Alleviation of cadmium toxicity in pea (Pisum sativum L.) through Zn-Lys supplementation and its effects on growth and antioxidant defense.

Cadmium significantly impacts plant growth and productivity by disrupting physiological, biochemical, and oxidative defenses, leading to severe damage. The application of Zn-Lys improves plant growth while reducing the stress caused by heavy metals on plants. By focusing on cadmium stress and potential of Zn-Lys on pea, we conducted a pot-based study, organized under completely randomized block design CRD-factorial at the Botanical Garden of Government College University, Faisalabad. Both pea cultivars were grown in several concentrations of cadmium @ 0, 50 and 100 µM, and Zn-Lys were exogenously applied @ 0 mg/L and 10 mg/L with three replicates for each treatment. Cd-toxicity potentially reduces plant growth, chlorophyll contents, osmoprotectants, and anthocyanin content; however, an increase in MDA, H2O2 initiation, enzymatic antioxidant activities as well as phenolic, flavonoid, proline was observed. Remarkably, exogenously applied Zn-Lys significantly enhanced the plant growth, biomass, photosynthetic attributes, osmoprotectants, and anthocyanin contents, while further increase in enzymatic antioxidant activities, total phenolic, flavonoid, and proline contents were noticed. However, application of Zn-Lys instigated a remarkable decrease in levels of MDA and H2O2. It can be suggested with recommendation to check the potential of Zn-Lys on plants under cadmium-based toxic soil.

Phytochemical Analysis and Antioxidant Activity of Endophytic Fungi Isolated from Dillenia indica Linn.

Endophytic fungi live symbiotically inside plants and are hidden source of natural bioactive molecules. The present study was carried out to investigate the phytochemical analysis and antioxidant activity of endophytic fungi isolated from the ethnomedicinal plant Dillenia indica L. The ethyl acetate crude extracts of the endophytic fungal strains were preliminarily evaluated for their phytochemical analysis, and the results showed the presence of alkaloids, flavonoids, phenolics, terpene, and saponins. The crude extracts of more than 60% of the isolates showed 50-90% antioxidant activity by DPPH and H2O2 assay. The inhibition percentage of ethyl acetate extracts ranges from 34.05 to 91.5%, whereas IC50 values vary from 72.2 to 691.14%. Among all the strains, Fomitopsis meliae crude extract showed a maximum inhibition percentage, i.e., 91.5%, with an IC50 value of 88.27 µg/mL. Chaetomium globosum showed significant activity having an inhibition percentage of 89.88% and an IC50 value of 74.44 µg/mL. The total phenolic and flavonoid content in the crude extract of Chaetomium globosum was 37.4 mg gallic acid equivalent (GAE)/g DW and 31.0 mg quercetin equivalent (GAE)/g DW. GC-MS analysis of crude extract of C. globosum revealed different compounds, such as squalene; butanoic acid, 2-methyl-; hexadecanoic acid; 2-propanone, 1-phenyl-; 5-oxo-pyrrolidine-2-carboxylic acid methyl ester; 9,12-octadecadienoic acid (z)- etc. Many of these belong to phenolics, which are natural antioxidant compounds. The findings suggested that endophytic fungi associated with Dillenia indica L. can be a potential source of novel antioxidant compounds.

Effects of Exogenous Application of Glycine Betaine Treatment on 'Huangguoggan' Fruit during Postharvest Storage.

Loss of quality in citrus fruit is a common occurrence during postharvest storage due to oxidative stress and energy consumption. In recent years, glycine betaine (GB) has been widely applied to postharvest horticulture fruit. This study aimed to investigate the effect of GB treatment (10 mM and 20 mM) on the quality and antioxidant activity of 'Huangguogan' fruit during postharvest storage at room temperature. Our results indicated that both 10 mM and 20 mM treatments effectively reduced weight and firmness losses and maintained total soluble solid (TSS), titratable acidity (TA), and ascorbic acid contents. Additionally, GB treatment significantly increased the activity of antioxidant enzymes, maintained higher levels of total phenols and total flavonoids, and led to slower accumulation of H2O2. A transcriptome analysis conducted at 28 days after treatment (DAT)identified 391 differentially expressed genes (DEGs) between 20 mM GB (GB-2) and the control (CK) group. These DEGs were enriched in various pathways, particularly related to oxygen oxidoreductase, peroxidase activity, and flavonoid biosynthesis. Overall, the application of GB proved beneficial in enhancing the storability and extending the shelf life of 'Huangguogan' fruit.

Selenium nanoparticles based on Amphipterygium glaucum extract with antibacterial, antioxidant, and plant biostimulant properties.

BACKGROUND: In recent years, crop production has expanded due to the variety of commercially available species. This increase in production has led to global competition and the search for biostimulant products that improve crop quality and yield. At the same time, agricultural products that protect against diseases caused by phytopathogenic microorganisms are needed. Thus, the green synthesis of selenium nanoparticles (SeNPs) is a proposal for achieving these needs. In this research, SeNPs were synthesized from methanolic extract of Amphipterygium glaucum leaves, and chemically and biologically characterized. RESULTS: The characterization of SeNPs was conducted by ultraviolet-visible spectrophotometry (UV-Vis), scanning electron microscopy (SEM), electron microscopy transmission (TEM), Dynamic Light Scattering (DLS), energy dispersion X-ray spectroscopy (EDX), and infrared spectrophotometry (FTIR) techniques. SeNPs with an average size of 40-60 nm and spherical and needle-shaped morphologies were obtained. The antibacterial activity of SeNPs against Serratia marcescens, Enterobacter cloacae, and Alcaligenes faecalis was evaluated. The results indicate that the methanolic extracts of A. glaucum and SeNPs presented a high antioxidant activity. The biostimulant effect of SeNPs (10, 20, 50, and 100 µM) was evaluated in vinca (Catharanthus roseus), and calendula (Calendula officinalis) plants under greenhouse conditions, and they improved growth parameters such as the height, the fresh and dry weight of roots, stems, and leaves; and the number of flowers of vinca and calendula. CONCLUSIONS: The antibacterial, antioxidant, and biostimulant properties of SeNPs synthesized from A. glaucum extract demonstrated in this study support their use as a promising tool in crop production.

The Green Era of Food Packaging: General Considerations and New Trends.

Recently, academic research and industries have gained awareness about the economic, environmental, and social impacts of conventional plastic packaging and its disposal. This consciousness has oriented efforts towards more sustainable materials such as biopolymers, paving the way for the "green era" of food packaging. This review provides a schematic overview about polymers and blends of them, which are emerging as promising alternatives to conventional plastics. Focus was dedicated to biopolymers from renewable sources and their applications to produce sustainable, active packaging with antimicrobial and antioxidant properties. In particular, the incorporation of plant extracts, food-waste derivatives, and nano-sized materials to produce bio-based active packaging with enhanced technical performances was investigated. According to recent studies, bio-based active packaging enriched with natural-based compounds has the potential to replace petroleum-derived materials. Based on molecular composition, the natural compounds can diversely interact with the native structure of the packaging materials, modulating their barriers, optical and mechanical performances, and conferring them antioxidant and antimicrobial properties. Overall, the recent academic findings could lead to a breakthrough in the field of food packaging, opening the gates to a new generation of packaging solutions which will be sustainable, customised, and green.

Short-term responses of Spinach (Spinacia oleracea L.) to the individual and combinatorial effects of Nitrogen, Phosphorus and Potassium and silicon in the soil contaminated by boron.

While of lesser prevalence than boron (B) deficient soils, B-rich soils are important to study as they can cause B toxicity in the field and subsequently decrease crop yields in different regions of the world. We have conducted the present study to examine the role of the individual or combined application of silicon (Si) and NPK fertilizer in B-stressed spinach plants (Spinacia oleracea L.). S. oleracea seedlings were subjected to different NPK fertilizers, namely, low NPK (30 kg ha-2) and normal NPK (60 kg ha-2)], which were also supplemented by Si (3 mmol L-1), for varying levels of B in the soil i.e., 0, 250, and 500 mg kg-1. Our results illustrated that the increasing levels of B in the soil caused a substantial decrease in the plant height, number of leaves, number of stems, leaf area, plant fresh weight, plant dry weight, chlorophyll a, chlorophyll b, total chlorophyll, carotenoid content, net photosynthesis, stomatal conductance, transpiration rate, magnesium content in the roots, magnesium contents in the shoots, phosphorus content in the roots, phosphorus content in the leaves in the shoots, iron content in the roots, iron content in the shoots, calcium content in the roots, and calcium content in the shoots. However, B toxicity in the soil increased the concentration of malondialdehyde, hydrogen peroxide, and electrolyte leakage which were also manifested by the increasing activities of enzymatic [superoxidase dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX)], and non-enzymatic antioxidants (phenolic, flavonoid, ascorbic acid, and anthocyanin content). B toxicity in the soil further increased the concentration of organic acids in the roots such as oxalic acid, malic acid, formic acid, citric acid, acetic acid, and fumaric acid. The addition of Si and fertilizer levels in the soil significantly alleviated B toxicity effects on S. oleracea by improving photosynthetic capacity and ultimately plant growth. The increased activity of antioxidant enzymes in Si and NPK-treated plants seems to play a role in capturing stress-induced reactive oxygen species, as was evident from the lower levels of oxidative stress indicators, organic acid exudation, and B concentration in the roots and shoots of Si and NPK-treated plants. Research findings, therefore, suggested that the Si and NPK application can ameliorate B toxicity in S. oleracea seedlings and result in improved plant growth and composition under metal stress as depicted by the balanced exudation of organic acids.

Effect of different seed priming agents on chromium accumulation, oxidative defense, glyoxalase system and mineral nutrition in canola (Brassica napus L.) cultivars.

The present experiment was conducted to appraise the role of different seed priming agents in circumventing the negative impact of chromium (Cr) toxicity on canola plants. Chromium toxicity resulted in significant decline in photosynthetic pigments and growth attributes of two canola cultivars (Puriga and MS-007). Cr toxicity also resulted in higher oxidative stress mirrored as greater accumulation of hydrogen peroxide (H2O2) superoxide radical (O2•‒), electrolyte leakage (EL) and malondialdehyde (MDA). Further, lipoxygenase enzyme activity that catalyzes the peroxidation of membrane lipids was also enhanced due to Cr toxicity. Canola plants also manifested impaired methylglyoxal (MG) detoxification due to the downregulation of glyoxalase enzymes (GlyI and II) under Cr stress. Seed priming treatments viz. osmo-priming with calcium chloride (CaCl2) and hormonal priming with salicylic acid (SA) remarkably improved growth and chlorophyll content in both canola cultivars under Cr toxicity as compared to other priming treatments such as hydro-priming, redox priming (H2O2) and chemical priming (Se; selenium). Moreover, CaCl2 and SA seed priming also resulted in lower oxidative stress and improved enzymatic (SOD, POD, CAT, APX, GR, GST) and non-enzymatic (GSH, phenolics, flavonoids, proline) antioxidant system of both cultivars under Cr toxicity. Further, hormonal and osmo-priming strengthened glyoxalase and antioxidant systems, thus improving reactive oxygen species (ROS) and MG detoxification. In this background, the cultivar Puriga is considered Cr tolerant as it exhibited better growth and lesser oxidative stress in both seed priming and non-primed conditions under Cr toxicity than cv. MS-007.

Potential Properties of Natural Nutraceuticals and Antioxidants in Age-Related Eye Disorders.

Eye health is crucial, and the onset of diseases can reduce vision and affect the quality of life of patients. The main causes of progressive and irreversible vision loss include various pathologies, such as cataracts, ocular atrophy, corneal opacity, age-related macular degeneration, uncorrected refractive error, posterior capsular opacification, uveitis, glaucoma, diabetic retinopathy, retinal detachment, undetermined disease and other disorders involving oxidative stress and inflammation. The eyes are constantly exposed to the external environment and, for this reason, must be protected from damage from the outside. Many drugs, including cortisonics and antinflammatory drugs have widely been used to counteract eye disorders. However, recent advances have been obtained via supplementation with natural antioxidants and nutraceuticals for patients. In particular, evidence has accumulated that polyphenols (mostly deriving from Citrus Bergamia) represent a reliable source of antioxidants able to counteract oxidative stress accompanying early stages of eye diseases. Luteolin in particular has been found to protect photoreceptors, thereby improving vision in many disease states. Moreover, a consistent anti-inflammatory response was found to occur when curcumin is used alone or in combination with other nutraceuticals. Additionally, Coenzyme Q10 has been demonstrated to produce a consistent effect in reducing ocular pressure, thereby leading to protection in patients undergoing glaucoma. Finally, both grape seed extract, rich in anthocyanosides, and polynsatured fatty acids seem to contribute to the prevention of retinal disorders. Thus, a combination of nutraceuticals and antioxidants may represent the right solution for a multi-action activity in eye protection, in association with current drug therapies, and this will be of potential interest in early stages of eye disorders.

In vitro Bioaccessibility of Proteins, Phenolics, Flavonoids and Antioxidant Activity of Amaranthus viridis.

The aim of this study was to investigate the chemical composition, mineral content and report the effect of gastrointestinal digestion on the proteins, phenolics, flavonoids and antioxidant activity of morphological parts of Amaranthus viridis (AV). The macronutrients found in the greatest quantity were proteins (leave, inflorescence and seeds) and fiber (roots and stem). The main minerals were calcium, potassium, iron and zinc. All the plant parts showed total phenolic (TPC) and total flavonoid (TFC) compounds that were correlated with antioxidant capacity (DPPH, ABTS and FRAP); the leaves and inflorescence presented the greatest potential. The antioxidant compounds from the leaves, inflorescence and roots decreased after static in vitro digestion, while the TPC and TFC of the digested seeds increased by more than 55%. Approximately 90% of the protein content was bioaccessible. To the best of our knowledge, for the first time, the bioaccessibility of proteins, phenolics, flavonoids and antioxidant activity of the leave, inflorescence, seeds, roots and stem from AV were reported. Taking into consideration the excellent nutritional properties, the morphological parts of the plant can be potentially explored as a source of protein, fiber, minerals and antioxidant compounds.

Preliminary Study on Pasta Samples Characterized in Antioxidant Compounds and Their Biological Activity on Kidney Cells.

Pasta is one of the basic foods of the Mediterranean diet and for this reason it was chosen for this study to evaluate its antioxidant properties. Three types of pasta were selected: buckwheat, rye and egg pasta. Qualitative-quantitative characterization analyses were carried out by HPLC-DAD to identify antioxidant compounds. The data showed the presence of carotenoids such as lutein and polyphenols such as indoleacetic acid, (carotenoids from 0.08 to 0.16 mg/100 g, polyphenols from 3.7 to 7.4 mg/100 g). To assess the effect of the detected metabolites, in vitro experimentation was carried out on kidney cells models: HEK-293 and MDCK. Standards of ß-carotene, indoleacetic acid and caffeic acid, hydroalcoholic and carotenoid-enriched extracts from samples of pasta were tested in presence of antioxidant agent to determine viability variations. ß-carotene and indoleacetic acid standards exerted a protective effect on HEK-293 cells while no effect was detected on MDCK. The concentrations tested are likely in the range of those reached in body after the consumption of a standard pasta meal. Carotenoid-enriched extracts and hydroalcoholic extracts showed different effects, observing rescues for rye pasta hydroalcoholic extract and buckwheat pasta carotenoid-enriched extract, while egg pasta showed milder dose depending effects assuming pro-oxidant behavior at high concentrations. The preliminary results suggest behaviors to be traced back to the whole phytocomplexes respect to single molecules and need further investigations.

Comparative assessment of nutritional composition, polyphenol profile, antidiabetic and antioxidative properties of selected edible wild plant species of Bangladesh.

Wild edible plants are recently recognized as an important source of acquiring macro and micro nutrients beneficial for human health. Hence, the present study was undertaken to assess the antidiabetic and antioxidant potentials, polyphenolic profile, - as well as the ascorbic acid, proximate and mineral compositions of five selected Bangladeshi wild plants. The studied samples were rich in ash, fiber, protein, vitamin C and low in fat. The undertaken plant samples were found to have good amounts of total phenolic, total flavonoid, and antioxidant capacities, documented by DPPH, FRAP, and TEAC assays. They also exhibited varying spectrum of polyphenols estimated by HPLC. Significant inhibition of α-amylase activity by plant extracts was also observed. Evaluation by principal component analysis revealed clear separation among the wild plant varieties. The study findings would enrich the food composition table of Bangladesh and allow the population to consume more wild plants and increase their production.

Evaluation of the Behavior of Phenolic Compounds and Steviol Glycosides of Sonicated Strawberry Juice Sweetened with Stevia (Stevia rebaudiana Bertoni).

In this study, the influence of stevia addition and sonication processing parameters on the phenolic content and profile as well as the steviol glycosides of strawberry juice-based samples was investigated. For this purpose, three matrices-control samples of strawberry juices without green stevia powder (CS), strawberry juices with green stevia powder (JGSP), and sonicated juices with green stevia powder (SJGSP)-were prepared. For sonication purposes, different conditions regarding probe diameters (7 mm and 22 mm), amplitudes (50%, 75%, and 100%), and time (15 min, 20 min, and 25 min) were tested. The results that were obtained upon the measurement of the total phenolic content, total flavonoids, steviol glycosides, and antioxidant capacity showed significant differences according to the matrices evaluated, obtaining overall higher values in the samples with stevia added. Moreover, when sonication was evaluated, it was found that a higher amplitude (100%), a larger probe diameter (22 mm), and a longer sonication period (25 min) led to higher values. Flavones such as luteolin and apigenin were identified and quantified in JGSP and SJGSP, while they were not found in CS. Besides these phenolic compounds, kaempferol, quercetin, pyrogallic acid, 4-methylcatechol, and 4-methoxybenzoic acid were also identified and quantified. Similarly to the total phenolic compounds, total flavonoids, and total antioxidant capacity, an increased amount of these compounds was found in SJGSP, especially after using the most intense sonication conditions. Therefore, the use of sonication together with stevia added could be a useful tool to preserve strawberry juices, increasing at the same time the sweetness and the antioxidant value of the beverages.

Grape seed pomace as a valuable source of antioxidant fibers.

BACKGROUND: Grape seed pomace is a valuable by-product from winery/oil extraction industries and a potential source of bioactive compounds. Hot water extraction is the most widely used technology for polysaccharide recovery from plant and seed sources. To obtain an antioxidant fiber-rich extract (AFE) from defatted grape seed pomace, seed:water ratio (1:10-1:20) and extraction temperature (70-90 °C) effects on extraction yield, total sugars, total phenolic compounds and condensed tannins were investigated. RESULTS: The best extraction results were achieved at the highest seed:water ratio and temperature. At these conditions, the extraction yield and total sugar content of the freeze-dried AFE were 100.1 g kg-1 and 725.3 g kg-1 , respectively. The AFE presented high total phenolic content (16.2 g GAE kg-1 ), condensed tannins (515.1 mg CAT kg-1 ), soluble (219.1 g kg-1 ) and insoluble fibers (132.4 g kg-1 ), besides protein (171.1 g kg-1 ) and ash (152.6 g kg-1 ). The antioxidant capacity of the AFE was 382.7, 823.7 and 1439.4 µmol Trolox g-1 for DPPH, ABTS and ORAC methods. CONCLUSION: The AFE was shown to be not only an interesting source of fiber with antioxidant capacity, which could become a potential foodstuff, but also an ecofriendly and sustainable alternative to use grape seed pomace. © 2019 Society of Chemical Industry.

Can nutrient fertilization mitigate the effects of ozone exposure on an ozone-sensitive poplar clone?

We tested the independent and interactive effects of nitrogen (N; 0 and 80 kg ha-1), phosphorus (P; 0, 40 and 80 kg ha-1), and ozone (O3) application/exposure [ambient concentration (AA), 1.5 × AA and 2.0 × AA] for five consecutive months on biochemical traits of the O3-sensitive Oxford poplar clone. Plants exposed to O3 showed visible injury and an alteration of membrane integrity, as confirmed by the malondialdehyde by-product accumulation (+3 and +17% under 1.5 × AA and 2.0 × AA conditions, in comparison to AA). This was probably due to O3-induced oxidative damage, as documented by the production of superoxide anion radical (O2-, +27 and +63%, respectively). Ozone per se, independently from the concentrations, induced multiple signals (e.g., alteration of cellular redox state, increase of abscisic acid/indole-3-acetic acid ratio and reduction of proline content) that might be part of premature leaf senescence processes. By contrast, nutrient fertilization (both N and P) reduced reactive oxygen species accumulation (as confirmed by the decreased O2- and hydrogen peroxide content), resulting in enhanced membrane stability. This was probably due to the simultaneous involvement of antioxidant compounds (e.g., carotenoids, ascorbate and glutathione) and osmoprotectants (e.g., proline) that regulate the detoxification processes of coping with oxidative stress by reducing the O3 sensitivity of Oxford clone. These mitigation effects were effective only under AA and 1.5 × AA conditions. Nitrogen and P supply activated a free radical scavenging system that was not able to delay leaf senescence and mitigate the adverse effects of a general peroxidation due to the highest O3 concentrations.

Comparing the Effects of Chlorogenic Acid and Ilex paraguariensis Extracts on Different Markers of Brain Alterations in Rats Subjected to Chronic Restraint Stress.

Positive influence of yerba mate (Ilex paraguariensis) on human health issues has been attributed to its frequent consumption in South American countries and is assumed to be due to its high content of antioxidant compounds, including chlorogenic acid (CGA); however, hard evidence about its positive effects under chronic stress conditions is still required. In this study, the effects of yerba mate extracts (IpE), and its main compound chlorogenic acid (CGA), on behavioral and morphological endpoints of brain damage induced by chronic restraint stress (CRS) to rats were evaluated and compared. CRS sessions were performed during 21 days. IpE (200 mg/mL, p.o.) or CGA (2 mg/mL, p.o.) were administered daily 30 min before stress. Behavioral tests comprised motor skills and anxiety-like activity. Histological (H&E) and histochemical changes were explored in three brain regions: cortex (Cx), hippocampus (Hp), and striatum (S). Rats subjected to CRS exhibited hypoactive patterns of locomotor activity. Rats receiving IpE before CRS preserved the basal locomotor activity. Stressed animals also augmented the anxiety-like activity, whereas IpE normalized exploratory behavior. Stressed animals presented cell damage in all regions. Morphological damage was more effectively prevented by IpE than CGA. Stressed animals also augmented the expression/localization pattern of the tumor necrosis factor alpha in the striatum and the expression of the glial fibrillary acidic protein in the hippocampus (stratum moleculare) and cortex, whereas IpE and CGA reduced the expression of these molecules. In turn, CGA exhibited only moderate protective effects on all markers analyzed. Our findings support a protective role of IpE against CRS, which may be related to the antioxidant and anti-inflammatory properties of its compounds. Since CGA was unable to prevent all the alterations induced by CRS, it is concluded that the protective properties of the whole extract of Ilex paraguariensis are the result of the combined effects of all its natural antioxidant compounds, and not only of the properties of CGA.

The Selenium Supplementation Influences Olive Tree Production and Oil Stability Against Oxidation and Can Alleviate the Water Deficiency Effects.

Foliar fertilization with selenium (Se) may well be beneficial in increasing the nutritional and qualitative values of food in Se-deficient regions such as the Mediterranean Basin, and may contribute to an increase in drought resistance in plants. The present study has considered detachment force, flesh firmness, pigmentation, fresh and dry weight, and oil content of olive drupes from Se fertilized olive orchards (Olea europaea L.) under drought stress and well-watered conditions. This study has also evaluated the total Se, Se amino acid, phenol, carotenoid and chlorophyll contents of EVOO, plus its oxidative stability against oxidation. While there was no change in the ripening indexes and the production of olives generally, Se application did increase the total Se, Se methionine, phenol, and carotenoid and chlorophyll contents. The higher concentration of these (bio) chemical compounds in EVOO obtained from Se fertilized plants might well suggest enhanced antioxidant activity. Consequently, EVOO obtained from Se fertilized trees possesses a higher nutritional value and, as indicated by the greater oxidative stability against oxidation, longer shelf life. In addition, under water deficient conditions, a higher fresh olive weight corresponds to a higher level of phenol, carotenoid and chlorophyll, and the chlorophyll-to-carotenoid ratio in Se fertilized trees would appear to confirm the positive role of selenium in alleviating damage caused by drought stress conditions.

Preparative isolation of antioxidative compounds from Dracocephalum heterophyllum using off-line two-dimensional reversed-phase liquid chromatography/hydrophilic interaction chromatography guided by on-line HPLC-DPPH assay.

Traditional Tibetan medicine (TTM) has been valuable for the identification of new therapeutic leads. Nevertheless, reports about the chemical constituents of TTM are meager owing to the lack of suitable purification techniques. In this study, an off-line two-dimensional reversed-phase/hydrophilic interaction liquid chromatography (2D RP/HILIC) technique guided by on-line HPLC-DPPH has been established for the isolation of pure antioxidants from the extract of Dracocephalum heterophyllum. According to the chromatographic recognition outcome of the HPLC-DPPH system, the first-dimensional (1D) separation on the Megress C18 preparative column yielded 6 antioxidative fractions (61.4% recovery) from the ethyl acetate fraction (6.1 g). In the second-dimensional (2D) separation, a HILIC XAmide preparative column was employed. In total, 8 antioxidants were isolated from D. heterophyllum with a purity of >95%, which indicated the efficiency of the developed method to prepare antioxidative compounds with high purity from plant extracts. In addition, this method was highly efficient for the preparation of structural analogues of the antioxidative polyphenols and could be applied for the purification of structural analogues from other resources.

Exploring New Antioxidant and Mineral Compounds from Nymphaea alba Wild-Grown in Danube Delta Biosphere.

Nymphaea alba is an aquatic flowering plant from the Nymphaeaceae family that has been used for hundreds of years in traditional herbal medicine. The plant is characterized by different phytochemicals, depending on the geographical location. Herein, we have carried out, for the first time, the separation and HPLC-MS/MS identification of some antioxidant compounds, such as polyphenols and flavonoids from N. alba extracts from the Danube Delta Biosphere, and investigated their possible antiradical properties. An ultrasonic method has been exhaustively used for the extraction of the antioxidant compounds from the different anatomic parts of N. alba (fruit, flower, leaf, stem, and root). The extracts that were obtained using ultrasound irradiation showed a large polyphenol (19.42 mg EqGA/100 mg extract) and flavonoid (0.97 mg EqQ/100 mg extract) content. The fruit and flower extracts showed the highest antioxidant activity index (AAI). Among the 27 phytochemical compounds identified in all of the N. alba extracts, rutin and p-coumaric acid were found as the major components. The content of macroelements and microelements in N. alba extracts were compared, and it was found that their concentrations depend on the different anatomic parts of the plant. This research contributes to the study of Nymphaeaceae family, being the first exhaustive phytochemical study of N. alba from a wild population in Romania.

An Insight into Sargassum muticum Cytoprotective Mechanisms against Oxidative Stress on a Human Cell In Vitro Model.

Sargassum muticum is a brown seaweed with strong potential to be used as a functional food ingredient, mainly due to its antioxidant properties. It is widely used in traditional oriental medicine for the treatment of numerous diseases. Nevertheless, few studies have been conducted to add scientific evidence on its effects as well as on the mechanisms of action involved. In this work, the human cell line MCF-7 was used as an in vitro cellular model to evaluate the capability of Sargassum muticum enriched fractions to protect cells on an oxidative stress condition. The concentration of the bioactive compounds was obtained by vacuum liquid chromatography applied on methanol (M) and 1:1 methanol:dichloromethane (MD) crude extracts, resulting in seven enriched fractions from the M extraction (MF2-MF8), and eight fractions from the MD extraction (MDF1-MDF8). All fractions were tested for cytotoxic properties on MCF-7 cells and the nontoxic ones were tested for their capacity to blunt the damaging effects of hydrogen peroxide-induced oxidative stress. The nontoxic effects were also confirmed in 3T3 fibroblast cells as a nontumor cell line. The antioxidant potential of each fraction, as well as changes in the cell's real-time hydrogen peroxide production, in the mitochondrial membrane potential, and in Caspase-9 activity were evaluated. The results suggest that the protective effects evidenced by S. muticum can be related with the inhibition of hydrogen peroxide production and the inhibition of Caspase-9 activity.

Discarded seeds from red pepper (Capsicum annum) processing industry as a sustainable source of high added-value compounds and edible oil.

The chemical composition and the antioxidant properties of Capsicum annum discarded seeds from processing industry with their corresponding extracted oil were investigated. C. annum seeds had high levels of crude proteins (18.30%), crude oil (11.04%), and dietary fibers (60.96%). The lipophilic fraction of C. annum seeds showed higher radical scavenging activity compared to their hydrophilic fraction, while this latter exhibited the highest reducing power. The results of fatty acid composition showed that fatty acids present in C. annum seed oil were mainly polyunsaturated (84.23%), with linoleic acid being the major polyunsaturated fatty acid (70.93%). The major monounsaturated fatty acid was oleic acid (12.18%), while the main saturated fatty acid was palmitic acid (11.90%). C. annum seed oil showed high absorbance in the UV-B, UV-A, and visible ranges. Owing to their composition, C. annum seeds discarded from pepper processing industry as by-product could be potentially used as high added-value ingredients in some food or nutraceutical formulations because they are well endowed with essential nutriments required for human health.