High throughput HPLC-ESI(-)-MS/MS methodology for mercapturic acid metabolites of 1,3-butadiene: Biomarkers of exposure and bioactivation.
1,3-Butadiene (BD) is an important industrial and environmental carcinogen present in cigarette smoke, automobile exhaust, and urban air. The major urinary metabolites of BD in humans are 2-(N-acetyl-L-cystein-S-yl)-1-hydroxybut-3-ene/1-(N-acetyl-L-cystein-S-yl)-2-hydroxybut-3-ene (MHBMA), 4-(N-acetyl-L-cystein-S-yl)-1,2-dihydroxybutane (DHBMA), and 4-(N-acetyl-L-cystein-S-yl)-1,2,3-trihydroxybutyl mercapturic acid (THBMA), which are formed from the electrophilic metabolites of BD, 3,4-epoxy-1-butene (EB), hydroxymethyl vinyl ketone (HMVK), and 3,4-epoxy-1,2-diol (EBD), respectively. In the present work, a sensitive high-throughput HPLC-ESI(-)-MS/MS method was developed for simultaneous quantification of MHBMA and DHBMA in small volumes of human urine (200 μl). The method employs a 96 well Oasis HLB SPE enrichment step, followed by isotope dilution HPLC-ESI(-)-MS/MS analysis on a triple quadrupole mass spectrometer. The validated method was used to quantify MHBMA and DHBMA in urine of workers from a BD monomer and styrene-butadiene rubber production facility (40 controls and 32 occupationally exposed to BD). Urinary THBMA concentrations were also determined in the same samples. The concentrations of all three BD-mercapturic acids and the metabolic ratio (MHBMA/(MHBMA+DHBMA+THBMA)) were significantly higher in the occupationally exposed group as compared to controls and correlated with BD exposure, with each other, and with BD-hemoglobin biomarkers. This improved high throughput methodology for MHBMA and DHBMA will be useful for future epidemiological studies in smokers and occupationally exposed workers.
Department of Medicinal Chemistry, University of Minnesota, Minneapolis, MN 55455, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA.
Srikanth
S
Kotapati
Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA; Department of Chemistry, University of Minnesota, Minneapolis, MN 55455, USA.
Amanda
A
Esades
Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA.
Brock
B
Matter
Department of Biostatistics, University of Minnesota, Minneapolis, MN 55455, USA.
Chap
C
Le
Department of Medicinal Chemistry, University of Minnesota, Minneapolis, MN 55455, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA. Electronic address: trety001@umn.edu.
Natalia
N
Tretyakova
D000111:Acetylcysteine / Q000737:chemistry*; D015415:Biomarkers / Q000737:chemistry* / Q000652:urine*; D002070:Butadienes / Q000737:chemistry* / Q000633:toxicity / Q000652:urine*; D002074:Butanones / Q000737:chemistry / Q000652:urine; D002273:Carcinogens / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D020724:Elastomers / Q000633:toxicity; D004852:Epoxy Compounds / Q000737:chemistry / Q000652:urine; D006801:Humans; D016273:Occupational Exposure / Q000032:analysis*; D013343:Styrenes / Q000633:toxicity; D053719:Tandem Mass Spectrometry / Q000379:methods; D014556:Urine / Q000737:chemistry
D015415:Biomarkers; D002070:Butadienes; D002074:Butanones; D002273:Carcinogens; D020724:Elastomers; D004852:Epoxy Compounds; D013343:Styrenes; C495785:hydroxymethylvinyl ketone; C024370:3,4-epoxy-1-butene; C065815:styrene-butadiene rubber; C031763:1,3-butadiene; D000111:Acetylcysteine
1,3-Butadiene; Metabolism; Occupational exposure; Quantitative analysis; Urinary metabolites