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Separations 2016, 3(2), 14; doi:10.3390/separations3020014

Method for the Determination of Benzene Metabolite t,t-Muconic Acid in Urine by HPLC-UV with an Ion Exclusion Column

Department of Chemistry, Institute of Exact and Biological Sciences, Federal University of Ouro Preto, Campus Morro do Cruzeiro, s/n, Bauxita, 35400-000 Ouro Preto, MG, Brazil
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Author to whom correspondence should be addressed.
Academic Editor: Frank L. Dorman
Received: 14 December 2015 / Revised: 6 April 2016 / Accepted: 26 April 2016 / Published: 13 May 2016
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Abstract

The benzene metabolite trans,trans-muconic acid (tt-MA) is widely used as a biological indicator of exposure to this xenobiotic. An analytical method was developed and validated for the determination of urinary tt-MA using solid phase extraction onto a Strata SAX cartridge. The analysis was performed by HPLC with an Aminex HPX-87H ion exclusion column and a UV diode array detector. The sample preparation conditions were optimized using a 24−1 factorial design. Comparison of the slopes of standard analytical curves prepared in aqueous solution and urine showed that the biological matrix suppressed the tt-MA signal by around 50%, so the analytical curves were prepared with tt-MA standard at low concentrations in pooled urine. The analytical curves in the range of 5–500 μg·L−1 showed determination coefficients values (R2) > 0.99 for tt-MA standard in water and R2 > 0.98 for tt-MA standards in pooled urine. The coefficients of variation obtained using seven replicates were lower than 3.6%, and recoveries of tt-MA from solutions containing 5, 25, 50 and 100 μg·L−1 of the analyte were in the range 85%–90%, demonstrating the satisfactory precision and accuracy of the method. The limits of detection and quantification were 0.11 and 0.36 µg ·L−1, respectively. The benefits of this new method developed is the possibility of complete chromatographic peak separation for the determination of tt-MA at baseline, without matrix components’ interference as normally found in the C18 column. This is the first time that this chromatographic column has been used for the analysis of tt-MA in urine. View Full-Text
Keywords: benzene; trans,trans-muconic acid; exposure biomarker; environmental exposure; Aminex HPX-87H ion exclusion column benzene; trans,trans-muconic acid; exposure biomarker; environmental exposure; Aminex HPX-87H ion exclusion column
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Gomes, R.P.; Sanson, A.L.; Lobo, F.A.; Afonso, R.J.C.F.; Coutrim, M.X. Method for the Determination of Benzene Metabolite t,t-Muconic Acid in Urine by HPLC-UV with an Ion Exclusion Column. Separations 2016, 3, 14.

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