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Monitoring exposure to acrylamide by the determination of S-(2-carboxyethyl)cysteine in hydrolyzed hemoglobin by gas chromatography-mass spectrometry
Authors:E Bailey  P B Farmer  I Bird  J H Lamb  J A Peal
Institution:1. Department of Pediatric Surgery, Division of Pediatric Urology, Ege University, Izmir, Turkey;2. Department of Urology, Ege University, Izmir, Turkey;3. Department of Pathology, Ege University, Turkey;1. Saint Joseph University, Faculty of Sciences, Chemistry Department, B.P. 11-514, 11072050, Lebanon;2. “arcenciel”, Environment Program, B.P. 165216, Beirut, Lebanon;3. Ecole Nationale Supérieure des Ingénieurs en Arts Chimiques Et Technologiques, Toulouse, France;1. Chemical and Biological Engineering Department, South Dakota School of Mines & Technology, Rapid City, SD, 57701, USA;2. Department of Chemical and Life Science Engineering, Virginia Commonwealth University, 601 W. Main St., Richmond, VA, 23284, USA;3. Department of Civil and Environmental Engineering, Old Dominion University, 135 Kaufman Hall, Norfolk, VA, 23529, USA;4. Bioenergy Technologies Department, Clean Energy & Transportation Division, Idaho National Laboratory, Idaho Falls, ID, 83415, USA;1. Department of Ecology and Environmental Protection, Faculty of Environmental and Mechanical Engineering, Poznań University of Life Sciences, Piatkowska 94C, 60-649, Poznań, Poland;2. Department of Mathematical and Statistical Methods, Faculty of Agronomy, Horticulture and Bioengineering, Poznań University of Life Sciences, Wojska Polskiego 28, 60-637, Poznań, Poland;3. Department of Land Improvement, Environmental Development and Spatial Management, Faculty of Environmental and Mechanical Engineering, Poznań University of Life Sciences, Piatkowska 94E, 60-649, Poznań, Poland;4. Institute of Environmental Protection and Development, Faculty of Environmental Engineering and Geodesy, Wroc?aw University of Environmental and Life Sciences, Plac Grunwaldzki 24, 50-363, Wroc?aw, Poland;5. Department of Remote Sensing and Environmental Assessment, Institute of Environmental Engineering, Warsaw University of Life Sciences, Nowoursynowska 166, 02-787, Warsaw, Poland;6. Department of Chemistry, Faculty of Forestry and Wood Technology, Poznań University of Life Sciences, Wojska Polskiego 75, 60-625 Poznań, Poland;7. Department of Analytical Chemistry, Faculty of Chemistry, Adam Mickiewicz University, Uniwersytetu Poznańskiego 8, 61-614, Poznań, Poland;1. Laboratory of Pharmacology, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, China;2. State Key Laboratory of Coordination Chemistry, Nanjing University, Nanjing, China;3. Department of Scientific Research, Shanghai Public Health Clinical Center, Fudan University, Shanghai, China;4. Department of Cardiovascular Diseases, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, China;5. Shimadzu Biomedical Research Laboratory, Shanghai, China;6. Department of Pharmacology, Jiangsu Jiankang Vocational College, Nanjing, China
Abstract:Acrylamide is a potent cumulative neurotoxin in animals and man. In vivo exposure to this electrophile results in the formation of a covalently bound reaction product with cysteine residues in hemoglobin. This adduct yields on acid hydrolysis S-(2-carboxyethyl)cysteine which has been analyzed by capillary gas chromatography with mass spectrometry. Globin isolated from the blood of rats exposed to acrylamide was spiked with an internal standard (globin treated in vitro with d3-acrylamide) and was then hydrolyzed with 6 N HCl. The protein hydrolysate was fractionated on a Dowex 50W H+ ion exchange column and the amino acids in the partially purified extract were determined as N-heptafluorobutyryl methyl esters using an OV-1701 fused silica capillary column. Quantitation was made by chemical ionization (isobutane) selective ion monitoring in which the ions m/z 386 (M-OCH3)+ derived from derivatized S-(2-carboxyethyl)cysteine in the sample and the corresponding ion m/z 389 from the added deuterium-labeled internal standard were monitored. The dose-response relationship between production of hemoglobin adduct and dose of acrylamide (0.1 mg/kg-5 mg/kg) is curved, showing an increasing slope with increasing doses of acrylamide.
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