Lillian Barros

Antioxidants in Wild Mushrooms

Maintenance of equilibrium between free radical production and antioxidant defences (enzymatic and non enzymatic) is an essential condition for normal organism functioning. When this equilibrium has a tendency for the production of free radicals we say that the organism is in oxidative stress. In this situation, excess free radicals may damage cellular lipids, proteins and DNA, affecting normal function and leading to various diseases. In aerobic organisms, the free radicals are constantly produced during the normal cellular metabolism, mainly in the form of Reactive Oxygen Species (ROS) and Reactive Nitrogen Species (RNS). Exposition of the organism to free radicals has led to the development of endogenous defence mechanisms to eliminate them. These defences were the response of evolution to the inevitability of ROS production in aerobic conditions. Natural products with antioxidant activity may help the endogenous defence system. In this perspective the antioxidants present in the diet assume a major importance as possible protector agents reducing oxidative damage. Particularly, the antioxidant properties of wild mushrooms have been extensively studied by our research group and by others, and many antioxidant compounds extracted from these sources have been identified, such as phenolic compounds, tocopherols, ascorbic acid, and carotenoids. We will review the compounds identified so far in mushrooms, as well as the mechanism of action involved in their antioxidant properties. Wild mushrooms might be used directly in diet and promote health, taking advantage of the additive and synergistic effects of all the bioactive compounds present.

Wild and commercial mushrooms as source of nutrients and nutraceuticals

In order to promote the use of mushrooms as source of nutrients and nutraceuticals, several experiments were performed in wild and commercial species. The analysis of nutrients included determination of proteins, fats, ash, and carbohydrates, particularly sugars by HPLC-RI. The analysis of nutraceuticals included determination of fatty acids by GC-FID, and other phytochemicals such as tocopherols, by HPLC-fluorescence, and phenolics, flavonoids, carotenoids and ascorbic acid, by spectrophotometer techniques. The antimicrobial properties of the mushrooms were also screened against fungi, Gram positive and Gram negative bacteria. The wild mushroom species proved to be less energetic than the commercial sp., containing higher contents of protein and lower fat concentrations. In general, commercial species seem to have higher concentrations of sugars, while wild sp. contained lower values of MUFA but also higher contents of PUFA. alpha-Tocopherol was detected in higher amounts in the wild species, while gamma-tocopherol was not found in these species. Wild mushrooms revealed a higher content of phenols but a lower content of ascorbic acid, than commercial mushrooms. There were no differences between the antimicrobial properties of wild and commercial species. The ongoing research will lead to a new generation of foods, and will certainly promote their nutritional and medicinal use.

Chemical composition and nutritional value of the most widely appreciated cultivated mushrooms: an inter-species comparative study.

Herein, it was reported and compared the chemical composition and nutritional value of the most consumed species as fresh cultivated mushrooms: Agaricus bisporus (white and brown mushrooms), Pleurotus ostreatus (oyster mushroom), Pleurotus eryngii (King oyster mushroom), Lentinula edodes (Shiitake) and Flammulina velutipes (Golden needle mushroom). Shiitake revealed the highest levels of macronutrients, unless proteins, as also the highest sugars, tocopherols and PUFA levels, and the lowest SFA content. White and brown mushrooms showed similar macronutrients composition, as also similar values of total sugars, MUFA, PUFA and total tocopherols. Oyster and king oyster mushrooms gave the highest MUFA contents with similar contents in PUFA, MUFA and SFA in both samples. They also revealed similar moisture, ash, carbohydrates and energy values. This study contributes to the elaboration of nutritional databases of the most consumed fungi species worldwide, allowing comparison between them. Moreover it was reported that cultivated and the wild samples of the same species have different chemical composition, including sugars, fatty acids and tocopherols profiles.

Chemical composition and biological properties of portuguese wild mushrooms: a comprehensive study

The chemical composition and biological properties of Portuguese wild mushrooms ( Cantharellus cibarius, Hypholoma fasciculare, Lepista nuda, Lycoperdon molle, Lycoperdon perlatum, Ramaria botrytis, Tricholoma acerbum) were evaluated in order to assess these products as sources of nutrients and nutraceuticals. The analyzed mushrooms contain very useful phytochemicals such as phenolics, tocopherols, ascorbic acid, and carotenoids. All of the species proved to have antioxidant activity (measured by four different methods), being more significant for R. botrytis (EC 50 values < 1 mg/mL). Lycoperdon species were resistant to all of the tested microorganisms, and the other samples revealed antimicrobial activity selectively against Gram-positive bacteria, with very low minimal inhibitory concentration, in some cases, even lower than the standard. The combination of bioactive compounds and rich nutritional composition (high contents in protein and carbohydrates, low content in fat with the precious contribution of unsaturated fatty acids and the absence of trans fatty acids) in the mushroom makes it a very special food.

Phenolic acids determination by HPLC-DAD-ESI/MS in sixteen different portuguese wild mushrooms species

Analysis of phenolic compounds in sixteen Portuguese wild mushrooms species has been carried out by high-performance liquid chromatography coupled to photodiode array detector and mass spectrometer (HPLC-DAD-ESI/MS). No flavonoids were detected in the analysed samples, but diverse phenolic acids namely protocatechuic, p-hydroxybenzoic and p-coumaric acids, and two vanillic acid isomers were found and quantified. A related non-phenolic compound, cinnamic acid, was also detected in some samples, being the only compound found in Cantharellus cibarius (14.97 mg/kg, dry matter), Lycoperdon perlatum (14.36 mg/kg) and Macrolepiota procera (21.53 mg/kg). p-Hydroxybenzoic acid was found in the majority of the samples, being the most abundant compound in Agaricus silvicola (238.7 mg/kg). Ramaria botrytis showed the highest phenolic acids concentration (356.7 mg/kg) due to the significant contribution of protocatechuic acid (342.7 mg/kg).

Targeting excessive free radicals with peels and juices of citrus fruits: Grapefruit, lemon, lime and orange

A comparative study between the antioxidant properties of peel (flavedo and albedo) and juice of some commercially grown citrus fruit (Rutaceae), grapefruit (Citrus paradisi), lemon (Citrus limon), lime (Citrusxaurantiifolia) and sweet orange (Citrus sinensis) was performed. Different in vitro assays were applied to the volatile and polar fractions of peels and to crude and polar fraction of juices: 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity, reducing power and inhibition of lipid peroxidation using beta-carotene-linoleate model system in liposomes and thiobarbituric acid reactive substances (TBARS) assay in brain homogenates. Reducing sugars and phenolics were the main antioxidant compounds found in all the extracts. Peels polar fractions revealed the highest contents in phenolics, flavonoids, ascorbic acid, carotenoids and reducing sugars, which certainly contribute to the highest antioxidant potential found in these fractions. Peels volatile fractions were clearly separated using discriminant analysis, which is in agreement with their lowest antioxidant potential.

Antioxidant properties and phenolic profile of the most widely appreciated cultivated mushrooms: a comparative study between in vivo and in vitro samples.

The present study reports a comparison of the antioxidant properties and phenolic profile of the most consumed species as fresh cultivated mushrooms and their mycelia produced in vitro: Agaricus bisporus (white and brown), Pleurotus ostreatus (oyster), Pleurotus eryngii (king oyster) and Lentinula edodes (shiitake). The antioxidant activity was evaluated through reducing power (Folin-Ciocalteu and Ferricyanide/Prussian blue assays), free radical scavenging activity (DPPH assay) and lipid peroxidation inhibition (β-carotene/linoleate and TBARS assays). The analysis of phenolic compounds was performed by HPLC/PAD. The mushroom species with the highest antioxidant potential was Agaricus bispous (brown). However, concerning to the species obtained in vitro, it was L. edodes that demonstrate the highest reducing power. Generally, in vivo samples revealed higher antioxidant properties than their mycelia obtained by in vitro techniques. About the phenolic compounds researched, they were detected both in mushrooms and mycelia without any particular abundance. Results showed that there is no correlation between the studied commercial mushrooms and the corresponding mycelia obtained in vitro. Nevertheless, this study contributes to the rise of data relatively to the species consumed as fresh mushrooms and the possibility of their in vitro production as a source of bioactive compounds.

Leaves, flowers, immature fruits and leafy flowered stems of Malva sylvestris: A comparative study of the nutraceutical potential and composition

Malva sylvestris is widely used in Mediterranean and European traditional medicine and ethnoveterinary for the treatment of external and internal inflammation, as well as injuries. Moreover, its use is not only limited to therapeutic purposes; but also the species is locally regarded as a food wild herb. Considering that antioxidants and free radical scavengers can exert also an anti-inflammatory effect, the extracts of different parts of the medicinal/edible plant M. sylvestris (leaves, flowers, immature fruits and leafy flowered stems) were compared for their nutraceutical potential (antioxidant properties) and chemical composition. Particularly, mallow leaves revealed very strong antioxidant properties including radical-scavenging activity (EC(50)=0.43 mg/mL), reducing power (0.07 mg/mL) and lipid peroxidation inhibition in lipossomes (0.04 mg/mL) and brain cells homogenates (0.09 mg/mL). This part of the plant is also the richest in nutraceuticals such as powerful antioxidants (phenols, flavonoids, carotenoids, and tocopherols), unsaturated fatty acids (e.g. alpha-linolenic acid), and minerals measured in ash content.

Characterization of phenolic compounds in flowers of wild medicinal plants from Northeastern Portugal.

Crataegus monogyna, Cytisus multiflorus, Malva sylvestris and Sambucus nigra have been used as important medicinal plants in the Iberian Peninsula since a long time ago, and are claimed to have various health benefits. This study aimed to determine the phenolic profile and composition of wild medicinal flowers of those species. The analysis was performed by HPLC-DAD-ESI/MS. Flavonoids, and particularly flavonols and flavones, were the main groups in almost all the studied samples. C. multiflorus sample gave the highest levels of total flavonoids (54.5 mg/gdw), being a chrysin derivative the most abundant flavone found (22.3 mg/gdw). C. monogyna revealed the highest concentration in phenolic acids (5.5 mg/gdw) that were not found in C. multiflorus sample; 5-O-caffeoylquinic acid was the most abundant phenolic acid found in the first species, being a procyanidin trimer also found (1.4 mg/gdw). Kaempferol-3-O-rutinoside (0.84 mg/gdw) and quercetin-3-O-rutinoside (14.9 mg/gdw) were the main flavonols present in M. sylvestris and S. nigra, respectively. Due to the well established antioxidant activity of phenolic compounds, the studied wild medicinal flowers could be selected for processing extracts with health-promoting properties or to be incorporate into functional beverages or products with bioactive properties related to oxidative stress.

Characterisation of phenolic compounds in wild fruits from Northeastern Portugal

This study aimed to analyse the phenolic composition of wild fruits of Arbutus unedo (strawberry-tree), Prunus spinosa (blackthorn), Rosa canina and Rosa micrantha (wild roses). Analyses were performed by HPLC-DAD-ESI/MS. P spinosa fruits presented the highest concentration in phenolic acids (29.78 mg/100 g dry weight), being 3-O-caffeoylquinic acid the most abundant one, and flavone/ols (57.48 mg/100 g), among which quercetin3-O-rutinoside (15.63 mg/100 g) was the majority compound. (+)-Catechin was the most abundant compound in A. unedo (13.51 mg/100 g) and R. canina (3.59 mg/100 g) fruits. A. unedo fruits presented the highest concentration in flavan-3-ols (36.30 mg/100 g). Cyanidin 3-O-glucoside was found in all the studied fruits, being the major anthocyanin in most of them, with the exception of P. spinosa samples, in which cyaniding 3-O-rutinoside and peonidin 3-O-rutinoside predominated; P. spinosa fruit presented the more complex anthocyanin profile among the analysed fruits and also the highest anthocyanin concentrations, which was coherent with its greater pigmentation. All in all, P. spinosa presented the highest levels of phenolic acids and flavonoids, including anthocyanins, flavonols and flavones, although no flavan-3-ols could be identified in its fruits. The present study represents a contribution to the chemical characterisation of phenolic compounds from wild fruits with acknowledged antioxidant activity and traditionally used for several folk medicinal applications.