Archives of Toxicology

, Volume 62, Issue 4, pp 247–253 | Cite as

Hydroxyethylvaline adduct formation in haemoglobin as a biological monitor of cigarette smoke intake

  • Eric Bailey
  • Alan G. F. Brooks
  • Colin T. Dollery
  • Peter B. Farmer
  • Barry J. Passingham
  • Marcus A. Sleightholm
  • David W. Yates
Original Investigations

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.

Key words

Hydroxyethylvaline Haemoglobin Cigarette smoke Biological monitoring Cotinine carcinogen adduct 

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References

  1. 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: 145–152Google Scholar
  2. 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: 187–191Google Scholar
  3. Bryant MS, Skipper PL, Tannenbaum SR, Maclure M (1987) Hemoglobin adducts of 4-aminobiphenyl in smokers and nonsmokers. Cancer Res 47: 602–608Google Scholar
  4. Binder H, Lindner W (1972) Bestimmung von Äthylenoxid im Rauch garantiert unbegaster Zigaretten. Fachliche Mitt Austria Tabakwerke AG 13: 215–220Google Scholar
  5. 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: 427–442Google Scholar
  6. 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: 283–293Google Scholar
  7. 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: 79–87Google Scholar
  8. Davis RA (1986) The determination of nicotine and cotinine in plasma. J Chromatogr Sci 24: 134–141Google Scholar
  9. Doll R, Hill AB (1950) Smoking and carcinoma of the lung. Preliminary report. Br Med J ii: 739–748Google Scholar
  10. 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: 83–103Google Scholar
  11. 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: 175–184Google Scholar
  12. Elmenhorst H, Schultz C (1968) Flüchtige Inhaltsstoffe des Tabakrauches. Beitr Tabakforsch 4: 90–122Google Scholar
  13. 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: 637–640Google Scholar
  14. Farmer PB, Neuman HG, Henschler D (1987) Estimation of exposure of man to substances reacting covalently with macromolecules. Arch Toxicol 60: 251–260Google Scholar
  15. 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: 337–344Google Scholar
  16. 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 LyonsGoogle Scholar
  17. 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: 335–339Google Scholar
  18. 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: 37–41Google Scholar
  19. Neumann HG (1984) Analysis of haemoglobin as a dose monitor for alkylating and arylating agents. Arch Toxicol 56: 1–6Google Scholar
  20. 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: 1–10Google Scholar
  21. 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: 395–405Google Scholar
  22. 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: 449–456Google Scholar
  23. 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: 1377–1380Google Scholar
  24. Remmer H (1987) Passively inhaled tobacco smoke: a challenge to toxicology and preventive medicine. Arch Toxicol 61: 89–104Google Scholar
  25. 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: 731–738Google Scholar
  26. Segerbäck D (1983) Alkylation of DNA and hemoglobin in the mouse following exposure to ethene and ethene oxide. Chem Biol Interact 45: 139–151Google Scholar
  27. 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: 255–266Google Scholar
  28. 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: 1519–1521Google Scholar
  29. 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: 1217–1222Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • Eric Bailey
    • 1
  • Alan G. F. Brooks
    • 1
  • Colin T. Dollery
    • 2
  • Peter B. Farmer
    • 1
  • Barry J. Passingham
    • 1
  • Marcus A. Sleightholm
    • 2
  • David W. Yates
    • 3
  1. 1.MRC Toxicology UnitMRC LaboratoriesCarshaltonUK
  2. 2.Department of MedicineRoyal Postgraduate Medical School, Hammersmith HospitalLondonUK
  3. 3.Hope HospitalSalfordUK

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