Characterization of reversed-phase high-performance liquid chromatographic stationary phases using ribonuclease A.
The protein ribonuclease A (RNase A) represents a good model protein for studying reversible conformational refolding during gradient elution. Work is described utilizing RNase A under gradient conditions to evaluate several different reversed-phase materials. Columns (10 cm x 4.6 mm I.D.) were packed with Partisil C18, Vydac C18, Nucleosil C4, Nucleosil C18 and an adamantyl-modified Partisil silica. Measurements of the apparent first-order rate constant of refolding, as a function of temperature, are presented and compared for each stationary phase. Comparisons of peak shapes as functions of flow-rate and temperature are also discussed.
Aubel
M T
MT
Department of Chemistry, University of Tennessee, Knoxville 37996-1600.
Guiochon
G
G
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D012259:Ribonuclease, Pancreatic / Q000032:analysis; D012822:Silicon Dioxide; D013696:Temperature
D012822:Silicon Dioxide; D012259:Ribonuclease, Pancreatic