title: Comprehensive profiling of mercapturic acid metabolites from dietary acrylamide as short-term exposure biomarkers for evaluation of toxicokinetics in rats and daily internal exposure in humans using isotope dilution ultra-high performance liquid chromatography tandem mass spectrometry.
abstract: Mercapturic acid metabolites from dietary acrylamide are important short-term exposure biomarkers for evaluating the in vivo toxicity of acrylamide. Most of studies have focused on the measurement of two metabolites, N-acetyl-S-(2-carbamoylethyl)-L-cysteine (AAMA) and N-acetyl-S-(2-carbamoyl-2-hydroxyethyl)-L-cysteine (GAMA). Thus, the comprehensive profile of acrylamide urinary metabolites cannot be fully understood. We developed an isotope dilution ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method for the simultaneous determination of all four mercapturic acid adducts of acrylamide and its primary metabolite glycidamide under the electroscopy ionization negative (ESI-) mode in the present study. The limit of detection (LOD) and limit of quantification (LOQ) of the analytes ranged 0.1-0.3 ng/mL and 0.4-1.0 ng/mL, respectively. The recovery rates with low, intermediate and high spiking levels were calculated as 95.5%-105.4%, 98.2%-114.0% and 92.2%-108.9%, respectively. Acceptable within-laboratory reproducibility (RSD<7.0%) substantially supported the use of current method for robust analysis. Rapid pretreatment procedures and short run time (8 min per sample) ensured good efficiency of metabolism profiling, indicating a wide application for investigating short-term internal exposure of dietary acrylamide. Our proposed UHPLC-MS/MS method was successfully applied to the toxicokinetic study of acrylamide in rats. Meanwhile, results of human urine analysis indicated that the levels of N-acetyl-S-(2-carbamoylethyl)-L-cysteine-sulfoxide (AAMA-sul), which did not appear in the mercapturic acid metabolites in rodents, were more than the sum of GAMA and N-acetyl-S-(1-carbamoyl-2-hydroxyethyl)-L-cysteine (iso-GAMA). Thus, AAMA-sul may alternatively become a specific biomarker for investigating the acrylamide exposure in humans. Current proposed method provides a substantial methodology support for comprehensive profiling of toxicokinetics and daily internal exposure evaluations of acrylamide in vivo.
authors.affiliation: Department of Food Science and Nutrition, College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, Zhejiang, China; Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang R & D Center for Food Technology and Equipment, Fuli Institute of Food Science, Zhejiang University, Hangzhou 310058, Zhejiang, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Food Science and Nutrition, College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, Zhejiang, China.
authors.forename: Qiao
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Department of Food Science and Nutrition, College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, Zhejiang, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Cheng
authors.affiliation: Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051, Zhejiang, China.
authors.forename: Jingshun
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051, Zhejiang, China.
authors.forename: Jiaojiao
authors.initials: J
authors.lastname: Xu
authors.affiliation: Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051, Zhejiang, China. Electronic address: renyiping@263.net.
authors.forename: Yiping
authors.initials: Y
authors.lastname: Ren
mesh_terms: D000111:Acetylcysteine / Q000737:chemistry* / Q000633:toxicity / Q000652:urine; D020106:Acrylamide / Q000737:chemistry*; D000818:Animals; D015415:Biomarkers / Q000032:analysis* / Q000737:chemistry; D002623:Chemistry Techniques, Analytical; D002851:Chromatography, High Pressure Liquid; D004032:Diet; D005260:Female; D006801:Humans; D007554:Isotopes / Q000737:chemistry*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D053719:Tandem Mass Spectrometry; D066007:Toxicokinetics; D055815:Young Adult
chemical_list: D015415:Biomarkers; D007554:Isotopes; D020106:Acrylamide; D000111:Acetylcysteine
keywords: Acrylamide; Glycidamide; Internal exposure; Mercapturic acid metabolites; Toxicokinetics; Urinary biomarkers