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Dual stacking of unbuffered saline samples, transient isotachophoresis plus induced pH junction focusing
Author(s) -
Shim SangHee,
Riaz Asif,
Choi Kihwan,
Chung Doo Soo
Publication year - 2003
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.200305411
Subject(s) - isotachophoresis , stacking , electrolyte , chemistry , analytical chemistry (journal) , chromatography , dilution , capillary electrophoresis , chloride , analyte , transient (computer programming) , ion , electrode , thermodynamics , physics , organic chemistry , computer science , operating system
A dual stacking mechanism based on transient isotachophoresis (TITP) and induced pH junction focusing is demonstrated as a means to increase the concentration sensitivity in capillary electrophoresis of highly saline samples. When stacking was carried out with an unbuffered saline sample of fluorescein between two zones of low mobility background electrolyte at high pH under an electric field of reverse polarity, two transient peaks at both boundaries of the sample zone were observed. One peak at the rear boundary could be inferred as a transient isotachophoretic stacked zone. Through computer simulations of an unbuffered sample with a high concentration of sodium chloride, we showed that the fast moving zones of sodium and chloride ions induced pH changes at both boundaries to satisfy the electroneutrality condition and that the peak at the front boundary was due to the induced pH junction. To verify the pH changes, an indicator, thymol blue, was added to an NaCl solution and the color changes under an electric field were observed. The proposed mechanism was supported by observing the dual stacking procedure for an unbuffered sample of 4‐nitrophenol and measuring additional sensitivity enhancements by dual stacking for ten weakly acidic compounds. For the ten analytes including nucleoside phosphates, every dual stacking of an unbuffered sample exhibited an additional enhancement up to 86% larger than that of usual transient isotachophoresis of the corresponding buffered sample without loss of separation efficiency and reproducibility. Therefore, it would be useful to skip over buffering in sample preparation for TITP, contrary to the general recommendation.