P. Swathanthra Babu*, S. Bhayalata, ChaitanyaBangari, Dr.M.B.Venkatapathi Raju
A novel method were recognized for simultaneous estimation of Kanamycin and Cephalexin by RP-HPLC method. The chromatographic conditions were effectively developed for the separation of Kanamycin and Cephalexin by by means of SYMMETRYC18 column (4.6×150mm)5µ, flow rate were1ml/min, mobile phase ratio were (70:30 v/v)methanol: phosphate buffer(KH2PO4and K2HPO4) pH 3 ( pH were accustomed with orthophosphoric acid),detection wavelength were258nm. The instrument utilizing was WATERS HPLC Auto Sampler, Separation module 2695, photo diode array detector 996, Empower-software version-2. The retention times were found to be 2.403 mins and 3.907mins. The % purity of Kanamycin and Cephalexin were establish to be 100.27% and 99.87% respectively. The system suitability parameters for Kanamycin and Cephalexin such as theoretical plates and tailing factor were found to be 2294, 1.27 and 4891 and 1.03, the resolution were found to be 8.67. The analytical method were validated according to ICH guidelines (ICH, Q2 (R1)). The linearity study n Kanamycin and Cephalexin were found in concentration range of 50µg-250µg and 5µg-50µg and correlation coefficient (r2) were found to be 0.999 and 0.999, % recovery were found to be 99.56% and 99.48%, %RSD for repeatability were 0.27 and 0.40, % RSD for intermediate precision were 0.27 and 0.94 respectively. The precision study were precise, robust, and repeatable.LOD value were 2.17 and 6.60, and LOQ value were 0.032 and 0.1125 respectively. Keywords: Kanamycin , Cephalexin , precise, robust, and repeatable.