Improving the Extraction of Polyphenols from Cocoa Bean Shells by Ultrasound and Microwaves: A Comparative Study.

The extraction of bioactive compounds from food by-products is one of the most important research areas for the nutraceutical, pharmaceutical, and food industries. This research aimed to evaluate the efficiency of Ultrasound-Assisted Extraction (UAE) and Microwave-Assisted Extraction (MAE), either alone or in combination, of phenolic compounds from cocoa bean shells (CBSs). These extraction techniques were compared with conventional methods, such as under simple magnetic stirring and the Soxhlet apparatus. After the preliminary characterization of the gross composition of CBSs, the total polyphenol content and radical scavenging of extracts obtained from both raw and defatted cocoa bean shells were investigated. Quantification of the main polyphenolic compounds was then performed by RP-HPLC-DAD, identifying flavonoids and phenolic acids, as well as clovamide. The application of MAE and UAE resulted in a similar or superior extraction of polyphenols when compared with traditional methods; the concentration of individual polyphenols was variously influenced by the extraction methods employed. Combining MAE and UAE at 90 °C yielded the highest antiradical activity of the extract. Spectrophotometric analysis confirmed the presence of high-molecular-weight melanoidins, which were present in higher concentrations in the extracts obtained using MAE and UAE, especially starting from raw material. In conclusion, these results emphasize the efficiency of MAE and UAE techniques in obtaining polyphenol-rich extracts from CBS and confirm this cocoa by-product as a valuable biomass for the recovery of antioxidant compounds, with a view to possible industrial scale-up.

Impact of traditional and innovative cooking techniques on Italian black rice (Oryza sativa L., Artemide cv) composition.

Due to its high polyphenol content, black rice plays a significant role in good nutrition; however, these antioxidant compounds are affected by heat treatments required for the rice consumption. The aim of this work was to investigate how cooking affects the composition of Artemide black rice, comparing innovative methods, such as sous vide, with traditional domestic techniques (risotto and pilaf). Proteins and ashes were not affected by cooking, except for pilaf rice, where a 42 % ashes decrease was observed; fiber content increased after all cooking methods, reaching a 29 % increase in the risotto. Antioxidant activity, total polyphenols, anthocyanins and proanthocyanidins were reduced on average of 40 %, 34 %, 43 % and 39 %, respectively. Individual anthocyanins decreased, while phenolic acids and other flavonoids presented different behaviours, also depending if considered in their free or bound form. Cyanidin-3-O-glucoside was reduced up to 56 % in the sous vide cooked rice at 99 °C, and only by 45 % and 37 % in the risotto and sous vide cooked rice at 89 °C, respectively. Traditional risotto preparation and the innovative sous vide cooking at 89 °C also maintained the highest antioxidant polyphenols content, saving 63 % of the antioxidant activity in respect to the raw black rice. Concluding, these last techniques can be suggested for a better preservation of bioactive compounds.

Microencapsulation of a Pickering Oil/Water Emulsion Loaded with Vitamin D3.

The ionotropic gelation technique was chosen to produce vitamin D3-loaded microparticles starting from oil-in-water (O/W) Pickering emulsion stabilized by flaxseed flour: the hydrophobic phase was a solution of vitamin D3 in a blend of vegetable oils (ω6:ω3, 4:1) composed of extra virgin olive oil (90%) and hemp oil (10%); the hydrophilic phase was a sodium alginate aqueous solution. The most adequate emulsion was selected carrying out a preliminary study on five placebo formulations which differed in the qualitative and quantitative polymeric composition (concentration and type of alginate selected). Vitamin D3-loaded microparticles in the dried state had a particle size of about 1 mm, 6% of residual water content and excellent flowability thanks to their rounded shape and smooth surface. The polymeric structure of microparticles demonstrated to preserve the vegetable oil blend from oxidation and the integrity of vitamin D3, confirming this product as an innovative ingredient for pharmaceutical and food/nutraceutical purposes.

The Impact of Different Types of Rice and Cooking on Postprandial Glycemic Trends in Children with Type 1 Diabetes with or without Celiac Disease.

The aims of this study were to evaluate: (i) the chemical and nutritional composition of rice before and after cooking and (ii) postprandial glycemic impacts in children and adolescents with type 1 diabetes (T1D) after eating two different types of rice ("Gigante Vercelli" white rice and "Artemide" black rice) or white rice cooked "risotto" style or boiled using an advanced hybrid closed loop (AHCL) system (Tandem Control-IQTM). General composition and spectrophotometric analyses of raw and cooked rice were performed. Eight T1D subjects (four males and four females, aged 11 ± 1.4 years), two with celiac disease (CD), using an AHCL system were enrolled. "Gigante Vercelli" white rice cooked as risotto or boiled and boiled "Artemide" rice were prepared by the same cook on two evenings. Continuous glucose monitoring metrics were evaluated for 12 h after meal consumption. Total dietary fiber was higher for both rice types after cooking compared with raw rice. Cooking as risotto increased polyphenols and antioxidants (p < 0.05) in both rice varieties, and total starch decreased after boiling (p < 0.05) in white rice. There was a significant peak in glycemia after consuming risotto and boiled white rice (p < 0.05), while the mean glycemic peak remained <180 mg/dL in individuals eating boiled Artemide rice. There were no significant differences in automatic basal or auto-bolus insulin deliveries by the AHCL according to different types of rice or cooking method. Our findings suggest that glycemic trends are impacted by the different chemical and nutritional profiles of rice but are nevertheless well controlled by AHCL systems.

Impact of Japanese beetles (Popillia japonica Newman) on the chemical composition of two grape varieties (Nebbiolo and Erbaluce) grown in Italy.

The effect of Japanese beetles (P. japonica) on critical quality indicators of Nebbiolo and Erbaluce grapes, specifically on their phenolic and volatile composition, was assessed. Adult beetle symptoms include extended skeletonization of leaves. Leaves are frequently left with their mid-vein intact but, when severely damaged, quickly turn brown. However, the plant tends to recover by generating a new leaf apparatus and the grapes reach ripeness. It emerged that the phenolic content of grapes produced by plants attacked by P. japonica (396 and 550 mg/kg, Nebbiolo and Erbaluce respectively) was generally higher when compared to healthy plants (266 and 188 mg/kg, Nebbiolo and Erbaluce respectively). Similarly, in the (red) Nebbiolo cultivar, the anthocyanin content was significantly lower in grapes produced with healthy plants. The influence of P. japonica on the volatile composition of Nebbiolo and Erbaluce grapes showed a total volatile fraction of affected grapes (433 and 439 µg/kg, respectively) significantly higher than the one related to healthy grapes (391 and 386 µg/kg, respectively). In response to the attack by P. japonica the plant significantly increases the content of some volatile compounds such as hexanal, (E)-2-hexenal, 1-hexanol, (E)-2-hexen-1-ol and phenyl ethyl alcohol.

Tracing the Volatilomic Fingerprint of the Most Popular Italian Fortified Wines.

The aim of the current study was to provide a useful platform to identify characteristic molecular markers related to the authenticity of Italian fortified wines. For this purpose, the volatilomic fingerprint of the most popular Italian fortified wines was established using headspace solid-phase microextraction combined with gas chromatography-mass spectrometry (HS-SPME/GC-MS). Several volatile organic compounds (VOCs), belonging with distinct chemical groups, were identified, ten of which are common to all the analyzed fortified Italian wines. Terpenoids were the most abundant chemical group in Campari bitter wines due to limonene's high contribution to the total volatilomic fingerprint, whereas for Marsala wines, alcohols and esters were the most predominant chemical groups. The fortified Italian wines VOCs network demonstrated that the furanic compounds 2-furfural, ethyl furoate, and 5-methyl-2-furfural, constitute potential molecular markers of Marsala wines, while the terpenoids nerol, α-terpeniol, limonene, and menthone isomers, are characteristic of Vermouth wines. In addition, butanediol was detected only in Barolo wines, and ß-phellandrene and ß-myrcene only in Campari wines. The obtained data reveal an adequate tool to establish the authenticity and genuineness of Italian fortified wines, and at the same time constitute a valuable contribution to identify potential cases of fraud or adulteration to which they are subject, due to the high commercial value associated with these wines. In addition, they contribute to the deepening of scientific knowledge that supports its valorization and guarantee of quality and safety for consumers.

Screening of Different Essential Oils Based on Their Physicochemical and Microbiological Properties to Preserve Red Fruits and Improve Their Shelf Life.

Strawberries and raspberries are susceptible to physiological and biological damage. Due to the consumer concern about using pesticides to control fruit rot, recent attention has been drawn to essential oils. Microbiological activity evaluations of different concentrations of tested EOs (cinnamon, clove, bergamot, rosemary and lemon; 10% DMSO-PBS solution was used as a diluent) against fruit rot fungal strains and a fruit-born human pathogen (Escherichia coli) indicated that the highest inhibition halos was found for pure cinnamon and clove oils; according to GC-MS analysis, these activities were due to the high level of the bioactive compounds cinnamaldehyde (54.5%) in cinnamon oil and eugenol (83%) in clove oil. Moreover, thermogravimetric evaluation showed they were thermally stable, with temperature peak of 232.0 °C for cinnamon and 200.6/234.9 °C for clove oils. Antibacterial activity evaluations of all tested EOs at concentrations from 5-50% (v/v) revealed a concentration of 10% (v/v) to be the minimum inhibitory concentration and minimum bactericidal concentration. The physicochemical analysis of fruits in an in vivo assay indicated that used filter papers doped with 10% (v/v) of cinnamon oil (stuck into the lids of plastic containers) were able to increase the total polyphenols and antioxidant activity in strawberries after four days, with it being easier to preserve strawberries than raspberries.

A Rapid and Efficient Loop-mediated Isothermal Amplification (LAMP) Assay for the Authentication of Food Supplements Based on Maitake (Grifola Frondosa).

Grifola frondosa ("Maitake") is an edible fungus with several nutraceutical properties, largely used in traditional medicine. The increased use of Maitake as a food supplements ingredient raised the need of accurate authentication methods since the morphological identification of G. frondosa is not feasible in formulated food supplements. We developed a diagnostic tool based on loop-mediated isothermal AMPlification (LAMP) for the detection of G. frondosa in food supplements. First, a modified CTAB protocol for DNA extraction from food supplements has been set up and it has been shown to be able to isolate amplifiable total genomic material from different types of commercial products. Subsequently, the LAMP assay confirmed high specificity and good analytical sensitivity, allowing to detect up to 0.62 pg of genomic DNA in less than 20 min. Ten related fungal species resulted negative, confirming the specificity of the assay. The presence of Maitake in commercial food supplements was confirmed, except for one, revealing a mislabeling (or a food fraud). This assay proved to be a rapid powerful tool for food authentication purposes and routine inspections at any level of the supply chain of Maitake-based products and it can be used as a model for other quality control assays of fungal food products. Supplementary Information: The online version contains supplementary material available at 10.1007/s12161-022-02235-0.

ADI pathway and histidine decarboxylation are reciprocally regulated in Lactobacillus hilgardii ISE 5211: proteomic evidence.

Amine production by amino acid decarboxylation is a common feature that is used by lactic acid bacteria (LAB) to complement lactic fermentation, since it is coupled with a proton-extruding antiport system which leads to both metabolic energy production and the attenuation of intracellular acidity. Analogous roles are played in LAB by both malolactic fermentation (MLF) and the arginine deiminase (ADI) pathway. The present investigation was aimed at establishing reciprocal interactions between amino acid decarboxylation and the two above mentioned routes. The analyses were carried out on a Lactobacillus hilgardii strain (ISE 5211) that is able to decarboxylate histidine to histamine, which had previously been isolated from wine and whose complete genome is still unknown. The 2DE proteomic approach, followed by MALDI TOF-TOF and De Novo Sequencing, was used to study the protein expression levels. The experimental evidence has indicated that malate does not influence histidine decarboxylase (HDC) biosynthesis and that histidine does not affect the malolactic enzyme level. However, the expression of the ADI route enzymes, arginine deiminase and ornithine transcarbamylase, is down-regulated by histidine: this biosynthetic repression is more important (4-fold) in cultures that are not supplemented with arginine, but is also significant (2-fold) in an arginine supplemented medium that normally induces the ADI pathway. On the other hand, arginine partially represses HDC expression, but only when histidine and arginine are both present in the culture medium. This proteomic study has also pointed out a down-regulation exerted by histidine over sugar metabolism enzymes and a GroEL stress protein. These data, together with the reciprocal antagonism between arginine deimination and histidine decarboxylation, offer clue keys to the understanding of the accumulation of lactate, amine, ammonia and ethylcarbamate in wine, with consequent implications on different health risk controls.

Cooking of Artemide Black Rice: Impact on Proximate Composition and Phenolic Compounds.

The consumption of black rice has grown in recent years due to its particular organoleptic properties and high content of antioxidant polyphenols, which make it a sort of natural functional food. However, heat treatment applied during cooking can influence the content and the composition of antioxidant components, particularly anthocyanins, the main compounds of black rice, responsible for its color. The aim of this work was to evaluate the impact of different cooking techniques (boiling, microwaves oven, under pressure pot and risotto preparation) on the chemical and nutritional composition of the Italian Artemide black rice. Different cooking methods had significant and different impact on rice composition. Proximate composition was not affected by cooking, except for moisture, which increased, and fiber content, which decreased. Total polyphenols, total anthocyanin content, and antioxidant capacity were reduced; moreover, anthocyanins and phenolic acids determined by HPLC-DAD generally decreased, with the only exception of protocatechuic acid. The risotto preparation was the most useful cooking technique to preserve anthocyanins and antioxidant activity. Our results demonstrated the importance to study cooking methods and to evaluate their impact on rice characteristics, in order to preserve its nutritional and beneficial properties.

Gentianose: Purification and structural determination of an unknown oligosaccharide in grape seeds.

Grape seeds are among the main constituents of grape pomace, ranging between 20% and 30% of the wet matrix; however, their oligosaccharide composition has not been studied. This paper describes the purification and the identification of low molecular weight oligosaccharides contained in an EtOH/water extract of grape seeds. A sequential two-step purification by size exclusion chromatography was carried out to fractionate compounds according to molecular weights. Chemical characterization of the combined fractions was performed by Magnetic Resonance Spectroscopy and Gas Chromatography-Mass Spectrometry analyses. The separation process gave two fractions abundant in sucrose and glucose. A third fraction containing trisaccharides was acetylated allowing the purification of the main trisaccharide. The structure elucidation of the acetylated product made it possible to identify gentianose, a predominant carbohydrate reserve found in the storage roots of perennial Gentiana lutea. Grape seeds are wine industry by-products and the obtained results suggest the importance of their recovery.

Permeate from cheese whey ultrafiltration is a source of milk oligosaccharides.

Previously undescribed oligosaccharides in bovine cheese whey permeate were characterized by a combination of nanoelectrospray Fourier Transform Ion Cyclotron Resonance (nESI-FTICR) mass spectrometry and matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance (MALDI-FTICR) mass spectrometry. Oligosaccharide composition was elucidated by collision-induced dissociation within the ICR cell. In addition to sialyllactose (the most abundant oligosaccharide in bovine colostrum), we identified 14 other oligosaccharides, half of which have the same composition of human milk oligosaccharides. These oligosaccharides could potentially be used as additives in infant formula and products for the pharmaceutical industry. Because whey permeate is a by-product from the production of whey protein concentrate (WPC) and is readily available, it is an attractive source of oligosaccharides for potential application in human nutrition.

Distribution of bioactive compounds in pearled fractions of tritordeum.

Hexaploid tritordeum is the amphidiploid cereal derived from the cross between wild barley and durum wheat. The present study compares two cultivars of tritordeum with other cereals grown in the same experimental area to weigh up its potential use as ingredient for health-valued foods. Tritordeum shows 2.5-fold higher concentration of lutein than common wheat and barley, and 1.2-fold higher than durum wheat, while the concentration of ß-glucans is 5 folds lower than the one observed for barley. Based on the distribution of bioactive compounds in pearled fractions, the use of whole-grain flours seems the best way to exploit the antioxidant potential of tritordeum. Nevertheless, the internal layers of the kernel of this cereal are characterized on average by high concentrations of antioxidants (32.0 mg/kg and 518 mg/kg soluble and cell wall-bound phenolic acids, respectively), making tritordeum interesting also for the production of refined flours rich in bioactive compounds.

The impact of distillation process on the chemical composition and potential prebiotic activity of different oligosaccharidic fractions extracted from grape seeds.

The study was designed to evaluate how the distillation process is able to affect the composition and bioactivity of oligosaccharides contained in the grape seeds. Different oligosaccharidic fractions have been extracted both before and after grape pomace distillation in order to valorize this by-product. A multistep solid-phase extraction approach (C-18 and carbograph cartridges) has been applied to purify and fractionate the oligosaccharidic compounds. Chemical characterization of the fractions was performed using a UPLC-ESI-MSn method. Complex oligosaccharides consist principally of neutral oligosaccharides rich in arabinose and glucose. Then, the oligosaccharides contained in the different fractions as potential functional ingredients with prebiotic activity toward well-known probiotic bacteria, such as Lactobacillus acidophilus and Lactobacillus plantarum, were evaluated. Data showed how, in some combination, oligosaccharidic fractions obtained may be considered a novel "functional ingredient" with potential prebiotic activity mainly towards L. acidophilus.

Cocoa hulls polyphenols stabilized by microencapsulation as functional ingredient for bakery applications.

Cocoa hulls are a potential source of polyphenols to be used as "functional ingredients" in foods, but their low stability to oxidation and thermal degradation limits their practical application. The aim of this study was to microencapsulate cocoa hulls phenolic extracts through spray-drying, in order to produce new heat stable ingredients for bakery products. Polyphenols were extracted using water and ethanol under different conditions. The best performing extract (water/ethanol 50:50), containing 93.3 mg of total polyphenols per gram of dry extract, was spray-dried with and without stabilizing agents (maltodextrins and/or gum Arabic), obtaining seven different powders. These were first tested for their stability, showing a total phenolic content and an antioxidant activity stable up to 90 days. The powders were then used to evaluate their baking stability in a model biscuit; the microencapsulation using an 80:20 ratio of maltodextrins to the dry extract allowed obtaining the most stable powder, with a total polyphenol content unaffected by the baking process.

Characterization of peel and pulp proanthocyanidins and carotenoids during ripening in persimmon "Kaki Tipo" cv, cultivated in Italy.

The proanthocyanidins and carotenoids composition of peel and pulp from persimmon "Seedless Pollination Variant Non-Astringent" cultivar during ripening was evaluated. Phloroglucinolysis method, combined with HPLC separation, confirmed three terminal units, the flavonol myricetin, in addition to the typical flavan-3-ols catechin and epicatechin. The extender units were epicatechin, epigallocatechin, epigallocatechin-3-O-gallate, epicatechin-3-O-gallate, and catechin. Proanthocyanidins were characterized by a considerable level of 3-O-galloylation (34-60%), showing a high prodelphinidin content (29-46%) and a mean degree of polymerization between 55 and 10. A carotenoid increase was observed during fruit ripening both in the peel and the pulp, with the highest contents respectively, for the peel and the pulp of 42.11 mg/Kg and 13.32 mg/Kg, determined at the almost fully mature stage.

Detection of biogenic amine producer bacteria in a typical Italian goat cheese.

The aim of this study was to research decarboxylating bacterial strains and biogenic amine content in a typical Italian goat cheese (Robiola di Roccaverano). The study was performed on fresh and ripened samples of goat cheese manufactured from industrial and artisanal producers. Sixty-seven bacterial strains isolated showed decarboxylating activity, and Enterococcus faecalis was the most widespread decarboxylating species in all artisanal and industrial products. Pediococcus acidilactici and Enterococcus malodoratus were also identified as biogenic amine producers in Robiola di Roccaverano cheese. All the E. faecalis strains isolated in this study were able to decarboxylate tyrosine. Tyramine was the most abundant biogenic amine in cheese samples, while histamine was the most widespread. High amounts of these two biogenic amines were found in ripened samples (up to 2,067 mg/kg for tyramine and 1,786 mg/kg for histamine), whereas 2-phenylethylamine and tryptamine were present in almost all ripened cheeses at low concentrations. The detection of strains producing biogenic amines and the high concentrations of tyramine and histamine found in ripened Robiola di Roccaverano could represent a potential risk to the consumer.

Spray-dried polyphenolic extract from Italian black rice (Oryza sativa L., var. Artemide) as new ingredient for bakery products.

Artemide is one of the Italian pigmented rice varieties richest in polyphenols, especially anthocyanins. The aim of this work was to obtain anthocyanin-rich powders from Artemide cv, useful as functional ingredients for bakery foods. The hydroalcoholic extract prepared from Artemide black rice was processed by both spray-drying (with and without coating agents: maltodextrins: MD; arabic gum: GA) and freeze drying, in order to obtain ingredients more stable during storage and baking. The polyphenols spray-dried with MD and GA (50:50, w/w) resulted the ingredient most stable to the storage and partially protected from thermal damage during the baking in a model biscuit. The enriched biscuits showed a significantly higher content of polyphenols, antioxidant capacity and anthocyanins respect to a control biscuit. The polyphenolic extract obtained from Artemide black rice can be considered a valuable source of polyphenols to produce functional foods or microencapsulated ingredients for nutraceutical applications.

Bioactive compound and antioxidant activity distribution in roller-milled and pearled fractions of conventional and pigmented wheat varieties.

In this study, the chemical composition of pigmented wheats (yellow, purple and blue types), and the distribution of the bioactive compounds in their roller-milled and pearled fractions, were compared with conventional wheats (red and white types). Roller-milling promoted the recovery of total dietary fiber, ß-glucans, phenolic acids and anthocyanins in the bran fraction, which resulted also in a higher total antioxidant activity than the refined flour. Conversely, lutein resulted mainly concentrated in the refined flour. In the same way, the distribution pattern in the pearled fractions differ depending on the bioactive considered. The study highlights that a careful selection of the most appropriate fractionation process should be performed to produce flours rich in bioactive compounds. Roller-milling resulted useful for the production of refined flours rich in xanthophylls, with particular emphasis to the yellow-grained wheats. Contrarily, pearling could be more useful in the valorization of the health potential of anthocyanin-pigmented varieties.