ANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF DENAVERINE HYDROCHLORIDE IN BULK AND INJECTABLE PHARMACEUTICAL DOSAGE FORM

B. Tharun Kumar*, A. Yasodha and A. Rajamani

ABSTRACT

A new, simple, rapid, precise, accurate and reproducible RP-HPLC method for estimation of Denaverine HCL in bulk form and marketed formulation. Separation of Denaverine HCL was successfully achieved on a Develosil ODS HG-5 RP C18, 5μm, 15cmx4.6mm i.d. column in an isocratic mode of separation utilizing Methanol : Phosphate buffer (0.02M, pH-3.6) in the ratio of 45:55% v/v at a flow rate of 1.0 mL/min and the detection was carried out at 255nm. The method was validated according to ICH guidelines for linearity, sensitivity, accuracy, precision, specificity and robustness. The response was found to be linear in the drug concentration range of 12-28mcg/mL for Denaverine HCL. The correlation coefficient was found to be 0.9995 for Denaverine HCL. The LOD and LOQ for Denaverine HCL were found to be 5.004μg/mL and 15.164μg/mL respectively. The proposed method was found to be good percentage recovery for Denaverine HCL, which indicates that the proposed method is highly accurate. The specificity of the method shows good correlation between retention times of standard solution with the sample solution. Therefore, the proposed method specifically determines the analyte in the sample without interference from excipients of pharmaceutical dosage forms.

Keywords: Denaverine HCL, RP-HPLC, Accuracy, Precision, Robustness, ICH Guidelines.