Small amounts of surfactants are sometimes added to antibody drugs to prevent aggregation and/or adsorption of IgG. Control of surfactant concentration is an important quality control requirement.
In this application, we analyzed poloxamer 188 in the presence of IgG using an ODP2 HP-2B, a polymer-base reversed phase chromatography column, and an MS detector. Under an alkaline condition, IgG is not retained and eluted in the holdup volume. By introducing the sample eluted after 3 minutes, after complete elution of IgG, to the MS, poloxamer 188 can be analyzed without deprotonation pretreatment nor having influenced from IgG ion suppression. Poloxamer 188 is a type of multi-component surfactant which has a range of molecular weight distribution. By selectively analyzing one of the components using SIM mode, highly selective and good qualitative analysis can be achieved. The calibration curve was prepared from poloxamer 188 peak area. Good linearity was obtained in the concentration range 10 to 100 ug/mL. The recovery rate of 50 ug/mL injection was 96 %.
Sample : 2 μL
Poloxamer 188 50 μg/mL + Polyclonal IgG (from human serum) 10 mg/mL (in H2O)
Column : Shodex ODP2 HP-2B (2.0 mm I.D. x 50 mm) Eluent : (A); 0.1 % NH3 aq./(B); CH3CN High pressure linear gradient; (B %) 20 % to 90 % (0 to 5 min), 90 % (5 to 6 min), 90 to 20 % (6 to 6.1 min), 20 % (6.1 to 8 min) Flow rate : 0.2 mL/min Detector : UV (280 nm), ESI-MS (SIM Positive) Column temp. : 40 °C