STUDY ON HPLC OF AROMATIC AMINO ACIDS AND OLIGO PEPTIDES USING MICROSPHERICAL CARBON PACKINGS : HPLC of Amino Acids and Oligo Peptides on Carbon Column

A microspherical carbon (MSC) as HPLC packings has been tested and its chromartographic propertier for the separation of aromatic amino acids and some oligo peptides have been studied in this paper. Theapparent characterization o packings were described by scanning electron microggraph and physical parameters. The mixture of aramatic amino acids can be separated satisfacterily on MSC column (7.8mm I. D. ×50mm) using acetonitrile eluate of the linear gradient, while the non-aromatic amino acids were not retained under the same conditions. The good results for the separation of some oligo peptides bearing aromatic structure can be obtained, while the other type of oligo peptides were not yet retained on MSC column unless the sample possess rather strong hydrophobicity. The high selective retention is a significant characteristic of MSC packing, and the chromatographic method described is useful for the separation of biochemical substances especially as some oligo peptides from the hydrolysis products of proteins or from the drinks. The separation machanism and exiting problem for MSC packings were also discussed preliminarily

CHROMATOGRAPHY OF PROTEIN ON HYDROXYAPATITE IMPREGNATED PAPER

Ceramic hydroxyapatite beads for high performance liquid chromatography (HPLC) was already reported to be a cation exchanger in type and proteins were adsorped and eluted with changing the concentration of sodium phosphate buffer solution. Hydroxyapatite crystalline impregnated paper (HAP paper) which was made of micro crystalline and paper fiber, but without sintering process was used for filtration or separation of biological macromolecule materials. A circular sheet of HAP paper was cut into a suitable size and (0.76mm I. D.) and placed in a column for HPLC of protein. Chromatography was performed across the HAP paper sheet. One sheet of HAP paper could adsorbed about 12μg cytochrome c. A pile of these sheets could also be used for the chromatography of protein as well as hydroxyapatite beads column. HAP paper was useful not only for the separation of protein but also for the preliminary sample clean up for the HPLC, electrophoresis or various immunochemical treatment