Variability in Biological Activities of Satureja montana Subsp. montana and Subsp. variegata Based on Different Extraction Methods.

Winter savory (Satureja montana L.) is a well-known spice and medicinal plant with a wide range of activities and applications. Two subspecies of S. montana, subsp. montana and subsp. variegata, were used for the preparation of seven different extracts: steam distillation (essential oil (EO) and hydrolate (HY)), subcritical water (SWE), ultrasound-assisted (UAE-MeOH and UAE-H2O), and microwave-assisted (MAE-MeOH and MAE-H2O) extraction. The obtained EOs, HYs, and extracts were used for an in vitro evaluation of the antioxidant activity (DPPH, ABTS, reducing power, and superoxide anion methods) and in vitro antimicrobial activity against Bacillus cereus, Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Salmonella Typhimurium, Saccharomyces cerevisiae, and Candida albicans. The antimicrobial screening was conducted using disk-diffusion assessment, minimal inhibitory concentration, time-kill kinetics modeling, and pharmacodynamic study of the biocide effect. The total phenolic content (TPC) was highest in EO, followed by SWE, MAE, and UAE, and the lowest was in HY. The highest antimicrobial activity shows EO and SWE for both varieties, while different UAE and MAE extracts have not exhibited antimicrobial activity. The natural antimicrobials in the S. montana extract samples obtained by green extraction methods, indicated the possibility of ecologically and economically better solutions for future in vivo application of the selected plant subspecies.

Composition, texture, sensorial quality, and biological activity after in vitro digestion of durum wheat pasta enriched with carrot waste extract encapsulates.

Aim of this research was to evaluate durum-wheat pasta enriched with encapsulated carrot waste extracts in oil, obtained by freeze drying (FDE) or spray drying techniques (SDE). Five pastas (control, enriched with 10% FDE, or 10% SDE, or 20% FDE, or 20% SDE) were evaluated for furosine, carotenoids, tocols, colour, in vitro bioactivities, cooking performance, texture and sensory quality. The encapsulates added to the enriched pastas α-carotene (0.58-1.24 mg/kg), ß-carotene (1.43-3.29 mg/kg), cis-ß-carotene (0.51-1.11 mg/kg) and total tocols (10.9-33.6 mg/kg). The carotenes were stable and the tocols diminished (-13%) during pasta manufacturing; both decreased (2-18% and 4-15%, respectively) during cooking, but they were still more abundant in the enriched pastas. Antioxidant, anti-hyperglycaemic, anti-inflammatory and anti-proliferative activities after in vitro digestion of cooked pastas improved, while sensory acceptability of control and 10% enriched pastas were similar. The encapsulates addition significantly improved the nutritional and technological qualities of durum-wheat pasta.

Encapsulation of beetroot juice: a study on the application of pumpkin oil cake protein as new carrier agent.

Aim: In the present study, beetroot juice was encapsulated in pumpkin protein isolate by freeze and spray drying method.Methods: The powders were characterised by measuring moisture content, hygroscopicity, bulk density, solubility, a* value (colour). To simulate the human gastrointestinal digestion, in vitro digestion was conducted as two-stage hydrolysis by pepsin and by pancreatin. The antioxidant and ACE inhibitory potential of the digests was examined.Results: Encapsulation efficiency of phenols from beetroot juice in pumpkin protein isolate was 92% for freeze and 75% for spray dried. Physical properties of all samples were affected by the drying methods. All powders were easily digested. It was observed that the antioxidant and ACE inhibitory potential of the digests was enhanced, indicating that it originated from the hydrolysates released from the protein carrier during digestion.Conclusion: The results from this study are promising and indicate that the pumpkin oil cake protein has great potential and could be introduced to the encapsulation process of bioactive compounds as a new carrier agent.

Bioavailability and Bioactivity of Encapsulated Phenolics and Carotenoids Isolated from Red Pepper Waste.

In order to deactivate the health properties of bioactive compounds, they need to withstand the effects of food processing, their potential release from the food matrix, and remain bio-accessible in the gastrointestinal tract. Bio-actives from different plants are prone to oxidative degradation, and encapsulation is an effective method in improving their stability. In the present study, red pepper waste (RPW), a by-product of vegetable processing industry, was encapsulated in whey protein using spray and freeze-drying techniques. The aim was to evaluate the effects of in vitro gastrointestinal digestion on the release and bioactivity of encapsulated bio-actives, after each digestion step. The results showed that the release of phenolics and carotenoids, as well as antioxidants, anti-hyperglycemic, and anti-inflammatory activities are influenced by pH and intestinal fluid, with pH 7.5 exhibited at higher levels. There was a rapid initial release of carotenoids from whey protein matrices, while a more gradual increase of phenolics was observed, reaching around 50% for both encapsulates first at 6 h and 37 °C, and small intestine conditions. The encapsulation of RPW demonstrated a protective effect against pH changes and enzymatic activities along digestion, and contributed to the increase in bio-accessibility in the gut. Also, the results suggest that encapsulation is an efficient method for valorization of bio-actives from RPW, with improvements in nutrition, color, and bioactive properties.

Microencapsulates and extracts from red beetroot pomace modify antioxidant capacity, heat damage and colour of pseudocereals-enriched einkorn water biscuits.

Cereals supply humankind with carbohydrates, proteins and several health-enhancing compounds, including antioxidants. Pomace, a by-product of beetroot juice preparation, is rich in antioxidants (phenolic compounds and betalains). The aim of this work was to study the effect of pomace extract addition, either pure or microencapsulated, on antioxidant properties, heat damage and colour of einkorn water biscuits enriched with pseudocereals. Pomace extract addition had different effects on total polyphenol contents and antioxidant capacity (FRAP and ABTS) in diverse blends. In bread, wheat and einkorn matrices, a significant increase was observed, while in pseudocereals-enriched blends, richer in antioxidants, only microencapsulation improved their content. Pomace extract addition led to furosine reduction and hydroxymethylfurfural increase. Microencapsulate-enriched WB were richest in betanin, isobetanin, total phenolics and antioxidant capacity. In conclusion, pomace extracts, by-products of juice manufacturing, significantly improve some nutritional characteristics of baked products, especially when conveyed as microencapsulates.

Encapsulation of Sour Cherry Pomace Extract by Freeze Drying: Characterization and Storage Stability.

In this study sour cherry pomace was extracted with food-grade solvent (50% ethanol), concentrated and stabilized in whey and soy proteins by encapsulation. Soy proteins exhibited higher encapsulation efficiency (94.90%), but not significantly (p < 0.05), from whey (90.10%). Storage properties of whey (WP) and soy protein (SP) encapsulates in terms of total polyphenols, anthocyanins and antioxidant activity were tested for 6 weeks. At the end of storage period the retention of polyphenols in SP and WP was similar (67.33 and 69.30%, respectively), while the content of anthocyanins has increased in SP (for 47.97%) and decreased in WP (for 1.45%). The decrease in antioxidant activity in SP (12.22%) was lower than in WP (35.04%). Colour parameters of encapsulates have followed the similar trend as anthocyanin change during storage. The technique reported herewith can be used for obtaining quality encapsulates for their use as functional food additives, as a way of fruit waste valorization.

Bioactive Compounds and Antioxidant Properties of Goji fruits (Lycium barbarum L.) Cultivated in Serbia.

OBJECTIVE: Goji fruit extracts, methanol (MGE) and hexane (HGE), were subjected to evaluation as potential source of phenolic antioxidants and antiradical activity. METHODS: Some phenolic compounds (gallic, protocatechuic, vanillic, chlorogenic, coumaric, caffeic and ferulic acid and catechin and rutin), vitamin C and carotenoids were identified and quantified by HPLC. Antioxidant activity was tested by measuring ability to scavenge DPPH and hydroxyl radicals. Also, reducing power of goji fruit extracts was determined. RESULTS: HPLC analysis results showed predominance of gallic acid (40.44 mg/g g.f). Vitamin C content in MGE was 716.91 mg vitC/100 g g.f. IC50DPPH· varied from 26.64 µmolTEAC/g for HGE to 62.15 µmolTEAC/g for MGE, while RP0.5 values varied from 952.23 µmolTEAC/g for MGE to 1360.48 mg/mL for HGE. IC50·OH for MGE was 1844.01 µmolTEAC/g. CONCLUSIONS: Our results support the use of goji fruits as rich sources of phytochemicals for further utilization in the food industry as supplements and functional food ingredients.

Encapsulation of Beetroot Pomace Extract: RSM Optimization, Storage and Gastrointestinal Stability.

One of the great problems in food production are surplus by-products, usually utilized for feeding animals and for preparation of dietary fibre or biofuel. These products represent potential sources of bioactive antioxidants and colour-giving compounds which could be used in the pharmaceutical industry and as food additives. In the present study beetroot pomace extract was encapsulated in soy protein by a freeze drying method. Process parameters (core: wall ratio, extract concentration and mixing time) were optimized using response surface methodology (RSM) in order to obtain the optimum encapsulate (OE) with the highest polyphenol encapsulation efficiency (EE) and radical scavenging activity on DPPH radicals (SA). Using the calculated optimum conditions, the EE (86.14%) and SA (1668.37 µmol Trolox equivalents/100 g) of OE did not differ significantly (p < 0.05) from the predicted ones. The contents of total polyphenols (326.51 mg GAE/100 g), flavonoids (10.23 mg RE/100 g), and betalains (60.52 mg betanin/100 g and 61.33 mg vulgaxanthin-I/100 g), individual content of phenolic compounds and betalains by HPLC, and the ability to reduce Fe(3+) ions, i.e., reducing power (394.95 µmol Trolox equivalents/100 g) of OE were determined as well. During three months of storage at room temperature, polyphenol retention was much higher (76.67%) than for betalain pigments, betacyanins (17.77%) and betaxanthins (17.72%). In vitro digestion and release of phenolics from OE showed higher release rate in simulated intestinal fluid than in gastric fluid. These results suggest encapsulation as a contemporary method for valorisation of sensitive bioactive compounds from food industry by-products.

Sour cherry pomace extract encapsulated in whey and soy proteins: Incorporation in cookies.

One of the potential sources of valuable bioactives is pomace, a by-product from fruit juice processing industry. In the presented study, bioactive compounds extracted from cherry pomace, encapsulated in whey and soy proteins, have been incorporated in cookies, replacing 10% (WE10 and SE10) and 15% (WE15 and SE15) of flour. Total polyphenols, anthocyanins, antioxidant activity and colour characteristics of enriched cookies were followed during 4 months of storage. Total polyphenols of WE10, SE10, WE15 and SE15 have shown a slight increase (23.47, 42.00, 4.12 and 1.16%, respectively), while total anthocyanins (67.92, 64.33, 58.75 and 35.91%, respectively) and antioxidant activity (9.31, 24.30, 11.41 and 12.98%, respectively) decreased. Colour parameters (L(∗), a(∗) and b(∗)) of cookies were influenced by the colour of encapsulates. Fortified cookies received satisfactory sensory acceptance as well. Encapsulated sour cherry pomace bioactives have positively influenced functional characteristics of fortified cookies and their preservation.

Comparative analysis of antioxidant, antimicrobiological and cytotoxic activities of native and fermented chamomile ligulate flower extracts.

MAIN CONCLUSION: The work investigated differences in apigenin content, as well as in other compounds, and examined the chemical profiles, antioxidant, antimicrobial and cytotoxic effects of extracts obtained from native and fermented chamomile ligulate flowers. Chamomile (Chamomilla recutita L.) has a long history of being used as a medicinal plant due to many health benefits, including antiinflammatory, anticancer, antispasmodic, radical-scavenging effects and others. Apigenin is recognized as one of the most bioactive phenolic compounds in chamomile. In comparison to its bound forms, which include mostly apigenin-7-O-ß-glucoside and various acylated forms, the aglycone is attributed with much higher bioactivity. Due to this fact, in this work ligulate florets of chamomile anthodium were subjected to a fermentation process using native chamomile enzymes to hydrolyze bound forms of apigenin to free aglycone. The contents of apigenin and apigenin-7-O-ß-glucoside were determined in both fermented and nonfermented samples by UHPLC-MS-MS analysis to define the efficiency of conversion. After defining their chemical profiles, the extracts of fermented and nonfermented chamomile samples were also compared with respect to their antioxidant, antimicrobial and cytotoxic effects. The antioxidant effects of the obtained extracts were defined by electron spin resonance analysis for hydroxyl and superoxide radicals. The antimicrobial activity was defined for eight microbial strains, whereas cytotoxic activity was evaluated using two human cell lines (human cervix carcinoma and human rhabdomyosarcoma) and murine fibroblasts.

Tomato waste: Carotenoids content, antioxidant and cell growth activities.

The carotenoid content, antioxidant and cell growth activities of tomato waste extracts, obtained from five different tomato genotypes, was investigated. High performance liquid chromatography was used to identify and quantify the main carotenoids present in tomato waste extracts. The antioxidant activity of tomato waste extracts was tested using spectrophotometric methods, 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity and reducing power assay. The highest DPPH scavenging activity (IC50 = 0.057 mg/ml) was obtained for Backa extract. The Knjaz extract showed the best reducing power (IC50 = 2.12 mg/ml). Cell growth effects were determined in HeLa, MCF7 and MRC-5 cell lines by sulforhodamine B test. Anti-proliferative effects were observed in all cell lines at higher concentrations (⩾ 0.125 mg/ml). The carotenoid contents exhibited a strong correlation with antioxidant and anti-proliferation activity. The results obtained indicated that tomato waste should be regarded as potential nutraceutic resource and may be used as a functional food ingredient.

Bioactivity of Meeker and Willamette raspberry (Rubus idaeus L.) pomace extracts.

Taking into account the substantial potential of raspberry processing by-products, pomace extracts from two raspberry cultivars, Meeker and Willamette, were investigated. Total phenolic, flavonoid and anthocyanin contents were determined. Willamette pomace extract (EC50=0.042 mg/ml) demonstrated stronger 2,2-diphenyl-1-picrylhydrazyl DPPH radical-scavenging activity than did Meeker pomace extract (EC50=0.072 mg/ml). The most pronounced cell growth inhibition effect was obtained in the breast adenocarcinoma cell line, reaching EC50 values of 34.8 and 60.3 µg/ml for Willamette and Meeker extracts, respectively. Both extracts demonstrated favourable non-tumor/tumor cell growth ratios and potently increased the apoptosis/necrosis ratio in breast adenocarcinoma and cervix carcinoma cells. In reference and wild bacterial strains, minimal inhibitory concentrations (MIC) were achieved in a concentration range from 0.29 to 0.59 mg/ml, and minimal bactericidal concentrations (MBC) in a range from 0.39 to 0.78 mg/ml. The results indicate significant antioxidant, antiproliferative, proapoptotic and antibacterial activities of raspberry pomace and favour its use as a functional food ingredient.

Antiradical, antimicrobial and cytotoxic activities of commercial beetroot pomace.

The by-product of food processing is often utilized as feed, and for the preparation of dietary fiber and biofuel. However, these products are also promising sources of bioactive antioxidants and color giving compounds, which could be used as additives in the food, pharmaceutical and cosmetic industry. The aim of this study was to investigate the phytochemical profile, and the antiradical, antimicrobial and cytotoxic activities of industrial beetroot pomace extract (BPE). The content of phenolics (45.68 mg gallic acid equivalents g(-1)), flavonoids (25.89 mg rutin equivalents g(-1)) and betalains (4.09 mg betanin g(-1); 7.32 mg vulgaxanthin I g(-1)) were determined spectrophotometrically. The antiradical activity on DPPH (EC(50)(DPPH·) = 0.0797 mg ml(-1)), hydroxyl (EC(50)(·OH) = 0.0655 mg ml(-1)) and superoxide anion (EC(50)(O2·-) = 1.0625 mg ml(-1)) radicals were measured by electron spin resonance (ESR) spectroscopy. The antimicrobial activity was determined using the agar-well diffusion method. Gram(-) bacteria (Salmonella typhimurium, Citrobacter freundii) and Gram(+) bacteria, (Staphylococcus aureus, Staphylococcus sciuri, Bacillus cereus) showed high susceptibility, while yeasts and moulds were resistant. BPE exhibits cytotoxic properties against Ehrlich carcinoma (EAC) cells in vivo due to induction of oxidative stress. The largest decreases in EAC cell numbers were observed in the pre-treated male (approximately 53%) and female (approximately 47%) mice, and also the EAC cell viability was decreased after administration of BPE. The activities of the antioxidant enzymes, xanthine oxidase (XOD) and peroxidase (Px), were significantly different between the untreated EAC control group and all other groups that were treated with BPE. The XOD and Px activities were very low in untreated malignant cells, but increased significantly after administration of BPE. Our results show that BPE holds promise in the food industry as a source of bioactive compounds.

Effect of rosehip (Rosa canina L.) phytochemicals on stable free radicals and human cancer cells.

BACKGROUND: The commercial development of plants as sources of antioxidants that can be used to enhance the properties of foods, for nutritional purposes and preservation as well as for prevention of oxidation-related diseases, is currently of major interest. Rosehip (Rosa canina L.) is a rich source of vitamin C and polyphenols. RESULTS: Phytochemicals in rosehip tea were separated into three fractions: Fr1 (vitamin C, 39.17 mg kg(-1)), Fr2 (flavonoids, 451.05 µg kg(-1)) and Fr3 (phenolic acids, 504.69 µg kg(-1)). Quercetin and ellagic acid were the most abundant polyphenolic compounds. Rosehip fractions, primarily rosehip flavonoids (EC(50) = 49 mg L(-1)), showed high antioxidant activity towards 2,2-diphenyl-1-picrylhydrazyl radicals (DPPH(•)). Cell growth effects of rosehip fractions were assessed in HeLa, MCF7 and HT-29 cell lines, with the lowest IC(50) values being determined for rosehip flavonoids, (80.63, 248.03 and 363.95 mg L(-1) respectively). However, the vitamin C fraction did not inhibit the growth of tested tumour cells. CONCLUSION: The results of this study confirm that vitamin C and flavonoids are responsible for the antioxidant activity of rosehip tea, while only polyphenols contribute to its antiproliferative activity.

Tomato waste: a potential source of hydrophilic antioxidants.

Waste obtained from six genotypes of tomato was subjected to evaluation as potential sources of hydrophilic antioxidants on the basis of content of total phenolics (TPh; from 11.7 to 18.6 mg/g), total flavonoids (TFl; from 7.62 to 12.1 mg/g) and ascorbic acid (AA; from 0.51 to 1.89 mg/g), and antioxidant activities. The antioxidant activities of tomato waste extracts were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, ferrous ion chelating activity (CA) and reducing power (RP) assay. The O(2) tomato waste extract was found to have the strongest antioxidant activity in scavenging DPPH radicals (effective concentration, EC(50) = 0.18 mg/ml). The Saint Pierre tomato waste extract showed the best performance in RP assay (EC(50) = 1.57 mg/ml). The Rutgers tomato waste extract exhibited a strongest CA (EC(50) = 1.49 mg/ml). The linear correlation analysis produced moderate-to-high correlation coefficients between the antioxidant activities and TPh, TFl and AA.

Antioxidative and antiproliferative activities of different horsetail (Equisetum arvense L.) extracts.

In this study we investigated antioxidative and antiproliferative activity of different horsetail (Equisetum arvense L.) extracts. The antioxidative activity was measured by the electron spin resonance (ESR) spectroscopy-spin trapping method. The influence of different horsetail extracts during lipid peroxidation of (1) sunflower oil induced by the lipophilic azo-initiator 4,4'-azobis(4-cyanovaleric acid) and (2) soybean phosphatidylcholine liposomes induced by the hydrophilic azo-initiator 2,2'-azobis(2-amidinopropane) dihydrochloride was studied. Antiproliferative activity was measured using the sulforhodamine B colorimetric assay on the human cancer cell lines HeLa, HT-29, and MCF7. The results of ESR analysis confirmed that the extracts investigated suppressed the formation of lipid peroxyl radicals in both systems investigated in a dose-dependent manner. The results indicate that n-butanol, methanol, ethyl acetate, and water extracts had significant peroxyl radical scavenging activity. Extracts inhibited cell growth that was dependent on cell line, type of extract, and extract concentration. Ethyl acetate extract exhibited the most prominent antiproliferative effect, without inducing any cell growth stimulation on human tumor cell lines. The results obtained suggest that the horsetail extracts could be used as an easily accessible source of natural antioxidants and as potential phytochemicals.

Radical scavenging, antibacterial, and antiproliferative activities of Melissa officinalis L. extracts.

The aromatic herb Melissa officinalis L. can be used as an easily accessible source of natural antioxidants and as a possible food supplement and as a phytochemical. Radical scavenging, antibacterial, and antiproliferative activities of petroleum ether, chloroform, ethyl acetate, n-butanol, and water extracts of M. officinalis L. extracts were investigated. The results of antioxidative activity, obtained by electron spin resonance spectroscopy, confirmed that investigated extracts suppressed the formation of 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl, and lipid peroxyl radicals in all investigated systems in a dose-dependent manner. The maximum DPPH and hydroxyl radical scavenging activities (SA(DPPH) = SA(OH) = 100%) were achieved in the presence of n-butanol extract at concentrations of 0.4 mg/mL and 0.5 mg/mL, respectively. The highest lipid peroxyl scavenging activity (93.20%) was observed at a higher concentration (5 mg/mL) of n-butanol extract in the lipid peroxidation system. The most effective antibacterial activities were expressed by petroleum ether and ethyl acetate extracts on Sarcina lutea. Chloroform extract showed the strongest antiproliferative effect with 50% inhibitory concentration values of 0.09 mg/mL and 0.10 mg/mL for HeLa and MCF-7 cell lines, respectively. The present study demonstrated the high phenolic content and radical scavenging, antibacterial, and antiproliferative activities of extracts of M. officinalis L. originating from Serbia.

Antimicrobial and free radical scavenging activities of Teucrium montanum.

The antimicrobial and free radical scavenging activities of petroleum ether, chloroform, ethyl acetate, n-butanol and water extracts of Teucrium montanum were investigated. Ethyl acetate, chloroform and n-butanol extracts expressed a wide range of inhibiting activity against both Gram (+) and Gram (-) bacteria. n-Butanol extract possessed potent DPPH free radical scavenging activity.