Enzyme-linked immunosorbent assay for the determination of isoflavones in alimentary important plants

Published on Feb 20, 2018in Czech Journal of Food Sciences0.932
· DOI :10.17221/10659-CJFS
Michaela Vitkova5
Estimated H-index: 5
Zuzana Mackova4
Estimated H-index: 4
+ 1 AuthorsOldřich Lapčík14
Estimated H-index: 14
The development of polyclonal antibody-based enzyme-linked immunosorbent assays (ELISA) for the determination of individual isoflavones, i.e. daidzein, genistein and biochanin and their homologues, is presented in this work. Isoflavone conjugates with bovine serum albumin were used as immunogens, coupled at the position C 7 and C 4' via a carboxy methyl spacer. The developed ELISAs are highly specific, I 5 0 values of the standard curves range between 0.3-1.2 ng/ml. The cross reactivities to other isoflavones are in acceptable range and the interference of non-isoflavonoid molecules is negligible. The immunoassays have been used for monitoring of changes in isoflavone levels in alimentary important plants, such as Medicago sativa, during germination; and during different vegetation stages of the Rutaceae family plants.
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#1Khaled Abdulmanea (Institute of Chemical Technology in Prague)H-Index: 1
#2Elena A. Prokudina (Institute of Chemical Technology in Prague)H-Index: 2
Last. Oldřich Lapčík (Institute of Chemical Technology in Prague)H-Index: 14
view all 7 authors...
Ethanolic extracts of 23 species of the Apiaceae family were analyzed for their flavonoid and isoflavonoid content, using the following methods: high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS); semi-preparative HPLC with Enzyme Linked Immunosorbent Assay (HPLC-ELISA); and immunoaffinity extraction (IAE). Rutin and quercetin-3-glucoside were the most abundant flavonoid glycosides and the most abundant flavonoid aglycone was apigenin. The isoflavonoids daidzein, gen...
13 CitationsSource
#1Radka KoblovskáH-Index: 5
#2Zuzana MackovaH-Index: 4
Last. Oldřich LapčíkH-Index: 14
view all 6 authors...
Enzyme-linked immunosorbent assays in combination with semi-preparative high-performance liquid chromatography (HPLC) and analytical HPLC with mass spectroscopy in the selective ion monitoring mode were used for the determination of selected isoflavones, daidzein, genistein, biochanin A and their homologues, in 20 representatives of the Rutaceae family. Species belonging to five genera were studied, namely Citrus, Fortunella, Poncirus, Ruta and Severinia. The enzyme immunoassays used were based ...
12 CitationsSource