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Application of dodecyldimethyl (2‐hydroxy‐3‐sulfopropyl) ammonium in wall modification for capillary electrophoresis separation of proteins
Author(s) -
Wei Wei,
Ju Huangxian
Publication year - 2005
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.200410273
Subject(s) - capillary electrophoresis , chromatography , electrophoresis , chemistry , ammonium , capillary action , materials science , organic chemistry , composite material
A zwitterionic surfactant, dodecyldimethyl (2‐hydroxy‐3‐sulfopropyl) ammonium (C 12 H 25 N + (CH 3 ) 2 CH 2 CHOHCH 2 SO 3 − ), named dodecyl sulfobetaine (DSB), was used as a novel modifier to coat dynamically capillary walls for capillary electrophoresis separation of basic proteins. The DSB coating suppressed the electroosmotic flow (EOF) in the pH range of 3–12. At high DSB concentration, the EOF was suppressed by more than 8.8 times. The DSB coating also prevented successfully the adsorption of cationic proteins on the capillary wall. Anions, such as Cl − , Br − , I − , SO 4 2− , CO 3 2− , and ClO 4 − , could be used as running buffer modifiers to adjust the EOF for better separation of analytes. Using this dynamically coated capillary, a mixture of eight inorganic anions achieved complete separation within 4.2 min with the efficiencies from 24 000 to 1 310 000 plates/m. In the presence of ClO 4 − as EOF adjustor, the separation of a mixture containing four basic proteins (lysozyme, cytochrome c , α‐chymotrypsinogen A, and myoglobin) yielded efficiencies of 204 000–896 000 plates/m and recoveries of 88%–98%. Migration time reproducibility of these proteins was less than 0.5% relative standard deviation (RSD) from run to run and less than 3.1% RSD from day to day, showing promising application of this novel modifier in protein separation.