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Updated: Jan 26, 2026
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Quantification of Endothelial Fatty Acid Uptake using Fluorescent Fatty Acid Analogs
Published on: August 15, 2025
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Monitoring polysorbate hydrolysis in biopharmaceuticals using a QC-ready free fatty acid quantification method
Maximilian N Honemann1, Jan Wendler1, Tobias Graf2
1F. Hoffmann-La Roche, Grenzacherstrasse 124, 4070 Basel, Switzerland.
Summary
Polysorbate hydrolysis in biopharmaceuticals causes protein instability and particulates. A new liquid chromatography-mass detection method accurately quantifies free fatty acids, aiding quality control for drug product stability.
Area of Science:
- Biopharmaceutical formulation development
- Analytical chemistry
- Protein stability studies
Background:
- Long-term storage of biopharmaceutical liquid formulations faces challenges due to polysorbate surfactant hydrolysis.
- Polysorbate degradation products can lead to protein instability and the formation of visible/subvisible particulates.
- Current methods for quantifying free fatty acids from polysorbate hydrolysis are indirect, insensitive, and unsuitable for routine quality control.
Purpose of the Study:
- To develop and qualify a novel, label-free analytical method for sensitive quantification of free fatty acids.
- To enable routine quality control testing for polysorbate hydrolysis in biopharmaceutical formulations.
- To assess the real-time storage stability of a formulated monoclonal antibody using the developed method.
Main Methods:
- Development and qualification of a liquid chromatography-single quadrupole mass detector (LC-QDa) based method.
- Utilizing a label-free approach for direct detection and quantification of free fatty acids.
- Application of the method in a case study for real-time storage stability assessment of a monoclonal antibody formulation.
Main Results:
- The developed LC-QDa method is capable of resolving slight changes in the free fatty acid profile.
- The method is reliable and suitable for integration into quality control laboratories.
- The study demonstrated the practical utility of the method in monitoring the storage stability of a biopharmaceutical product.
Conclusions:
- A sensitive and reliable LC-QDa based method for quantifying free fatty acids from polysorbate hydrolysis has been established.
- This method offers a significant improvement over existing techniques for routine quality control in biopharmaceutical development.
- The developed method aids in ensuring the quality and stability of biopharmaceutical liquid formulations.
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