Parameters of HPLC Columns
In HPLC columns, silica gel is the most universal matrix, the strength is large, the chemical modification is easy, but the pH value is limited; the chemical properties of the polymer matrix are stable, and the pH range is wide, but the strength is not large enough, the batch repeatability is relatively poor, and the price is slightly higher. Additionally, the smaller the particle size, the faster the separation, the higher the column efficiency.
In HPLC, the chromatography packing you choose has a great influence on the separation effect. However, the range of chromatographic packing choices is huge. Consequently, if you want to make your best choice, you must have some understanding of chromatographic packing.
Silica gel for HPLC packing
Generally speaking, the stationary phase of normal-phase chromatography is silica and some other amine. Because the SiOH on the surface of silica and other polar groups have great polarity, so we separate according to the polarity of each component, which means the one with the weakest polarity would be flushed out of the column. Comparing to the stationary phase, the mobile phase has weaker polarity.
Inversed phase chromatography uses silica as a matrix, with weaker polarity surface bond. The mobile phase in Inversed Phase Chromatography has great polarity. However, the components with great polarity would be flushed out, and other components with small polarity would be reserved.
Polymer for HPLC packing
Usually, polymers for HPLC packing are polymethacrylate. One of its great advantages is that all PH ranges can use. Comparing to silica gel packing, the polymer has stronger hydrophobic properties and works well on separating the protein samples. However, the shortage is the column efficiency is too low.
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