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Rapid determination of globin chains in red blood cells by capillary electrophoresis using INSTC oated fused‐silica capillary
Author(s) -
Tůma Petr
Publication year - 2014
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.201400044
Subject(s) - repeatability , capillary action , capillary electrophoresis , chromatography , chemistry , myoglobin , capillary electrochromatography , lysozyme , resolution (logic) , analytical chemistry (journal) , materials science , biochemistry , artificial intelligence , computer science , composite material
A laboratory‐made INSTC oated fused‐silica capillary has been newly used for CE separation of four mixtures of proteins in sodium phosphate BGE s at pH 3.0 and 2.5, respectively. The obtained separation efficiencies range from 145 000 theoretical plates per meter for myoglobin to 1 216 000 m −1 for lysozyme. A total of 49–89% of the number of theoretical plates was obtained in a commercial polyvinyl alcohol coated capillary compared to the INSTC oated capillary under the same experimental conditions, 0–86% was obtained in a laboratory polyacrylamide‐coated capillary, and only 0–6% was obtained in an uncoated fused‐silica capillary. The RSD values for the intraday repeatability for an INSTC oated capillary were 0.1–1.0% (migration time) and 0.3–2.4% (peak area); RSD values for the interday repeatability in the same capillary are 0.6–1.4% (migration time) and 2.4–5.5% (peak area); RSD values for interday repeatability between different capillaries equaled 1.7–2.1% (migration time) and 2.8–10.9% (peak area). The INSTC oated capillary has been further used for rapid determination of globin chains isolated from red blood cells. A separation of α and β chains prepared from adult blood has been completed in 3 min with a peak resolution of 1.3, and the separation of α and G γ chains prepared from newborn blood took 3 min with a peak resolution of 3.6.