Fast and sensitive determination of Polysorbate 80 in solutions containing proteins

A fast and sensitive method has been developed for the specific determination of Polysorbate 80 (Tween 80) in liquid formulations in the presence of proteins and excipients. The quantitative determination is based on a fast liquid chromatographic (HPLC) separation and charged aerosol detection (CAD). The method was validated using a Poroshell 300SB-C18 column packed with 5 μm shell particles (75 mm × 2.1 mm) and acetonitrile–methanol–water–trifluoroacetic acid mobile phase at a flow rate of 0.65 ml/min. The rapid LC–CAD method is suitable for quantifying Polysorbate 80 in the range of 10–60 μg/ml in protein solutions within good manufacturing practices (GMPs) of the pharmaceutical industry.