Preparation of an ion-exchangeable polymer bead wrapped with bilayer membrane structures for high performance liquid chromatography.
ABSTRACT
We synthesized a chromatographic packing material that has a non-covalently
attached dihexadecyl phosphate (DHP) bilayer membrane structure on a CA08S,
a nonporous-type cationic polymer bead with a diameter ranging from 11
to 14 ƒÊm. Confocal fluorescence microscopic and differential scanning calorimetric
analyses of the DHP-CA08S complex revealed that the DHP bilayer membrane
structures were formed on the surface of the CA08S polymer beads. When
the functionality of the DHP-CA08S complex was evaluated in the ion-exchange
HPLC of proteins, the retention behavior of the proteins on the DHP-CA08S
complex column totally mirrored the anionic property of the DHP bilayer
membrane surface, not the cationic property of the CA08S bead. Methylene
blue (MB) was eluted from the DHP-CA08S complex column in the isocratic
elution mode, but not at all from a CK08S column, a styrene-divinylbenzene
based cation-exchange polymer. When the column temperature was elevated
from 50 to 60 Ž, the peak shape of MB on the DHP-CA08S complex column became
fairly sharp without a change in its peak area, which mirrored the characteristic
phase transition of the DHP bilayer membrane formed on the DHP-CA08S complex.