Abstract
The ethylene oxide adduct formed on the N-terminal valine in haemoglobin was investigated as a biological monitor of tobacco smoke intake. The modified method developed for the determination of the hydroxyethylvaline adduct (HOEtVal) involved reaction of globin with pentafluorophenyl isothiocyanate, extraction of the HOEtVal thiohydantoin product, derivatization of this by trimethylsilylation and quantitation by capillary gas chromatography with selective ion monitoring mass spectrometry using a tetradeuterated internal standard. The method was applied to globin samples from 26 habitual cigarette smokers and 24 non-smokers. There was a significant correlation between cigarette smoke intake as measured by the average number of cigarettes smoked per day and HOEtVal levels (r=0.537, p<0.01). Background levels were found in non-smokers (mean 49.9 pmol/g Hb, range 22–106 pmol/g Hb). Smoking increased these levels by 71 pmol/g Hb/ 10 cigarettes per day.
Cotinine levels in plasma of the smokers were determined by GC-NPD using 2-methyl-4-nitroaniline as internal standard. For non-smokers cotinine was determined by GC-MS selective ion monitoring using d3-methylcotinine as internal standard. There was no correlation between number of cigarettes smoked per day and cotinine levels (r=0.297, p>0.05) although cotinine was correlated with HOEtVal (r=0.43, p<0.01).
The HOEtVal adduct levels thus appear to be a suitable biomonitor for exposure to hydroxyethylating agents in cigarette smoke, reflecting an integrated dose over the erythrocyte lifetime. This is in contrast to plasma cotinine determinations which reflect only the previous day's exposure to nicotine in smoke.
Similar content being viewed by others
References
Bailey E, Farmer PB, Lamb JH (1980) The enantiomer as internal standard for the quantitation of the alkylated amino acid S-methyl-l-cysteine in hemoglobin by gas chromatographychemical ionisation mass spectrometry with single ion detection. J Chromatogr 200: 145–152
Bailey E, Farmer PB, Shuker DEG (1987) Estimation of exposure to alkylating carcinogens by the GC-MS determination of adducts to hemoglobin and nucleic acid bases in urine. Arch Toxicol 60: 187–191
Bryant MS, Skipper PL, Tannenbaum SR, Maclure M (1987) Hemoglobin adducts of 4-aminobiphenyl in smokers and nonsmokers. Cancer Res 47: 602–608
Binder H, Lindner W (1972) Bestimmung von Äthylenoxid im Rauch garantiert unbegaster Zigaretten. Fachliche Mitt Austria Tabakwerke AG 13: 215–220
Calleman CJ, Ehrenberg L, Jansson B, Osterman-Golkar S, Segerbäck D, Svensson K, Wachtmeister CA (1978) Monitoring and risk assessment by means of alkyl groups in hemoglobin in persons occupationally exposed to ethylene oxide. J Environ Pathol Toxicol 2: 427–442
Curvall M, Kazemi-Vala E, Enzell CR (1982) Simultaneous determination of nicotine and cotinine in plasma using capillary column gas chromatography with nitrogen-sensitive detection. J Chromatogr 232: 283–293
Daenens P, Lamelle L, Callewaertz K, De Schepper P, Galeazzi R, Van Rossum J (1985) Determination of cotinine in biological fluids by capillary gas chromatography — mass spectrometry — selected-ion monitoring. J Chromatogr 342: 79–87
Davis RA (1986) The determination of nicotine and cotinine in plasma. J Chromatogr Sci 24: 134–141
Doll R, Hill AB (1950) Smoking and carcinoma of the lung. Preliminary report. Br Med J ii: 739–748
Ehrenberg L, Hiesche KD, Osterman-Golkar S, Wennberg I (1974) Evaluation of genetic risks of alkylating agents: tissue doses in the mouse from air contaminated with ethylene oxide. Mutat Res 24: 83–103
Ehrenberg L, Osterman-Golkar S, Segerbäck D, Svensson K, Calleman CJ (1977) Evaluation of genetic risks of alkylating agents. III. Alkylation of haemoglobin after metabolic conversion of ethene to ethene oxide in vivo. Mutat Res 45: 175–184
Elmenhorst H, Schultz C (1968) Flüchtige Inhaltsstoffe des Tabakrauches. Beitr Tabakforsch 4: 90–122
Farmer PB, Bailey E, Grof SM, Törnqvist M, Osterman-Golkar S, Kautiainen A, Lewis-Enright DP (1986) Monitoring human exposure to ethylene oxide by the determination of haemoglobin adducts using gas chromatography-mass spectrometry. Carcinogenesis 7: 637–640
Farmer PB, Neuman HG, Henschler D (1987) Estimation of exposure of man to substances reacting covalently with macromolecules. Arch Toxicol 60: 251–260
Frank H, Hintze T, Bimboes D, Remmer H (1980) Monitoring lipid peroxidation by breath analysis: endogenous hydrocarbons and their metabolic elimination. Toxicol Appl Pharmacol 56: 337–344
International Agency for Research on Cancer (1986) Tobacco smoking. IARC monographs on the evaluation of the carcinogenic risks of chemicals to humans, vol. 38: IARC Lyons
Jarvis MJ, Tunstall-Pedoe H, Feyerabend C, Vesey C, Saloojee Y (1984) Biochemical markers of smoke absorption and self reported exposure to passive smoking. J Epidemiol Commun Health 38: 335–339
Muranaka H, Higashi E, Itani S, Shimizu Y (1988) Evaluation of nicotine, cotinine, thiocyanate, carboxyhemoglobin and expired carbon monoxide as biochemical tobacco smoke uptake parameters. Int Arch Occup Environ Health 60: 37–41
Neumann HG (1984) Analysis of haemoglobin as a dose monitor for alkylating and arylating agents. Arch Toxicol 56: 1–6
Osterman-Golkar S, Ehrenberg L, Segerbäck D, Hällström I (1976) Evaluation of genetic risk of alkylating agents. II. Hemoglobin as a dose monitor. Mutat Res 34: 1–10
Osterman-Golkar S, Farmer PB, Segerbäck D, Bailey E, Calleman CJ, Svensson K, Ehrenberg L (1983) Dosimetry of ethylene oxide in the rat by quantitation of alkylated histidine in hemoglobin. Teratogen Carcinogen Mutagen 3: 395–405
Perera FP, Santella RM, Brenner D, Poirier MC, Munshi AA, Fischman HK, Van Ryzin J (1987) DNA adducts, protein adducts and sister chromatid exchange in cigarette smokers and nonsmokers. JNCI 79: 449–456
Pojer R, Whitfield KB, Poulos V, Eckhard IF, Richmond R, Hensley W (1984) Carboxyhaemoglobin, cotinine and thiocyanate assay compared for distinguishing smokers from nonsmokers. Clin Chem 30: 1377–1380
Remmer H (1987) Passively inhaled tobacco smoke: a challenge to toxicology and preventive medicine. Arch Toxicol 61: 89–104
Sagai M, Ichinose T (1980) Age-related changes in lipid peroxidation as measured by ethane, ethylene, butane and pentane in respired gases of rats. Life Sci 27: 731–738
Segerbäck D (1983) Alkylation of DNA and hemoglobin in the mouse following exposure to ethene and ethene oxide. Chem Biol Interact 45: 139–151
Törnqvist M, Mowrer J, Jensen S, Ehrenberg L (1986a) Monitoring of environmental cancer initiators through hemoglobin adducts by a modified Edman degradation method. Anal Biochem 154: 255–266
Törnqvist M, Osterman-Golkar S, Kautiainen A, Jensen S, Farmer PB, Ehrenberg L (1986b) Tissue doses of ethylene oxide in cigarette smokers determined from adduct levels in hemoglobin. Carcinogenesis 7: 1519–1521
Wald NJ, Nanchahal K, Thompson SG, Cuckle HS (1986) Does breathing other people's tobacco smoke cause lung cancer? Contemporary themes. Br Med J 293: 1217–1222
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bailey, E., Brooks, A.G.F., Dollery, C.T. et al. Hydroxyethylvaline adduct formation in haemoglobin as a biological monitor of cigarette smoke intake. Arch Toxicol 62, 247–253 (1988). https://doi.org/10.1007/BF00332482
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00332482