Contribution of Insoluble Bound Antioxidants from Germinated Seeds of Wheat and Spelt to the Nutritional Value of White Bread.

This research aimed to study the dynamic changes in phenolic antioxidants between the germ and the rest of the germinated seed of wheat and spelt and to evaluate the whole grain flour from germinated seeds as a functional supplement. Longer germination resulted in higher TPC, DPPH, and ABTS values when considering the entire germinated seed, while the optimal germination time was not consistent when considering the germ and the remaining germinated seed separately. While in the germinated seed (without germ) the majority of TPC was determined to be bound phenolics (up to 92%), the extractable form dominated in the germ (up to 69%). The most abundant phenolic antioxidants in germinated wheat and spelt seeds, trans-ferulic acid, cis-ferulic acid, and p-coumaric acid, increased significantly with germination. Only breads with 5% germinated spelt or wheat flour were suitable for the production of a food product, showing higher extractable TPC, antioxidant activity, individual phenolic acids, and improved specific volume, and were preferred because of their appearance, aroma, and color. The PCA biplot showed that the addition of 15% and 30% germinated flours had the greatest positive impact on phenolic properties, while breads with the addition of 5% germinated flour had the greatest positive impact on specific volume and color.

Bioprocessed Wholegrain Spelt Flour Improves the Quality and Physicochemical Characteristics of Wheat Bread.

In the present study, the partial substitution of common white wheat flour for a bread recipe with variously bioprocessed wholegrain spelt was investigated. The addition of 1% and pasteurised 5% "germinated + enzymatic treated" spelt flour to wheat flour significantly improved the specific volume of the bread, but their texture profile analysis and sensory evaluation were not satisfactory. A higher percentage of added bioprocessed spelt flour darkened the colour of the bread. Breads with the addition of more than 5% of bioprocessed spelt flour were unacceptable in terms of quality and sensory parameters. The highest extractable and bound individual phenolics were found in breads with 5% "germinated + fermented" spelt flour (GFB5) and 5% pasteurised "germinated + enzymatic treated" spelt flour (GEB5P). A strong positive correlation was determined between trans-ferulic acid and TPC and DPPH• radical scavenging activity. The GEB5P bread showed the highest increase in extractable and bound trans-ferulic acid content, by 320% and 137%, respectively, compared to the control bread. Principal component analysis showed differences between the control bread and enriched breads in terms of their quality, sensory and nutritional properties. Breads with 2.5% and 5% "germinated + fermented" spelt flour had the most acceptable rheological, technological and sensory characteristics, in addition to a substantial improvement in their antioxidant content.

Changes in the Bioaccessibility of Antioxidants after Simulated In Vitro Digestion of Bioprocessed Spelt-Enhanced Wheat Bread.

The aim of the study was to determine whether the partial replacement of wheat flour with bioprocessed spelt flour contributes to a higher bioaccessibility of the antioxidants in bread. The results showed that the type and amount of bioprocessed spelt flour in a bread recipe has a major impact on the extractable and bound TPC, the content of individual phenolics, their antioxidant activity, and their bioaccessibility as determined by in vitro digestion. Extractable p-coumaric and trans-ferulic acids in breads decreased after digestion, while extractable cis-ferulic and p-hydroxybenzoic acids increased. The bioaccessibility of TPC in the control bread (100% wheat flour), and in bread enriched with 5% "germinated + fermented" spelt flour (GFB5), did not differ. However, the digested GFB5 bread contained 5.2-times more extractable, and 1.3-times more bound, trans-ferulic acid than the digested control bread. trans-Ferulic acid showed the lowest bioaccessibility, up to 2.8%. In GFB2.5 and GFB5 breads, the bioaccessibility of p-coumaric, trans-ferulic, and cis-ferulic acids was higher than in other digested breads. PCA visualized the difference between the undigested and digested breads. The incorporation of germinated and fermented, or germinated and enzymatic, treated spelt flour in a white bread recipe could be an attractive way of providing consumers with nutritionally interesting foods.

Simulated Gastrointestinal Digestion of Bioprocessed Spelt Seeds: Bioaccessibility and Bioactivity of Phenolics.

The goal of this research was to evaluate the impact of different bioprocessing techniques on improved bioaccessibility of phenolics from spelt seeds. Despite the negative influence of gastrointestinal digestion, fermentation of germinated seeds significantly increased the bioaccessibility of total phenolics and their antioxidant activity compared to digested raw seeds. Enzymatic treated fermented seeds showed the highest relative bioaccessibility of p-coumaric and trans-ferulic acids, while their absolute contents were significantly higher in "germinated + fermented" seeds. Our research suggests that pretreatment of spelt seeds with hydrolytic enzymes improves access of fermenting microorganisms to structural elements, resulting in an increased content of extractable and bound trans-ferulic acid. Significantly higher biostability of phenolics was observed in raw seeds. Some major quality changes in the composition of extracts were observed under simulated in vitro digestion, since antioxidants of the same extract showed a different relative decrease in DPPH• and ABTS•+ scavenging activities compared to the raw seeds or their corresponding undigested counterparts. It is therefore important to increase the content of extractable antioxidants in seeds by bioprocessing, since they are strongly diminished during digestion.

Improving accessibility and bioactivity of raw, germinated and enzymatic-treated spelt (Triticum spelta L.) seed antioxidants by fermentation.

Lactic acid, alcoholic, combined and spontaneous fermentation of raw, germinated and enzymatic-treated spelt seeds significantly improved the content of extractable and bound phenolics and considerably increased the extractable:bound ratio, and therefore positively affected the accessibility of the spelt antioxidants. The highest extractable and bound individual phenolic contents and in vitro antioxidant activities of extracts were obtained following fermentation of germinated spelt seeds with Saccharomyces cerevisiae, while for enzymatic-treated seeds, Lactobacillus plantarum (alone or with S. cerevisiae) was the most effective. For extractable phenolics, trans-ferulic acid increased the most in yeast-fermented germinated seeds (2922%); for bound phenolics, cis-ferulic acid showed the greatest relative increase in yeast-fermented raw spelt seeds (466%). Spontaneous fermentation of germinated and enzymatic-treated samples decreased intracellular oxidation most effectively, probably due to apigenin derivatives. Cellular uptake of bound hydroxycinnamic acids was significantly higher than that of extractable hydroxycinnamic acids; however, the latter were more efficient in vivo antioxidants.

Development and Optimisation of Solid-Phase Extraction of Extractable and Bound Phenolic Acids in Spelt (Triticum spelta L.) Seeds.

A solid-phase extraction (SPE) technique was developed and optimised for isolation and concentration of extractable and bound phenolic acids from germinated spelt seeds, for analysis by liquid chromatography-mass spectrometry. Samples initially underwent solvent extraction under different conditions to maximise the yield of phenolic antioxidants. Optimal extraction conditions for extractable phenolics were absolute methanol as solvent, sample-to-methanol ratio 1:9, and reconstitution in non-acidified water. The bound phenolics were extracted from sample pellets using hydrolysis with 2 M NaOH, acidification of the hydrolysate with formic acid, and simultaneous isolation and purification using Strata X polymeric RP tubes. Compared to liquid-liquid extraction, this direct SPE protocol has significant advantages in terms of higher extraction efficiencies of total and individual phenolics and their antioxidant activities. These data suggest that direct SPE represents a rapid and reliable method for quantitative analysis of both the extractable and the commonly overlooked bound phenolics in Triticum spelta seeds.

Abiotic stress combinations improve the phenolics profiles and activities of extractable and bound antioxidants from germinated spelt (Triticum spelta L.) seeds.

The aim of this study was to investigate the effects of germination of spelt seeds under different stress conditions on the antioxidant characteristics of their extractable and bound phenolics. Germination under combined stress of 25 mM NaCl and 50 mM sorbitol without subsequent mechanical stress had considerable impact on total phenolics contents and scavenging activities against different free radicals (DPPH, ABTS+, O2-, ROO). Alkaline hydrolysis of extracts from germinated seeds provided the majority of their phenolic acids, where ferulic and p-coumaric acids were the most representative. The phenolics liberated from their bound form also had greater antioxidant activities. For the extractable phenolics, p-coumaric hexoside increased the most (146%), while among the bound phenolics identified, the highest relative increase was for p-coumaric acid (171%). The germinated seeds showed no effects on intracellular oxidation in cells of the yeast Saccharomyces cerevisiae.

LC-MS analysis of phenolic compounds and antioxidant activity of buckwheat at different stages of malting.

The impact of malting on the profile of the phenolic compounds and the antioxidant properties of two buckwheat varieties was investigated. The highest relative increases in phenolic compounds were observed for isoorientin, orientin, and isovitexin, which are consequently major inducible phenolic compounds during malting. Only a minor relative increase was observed for the most abundant phenolic compound, rutin. The radical-scavenging activity of buckwheat seeds was evaluated using ABTS and DPPH assays. A considerable increase in total phenolic compounds and higher antioxidant activity were observed after 64h of germination, whereas kilning resulted in decreased total phenolic compounds and antioxidant activity. Higher antioxidant activities for extracts were found for buffered solvents than for pure methanol and water. Changes in the composition of the phenolic compounds and increased antioxidant content were confirmed by several methods, indicating that buckwheat malt can be used as a food rich in antioxidants.

Effect of deodorization of camelina (Camelina sativa) oil on its phenolic content and the radical scavenging effectiveness of its extracts.

The influence of deodorization parameters (temperature (T), steam flow (S), time (t)) on the phenolic content and radical scavenging effectiveness (RSE) of methanolic extracts of camelina oil was investigated and analyzed by response-surface methodology (RSM). The phenolic content can be considered to be a linear function of all three parameters. A positive linear relationship between the content of phenolic compounds in deodorized oils and RSE was observed. Deodorization at 210 °C with a steam flow of 3 mL/h for 90 min resulted in the best preservation of phenolics, amounting to 29.9 mg/kg. The lowest reduction from RSE of 12.4 µM Trolox equivalents (TE)/g oil for the crude oil was observed for oil treated at 195 °C and 18 mL/h for 60 min with RSE of 10.1 µM TE/g oil. The lack of correlation between RSE or total phenolic content and oxidative stability (OS) of the deodorized oils suggests that antioxidants in scavenging radicals react by different mechanisms, depending on radical type and reaction medium.

Effect of heat treatment of camelina (Camelina sativa) seeds on the antioxidant potential of their extracts.

The effect of different heat treatments of camelina (Camelina sativa) seeds on the phenolic profile and antioxidant activity of their hydrolyzed extracts was investigated. The results showed that total phenol contents increased in thermally treated seeds. Heat treatment affected also the quantities of individual phenolic compounds in extracts. Phenolics in unheated camelina seeds existed in bound rather than in free form. A temperature of 160 °C was required for release of insoluble bound phenolics, whereas lower temperatures were found to be optimal to liberate those present as soluble conjugates. The best reducing power and alkyl peroxyl radical scavenging activity in the emulsion was expressed by phenolics which were bound to the cell wall, whereas the best iron chelators and 2,2-diphenyl-1-picrylhydrazyl (DPPH•) radical scavengers were found to be those present in free form. The heat treatment of seeds up to 120 °C increased the reducing power and DPPH• radical scavenging ability of extracts, but negatively affected iron chelating ability and their activity in an emulsion against alkyl peroxyl radicals.

Antioxidant properties of 4-vinyl derivatives of hydroxycinnamic acids.

The compounds 4-vinylphenol (4-VP), 4-vinylguaiacol (4-VG), 4-vinylsyringol (4-VS) and 4-vinylcatechol (4-VC) were prepared by thermal decarboxylation of the corresponding hydroxycinnamic acids p-coumaric, ferulic, sinapic and caffeic acid, respectively. For confirmation of the synthesised products LC-MS followed by NMR analysis was used. To evaluate their antioxidant potential, their reducing power and efficiency in scavenging the alkylperoxyl radical generated in an emulsion system, the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical and the superoxide anion radical (O2(-)) were determined. All tested 4-vinyl derivatives revealed weaker antioxidant activity in a homogeneous polar medium than the corresponding phenolic acids. In the emulsion system the activity for 4-vinyl derivatives was higher than was the activity of their corresponding phenolic acids, with 4-VG as the most active among the tested phenolic compounds.