Method development and alkali degradation study of doxofylline by rp-hplc and lc-ms/ms

RESULT AND DISCUSSION

A sensitive, selective, precise and accurate high performance liquid chromatographicmethod of analysis of doxofylline in both as bulk drug and in formulation was developed and validated. The mobile phase consisted of acetonitril: 0.05M formic Acid (90: 10v/v); pH-3.0. The detection wavelength was 274nm. This system was found to give the sharp peak for Doxofylline (RT-2.9). The method was validated as per ICH guideline. Stability indicating assay method in which alkali stress condition was used for quantitative estimation of doxofylline in tablet formulation and identification of alkali degradation product The separation of drug from its degradation product were optimized by varying the ratio &/or nature of organic modifier. Finally method was developed using same

mobile phase composed of acetonitrile: formic Acid (90: 10); pH-3.0, in that both drug and degradation product showing good elution RT-2.9 (Doxofylline) and RT-5.1 (Alkali

degradation product) and m/e-262. The acid degradation product and pure drug were identified by LC-MS/MS in order to establish acid degradation pathway(fig.-04, 05and

06).