A small amount of surfactant may be added to antibody drugs to prevent agglutination and adsorption of IgG, and concentration control of surfactant is important for quality control. In this application, Tween 20 (Polysorbate 20) and Tween 80 (Polysorbate 80) which are surfactants frequently used in antibody drugs, in the presence of IgG were LC/MS analyzed using ODP2 HP-2D, a polymer-based reversed phase chromatography column, without pretreatment. Under alkaline conditions, IgG was eluted at hold-up volume and Tweens were held in the column by reversed phase mode. Therefore, Tweens can be analyzed without being influenced by ion suppression of IgG without pretreatment (deproteinization) by introducing only the eluate into MS after 5 minutes when IgG is completely eluted.
Sample : 2μL
(Blank)Tween 20 47mg/L (H2O)
Tween 20 47mg/L + IgG 1g/L (in 0.3M NaCl aq.)
Sample : 2μL
(Blank)Tween 80 57mg/L (H2O)
Tween 80 57mg/L + IgG 1g/L (in 0.3M NaCl aq.)
Column : Shodex ODP2 HP-2D (2.0mm I.D. x 150mm) Eluent : (A) 0.1% NH3 aq. / (B) CH3CN High pressure linear gradient; (B%) 20%(0 to 5min), 20 to 90%(5 to 19min), 90%(19 to 24min), 90 to 20%(24 to 25min) Flow rate : 0.2mL/min Detector : UV(280nm), ESI-MS (SIM Positive) Column temp. : 40°C