DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR THE SIMULTANEOUS ESTIMATION OF BUPRENORPHINE AND NALOXONE IN BULK AND PHARMACEUTICAL DOSAGE FORM

A novel, precise, accurate, rapid and cost effective isocratic reverse phase high performance liquid chromatographic (RP-HPLC) method was developed, optimized and validated for the estimation of Buprenorphine and Naloxone in bulk and pharmaceutical dosage forms. The drugs were estimated using Phenomenex Gemini C18 (4.6mm×150mm, 5.0 µm) particle size column. A mobile phase composed of tri ethylamine buffer and methanol in proportion of 32:68 v/v, at a flow rate of 1.0 ml/min was used for the separation. Detection was carried out at 248 nm. The linearity range obtained was 30-70 µg/ml for Buprenorphine and 10-50 µg/ml for Naloxone with retention times (Rt) of 3.297 min and 5.405 min for Buprenorphine and Naloxone respectively. The correlation coefficient values were found to be 0.999 & 0.999. Precession studies showed % RSD values less than 2 % for both the drugs in all the selected concentrations. The percentage recoveries of Buprenorphine and Naloxone were found to be 100.1873% for Buprenorphine and 100.748% for Naloxone respectively. The assay results of Buprenorphine and Naloxone were found to be 99.82%. The limit of detection (LOD) and limit of quantification (LOQ) were 2.6µg/ml and 7.8µg/ml for Buprenorphine and 3.4µg/ml 10.2µg/ml for Naloxone respectively. The proposed method was validated as per the International Conference on Harmonization (ICH) guidelines. The proposed validated method was successfully used for the quantitative analysis of commercially available dosage form.

Keywords: Buprenorphine and Naloxone, RP-HPLC, ICH Guidelines, Validation.