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Why is sample volume a critical factor in planning a gel chromatography experiment? What would be...

Why is sample volume a critical factor in planning a gel chromatography experiment? What would be the percentage of sample volume in column total volume?

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Answer #1

The sample volume is a critical factor in planning a gel chromatography experiment.If too much sample is applied to a column, resolution is decreased; if the sample size is too small, the solutes are greatly diluted.For group separations, a sample volume of 10 to 25% of the column total volume is suitable.The sample volume for fractionation procedures should be between 1 and 5% of the total volume.Column total volume is determined by measuring the volume of water in the glass column that is equivalent to the height of the packed bed.

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