Determination of the Monoclonal Antibody Tocilizumab by a Validated Micellar Electrokinetic Chromatography Method

Abstract

Tocilizumab is a monoclonal antibody used in the treatment of several inflammatory and autoimmune diseases as well as cancers. Tocilizumab improves clinical outcomes and reduce mortality rates in patients with COVID-19 disease. A novel, simple and reliable method was developed to determine tocilizumab using micellar electrokinetic chromatography (MEKC). Separation of tocilizumab and the internal standard, methotrexate, was achieved with a background electrolyte consisting of phosphoric acid buffer and sodium dodecyl sulfate (SDS) with UV detection at 195 nm. The method was linear in the concentration range from 10 to 250 µg/mL with correlation coefficient greater than 0.995. The method was successfully applied to the analysis of human and rat plasma samples with good recoveries. Sample preparation involved protein precipitation followed by dilution of the supernatant. The intra‐ and inter-day precisions were less than 5%, the accuracy varied from − 2.71 to 3.84%. The proposed method has acceptable analytical performance and could be applied in future clinical and pharmacokinetic studies including anticancer therapy.

Capillary electrophoresis‐mass spectrometry for intact protein analysis: Pharmaceutical and biomedical applications (2018–March 2023)

“…EKC analyses of proteins were recently reviewed by Gao et al [190]. MEKC-UV was evaluated for a mAbs charge heterogeneity study [191,192]. Micelle-forming molecules like SDS are among the strongest inhibitors of ESI efficiency, therefore applying counter-migrating micelles or partial-filling techniques is needed to prevent the micelles (and their monomers) to enter the MS or volatile surfactants, for example, perfluorooctanoic acid, can be applied for direct MEKC-MS coupling [193].…”

Determination of ticagrelol in rat plasma and tablets by micellar electrokinetic chromatography coupled with large volume sample stacking: Application to a pharmacokinetic study

“…Micellar electrokinetic chromatography (MEKC) is a powerful CE separation approach that allows the separation of analytes by the differential partition between the micellar phase and the surrounding electrolyte [23,24]. MEKC is a useful CE mode for the analysis of both neutral and ionic analytes in commercial drug/herbal products and biological samples [25][26][27][28][29], food/beverages, and the environment [30][31][32][33]. However, the sensitivity of the CE technique is limited due to the small injection volume and short pathway for UV detection.…”