Validation of a\nSimple HPLC/UV Method for Assay and\nIn Vitro Release of Glycosaminoglycan from Pharmaceutical Formulations

This study encompasses\nthe validation of a simple, rapid,\nand sensitive\nHPLC/UV method developed in accordance with the guidelines set by\nICH Q2(R2) for obtaining the active pharmaceutical ingredient from\nthe glycosaminoglycan family in topical formulations. Previous methods\nreported for analyzing glycosaminoglycans in semisolid formulations\nare relatively complex and time-consuming, involving extraction, purification,\nand derivatization. This developed analytical method allows for straightforward\nextraction of the active pharmaceutical ingredient from the matrix,\nenabling the direct injection of samples. This method was performed\nand validated for the assay of the pharmaceutical gel and cream formulations\nto investigate the parameters of linearity (<i>r</i> = 0.9997\nfor the gel formulation and <i>r</i> = 0.9993 for the cream\nformulation), precision, accuracy, specificity, and robustness by\nHPLC/UV. Additionally, this method was used to determine the active\ningredient in <i>in vitro</i> release studies. <i>In\nvitro</i> similarity correlation against commercial products\nwas performed according to the Mann–Whitney <i>U</i> statistical test. The similarity results were 96.5–102.7%\nfor the gel formulation and 98.0–106.0% for the cream formulation,\nwhich remained within the limits (75–133.33%) according to\nUSP 1724. This proved that <i>in vitro</i> release profiles\nfor both formulations were like those of the commercial product. In\nlight of the research findings, we believe that the HPLC/UV analysis\npresented can be further enhanced in the future for determining the\nlevels of active ingredients in various pharmaceutical formulations\nor for monitoring the levels of glycosaminoglycans in biological matrixes.