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Bulletin of the Korean Chemical Society (BKCS)

ISSN 0253-2964(Print)
ISSN 1229-5949(Online)
Volume 14, Number 4
BKCSDE 14(4)
April 20, 1993 

 
Title
A Study of the Retention Behavior of Proteins in High-Performance Liquid Chromatography(Ⅰ) : The Effect of Solvent and Temperature on Retention Behavior of Proteins in Reversed-Phase Chromatography
Author
Dai Woon Lee*, Byung Yun Cho
Keywords
Abstract
The retention behavior of proteins was investigated by using reversed-phase chromatography (RPC), comparing to the retention behavior of small molecules in RPC. The evaluation was carried out on a SynChropak RP-P(C18) column with 0.1% aq. TFA-organic solvent modifier such as acetonitrile, isopropanol, and ethanol. The Z value (the number of solvent molecules required to displace the solute from the surface) was a general index for the characterization of protein retention as a function of organic concentration over a range of temperature between 5 and 70℃. Van't Hoff plots provided the basis for evaluating the enthalpic and entropic changes associated with the interaction between protein and the stationary phase. Z values did not change significantly at the range of temperature showing the consistent △H° and △S° values. From these investigation, it was concluded that the retention behavior of proteins in RPC was able to be predicted by the retention parameters applied to small molecules. Furthermore, myoglobin and hemoglobin in RPC as stated above showed a similar retention behavior regardless of their molecular weights.
Page
510 - 514
Full Text
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