Method for the determination of benzene metabolite t,t-Muconic acid in urine by HPLC-UV with an Ion exclusion column.
No Thumbnail Available
Date
2016
Journal Title
Journal ISSN
Volume Title
Publisher
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.
Description
Keywords
Exposure biomarker, Environmental exposure, Benzene
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: <http://www.mdpi.com/2297-8739/3/2/14>. Acesso em: 15 set. 2017.