Enzymová imunoanalyza pro stanovení isoflavonoid˚U

Published on Jan 1, 2004in Chemicke Listy0.39
Michaela Vitkova5
Estimated H-index: 5
Zuzana Mackova4
Estimated H-index: 4
+ 1 AuthorsOldrich Lapcik5
Estimated H-index: 5
Competitive enzyme-linked immunosorbent assays (ELISA) for the determination of isoflavones daidzein, genistein and biochanin were developed. Isoflavone conjugates coupled at positions C7 and C4' via a carboxymethyl spacer to bovine serum albumin were used as immunogens and also as coating antigens. Assay conditions, including concentrations of antisera and coating antigens, were optimized and the effect of reaction time, various blocking agents and organic solvents was investigated. The developed ELISA assays are highly sensitive, the I 5 0 values of standard curves ranging between 15-60 pg/well, i.e. 0.3-1.2 ng.ml - 1 . Cross reactivities to other isoflavones are in an acceptable range and interference of non-isoflavonoid molecules is negligible. The immunoassays have been used in phytochemical research for monitoring isoflavone levels in various plant families. We expect further use of the assays in analysis of food, body fluids, cell culture media and other biological materials.
Cited By6
#1L. SosvorováH-Index: 1
#2Petra LankováH-Index: 2
Last. Oldřich LapčíkH-Index: 14
view all 6 authors...
Sosvorova L., Lankova P., Bicikova M., Prokudina E.A., Al Malarik N., Lapcik O. (2011): ELISA for free S-equol in human urine. Czech J. Food Sci., 29: 57–64. The popularity of phytoestrogen food supplements of plant origin steadily increases. They are used to solve the problems related to the climacterium. Their efficiency depends on woman‘s capability to metabolise the active components – isoflavonoids. Women able to convert daidzein to S-equol are supposed to get a greater benefit from these s...
2 CitationsSource
#2Petra LankováH-Index: 2
Last. Oldřich LapčíkH-Index: 14
view all 4 authors...
Plants of the Amaryllidaceae family are known as producers of biologically active alkaloids. Besides these a variety of flavonoids, including flavones, chalcones and chromones, have been detected in the Amaryllidaceous plants. In this study, we have analysed 16 representatives of the family for the presence of isoflavonoids. The water/ethanolic extracts were analysed with HPLC-ESI-MS both without any pre-treatment and after immunoaffinity chromatography as a clean-up step. Four individual immuno...
7 CitationsSource
#1Elena A. Prokudina (Institute of Chemical Technology in Prague)H-Index: 2
#1E.A. ProkudinaH-Index: 2
Last. Oldrich Lapcik (Institute of Chemical Technology in Prague)H-Index: 5
view all 5 authors...
The majority of currently known isoflavonoids has been detected in and isolated from the Leguminosae; however, their presence has also been reported in numerous non-legumes. Detection of these secondary metabolites in non-legumes is impeded by the complex matrix of plant material and low concentration of the analytes. In this study, we present a combination of immunoaffinity chromatography (IAC) and HPLC–ESI-MS as a new tool for the detection of isoflavonoids. Immunoglobulin fractions of polyclo...
10 CitationsSource
#1Jan Vacek (H.I., S.I.: University of Agriculture, Faisalabad)H-Index: 30
#2Borivoj Klejdus (H.I., S.I.: University of Agriculture, Faisalabad)H-Index: 18
Last. Vlastimil KubanH-Index: 20
view all 4 authors...
Isoflavones are natural substances which elicit a number of physiological effects in living organisms. The substances are synthesized in plant tissues as protective agents against biotic stress (i. e. bacterial infection). Isoflavones are also an important dietary constituent in human nutrition. Modern trends in studies of isoflavones in plant materials and foodstuffs and procedures for chemical analyses of isoflavones in human body fluids and plant tissues are discussed in this review. Highly e...
128 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
#1Oldřich LapčíkH-Index: 14
#1Oldrich Lapcik (Institute of Chemical Technology in Prague)H-Index: 5
Last. Bořivoj Klejdus (H.I., S.I.: University of Agriculture, Faisalabad)H-Index: 36
view all 6 authors...
Extracts from Arabidopsis thaliana leaves and inflorescence stalks and from Lepidium sativa (Brassicaceae) seedlings were analysed by HPLC-MS-SIM and by five isoflavonoid-specific ELISA methods after the HPLC fractionation of samples, in order to determine presence of isoflavonoids. Individual ELISAs were specific for daidzein, genistein, biochanin A and for their derivatives substituted either at the 4′- or at the 7- positions. Both analytical approaches revealed homologous spectra of isoflavon...
18 CitationsSource