Component-based biocompatibility and safety evaluation of polysorbate 80

Abstract

Polysorbate 80 is a complicated mixture consisting of a series of esters and etherates synthesized separately by oleic acid and ethylene oxide with a two-core matrix of sorbitan and isosorbide. The extreme complexity of its components makes it difficult to determine its safety. In this study, we provided a quantitative method using UPLC-Q-TOF-MS to identify its components and further explored the side effects of its main constituents using an in vitro hemolysis study, LO2 cells cytotoxicity assay and an in vivo acute toxicity assay to zebrafish. We separated nine major components containing a total of 355 chemicals, confirmed their structures and accurately determined their content. In vitro and in vivo studies revealed that polyoxyethylene sorbitan dioleate (PSD) could induce the highest hemolysis rate, highest cytotoxicity to LO2 cells and highest fatality rate of zebrafish in spite of its relatively low content in polysorbate 80. The results of the exhaustive component-based investigation on polysorbate 80 provide a basis for risk control and safety improvement, and brings new ideas for the re-evaluation and application of any complicated pharmaceutical excipients.