Validation of HPLC Method for Determination of trans,trans-Muconic Acid as Bio¬marker of Benzene Exposure

  • I. Bajusová Department of Occupational Medicine and Clinical Toxicology, L. Pasteur University Hospital, Košice,
  • Ľ. Legáth Department of Occupational Medicine and Clinical Toxicology of Faculty of Medicine, P. J. Šafárik University, Košice,
  • T. Gondová Department of Analytical Chemistry,
  • Z. Vargová Department of Inorganic Chemistry, Faculty of Science, P. J. Šafárik University, Košice, Slovakia


Determination of phenol in urine is a widely used method. However, it may provide false positive results due to possible presence of aromatic amino acids, which can also produce phenol in urine. Trans,trans-muconic acid (MA) is a non-phenolic metabolite of benzene which appears in urine. The present review deals with determination of MA by RP-HPLC with UV detection. MA was isolated from urine by solid phase extraction (SPE). The mean recovery was 96103 %. The detection and quantification limits were 0.057 mg l1 and 0.189 mg l1, respectively. This method can be used to determine, control and monitor the level of MA in urine of workers, who are overexposed to benzene. The obtained data were compared with those of the non-exposed control group. The results suggest that MA could be one of the validated benzene bio¬markers in risk assessment.