Archives of Toxicology

, Volume 64, Issue 3, pp 169–176 | Cite as

Reduction of hexavalent chromium by ascorbic acid and glutathione with special reference to the rat lung

  • Yasutomo Suzuki
  • Kazuo Fukuda
Original Articles


The reduction of 20 μM hexavalent chromium [chromium(VI)] byl-ascorbic acid (AsA) (0.06–2 mM) and/or glutathione (GSH) (2–15 mM) in buffer solutions, cell-free bronchoalveolar lavage fluids or soluble fractions of rat lungs was investigated at physiological pH (37° C). The reduction in AsA solution was pseudo-first-order in a single phase with respect to chromium(VI), but that in GSH solution showed a two-phase process. The half-life of chromium(IV) ranged from seconds to hours. The reducing ability of AsA was markedly higher than that of GSH. Coexistence of equimolar GSH with AsA accelerated the reduction rate slightly, in comparison with that in the corresponding AsA solution. Lavage fluids containing 0.06 mM AsA showed pH-dependent reactions similar to those of the corresponding AsA solutions. The lungsoluble fractions reduced chromium(VI) in a process composed of phase I and phase II, characterized by the reducing ability of AsA-GSH cooperation and of AsA alone, respectively. Reduction in the former was 30–40% more rapid than in the latter. The biological half-life of chromium(VI) in the lung was estimated to be 0.6 min, on the basis of the reducing activity in the first phase. However, the apparent biological half-life of chromium(VI) was about 2 min in rat lungs after intratracheal injection of chromate, involving depletion of AsA, but no significant changes in GSH. The difference is discussed in terms of AsA-induced initiative reduction in the alveolar lining fluid and subsequent obstructive effects of the resulting trivalent species on trans-membrane permeability of chromate anions. These results suggest that AsA is more reactive than GSH in the reduction of chromium(VI) in the rat lung and that the extracellular AsA in the alveolar lining fluid plays an important role in antioxidant defense against inhaled chromium(VI) compounds.

Key words

Hexavalent chromium Reduction Ascorbic acid Glutathione Rat lung Bronchoalveolar lavage fluid 


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  1. Adachi S, Yoshimura H, Miyayama R, Katayama H, Takemoto K, Kawai H (1983) Effects of chromium compounds on the respiratory system. Part 2. Difference between water-soluble hexavalent compounds and trivalent compounds. Jpn J Ind Health 25: 149–154Google Scholar
  2. Baldea I, Munteanu L (1980) The kinetics of the oxidation of ascorbic acid by chromate. Studia Univ Babes-Bolyai Chemia 25: 24–31Google Scholar
  3. Campomar JA, Fiol JJ, Terron A, Moreno V (1986) Chromium(III) interactions with nucleotides. II. Inorg Chim Acta 124: 75–81Google Scholar
  4. Cantin AM, North SL, Hubbard RC, Crystal RG (1987) Normal alveolar epithelial lining fluid contains high levels of glutathione. J Appl Physiol 63: 152–157PubMedGoogle Scholar
  5. Chatterjee IB (1973) Evolution and the biosynthesis of ascorbic acid. Science 182: 1271–1272PubMedGoogle Scholar
  6. Clark JH (1959) The denaturation of proteins by chromium salts. A M A Arch Ind Health 20: 117–123Google Scholar
  7. Connett PH, Wetterhahn KE (1983) Metabolism of the carcinogen chromate by cellular constituents. In: Structure and bonding. Inorganic elements in biochemistry. Springer-Verlag, Berlin, 54: 93–124Google Scholar
  8. Connett PH, Wetterhahn KE (1985) In vitro reaction of the carcinogen chromate with cellular thiols and carboxylic acids. J Am Chem Soc 107: 4282–4288Google Scholar
  9. DePamphilis ML, Cleland WW (1973) Preparation and properties of chromium(III)-nucleotide complexes for use in the study of enzyme mechanisms. Biochemistry 12: 3714–3724PubMedGoogle Scholar
  10. Franchini I, Magnani F, Mutti A (1983) Mortality experience among chromeplating workers. Scand J Work Environ Health 9: 247–252PubMedGoogle Scholar
  11. Garcia JD, Jennette KW (1981) Electron-transport cytochrome P-450 system is involved in the microsomal metabolism of the carcinogen chromate. J Inorg Biochem 14: 281–295PubMedGoogle Scholar
  12. Ginter E (1980) Endogenous ascorbic acid synthesis and recommended dietary allowances for vitamin C. Am J Clin Nutr 33: 1448–1449Google Scholar
  13. Gray SJ, Sterling K (1950) The tagging of red cells and plasma proteins with radioactive chromium. J Clin Invest 29: 1604–1613PubMedGoogle Scholar
  14. Gruber JE, Jennette KW (1978) Metabolism of the carcinogen chromate by rat liver microsomes. Biochem Biophys Res Commun 82: 700–706PubMedGoogle Scholar
  15. Hornig D (1975) Distribution of ascorbic acid, metabolites and analogues in man and animals. In: King CG, Burns JJ (ed) Second conference on vitamin C. Ann NY Acad Sci 258: 103–118Google Scholar
  16. Keeling PL, Smith LL (1982) Relevance of NADPH depletion and mixed disulphide formation in rat lung to the mechanism of cell damage following paraquat administration. Biochem Pharmacol 31: 3243–3249PubMedGoogle Scholar
  17. Keller DA, Menzel DB (1985) Picomole analysis of glutathione, glutathione disulfide, glutathione S-sulfonate, and cysteine S-sulfonate by high-performance liquid chromatography. Anal Biochem 151: 418–423PubMedGoogle Scholar
  18. Kitagawa S, Seki H, Kametani F, Sakurai H (1982) Uptake of hexavalent chromium by bovine erythrocytes and its interaction with cytoplasmic components; the role of glutathione. Chem Biol Interact 40: 265–274PubMedGoogle Scholar
  19. Korallus U, Harzdorf C, Lewalter J (1984) Experimental bases for ascorbic acid therapy of poisoning by hexavalent chromium compounds. Int Arch Occup Environ Health 53: 247–256PubMedGoogle Scholar
  20. Legg JI (1978) Substitution-inert metal ions as probes of biological function. Coordination Chem Rev 25: 103–132Google Scholar
  21. Léonard A, Lauwerys RR (1980) Carcinogenicity and mutagenicity of chromium. Mutat Res 76: 227–239PubMedGoogle Scholar
  22. Levis AG, Bianchi V (1982) Mutagenic and cytogenetic effects of chromium compounds. In: Langård S (ed) Biological and environmental aspects of chromium. Elsevier Biomedical Press, Amsterdam, New York, Oxford, pp 171–208Google Scholar
  23. Levis AG, Bianchi V, Tamino G, Pegoraro B (1978) Cytotoxic effects of hexavalent and trivalent chromium on mammalian cells in vitro. Br J Cancer 37: 386–396PubMedGoogle Scholar
  24. Mikalsen A, Alexander J, Ryberg D (1989) Microsomal metabolism of hexavalent chromium. Inhibitory effect of oxygen and involvement of cytochrome P-450. Chem Biol Interact 69: 175–192PubMedGoogle Scholar
  25. Nielson DW, Goerke J, Clements JA (1981) Alveolar subphase pH in the lungs of anesthetized rabbits. Proc Natl Acad Sci USA 78: 7119–7123PubMedGoogle Scholar
  26. Norseth T (1981) The carcinogenicity of chromium. Environ Health Perspect 40: 121–130PubMedGoogle Scholar
  27. Rucker RB, Dubick MA, Mouritsen J (1980) Hypothetical calculations of ascorbic acid synthesis based on estimates in vitro. Am J Clin Nutr 33: 961–964PubMedGoogle Scholar
  28. Samitz MH (1970) Ascorbic acid in the prevention and treatment of toxic effects from chromates. Acta Derm Venereol (Stockholm) 50: 59–64Google Scholar
  29. Scarpelli EM (1977) The surfactant system of the lung. Int Anesthesiol Clin 15: 19–60PubMedGoogle Scholar
  30. Stubbs DW, Griffin JF (1973) The influence of dietary protein on gulonolactone hydrolase, gulonate NADP oxidoreductase, and tissue ascorbate in male and female rats. Proc Soc Exp Biol Med 144: 199–202PubMedGoogle Scholar
  31. Suzuki Y (1988) Reduction of hexavalent chromium by ascorbic acid in rat lung lavage fluid. Arch Toxicol 62: 116–122PubMedGoogle Scholar
  32. Suzuki Y, Fukuda K (1989) Anion-exchange high-performance liquid Chromatographic determination of ascorbic acid and hexavalent chromium in rat lung preparations after treatment with sodium chromate in vitro and in vivo. J Chromatogr 489: 283–290PubMedGoogle Scholar
  33. Suzuki Y, Homma K, Minami M, Yoshikawa H (1984) Distribution of chromium in rats exposed to hexavalent chromium and trivalent chromium aerosols. Ind Health 22: 261–277PubMedGoogle Scholar
  34. Tandon SK (1982) Organ toxicity of chromium in animals. In: Langård S (ed) Biological and environmental aspects of chromium. Elsevier Biomedical Press, Amsterdam, New York, Oxford, pp 209–220Google Scholar
  35. Wiegand HJ, Ottenwälder H, Bolt HM (1984 a) The reduction of chromium(VI) to chromium(III) by glutathione: an intracellular redox pathway in the metabolism of the carcinogen chromate. Toxicology 33: 341–348PubMedGoogle Scholar
  36. Wiegand HJ, Ottenwälder H, Bolt HM (1984 b) Disposition of intratracheally administered chromium(III) and chromium(VI) in rabbits. Toxicol Lett 22: 273–276PubMedGoogle Scholar
  37. Willis RJ, Kratzing CC (1974) Ascorbic acid in rat lung. Biochem Biophys Res Commun 59: 1250–1253PubMedGoogle Scholar
  38. Willis RJ, Kratzing CC (1976) Extracellular ascorbic acid in lung. Biochim Biophys Acta 444: 108–117PubMedGoogle Scholar
  39. Yassi A, Nieboer E (1988) Carcinogenicity of chromium compounds. In: Nriagu JO, Nieboer E (ed) Chromium in the natural and human environments. John Wiley & Sons, New York, Chichester, Brisbane, Toronto, Singapore, pp 443–495Google Scholar

Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • Yasutomo Suzuki
    • 1
  • Kazuo Fukuda
    • 2
  1. 1.Department of Occupational DiseasesNational Institute of Industrial HealthKawasakiJapan
  2. 2.Department of Experimental ToxicologyNational Institute of Industrial HealthKawasakiJapan

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