Development and validation of rp-hplc method for the simultaneous estimation of buprenorphine and naloxone in bulk and pharmaceutical dosage form

Abstract

A new, simple, precise, accurate and reproducible RP-HPLC method for Simultaneous estimation of Buprenorphine and Naloxone in bulk and pharmaceutical formulations. SeparationofBuprenorphine and Naloxone wassuccessfullyachievedonaPhenomenex Luna C18 (4.6×250mm, 5µm) particle size or equivalentin an isocratic mode utilizingAcetonitrile: Phosphate Buffer (pH-4.6) (45:55 v/v)ataflowrateof1.0mL/minandeluates wasmonitoredat245nm,witharetentiontimeof2.102and 3.537 minutes for Buprenorphine and Naloxone respectively. The method was validated and the response was found to be linearinthe drug concentration range of 6µg/mLto 14µg/mL for Buprenorphine and 18µg/mLto 42µg/mL for Naloxone. The values of the slopeand the correlation coefficient were found to be 77824 and 0.999 for Buprenorphine and 10515 and 0.999 for Naloxone respectively. The LOD and LOQ for Buprenorphine were found to be 0.6µg/mL and1.8µg/mL respectively. The LOD and LOQ for Naloxone were found to be 0.8µg/mL and 2.4µg/mL respectively. This method was found to be good percentage recovery for Buprenorphine and Naloxone were found to be 100.351 and 100.93 respectively indicates that the proposed method is highly accurate. The specificity of the method shows good correlation between retention times of standard with the sample so, the method specifically determines the analytes in the sample without interference from excipients of tablet dosage forms. The method was extensively validated according to ICH guidelines for Linearity, Range, Accuracy, Precision, Specificity and Robustness.