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An efficient analytical method for determination of S-phenylmercapturic acid in urine by HPLC fluorimetric detector to assessing benzene exposure

Mendes, Michele P. Rocha ; Silveira, Josianne Nicácio ; Andre, Leiliane Coelho

Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, 2017-09, Vol.1063, p.136-140 [Peer Reviewed Journal]

Netherlands: Elsevier B.V

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  • Title:
    An efficient analytical method for determination of S-phenylmercapturic acid in urine by HPLC fluorimetric detector to assessing benzene exposure
  • Author: Mendes, Michele P. Rocha ; Silveira, Josianne Nicácio ; Andre, Leiliane Coelho
  • Subjects: Acetylcysteine - analogs & derivatives ; Acetylcysteine - chemistry ; Acetylcysteine - urine ; Adult ; Analytical validation ; Benzene ; Benzene - analysis ; Benzene - metabolism ; Chromatography, High Pressure Liquid - methods ; Female ; Fluorometry - methods ; Humans ; Limit of Detection ; Linear Models ; Male ; Middle Aged ; Occupational Exposure - analysis ; Reproducibility of Results ; S-phenylmercapturic acid ; Young Adult
  • Is Part Of: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, 2017-09, Vol.1063, p.136-140
  • Description: Benzene is an important occupational and environmental contaminant, naturally present in petroleum and as by-product in the steel industry. Toxicological studies showed pronounced myelotoxic action, causing leukemic and others blood cells disorders. Assessing of benzene exposure is performed by biomarkers as trans, trans-muconic acid (AttM) and S-phenylmercapturic acid (S-PMA) in urine. Due to specificity of S-PMA, this biomarker has been proposed to asses lower levels of benzene in air. The aim of this study was to validate an analytical method for the quantification of S-PMA by High-Performance Liquid Chromatography with fluorometric detector. The development of an analytical method of S-PMA in urine was carried out by solid phase extraction (SPE) using C-18 phase. The eluated were submitted to water bath at 75°C and nitrogen to analyte concentration, followed by alkaline hydrolysis and derivatization with monobromobimane. The chromatography conditions were reverse phase C-18 column (240mm, 4mm and 5μm) at 35°C; acetonitrile and 0.5% acetic acid (50:50) as mobile phase with a flow of 0.8mL/min. The limits of detection and quantification were 0.22μg/L and 0.68μg/L, respectively. The linearity was verified by simple linear regression, and the method exhibited good linearity in the range of 10–100μg/L. There was no matrix effect for S-PMA using concentrations of 40, 60, 80 and 100μg/L. The intra- and interassay precision showed coefficient of variation of less than 10% and the recovery ranged from 83.4 to 102.8% with an average of 94.4%. The stability of S-PMA in urine stored at −20°C was of seven weeks. The conclusion is that this method presents satisfactory results per their figures of merit. This proposed method for determining urinary S-PMA showed adequate sensitivity for assessment of occupational and environmental exposure to benzene using S-PMA as biomarker of exposure.
  • Publisher: Netherlands: Elsevier B.V
  • Language: English
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