An antibody-drug conjugate (Antibody-Drug Conjugate; ADC) is a combination of an antibody drug and small molecule drugs with linkers. It is attracting attention as a next-generation drug that combines the advantages of low molecular weight drugs and antibody drugs. A small amount of surfactant is often  added to ADC to prevent its aggregation and adsorption, and its concentration control is important for quality control.
Using the column ODP2 HP-2B as polymer-based reverse phase chromatography,the polysorbate 20 formulated in a commercially available ADC was measured by LC / MS. Commercially available ADCs are dissolved in water and then used for measurement. Under alkaline conditions, the ADCs are not retained on the column and are collectively eluted in the holdup volume, in other hands, Polysorbate 20 is retained on the column in reverse phase mode. Therefore, even without pretreatment (deproteinization)
By introducing only the eluate after 1.5 minutes of complete elution of ADC into MS, analysis of polysorbate 20 is possible without the influence of ion suppression by ADC.
Since polysorbate 20 is a multicomponent surfactant, although it is detected as a broad peak in the total ion chromatogram in scan measurement by detecting one of the components in SIM mode, quantitative analysis can be performed well.


Sample : 2 μL
Antibody-drug conjugate formulation
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 10 min) 
Flow rate    : 0.2 mL/min
Detector     : UV (280 nm), ESI-MS (Scan & SIM Positive) 
Column temp. : 40 °C