ENZYME-IMMUNOASSAY FOR THE QUANTITATIVE-DETERMINATION OF GALANTHAMINE

An enzyme immunoassay for the quantitation of fmol amounts of the ther apeutically important Amaryllidaceae alkaloid, galanthamine, was estab lished. The antiserum was raised against a conjugate of galanthamine-2 -O-hemisuccinate-bovine serum albumin. The antibodies used were isolat ed and purified by Rivanol treatment with subsequent (NH4)2SO4 precipi tation. The measuring range of the assay extends from 2 to 100 pg of g alanthamine, and as little as 3.5 fmol may be detected. The antibodies are highly specific for galanthamine, showing no cross reactivity wit h several Amaryllidaceae alkaloids. This assay allows the rapid, sensi tive and precise quantitation of galanthamine in unpurified plant extr acts as well as biological fluids. The galanthamine content in a varie ty of herbarium material as well as the frequency distribution of gala nthamine in 1000 Leucojum aestivum plants from various origins in Sout h Bulgaria have been investigated. The preliminary results demonstrate that the galanthamine-specific enzyme immunoassay can be a useful too l in medicinal plant breeding, for screening programs, as well as for taxonomic and pharmacokinetic studies.