Cranberry and black chokeberry extracts isolated with pressurized ethanol from defatted by supercritical CO2 pomace inhibit colorectal carcinoma cells and increase global antioxidant response of meat products during in vitro digestion.

Berry phenolics are considered as phytochemicals, which might mitigate development of degenerative diseases, including cancer. Many studies demonstrated their antiproliferative effects in various cancer cell lines while the studies with real foods are rather scarce. We report antiproliferative properties of unique extracts, which were obtained from the defatted by supercritical CO2 cranberry (CrE) and black chokeberry (ChoE) pomace using pressurized ethanol, and global antioxidant response of meat products enriched with berry polyphenolics during in vitro digestion. ChoE was more effective against HCT116 and DLD1 cells than CrE, while the HCT116 cells were more sensitive to digested meat samples than DLD1. At 1000 µL ChoE reduced cell viability to 51% (HCT116) and ∼50% (DLD1), while in case of CrE >69% of HCT116 cells remained viable. The extracts added at 2% increased antioxidant capacity values of hamburgers and cooked ham at oral and gastric digestion phases; however, at intestinal phase no regular effects were observed. The highest antioxidant potential was determined in hamburgers/cooked ham with 2% of CrE (TPC: 1.45/2.01 mg GAE/mL; ABTS●+: 9.82/15.66 mg TE/mL; ORAC: 13.58/12.08 mg TE/mL). The content of quantifiable anthocyanins remarkably decreased in the digesta at all phases and particularly at intestine phase: >99% with CrE and 97-99% with ChoE. Digested liquids of cooked ham prepared with extracts significantly stronger inhibited HCT116 cells at selected dilution factors. The results obtained provide preliminary information that cranberry and black chokeberry pomace extracts may provide health benefits when added in meat products.

Recovery of Bioactive Compounds from Strawberry (Fragaria × ananassa) Pomace by Conventional and Pressurized Liquid Extraction and Assessment Their Bioactivity in Human Cell Cultures.

Pressing strawberries for juice generates large amounts of pomace, containing valuable nutrients and therefore requiring more systematic studies for their valorization. This study compared conventional solid-liquid (SLE) and pressurized liquid (PLE) extractions with ethanol (EtOH) and H2O for the recovery of bioactive compounds from strawberry pomace. The composition and bioactivities of the products obtained were evaluated. Among 15 identified compounds, quercetin-3-glucuronide, kaempferol-3-glucuronide, tiliroside, ellagic, malic, succinic, citric and p-coumaric acids were the most abundant constituents in strawberry pomace extracts. SLE-EtOH and PLE-H2O extracts possessed strong antioxidant capacity in DPPH• and ABTS•+ scavenging and oxygen radical absorbance capacity (ORAC) assays. Cytotoxicity, antiproliferative and cellular antioxidant activities in human cells of PLE-EtOH and PLE-H2O extracts were also evaluated. PLE-EtOH and PLE-H2O extracts possessed strong antioxidant activity, protecting Caco-2 cells upon stress stimuli, while PLE-EtOH extract showed higher antiproliferative activity with no cytotoxicity associated. In general, the results obtained revealed that properly selected biorefining schemes enable obtaining from strawberry pomace high nutritional value functional ingredients for foods and nutraceuticals.

Valorization of European Cranberry Bush (Viburnum opulus L.) Berry Pomace Extracts Isolated with Pressurized Ethanol and Water by Assessing Their Phytochemical Composition, Antioxidant, and Antiproliferative Activities.

Defatted by supercritical CO2, Viburnum opulus berry pomace (VOP) was subjected to consecutive extraction with pressurized ethanol (E) and water (W) and yielded 23% of VOP-E and 8% of VOP-W, respectively. The major phytochemical groups covering 42 identified and quantified constituents in VOP extracts were organic and phenolic acids, iridoids, quercetin and (epi)catechin derivatives, flavalignans, procyanidins, and anthocyanins. The on-line HPLC-DPPH•-scavenging assay revealed the presence of numerous antioxidants. VOP-E had a higher total phenolic content, was a stronger antioxidant (equivalent to 0.77, 0.42, and 0.17 g trolox/g in oxygen radical absorbance capacity (ORAC), ABTS, and DPPH assays, respectively), and recovered the major part of phenolics from the pomace; however, both extracts demonstrated similar antioxidant activity in the cellular assay. VOP-E inhibited HT29 cancer cells at non-cytotoxic concentrations. The results of this study revealed that VOP contains valuable phytochemicals possessing antioxidant and antiproliferative activities. Consequently, extracts from VOP substances may be of interest in developing functional ingredients for healthy foods, nutraceuticals, and cosmeceuticals.

Zero waste biorefining of lingonberry (Vaccinium vitis-idaea L.) pomace into functional ingredients by consecutive high pressure and enzyme assisted extractions with green solvents.

Due to the lack of innovative valorization strategies, berry pomaces are a poorly utilized as a cheap source of valuable nutrients and phytochemicals. An effective biorefining scheme was developed to recover functional components from lingonberry pomace by consecutive supercritical CO2 (SFE-CO2), pressurized liquid (PLE) and enzyme assisted (EAE) extractions. SFE-CO2 at optimized parameters yielded 11.8 g/100 g of lipophilic fraction, containing 43.3 and 37.4% of α-linolenic and linoleic fatty acids, respectively. The combined PLE with ethanol and water additionally recovered 61.8 g/100 g of polar constituents and reduced the antioxidant capacity of starting material by up to 94%. The major portion of the antioxidants (89-94% in different assays), anthocyanins (231 mg/100 g pomace) and proanthocyanidins (15.9 g/100 g pomace) was present in PLE-EtOH extract. Cyanidin-3-galactoside was the major anthocyanin (146.9 mg/100 g). High-pressure fractionation was more efficient for obtaining bioactive pomace constituents as compared with conventional and enzyme-assisted extractions.

Phytochemical Composition, Antioxidant and Antiproliferative Activities of Defatted Sea Buckthorn (Hippophaë rhamnoides L.) Berry Pomace Fractions Consecutively Recovered by Pressurized Ethanol and Water.

This study aimed at valorisation of sea buckthorn pomace (SBP) for the production of extracts containing valuable bioactive compounds. For this purpose, SBP defatted by supercritical CO2 was subjected to consecutive fractionation with pressurized ethanol and water, which yielded 11.9% and 4.8% of extracts, respectively. The extracts were evaluated for their antioxidant potential, phytochemical composition and antiproliferative effects against cancer cells. Water extracts exhibited remarkably higher values in Folin-Ciocalteu assay of total phenolic content, oxygen radical absorbance capacity (ORAC), ABTS●+/DPPH scavenging and cellular antioxidant activity (CAA) assays and more efficiently inhibited proliferation of HT29 cells at non-cytotoxic concentrations measured in non-tumoral Caco2 cells. Among 28 detected and 21 quantified phytochemicals, flavonols with the structures of isorhamnetin (five compounds), quercetin (three compounds), kaempferol (three compounds) glycosides and catechin (six compounds) were the most abundant in the extracts. In conclusion, the applied method of fractionation of SBP produces promising natural antioxidant complexes with antiproliferative properties that could find potential applications in nutraceuticals, functional foods and cosmeceuticals.

Freeze-drying of black chokeberry pomace extract-loaded double emulsions to obtain dispersible powders.

Black chokeberry pomace extract is rich in polyphenolic antioxidants, including anthocyanins. Added to foods, bioactive compounds of the extract can undergo undesirable changes both during food handling and digestion. In this study, we examined the possibility of encapsulating a considerable amount of black chokeberry pomace extract in the inner water phase of double emulsion (water-in-oil-in-water), for intended use in food applications. Furthermore, this study investigated the feasibility of double emulsions loaded with the extract for freeze-drying to obtain dispersible powders. A substantial amount (2.1%) of black chokeberry pomace extract was efficiently encapsulated in the inner water phase of double emulsion and remained entrapped during 60 days of storage (<97%) as well as during the freeze-drying of emulsions. Reconstituted emulsions obtained after the rehydration process were found to show monomodal droplet size distribution, decent creaming stability (approximately 97%), and good encapsulation efficiency (95.36%). Such characteristics of powdered double emulsions loaded by black chokeberry pomace extract make them suitable for food application as retainer and preservative of bioactive polyphenolic-rich extracts. PRACTICAL APPLICATION: Powders of double emulsions loaded by black chokeberry pomace extract could be used as a source of bioactive polyphenolic compounds.

Consecutive high-pressure and enzyme assisted fractionation of blackberry (Rubus fruticosus L.) pomace into functional ingredients: Process optimization and product characterization.

In this study supercritical carbon dioxide (SFE-CO2) and pressurized liquid (PLE) extractions were optimized for the recovery of valuable fractions from blackberry pomace. Consecutively applied SFE-CO2 and PLE at optimized parameters yielded 9.9, 26.3 and 5.1 g/100 g of CO2, ethanol (EtOH) and water-soluble extracts, respectively. Oil of lipophilic fraction was composed mainly of healthy polyunsaturated fatty acids (linoleic 64.1%, α-linolenic 12.9%), while polar solvents effectively recovered antioxidants (up to 29.1 mg gallic acid and 168.7 mg Trolox equivalents from g pomace). PLE-EtOH extract contained 12.2 mg/g of cyanidin-3-glucoside, while other anthocyanins were detected in significantly lower quantities (0.5-0.7 mg/g). SFE-CO2 and PLE reduced the antioxidant capacity of starting plant material by 86-93%. In terms of extraction time, solvent consumption, total yields, and phytochemical characteristics, high-pressure fractionation was more efficient for obtaining valuable pomace constituents as compared to conventional and enzyme-assisted extractions.

Isolation of Strong Antioxidants from Paeonia Officinalis Roots and Leaves and Evaluation of Their Bioactivities.

Paeonia officinalis extracts from leaves and roots were tested for their antioxidant potential using in vitro chemical (Folin-Ciocalteu, 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), oxygen radical absorbance capacity (ORAC), hydroxyl radical antioxidant capacity (HORAC), hydroxyl radical scavenging capacity HOSC)) and cellular antioxidant activity (CAA) assays. Leaf extracts were stronger antioxidants than root extracts, while methanol was a more effective solvent than water in chemical assays. However, the selected water extract of leaves was a stronger antioxidant in CAA than the methanol extract (0.106 vs. 0.046 µmol quercetin equivalents/mg). Twenty compounds were identified by ultra performance liquid chromatography-quadrupole-time-of-flight (UPLC-Q-TOF) mass spectrometer, while on-line screening of their antioxidant capacity by high performance liquid chromatography (HPLC) with a DPPH•-scavenging detector revealed that gallic acid derivatives are the major peony antioxidants. Root water and leaf methanol extracts inhibited α-amylase in a dose dependent manner. The IC50 value for the strongest inhibitor, the methanol extract of leaves, was 1.67 mg/mL. In addition, the cytotoxicity assessment of extracts using human Caco-2 cells demonstrated that none of them possessed cytotoxic effects.

Effect of cranberry pomace extracts isolated by pressurized ethanol and water on the inhibition of food pathogenic/spoilage bacteria and the quality of pork products.

Ethanol and water extracts were prepared from defatted cranberry pomace by pressurized liquid extraction and tested in bacterial cultures of L. monocytogenes, B. thermospacta, P. putida, lactic acid bacteria (LAB), aerobic mesophilic bacteria (AMB), and pork meat products. Anthocynanins (glucosides, galactosides and arabinosides of cyanidin and peonidins), phenolic compounds and organic acids (quinic, chlorogenic, malic and citric acids; procyanidin B3, myricetin and quercetin derivatives) were determined in the extracts. The extracts effectively inhibited the growth of tested bacteria at higher than 3.3% concentration. The effect of 2% ethanol extract additive on the inhibition of the same bacteria was also determined in non-inoculated and inoculated with bacteria pork slurry, pork burgers, and cooked ham. The results showed a significant growth inhibition of pathogenic L. monocytogenes and some other species in pork slurry, burgers and cooked ham with cranberry pomace ethanol extract as compared with the control samples. The extract also effectively inhibited the formation of oxidation indicator malondialdehyde in meat products. Slight impact of extract on some physico-chemical properties of meat products such as pH, metmyoglobin content was also observed, while it did not have significant influence on water activity. Extract addition imparted some color changes; however, it did not have negative effect on the overall sensory quality of burgers and cooked ham. High effectiveness of extract additive against pathogenic L. monocytogenes and some other tested bacteria in pork slurry, burgers and cooked ham during refrigerated storage for 16, 16 and 40 days, respectively, suggest that ethanol extract of defatted cranberry pomace may be a promising natural ingredient of meat products for increasing their microbiological safety and improving oxidative stability.

Genotoxicity and antioxidant activity of five Agrimonia and Filipendula species plant extracts evaluated by comet and micronucleus assays in human lymphocytes and Ames Salmonella/microsome test.

The species of Agrimonia and Filipendula have been traditionally used in folk medicine as anti-inflammatory herbs. This study extends the knowledge on bioactivities of F. palmata, A. eupatoria, A. procera, F. ulmaria and F. vulgaris by comprehensive characterization of their methanolic extracts. Antioxidant properties of extracts were evaluated by DPPH• (2,2-diphenyl-1-picrylhydrazyl), ABTS•+ 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) scavenging and oxygen radical absorbance capacities (ORAC). Genotoxicity of extracts was tested using alkaline single-cell gel electrophoresis (comet) and cytokinesis-block micronucleus assays in human lymphocytes in vitro and the Ames Salmonella/microsome test. All investigated Agrimonia and Filipendula extracts possessed strong antioxidant activity, which was comparable with that of a standard antioxidant trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid). Thirty five compounds belonging to the classes of phenolic acids, flavonoids, phenylpropanoids and ellagitanins were detected by ultra-performance liquid chromatography - mass spectrometry (UPLC-Q-TOF-MS). Agrimonia and Filipendula extracts induced an increase in a DNA damage in the comet assay expressed as mean percentage of DNA in the comet tail. However, these extracts did not produce reverse mutation in bacterial cells in the Ames test and were not genotoxic in the micronucleus test. However, a slight though significant decrease of nuclear division index values was determined. In general, this study proved that Agrimonia and Filipendula species are a good source of bioactive compounds; their extracts may be classified as non-mutagenic and non-clastogenic in vitro under conditions of the current study. Consequently, the plants may be a promising material for nutraceuticals and natural medicines.

Preparation and properties of propylene oxide and octenylsuccinic anhydride modified potato starches.

Starch was dually chemically modified for developing food-grade ingredients of lower digestibility and their properties were compared to those of single modified and native starches. Hydroxypropylation with propylene oxide (HP) followed by esterification with octenylsuccinic anhydride (OSA) of potato starch (P-native) produced derivatives with lower digestibility than esterification solely with OSA. The dextrose equivalent, maltose and glucose contents, which were used as the main indicators for in vitro digestion, were lower for modified starches. P-HP0.2-OSA0.0200 derivative was the least digestible; the glucose and maltose contents were lowered by 28.3 and 42.1% compared to P-native. The aggregation behavior of enzymatically hydrolyzed starch derivatives was studied in aqueous solution by employing the fluorescence probe and dynamic light scattering techniques. The critical aggregation concentration for OSA modified and dually modified starches varied from 1.2 to 3 g/L and from 0.125 to 0.48 g/L, respectively, depending on the degree of OSA substitution. The study showed that above a critical concentration the hydrolyzates of modified starches tend to form the aggregates with different properties depending both on the degree of OSA substitution and chemical structure.