Polysorbate 80, as pharmaceutical excipient and virus inactivating agent, is commonly used in the protein pharmaceutical industry. In this study, a method has been developed for the determination of low concentration of Polysorbate 80 in the presence of high concentration proteins (≤100 mg/ml) and excipients. This colorimetric method is based on the interaction of Polysorbate 80 and Coomassie brilliant blue, and suitable for quantitation of Polysorbate 80 in the range of 10-100 μg/ml. Dozens or hundreds of samples can be quantified simultaneously by using microplate. Besides Polysorbate 80, this method can also be used to determine other types of surfactants in protein solutions, such as Polysorbate 20, Triton X-100, NP40, SDS, Benzalkonium chloride/bromide and PEG4000.
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