DEVELOPMENT AND VALIDATION OF A HPLC-UV METHOD FOR SIMULTANEOUS DETERMINATION OF CEFTRIAXONE AND SULBACTAM IN POWDER FOR INJECTION FORMULATION

A simple, rapid, and sensitive high-performance liquid chromatographic method with UV detection has been developed and validated according to the ICH guidelines for the quantization of Ceftriaxone (CEF) and Sulbactam (SUL) in parenteral preparation. Chromatographic separation was carried out in a Hypersil Gold C8 column (250 mm x 4.6 mm; 5 mu m particle size) of Thermo Scientific with simple mobile phase composition of 10 ml of 40% Tetra Butyl Ammonium Hydroxide(TBAH) in 1000 ml of water ( pH 5.5, maintained by dil Phosphoric acid) and acetonitrile ( 70: 30, v/v) at a flow rate of 2.0 ml min(-1) with injection Volume of 20 mu l where detector was set at 227 nm with a total run time of 10 mins. The method was linear over the concentration range of 40-100, mu g ml(-1) for SUL and 80-200 mu g ml(-1) with a correlation coefficient of 0.999 and 0.999 respectively. Limit of quantifications (LOQ) of 8.5, 14.4 and limit of detections (LOD) 2.8, 4.7 mu g ml(-1) for CEF and SUL respectively. Accuracy and precision values of both within-run and between-run obtained from six different sets of three quality control ( QC) samples analyzed in separate occasions for both the analytes ranged from 98.15% to 99.75% and 0.91% to 1.58%, respectively. Extraction recovery of analytes from 97.57% to 99.03%. The developed and validated method was successfully applied to quantitative determination of CEF and SUL in pharmaceutical formulation.