<span lang="EN-AU">Development and ICH Validation of a Rapid Reversed Phase-HPLC-DAD Method for Tetracosactide Quantification with Sterility Test Suitability

Tetracosactide, or tetracosactrin, a synthetic analog of ACTH, is critical for assessing adrenal cortex function. Tetracosactide can cause the adrenal gland to release more corticosteroids, including cortisol. The importance of tetracosactide could be extended to study osteoarthrosis, juvenile and adult rheumatoid arthritis, ulcerative colitis, and Crohn's disease. So, it is important to get an easy, rapid, sensitive, traceable, and robust analysis method in the quality control labs to detect tetracosactide. Most existing tetracosactide analytical methods explore the analysis from a single perspective and fail to implement the majority of criteria for validating an analytical method in accordance with established guidelines. Alternatively, they rely on costly techniques, are time-consuming, or require specialized reagents. Therefore, the present study investigates and develops an alternative analytical method that economizes on most of the drawbacks inherent in other analytical approaches. The method used HPLC-DAD in the presence of the BDS Hypersil C۱۸ column with a mobile phase of ACN and ۰.۵% w/v ammonium sulfate buffer in the ratio of ۱:۴, detected at ۲۸۰ nm. Method validation was performed in accordance with ICH Q۲(R۱) guidelines, encompassing system suitability testing for precision, repeatability, and accuracy. The method achieved a short retention time of ۴.۵ minutes, with LOD and LOQ of ۴.۲۶ µg/mL and ۱۲.۹۲ µg/mL, respectively. Recovery ranged from ۹۹.۵۲% to ۱۰۰.۲۷%, and linearity was confirmed with a regression coefficient of ۰.۹۹۹۹. The present method provides a reliable and efficient solution for tetracosactide analysis, making it ideal for pharmaceutical quality control and adrenal diagnostics. Additionally, the method was complemented with a validated sterility testing protocol using the direct inoculation method, confirming the absence of antimicrobial interference in product testing.

کلیدواژه ها:

HPLC-DAD ، Reversed-Phase HPLC ، Method validation ، Tetracosactide ، Cosyntropin ، forced degradation ، Sterility test suitability