Skip to main content
Log in

High-performance liquid chromatography assay of bleomycin in human plasma and rat hepatocytes in culture

  • Original Articles
  • Bleomycin, HPLC, Rat Hepatocytes, Plasma
  • Published:
Cancer Chemotherapy and Pharmacology Aims and scope Submit manuscript

Summary

A sensitive and rapid linear-gradient, ionpaired, reversed-phase high-performance liquid chromatography technique using fluorescence detection was developed to quantify bleomycin (BLM) metabolites in the plasma of patients undergoing BLM therapy and in rat hepatocytes that had previously been incubated with 5×10−5 m BLM. We could detect about 70 ng/ml using this procedure. BLM metabolites were assayed in the supernatant fractions of precipitated human plasma and in pellets of rat hepatocytes. Metabolite concentrations were below the level of detection in human plasma samples. In hepatocyte pellets, metabolites such as deamido-BLM A2 and deamido-BLM B2 were detected, indicating that isolated rat hepatocytes in culture can metabolize BLM analogues to the corresponding deamido-BLMs. The high-performance liquid chromatography procedure developed during this work can be used to study the metabolism of BLM in cell-culture systems.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Akiyama SI, Kuwano M (1981) Isolation and preliminary characterization of bleomycin-resistant mutants from Chinese hamster ovary cells. J Cell Physiol 107:147

    Google Scholar 

  2. Akiyama SI, Ikezaki K, Kuramochi H, Takahashi K, Kuwano M (1981) Bleomycin-resistant cells contain increased bleomycin-hydrolase activities. Biochem Biophys Res Commun 101:55

    Google Scholar 

  3. Alberts DS, Chen HSG, Mayersohn M, Perrier D, Moon TE, Gross JF (1979) Bleomycin pharmacokinetics in man: II. intracavitary administration. Cancer Chemother Pharmacol 2:127

    Google Scholar 

  4. Aszalos A, Crawford J, Vollmer P, Kantor N, Alexander T (1981) High-performance liquid chromatographic determination of components of bleomycin. J Pharm Sci 70:878

    Google Scholar 

  5. Bedrossian CWM, Luna MA, Mackay B, Lichtiger B (1973) Ultrastructure of bleomycin toxicity. Cancer 32:44

    Google Scholar 

  6. Berry MN, Friend DS (1969) High-yield preparation of isolated rat liver parenchymal cells. A biochemical and fine structure study. J Cell Biol 43:506

    Google Scholar 

  7. Blum RH, Carter SK, Agre K (1973) A clinical review of bleomycin: a new antineoplastic agent. Cancer 31:903

    Google Scholar 

  8. Broughton A, Strong JE (1976) Radioimmunoassay of bleomycin. Cancer Res 36:1418

    Google Scholar 

  9. Comis RL, Kuppinger MS, Ginsberg SJ, Crooke ST, Gilbert R, Auchincloss JH, Prestayko AW (1979) Role of single-breath carbon monoxide-diffusing capacity in monitoring the pulmonary effects of bleomycin in germ cell tumor patients. Cancer Res 39:5076

    Google Scholar 

  10. Crooke ST, Bradner WT (1977) Bleomycin, a review. J Med 7:373

    Google Scholar 

  11. De Lena M, Guzzon A, Monfardini S, Bonadonna G (1972) Clinical radiological and histopathological studies on pulmonary toxicity induced by treatment with bleomycin (NSC-125 066). Cancer Treat Rep 56:343

    Google Scholar 

  12. Henichart JP, Bernier JL, Helbecque N, Houssin R (1985) Is the bithiazole moiety of bleomycin a classical intercalator. Nucleic Acids Res 13:6703

    Google Scholar 

  13. Henichart JP, Bernier JL, Houssin R, Lohez M, Kenani A, Catteau JP (1985) Copper(II)- and iron(II)-complexes of methyl 2-(2-aminoethyl)-aminomethyl-pyridine-6-carboxyl histidinate (AMP His), a peptide mimicking the metal-chelating moiety of bleomycin. An ESR investigation. Biochem Biophys Res Commun 126:1036

    Google Scholar 

  14. Iqbal ZM, Kohn KW, Ewig RAG, Fornace AJ (1976) Single strand scission and repair of DNA in mammalian cells by bleomycin. Cancer Res 36:3834

    Google Scholar 

  15. Israel L, Breau JL, Morere JF, Aquielera J, Khon M (1986) Induction de réponses objectives par une chimiothérapie anticancéreuse fondée sur l'association prolongée de bléomycine et de cisplatine. Presse Med 25:1183

    Google Scholar 

  16. Klett RP, Chovan JP (1985) Modification of a new high-performance liquid chromatography method for bleomycin to separate epi-, iso-, desamido and unmodified analogues. J Chromatogr 337:182

    Google Scholar 

  17. Klett RP, Chovan JP, Raisfeld-Danse IH (1984) Reversed-phase paired-ion high-performance liquid chromatographic method for the separation and quantification of multiple bleomycin congeners. J Chromatogr 310:361

    Google Scholar 

  18. Krack G, Goethals F, Deboyser D, Roberfroid M (1980) Interference of chemicals with glycogen metabolism in isolated hepatocytes. Toxicology 18:213

    Google Scholar 

  19. Krack G, Gravier O, Roberfroid M, Mercier M (1980) Subcellular fractionation of isolated rat hepatocytes. A comparison with liver homogenate. Biochim Biophys Acta 632:619

    Google Scholar 

  20. Lazo JS, Humphreys CJ (1983) Lack of metabolism as the biochemical basis of bleomycin-induced pulmonary toxicity. Proc Natl Acad Sci USA 80:3064

    Google Scholar 

  21. Lazo JS, Boland CJ, Schwartz PE (1982) Bleomycin hydrolase activity and cytotoxicity in human tumors. Cancer Res 42:4026

    Google Scholar 

  22. Lazo JS, Merill WW, Pham ET, Lynch TJ, McCallister JD, Ingbar DH (1984) Bleomycin hydrolase activity in pulmonary cells. J Pharmacol Exp Ther 231:583

    Google Scholar 

  23. Miyaki M, Ono T, Hori S, Umezawa H (1975) Binding of bleomycin to DNA in bleomycin-sensitive and bleomycin-resistant rat ascites hepatoma cells. Cancer Res 35:2015

    Google Scholar 

  24. Muraoka Y (1979) Liquid chromatography of bleomycin. In: Hecht SM (ed) Bleomycin: chemical, biochemical and biological aspects. Springer, New York Berlin Heidelberg, p 92

    Google Scholar 

  25. Paladine W, Cunningham TJ, Sponzo R, Donavan M, Olson K, Horton S (1976) Intracavitary bleomycin in the management of malignant effusion. Cancer 38:1903

    Google Scholar 

  26. Sakai TT (1978) Paired-ion high-performance liquid chromatography of bleomycins. J Chromatogr 161:389

    Google Scholar 

  27. Sebti SM, Lazo JS (1987) Separation of the protective enzyme bleomycin hydrolase from rabbit pulmonary aminopeptidases. Biochemistry 26:432

    Google Scholar 

  28. Seki T, Muraoka Y, Takahashi K, Horinishi H, Umezawa H (1985) Radioimmunoassay of bleomycins. J Antibiot 38:1251

    Google Scholar 

  29. Shiu GK, Goehl TJ (1980) High-performance paired-ion liquid chromatography determination of bleomycin A2 in urine. J Chromatogr 181:127

    Google Scholar 

  30. Shiu GK, Goehl TJ, Pitlick WH (1979) Rapid high-performance liquid chromatographic determination of bleomycin A2 in plasma. J Pharm Sci 68:232

    Google Scholar 

  31. Suzuki H, Nagai K, Yamaki H, Tanaka N, Umezawa H (1969) On the mechanism of action of bleomycin-scission of DNA strands in vitro and in vivo. J Antibiot 22:446

    Google Scholar 

  32. Umezawa H, Maeda K, Takeuchi T, Okami Y (1966) New antibiotics, bleomycins A and B. J Antibiot 19:200

    Google Scholar 

  33. Umezawa H, Suhara Y, Takita T, Maeda K (1966) Purification of bleomycins. J Antibiot 19:210

    Google Scholar 

  34. Umezawa H, Takeuchi T, Hori S, Sawa T, Ishizuka M, Ichikawa T, Komai T (1972) Studies on the mechanism of the anti-tumor effect of bleomycin on squamous cell carcinoma. J Antibiot 25:409

    Google Scholar 

  35. Umezawa H, Hori S, Sawa T, Yoshioka T, Takeuchi T (1974) A bleomycin-inactivating enzyme in mouse liver. J Antibiot 27:419

    Google Scholar 

  36. Yee GC, Crom WR, Brodeur GM, Champion JE, Gutierrez ML, Lee FH, Evans WE (1983) Bleomycin disposition in children with cancer. Clin Pharmacol Ther 33:668

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mahdadi, R., Kenani, A., Pommery, N. et al. High-performance liquid chromatography assay of bleomycin in human plasma and rat hepatocytes in culture. Cancer Chemother. Pharmacol. 28, 22–26 (1991). https://doi.org/10.1007/BF00684951

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00684951

Keywords

Navigation