Gel permeation chromatography - Mass Spectrometry (GPC-MS)

Gel permeation chromatography (GPC), also known as size-exclusion chromatography (SEC), is an analytical technique used to characterize various polymers or other macromolecules in a mixture. However, conventional GPC using a single concentration-sensitive detector may have limitations and drawbacks. GPC coupled mass spectrometry (MS) is a powerful analytical method that combines the separation abilities of gel permeation chromatography with the detection and identification capabilities of mass spectrometry, allowing the analysis of complex mixtures with high resolution and sensitivity. GPC-MS technology can be widely used to analyze complex macromolecule mixtures, such as proteins, peptides, oligonucleotides, and polymers, based on their molecular size and structure.

Principle of GPC-MS

• GPC is a separation based on molecular size rather than chemical properties. It uses the principle of size exclusion chromatography to separate samples of polydisperse polymers into components with a narrow molecular weight distribution. This separation is achieved by using a porous stationary phase, which allows smaller molecules to enter the pores and take longer to elute.
• In mass spectrometry, eluted molecules are ionized, and mass analyzed, allowing identification based on their mass-to-charge ratio (m/z).

Applications of GPC-MS

GPC-MS is capable of analyzing complex mixtures with high resolution and sensitivity, making it a valuable tool in various fields such as protein analysis, polymer characterization, oligonucleotides, biomedical research, and more.

• Protein Analysis: GPC-MS can be used for the analysis of proteins and peptides to determine their molecular weight, oligomer status and post-translational modifications, which are crucial for studying protein function and stability.
• Polymer Characterization: GPC-MS separates polymers based on size, allowing researchers to analyze their molecular weight distribution, branching, and end-group functionality. These are crucial for quality control in polymer production and for understanding the structure-property relationships of polymers.
• Oligonucleotide Analysis: GPC-MS can be used to analyze oligonucleotides such as DNA and RNA. By separating these biomolecules according to size, researchers can elucidate their sequences, modifications, and interactions with proteins or small molecules.
• Biomedical Research: GPC-MS can be used in various areas of biomedical research, such as metabolomics, proteomics and biomarker discovery. By GPC-MS, researchers can identify and quantify biomolecules in complex biological samples.

Notes

While there are challenges in coupling GPC with MS. The biggest one is the incompatibility of mobile phases containing high concentrations of non-volatile salts. These mobile phases lead to ion suppression and contamination of the mass spectrometer. To overcome this difficulty, GPC-MS methods, which use denaturing mobile phases containing organic solvents and ion-pairing reagents, have been developed^[1].

Alfa Chemistry is a leading company in the field of analytical chemistry with a team of experienced professionals who are experts in various analytical techniques. Our GPC-MS platforms ensure consistently accurate and detailed delivery. Alfa Chemistry looks forward to cooperating with you.

Reference

1. Hong P, Koza S, Bouvier ES. Size-Exclusion Chromatography for the Analysis of Protein Biotherapeutics and their Aggregates. J Liq Chromatogr Relat Technol. 2012 Nov;35(20):2923-2950.

Our services are for research use only.