A combined qualitative and quantitative method development and validation of vancomycin hydrochloride injection formulation by HPLC and UV involving quality by design.

A selective, specific, precise, linear, accurate and robust analytical method was developed and validated for the assay of vancomycin HCl in vancomycin hydrochloride injection. Comparative UV spectrophotometric and reverse-phase HPLC were used to develop the quantitative determination. Acetonitrile and pH 2.2 phosphate buffer in the ratio 20:80 v/v were used as the mobile phase, and a flow rate of 1.0 ml/min with a 20 min run time. The detection was carried out at 235 nm with a Nucleosil C (250 × 4.6 mm) 10 μm column, and the ambient column temperature was maintained. The method uses a 20 μl injection volume and diluent as a blank solution in this connection. The method was validated as per the current regulatory guidelines. The linearity of this method was found to be linear in the range of 50-150% of the working concentration, and the correlation coefficient was >0.999. The method's accuracy was within the acceptable range, which was 98.1-101.5%. The method's precision was within an acceptable range of about 0.32% RSD. The analytical solution was stable for up to 48 h at room temperature. The method's robustness was proved by utilizing quality design tools. Stress studies demonstrated the method's stability-indicating nature.