5 Factors to Be Considerd When Selecting Flash Column
5 Factors to Be Considered When Selecting Flash Column
Selection of Flash Column
Reliable flash columns play a decisive role in a good chromatographic separation method, which ensures the complete separation of valuable compounds and specific impurities. Choosing the right flash column is the most critical part of the purification process. There are two aspects that should be carefully considered to select an appropriate flash column.
Stationary Phase of Flash column
To determine the stationary phase of the flash column, there are three aspects that should be considered, including matrix type, bonded phase, and matrix parameters. The commonly used matrix materials are divided into three categories – inorganic materials, organic materials, and composite materials. Silica, with high mechanical strength and thermal stability, is the most widely used inorganic matrix in the flash column. It holds an important position in the separation and purification of compounds in the normal phase purification system. Nevertheless, its stability is relatively poor under alkaline conditions.
The organic matrix is widely applied in the separation and purification of biomolecules such as proteins and sugars. As for the bonded phase, different separation types of stationary phases could be formed by bonding different groups to the matrix, such as stationary phase, reverse phase, ion exchange, SEC, and the like. The matrix parameters, including the shape, particle size, and pore size of the matrix, are selected in terms of the specific needs of the purification. Such as Superior Silica Gel Flash Column, Spherical HILIC Flash Column, and Neutral Alumina Flash Column.
Compatibility of Flash column
Generally, the flash column is made of polypropylene material in a full range of size selections, which could satisfy most customers’ demands. Considering the Luer-Lok outlet fittings are available to support any flash column chromatography systems, it would be your first choice.
HAWACH has summarized 5 important factors to be considered when you gonna choose the Flash column.
1. Packing material matrix 1.1 Silica gel matrix: high purity, low cost, high strength, easy chemical modification, but with limited pH range. Most silica-based packing materials are stable between pH 2-8, but the specially modified silica-bonded phase can be stabilized at pH 1.5-10. 1.2 Polymer matrix: wide application pH range, stable temperature (high temperature can reach above 80 degrees), low mechanical strength.
2. Particle 2.1 Particle shape Spherical particles provide lower column pressure, higher efficiency and stability, and longer column life when using a high-viscosity mobile phase; irregular particles have a larger specific surface area and a relatively low price.
2.2 Particle size The smaller the particle size, the higher the column efficiency, and the higher the resolution, but at the same time, it will result in a higher column pressure drop. Choose 1.5-3μm packing to solve some complex samples. UPLC can use 1.5μm packing; another 10μm or larger packing can be used as a semi-preparative or preparative column.
3. Carbon content The carbon content refers to the proportion of the bonded phase on the surface of silica gel, which is related to the specific surface area and bonding coverage. High carbon content improves column capacity, resolution, and analysis time, which is suitable for complex samples requiring high resolution; low carbon content’s analysis time is short, exhibits different selectivity, and is used for rapid analysis of simple samples and samples with high aqueous active phase conditions. Generally, the carbon content of C18 varies from 7-19%.
4. Pore size and specific surface area Pore diameter and specific surface area are two concepts that complement each other. Small pore size and large surface area, and vice versa. Large specific surface area increases the reaction between the sample and the bonded phase, and increases storage, sample loading, and resolution of complex components; the small specific surface area has a fast equilibrium time, suitable for gradient analysis.
5. Pore volume and mechanical strength Pore volume, also known as “pore volume” refers to the size of the void volume per particle. It can well reflect the mechanical strength of the packing. Packing with a large pore volume has a slightly weaker mechanical strength than packing with a smaller pore volume. Packings with a pore volume greater than 1.5 mL/g are used for size-exclusion chromatography and low-pressure chromatography.