Microencapsulated bitter compounds (from Gentiana lutea) reduce daily energy intakes in humans.

Mounting evidence showed that bitter-tasting compounds modulate eating behaviour through bitter taste receptors in the gastrointestinal tract. This study aimed at evaluating the influence of microencapsulated bitter compounds on human appetite and energy intakes. A microencapsulated bitter ingredient (EBI) with a core of bitter Gentiana lutea root extract and a coating of ethylcellulose-stearate was developed and included in a vanilla microencapsulated bitter ingredient-enriched pudding (EBIP). The coating masked bitterness in the mouth, allowing the release of bitter secoiridoids in the gastrointestinal tract. A cross-over randomised study was performed: twenty healthy subjects consumed at breakfast EBIP (providing 100 mg of secoiridoids) or the control pudding (CP) on two different occasions. Blood samples, glycaemia and appetite ratings were collected at baseline and 30, 60, 120 and 180 min after breakfast. Gastrointestinal peptides, endocannabinoids (EC) and N-acylethanolamines (NAE) were measured in plasma samples. Energy intakes were measured at an ad libitum lunch 3 h after breakfast and over the rest of the day (post lunch) through food diaries. No significant difference in postprandial plasma responses of gastrointestinal hormones, glucose, EC and NAE and of appetite between EBIP and CP was found. However, a trend for a higher response of glucagon-like peptide-1 after EBIP than after CP was observed. EBIP determined a significant 30 % lower energy intake over the post-lunch period compared with CP. These findings were consistent with the tailored release of bitter-tasting compounds from EBIP along the gastrointestinal tract. This study demonstrated that microencapsulated bitter secoiridoids were effective in reducing daily energy intake in humans.

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.

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.

Behind the Scenes of Anthocyanins-From the Health Benefits to Potential Applications in Food, Pharmaceutical and Cosmetic Fields.

Anthocyanins are widespread and biologically active water-soluble phenolic pigments responsible for a wide range of vivid colours, from red (acidic conditions) to purplish blue (basic conditions), present in fruits, vegetables, and coloured grains. The pigments' stability and colours are influenced mainly by pH but also by structure, temperature, and light. The colour-stabilizing mechanisms of plants are determined by inter- and intramolecular co-pigmentation and metal complexation, driven by van der Waals, π-π stacking, hydrogen bonding, and metal-ligand interactions. This group of flavonoids is well-known to have potent anti-inflammatory and antioxidant effects, which explains the biological effects associated with them. Therefore, this review provides an overview of the role of anthocyanins as natural colorants, showing they are less harmful than conventional colorants, with several technological potential applications in different industrial fields, namely in the textile and food industries, as well as in the development of photosensitizers for dye-sensitized solar cells, as new photosensitizers in photodynamic therapy, pharmaceuticals, and in the cosmetic industry, mainly on the formulation of skin care formulations, sunscreen filters, nail colorants, skin & hair cleansing products, amongst others. In addition, we will unveil some of the latest studies about the health benefits of anthocyanins, mainly focusing on the protection against the most prevalent human diseases mediated by oxidative stress, namely cardiovascular and neurodegenerative diseases, cancer, and diabetes. The contribution of anthocyanins to visual health is also very relevant and will be briefly explored.

Optimization of an Untargeted DART-HRMS Method Envisaging Identification of Potential Markers for Saffron Authenticity Assessment.

Saffron is one of the most expensive agricultural products in the world and as such, the most commonly adulterated spice, with undeclared plant-based surrogates or synthetic components simulating color and morphology. Currently, saffron quality is certificated in the international trade market according to specific ISO guidelines, which test aroma, flavor, and color strength. However, it has been demonstrated that specific adulterants such as safflower, marigold, or turmeric up to 20% (w/w) cannot be detected under the prescribed approach; therefore, there is still a need for advanced and sensitive screening methods to cope with this open issue. The current investigation aims to develop a rapid and sensitive untargeted method based on an ambient mass spectrometry ionization source (DART) and an Orbitrap™high-resolution mass analyzer to discriminate pure and adulterated saffron samples with either safflower or turmeric. The metabolic profiles of pure and adulterated model samples prepared at different inclusion levels were acquired. Unsupervised multivariate analysis was carried out based on hierarchical cluster analysis and principal component analysis as first confirmation of the discriminating potential of the metabolic profile acquired under optimized DART-HRMS conditions. In addition, a preliminary selection of potential markers for saffron authenticity was accomplished, identifying compounds able to discriminate the type of adulteration down to a concentration level of 5%.

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.

Multi-target detection of egg-white and pig gelatin fining agents in Nebbiolo-based aged red wine by means of nanoHPLC-HRMS.

The presence of residues from fining agents in wines may represent a risk for allergic consumers and a source of discomfort for others, such as vegans. Even though ELISA is the official detection method for such residues, this technique may be hindered by cross-reactivity issues, or by matrix-molecule interference due to a high polyphenol content, especially in red wines. An HRMS-based method has been developed to detect pig gelatin and egg white in experimental five-year aged Nebbiolo-based red wine. Biomarker peptides were selected, after tryptic digestion, and quantified by multitarget nanoHPLC-HRMS analysis. The method resulted in an LLOQs of 5 µg/mL in the experimental wine, and between 1 and 2 µg/mL in the buffer. This method allowed both gelatin and egg white proteins to be detected and quantified in aged red wine, while whereas the commercial ELISA kit was instead unable to detect egg white in the same samples.

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.

Molecular insight into IgE-mediated reactions to sesame (Sesamum indicum L.) seed proteins.

BACKGROUND: Food allergy is becoming a major public health concern in recent times. Several sesame seed allergenic proteins have been identified. However, sensitization toward these proteins does not follow a common and unique pattern of clinical reactivity, as shown by the differential geographic recognition of single proteins. OBJECTIVE: To evaluate the sensitization profiles of 18 Italian individuals who experienced clinical symptoms after sesame seed consumption, including 4 anaphylactic reactions. METHODS: Using an in vitro approach, we adopted a 2-dimensional electrophoretic technique combined with immunoblotting analyses by using sera from 18 Italian sesame-allergic patients. RESULTS: We showed the prevalent and almost exclusive reactivity of the sesame 11S globulin. We shed light on the active role of the basic subunit of this globulin family. The limited accessibility of this polypeptide chain, unless the interchain disulphide bonds are cleaved, may be one of the reasons for its structural/functional stability and, thus, great potential for induction of IgE reactivity. CONCLUSIONS: These results confirmed previous findings on the reactivity of the basic subunit of 11S globulin in various legume species. Moreover, this experimental approach proved to be useful for the noninvasive screening of specific reactivities in sensitized patients.

Geographical Origin Discrimination of Monofloral Honeys by Direct Analysis in Real Time Ionization-High Resolution Mass Spectrometry (DART-HRMS).

An untargeted method using direct analysis in real time and high resolution mass spectrometry (DART-HRMS) combined to multivariate statistical analysis was developed for the discrimination of two monofloral (chestnut and acacia) honeys for their geographical origins-i.e., Italy and Portugal for chestnut honey and Italy and China for acacia honey. Principal Component Analysis, used as an unsupervised approach, showed samples of clusterization for chestnut honey samples, while overlapping regions were observed for acacia honeys. Three supervised statistical approaches, such as Principal Components-Linear Discriminant Analysis, Partial Least Squares-Discriminant Analysis and k-nearest neighbors, were tested on the dataset gathered and relevant performances were compared. All tested statistical approaches provided comparable prediction abilities in cross-validation and external validation with mean values falling between 89.2-98.4% for chestnut and between 85.8-95.0% for acacia honey. The results obtained herein indicate the feasibility of the DART-HRMS approach in combination with chemometrics for the rapid authentication of honey's geographical origin.

Machine Learning Approaches Applied to GC-FID Fatty Acid Profiles to Discriminate Wild from Farmed Salmon.

In the last decade, there has been an increasing demand for wild-captured fish, which attains higher prices compared to farmed species, thus being prone to mislabeling practices. In this work, fatty acid composition coupled to advanced chemometrics was used to discriminate wild from farmed salmon. The lipids extracted from salmon muscles of different production methods and origins (26 wild from Canada, 25 farmed from Canada, 24 farmed from Chile and 25 farmed from Norway) were analyzed by gas chromatography with flame ionization detector (GC-FID). All the tested chemometric approaches, namely principal components analysis (PCA), t-distributed stochastic neighbor embedding (t-SNE) and seven machine learning classifiers, namely k-nearest neighbors (kNN), decision tree, support vector machine (SVM), random forest, artificial neural networks (ANN), naïve Bayes and AdaBoost, allowed for differentiation between farmed and wild salmons using the 17 features obtained from chemical analysis. PCA did not allow clear distinguishing between salmon geographical origin since farmed samples from Canada and Chile overlapped. Nevertheless, using the 17 features in the models, six out of the seven tested machine learning classifiers allowed a classification accuracy of ≥99%, with ANN, naïve Bayes, random forest, SVM and kNN presenting 100% accuracy on the test dataset. The classification models were also assayed using only the best features selected by a reduction algorithm and the best input features mapped by t-SNE. The classifier kNN provided the best discrimination results because it correctly classified all samples according to production method and origin, ultimately using only the three most important features (16:0, 18:2n6c and 20:3n3 + 20:4n6). In general, the classifiers presented good generalization with the herein proposed approach being simple and presenting the advantage of requiring only common equipment existing in most labs.

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.

In depth study of phenolic profile and PTP-1B inhibitory power of cold-pressed grape seed oils of different varieties.

This paper investigates the phenolic composition of 17 monocultivar commercial cold-pressed grape seed oils. Chromatographic profiles showed the presence of more than 28 molecules, 11 of which were successfully identified by HPLC-DAD-MS-TOF and HPLC-FLD analysis. Pinoresinol, ethyl caffeate and ethyl gallate were detected for the first time in these oils. The total phenolic content ranged between 0.83 mg/kg for Viognier sample to 15.16 mg/kg for Merlot org sample. The detected ethyl esters can be suggested as markers to evaluate the intensity of fermentation in grape seeds before oil extraction, and to control the sensorial quality of the produced oils. In addition, the inhibitory power of these phenolic extracts against Protein Tyrosine Phosphatase 1B enzyme (PTP-1B), overexpressed in type-two diabetes, was investigated for the first time. Data highlighted a good correlation between total phenolic content and inhibitory power, with pinoresinol, p-coumaric acid and quercetin making the greater contributions.

Mass Spectrometry-Based Untargeted Proteomics for the Assessment of Food Authenticity: The Case of Farmed Versus Wild-Type Salmon.

Background: Omics technologies have been widely applied in different fields, among which, proteomics is gaining increasing interest for its application to the authenticity of food products. MS, typically coupled with LC, represents a key technique for proteomics-related studies dedicated to fish and other seafood products by using a bottom-up approach. Objective and Methods: In this paper, the optimization of an untargeted proteomics-based method using LC separation and MS detection relying on a quadrupole time-of-flight mass spectrometer is described and applied to the analysis of Canadian farmed and wild-type salmon, followed by statistical analysis based on principal component (PC) analysis. Results and Conclusions: This untargeted approach, using a data-independent acquisition MS scheme, demonstrated the ability to effectively discriminate salmon belonging to the two classes. Furthermore, selected peptides showing high loadings on PC1 could represent potential candidate peptide markers able to discriminate farmed from wild-type salmon samples in the future.

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.

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.

Assessing fish authenticity by direct analysis in real time-high resolution mass spectrometry and multivariate analysis: discrimination between wild-type and farmed salmon.

The constant increase in seafood consumption worldwide has led to a parallel growth of the incidence of products obtained by aquaculture on the market, but also of the fraudulent commercialization of farmed products as wild-type ones. A careful characterization of the lipid component of seafood products based on chromatography-mass spectrometry techniques has been reported as a promising approach to reliably differentiate farmed from wild-type products. In this context, a fast method based on Direct Analysis in Real Time (DART) coupled to High Resolution Mass Spectrometry (HRMS) based on a single stage Orbitrap mass analyzer, integrated by Principal Component Analysis (PCA), was developed in the present study and applied to scout for spectral features useful to discriminate wild-type from farmed salmon of Salmo salar species. In particular, normalized intensities obtained for the 30 most intense signals (all referred to fatty acids, FA) detected in negative ion DART-HRMS spectra of the lipid extracts of salmon fillets [26 wild-type from Canada, 74 farmed from Canada (25), Norway (25) and Chile (24)] were considered as the variables for PCA. The scatterplot referred to the first two principal components showed a clear distinction between wild-type and farmed salmon, which gathered as a unique cluster, despite the remarkable differences in their geographical origin. In accordance with previous studies based on more complex and time-demanding analytical approaches, three saturated (14:0, 16:0 and 18:0) FA, along with unsaturated ones having 20 or 22 carbon atoms, were found as the main discriminating variables for wild-type salmons, whereas FA with compositions 18:1, 18:2, 18:3 and several oxidized forms arising from them were found to have a significantly higher incidence in farmed salmon. The method was further validated by Discriminant Analysis (DA) performed on the same dataset used for PCA integrated by data obtained from 6 commercial samples, putatively referred to farmed Norwegian salmon. Results showed that 100% of the latter were correctly classified as farmed type. Relative abundances of DART-HRMS signals related to specific FA appear then very promising for the differentiation of wild-type salmon from farmed ones, a very relevant issue in the context of consumers' protection from seafood frauds.

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.

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.