Analytical Methods of Estimation of Oxetacaine and its quality determinations in different Pharmaceutical Formulations by Reverse Phase High Performance Liquid Chromatography (RP-HPLC)

Abstract

Objective: The aim of the current research was to design and develop an analytical method for the estimation of Oxetacaine and its quality control determinations of different formulation by reverse phase high performance liquid chromatography (RP-HPLC). Method: In Method I the Agilent HPLC used for the Estimation purpose. C-18 column (250 X 4.6mm i.e., 1µ) as stationary phase and 0.05% formic acid buffer: Methanol in the ratio 30:70 % v/v as mobile phase, with a flow rate of 1.0ml/min. Diluents of Methanol: water 90:10 ratio used for the study with ambient Temp conditions. In Method II the Agilent HPLC used for the Estimation purpose .C8 Column ((250 X 4.6mm i.e., 5µ) as stationary phase and ammonium phosphate buffer pH 4.8: Acetonitrile in the ratio 25:75 % v/v as mobile phase, with a flow rate of 0.9ml/min. Diluents of Acetonitryl: water 50:50 ratios used for the study with ambient Temp conditions. Result: Method 1: Oxetacaine standard (86µg/ml concentration) peak was eluted at average of 5.629min at 3 runs with Asymmetry of 1.29813 and Peak area of 23237896 with the conditions of %RSD of max2. With qualifying of system suitability, accuracy and reproducibility. Some of Marketed suspension formulations were operated under the same instrument conditions and the results were compared with standard Oxetacaine to ensure the percentage purity of Marketed suspensions. Method 2: Oxetacaine standard (196µg/ml concentration) peak was eluted at average of 4.884 min at 3 runs with Asymmetryof1.17244 and Peak area of 360543170 with the conditions of %RSD of max 2. With qualifying of system suitability, accuracy and reproducibility. Some of Marketed suspension formulations were operated under the same instrument conditions and the results were compared with standard Oxetacaine to ensure the percentage purity of Marketed suspensions. Conclusion: The two new method of estimation of oxetacaine like using Method 1: C18 column, 0.05% formic acid buffer: Methanol in the ratio 30:70 % v/v as mobile phase @214nm and Method 2: C8 Column, ammonium phosphate buffer pH 4.8: Acetonitrile in the ratio 25:75 % v/v as mobile phase @214 achieved with simple, precise, accurate and sensitive. In addition the quality control estimating of some of random of F1, F2, F3, F4, F5, and F6 marketed oxetacaine suspensions were analysed using this Two Optimized methods. Out of 6 formulations, 2 formulations F3, F6 determined as substandard quality on the both the methods. Hence proving these two new methods optimal for the further monograph inclusions as well as quality control predictions of oxetacaine in API as well as Finished Pharmaceutical Formulations.