Why analyze polysorbate 20/80 in biopharmaceutical products using HPLC-MS?
Polysorbate 20 and polysorbate 80 are essential excipients in biopharmaceutical formulations. Their main role is to ensure protein stability by reducing the risk of denaturation and aggregation. However, their degradation, caused by oxidative, hydrolytic, or enzymatic processes, can affect the efficacy of drugs and even compromise their safety.
Faced with these challenges, the HPLC-MS (High Performance Liquid Chromatography coupled with Mass Spectrometry) method is a fast and effective approach for accurately analyzing and quantifying polysorbates 20/80 in complex environments such as protein solutions.
An effective "one-pot" analysis method
The concept behind this method, which refers to a process whereby all the necessary steps of an analysis or synthesis are carried out in a single container, is based on two principles: the hydrolysis of esters of polysorbate fatty acids and the rapid methylation of free fatty acids to improve their detection. Thanks to a “one-pot” process, the sample preparation step is simplified, facilitating its integration into formulation, development, and quality control (QC) environments.
The analytical approach is as follows:
- Chromatographic separation: using a high-performance C8 HPLC column.
- High-sensitivity MS detection: for accurate identification of methylated fatty acids present in polysorbates.
The method demonstrates remarkable sensitivity, achieving detection levels below ppm. It thus allows precise quantification of polysorbate constituents, even in complex matrices.
The advantages of the HPLC-MS method
Increased sensitivity and specificity
Unlike traditional approaches such as ELSD or CAD, detection by mass spectrometry (MS) offers better resolution of the components of polysorbate. It also provides increased sensitivity, making it possible to detect subtle changes in concentrations and accurately identify the fatty acids that make up polysorbates (e.g., lauric for PS-20 and oleic for PS-80).
Deployment and scalability
This method is universally applicable for the analysis of polysorbates 20 and 80, regardless of the type of biopharmaceutical product. It is easily deployable in quality control and development laboratories, ensuring flexibility and efficiency for analytical teams.
Versatility
Universal applicability for complex matrices (protein solutions, finished products).
Practical applications in the biopharmaceutical environment
Formulation development: precise monitoring of polysorbate concentrations to optimize protein stability
Stability studies: detection of degradation products to anticipate impacts on drug efficacy
Quality control (QC): monitoring polysorbates in finished products to ensure compliance
In conclusion
Quantitative analysis of polysorbates 20/80 by HPLC-MS is a reference method in the biopharmaceutical sector. Thanks to its sensitivity, specificity, and ability to process complex matrices, it meets the growing demands for drug quality and safety. This technology is an essential tool for analytical and development laboratories in the context of regulatory compliance (EMA, FDA) and technical performance.