Validation of a Simple HPLC/UV Method for Assay and In Vitro Release of Glycosaminoglycan from Pharmaceutical Formulations.

Abstract

This study encompasses the validation of a simple, rapid, and sensitive HPLC/UV method developed in accordance with the guidelines set by ICH Q2(R2) for obtaining the active pharmaceutical ingredient from the glycosaminoglycan family in topical formulations. Previous methods reported for analyzing glycosaminoglycans in semisolid formulations are relatively complex and time-consuming, involving extraction, purification, and derivatization. This developed analytical method allows for straightforward extraction of the active pharmaceutical ingredient from the matrix, enabling the direct injection of samples. This method was performed and validated for the assay of the pharmaceutical gel and cream formulations to investigate the parameters of linearity (r = 0.9997 for the gel formulation and r = 0.9993 for the cream formulation), precision, accuracy, specificity, and robustness by HPLC/UV. Additionally, this method was used to determine the active ingredient in in vitro release studies. In vitro similarity correlation against commercial products was performed according to the Mann-Whitney U statistical test. The similarity results were 96.5-102.7% for the gel formulation and 98.0-106.0% for the cream formulation, which remained within the limits (75-133.33%) according to USP 1724. This proved that in vitro release profiles for both formulations were like those of the commercial product. In light of the research findings, we believe that the HPLC/UV analysis presented can be further enhanced in the future for determining the levels of active ingredients in various pharmaceutical formulations or for monitoring the levels of glycosaminoglycans in biological matrixes.