Chromatographic resolution of glucosidic compounds, ginsenosides on polyethersulphone membrane, and its application to the quantitative immunoassay for ginseng saponins.
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 microg.
Morinaga
Osamu
O
Innovation Plaza Fukuoka, Japan Science and Technology Agency, 3-8-34 Momochihama, Fukuoka 814-0001, Japan.
Fukuda
Noriko
N
Tanaka
Hiroyuki
H
Shoyama
Yukihiro
Y
D000911:Antibodies, Monoclonal; D002851:Chromatography, High Pressure Liquid; D036145:Ginsenosides / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification*; D007118:Immunoassay; D008567:Membranes, Artificial; D005894:Panax / Q000737:chemistry*; D010936:Plant Extracts / Q000737:chemistry; D011108:Polymers; D013450:Sulfones
D000911:Antibodies, Monoclonal; D036145:Ginsenosides; D008567:Membranes, Artificial; D010936:Plant Extracts; D011108:Polymers; D013450:Sulfones; C044462:ginsenoside Rc; C022840:polyether sulfone; C442759:ginsenoside Rb1; C049863:ginsenoside Rd