A Study of Method Development, Validation and Forced Degradation for Quantification of Buprenorphine Hydrochloride in a Microemulsion Formulation

Objective: Development and validated of a simple, selective RP-HPLC method for the determination of buprenorphine hydrochloride in pharmaceutical microemulsion formulation. A forced degradation study of developed formulation was carried out in accordance with International Conference on Harmonization (ICH) guidelines Q1 (R2). Method: The chromatogram was obtained with 10 mmolL−1 potassium phosphate buffer adjusted to pH 6.0 with triethanolamine and acetonitrile (17:83, v/v) as mobile phase, C18 HPLC column (250 × 4.6 mm i.d., 5 µm) kept at 30â—¦C and UV detection at 284 nm. The compound was eluted isocratically at a flow rate of 1.0 mL min−1. Results: The average retention time for buprenorphine was 14.319 min. The method was validated according to the ICH guidelines. The validation characteristics included accuracy, precision, linearity, range, specificity, limit of Quantitation and robustness. The calibration curves were linear (R2 > 0.999) over the concentration range 1.0 – 500.0 µgmL−1 for buprenorphine hydrochloride and recovery study for the compound was above 95 %. No spectral or chromatographic interferences from the microemulsion excipients were found. The drug was found to be labile under oxidative stress condition; whereas stable under all other stress conditions. Conclusion: This method is simple, rapid and suitable for routine quality control analysis.