Oxidation of polysorbates – An underestimated degradation pathway?

Weber, Johanna; Buske, Julia; Mäder, Karsten; Garidel, Patrick; Diederichs, Tim

doi:10.1016/j.ijpx.2023.100202

Cited by 5 publications

(12 citation statements)

References 182 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These observations are in line with general polysorbate raw material storage conditions, in which small (preferential light-protected) containers are used and are covered with an inert gas to limit the oxygen accessibility [ 22 , 60 ]. The presence of proteins would affect the oxidation of polysorbate as well, as elaborately discussed and summarized in Weber et al (2023) [ 60 ]. Radicals formed upon PS oxidation can additionally react with proteins, causing their oxidation.…”

Section: Discussionsupporting

confidence: 59%

“…Additionally, this provides a systematic comparison between the oxidation of PS20 and PS80 for various concentrations. Usually, PS80 is expected to experience faster oxidation based on the high content of unsaturated fatty acids [ 7 , 24 , 31 , 54 , 55 , 56 , 57 ], however, an in-depth comparison between both polysorbates is currently missing in the literature [ 60 ]. FMA measurements were performed to obtain information on PS content.…”

Section: Resultsmentioning

confidence: 99%

“…A number of key characteristics in relation to the oxidative degradation of polysorbate were identified, such as (i) pH shifts in unbuffered solutions [ 9 ], (ii) preferred degradation of higher-order esters [ 25 , 26 , 54 , 55 , 58 ], (iii) dominant degradation of longer fatty acid esters [ 55 , 59 ], (iv) competition with proteins for oxidation [ 27 , 30 , 54 ], and (v) emerging species with lower hydrophobicity for PS80 [ 25 , 26 , 31 , 54 , 55 ]. For a more detailed description and summary, see Weber et al (2023) [ 60 ].…”

Section: Introductionmentioning

confidence: 99%

“…As a result, a larger oxidizability constant of 2.65 was reported for PS80 in comparison to PS20 [ 57 ]. Additionally, more degradation products or oxidation markers were found in PS80 originating from the vicinity of the double bond, such as esters of 9-oxo-C9:0, [ 25 , 55 ] hydroxyl-C18:1 [ 25 , 55 , 56 ], hydroperoxyl-C18:1 [ 25 , 55 ], keto-C18:1 [ 25 , 55 ], epoxy-C18:0 [ 55 , 56 ], 1,9-nonanedioic acid [ 54 ], or 2-decenedioic acid [ 31 ], as summarized in Figure 1 C. Although numerous studies have addressed the oxidation of PS20 and PS80, a systematic and PS concentration-dependent comparison between both polysorbates under pharmaceutically relevant conditions (5 °C without oxidation accelerators) is currently missing in the literature [ 60 ].…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Comparative Stability Study of Polysorbate 20 and Polysorbate 80 Related to Oxidative Degradation

Kozuch,

Weber,

Buske

et al. 2023

Pharmaceutics

Self Cite

View full text Add to dashboard Cite

The surfactants polysorbate 20 (PS20) and polysorbate 80 (PS80) are utilized to stabilize protein drugs. However, concerns have been raised regarding the degradation of PSs in biologics and the potential impact on product quality. Oxidation has been identified as a prevalent degradation mechanism under pharmaceutically relevant conditions. So far, a systematic stability comparison of both PSs under pharmaceutically relevant conditions has not been conducted and little is known about the dependence of oxidation on PS concentration. Here, we conducted a comparative stability study to investigate (i) the different oxidative degradation propensities between PS20 and PS80 and (ii) the impact of PS concentration on oxidative degradation. PS20 and PS80 in concentrations ranging from 0.1 mg⋅mL−1 to raw material were stored at 5, 25, and 40 °C for 48 weeks in acetate buffer pH 5.5 and water, respectively. We observed a temperature-dependent oxidative degradation of the PSs with strong (40 °C), moderate (25 °C), and weak/no degradation (5 °C). Especially at elevated temperatures such as 40 °C, fast oxidative PS degradation processes were detected. In this case study, a stronger degradation and earlier onset of oxidation was observed for PS80 in comparison to PS20, detected via the fluorescence micelle assay. Additionally, degradation was found to be strongly dependent on PS concentration, with significantly less oxidative processes at higher PS concentrations. Iron impurities, oxygen in the vial headspaces, and the pH values of the formulations were identified as the main contributing factors to accelerate PS oxidation.

show abstract

Section: Discussionsupporting

confidence: 59%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Comparative Stability Study of Polysorbate 20 and Polysorbate 80 Related to Oxidative Degradation

Kozuch,

Weber,

Buske

et al. 2023

Pharmaceutics

Self Cite

View full text Add to dashboard Cite

show abstract

“…Oxidative mechanisms for PSs usually start with a PS molecule losing a hydrogen due to UV, temperature or metals producing an alkyl radical. It is rarely acknowledged in the proposed mechanisms that the commercial PS product already contains peroxides and, as such, oxidation can occur without first generating an alkyl radical [ 11 , 16 , 17 ]. Peroxide species have been observed in commercial PS stocks [ 27 ], as well as their components, PEO [ 28 ] and free fatty acids.…”

Section: Resultsmentioning

confidence: 99%

Determining the Oxidation Mechanism through Radical Intermediates in Polysorbates 80 and 20 by Electron Paramagnetic Resonance Spectroscopy

Sutton,

Rustandi

2024

Pharmaceuticals

View full text Add to dashboard Cite

Polysorbates 20 and 80 (PS20 and PS80) are added to many commercial biologic and vaccine pharmaceuticals. It is commonly known that these polysorbates undergo a radical oxidation mechanism; however, the identity of these radical intermediates has not been clearly determined. Furthermore, PS20 and PS80 differ by the presence of a lauric acid instead of an oleic acid, respectively. The oxidation of PS80 is thought to be centered around the double bond of the oleic acid even though PS20 also undergoes oxidation, making the mechanism of oxidation unclear for PS20. Using commercial stocks of PS20 and PS80 alkyl (R•), alkoxyl (C-O•) and peroxyl (C-OO•) radicals were detected by electron paramagnetic resonance spectroscopy likely originating from radical-initiating species already present in the material. When dissolved in water, the peroxyl radicals (C-OO•) originally in the stocks were not detected but poly(ethylene oxide) radicals were. An oxidative pathway for polysorbates was suggested based on the radical species identified in the polysorbate stock material and solutions.

show abstract

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

Konya,

Ochiai,

Fujiwara

et al. 2024

Rapid Comm Mass Spectrometry

View full text Add to dashboard Cite

RationalePolysorbate 80 (PS80) is an amphipathic, nonionic surfactant that is commonly used to stabilize proteins in biopharmaceutical formulations. PS80 undergoes oxidative and/or enzymatic degradation. However, because PS80 is a complex mixture consisting of many constituents, comprehensive evaluations of its oxidative degradation products are difficult and insufficient.MethodsOur previously reported comprehensive liquid chromatography–tandem mass spectrometry (LC/MS/MS)‐based method for PS80 effectively provides an overall profile of PS80 components under simple LC conditions. In this study, we attempted to shorten the analysis time. Furthermore, PS80 was oxidatively degraded in a solution containing histidine and iron, and the oxidative degradation products were evaluated using a modified LC/MS/MS method. In addition, enzymatically hydrolyzed PS80 samples were analyzed.ResultsWe succeeded in shortening the analysis time from 70 to 20 min while maintaining the resolution of the PS80 components of the same selected reaction monitoring transition. Both the previously reported oxidative degradation products and the newly discovered products were successfully detected, and their composition ratios and changes over time were observed. Changes in the hydrolysates over time are shown in the analysis of the hydrolyzed PS80 samples.ConclusionsThis study clearly showed the presence of changes in PS80 oxidative and/or enzymatic degradation products, including those previously unreported. These results demonstrate that a detailed profiling of PS80 degradation products can be performed using LC/MS/MS, which is less expensive and more generally adopted than high‐resolution MS.

show abstract

Oxidation of polysorbates – An underestimated degradation pathway?

Cited by 5 publications

References 182 publications

Comparative Stability Study of Polysorbate 20 and Polysorbate 80 Related to Oxidative Degradation

Comparative Stability Study of Polysorbate 20 and Polysorbate 80 Related to Oxidative Degradation

Determining the Oxidation Mechanism through Radical Intermediates in Polysorbates 80 and 20 by Electron Paramagnetic Resonance Spectroscopy

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

Contact Info

Product

Resources

About