The effect of supplementing a basal diet for dairy cows with "Soybean extract 40" (Biomedica, Prague, Czech Republic), containing 40% soybean isoflavones, on the contents of daidzein, glycitein, genistein, and equol in milk as well as fresh and mature yogurts was estimated. To determine the contents of these isoflavonoids, an efficient analytical LC-MS (TOF) technique was used. The "Soybean extract 40" used in our study contained an especially high proportion of daidzein (307gkg-1). In both milk and yogurt samples, the amounts of daidzein and its metabolite equol were significantly higher in samples obtained from cows that received the isoflavone extract-supplemented diet than from those that received the basal diet, as the precursor daidzein contributed to the increased equol concentrations. Fermentation caused significant changes in the daidzein and glycitein concentrations. With maturation, the concentrations of daidzein and equol were unaffected, while the glycitein concentration decreased significantly.

• Keywords
• Bovine milk, Daidzein (PubChem CID: 5281708), Dairy products, Equol, Equol (PubChem CID: 382975), Genistein (PubChem CID: 5280961), Glycitein (PubChem CID: 5317750), Isoflavones, Phytoestrogens, Storage,
• MeSH
• Chromatography, Liquid MeSH
• Equol analysis   MeSH
• Glycine max chemistry   MeSH
• Mass Spectrometry MeSH
• Isoflavones analysis   MeSH
• Yogurt analysis   MeSH
• Milk chemistry   MeSH
• Cattle MeSH
• Animals MeSH
• Check Tag
• Cattle MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• daidzein MeSH Browser
• Equol MeSH
• glycitein MeSH Browser
• Isoflavones MeSH

Phytoestrogens are naturally occurring nonsteroidal phenolic plant compounds that, due to their molecular structure and size, resemble vertebrate steroids estrogens. This review is focused on plant flavonoids isoflavones, which are ranked among the most estrogenic compounds. The main dietary sources of isoflavones for humans are soybean and soybean products, which contain mainly daidzein and genistein. When they are consumed, they exert estrogenic and/or antiestrogenic effects. Isoflavones are considered chemoprotective and can be used as an alternative therapy for a wide range of hormonal disorders, including several cancer types, namely breast cancer and prostate cancer, cardiovascular diseases, osteoporosis, or menopausal symptoms. On the other hand, isoflavones may also be considered endocrine disruptors with possible negative influences on the state of health in a certain part of the population or on the environment. This review deals with isoflavone classification, structure, and occurrence, with their metabolism, biological, and health effects in humans and animals, and with their utilization and potential risks.

• Keywords
• biochanin A, daidzein, equol, formononetin, genistein, glycitein, isoflavones, phytoestrogens,
• MeSH
• Equol chemistry   classification   metabolism   MeSH
• Phytoestrogens chemistry   classification   metabolism   MeSH
• Genistein chemistry   classification   metabolism   MeSH
• Isoflavones chemistry   classification   metabolism   MeSH
• Humans MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Review MeSH
• Names of Substances
• biochanin A MeSH Browser
• daidzein MeSH Browser
• Equol MeSH
• formononetin MeSH Browser
• Phytoestrogens MeSH
• Genistein MeSH
• glycitein MeSH Browser
• Isoflavones MeSH

In this research, a microbial endophytic strain obtained from the rhizosphere of the conifer Taxus baccata and designated as Streptomyces sp. AC35 (FJ001754.1 Streptomyces, GenBank) was investigated. High 16S rDNA gene sequence similarity suggests that this strain is closely related to S. odorifer. The major fatty acid profile of intracellular lipids was also carried out to further identify this strain. Atomic force microscopy and scanning acoustic microscopy were used to image our strain. Its major excreted substances were extracted, evaluated for antimicrobial activity, purified, and identified by ultraviolet-visible spectroscopy (UV-vis), liquid chromatography-mass spectrometry (LC-MS/MS) and nuclear magnetic resonance as the bioactive isoflavone aglycones-daidzein, glycitein and genistein. Batch cultivation, performed under different pH conditions, revealed enhanced production of antimycin components when the pH was stable at 7.0. Antimycins were detected by HPLC and identified by UV-vis and LC-MS/MS combined with the multiple reaction monitoring. Our results demonstrate that Streptomyces sp. AC35 might be used as a potential source of effective, pharmaceutically active compounds.

• Keywords
• Antimycin, Atomic force microscopy, Isoflavone aglycones, Streptomyces sp.,
• MeSH
• Antimycin A analogs & derivatives   metabolism   MeSH
• Genistein metabolism   MeSH
• Isoflavones biosynthesis   MeSH
• Streptomyces chemistry   genetics   metabolism   ultrastructure   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Antimycin A MeSH
• daidzein MeSH Browser
• Genistein MeSH
• glycitein MeSH Browser
• Isoflavones MeSH

The aim of the study was to determine the ruminal degradability of dry matter (DM), daidzein, genistein, glycitein and total isoflavones in ground full-fat soybean (GFFS) and solvent-extracted soybean meal (SSBM) using the in sacco method. The experiment was carried out in three replications on ruminally cannulated sheep that were fed twice a day with a diet consisted of hay and supplemental mixture (6:4, DM basis). The nylon bags with 2 g feed samples ground to 2 mm were incubated in the rumen for 0, 2, 4, 8, 16 and 24 h. The effective degradability (ED) of DM, daidzein, genistein, glycitein and total isoflavones was calculated at outflow rate of 0.06 h. The ED of DM in GFFS was 77.8% and was higher than in SSBM being 71.8% (p < 0.001). The ED of daidzein (96.8%) and genistein (93.6%) was higher for SSBM compared with GFFS (93.9% and 92.8%, p < 0.001 and p = 0.003, respectively) while ED of glycitein was lower for SSBM than for GFFS (75.5 and 81.7%, respectively, p < 0.001). All isoflavones in the incubations were extensively degraded in the rumen, and regardless of dietary source, they were almost completely degraded after 16 h of incubation. Further, the disappearance patterns, that is the functions describing the time courses of the analyte disappearance, were assessed. The disappearance patterns of daidzein, genistein, glycitein and total isoflavones were similar and showed greater disappearance of mentioned isoflavones from SSBM compared to GFFS (p < 0.001 for daidzein, genistein and total isoflavones and p = 0.002 for glycitein). The study provides knowledge on the effect of processing on degradability of isoflavones in rumen that can be used to clarify the interrelationship between isoflavones and rumen microbiota.

• Keywords
• daidzein, genistein, glycitein, ground full-fat soybean, solvent-extracted soybean meal, total isoflavones,
• MeSH
• Rumen MeSH
• Diet veterinary   MeSH
• Glycine max * MeSH
• Isoflavones * MeSH
• Sheep MeSH
• Pilot Projects MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Isoflavones * MeSH

1. The possibility of interaction of isoflavonoids with concomitantly taken drugs to determined isoflavonoids safety was studied. Inhibition of nine forms of cytochrome P450 (CYP3A4, CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C19, CYP2C9, CYP2D6 and CYP2E1) by 12 isoflavonoids (daidzein, genistein, biochanin A, formononetin, glycitein, equol and six glucosides, daidzin, puerarin, genistin, sissotrin, ononin and glycitin) was studied systematically. 2. The most potent inhibitors were genistein and daidzein inhibiting noncompetitively the CYP2C9 with Ki of 35.95 ± 6.96 and 60.56 ± 3.53 μmol/l and CYP3A4 (inhibited by genistein with Ki of 23.25 ± 5.85 μmol/l also by a noncompetitive mechanism). Potent inhibition of CYP3A4 was observed also with biochanin A (Ki of 57.69 ± 2.36 μmol/l) and equol (Ki of 38.47 ± 2.32 μmol/l). 3. Genistein and daidzein inhibit noncompetitively CYP3A4 and CYP2C9. With plasma levels in micromolar range, a clinically important interaction with concomitantly taken drugs does not seem to be probable.

• Keywords
• Daidzein, drug interactions, genistein, inhibition studies,
• MeSH
• Cytochrome P-450 CYP1A2 MeSH
• Cytochrome P-450 CYP2C19 MeSH
• Glucosides MeSH
• Isoflavones metabolism   MeSH
• Microsomes, Liver enzymology   MeSH
• Liver enzymology   MeSH
• Drug Interactions MeSH
• Humans MeSH
• Cytochrome P-450 Enzyme System metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• calycosin-7-O-beta-D-glucoside MeSH Browser
• CYP1A2 protein, human MeSH Browser
• Cytochrome P-450 CYP1A2 MeSH
• Cytochrome P-450 CYP2C19 MeSH
• formononetin MeSH Browser
• genistin MeSH Browser
• Glucosides MeSH
• glycitein MeSH Browser
• glycitin MeSH Browser
• Isoflavones MeSH
• puerarin MeSH Browser
• Cytochrome P-450 Enzyme System MeSH

Complete separation of aglycones and glucosides of selected isoflavones (genistin, genistein, daidzin, daidzein, glycitin, glycitein, ononin, sissotrin, formononetin, and biochanin A) was possible in 1.5 min using an ultrahigh-pressure liquid chromatography (U-HPLC) on a different particular chemically modified stationary phases with a particle size under 2 microm. In addition, selected separation conditions for simultaneous determination of isoflavones together with a group of phenolic acids (gallic, protocatechuic, p-hydroxybenzoic, vanillic, caffeic, syringic, p-coumaric, ferulic, and sinapic acid) allowed separation of all 19 compounds in 1.9 min. Separations were conducted on a non-polar reversed phase (C(18)) and also on more polar phases with cyanopropyl or phenyl groups using a gradient elution with a mobile phase consisting of 0.3% aqueous acetic acid and methanol. Chromatographic peaks were characterised using parameters such as resolution, symmetry, selectivity, etc. Individual substances were identified and quantified using UV-vis diode array detector at wavelength 270 nm. Limits of detection (3S/N) were in the range 200-400 pg ml(-1). Proposed U-HPLC technique was used for separation of isoflavones and phenolic acids in samples of plant materials (Trifolium pratense, Glycine max, Pisum sativum and Ononis spinosa) after acid hydrolysis of the samples and modified Soxhlet extraction.

• MeSH
• Genistein chemistry   isolation & purification   MeSH
• Glycine max chemistry   MeSH
• Pisum sativum chemistry   MeSH
• Hydroxybenzoates chemistry   isolation & purification   MeSH
• Isoflavones chemistry   isolation & purification   MeSH
• Gallic Acid analogs & derivatives   chemistry   isolation & purification   MeSH
• Vanillic Acid chemistry   isolation & purification   MeSH
• Caffeic Acids chemistry   isolation & purification   MeSH
• Coumaric Acids chemistry   isolation & purification   MeSH
• Molecular Structure MeSH
• Propionates MeSH
• Plant Extracts chemistry   isolation & purification   MeSH
• Trifolium chemistry   MeSH
• Chromatography, High Pressure Liquid methods   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• biochanin A MeSH Browser
• caffeic acid MeSH Browser
• daidzein MeSH Browser
• daidzin MeSH Browser
• ferulic acid MeSH Browser
• formononetin MeSH Browser
• Genistein MeSH
• genistin MeSH Browser
• glycitein MeSH Browser
• glycitin MeSH Browser
• Hydroxybenzoates MeSH
• Isoflavones MeSH
• Gallic Acid MeSH
• Vanillic Acid MeSH
• Caffeic Acids MeSH
• Coumaric Acids MeSH
• p-coumaric acid MeSH Browser
• phenolic acid MeSH Browser
• Propionates MeSH
• protocatechuic acid MeSH Browser
• Plant Extracts MeSH
• sinapinic acid MeSH Browser
• syringic acid MeSH Browser

A direct analysis in real-time (DART) ion source coupled to a high-resolution orbitrap mass spectrometer was used for the quantitative analysis of isoflavones isolated from soybeans. For the isolation of genistein, daidzein, glycitein, and their respective acetyl, malonyl, and glucoside forms, an extraction employing 80% aqueous MeOH enhanced by sonication was used. As far as the total isoflavones (expressed as aglycones) were to be determined, an acid hydrolysis with 80% aqueous EtOH and refluxing had to be employed, while in the latter case a good agreement of the results with the data generated by the UHPLC-orbitrap MS method was achieved, in the case of the analysis of non-hydrolyzed extracts, some overestimation of the results as compared with those generated by UHPLC-orbitrap MS was observed. A careful investigation of this phenomenon showed that the free aglycones originated from the conjugated forms of isoflavones in the DART ion source, thus contributing significantly to the "free" genistein/daidzein/glycitein signals during the DART analysis. Good recoveries (95-102%) and repeatabilities (RSD: 7-15%) were obtained at the spiking levels of 0.5, 1, and 0.05 g/kg, for daidzein, genistein, and glycitein, respectively. The limits of detection estimated for the respective analytes were 5 mg/kg.

• MeSH
• Time Factors MeSH
• Glycine max chemistry   MeSH
• Spectrometry, Mass, Electrospray Ionization MeSH
• Isoflavones analysis   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Isoflavones MeSH

The dietary intake of flavonoids seems to be inversely related to cardiovascular mortality. The consumption of isoflavonoids is increasing in the general population, especially due to the use of food supplements and a variety of isoflavonoid-rich foods. However, detailed studies on the vascular influence of individual pure isoflavonoids are mostly missing. For this study, 16 isoflavonoids were initially screened for their vasorelaxant properties on rat aortas. The 2 most potent of them, biochanin A and glycitein, were further tested for the mechanism of action on porcine coronary arteries. They both induced an endothelium independent vascular relaxation, with EC50 below 6 and 17 µM, respectively. Biochanin A, but not glycitein, was able to block the vasoconstriction caused by KCl, CaCl2, serotonin, and U46619 in a dose-dependent manner. Another series of experiments suggested that the major mechanism of action of biochanin A was the inhibition of L-type calcium channels. Moreover, biochanin A in relatively small concentrations (2 - 4 µM) interfered with the cGMP, but not cAMP, pathway in isolated coronary arteries. These results indicate that some isoflavonoids, in particular biochanin A, are able to have vasodilatory effects in micromolar concentrations, which is of potential clinical interest for the management of cardiovascular pathologies.

• MeSH
• Endothelium, Vascular MeSH
• Genistein MeSH
• Isoflavones * MeSH
• Coronary Vessels MeSH
• Rats MeSH
• Humans MeSH
• Swine MeSH
• Calcium MeSH
• Calcium Channels * MeSH
• Vasodilation MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• biochanin A MeSH Browser
• Genistein MeSH
• Isoflavones * MeSH
• Calcium MeSH
• Calcium Channels * MeSH

An efficient method of modifier addition for supercritical fluid extraction (SFE) of polar isoflavones was developed and yielded extraordinarily high recoveries. To find the optimal extraction conditions, a temperature and pressure optimization and modifier impact study was performed in naturally contaminated and spiked samples. Ultra-fast high-performance liquid chromatography/mass spectrometry (HPLC/MS) was used for the determination of isoflavones on an Atlantis dC18 high-speed reversed phase chromatographic column (20 x 2.1 mm, 3 microm particle size). A newly elaborated supercritical fluid extraction (SFE) procedure allowed more accurate (< 5%) and precise (< 4-7%) determination of isoflavones in biological materials. The HPLC/MS method significantly reduced analysis time with simultaneous improvement of sensitivity and detection limits. The on-column limits of detection LOD (S/N = 3) for isoflavone glycosides (daidzin, genistin, glycitin, ononin, and sissotrin) were 1.3-3.6 fmol and 0.2-1.0 fmol for aglycones (daidzein, glycitein, genistein, formononetin, and biochanin A), respectively.

• MeSH
• Food Analysis methods   MeSH
• Glycine max chemistry   MeSH
• Glycosides chemistry   isolation & purification   MeSH
• Mass Spectrometry methods   MeSH
• Isoflavones chemistry   isolation & purification   MeSH
• Trifolium chemistry   MeSH
• Chromatography, High Pressure Liquid methods   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Validation Study MeSH
• Names of Substances
• Glycosides MeSH
• Isoflavones MeSH

Several types of compounds with immunoreactivity similar to isoflavonoids were detected in water: ethanol extracts of leaves of Fortunella obovata Hort. ex Tanaka, Murraya paniculata Jack. and four Citrus species, namely C. aurantium L, C. grandis Osbeck, C. limonia Osbeck., and C. sinensis Osbeck (Rutaceae). The chromatographic mobilities of the immunoreactive substances were compared with those of authentic standards, revealing a spectrum of isoflavonoid metabolites in all plants studied. Aglycones as well as glycosides were recognized, namely daidzin, genistin, daidzein, genistein, formononetin, biochanin A, prunetin, and several incompletely characterized isoflavonoids. A subsequent HPLC-MS study verified the identities of the main immunoreactive isoflavonoids and established the identities of several others, viz. glycitein, glycitin, ononin and sissotrin, including the malonylated and acetylated isoflavonoid glucosides. The estimated content of the individual immunoreactive entities ranged from a few microg to about 2 mg/kg (dry weight). It is concluded that the isoflavonoid metabolic pathway is present throughout the Rutaceae family.

• MeSH
• Chromatography, Thin Layer MeSH
• Genistein chemistry   metabolism   MeSH
• Mass Spectrometry MeSH
• Isoflavones analysis   chemistry   metabolism   MeSH
• Plant Leaves chemistry   metabolism   MeSH
• Radioimmunoassay MeSH
• Rutaceae chemistry   metabolism   MeSH
• Chromatography, High Pressure Liquid MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• daidzein MeSH Browser
• Genistein MeSH
• Isoflavones MeSH