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Glycobiology Advance Access published online on June 22, 2005
Glycobiology, doi:10.1093/glycob/cwi096
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© The Author 2005. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oupjournals.org
Received May 12, 2005
Revised June 1, 2005
Accepted June 14, 2005

Article

Chromatographic resolution of glucosidic compounds, ginsenosides on polyethersulphone membrane and its application to the quantitative immunoassay for ginseng saponins

Osamu Morinaga 1, Noriko Fukuda 2, Hiroyuki Tanaka 2, and Yukihiro Shoyama 2*

1 Innovation Plaza Fukuoka, Japan Science and Technology Agency, 3-8-34 Momochihama, Sawara-ku, Fukuoka 814-0001, Japan
2 Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan

* To whom correspondence should be addressed.
Yukihiro Shoyama, E-mail: shoyama{at}phar.kyushu-u.ac.jp


  Abstract

A method has been devised for the chromatographic resolution of glucosidic compounds, ginseng saponins on polyethersulphone (PES) membrane. The method results in good resolution and quantitative immunoassay for ginsenoside Rb1 (G-Rb1), -Rc and -Rd in crude extracts of various ginsengs. Newly established method is simpler and applies for quantitative analysis. Ginsenosides developed by acetonitrile/water/acetic acid solvent system on a PES membrane were directly treated with a NaIO4 solution followed by bovine serum albumin (BSA), resulting in a ginsenoside-BSA conjugate on a PES membrane. Anti-G-Rb1 monoclonal antibody (MAb) was bound and, then a second antibody labeled with peroxidase directed against first antibody. Finally a substrate reacted to the enzyme and gave staining. The stained membrane was scanned and coloring spots were analyzed quantitatively using graphic analysis of NIH Image software indicating at least 62.5 ng of G-Rb1, -Rc and -Rd were clearly detectable individually. Three ginsenosides can be analyzed quantitatively between 0.125 and 2.0 µg.

Keywords: ginsenosides/monoclonal antibody/eastern blotting/chromatographic resolution/NIH Image software.
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