Occupational exposure to volatile organic compounds (VOC) might generate serious worker health damages. Therefore, biological monitoring is essential to evaluate exposure biomarkers from highly toxic chemicals, ensuring better attention to the worker health. In this study was developed and validated a bioanalytical method based on high-performance liquid chromatography coupled to photodiode array (HPLC-PDA) for the quantification of VOC biomarkers in urine samples from Federal University of Goias (UFG) workers. Samples were collected from 30 occupationally exposed subjects after application of a questionnaire survey. The following biomarkers hippuric acid, methyl-hippuric acid, mandelic acid, phenylglyoxylic acid and phenol were quantified, representing exposition to toluene, xylene, styrene, ethylbenzene, benzene and phenol solvents, respectively. Hippuric acid levels were found close to or above the reference values, although a subject had levels higher than preconized by Biological Limit Values (BLV) guideline of 4.0 mg/g creatinine. Five subjects had 3 and 4-methylhippuric acid ranging from 0.1 to 1.0 mg/g creatinine. These results indicate a moderate to high VOC exposure from UFG workers. Multivariate analysis generated four clusters and indicated that histotechnicians and graphic workers need especial attention on occupational VOC exposure. The results from this study reinforce the need for reliable methods able to the biological monitoring as an important tool for assessing occupational exposure.