High performance liquid chromatography for determination of urinary metabolites of toluene, xylene and styrene and its application.
TL;DRAbstract
The purpose of this research was to determine mandelic, phenylglyoxylic, hippuric, o-, m- and p-methylhippuric acids, the six urinary metabolites of styrene, toluene and xylene by high performance liquid chromatography (HPLC). These metabolites were extracted in an acid medium, transferred into a basic solution and back extracted again using ethyl acetate, and the organic phase was evaporated to dryness under a compressed air flow at room temperature. The residue obtained was dissolved in 1 ml mobile phase solution of 0.01 M potassium orthophosphate in 0.3% acetic acid (adjusted to a pH of 2.5 with orthophosphoric acid):tetrahydrofuran:acetonitrile(87:5:8) and 100 microl was injected into a HPLC equipped with a 4.6 x 250 mm ODS3-C18 reversed phased column and ultraviolet (UV) detector at a wavelength of 254 nm. All metabolites were clearly separated within 21 minutes. The detection limits of the method were 1.1 ng/ml for PGA, 4.9 ng/ml for HA, 17.0 ng/ml for MA, 2.5 ng/ml for o-MHA, 1.
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The purpose of this research was to determine mandelic, phenylglyoxylic, hippuric, o-, m- and p-methylhippuric acids, the six urinary metabolites of styrene, toluene and xylene by high performance liquid chromatography (HPLC). These metabolites were extracted in an acid medium, transferred into a basic solution and back extracted again using ethyl acetate, and the organic phase was evaporated to dryness under a compressed air flow at room temperature. The residue obtained was dissolved in 1 ml mobile phase solution of 0.01 M potassium orthophosphate in 0.3% acetic acid (adjusted to a pH of 2.5 with orthophosphoric acid):tetrahydrofuran:acetonitrile(87:5:8) and 100 microl was injected into a HPLC equipped with a 4.6 x 250 mm ODS3-C18 reversed phased column and ultraviolet (UV) detector at a wavelength of 254 nm. All metabolites were clearly separated within 21 minutes. The detection limits of the method were 1.1 ng/ml for PGA, 4.9 ng/ml for HA, 17.0 ng/ml for MA, 2.5 ng/ml for o-MHA, 1.
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