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What is resolution in ion exchange chromatography?

Resolution in ion exchange chromatography The result of an ion exchange experiment, as with any other chromatographic separation, is often expressed as the resolution between the peaks of interest. The resolution is defined as the distance between peak maxima compared with average base width of the two peaks.

How does Column length affect resolution in GC?

It is a well-known fact that the separation efficiency of a chromatographic column increases with an increase in its length. Doubling the column length increases resolution by around 40%.

How does column length affect resolution?

Chromatographic parameters The advantage of switching to longer columns is that the theoretical plates will increase and so will the resolution. The number of theoretical plates (N) is directly proportional to the length of the column and inversely proportional to the particle diameter2.

What is column length in HPLC resolution?

HPLC Instrument Considerations Standard columns for reversed phase, normal phase, and ion exchange chromatography typically range from 3.9 to 4.6 mm internal diameter and 15, 25, and 30 cm in length. Standard columns for gel permeation chromatography (GPC) are 7.8 mm by 30 cm.

What is the effect of using a longer column?

Well, longer columns do provide higher plate counts (more plates per column) meaning that the elution bands are sharper (reduced fraction volume) which is what you want when performing analytical HPLC. Wider columns, however, provide increased loading capacity and this is what preparative chromatography needs.

What is the polishing step in a protein purification strategy?

Polishing: The main aim of this polishing step is to remove minor contaminants and also aggregates present in the mixture. This step is also used to transfer the purified sample into suitable buffer, which can be used for further structural studies.

What does increasing column length affect chromatography?

A longer column generally improves the separation. The trade-off is that the retention time increases proportionally to the column length and a docHub peak broadening will be observed as well because of increased longitudinal diffusion inside the column.

How can the resolution of ion chromatography be improved?

The easiest way to improve the resolution from an IEX run is to modify the running conditions. Different pH will affect the charged surface on the protein, which affects the resolution. A smaller amount of sample will improve the resolution: typically use up to 30% of the complete capacity to maintain good resolution.

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Polish columns resolution easily