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Assessing the Degradation of Biopharmaceutical Compounds by Liquid Chromatography Coupled Mass Spectrometry (LC-MS) (CAT#: STEM-B-0404-CJ)

Introduction

All biopharmaceutical compounds are sensitive towards chemical degradation. Examples are deamidation, hydrolysis, oxidation, photo-degradation, disulfide-scrambling, and others. Chemical degradation can lead to aggregation, charge variants and/or structural changes of the drug substance and eventually impair the effectivity or safety of the therapy.




Principle

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.

Applications

Biopharmaceutica

Procedure

1. Sample preparation: Total LC/MS sample preparation must take into consideration both the mobile phase (containing the analyte) and stationary phase (packing material or support). Soluble and insoluble matrix components should also be considered and whether they might interfere with the analyte's final elution from the column.
2. Samples prepared for injection into liquid chromatography MS.
3. Lectrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI) is typically applied to generate charged ions from the sample, and it is these ions that pass into the mass analyzer.
4. The identity of the target ion is determined by comparing its mass:charge (m/z) ratio against a spectral database such as MassBank, METLIN and mzCloud.

Materials

• Sample: Peptides, Proteins, Vaccines, Virus-like particles, Antibody
• Equipment: Liquid chromatography, Mass spectrometry devices, Interface
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