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Glycobiology Advance Access originally published online on June 22, 2005
Glycobiology 2005 15(10):1061-1066; 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

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

Osamu Morinaga2, Noriko Fukuda3, Hiroyuki Tanaka3 and Yukihiro Shoyama1,3

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

1 To whom correspondence should be addressed; e-mail: shoyama{at}phar.kyushu-u.ac.jp

Received on May 12, 2005; revised on June 1, 2005; accepted on June 14, 2005

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), G-Rc, and G-Rd in crude extracts of various ginsengs. The 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 the first antibody. Finally a substrate reacted to the enzyme and gave staining. The stained membrane was scanned, and spots were analyzed quantitatively using NIH Image software. At least 62.5 ng of G-Rb1, G-Rc, and G-Rd were clearly detectable individually. Three ginsenosides can be analyzed quantitatively between 0.125 and 2.0 µg.

Key words: chromatographic resolution / eastern blotting / ginsenosides / monoclonal antibody / NIH Image software


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