Development of Fast, Precise and Selective RP-HPLC Methods for Identification of Possible Degradation Products of Ivermectin in Isolated Rat Hepatocytes and in Different pH Media

Abstract:

Aim: A new stability indicating RP-HPLC based methods were developed to identify the possible degradation products of Ivermectin in isolated rat hepatocytes and in different pH media using a C₁₈ RP column. Complete validation, including linearity, accuracy, recovery, precision, robustness, stability, and peak purity, was performed. Materials and Methods: HPLC system of UltiMateDionex 3000 DAD with LiChrosorb C₁₈ Column were used in this study. During the investigation, for the removal of interfering fraction of isolated rat hepatocytes a purification procedure, consisting of protein precipitation followed by double filtration was developed. Results: Method I was applied effectively and the results indicated two major hepatic metabolites of Ivermectin. Method II was developed to monitor the chemical stability in close to physiological conditions. The appearance of a new peak and the proportional decrease in the sample concentration indicate a liability in buffer with pH12. Conclusion: The obtained results demonstrated that the degradation products of Ivermectin in hepatic cells and in alkali media are different.

Key words: Ivermectin, In vitro, Metabolism, Chemical stability, RP-HPLC.