Determination of hippuric acid and o-, m- and p-methylhippuric acids in urine by capillary gas chromatography.

A capillary gas chromatographic (GC) method for the simultaneous determination of urinary hippuric acid (HA) and o-, m- and p-methylhippuric acids (MHAs), metabolites of toluene and o-, m- and p-xylenes, respectively, is described. These metabolites are converted into their isopropyl derivatives by extractive alkylation with tetrahexylammonium ion as extracting agent and isopropyl bromide as alkylating reagent in benzene. The derivatives are analysed using a chromatograph equipped with hydrogen flame ionization detector, split injection system and DB-17 capillary column. Benzoylleucine is used as an internal standard. The derivatives are well separated within 5 min and no interfering peaks are observed. The calibration curves of HA and MHAs in the range 1-50 micrograms are linear and sufficiently reproducible for quantitative analysis. Urine can be analysed accurately and precisely by this method without prior clean-up of the sample.