Gianfranco Frigerio1, Rosa Mercadante2, Elisa Polledri3, Pasquale Missineo4, Laura Campo5, Silvia Fustinoni6. 1. EPIGET - Epidemiology, Epigenetics, and Toxicology Lab, Department of Clinical Sciences and Community Health, University of Milan and Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, via S. Barnaba, 8, 20122 Milan, Italy. Electronic address: gianfranco.frigerio@unimi.it. 2. EPIGET - Epidemiology, Epigenetics, and Toxicology Lab, Department of Clinical Sciences and Community Health, University of Milan and Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, via S. Barnaba, 8, 20122 Milan, Italy. Electronic address: rosa.mercadante@unimi.it. 3. EPIGET - Epidemiology, Epigenetics, and Toxicology Lab, Department of Clinical Sciences and Community Health, University of Milan and Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, via S. Barnaba, 8, 20122 Milan, Italy. Electronic address: elisa.polledri@policlinico.mi.it. 4. EPIGET - Epidemiology, Epigenetics, and Toxicology Lab, Department of Clinical Sciences and Community Health, University of Milan and Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, via S. Barnaba, 8, 20122 Milan, Italy. Electronic address: pasquale.missineo@policlinico.mi.it. 5. EPIGET - Epidemiology, Epigenetics, and Toxicology Lab, Department of Clinical Sciences and Community Health, University of Milan and Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, via S. Barnaba, 8, 20122 Milan, Italy. Electronic address: laura.campo@policlinico.mi.it. 6. EPIGET - Epidemiology, Epigenetics, and Toxicology Lab, Department of Clinical Sciences and Community Health, University of Milan and Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, via S. Barnaba, 8, 20122 Milan, Italy. Electronic address: silvia.fustinoni@unimi.it.
Abstract
INTRODUCTION: Mercapturic acids are urinary metabolites of occupational and environmental toxicants. The aim of this work was to develop and validate an analytical assay for the determination of several urinary mercapturic acids to be used as biomarkers of exposure. METHOD: An isotope dilution tandem mass spectrometric method, coupled with reversed-phase liquid chromatography, was developed for the analysis of mercapturic acids derived from several compounds, including those of benzene, toluene, 1,3-butadiene, styrene, acrylonitrile, 4-chloronitrobenzene, acrylamide, acrolein, propylene oxide, N,N-dimethylformamide, crotonaldehyde, ethylene oxide, and methylating and ethylating agents. Samples were prepared by simple filtration after dilution. A validation was carried out, including the assessment of calibration curves, sensitivity, accuracy, precision, process efficiency, and stability, along with external verification. The assay was applied to the analysis of 14 end-of-shift urine samples from unexposed workers and gasoline station attendants. RESULTS: The chromatographic run lasted 18 min. Limits of quantitation ranged from 0.01 to 3.2 μg/L; precision, expressed as relative standard deviation, ranged from 0.6 to 20.9%; and accuracy ranged from 93.4 to 114.9% of theoretical values. The use of deuterated internal standards was suitable for control of the matrix effect. The assay allowed the simultaneous quantitation of urinary mercapturic acids at different concentration ranges. The external verification exercise produced good results. The application of the assay to urine samples from workers revealed differences in mercapturic acid profiles in agreement with the expected patterns of exposure. CONCLUSION: This high-throughput method is valid and useful for the quantitation of urinary mercapturic acids, and is suitable for human biomonitoring of occupational and environmental exposure.
INTRODUCTION:Mercapturic acids are urinary metabolites of occupational and environmental toxicants. The aim of this work was to develop and validate an analytical assay for the determination of several urinary mercapturic acids to be used as biomarkers of exposure. METHOD: An isotope dilution tandem mass spectrometric method, coupled with reversed-phase liquid chromatography, was developed for the analysis of mercapturic acids derived from several compounds, including those of benzene, toluene, 1,3-butadiene, styrene, acrylonitrile, 4-chloronitrobenzene, acrylamide, acrolein, propylene oxide, N,N-dimethylformamide, crotonaldehyde, ethylene oxide, and methylating and ethylating agents. Samples were prepared by simple filtration after dilution. A validation was carried out, including the assessment of calibration curves, sensitivity, accuracy, precision, process efficiency, and stability, along with external verification. The assay was applied to the analysis of 14 end-of-shift urine samples from unexposed workers and gasoline station attendants. RESULTS: The chromatographic run lasted 18 min. Limits of quantitation ranged from 0.01 to 3.2 μg/L; precision, expressed as relative standard deviation, ranged from 0.6 to 20.9%; and accuracy ranged from 93.4 to 114.9% of theoretical values. The use of deuterated internal standards was suitable for control of the matrix effect. The assay allowed the simultaneous quantitation of urinary mercapturic acids at different concentration ranges. The external verification exercise produced good results. The application of the assay to urine samples from workers revealed differences in mercapturic acid profiles in agreement with the expected patterns of exposure. CONCLUSION: This high-throughput method is valid and useful for the quantitation of urinary mercapturic acids, and is suitable for human biomonitoring of occupational and environmental exposure.
Authors: Denise S Tevis; Sharon R Flores; Brandon M Kenwood; Deepak Bhandari; Peyton Jacob; Jia Liu; Pawel K Lorkiewicz; Daniel J Conklin; Stephen S Hecht; Maciej L Goniewicz; Benjamin C Blount; Víctor R De Jesús Journal: Int J Hyg Environ Health Date: 2021-05-04 Impact factor: 7.401
Authors: Louise Gren; Annette M Krais; Eva Assarsson; Karin Broberg; Malin Engfeldt; Christian Lindh; Bo Strandberg; Joakim Pagels; Maria Hedmer Journal: Int Arch Occup Environ Health Date: 2022-03-16 Impact factor: 2.851
Authors: Annette M Krais; Julie Y Essig; Louise Gren; Carolina Vogs; Eva Assarsson; Katrin Dierschke; Jörn Nielsen; Bo Strandberg; Joakim Pagels; Karin Broberg; Christian H Lindh; Anders Gudmundsson; Aneta Wierzbicka Journal: Int J Environ Res Public Health Date: 2021-06-16 Impact factor: 3.390