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Advances in isoflavone profile characterisation using matrix solid-phase dispersion coupled to HPLC/DAD in medicago species

Publication . Visnevschi-Necrasov, Tatiana; Barreira, João C.M.; Cunha, Sara C.; Pereira, Graça; Nunes, Eugénia; Oliveira, Beatriz

Analytical methods used in phytochemistry analysis are limited by the sample preparation step, which should ideally be fast, accurate, ecofriendly and achievable using low quantities of the sample. Matrix solid-phase dispersion (MSPD) may be a good alternative for combining extraction and purification procedures, thereby reducing the indicated limitations. Objective-Applying an MSPD extraction procedure coupled to high-performance liquid chromatography diode-array detection (HPLC/DAD) as an alternative methodology to evaluate isoflavone profiles. Methods-Isoflavone profiles were determined for the leaves of nine species of Medicago in the late flower phenological stage (one or more nodes with 50% open flowers, no seed pods). Extraction was performed following MSPD, and isoflavone profiles were characterised using HPLC/DAD. The quantified amounts were compared with previous results in different species commonly recognised as good sources of isoflavones. Results-Formononetin was the major isoflavone in most species, except M. polymorpha and M. truncatula. The isoflavone amounts were significantly different among the assayed species, with M. orbicularis and M. arabica as the major isoflavone sources, while M. rigidula presented the lowest contents. Furthermore, the detected differences allow electing the best species as a primary source of a specific isoflavone. Conclusion-The MSPD allowed good extraction efficiency, reproducibility and recovery. Some of the species showed relevant isoflavone contents, even when compared with acknowledged plant sources such as soy or red clover. To the best of our knowledge the results presented are reported for the first time in these species.

2015Journal article

Open access

Phytochemical profiling of underexploited Fabaceae species: Insights on the ontogenic and phylogenetic effects over isoflavone levels

Publication . Barreira, João C.M.; Visnevschi-Necrasov, Tatiana; Pereira, Graça; Nunes, Eugénia; Oliveira, Beatriz

There is an increasing trend towards finding alternative sources of valued phytochemicals due to their diverse potentialities in food industry and pharmaceutical applications. Phenolic compounds, in particular, have been the focus of several profiling reports, but isoflavones characterization has been studied in fewer cases and in a very limited group of plant species. Despite their acknowledged bioactivity, there's actually a strict number of plants validated for their isoflavones contents. In a previous report, we have identified nine Leguminosae species (from genera Biserrula, Lotus, Ornithopus and Scorpiurus) as potential alternative sources of these phenolic compounds. However, the isoflavone profiles are highly modulated by the ontogenic stage. Therefore, the present study was conducted in the same Leguminosae species, but harvested at three sequential vegetative development stages: vegetative elongation, late bud and late flowering, with the main purpose of assessing the evolution of isoflavones content throughout the plant development. In general, the plant species from Biserrula and Lotus genera showed the highest potential as new natural sources of isoflavones, especially owing their high levels of biochanin A. Independently of the plant species, it was possible to identify the phenologic stages where each of the quantified isoflavones is maximized. These findings are useful to predict isoflavone yields according to harvesting time, validating the potential use of the studied plants in innovative food formulations.

2017Journal article

Open access

Medicago spp. as potential sources of bioactive isoflavones: Characterization according to phylogenetic and phenologic factors

Publication . Barreira, João C.M.; Visnevschi-Necrasov, Tatiana; Nunes, Eugénia; Cunha, Sara C.; Pereira, Graça; Oliveira, Beatriz

A high variety of plant species are often proposed as potential natural sources of specific bioactive components, with emphasis in phenolic compounds. However, the ability to produce a determined phytochemical might be variable, even among species with close phylogeny. Furthermore, the metabolic dynamics vary greatly according to phenologic factors. Herein, it was verified whether isoflavone production in Medicago spp. is more associated with phylogenetic or phenologic determinants, to define the optimal productive conditions. Isoflavone profiles were characterized in field-grown Medicago species in three phenologic stages. Isoflavones were extracted by matrix solid-phase dispersion method and analyzed using high-performance liquid chromatography coupled with a diode-array detector. The obtained data were evaluated by a generalized linear model (GLM) and linear discriminant analysis (LDA). Formononetin, genistein and irilone were the most abundant isoflavones, reaching values higher than those present in acknowledged plant sources like soy or red clover. Outputs from GLM and LDA indicate that the phylogenetic factors are the most defining criteria. This study promotes Medicago spp. as potential isoflavone sources, particularly because the effects of these compounds are highly dependent on their type and concentration, with potential application as foodstuff, feedstuff, or in the nutraceutical and pharmaceutica l industry.

2015Journal article

Open access