Use of inert gas-enriched atmospheres and post-packaging irradiation in shelf-life extension of watercress

Watercress (Nasturtium officinale R. Br.) is a semi-aquatic plant of the Brassicaceae family highly appreciated in the Mediterranean cuisine. It features sharp, peppery and slightly tangy taste and contains health-promoting phytochemicals. Its consumption as a fresh-cut product has increased in recent years, as well as the global market of minimally processed vegetables. This demand is driven by the growing interest in the role of food in promoting the human health and wellbeing and to meet consumer needs for fresh-like and more convenient foods. Due to the reduced shelf-life of this plant, the suitability of inert gas-enriched atmospheres and ionizing irradiation for preserving visual, nutritional and functional quality attributes during cold storage was studied. Watercress samples were gathered in the Northeast region of Portugal, rinsed in tap water and a portion was immediately analyzed (non-stored control). The remaining fresh material was packaged in polyethylene bags under N2- and Ar-enriched atmospheres, conventional atmosphere (air) and vacuum (no atmosphere). Samples under conventional atmosphere were irradiated at 1, 2 and 5 kGy of gamma-rays (predicted doses) in a 60Co experimental chamber. A non-irradiated control followed all the experiment. Then, all packaged samples were stored at 4 ºC for 7 days. The studied quality parameters included the colour that was measured with a Konica Minolta colorimeter, and total soluble solids and pH determined in squeezed juice. The proximate composition (moisture, proteins, fat, ash, carbohydrates and energy) was evaluated using the AOA C procedures. Organic acids, free sugars, fatty acids and tocopherols were analyzed by chromatographic techniques. Samples were also evaluated for its DPPH• scavenging activity, reducing power, and lipid peroxidation inhibition capacity trough the inhibition of the β-carotene bleaching and thiobarbituric acid reactive substances (TBAR S) formation. Differences among treatments were analyzed using the one-way analysis of variance (ANO VA) and a linear discriminant analysis (LDA ) was used to evaluate the effects on the overall postharvest quality. After evaluating the effect on the individual quality parameters, the LDA revealed that the Ar-enriched atmosphere and the irradiation at 2 kGy were suitable processing choices for preserving the integrity of the non-stored control samples during cold storage. Thus, these non-thermal treatments were highlighted for shelf-life extension of fresh-cut watercress.

Chemical composition of Boletus pinophilus and Clitocybe subconnexa: preservation with gamma irradiation

The short shelf life of mushrooms is a barrier for their distribution and, therefore, there has been extensive research to find technologies that ensure the preservation of mushrooms, maintaining their organoleptic and nutritional properties (1]. Irradiation has proved its technological feasibility to be safely used in the reduction of food losses, being recognized by international organizations as a valid conservation alternative in extending shelflife of many foods. The aim of the present work was to validate the use of 2 kGy dose of gamma radiation to maintain chemical composition of wild mushrooms. Boletus pinophilus Pihit & Dermek and Clitocybe subconnexa Murrill wild samples were obtained in Tnis-os-Montes; subsequently, the samples were divided in two groups: control (non-irradiated, 0 kGy) and irradiated (2 kGy). The irradiation of the samples was performed in a 6°Co experimental chamber. Moisture, protein, fat, carbohydrates and ash were determined following the standard procedures [2]. Free sugars and tocopherols were determined by high performance liquid chromatography coupled to a refraction index detector (HPLC-RI) and a fluorescence detector, respectively; fatty acids were determined by gas-liquid chromatography with flame ionization detection (GC-FID) [3]. The protein and ash content was preserved in both groups, although the sugars and tocopherols decreased in the irradiated samples. Sugars and fatty acids showed significant changes after irradiation treatment, particularly in B. pinophillus, nevertheless, the magnitude of the obtained differences did not seem to be sufficient to affect the chemical profiles of the assayed mushrooms. Overall, the detected chemical changes might be considered as allowable, in view of the high advantages offered by gamma irradiation at decontamination and/or disinfestation level.

Effects of electron-beam irradiation on chemical and antioxidation parameters of wild Macrolepiota procera dried samples

Mushrooms are very perishable foods due to their high susceptibility to moisture loss, changes in color and texture, or microbiological spoilage. Drying is considered as the most appropriate method to prevent these alterations, but it has some limitations, such as shrinkage, enzymatic and non-enzymatic browning reactions, and oxidation of lipids and vitamins. Irradiation might effectively attenuate the undesirable changes caused by drying process, ensuring also higher shelf-life of mushrooms and their decontamination [I]. In the present work, the combined effects of electron-beam irradiation (at 0, 0.5, 1 and 6 kGy doses) and storage time (at 0, 6 and 12 months) were evaluated and compared. Macrolepiota procera (Scop.) Singer wild samples were obtained in Tnis-os-Montes, in the Northeast of Portugal, and dried at 30 •c in an oven. Subsequently, the samples were divided in four groups: control (non-irradiated, 0 kGy); sample 1 (0.5 kGy); sample 2 (1 kGy) and sample 3 (6 kGy). The irradiation was performed at the lNCTInstitute of Nuclear Chemistry and Technology (lNCT), in Warsaw, Poland. Moisture, protein, fat, carbohydrates and ash were determined following standard procedures. Free sugars and tocopherols were determined by high performance liquid chromatography coupled to a refraction index detector (HPLC-RI) and a fluorescence detector, respectively; fatty acids were determined by gas-liquid chromatography with flame ionization detection (GC-FID). Antioxidant activity was evaluated in the methanolic extracts by in vitro assays measuring DPPH (1,1-diphenyl-2- picrylhydrazyl) radical scavenging activity, reducing power, inhibition of ~-carotene bleaching and inhibition oflipid peroxidation using thiobarbituric acid reactive substances (TBARS) assay. Total phenolics were also determined by the Folin-Ciocalteu assay. All the parameters showed a decrease tendency with storage time. Trehalose and y-tocopherol were preserved with 1 kGy dose. Electron-beam irradiation did not impart additional changes to most of the chemical and antioxidant parameters of M. procera dried samples. This is a very promising result, since electron-beam irradiation might attenuate most unwanted changes caused by drying, maintaining its long-term effectiveness.

Chromatographic determination of fatty acids profiles in regionally-produced Portuguese goat cheese along curing

The biochemistry of cheese ripening involves mechanisms such as glycolysis, proteolysis and lipolysis. Fatty acids are released by the action of lipases from different sources, milk, rennet, bacteria, moulds included as secondary starters, and other exogenous lipases, during lipolysis [1]. The composition of the lipid fraction contributes positively to the flavour of cheese, for being precursors of more complex aroma compounds responsible for the characteristic “goaty flavour” of goat cheeses [2]. Goat milk is recognized by its easier digestibility, alkalinity, buffering capacity and certain therapeutic values in medicine and human nutrition [3]. A high total content of fatty acids is strongly linked to a rancid and tart off flavour in goat milk and may be considered undesirable in most cheese varieties [4]. In this sense, the purpose of the present study was to examine the composition and changes in fatty acids and saponification value of goat cheese during curing period (2, 7 and 12 months). Goat cheese was made in industrial unit of Cachão - Mirandela (Trás-os- Montes) with raw milk Serrana goats’ race, salt and rennet from animal origin. During the first two months, the samples were stored in a ripening chamber (9.5-11 °C and RH 75-85%). From the second month to one year, the samples were stored in a preservation chamber (10.5-12 °C and RH 75-85%). The fatty acids profile of the inner part of the cheese was analyzed by gas-chromatography coupled to flame ionization detection (GC-FID). The degree of saponification was determined both in the crust and inside the cheese by HCl titration of ethanol KOH solution of the samples. Twenty-six fatty acids (FA) were identified and quantified in the inner part of the cheese (total fat was 45-46 g/100 g during the curing period). Saturated fatty acids (SFA) did not change up to 7 months of curing, increasing only after 12 months, being palmitic (C16:0), stearic (C18:0), myristic (C14:0) and capric (C10:0) acids the most abundant FA in this class. Monounsaturated fatty acids (MUFA) decreased only after 12 months, and oleic acid (C18:1) was the predominant FA. In polyunsaturated fatty acids (PUFA) class, the most abundant were linoleic (C18:2) and linolenic (C18:3) acids, and followed the same tendency of MUFA. This is corroborated by an increase in the degree of saponification, either in the crust as in the inner part of the cheese, after 12 months of curing, probably related with the saturation of the fatty acids [3]. Extra-long curing can be done in cheeses produced with goat milk up to seven months of storage without changing the total fat and individual FA content.

Bioactivity and chromatographic analysis of nutrients and non-nutrients of two Leccinum species

Nowadays the rising cost of health care and pharmaceutical products, the increase in life expectancy as well as the demand for an improved quality of life, has led to an increased concern about food intake and an emergence of new concepts of nutrition [1]. Mushrooms have been pointed out as an excellent option to include in a healthy diet, due to their nutritional value [2] associated with their bioactive properties [3]. The current study presents the chemical profile of two edible species, Leccinum molle (Ban) Ban and Leccinum vulpinum Watling, harvested in the outskirts of Bragan9a (Northeastern Portugal), regarding their content in nutrients and nonnutrients. Individual profiles of sugars and fatty acids were obtained by HPLC-RI and GC-FID, respectively. Tocopherols were analysed by HPLC-fluorescence, and the non-nutrients (i.e., phenolic and other organic acids) by HPLC-PDA. The antioxidant activity of the methanolic extracts obtained from both species was assessed with different assays (e.g. reducing power, radical scavenging activity and lipid peroxidation inhibition) and their hepatotoxicity was evaluated in primary cell cultures obtained from porcine liver, PLP2. Generally, both Leccinum species revealed similar nutrient profiles, with low fat levels, fructose, mannitol and trehalose as the foremost free sugars, and higher percentages of mono- and polyunsaturated fatty acids in comparison with saturated fatty acids. The presence of bioactive compounds was also detected, namely phenolic (e.g., gallic, protocatechuic and p-hydroxybenzoic acids) and organic acids (e.g., citric and fumaric acids). Both species presented antioxidant properties, being L. vulpinum the species which showed the most promising results (higher contents of total phenolic acids and lower ECso values in all the performed assays). Neither of the extracts presented toxicity against the liver primary cells PLP2, up to maximal concentration tested (Giso > 400 μg/ml).

Postharvest changes in profiles of sugars, organic acids and tocopherols in leafy vegetables monitored by chromatographic techniques

After harvest, plants remain living organisms with the capacity to carry out metabolic processes. Thus, from the moment they are detached from the source of nutrients, they become entirely dependent on their own organic reserves [1]. Postharvest changes cannot be stopped, but they can be slowed within certain limits. Therefore, this study was conducted to evaluate the effects induced by storage in the profiles of sugars, organic acids and tocopherols of two leafy vegetables. Wild samples of watercress (Nasturtium officinale R. Br.) and buckler sorrel (Rumex induratus Boiss. & Reut.), from the Northeastern region of Portugal, were analyzed after harvest (control) and after storage in sterilized packages (using the passive modification mode) at 4ºC for 7 or 12 days, respectively. Analyses were performed by high-performance liquid chromatography (HPLC) using different detectors, i.e., a refraction index detector (RID) for free sugars, a photodiode array detector (PDA) for organic acids, and a fluorescence (FP) detector for tocopherols. The storage time decreased the levels of fructose, glucose and total sugars in both leafy vegetables and increased the total organic acids content. The decrease of these sugars can be related to its use by the plant to produce the required energy. Ascorbic acid was detected in buckler sorrel and decreased with storage; while the amount of malic acid increased in both species. Curiously, all the tocopherol isoforms increased in watercress, while buckler sorrel just present higher values of γ- and δ- tocopherols. In fact, the de novo synthesis of these bioactives compounds can be a plant strategy to fight against the reactive species that are produced during storage. The knowledge of the behavior of these compounds during storage that was achieved with this study [2] may contribute to the development of more effective preservation strategies for leafy vegetables.

Nutritional and biochemical profiling of Leucopaxillus candidus (Bres.) singer wild mushroom

The wild mushroom Leucopaxillus candidus (Bres.) Singer was studied for the first time to obtain information about its chemical composition, nutritional value and bioactivity. Free sugars, fatty acids, tocopherols, organic and phenolic acids were analysed by chromatographic techniques coupled to different detectors. L. candidus methanolic extract was tested regarding antioxidant potential (reducing power, radical scavenging activity and lipid peroxidation inhibition). L. candidus was shown to be an interesting species in terms of nutritional value, with high content in proteins and carbohydrates, but low fat levels, with the prevalence of polyunsaturated fatty acids. Mannitol was the most abundant free sugar and β-tocopherol was the main tocopherol isoform. Other compounds detected were oxalic and fumaric acids, p-hydroxybenzoic and cinnamic acids. The methanolic extract revealed antioxidant activity and did not show hepatoxicity in porcine liver primary cells. The present study provides new information about L. candidus.

Leccinum molle (Bon) Bon and Leccinum vulpinum Watling: The First Study of Their Nutritional and Antioxidant Potential

This work presents the chemical profile of two edible species of mushrooms from the genus Leccinum: Leccinum molle (Bon) Bon and Leccinum vulpinum Watling, both harvested on the outskirts of Bragança (Northeastern Portugal). Both species were prepared and characterized regarding their content in nutrients (i.e., free sugars, fatty acids and vitamins), non-nutrients (i.e., phenolic and other organic acids) and antioxidant activity. To the best of our knowledge, no previous studies on the chemical characterization and bioactivity of these species have been undertaken. Accordingly, this study intends to increase the available information concerning edible mushroom species, as well as to highlight another important factor regarding the conservation of the mycological resources--their potential as sources of nutraceutical/pharmaceutical compounds. Overall, both species revealed similar nutrient profiles, with low fat levels, fructose, mannitol and trehalose as the foremost free sugars, and high percentages of mono- and polyunsaturated fatty acids. They also revealed the presence of bioactive compounds, namely phenolic (e.g., gallic acid, protocatechuic acid and p-hydroxybenzoic acid) and organic acids (e.g., citric and fumaric acids) and presented antioxidant properties.