Analysis of Surfactants Quantification and Characterisation by Mass Spectrometry (MS) Combined with Liquid Chromatography (LC) (CAT#: STEM-B-0369-CJ)

Mass spectrometry separates ionized molecules according to their mass-to-charge ratio. In combination with a-priori knowledge on the sample, LC-MS analysis allows to obtain a comprehensive insight of your sample on a molecular level. With high-resolution quadrupole time-of-light (TOF) mass analyzers, coupled with a Ultra-High-Performance Liquid Chromatography (UPLC), various analytical questions can be targeted. Most important for pharmaceutical industries are the characterization and quantification of active pharmaceutical ingredients (API) (of small and large molecule scale) as well as formulation excipients such as surfactants (polysorbates (PS)), saccharides and amino acids.

Surfactants are in many cases an integral part of biopharmaceutical formulations and crucial for API (e.g. protein) stability during pharmaceutical product shelf life. One of the most common surfactants used in biologics is polysorbate 20 (PS20) or polysorbate 80 (PS80) both are typically used to prevent protein absorption on surfaces and protein aggregation induced by interfacial stresses. Yet, PS degradation can also lead to formation of particles. Major root causes for PS degradation are i) the inherent tendency for auto-oxidation and ii) residual host cell membrane proteins with enzymatic activity that hydrolyze the PS ester bonds. Therefore, close monitoring of PS throughout the shelf life of the pharmaceutical product is essential. LC-MS can also be applied to monitor small molecule APIs and to characterize degradation products “popping up” during stability studies with help of tandem MS.

Biopharmaceutica

1. Sample preparation
2. LC causes physical separation of analytes in solution of liquid samples or solid samples.
3. LC detection using MS.
4. LC-MS data plotting.

• Sample: Surfactants (such as: integral part of biopharmaceutical formulations), polysorbate 20 (PS20), polysorbate 80 (PS81), Buffer excipients (such as: amino acids or saccharides), Small molecule APIs
• Equipment: Tandem quadrupole

• The quality requirements for pharmaceutical grade polysorbate 20 and 80 are specified in the different pharmacopoeias. For polysorbate 80, the Chinese Pharmacopoeia describes the strictest regulations for the fatty acid composition and requests an all-oleate polysorbate (≥ 98%), whereas the European and US pharmacopoeias specify a purity of ≥ 58% for oleic acid.
• LC-MS is suitable for the analysis of polar and non-polar compounds, as well as thermolabile molecules. These compounds can range from low molecular mass analytes with m/z values < 1000 Da, to very high molecular mass proteins with m/z values > 100,000 Da.
• LC-MS instruments are expensive to own, operate and maintain. Expertise is required to run the instruments and analyze the data. The sample throughput is moderate in comparison to other analytical techniques.
• As a mass spectrometer is a destructive detector, care has to be taken when handling samples that may not be readily available or that are not obtainable in large amounts. As a laboratory-based rather than in-field technique, the analysis of unstable or reactive samples by LC-MS can prove challenging.