A fast and validated chromatographic method for simultaneous determination of deferoxamine and D-penicillamine via chelate formation with metal ions in bulk and dosage forms

摘要

Purpose: To develop a chromatographic method for the determination of deferoxamine (DFX) and D-penicillamine (D-PEN) by improving ultra violet (UV)-absorption via complex formation with Fe 2+ and Cu 2+ metal ions. Methods: Chromatographic analysis was performed by Waters RP-HPLC system using a Symmetry? C (18) column with a mobile phase comprising 0.1 % formic acid and methanol (95:5 v/v). For complexation process, drug and metal ion solution were mixed in a ratio of 1:5 and the resulting complex directly analyzed. Validation and system suitability parameters (including chromatographic parameters) were determined. Results: DFX-Fe 2+ and D-PEN-Cu 2+ complexes showed good UV absorption at 260 nm and were easily determined by the newly developed HPLC method. The developed method showed linearity over the concentration range of 8 - 96 μgmL -1 (R 2 0.999 for DFX and 0.99 for D-PEN). Precision and accuracy were also within acceptable limits (100.0 ± 2.0 %). Conclusion: The developed method is robust and validated, and satisfies all the system suitability requirements as per ICH guidelines. DFX injection and D-PEN capsule dosage forms can be successfully analysed with the proposed method. The method is simple, fast and cost-effective for the analysis of D-PEN and DFX individually, or simultaneously in bulk drugs as well as in capsule and parenteral formulations, using UV-detector.