Novel cationic coating agent for protein separation by capillary electrophoresis(†).
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.
Regional Centre of Advanced Technologies and Materials Department of Analytical Chemistry, Faculty of Science, Palacký University in Olomouc, Olomouc, Czech Republic.
Joanna
J
Znaleziona
Department of Organic Chemistry, Faculty of Science, Charles University in Prague, Prague 2, Czech Republic.
Dušan
D
Drahoňovský
Department of Inorganic Chemistry, Faculty of Science, Palacký University in Olomouc, Olomouc, Czech Republic.
Bohuslav
B
Drahoš
Regional Centre of Advanced Technologies and Materials Department of Analytical Chemistry, Faculty of Science, Palacký University in Olomouc, Olomouc, Czech Republic.
Juraj
J
Ševčík
Regional Centre of Advanced Technologies and Materials Department of Analytical Chemistry, Faculty of Science, Palacký University in Olomouc, Olomouc, Czech Republic.
Vítězslav
V
Maier
D002412:Cations / Q000737:chemistry; D045304:Cytochromes c / Q000737:chemistry / Q000302:isolation & purification*; D019075:Electrophoresis, Capillary; D009113:Muramidase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D009211:Myoglobin / Q000737:chemistry / Q000302:isolation & purification*; D012259:Ribonuclease, Pancreatic / Q000737:chemistry / Q000302:isolation & purification*; D012822:Silicon Dioxide / Q000737:chemistry*; D013501:Surface-Active Agents / Q000737:chemistry*
D002412:Cations; D009211:Myoglobin; D013501:Surface-Active Agents; D012822:Silicon Dioxide; D045304:Cytochromes c; D012259:Ribonuclease, Pancreatic; D009113:Muramidase
Capillary electrophoresis; Cationic surfactants; Dynamic coating; Lysozyme; Proteins