Abstract
Styrene oxide, which is the genotoxically active metabolite of styrene, reacts in vivo with carboxylic acid residues in hemoglobin forming phenylhydroxyethyl esters. Mild alkali hydrolysis cleaves these ester adducts, yielding styrene glycol, which in human blood labelled in vitro with14C-styrene oxide accounted for 15% of the total radioactivity covalently bound to the protein. A quantitative assay procedure has been developed for measuring the base released styrene glycol in globin. The method utilizes solvent extraction followed by trimethylsilyl ether derivatization and separation and quantitation by capillary gas chromatography with selective ion recording mass spectrometry. Globin labelled in vitro with d8-styrene oxide was used as the internal standard. The method was used to establish a dose-response relationship in rats given single i.p. doses of styrene oxide (83.3–833 μmol/kg body wt). The method, which allows quantitation of the adducts down to levels of 15 pmol/g globin, has the potential to act as a dosimeter for industrial workers exposed to styrene or styrene oxide.
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Andersson HC, Tranberg EA, Uggla AH, Zetterberg G (1980) Chromosomal aberrations and sister chromatid exchanges in lymphocytes of men occupationally exposed to styrene in a plastic boat factory. Mutat Res 73: 387–401
Anson ML, Mirsky ME (1930) Protein coagulation and its reversal. The preparation of insoluble globin, soluble globin and heme. J Gen Physiol 13: 469–478
Bailey E, Brooks AG, Bird I, Fanner PB, Street B (1989) Monitoring exposure to 4,4′-methylene dianiline by the gas chromatographymass spectrometry determination of adducts to hemoglobin. Anal Biochem 190: 175–181
Bechtold WE, Sun JD, Wolff RK, Griffith WC, Kilmer JW, Bond JA (1991) Globin adducts of benzo(a)pyrene: markers of inhalation exposure as measured in F344/N rats. J Appl Toxicol 11: 115–118
Brenner DD, Jeffrey AM, Latriano L, Wazneh L, Warburton D, Toor M, Pero RW, Andrews LR, Walles S, Perera FP (1991) Biomarkers in styrene-exposed boat builders. Mutat Res 261: 225–236
Byfält Nordqvist M, Lof A, Osterman-Golkar S, Walles SAS (1985) Covalent binding of styrene and styrene 7,8-oxide to plasma proteins, hemoglobin and DNA in the mouse. Chem Biol Interact 55: 63–73
Camurri L, Codeluppi S, Pedroni C, Scarduelli L (1983) Chromosomal aberrations and sister chromatid exhanges in workers exposed to styrene. Mutat Res 119: 361–369
Day BW, Naylor S, Gan L-S, Sahali Y, Nguyen TT, Skipper PL, Wishnok JS, Tannenbaum SR (1990) Molecular dosimetry of polycyclic aromatic hydrocarbon epoxides and diol epoxides via hemoglobin adducts. Cancer Res 50: 4611–4618
Day BW, Naylor S, Gan L-S, Sahali Y, Ngugen TT, Skipper PL, Wishnok JS, Tannenbaum SR (1991) Gas chromatographic-mass spectrometric analysis of diols and tetrols from reactions of polycyclic aromatic hydrocarbon epoxides with hemoglobin. J Chromatogr 562: 563–571
ECETOC Monograph No. 13 (1989) DNA and protein adducts: evaluation of their use in exposure monitoring and risk assessment
Ehrenberg L, Osterman-Golkar S (1980) Alkylation of macromolecules for detecting mutagenic agents. Teratogen Carcinogen Mutagen 1: 105–127
Farmer PB, Bailey E (1989) Protein-carcinogen adducts in human dosimetry. Arch Toxicol Suppl. 13: 83–90
Farmer PB, Neumann HG, Henschler D (1987) Estimation of exposure of man to substances reacting covalently with macromolecules. Arch Toxicol 60: 251–260
Gan L-S, Wishnok JS, Fox JG, Tannenbaum SR (1989) Quantitation of methylated hemoglobin via hydrolysis of methyl esters to yield methanol. Anal Biochem 179: 326–331
Guillemin M, Bauer D (1976) Human exposure to styrene: II. Quantitative and specific gas Chromatographic analysis of urinary mandelic and phenylglyoxylic acids as an index of styrene exposure. Int Arch Occup Environ Health 37: 57–64
Hemminki K (1983) Reactions of methylnitrosourea, epichlorohydrin, styrene oxide and acetoxyacetylaminofluorene with polyamino acids. Carcinogenesis 4: 1–3
Hemminki K (1986) Binding of styrene oxide to amino acids, human serum proteins and hemoglobin. Arch Toxicol Suppl 9: 286–290
Hogstedt B, Hedner K, Mark-Vendel E, Mitelman F, Schutz A, Skerfving S (1979) Increased frequency of chromosome aberrations in workers exposed to styrene. Scand J Work Environ Health 5: 333–335
Kaur S, Hollander D, Haas R, Burlingame AL (1989) Characterisation of structural xenobiotic modifications in proteins by high sensitivity tandem mass spectrometry. Human hemoglobin treated in vitro with styrene 7,8-oxide. J Biol Chem 264: 16981–16984
Kostova M, Rizov N, Bencher I (1989) Gas Chromatographic method for determination of phenylglyoxylic and mandelic acid in urine of occupationally exposed workers to styrene. Khig Zdraveopaz 32: 86–90
Lijinsky W (1986) Rat and mouse forestomach tumors induced by chronic oral administration of styrene oxide. J Natl Cancer Inst 77: 471–476
Liu SF, Rappaport SM, Dongracz K, Bodell WJ (1988) Detection of styrene oxide-DNA adducts in lymphocytes of a worker exposed to styrene. In: Methods for detecting DNA damaging agents in humans. Applications in cancer epidemiology and prevention. IARC Scientific Publication No. 89, IARC, Lyon, pp 217–222
de Meester C, Poncelet F, Roberfroid M, Rondelet J, Mercier H (1977) Mutagenicity of styrene and styrene oxide. Mutat Res 56: 147–152
Neumann H-G (1988) Biomonitoring of aromatic amines and alkylating agents by measuring hemoglobin adducts. Int Arch Occup Environ Health 60: 151–155
Norppa H, Sorsa M, Pfaffli P, Vainio H (1980) Styrene and styrene oxide induce SCEs and are metabolized in human lymphocyte cultures. Carcinogenesis 1: 357–361
Norppa H, Hemminki K, Sorsa M, Vainio H (1981) Effect of monosubstituted epoxides on chromosome aberrations and SCE in cultured human lymphocytes. Mutat Res 91: 243–250
Pongracz K, Kaur S, Burlingame AL, Bodell WJ (1989) O6-substituted-2′-deoxyguanosine-3′-phosphate adducts detected by32P-postlabelling of styrene oxide treated DNA. Carcinogenesis 10: 1009–1013
Pongracz K, Kaur S, Burlingame AL, Bodell WJ (1992) Identification of N2-substituted 2′-deoxyguanosine-3′-phosphate adducts detected by32P-postlabelling of styrene-oxide-treated DNA. Carcinogenesis 13: 315–319
Ponomarkov V, Cabral JRP, Wahrendorf J, Galendo D (1984) A carcinogenicity study of styrene-7,8 oxide in rats. Cancer Lett 24: 95–101
Savela K, Hesso A, Hemminki K (1986) Characterization of reaction products between styrene oxide and deoxynucleosides and DNA. Chem Biol Interact 60: 235–246
Sollenberg J, Bjurström R, Wrangskog K, Vesterberg O (1988) Biological exposure limits estimated from relations between occupational styrene exposure during a work week and excretion of mandelic acid and phenylglyoxylic acids in urine. Int Arch Occup Environ Health 60: 365–370
Tates AD, Grummt T, Törnqvist M, Farmer PB, van Dam FJ, van Mossel H, Schoemaker HM, Osterman-Golkar S, Webel Ch, Tang YS, Zwinderman AH, Natarajan AT, Ehrenberg L (1991) Biological and chemical monitoring of occupational exposure to ethylene oxide. Mutat Res 250: 483–497
Ting D, Smith MT, Doane-Setzer P, Rappaport SM (1990) Analysis of styrene oxide-globin adducts based upon reaction with Raney nickel. Carcinogenesis 11: 755–760
Wade DR, Airy SC, Sinsheimer IE (1978) Mutagenicity of aliphatic epoxides. Mutat Res 58: 217–223
WHO (1983) Styrene: Environmental Health Criteria 26: World Health Organization, Geneva
Wigaeus E, Löf A, Bjurström R, Byfalt Nordqvist M (1983) Exposure to styrene. Uptake, distribution, metabolism and elimination in man. Scand J Work Environ Health 9: 476–488
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Sepai, O., Anderson, D., Street, B. et al. Monitoring of exposure to styrene oxide by GC-MS analysis of phenylhydroxyethyl esters in hemoglobin. Arch Toxicol 67, 28–33 (1993). https://doi.org/10.1007/BF02072031
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DOI: https://doi.org/10.1007/BF02072031