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High‐resolution dynamic computer simulation analysis of the behavior of sample components with p I values outside the p H gradient established by carrier ampholyte CIEF
Author(s) -
Thormann Wolfgang,
Kilár Ferenc
Publication year - 2013
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.201200499
Subject(s) - chemistry , analytical chemistry (journal) , isotachophoresis , anode , sample (material) , cathode , chromatography , resolution (logic) , constant (computer programming) , electrolyte , electrode , artificial intelligence , computer science , programming language
The behavior of sample components whose p I values are outside the p H gradient established by 101 hypothetical biprotic carrier ampholytes covering a pH 6–8 range was investigated by computer simulation under constant current conditions with concomitant constant electroosmosis toward the cathode. Data obtained with the sample being applied between zones of carrier ampholytes and on the anodic side of the carrier ampholytes were studied and found to evolve into zone structures comprising three regions between anolyte and catholyte. The focusing region with the p H gradient is bracketed by two isotachopheretic zone structures comprising selected sample and carrier components as isotachophoretic zones. The isotachophoretic structures electrophoretically migrate in opposite direction and their lengths increase with time due to the gradual isotachophoretic decay at the p H gradient edges. Due to electroosmosis, however, the overall pattern is being transported toward the cathode. Sample components whose p I values are outside the established p H gradient are demonstrated to form isotachophoretic zones behind the leading cation of the catholyte (components with p I values larger than 8) and the leading anion of the anolyte (components with p I values smaller than 6). Amphoteric compounds with appropriate p I values or nonamphoteric components can act as isotachophoretic spacer compounds between sample compounds or between the leader and the sample with the highest mobility. The simulation data obtained provide for the first time insight into the dynamics of amphoteric sample components that do not focus within the established p H gradient.