STABILITY INDICATING RP-HPLC METHOD DEVELOPMENT AND VALIDATION OF ABROCITINIB IN BULK AND TABLET DOSAGE FORM

*Ms. Manasi S. Wagh, Mr. Rajesh G. Jadhao, Dr. Parag R. Patil

ABSTRACT

A simple, precise, accurate and stability-indicating RP-HPLC method was developed and validated for the estimation of Abrocitinib in bulk and pharmaceutical dosage form. The analysis of Abrocitinib in methanol using UV spectrophotometric analysis yielded a well-defined absorption maximum (λ max) which showed that it could be detected analytically. The sharp and clear peak indicated the purity of the analyte and the absence of interference. The chromatographic conditions were optimized based on systematic trials on varying mobile phase compositions. Trial-07 was found to be optimized performing trial in terms of chromatographic performance as it had a retention time of about 4.62 minutes, good peak symmetry (0.92), and acceptable theoretical plate count, allowing good separation and reproducibility. The method had a high linearity in 10-50 μg/mL concentration range with a regression equation of Y = 98.85x + 7.317 and correlation coefficient (R 2) of 0.999 indicating a good linear relationship between concentration and peak area. Recovery studies at 80%, 100% and 120% levels confirmed the accuracy of the method with mean recoveries of 99.74%, 100.69% and 99.34% at each level. The values of the %RSD were below 0.5% indicating good accuracy and no excipients interference. Intraday and interday precision studies demonstrated a very low level of the values of %RSD, below 2%, indicating outstanding repeatability and reproducibility of the method. Repeatability studies also showed that there were stable peak areas with a percentage relative standard deviation of 0.08%. The sensitivity of the procedure was confirmed with low LOD (0.1275 μg/mL) and LOQ (0.3864 μg/mL) implying that the procedure can identify and measure the drug at very low levels. Ruggedness and robustness experiments indicated that small intentional variations in analytical conditions like mobile phase composition and detection wavelength did not have any significant impact on the chromatographic performance. The values of the %RSD were in the satisfactory range that validated the applicability of the method in different conditions. Marketed formulation assay revealed a close to 100% (mean 100.62%) of label claim, which demonstrated that the method could be used as a routine analysis of quality control. Studies on forced degradation at acidic, alkaline, oxidative, and neutral conditions revealed that Abrocitinib is highly vulnerable to oxidative degradation followed by alkaline degradation, and acidic degradation and stable in neutral conditions. Photolytic studies confirmed that the drug is stable under UV exposure. The degradation products were well resolved from the main drug peak, confirming the stability indicating nature of the method.

Keywords: RP HPLC, Analytical method development, Abrocitinib.