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Novel cationic coating agent for protein separation by capillary electrophoresis †
Author(s) -
Znaleziona Joanna,
Drahoňovský Dušan,
Drahoš Bohuslav,
Ševčík Juraj,
Maier Vítězslav
Publication year - 2016
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201501349
Subject(s) - lysozyme , chemistry , chromatography , pulmonary surfactant , capillary electrophoresis , bromide , ammonium bromide , myoglobin , cationic polymerization , chloride , ammonium chloride , analytical chemistry (journal) , inorganic chemistry , organic chemistry , biochemistry
A novel positively charged surfactant N ‐dodecyl‐ N , N ‐dimethyl‐(1,2‐propandiol) ammonium chloride was used for the dynamic coating of the inner wall of a silica capillary. This paper covers the evaluation of dynamic coating and study of the influence of the analysis conditions for the magnitude and direction of electroosmotic flow as well as for the effective and selective separation of chosen proteins (ribonuclease A, cytochrome c, lysozyme, and myoglobin). The concentration of 0.1 mM of N ‐dodecyl‐ N , N ‐dimethyl‐(1,2‐propandiol) ammonium chloride enabled the reversal of the electro‐osmotic flow, however, to separate basic as well as neutral proteins the higher concentration of the studied surfactant was necessary. The final conditions for the separation of studied proteins were set at 100 mM sodium acetate pH 5.5 with 10.0 mM of the studied surfactant. The results were also compared with those of two commercially available cationic surfactants, cetyltrimethylammonium bromide and dodecyltrimethylammonium bromide. Additionally, the developed method for protein separation was applied for the determination of lysozyme in a cheese sample. The limits of detection and quantification of lysozyme were 0.9 and 3.0 mg/L, respectively. The mean concentration of lysozyme found in the cheese sample was 167.3 ± 10.3 mg/kg.