scholarly journals Oestrogenic Isoflavone Content in Natural Strains of Subterranean Clover (Trifolium subterraneum L.) from Sardinia

2006 ◽  
Vol 1 (7) ◽  
pp. 1934578X0600100 ◽  
Author(s):  
Aldo Tava ◽  
Luciano Pecetti ◽  
Alessia Bertoli ◽  
Efisio Piano
Keyword(s):  
Trifolium Pratense ◽  
Red Clover ◽  
Subterranean Clover ◽  
Trifolium Subterraneum ◽  
Biochanin A ◽  
Lower Amount ◽  
Positive Role ◽  
Isoflavone Concentration ◽  
Trifolium Pratense L ◽  
Natural Strains

A collection of subterranean clover (Trifolium subterraneum L.) genotypes from Sardinia was evaluated for the content of oestrogenic isoflavones to assess differences in total and individual compounds. Daidzein, genistein, and their respective 4′-methoxy derivatives biochanin A and formononetin, were detected in leaves and quantified by GC/FID and GC/MS methods. A large variation among genotypes for individual compounds and for the total isoflavone concentration was observed. Total isoflavones were measured in the range 0.8-13.4 mg/g fresh weight (0.3-4.5% dry matter). Daidzein was usually present in lower amount compared to the other isoflavones. Genistein and biochanin A were found in most cases in higher concentration than formononetin. Phyto-oestrogens have been shown to have both oestrogenic and anti-oestrogenic properties, and their effects entail a positive role for the human health. T. subterraneum revealed the presence of higher concentration of isoflavones compared to other sources of these compounds, e.g. soybean (Glycine max (L.) Merrill) and red clover (Trifolium pratense L.). This study provided information on the biodiversity of the natural strains from Sardinia, emphasizing the importance of these genetic resources for their variation in the amount of total and individual isoflavones.

Quantitative aspects of phytooestrogen metabolism in sheep fed on subterranean clover (Trifolium subterraneum cultivar clare) or red clover (Trifolium pratense)

1970 ◽  
Vol 21 (5) ◽  
pp. 713 ◽  
Author(s):  
DA Shutt ◽  
RH Weston ◽  
JP Hogan
Keyword(s):  
Residence Time ◽  
Trifolium Pratense ◽  
Daily Intake ◽  
Red Clover ◽  
Subterranean Clover ◽  
Trifolium Subterraneum ◽  
Biochanin A ◽  
Free Form ◽  
The Mean ◽  
High Level

Studies have been made of the digestion and metabolism in Merino wethers of the isoflavones in subterranean clover (Trtfolium subterraneurn cv. Clare) and red clover (T. pratense). The dietary intake of isoflavones with both clovers was about 9 g per day. With the subterranean clover, the isoflavones were predominantly genistein and biochanin A, and slight teat length increases in the wethers ingesting this clover indicated a low level of oestrogenicity. With the red clover formononetin represented 60% of the isoflavone present and the wethers on this diet exhibited maximal teat length increases indicating a high level of oestrogenicity. Less than 1 % of the daily intake of the isoflavones was excreted as such in the faeces and urine; hence most of these compounds were metabolized or retained in the sheep. The dietary isoflavones were found to disappear rapidly from the rumen, and it was estimated by using marker techniques that the removal of these compounds from the stomach (rumen, reticulum, omasum, and abomasum) was virtually complete. Equol (7,4'-dihydroxyisoflavan), a metabolite of formononetin, was the predominant phyto-oestrogen in the digesta and excreta when red clover was given. The excretion of 3.9 g/day of this compound, mainly in urine, was equivalent to 70% of the intake of formononetin. It was calculated that about 86% of the equol produced in the rumen was absorbed from that organ; the mean residence time for equol in the rumen was estimated to be 1.7 hr. The isoflavones were present in blood plasma mainly in conjugated forms. Equol predominated with both clovers. The levels of equol were much lower with the subterranean clover than with the red clover diet; the concentrations of the conjugated form were respectively 13 and 300-440 �g/100 ml. Equol in the free form, although not detectable with the subterranean clover, was present at 4-10 �g/100 ml with red clover. The data were considered to be consistent with the conclusion that equol accounts for most of the phyto-oestrogenic activity in sheep fed on clovers containing high levels of formononetin.

Genetic variability of principal isoflavones in red clover

2006 ◽  
Vol 86 (Special Issue) ◽  
pp. 1345-1347 ◽  
Author(s):  
Yousef A Papadopoulos ◽  
R. Tsao ◽  
K. B. McRae ◽  
A. E. Mellish ◽  
S. A. E. Fillmore
Keyword(s):  
Genetic Variability ◽  
Trifolium Pratense ◽  
Red Clover ◽  
Broad Sense ◽  
Biochanin A ◽  
Growth Stages ◽  
Broad Sense Heritability ◽  
Food Ingredients ◽  
Isoflavone Concentration ◽  
Trifolium Pratense L

Isoflavones, known for their health benefits, are abundant in red clover (Trifolium pratense L.). Total isoflavone concentrations can be 30 times that of soybean, indicating that red clover is a good source of nutraceutical and functional food ingredients. In this study, tissue samples of 13 red clover cultivars were taken at two growth stages (late-bud stage and late-flowering stage) to determine the concentration of individual isoflavones using HPLC. Individual isoflavone concentrations and total isoflavone concentration differed significantly according to red clover cultivar. We found significant genetic variability for total isoflavone concentration and individual isoflavone concentrations; these differences were not related to ploidy level (diploid vs. tetraploid). Broad-sense heritability (H = genetic variance/total variance) ranged from 0 to 83% and was influenced by isoflavone type and sampling date. The results of this study suggest that there is significant genetic variability for isoflavone concentrations among currently available red clover cultivars. Key words: Trifolium pratense L., biochanin A, formononetin, growing stages, broad-sense heritability

scholarly journals Variety, time of harvest and conditions during growing season have impact on red clover isoflavone content

2018 ◽  
Vol 27 (2) ◽  
Author(s):  
Eeva Arja Mustonen ◽  
Mikko Tuori ◽  
Päivi Kurki ◽  
Mika Isolahti ◽  
Juhani Taponen ◽  
...  
Keyword(s):  
High Performance ◽  
Trifolium Pratense ◽  
Red Clover ◽  
Weather Conditions ◽  
Biochanin A ◽  
Growing Seasons ◽  
Crop Varieties ◽  
Isoflavone Content ◽  
Trifolium Pratense L ◽  
Time Of Harvest

Red clover (Trifolium pratense L.) is the predominant legume used in northern European agriculture. Official red clover variety trials are conducted by Natural Resources Institute Finland (Luke) to determine the value of field crop varieties. The trials used for the current analysis were conducted in Luke units in southern Finland (Mikkeli) and northern Finland (Ruukki) in two consecutive years. Plant samples for isoflavone analyses were collected from four varieties grown as four replicates and harvested twice during both growing seasons. The four main isoflavones biochanin A, genistein, daidzein and formononetin were analysed using high performance liquid chromatography. Total phytoestrogen content in the varieties varied in the range of 11.2−14.8 mg g-1 dry matter (DM). The variety and the time of harvest had most effect on the isoflavone, especially formononetin, contents of red clover. A more northern growing area and challenging weather conditions were associated with increased isoflavone concentrations.

Effects of pH and aluminium on the growth of temperate pasture species. II. Growth and nodulation of legumes

1991 ◽  
Vol 42 (5) ◽  
pp. 893 ◽  
Author(s):  
DC Edmeades ◽  
FPC Blamey ◽  
CJ Asher ◽  
DG Edwards
Keyword(s):  
White Clover ◽  
Trifolium Pratense ◽  
Inorganic Nitrogen ◽  
Red Clover ◽  
Subterranean Clover ◽  
Trifolium Subterraneum ◽  
Lotus Corniculatus ◽  
Culture Solution ◽  
Solution Ph ◽  
Lotus Pedunculatus

Ten temperate pasture legumes inoculated with appropriate rhizobia were grown for 31 days in flowing solution culture. Solution ionic strength was approximately 2700 8M and contained inorganic nitrogen (150 , 8M NO3-) only at the commencement of the experiment. Solution pH was maintained at 4.5, 5.0, 5.5 and 6.0. Also, five aluminium (Al) treatments were imposed, with nominal Al concentrations of 0, 3, 6, 12 and 24 8M (2.5, 7.1, 8.3, 11.2 and 24.7 8M Al measured) at pH 4.5. Solution pH <6 . 0 markedly reduced total dry mass (TDM) in all cultivars of white clover (Trifolium repens) cvv. 'Grasslands Pitau, Huia, G18 and Tahora' and red clover (Trifolium pratense) cvv. 'Grassland Turoa and Pawera', and to a lesser extent in the two subterranean clover (Trifolium subterraneum) cvv. 'Tallarook and Woogenellup'. In contrast, solution pH had no effect on the growth of Lotus corniculatus cv. Maitland, while Lotus pedunculatus cv. Maku grew best at pH 4.5. Lotus pedunculatus cv. Maku grew best in solution where the sum of the activities of the monomeric Al species {Alm} was maintained at 5.9 8M. The growth of all other species was decreased with Al in solution, a 50% reduction in TDM being associated with c. 6 8M {Alm] for white clover and subterranean clover, and c. 3 8M in red clover and Lotus corniculatus cv. Maitland.

scholarly journals Single-Laboratory Validation of UHPLC-MS/MS Assays for Red Clover Isoflavones in Human Serum and Dietary Supplements

2020 ◽  
Vol 103 (4) ◽  
pp. 1160-1166 ◽  
Author(s):  
Ruth N Muchiri ◽  
Richard B van Breemen
Keyword(s):  
Quantitative Analysis ◽  
Human Serum ◽  
Dietary Supplements ◽  
Trifolium Pratense ◽  
Red Clover ◽  
Biochanin A ◽  
Internal Standards ◽  
Selective Reaction ◽  
Coefficients Of Variation ◽  
Trifolium Pratense L

Abstract Background Extracts of red clover (Trifolium pratense L.) containing estrogenic and pro-estrogenic isoflavones are used in dietary supplements primarily for the management of menopausal symptoms in women. Objective A UHPLC-MS/MS assay was developed and validated for the quantitative analysis of the six major red clover isoflavones in dietary supplements and in human serum in support of clinical trials. Methods Enzymatic deconjugation of isoflavone glucuronides and sulfate conjugates in human serum specimens was carried out followed by protein precipitation. Isoflavones in red clover dietary supplements were acid hydrolyzed to release aglycons from glycosides. UHPLC separations (&lt; 4 min) were combined with MS/MS using collision-induced dissociation, selective reaction monitoring and deuterated internal standards to measure biochanin A, formononetin, daidzein, genistein, irilone, and prunetin. Results The method was validated with respect to selectivity, specificity, accuracy, linearity, precision, LOD, and LOQ. The calibration curves for all analytes were linear (R2 &gt; 0.998). The mean recovery for low-, medium- and high-quality control standards ranged between 80% and 108%. The precision of the method was assessed using coefficients of variation, which were &lt;15%. Conclusions The UHPLC-MS/MS method is fast, precise, sensitive, selective, accurate, and applicable to the quantitative analysis of red clover isoflavones in different matrices. Highlights This validated UHPLC-MS/MS assay is applicable to the rapid quantitative analysis of red clover isoflavones in human serum and in dietary supplements.

Varietal and environmental variations in isoflavone concentrations in subterranean clover (Trifolium subterraneum L.)

1968 ◽  
Vol 19 (1) ◽  
pp. 15 ◽  
Author(s):  
FHW Morley ◽  
CM Francis
Keyword(s):  
Environmental Variables ◽  
Subterranean Clover ◽  
Trifolium Subterraneum ◽  
Biochanin A ◽  
Components Of Variance ◽  
Isoflavone Concentration ◽  
Environmental Variations ◽  
Breeding Selection ◽  
Low Levels ◽  
Selection For

The concentrations of formononetin, genistein, and biochanin A in leaves of 151 lines belonging to three subspecies of T. subterraneum L. and of 8 lines of T. israeliticum D. Zoh. & Katzn, were measured from plants grown in the field at Perth and Canberra, and in a glasshouse at Perth. The average concentrations did not differ significantly between locations, but did between sampling dates. The components of variance were greater for subspecies, and for varieties within subspecies, than for environmental variables other than error. Interactions were negligible, although some were greater than zero. The component correlations between concentrations of different isoflavones were generally positive where determined by environment, but tended to be negative where determined by genotype, especially that between genistein and biochanin A. Differences among subspecies In level and pattern of isoflavone concentration could be of limited value for taxonomic classification. Differences among varieties within subspecies are sufficient to promise progress in selection for low levels of individual isoflavones by conventional techniques of plant breeding. Selection for low levels of total isoflavones would be hindered, but not prevented, by the negative genotypic correlations between concentrations of genistein and biochanin A.

scholarly journals Integrated Phytochemical Analysis Based on UPLC-MS and Network Pharmacology Approaches to Explore the Quality Control Markers for the Quality Assessment of Trifolium pratense L.

Molecules ◽  
2020 ◽  
Vol 25 (17) ◽  
pp. 3787
Author(s):  
Liyu Luo ◽  
Wenya Gao ◽  
Yan Zhang ◽  
Chang Liu ◽  
Guopeng Wang ◽  
...  
Keyword(s):  
Quality Control ◽  
Quality Assessment ◽  
Chlorogenic Acid ◽  
Trifolium Pratense ◽  
Red Clover ◽  
Biochanin A ◽  
Leguminous Plant ◽  
Network Pharmacology ◽  
Trifolium Pratense L ◽  
Novel Method

Red clover consists of the overground parts and inflorescence of Trifolium pratense L., a leguminous plant belonging to the genus Trifolium. It is widely distributed worldwide and has long been used in traditional medicine. In this study, a combination approach using UPLC-MS and network pharmacology was applied to explore the quality control markers for the quality assessments of red clover. Firstly, UPLC-MS was used to identify the compounds in different parts of red clover. Twenty-eight compounds were totally identified. According to the traditional clinical efficacy of red clover, a compound-target-function network was constructed by network pharmacology to discover the main active compounds based on the identified compounds. Nine compounds of chlorogenic acid, daidzin, calycosin-7-O-β-d-glucoside, genistin, ononin, daidzein, genistein, formononetin, and biochanin A were filtrated and further confirmed in rat plasma in view of the blood-absorbed components taking effects. Finally, a novel method for simultaneously detecting the nine quality control markers was developed by UPLC-QQQ-MS in an effort to assess the quality of red clover. For all samples, the average contents of the nine compounds measured from high to low consist of formononetin, ononin, biochanin A, genistin, daidzin, calycosin-7-O-β-d-glucoside, genistein, daidzein, and chlorogenic acid. The samples from Gansu province showed the best quality in the three producing areas This study provides new strategies to explore the quality control markers and develops a novel method for the quality assessment of red clover.

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