VALIDATION OF RP-HPLC ANALYTICAL METHOD FOR ESTIMATION OF CARMUSTINE IN BULK AND LYOPHILIZED VIALS

Carmustine is an anti-neoplastic agent. A reverse phase high-performance liquid chromatographic (RP-HPLC) assay method was developed by slightly modifying the USP method and validated for quantitative determination of carmustine in bulk drug and in lyophilized vials. The column utilized for the estimation of assay was 4.6 mm X 15-cm, 5-μm Packing L1 (C18) and the mobile phase employed was a mixture of acetonitrile and water in the ratio of 3:7. The detection was carried out at a wavelength of 200 nm with PDA detector. The flow rate is 1.5 mL per minute at a set temperature of 4-6°C with a total run time of 15 minutes. The linearity range was studied ranging from 25-150% with regression coefficient value of 0.999 and the method precision for estimation of assay was below 1.0% RSD. During the forced degradation studies, the drug substance was found to be degraded in all stress conditions because of the nature of the drug substance and the purity angle was found to be less than the purity threshold in all stress conditions. The degradation products were well separated from the main assay peak. The prepared stock solutions of test and standard were found to be stable up to 24hrs at refrigerated condition. Finally it can be concluded that the validated RP-HPLC method can be successfully used as a quality control tool for the estimation of drug substance in the bulk drug and in lyophilized vials.