Extensive Characterization of Polysorbate 80 Oxidative Degradation Under Stainless Steel Conditions

Detailed profiling of polysorbate 80 oxidative degradation products and hydrolysates using liquid chromatography–tandem mass spectrometry analysis

“…The main methods used to identify the degradation products in other studies are a full scan [7][8][9][10][11][12] or charged aerosol detection. [13][14][15][16] Since the data obtained by charged aerosol detection do not include mass information, distinguishing PS80 degradation products with different esterified FAs is impossible. In addition, in a full scan, the detection of low-concentration oxidative degradation products is usually difficult because the signals of these products may be obscured by those of components present at high concentrations within the same retention time.…”

“…However, commercially available polysorbate products contain substantial amounts of byproducts, and various analytical methods have been developed for their detection. [4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23] PS80 is known to be oxidatively or enzymatically degraded, and the degradation of PS80 can sometimes affect protein stability. 3,7,8,[24][25][26] Therefore, polysorbate degradation has emerged as a major challenge in the biopharmaceutical community.…”

“…Several studies have reported the stability and degradation products of polysorbate. [7][8][9][10][11][12][13][14][15][16] However, because PS80 is a complex mixture comprising many constituents, comprehensive evaluations of its degradation, especially with respect to oxidative degradation products, are difficult and insufficient. The decrease in PS80 components and the increase in its degradation products were evaluated by total ion chromatography [7][8][9][10][11][12] or charged aerosol detection.…”

“…The decrease in PS80 components and the increase in its degradation products were evaluated by total ion chromatography [7][8][9][10][11][12] or charged aerosol detection. [13][14][15][16] Liu et al reported two oxidative degradation pathways and six degradation products (hydroxy-C18:1, epoxy-C18:0, keto-C18:1, hydroperoxy-C18:1, oxo-C9:0, and 2-decenedioic acid ester) of PS80. 7 The structural formula of each degradation product and the difference in molecular weight from the oleate ester are shown in Figure S1.…”

Cationic polymer precipitation for enhanced impurity removal in downstream processing

“…PS-80 was spiked in Protein A purified samples to compare lipase activities. Previous reports indicate PS-80 degradation rates are elevated at higher temperature (Zheng et al, 2023); thus, these samples were held at 40°C for 14 days. The results of the PS-80 (14 days at 40°C) after full downstream purification steps (with the second polishing chromatography).…”