Creatine and 3,4-Dimethylhippuric acid (3,4-DMHA), a glycine conjugate of 1,2,4-trimethyl-benzene (1,2,4-TMB) were determined in the urine of workers exposed to 1,2,4-TMB vapor. The best condition for the simultaneous determination of 3,4-DMHA and creatine by high performance liquid chromatography was obtained by reverse phase $C_{18}$ column (4.6${ imes}$150mm, 5${mu}m$) as stationary phase and 20% acetonitrile in 20mM phosphate buffer (pH 3.0) containing 4mM sodium octylsulfate(SOS)as mobile phase. The recovery of 3,4-DMHA spiked to blank urine in the range of 1~5${mu}g$/ml was about 96%. The concentration of urinary 3,4-DMHA of workers had a positive correlation with the environmental level of 1,2,4-TMB (r=0.866). The data suggest that urinary 3,4-DMHA concentration is a useful biological index for 1,2,4-TMB exposure.