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Clinical Chemistry 39: 1788-1792, 1993;
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Clinical Chemistry, Vol 39, 1788-1792, Copyright © 1993 by American Association for Clinical Chemistry

Urinary trans,trans-muconic acid determined by liquid chromatography: application in biological monitoring of benzene exposure

BL Lee, AL New, PW Kok, HY Ong, CY Shi and CN Ong
WHO Collaborating Centre on Occupational Health, Department of Community Medicine, National University of Singapore, Kent Ridge.

We describe a sensitive and specific high-performance liquid- chromatographic method for determining the benzene metabolite, trans,trans-muconic acid (ttMA) in urine by measuring ultraviolet absorbance at 265 nm. We mix 1 mL of urine sample with 2 mL of Tris buffer containing vanillic acid as internal standard (IS) and percolate this through a preconditioned ion-exchange column. After rinsing the column with phosphoric acid solution, acetate buffer, and deionized water, we elute the analytes with 2 mL of an equivolume solution of 1.5 mol/L sodium chloride and methanol. Of this, 5 microL is injected into the HPLC column. The mobile phase used consists of, per liter, 10 mL of acetic acid, 100 mL of methanol, and the rest 5 mmol/L sodium acetate. The flow rate was started at 1 mL/min and increased to 1.5 mL/min after 6 min. ttMA and IS were detected at 5.2 and 10.2 min, respectively. The lowest detection limit is 125 pg. Analytical recovery and reproducibility generally exceeded 90%. We validated the method with urine samples collected from normal persons and from refinery workers exposed to benzene concentrations < 1 microL/L. The results show that urinary ttMA is a promising biological marker for risk assessment of low-concentration benzene exposure.


The following articles in journals at HighWire Press have cited this article:


Home page
 Annals of Clinical & Laboratory ScienceHome page
V. Wiwanitkit, J. Suwansaksri, and P. Nasuan
Urine Trans,trans-muconic Acid as a Biomarker for Benzene Exposure in Gas Station Attendants in Bangkok, Thailand
Ann. Clin. Lab. Sci., October 1, 2001; 31(4): 399 - 401.
[Abstract] [Full Text] [PDF]


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