what is hplc

Effect of flow rate on column chromatography

Column chromatography is a widely used technique for separation and purification of compounds from the complex mixtures. In which the compounds pass through the stationary phase with the help of mobile phase and get separate on the basis on varying degrees of adhesion.There is an optimal solvent flow rate for each column, it relies on which type of mobile phase, analyte, and column dimension are being used. If the solvent flow rate is too fast, is not getting adequate time to equilibrium for the compounds and will be forced down the column with leaving a long tail. If the solvent flow rate is used very slowly in column chromatography, the diffusion processes will lead to band widening.

For columns of smaller diameter, the optimal rate is lower than columns with a larger diameter. Therefore, compared with smaller columns the larger columns can be run with the higher flow rate.We know that the size of the collected fractions relies on Rf value and size of the column and that the flow rate of the solvent can affect the separation process.

The suitable flow rate in column chromatography is dependent on the dimensions of the column. The major effect of flow rate on column chromatography is that changing the flow rate of the analysis can change the separation quality of the component.


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