GFC/SEC is a non-interactive chromatographic technique that separates analytes solely according to their molecular size and shape. The column packing consists of porous particles that provide a well-defined pore structure. Large molecules that cannot enter the pores elute first, while smaller molecules penetrate deeper into the pore network and elute later.
Because there are no adsorption, ion-exchange, or hydrophobic interactions, GFC/SEC preserves the native structure of the analytes and provides highly reproducible separations. This makes it the preferred method for determining molecular weight distributions, detecting aggregates, and analyzing molecular conformations in aqueous systems.
Depending on the column material, GFC/SEC can be used for both biomolecules and synthetic water-soluble polymers, offering excellent stability and long-term performance even under routine analytical or preparative conditions.
Gel Filtration Chromatography is widely employed for the separation and analysis of proteins, polysaccharides, and hydrophilic polymers. The technique is mild, highly reproducible, and independent of analyte charge or polarity.
It provides valuable information on molecular size, aggregation behavior, and sample homogeneity, and is a standard method for purification and molecular weight determination in biochemistry and polymer analysis.
The stationary phase in GFC/SEC consists of porous silica gels and crosslinked methacrylate polymer particles, both engineered for precise pore size control and excellent mechanical stability.
Silica-based materials offer high efficiency, narrow particle size distribution, and excellent column rigidity, making them suitable for high-performance aqueous SEC applications. Methacrylate polymer gels, on the other hand, provide superior chemical stability and hydrophilicity, minimizing non-specific interactions and ensuring long column lifetime.
Both materials enable pure size-based separation without secondary effects, allowing accurate molecular size characterization across a broad range of molecular weights.
In GFC/SEC, the mobile phase functions purely as a solvent medium and does not participate in chemical interactions with the analytes. Typically, deionized water — optionally containing small amounts of organic modifiers such as methanol or ethanol — is used.
Since no buffering components are required, the method avoids ionic effects and ensures straightforward, highly reproducible separations. The choice of solvent is governed by analyte solubility and compatibility with the stationary phase, ensuring stable baseline conditions and excellent detector response.
The retention mechanism in Gel Filtration Chromatography is based entirely on size exclusion. Molecules larger than the pore diameter are excluded and elute first, while smaller molecules enter the pores and elute later depending on their hydrodynamic volume.
This purely physical mechanism guarantees non-destructive, reproducible separations that reflect true molecular dimensions. Because no chemical interactions occur, GFC/SEC provides reliable molecular size characterization for a wide variety of water-soluble samples.
| Name | Particle Sizes | Carbon Load | Pore Size |
|---|---|---|---|
| GromSil 120 SEC | 3 – 5 µm | 120 Å | |
| GromSil 200 SEC | 5 µm | 200 Å | |
| GromSil 300 SEC | 5 µm | 300 Å | |
| Macrosphere 60 SEC | 5 µm | 60 Å | |
| Macrosphere 60 Si | 5 µm | 0 % | 60 Å |
| Macrosphere 100 SEC | 5 µm | 100 Å | |
| Macrosphere 100 Si | 3 – 5 µm | 0 % | 100 Å |
| Macrosphere 200 SEC | 1.9 – 5 µm | 200 Å | |
| Macrosphere 300 SEC | 5 µm | 300 Å | |
| Macrosphere 400 SEC | 5 µm | 400 Å | |
| Macrosphere 800 SEC | 5 µm | 800 Å | |
| ReproGel 100 RP-1 | 5 – 10 µm | ||
| Repromer OH-2500 | 10 µm | ||
| Repromer OH-3000 | 10 µm | ||
| Repromer OH-4000 | 10 µm | ||
| Repromer OH-5000 | 10 µm | ||
| ReproSil 50 SEC | 5 – 10 µm | 50 Å | |
| ReproSil 125 SEC | 3 – 10 µm | 125 Å | |
| ReproSil 200 SEC | 1.9 – 10 µm | 15.5 % | 200 Å |
| ReproSil 200 SEC-2 | 1.5 – 10 µm | 200 Å | |
| ReproSil 300 SEC | 3 – 5 µm | 300 Å | |
| ReproSil 4000 SEC | 5 µm | 11 % | 400 Å |
| ReproSil 5000 SEC | 5 µm | 5 % | 800 Å |