The Validation of a New Stability-Indicating HPLC Technique Developed for the Quantitative Analysis of Rooperol Tetra-Acetate

Abstract

A high performance liquid chromatographic method was developed for the quantitative analysis of rooperol tetra-acetate in the aqueous receptor phase of in vitro diffusion cells. System validation of the developed HPLC technique was carried out through the determination of the precision, limit of quantitation, sensitivity, linearity, and range of the system. In each of the above tests mentioned, an isocratic mobile phase of acetonitrile/water facilitated the elution of rooperol tetra-acetate dissolved in acetonitrile. Experimental variations were controlled through the use of the internal standard, prazepam. The precision results, expressed as the percent relative standard deviation, ranged from between 1.35% to 4.23%. The results obtained in all other tests performed were acceptable thus verifying that the analytical method used is of good design. Further investigations on rooperol tetra-acetate involved performing stability tests, thus chromatographically characterizing the degradation process of the drug. Heat degradation of rooperol tetra-acetate, when stored in a glass vial and a plastic holder, was assessed over a twelve week period. Data evaluation involved the application of the t-distribution test, used to determine if a significant difference, at a 95% confidence level, exists between the drugs stored under heat and a sample stored at room temperature.