Archives of Pharmacal Research

, Volume 32, Issue 4, pp 605–611 | Cite as

A simple HPLC method for doxorubicin in plasma and tissues of nude mice

  • Ahmed M. Al-Abd
  • Nam Ho Kim
  • Soo-Chang Song
  • Seung Jin Lee
  • Hyo-Jeong Kuh
Article

Abstract

Doxorubicin is a cytotoxic anthracycline that has been used for the treatment of several malignancies. Several HPLC methods have been reported for the quantification of doxorubicin in biological samples. Tissue matrix effect and sample size requirements, however, have been remaining issues for simple and easy-to-adapt analytical methods in small animal experiments. The present study established a simple HPLC method for doxorubicin in plasma and tissues (tumor, heart, spleen, liver, gastrointestinal tract, brain, lung, and kidney) of nude mice. Our method required a small sample volume (100 μL plasma and 10 mg tissue), which made it possible to use each blank tissue for calibration curves. The limit of quantification was 25 ng/mL in plasma and 0.1 to 0.4 μg/mg in other tissues with recovery rates ranging from 52.4 to 95.2%. The linearity, accuracy and precision in all tissues, except gastrointestinal tract (GIT), were found to be acceptable in the range of 25–2000 ng/mL plasma and 0.1–4 ng/mg tissue. This method was used successfully to determine the drug concentration in plasma and tissues of human tumor xenograft-bearing nude mice given intratumoral doxorubicin in a polymeric drug delivery system designed for sustained release. In conclusion, the present method may be useful as a simple and easy-to-adapt, yet, sensitive analytical method of doxorubicin for plasma and tissue pharmacokinetic studies in small animals such as nude mice.

Key words

Doxorubicin HPLC Plasma Tissue Nude mice Human tumor xenograft 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Alminana, N., Polo, D., Rodriguez, L., and Reig, F., Biodistribution study of doxorubicin encapsulated in liposomes: Influence of peptide coating and lipid composition. Prep. Biochem. Biotechnol., 34, 77–96 (2004).PubMedCrossRefGoogle Scholar
  2. Alvarez-Cedron, L., Sayalero, M. L., and Lanao, J. M., High-performance liquid chromatographic validated assay of doxorubicin in rat plasma and tissues. J. Chromatogr. B Biomed. Sci. Appl., 721, 271–278 (1999).PubMedCrossRefGoogle Scholar
  3. Arcamone, F., Cassinelli, G., Fantini, G., Grein, A., Orezzi, P., Pol, C., and Spalla, C., Adriamycin, 14-hydroxydaunomycin, a new antitumor antibiotic from S. peucetius var. caesius. Biotechnol. Bioeng., 11, 1101–1110 (1969).PubMedCrossRefGoogle Scholar
  4. Arnold, R. D., Slack, J. E., and Straubinger, R. M., Quantification of Doxorubicin and metabolites in rat plasma and small volume tissue samples by liquid chromatography/electrospray tandem mass spectroscopy. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci., 808, 141–152 (2004).PubMedCrossRefGoogle Scholar
  5. Bagalkot, V., Farokhzad, O. C., Langer, R., and Jon, S., An aptamer-doxorubicin physical conjugate as a novel targeted drug-delivery platform. Angew. Chem. Int. Ed. Engl., 45, 8149–8152 (2006).PubMedCrossRefGoogle Scholar
  6. Behnia, K. and Boroujerdi, M., Inhibition of aldo-keto reductases by phenobarbital alters metabolism, pharmacokinetics and toxicity of doxorubicin in rats. J. Pharm. Pharmacol., 51, 1275–1282 (1999).PubMedCrossRefGoogle Scholar
  7. Bibby, D. C., Talmadge, J. E., Dalal, M. K., Kurz, S.G., Chytil, K. M., Barry, S. E., Shand, D. G., and Steiert, M., Pharmacokinetics and biodistribution of RGD-targeted doxorubicin-loaded nanoparticles in tumorbearing mice. Int. J. Pharm., 293, 281–290 (2005).PubMedCrossRefGoogle Scholar
  8. Chin, D. L., Lum, B. L., and Sikic, B. I., Rapid determination of PEGylated liposomal doxorubicin and its major metabolite in human plasma by ultraviolet-visible high-performance liquid chromatography. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci., 779, 259–269 (2002).PubMedCrossRefGoogle Scholar
  9. de Bruijn, P., Verweij, J., Loos, W. J., Kolker, H. J., Planting, A. S, Nooter, K., Stoter, G., and Sparreboom, A., Determination of doxorubicin and doxorubicinol in plasma of cancer patients by high-performance liquid chromatography. Anal. Biochem., 266, 216–221 (1999).PubMedCrossRefGoogle Scholar
  10. Gao, Z. G., Lee, D. H., Kim, D. I., and Bae, Y. H., Doxorubicin loaded pH-sensitive micelle targeting acidic extracellular pH of human ovarian A2780 tumor in mice. J Drug. Target., 13, 391–397 (2005).PubMedCrossRefGoogle Scholar
  11. Gilbert, C. M., McGeary, R. P., Filippich, L. J., Norris, R.L., and Charles, B. G., Simultaneous liquid chromatographic determination of doxorubicin and its major metabolite doxorubicinol in parrot plasma. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci., 826, 273–276 (2005).PubMedCrossRefGoogle Scholar
  12. Hortobagyi, G. N., Anthracyclines in the treatment of cancer. An overview. Drugs, 54Suppl 4, 1–7 (1997).PubMedCrossRefGoogle Scholar
  13. Hu, T., Le, Q., Wu, Z., and Wu, W., Determination of doxorubicin in rabbit ocular tissues and pharmacokinetics after intravitreal injection of a single dose of doxorubicin-loaded poly-beta-hydroxybutyrate microspheres. J. Pharm. Biomed. Anal., 43, 263–269 (2007).PubMedCrossRefGoogle Scholar
  14. Kang, G. D., Cheon, S. H., Khang, G., and Song, S. C., Thermosensitive poly(organophosphazene) hydrogels for a controlled drug delivery. Eur. J. Pharm. Biopharm., 63, 340–346 (2006a).PubMedCrossRefGoogle Scholar
  15. Kang, G. D., Cheon, S. H., and Song, S. C., Controlled release of doxorubicin from thermosensitive poly(organophosphazene) hydrogels. Int. J. Pharm., 319, 29–36 (2006b).PubMedCrossRefGoogle Scholar
  16. Kummerle, A., Krueger, T., Dusmet, M., Vallet, C., Pan, Y., Ris, H. B., and Decosterd, L. A., A validated assay for measuring doxorubicin in biological fluids and tissues in an isolated lung perfusion model: matrix effect and heparin interference strongly influence doxorubicin measurements. J. Pharm. Biomed. Anal., 33, 475–494 (2003).PubMedCrossRefGoogle Scholar
  17. Licata, S., Saponiero, A., Mordente, A., and Minotti, G., Doxorubicin metabolism and toxicity in human myocardium: role of cytoplasmic deglycosidation and carbonyl reduction. Chem. Res. Toxicol., 13, 414–420 (2000).PubMedCrossRefGoogle Scholar
  18. Loadman, P. M. and Calabrese, C. R., Separation methods for anthraquinone related anti-cancer drugs. J. Chromatogr. B Biomed. Sci. Appl., 764, 193–206 (2001).PubMedCrossRefGoogle Scholar
  19. Lown, J. W., Anthracycline and anthraquinone anticancer agents: current status and recent developments. Pharmacol. Ther., 60, 185–214 (1993).PubMedCrossRefGoogle Scholar
  20. Ricciarello, R., Pichini, S., Pacifici, R., Altieri, I., Pellegrini, M., Fattorossi, A., and Zuccaro, P., Simultaneous determination of epirubicin, doxorubicin and their principal metabolites in human plasma by high-performance liquid chromatography and electrochemical detection. J. Chromatogr. B Biomed. Sci. Appl., 707, 219–225 (1998).PubMedCrossRefGoogle Scholar
  21. Tassin, J. P., Dubois, J., Atassi, G., and Hanocq, M., Simultaneous determination of cytotoxic (adriamycin, vincristine) and modulator of resistance (verapamil, S 9788) drugs in human cells by high-performance liquid chromatography and ultraviolet detection. J. Chromatogr. B Biomed. Sci. Appl., 691, 449–456 (1997).PubMedCrossRefGoogle Scholar
  22. van Asperen, J., van Tellingen, O., and Beijnen, J. H., Determination of doxorubicin and metabolites in murine specimens by high-performance liquid chromatography. J. Chromatogr. B Biomed. Sci. Appl., 712, 129–143 (1998).PubMedCrossRefGoogle Scholar
  23. Veronese, F. M., Schiavon, O., Pasut, G., Mendichi, R., Andersson, L., Tsirk, A., Ford, J., Wu, G., Kneller, S., Davies, J., and Duncan, R., PEG-doxorubicin conjugates: influence of polymer structure on drug release, in vitro cytotoxicity, biodistribution, and antitumor activity. Bioconjug. Chem., 16, 775–784 (2005).PubMedCrossRefGoogle Scholar
  24. Weinberg, B. D., Ai, H., Blanco, E., Anderson, J. M., and Gao, J., Antitumor efficacy and local distribution of doxorubicin via intratumoral delivery from polymer millirods. J. Biomed. Mater. Res. A., 81, 161–170 (2007).PubMedGoogle Scholar

Copyright information

© The Pharmaceutical Society of Korea 2009

Authors and Affiliations

  • Ahmed M. Al-Abd
    • 1
  • Nam Ho Kim
    • 1
  • Soo-Chang Song
    • 2
  • Seung Jin Lee
    • 3
  • Hyo-Jeong Kuh
    • 1
    • 4
  1. 1.Department of Biomedical Sciences, College of MedicineThe Catholic University of KoreaSeoulKorea
  2. 2.Division of Life ScienceKorea Institute of Science & TechnologySeoulKorea
  3. 3.Department of PharmacyEwha Womans UniveritySeoulKorea
  4. 4.College of MedicineThe Catholic University of KoreaSeoulKorea

Personalised recommendations