Analysis of polysorbates in biotherapeutic products using two-dimensional HPLC coupled with mass spectrometer

The trap column, Shim-pack MAYI-ODS, successfully separated the analytes from the IgG. Polysorbates are complex mixture of polymeric compounds, so many peaks appeared on the total ion chromatogram. Using selected ion monitoring mode simplified the chromatogram. The first peak on the SIM chromatogram was used for quantitation. Recovery and reproducibility (n=6) were checked using the two model samples. Recovery for polysorbate 20 and 80 was 100% and 101%, respectively. Relative standard deviation of retention time was 0.093% and 0.057% and that of area was 1.34% and 1.53%, respectively. Linearity was also checked using standards of polysorbates ranging 0.01-0.2mg/mL. Coefficient of determination was 0.998 for polysorbate 20 and 0.999 for polysorbate 80.