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Abstract

SIMULTANEOUS ESTIMATION METHOD DEVELOPMENT AND VALIDATION OF ACEBROPHYLLINE AND DOXOFYLLINE IN TABLET DOSAGE FORM BY RP-HPLC METHOD

Shinde Madhuri*, H. S. Bhawar and Prof. G. S. Shinde

ABSTRACT

A simple, accurate, precise and reproducible Reverse Phase High Performance Liquid Chromatography method was developed and validated for simultaneous estimation of Acebrophylline and Doxofyllinein tablet dosage form. Chromatographic separation was achieved by Cosmosil C18 (250 mm x 4.6 ID, Particle size- 5 micron) column and Methanol: 10mM KH2PO4 buffer (70:30)as mobile phase, at a flow rate of 0.8 ml/min (millilitre per minute) using UV detection at 243nm. The retention time for Acebrophylline and Doxofylline were obtained as 4.076min and 5.193 min. respectively. The method has been validated for linearity, accuracy, precision, LOD, and LOQ. Linearity of Acebrophylline and Doxofyllinewere found to be 5- 25μg/ml.(R2=0.999) and 20- 100μg/ml.(R2=0.999) respectively. The accuracy of present method was evaluated at 50%, 100%, 150%. Recovery was found to be in a range from 99.46%-99.98% for Acebrophylline and 99.91%-100.12% for Doxofylline. Intermediate precision studies were carried out and the RSD values were less than 2%. Lower values of LOD (0.150μg/ml) and LOQ (0.455μg/ml) for Acebrophylline and LOD (0.127μg/ml) and LOQ (0.385μg/ml) fordoxofylline indicated good sensitivity of the method. In this study, the optimization of mobile phase, flow rate, injection volume and wavelength were achieved. This demonstrate that the developed method is simple, precise, accurate and robust for simultaneous estimation of Acebrophylline and Doxofyllinein tablet dosage form.

Keywords: Acebrophylline and Doxofylline, RP-HPLC, Validation.


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