Spectrofluorimetric Determination of Doxorubicin in Spiked Serum and Urine Samples


Department of Chemistry, Faculty of Science, Tabriz Branch, Islamic Azad University, Tabriz, Iran


.A simple spectrofluorimetric method is described for the determination of doxorubicin (DXR) based on its quenching effect on the fluorescence intensity of Tb3+- deferasirox (DFX) complex as a fluorescent probe. The excitation and emission wavelengths were 328 and 545 nm, respectively. The effects of pH, time, order of addition of reagents, concentrations of Tb3+ and DFX and the buffer volume on the quenched fluorescence intensity were investigated and optimized. In the optimum conditions, the decrease of the fluorescence intensity of the system showed a good linear relationship with the concentration of DXR in the range of 20-1000 μg L-1, with a correlation coefficient 0.998. The detection limit (3s) was 6.1 μg L-1 and the relative standard deviation for four replicate determinations of different concentrations of DXR was in the range of 1.7–4.4%. The procedure was successfully applied to the determination of doxorubicin in urine and serum samples


  1. DeVita V.T Rosenberg S.A., Hellman S., Cancer principles and practice of oncology, Lippincott, Williams & Wilkins: Philadelphia, 2001.
  2. Takimoto C.H., DeVita V.T., Hellman S., Rosenberg S.A., Cancer: principles & practice of oncology, 7th ed., Lippincott, Williams & Wilkins: Philadelphia, 2005.
  3. Gregoriadis G., Liposomes as Drug Carriers: Recent Trends and Progress, John Wiley and Sons: New York, 1988.
  4. Sottani C., Poggi G., Melchiorre F., Montagna B., Minoiaa C., 2013, Simultaneous measurement of doxorubicin and reduced metabolite doxorubicinol
  5. byUHPLCââ‚‌“MS/MS in human plasma of HCC patients treated with TACE. J Chromatogr B. 915, 71-78.
  6. Mazuel C., Grove J., Gerin G., Keenan K. P., 2003. HPLC-MS/MS determination of a peptide conjugate prodrug of doxorubicin, and its active metabolites, leucine-doxorubicin and doxorubicin, in dog and rat plasma. J Pharm Biomed Anal. 33, 1093-1102.
  7. Ahmed S., Kishikawa N., Ohyama K., Wada M., Nakashima K., Kuroda N., 2009. Selective determination of doxorubicin and doxorubicinol in rat plasma by HPLC with photosensitization reaction followed by chemiluminescence detection. Tlanta. 78, 94-100.
  8. Sakai-Kato K Saito E., Ishikura K., Kawanishi T., 2010. Analysis of intracellular doxorubicin and its metabolites by ultra-high-performance liquid chromatography. J Chromatogr B. 878, 1466-1470.
  9. Zhou Q., Chowbay B., 2002. Determination of doxorubicin and its metabolites in rat serum and bile by LC: application to preclinical pharmacokinetic studies. J Pharm Biomed Anal. 30, 1063-1074
  10. Arnold R.D., Slack J.E., Straubinger R. M., 2004. Quantification of Doxorubicin and metabolites in rat plasma and small volume tissue samples by liquid chromatography/electrospray tandem mass spectroscopy. J Chromatorg B. 808, 141-152.
  11. Jelińska A., Zając M., Cielecka-Piontek J., Gląb K., Tomaszewicz B., Krause A.,Oszczapowicz I., Wąsowska M., 2008. Validation of a Stability Indicating LC-UV Method for [(N-Morpholine)methylene] daunorubicin Hydrochloride. Chromatographia. 67(S1), 107-111.
  12. Perez-Ruiz T., Martinez-Lozano C., Sanz A., Bravo E., 2001. Simultaneous determination of doxorubicin, daunorubicin, and idarubicin by capillary electrophoresis with laser-induced fluorescence detection. Electrophoresis. 22, 134ââ‚‌“138.
  13. Whitaker G., Lillquist A., Pasas S.A.,O'Connor R., Regan F., Lunte C.E., Smyth M.R., 2008. CE-LIF method for the separation of anthracyclines: application to protein binding analysis in plasma using ultrafiltration. J Separation Sci. 31, 1828-1833.
  14. Alava-Moreno F., Valencia-González M.J., Díaz-García M.E., 1998. Kalman filtering-aided time-resolved solid-surface room temperature phosphorimetry for simultaneous determination of anthracyclines in solution. Analyst. 123, 691-694.
  15. Trevisan G., Poppi R.J., 2003. Determination of doxorubicin in human plasma by excitationââ‚‌“emission matrix fluorescence and multi-way analysis. Anal Chem Acta. 493, 69-81.
  16. Ting B.P., Zhang J., Gao Z., Ying J.Y., 2009. A DNA biosensor based on the detection of doxorubicin-conjugated Ag nanoparticle labels using solid-state voltammetry. Biosensors and Bioelectronics. 25, 282-287.
  17. Ruiz-Medina A., Llorent-Martinez E.J., Ortega-Barrales P., Cordova MLFD. 2011. Lanthanide-sensitized luminescence as a promising tool in clinical analysis. Appl Spectrosc Rev. 46, 561-580.
  18. Georges J., 1993. Lanthanide-sensitized luminescence and applications to the determination of organic analytes. Analyst. 118, 1481ââ‚‌“1486.
  19. Henry J.B., Toddââ‚‌“Sanfordââ‚‌“Davidson, Clinical Diagnosis and Management by Laboratory Methods. 18th edn. Saunders: Philadelphia, PA. 1990.
  20. McPherson R.A., Pincus M.R., Henry's clinical diagnosis and managementby laboratory methods. Saunders: Philadelphia, PA. 2006.