A stability-indicating, ion-pairing, reversed-phase liquid chromatography method for studies of daunorubicin degradation in i.v. infusion fluids

• Published in: Journal of pharmaceutical and biomedical analysis 2013-09, Vol.83, p.164-170
• Main Authors: Respaud, R., Quenum, L., Plichon, C., Tournamille, J.F., Gyan, E., Antier, D., Viaud-Massuard, M.C.
• Format: Article
• Language: English
• Subjects: acetonitrile; acid hydrolysis; aqueous solutions; Chromatography, High Pressure Liquid - methods; Chromatography, Reverse-Phase - methods; Daunorubicin; Daunorubicin - chemistry; Drug Stability; guidelines; high performance liquid chromatography; HPLC; Hydrolysis; Ion-pairing agents; Ions - chemistry; light scattering; oxidation; reversed-phase liquid chromatography; Sensitivity and Specificity; Stability study; Stability-indicating
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2013.05.007

HPLC analysis of a solution of daunorubicin (0.4mg/mL) with different chromatographic conditions: (A) Column: Synergi MAX-RP C12 4μm (150mm×4.6mm I.D.). Isocratic elution: 10mM acid acetic buffer (pH 4.8) and acetonitrile (62:38, v/v). (B) Column: Polar-RP column 4μm (150mm×4.6mm I.D.). Isocratic elution: 6.2mM NFPA in water and acetonitrile (65:35, v/v). (C) Column: Synergi MAX-RP C12 4μm (150mm×4.6mm I.D.). Isocratic elution: 6.2mM NFPA in water and acetonitrile (65:35, v/v). Flow rate: 1mLmin−1; injection volume: 20μL; UV detection at 254nm. •Development of a stability-indicating HPLC method for daunorubicin.•Effect of different stationary and mobile phases on the retention of daunorubicin.•This method could be used for other anthracyclines.•Therapeutic daunorubicin solutions are physically and chemically stable over a period of 14 days. A new stability-indicating method based on high-performance liquid chromatography coupled to ultraviolet and evaporative light scattering detection (HPLC-UV-ELSD) was developed for the quantification of daunorubicin. This is an ion-pairing, reversed-phase method. The column was a Synergi MAX-RP C12 4μm (150mm×4.6mm). The mobile phase was 6.2mM nonafluoropentanoic acid in aqueous solution and acetonitrile under isocratic elution mode. The drug was subjected to oxidation, basic and acid hydrolysis to apply stress conditions. Good resolution was achieved between daunorubicin, related products and all degradation products in an overall analytical run time of approximately 16min with the parent compound daunorubicin eluting at approximately 8min. The method was fully validated according to ICH guidelines and SFSTP protocols in terms of accuracy, precision, specificity and linearity. For daunorubicin, the decision criteria selected consisted of the acceptability limits (±3%) and the proportion of results within the calculated tolerance intervals (95%). In conclusion, the proposed analytical procedures were validated over the selected validation domains daunorubicin (0.25–0.45mg/mL) and shown to provide a very effective method. Physical and chemical stability study was carried out on daunorubicin preparation in our hospital centralized pharmacy unit.