Stability indicating method development and validation of Beclomethasone dipropionate and Neomycin sulphate Ointment using HPLC with microbial assay

Abstract

A simple, economic, selective, precise and accurate stability-indicating RP-HPLC method for the analysis of “Beclomenthasone Dipropionate (BD) an anti inflammatory in its formulation was developed and validated in the present study. Here Neomycin Sulphate (NS) in the ointment is one of a class of aminoglycoside antibiotic that lack a good UV absorbing chromophore, therefore difficult to determine using RP-HPLC with absorbance detection. So Microbiological assay is the best option for their determination. RP-HPLC for Beclomethasone Dipropionate was performed on a Kromosil C18, 5μ, 15cm x 4.6mm column. The mobile phase consist a mixture of ACN: WFI: Methanol: Glacial Acetic Acid: Triethylamine (55: 40: 5: 1: 0.1 v/v). This was found to give sharp peak of Beclomethasone Dipropionate at a wavelength of 235nm with a flow rate of 1.2ml/min. the linear regression analysis data for the calibration curve showed a good linear relationship with a regression coefficient 0.999. The linear regression equation was Y= 64760x – 1875. Linearity was found in the range of 5-15 μg/ml. the accuracy (%recovery) was found to be in the range of 99.07 – 99.85%. NS was performed on the culture medium of the ( Recoiorery) linearity was found in the range of 5-15 μg/ml. the accuracy (% recovery) was found to be in the range of 99.07 - 99.85%. NS was performed on the culture medium of agar was composed of 20 Ml Grove-Randall’s 1 culture medium (Difco) that was poured into a 100 x 20 mm Petri dish used for inhibit the strain of Staphylococcus aureus. The linear regression analysis data for the calibration curve showed a good linear relationship with a regression coefficient 0.998. The linear regression equation was y = 2.401x – 0.001. Linearity was found in the range of 2.5 – 15 μg/ml. The accuracy (% revcovery ) was found to be in the range of 98.49 – 99.03 %. BD and NS were subjected to stress conditi0on0s including acidic, alkaline, oxidation, photolysis and thermal degradation. BD and NS were more sensitive towards acidic degradation. The method was validated as per ICH guideline. The method was validated for accuracy, precsion, robustness, specificity. The proposed methods can be used successfully for routine analysis of Beclomethasone Dipropionate (BD) by HPLC and Neomycin Sulphate (NS) by Microbiological assay in ointment form.