European Journal of Biomedical and Pharmaceutical Sciences

DEVELOPMENT AND VALIDATION OF STABILITY INDICATING UPLC-ELSD DETECTION METHOD FOR THE DETERMINATION OF OBETICHOLIC ACID IN BULK AND FINISHED FORMULATIONS

Syed Mastan Ali* and D. Ramachandran

ABSTRACT

A simple, sensitive and stability indicating reverse phase ultra-performance liquid chromatography with ELSD detection method (RP-UPLC) was developed and validated for the estimation of Obeticholic acid in bulk and finished dosage forms. The drug was subjected to various stress Conditions such as hydrolysis, oxidation, photolytic and thermal degradations to investigate the stability indicating ability of the method. Compound is highly stable against stress conditions. Obeticholic acid is an UV inactive compound having lower UV absorbance and so a ELSD coupled with UPLC system was used for better sensitivity. Efficient chromatographic separation was achieved by using Acquity; UPLC, BEH; C-18; 100 x 2.1mm; 1.7 μm column with the mobile phase consisting of 0.05% Trifluoro acetic acid in water and 0.05% Trifluoro acetic acid in acetonitrile in a gradient elution mode within a short run time of 6.0 minutes at a flow rate of 0.3 ml/min. The developed method was validated as per the current ICH quality guidelines with respect to specificity, precision, accuracy, linearity, robustness and solution suitability. The average recovery values of Obeticholic acid were found to be in the range of 99.99-101.41 %. The developed method was linear with the correlation value of 0.9994 for Obeticholic acid. The repeatability and intermediate precision expressed by RSD were less than 2.0% for Obeticholic acid. The test solution was found to be stable in diluent for 48 h when stored at room temperature. The developed UPLC method is superior in technology against conventional HPLC with respect to speed, resolution, solvent consumption and cost of analysis. This method is compatible to LCMS analysis which enables to identify the unknown impurities or the degradants formed in the process.

Keywords: Obeticholic acid, ELSD (Evaporative Light Scattering Detector), stability indicating UPLC method, Assay validation.