Direct determination of t,t-muconic acid in human urine by two-dimensional liquid chromatography

Abstract

The accurate determination of E,E-2,4-hexadiendioic acid (t,t-muconic acid), an important urinary biomarker of benzene exposure, is directly performed in human urine by a new two-dimensional liquid chromatographic method. After a first partial separation of urine components by a reversed-phase mechanism and the focalisation of the anionic analytes on a small anion-chromatographic column, a conventional ion-chromatographic analysis with UV detection is carried out. The analytical procedure is fully controlled by the HPLC instrumentation software using an eight-port switch valve. If compared with conventional one-dimensional procedures, the new method produces chromatograms containing a limited number of well resolved peaks and consequently allows better analytical performances: no interfering peaks, absence of bias, repeatability better than 5% in the concentration range 0.09–5 mg dm⁻³ and a detection limit of 4.0 μg dm⁻³ (α = β = 0.05) for the analysis of real samples.