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Based on chromatographic theory, resolution and column length is related by a square root function.This square root relationship means that a large length of column has to be removed before a significant loss in resolution occurs. For a 30-m column, a loss of 1 m results in only a 1.7% decrease in resolution.

Without having to redevelop the method, the increased resolution can readily be achieved by increasing the column length. A disadvantage of using longer columns is that the analysis time has to be increased accordingly.

As the resolution is proportional to the square root of the number of theoretical plates in equation (1), doubling the number of theoretical plates increases the resolution by a factor of 221a2ffe3= 1.41; increasing the number of theoretical plates four times increases the resolution by a factor of 221a4ffe3= 2.

Resolution. The resolution of a elution is a quantitative measure of how well two elution peaks can be differentiated in a chromatographic separation. It is defined as the difference in retention times between the two peaks, divided by the combined widths of the elution peaks.

Equation (1) indicates that the resolution is the difference between peak retention times divided by the average peak width. In a peak with Gaussian distribution, the peak width is W = 4 03c3 (where 03c3 is the standard deviation) and the peak FWHM is W0.

Resolution is an important HPLC performance indicator usually assessed by how quickly and how completely target components in a sample separate as they pass through a column. Resolution is measured by dividing the difference in peak retention times by the average peak width.

How to Improve Resolution in HPLCIncreasing column length.Decreasing particle size.Reducing peak tailing.Increasing temperature.Reducing system extra-column volume.