography–mass spectrometry

Abstract

A sensitive and precise method for the simultaneous determination of hippuric acid, o-, m- and p-methylhippuric acids, mandelic acid and phenylglyoxylic acid, which are major urinary metabolites of toluene, o-, m- and p-xylenes, styrene and ethylbenzene, respectively, was developed. These metabolites were converted into their methyl ester derivatives with methanol in hydrochloric acid, and then quantitated by gas chromatography–mass spectrometry (GC–MS) with selected ion monitoring using a DB-1 capillary column. The injected compounds were quantitatively and reproducibly resolved within 19 min with a detection limit of 8–27 pg. The calibration curves were linear in the range of 0.05–25 μg for each compound, with correlation coefficients above 0.9999. This method was successfully used to analyze small amounts of both rat and human urine samples without any interference from coexisting substances. Overall recoveries of these compounds spiked in urine samples were 92–104%. The analytical results of the contents of these metabolites in the rat and human urine samples are presented.