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Trends in Chromatography   Volumes    Volume 1 
Abstract
A novel zonal electrophoresis in a porous glass sheet made by high technology
Masanori Yoshioka, Tadashige Chiba, Toshihide Takagi, Hiroshi Kita, Misao Seto
Pages: 83 - 87
Number of pages: 5
Trends in Chromatography
Volume 1 

Copyright © 2005 Research Trends. All rights reserved

ABSTRACT

Previously based on high technology, we developed porous glass sheets with pore diameters from 110 to 1200 nm for use in thin layer chromatography. A   sheet of 730 nm pore diameter among the various diameters showed rapid and best separation of inorganic and organic compounds in a few minutes.  In this paper, the sheets were used as a supporting medium for electrophoresis. The pores were completely uniform in size to make long winding capillaries for any directions just like a limestone cave. Thus, a zonal electrophoresis was carried out in the sheet, in which spots were detected by means of conventional zonal electrophoresis. Separation condition for standard amino acids was examined in buffers for isocratic or isoeletric focusing electrophoresis. The sheet of 730 nm was also best for the electrophoresis. On a rectangle of 2.5 cm X 1.0 cm of glass sheet of 0.5 mm thick, basic lysine, alanine and acidic glutamic acid were separated in 10 mM citrate buffer at pH 5.0 at 9 V for 20 minutes. The minimum detectable amounts of amino acids were 200 pmol by color reaction with ninhydrin and 10 pmol by fluorescence reaction with fluorescamine. Three amino acids were also separated in 5 mM phosphate buffer at 15 V for 30 minutes in a simple buffer without any polyampholyte by isoelectric focusing. The most convenient merit of the sheet was the minimum electric resistance of the buffer, which enabled low-voltage electrophoresis by a small and portable apparatus with dry batteries at a few voltages. It is to say that the present zonal electrophoresis is a kind of capillary or frontal electrophoresis in principle.

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