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Title: Method for the determination of benzene metabolite t,t-Muconic acid in urine by HPLC-UV with an Ion exclusion column.
Authors: Gomes, Rafaela de Paiva
Sanson, Ananda Lima
Lobo, Fabiana Aparecida
Afonso, Robson José de Cássia Franco
Coutrim, Maurício Xavier
Keywords: Exposure biomarker
Environmental exposure
Issue Date: 2016
Citation: GOMES, R. de P. et al. Method for the determination of benzene metabolite t,t-Muconic acid in urine by HPLC-UV with an Ion exclusion column. Separations, v. 3, n. 2, p. 14, 2016. Disponível em: <>. Acesso em: 15 set. 2017.
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 g 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 g 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 g 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.
ISSN: 2297-8739
metadata.dc.rights.license: This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution License ( Fonte: o próprio artigo.
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