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Correlation of capillary zone electrophoresis with continuous free‐flow zone electrophoresis: Application to the analysis and purification of synthetic growth hormone releasing peptide
Author(s) -
Prusik Zdeněk,
Kašička Václav,
Mudra Petr,
Štěpánek Jiří,
Smékal Otto,
Hlaváček Jan
Publication year - 1990
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150111109
Subject(s) - capillary electrophoresis , chromatography , chemistry , electrophoresis , free flow electrophoresis , peptide , fractionation , analytical chemistry (journal) , acetic acid , capillary action , gel electrophoresis of proteins , polyacrylamide gel electrophoresis , biochemistry , materials science , composite material , enzyme
Abstract Two carrier‐free electrophoretic separation methods, capillary zone electrophoresis (CZE) and continuous free‐flow zone electrophoresis (FFZE), have been applied to both microanalysis at the nanogram level and preparative fractionation, with a throughput of 30 mg/h, of synthetic growth hormone releasing peptide (GHRP). A crude product of GHRP, a hexapeptide with the sequence His‐D‐Trp‐Ala‐Trp‐D‐Phe‐Lys‐NH 2 , synthesized by the solid phase methodology, was desalted and analyzed by CZE. Based on the results of analytical CZE the separation was converted into a preparative purification procedure by continuous FFZE, employing the same separation medium (0.5 mol/L acetic acid, pH 2.6). The purity of peptide frcttions obtained by FFZE was reevaluated by CZE. The combination of these two techniques proved to be a valuable tool for both peptide analysis and peptide purification. A close correalation of CZE and FFZE, resulting from the fact that both methods are based on the same separation principle (zone electrophoresis) and that both are performed in a free solution of the same composition, was confirmed. However, when transforming data from CZE to FFZE, the different electroosmotic flow, temperature and electric field intensity in the capillary and in the flow‐through cell, respectively, have to be taken into account and corresponding corrections have to be made.