Analytical Method Development and Validation for the Simultaneous Estimation of Buprenorphine and Naloxone by RP- HPLC Method

Dr. Gampa Vijay Kumar^1*, G. Sravanya^2
1Professor and Head, Dept. of Pharmacy, KGR Institute of Technology and Management, Rampally, Kesara, Rangareddy, Telangana, India.
²KGR Institute of Technology and Management, Rampally, Kesara, Rangareddy, Telangana, India.

A  B  S  T  R A C T
High performance liquid chromatography is at present one of the most sophisticated tool of the analysis. The estimation of Buprenorphine and Naloxone was done by RP-HPLC. The Phosphate buffer was p^H 3.0 and the mobile phase was optimized with consists of Methanol: Phosphate buffer mixed in the ratio of 70:30 % v/ v. Inertsil C₁₈ column   C18 (4.6 x 150mm, 5mm) or equivalent chemically bonded to porous silica particles was used as stationary phase. The detection was carried out using UV detector at 260 nm. The solutions were chromatographed at a constant flow rate of 0.8 ml/min. the linearity range of Buprenorphine and Naloxone were found to be from 100-500 mg/ml of Buprenorphine and 1-5mg/ml of Naloxone . Linear regression coefficient was not more than 0.999.The values of % RSD are less than 2% indicating accuracy and precision of the method. The percentage recovery varies from 98-102% of Buprenorphine and Naloxone. LOD and LOQ were found to be within limit. The results obtained on the validation parameters met ICH and USP requirements .it inferred the method found to be simple, accurate, precise and linear. The method was found to be having suitable application in routine laboratory analysis with high degree of accuracy and precision.
Keywords: Methanol: Phosphate buffer, Inertsil C₁₈ column, Buprenorphine and Naloxone