Online SEC/MS Analysis of Proteins on Unix SEC-300: Effect of Mobile Phase Modifier

Online SEC/MS Analysis of Proteins on Unix SEC-300: Effect of Mobile Phase Modifier

Conditions

column Unix SEC-300, 30 cm x 4.6 mm I.D., 1.8 µm
mobile phase [A] modifier (as indicated); [B] 100 mM ammonium acetate, adjusted to pH 7.0 with ammonium hydroxide; (90:10; A:B)
flow rate 0.3 mL/min
pressure 2170 psi (no modifier); 2940 psi (isopropanol); 2600 psi (methanol)
column temp. 35 °C
detector UV, 215 nm
injection 1 µL

Description

Analysis Note This application demonstrates the importance of the inclusion of an organic modifier in online SEC/MS. The organic modifier minimizes any interactions between the analytes and free silanols on the silica stationary phase. Improved efficiency and resolution of the protein analytes were observed when an organic modifier was included in the mobile phase, with methanol yielding the most improvement.
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Application No. ASC140218D

Materials

     
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