FAMS-A Targeted Fatty Acid Mass Spectrometry Method for Monitoring Free Fatty Acids from Polysorbate Hydrolysis.

Area of Science:

• Pharmaceutical Science
• Analytical Chemistry
• Biochemistry

Background:

• Polysorbates are essential surfactants for stabilizing protein therapeutics.
• Hydrolytic degradation of polysorbates releases fatty acids, leading to particle formation.
• Quantifying these fatty acids is critical for assessing formulation stability and risk.

Purpose of the Study:

• To provide a detailed protocol for the Fatty Acids by Mass Spectrometry (FAMS) method.
• To enable high-throughput quantification of polysorbate degradation products.
• To ensure method accessibility and successful implementation in diverse laboratories.

Main Methods:

• Development and validation of the Fatty Acids by Mass Spectrometry (FAMS) method.
• Utilized a QDa detector for accessible mass spectrometry analysis.
• Detailed manual and automated sample preparation procedures for high-throughput analysis.
• ICH Q2 (R1) guideline validation for broad applicability.

Main Results:

• The FAMS method accurately quantifies free fatty acids from polysorbate degradation.
• The method is validated for various products, buffers, and protein concentrations.
• End-to-end workflow automation is feasible from sample preparation to data analysis.

Conclusions:

• The FAMS method provides a robust and accessible approach for monitoring polysorbate degradation.
• This protocol facilitates the assessment of particle formation risks in protein formulations.
• Successful implementation is supported by detailed operational guidance and troubleshooting tips.