Abstract
The various factors that affect the loading capacity of a column are considered and it is shown that large-scale separations can only be achieved at the individual or combined expense of speed, scope and solvent consumption. An equation that predicts the maximum sample volume that can be placed on a column without impairing resolution is derived and experimentally verified. It is shown that, when the sample volume is progressively increased, the fronts of all peaks remain at constant retention while the retention of the rears of the peaks progressively increases. Conversely, under circumstances of mass overload the rears and fronts of all peaks tend to move toward positions of less retention, the maximum effect occurring with the major component in the mixture.
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