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Elution mechanism of polypeptides in reversed‐phase liquid chromatography based on the critical threshold of organic solvent to induce abrupt change in adsorption capacity to the column packing
Author(s) -
Goda Ryoya,
Sudo Kenichi
Publication year - 2008
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.899
Subject(s) - chemistry , elution , column chromatography , solvent , chromatography , adsorption , phase (matter) , high performance liquid chromatography , organic solvent , reversed phase chromatography , thermoresponsive polymers in chromatography , chromatography column , organic chemistry , chemical engineering , engineering
Adsorption capacity of polypeptides to the column packing in a solution containing multiple organic solvents was found to be expressed by means of an f n value, which is the sum of the ratios of the content of each organic solvent in the solution to the critical content of each organic solvent to cause abrupt change in the adsorption capacity, and to change abruptly at the point where the f n value becomes 1. Additionally, our results indicate that each polypeptide is eluted by the eluent containing a specific organic solvent content regardless of gradient elution rate in reversed‐phase liquid chromatography, and that total organic solvent content in the eluent containing polypeptides is equal to the critical content. Considering the power law relationship between the retention times and the gradient elution rates, our results suggest that the elution of each polypeptide in reversed‐phase liquid chromatography is mainly controlled by abrupt change in the adsorption capacity induced by change in the organic solvent content of the eluent during a gradient elution process, and that the abrupt change repeats across the critical threshold while a polypeptide moves through the column, and as a result, each polypeptide is concentrated in the eluent with the critical threshold. Copyright © 2007 John Wiley & Sons, Ltd.