Development of an UPLC-MS/MS Method for the Quantitative Analysis of Upadacitinib in Beagle Dog Plasma and Pharmacokinetics Study

Abstract

Background Upadacitinib, a novel selective Janus kinase 1 (JAK1) inhibitor, has been recently approved by the US FDA for the treatment of adult patients with moderately to severely active rheumatoid arthritis (RA). An ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for the quantitative analysis of upadacitinib in beagle dog plasma was developed and validated. Methods Upadacitinib and fedratinib (internal standard, IS) were extracted with ethyl acetate under alkaline condition and then separated and detected. The chromatographic column was Waters Acquity UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm), the mobile phase was acetonitrile and 0.1% formic acid in water with gradient elution procedure, and the flow rate was 0.40 mL/min. Under the positive ion mode, upadacitinib and IS were monitored by multiple reaction monitoring (MRM) as the following mass transition pairs: m/z 447.00 → 361.94 for upadacitinib and m/z 529.82 → 141.01 for IS. Results In the concentration range of 1–500 ng/mL, upadacitinib had good linearity, and the lower limit of quantification (LLOQ) was 1 ng/mL. The RSD of the intra- and inter-day precision was less than 10.03%, and the RE of accuracy was −3.79% to 2.58%. The extraction recovery of upadacitinib was more than 80%, the matrix effect was around 100%, and upadacitinib was found to be stable. Conclusion The novel optimized UPLC-MS/MS assay was an effective tool for the determination of upadacitinib and had been successfully applied to the pharmacokinetic study of upadacitinib in beagle dogs, and this method would also be used to study DDIs.

Isolation and Quantification of the Hepatoprotective Flavonoids From Scleromitron diffusum (Willd.) R. J. Wang With Bio-Enzymatic Method Against NAFLD by UPLC–MS/MS

“…Therefore, a comprehensive method must be established to quantify the flavonoid content and screen out potential differential components. UPLC-MS/MS is a highly efficient and preferred technique for TCM ( Nandan et al, 2021 ; Verma et al, 2021 ) and pharmacokinetics study ( Chen et al, 2021 ; Zhang Z. et al, 2021 ; Lee et al, 2021 ; Wang et al, 2021 ; Zhang et al, 2022 ). In our study, the application combining UPLC-MS/MS and multivariate statistical methods was used to isolate and quantify flavonoids prepared by different methods, followed by differential components screened.…”

Therapeutic Drug Monitoring for Biologic and Small-Molecule Therapies for Inflammatory Bowel Disease

“…There are multiple methods for therapeutic drug monitoring, including enzyme-linked immunoassays (ELISAs), liquid chromatography and radioimmunoassay, with ELISA being the most common in commercial use [28][29][30][31][32]. The key components of these assays are summarised in Table 1.…”

Quantification of an Anti-Rheumatic Agent: Upadacitinib in Biological Fluid (Plasma) By LC- MS/MS

“…Without the right medical attention, irreversible joint injury and loss can cause serious disability and a lower quality of life. With the development of biological therapeutic techniques including anti-TNF, anti-IL-6, anti-CTLA4 Ig, and anti-CD20 over the past 30 years, the treatment of RA has undergone significant changes [3,4]. Because these medications are more effective, patients have access to a wide range of therapy options [5].…”