A UPLC method development and validation study of Upadacitinib and its impurities in extended - Release oral tablet dosage forms.

ABSTRACT

OBJECTIVE: The primary objective was to develop a concomitant isocratic ultra-performance liquid chromatographic photo-diode array detection method to estimate Upadacitinib and its process-related impurities impurity-1 and impurity-2. Further validation was conducted and studied for possible degradants under stress environments. MATERIALS AND METHODS: All the chemicals and reagents used were of HPLC (acetonitrile, methanol) and analytical grade (trifluoro acetic acid). The ultra-performance liquid chromatography (Agilent 1290 Infinity II LC system) consists of a quaternary pump, a BEH C18 (50×2.1mm, 1.7µ) column, and photo-diode array detector. The method was developed with acetonitrile methanol 0.1% v/v trifluoro acetic acid (502030 v/v/v) mobile phase at 0.2mL/min flow rate within a run time of 5.5min The detection was carried at 231.2nm. RESULTS: The respective retention times achieved were 2.289min (Upadacitinib), 0.972min (Upadacitinib impurity-1), and 3.508min (Upadacitinib impurity-2). The optimized method was validated further, and the linearity range was best fit at 15.0-180.0µg/mL for Upadacitinib and 1.0-12.0µg/mL for both Upadacitinib impurity-1 and 2 respectively. The detection and quantification limits were 4.50µg/mL, 15.00µg/mL (Upadacitinib) and 0.30µg/mL, 1.0µg/mL (Upadacitinib impurity-1 and 2). CONCLUSION: A fast, isocratic, specific, and reproducible ultra-performance liquid chromatographic method was developed and validated for various parameters according to the ICH Q2 (R1) guidelines studies. Stress studies were conducted exposing the sample dilution to various treatments (acid, alkali, peroxide, HPLC water, heat, and UV light). The degradants were well-separated apart from the peaks of the active substance. The stability indicating nature was observed during the degradation. The optimized method can be applied for the separation and estimation of Upadacitinib and its process-related impurities in pharma sector in tablet dosage forms.