Basic gel permeation chromatography studies. III. Mathematical analysis of peak spreading

Abstract

Abstract Peak spreading in gel permeation chromatography has been studied with a range of gels including those whose permeation limit corresponded to about 10 3 , 10 6 , 10 8 , and 10 9 molecular weight polystyrene. Peak spreading conformed to the equation Y V 2 = Y M 2 + Y A 2 + Y I 2 + Y D 2 + Y S 2 , where Y V is the peak width of a normal chromatogram, Y M is the contribution due to the true molecular weight of the sample, Y A is due to peak spreading in the apparatus, Y I is spreading in the interstitial volume, Y D is diffusional spreading due to time spent in the gel, and Y S is due to sorption. Evaluating the appropriate parts of the equation leads to measures of the true molecular weight distribution and the contribution due to diffusion into and out of the gel. The data also allowed estimates as to the diffusional spreading with small molecules. With polystyrene having 100 000 molecular weight, diffusional spreading accounts for 80% of Y V , 2 but with small molecules the contribution due to diffusion was not detected.