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Bulletin of the Korean Chemical Society (BKCS)

ISSN 0253-2964(Print)
ISSN 1229-5949(Online)
Volume 27, Number 9
BKCSDE 27(9)
September 20, 2006 

Elution Behavior of Protein and Pullulan in Asymmetrical Flow Field-flow Fractionation (AsFlFFF)
Eunsun Ji, Seong-Ho Choi, Kuk Ro Yoon, Jong-Han Chun, Seungho Lee*
Asymmetrical flow-field flow fractionation (AsFlFFF), Protein, Pullulan, Plate height, Calibration
An AsFlFFF channel was designed and built, and then tested for analysis of pullulans and proteins. Pullulans and proteins having various nominal molecular weights were injected at various conditions of the cross-flow rate (Fc) and the channel-out flow rate (Fout). The retention (measured by the retention ratio R) and the zone broadening (measured by the plate height H) were measured, and then compared with theory. When the incoming flow rate, Fin (and thus Fout) was varied with Fc fixed at 2.5 mL/min, the plate height measured for the pullulan with nominal molecular weight (M) of about 100,000 showed the trend expected by the “longitudinal diffusion” theory (H decreases with increasing flow rate). In contrast, when Fout was varied with the flow rate ratio, Fc/Fout, fixed constant at 5, the plate height measured for the same sample showed the trend expected from the “non-equilibrium” theory (H increases with increasing flow rate). Calibration plots (log D vs. log M) obtained with pullulans and proteins were not coincide, probably due to the difference in molecular conformation, suggesting the analysis of pullulans and proteins using AsFlFFF requires independent calibration. It was found that the linearity of the protein-calibration plot was improved by using a buffer solution as the carrier.
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