ABSTRACT

An easy, quick, precise, active and reproducible LC-MS/MS bio analytical Method was developed in rat plasma for the estimation of Reslizumab using Avelumab as internal standard. This article summarizes the recent progress on bioanalytical LC-MS/MS method developed by using Inertsil ODS column (250x4.6 mm, 5?) with organic mobile phase of 5mM Ammonium Formate: Methanol (60:40) of pH-5.5 with flow rate of 1ml/min and the run time was 7min with detected wavelength of 225nm by using avelumab as an internal standard. The Retention time of Reslizumab and internal standard of Avelumab was found to be 2.281min and 4.436min. The analyte was analyzed by mass spectrometry in the multiple reaction monitoring mode. A Turbo-Ion spray source was interfaced between the HPLC and triple quadrupole mass spectrometer (MDS Sciex API 4000). Where the acquired The MRM transitions were m/z 476.2 ? 134.1masses for Reslizumab and Avelumab was m/z 313.3 ? 245.1 (ISTD) were used for quantification of an analyte and its IS. The calibration curve was linear in the range of 5-100 ng/ml for Reslizumab. The method was validated in terms of accuracy, precision, selectivity, recovery, freeze-thaw stability, bench-top stability, stock solution stability and re-injection reproducibility. The mean recoveryfor drug was obtained 99.8%, where as the mean recovery of IS was 99.4%. The %RSD value at higher concentration and lower concentration in all stability experiments was within 15%. This method is free from ion suppression, ion enhancement and any type of abnormal ionization. The method denotes all the parameters of system suitability, specificity, linearity and accuracy are in good agreement with USFDA guidelines and applied effectively for the investigation of pharmacokinetic studies in rat plasma for the determination of reslizumab in bulk and formulation using Avelumab as internal standard. Keywords: Reslizumab, Avelumab (IS), LC-MS/MS, Validation, Rat plasma
Publication date: 01/04/2024

https://ijbpas.com/pdf/2024/April/MS_IJBPAS_2024_7925.pdf

Download PDF
https://doi.org/10.31032/IJBPAS/2024/13.4.7925