Authentication of Laying Hen Housing Systems Based on Egg Yolk Using 1H NMR Spectroscopy and Machine Learning.

(1) Background: The authenticity of eggs in relation to the housing system of laying hens is susceptible to food fraud due to the potential for egg mislabeling. (2) Methods: A total of 4188 egg yolks, obtained from four different breeds of laying hens housed in colony cage, barn, free-range, and organic systems, were analyzed using 1H NMR spectroscopy. The data of the resulting 1H NMR spectra were used for different machine learning methods to build classification models for the four housing systems. (3) Results: The comparison of the seven computed models showed that the support vector machine (SVM) model gave the best results with a cross-validation accuracy of 98.5%. The test of classification models with eggs from supermarkets showed that only a maximum of 62.8% of samples were classified according to the housing system labeled on the eggs. (4) Conclusion: The classification models developed in this study included the largest sample size compared to the literature. The SVM model is most suitable for evaluating 1H NMR data in terms of the hen housing system. The test with supermarket samples showed that more authentic samples to analyze influencing factors such as breed, feeding, and housing changes are required.

Sustainable In Silico-Supported Ultrasonic-Assisted Extraction of Oligomeric Stilbenoids from Grapevine Roots Using Natural Deep Eutectic Solvents (NADES) and Stability Study of Potential Ready-to-Use Extracts.

Grapevine roots, as a side-stream of a vineyard, are a sustainable resource for the recovery of oligomeric stilbenoids, such as the bioactive r-viniferin. The aim of this study is to evaluate an in silico-supported method, based on the Conductor-like Screening Model for Real Solvents (COSMO-RS), for selection of environmentally friendly natural deep eutectic solvents (NADES) with regard to the extraction of grapevine roots. The most suitable NADES system for ultrasonic-assisted extraction of r-viniferin was choline chloride/1,2-propanediol. The optimal extraction parameters for r-viniferin were determined using single-factor experiments as follows: choline chloride/1,2-propanediol 1/2 mol/mol, 10 wt% H2O, biomass/NADES ratio 1/10 g/g, and 10 min extraction time. Under optimized conditions, the extraction yield of r-viniferin from grapevine roots reached 76% of the total r-viniferin content. Regarding stability, stilbenoids in choline chloride/1,2-propanediol remained stable during 128 days of storage at ambient temperature. However, fructose/lactic acid-based NADES were observed to degrade stilbenoids; therefore, the removal of the NADES will be of interest, with a suitable method implemented using Amberlite® XAD-16N resin. As green solvents, the NADES have been used as effective and environmentally friendly extractants of stilbenoid-containing extracts from grapevine roots for potential applications in the cosmetic and pharmaceutical industry or as nutraceuticals in the food industry.

Formation of amyloid fibrils from ovalbumin under Ohmic heating.

Ohmic heating (OH) is an alternative sustainable heating technology that has demonstrated its potential to modify protein structures and aggregates. Furthermore, certain protein aggregates, namely amyloid fibrils (AF), are associated with an enhanced protein functionality, such as gelation. This study evaluates how Ohmic heating (OH) influences the formation of AF structures from ovalbumin source under two electric field strength levels, 8.5 to 10.5 and 24.0-31.0 V/cm, respectively. Hence, AF aggregate formation was assessed over holding times ranging from 30 to 1200 sunder various environmental conditions (3.45 and 67.95 mM NaCl, 80, 85 and 90 °C, pH = 7). AF were formed under all conditions. SDS-PAGE revealed that OH had a higher tendency to preserve native ovalbumin molecules. Furthermore, Congo Red and Thioflavin T stainings indicated that OH reduces the amount of AF structures. This finding was supported by FTIR measurements, which showed OH samples to contain lower amounts of beta-sheets. Field flow fractioning revealed smaller-sized aggregates or aggregate clusters occurred after OH treatment. In contrast, prolonged holding time or higher treatment temperatures increased ThT fluorescence, beta-sheet structures and aggregate as well as cluster sizes. Ionic strength was found to dominate the effects of electric field strength under different environmental conditions.

In Silico-Assisted Isolation of trans-Resveratrol and trans-ε-Viniferin from Grapevine Canes and Their Sustainable Extraction Using Natural Deep Eutectic Solvents (NADES).

Grapevine canes are an important source of bioactive compounds, such as stilbenoids. This study aimed to evaluate an in silico method, based on the Conductor-like Screening Model for Real Solvents (COSMO-RS) to isolate stilbenoids from a grapevine cane extract by offline heart-cut high-performance countercurrent chromatography (HPCCC). For the following extraction of resveratrol and ε-viniferin from grapevine canes, natural deep eutectic solvents (NADES) were used as an environmentally friendly alternative to the traditionally used organic solvents. In order to evaluate a variety of combinations of hydrogen bond acceptors (HBAs) and hydrogen bond donors (HBDs) for the targeted extraction of stilbenoids, COSMO-RS was applied. In particular, ultrasonic-assisted extraction using a solvent mixture of choline chloride/1,2-propanediol leads to higher extraction yields of resveratrol and ε-viniferin. COSMO-RS calculations for NADES extraction combined with HPCCC biphasic solvent system calculations are a powerful combination for the sustainable extraction, recovery, and isolation of natural products. This in silico-supported workflow enables the reduction of preliminary experimental tests required for the extraction and isolation of natural compounds.

Metabolic and microbial analyses of the surface and inner part of wet-aged and dry-aged beef.

The effects of aging and microbial growth on the metabolome of aged beef were investigated in this study. The metabolome of beef is influenced by the aging method applied. This includes the aging-related changes in metabolism and the presence of microorganisms on the beef during aging that may affect the beef and its quality. The inner part and the trimmed surface of dry-aged (the surface of dry-aged beef is also called the "crust" due to its drying during aging) and wet-aged beef were analyzed by 1 H nuclear magnetic resonance (NMR) spectroscopy over aging periods up to 28 days at intervals of 7 days, and the former also by microbiological analysis. The metabolome detected by 1 H NMR spectroscopy demonstrated changes over the aging time of beef and differed depending on the sampling location (surface or inner part of beef). The influence of the microbiota on changes in the metabolome can be negligible due to the low microbial growth on the surface of dry-aged beef (<3 log CFU/g). Therefore, the aging-related metabolism postmortem of the analyzed dry-aged beef might be the main factor for metabolic changes. The significantly (p < 0.05) higher amino acids and inosine concentrations and lower inosine 5'-monophosphate concentrations suggested enhanced protein degradation and energy metabolism in the wet-aged beef compared to the dry-aged beef, probably due to the combined influence of the aging and the microbiota on the wet-aged beef and, thus, its metabolic changes.

Variations in the Metabolome of Unaged and Aged Beef from Black-and-White Cows and Heifers by 1H NMR Spectroscopy.

(1) Background: The selection of raw material and the postmortem processing of beef influence its quality, such as taste. In this study, the metabolome of beef from cows and heifers is examined for differences during aging. (2) Methods: Thirty strip loins from eight heifers and seven cows (breed code: 01-SBT) were cut into ten pieces and aged for 0, 7, 14, 21 and 28 days. Samples from the left strip loins were wet-aged in vacuum, while samples from right strip loins were dry-aged at 2 °C and 75% relative humidity. The beef samples were extracted with methanol-chloroform-water, and the polar fraction was used for 1H NMR analysis. (3) Results: The PCA and OPLS-DA showed that the metabolome of cows and heifers varied. Eight metabolites revealed significant differences (p < 0.05) in the samples from cows and heifers. The aging time and aging type of beef also affected the metabolome. Twenty-eight and 12 metabolites differed significantly (p < 0.05) with aging time and aging type, respectively. (4) Conclusions: The variations between cows and heifers and aging time affect the metabolome of beef. By comparison, the influence of aging type is present but less pronounced.

Metabolic, proteomic and microbial changes postmortem and during beef aging.

The purpose of this review is to provide an overview of the current knowledge about proteomic and metabolic changes in beef, the microbiological alteration postmortem and during aging, and observe the influence on beef quality parameters, such as tenderness, taste and flavor. This review will also focus on the different aging types (wet- and dry-aging), the aging or postmortem time of beef and their effect on the proteome and metabolome of beef. The Ca2+ homeostasis and adenosine 5'-triphosphate breakdown are the main reactions in the pre-rigor phase. After rigor mortis, the enzymatic degradation of connective tissues and breakdown of energy metabolism dominate molecular changes in beef. Important metabolic processes leading to the formation of saccharides, nucleotides, organic acids (e.g. lactic acid), creatine and fatty acids are considered in this context as possible flavor precursors or formers of beef flavor and taste. Flavor precursors are substrates for lipid oxidation, Strecker degradation and Maillard reaction during cooking or roasting. The findings presented should serve as a basis for a better understanding of beef aging and its molecular effects and are intended to contribute to meeting the challenges of improving beef quality.

Effect of sampling position in fresh, dry-aged and wet-aged beef from M. Longissimus dorsi of Simmental cattle analyzed by 1H NMR spectroscopy.

The aging of beef affects the metabolome and, thus, its quality, such as taste or tenderness. In addition to the aging method, intrinsic factors, such as breed, feed and muscle type, also have an effect on beef's metabolome. It is not known yet whether the position of the sampling in large muscles also has an influence on beef's metabolome and its aging outcome. The effect of the sampling position in M. longissimus dorsi as a large muscle was investigated in dry-aged and wet-aged beef over an aging period of 28 days. In this study, we analyzed 360 samples out of the entire length of M. longissimus dorsi of 18 'Simmental' young bulls by 1H NMR spectroscopy. The position in the muscle affected the polar fraction of metabolome of non-aged and aged beef significantly. However, sampling position did not overlay significant differences in the metabolome of dry-aged and wet-aged beef. The aging time of beef also had a significant effect on the metabolome. Marker metabolites, such as leucine, isoleucine, inosine 5'-monophosphate and hypoxanthine, were found to be indicative of the aging time applied. In addition, marker metabolites (lactic acid, anserine, O-acetyl-L-carnitine) were identified for the aging type applied.

Analysis of aging type- and aging time-related changes in the polar fraction of metabolome of beef by 1H NMR spectroscopy.

The tenderness and taste of beef is improved by either dry- or wet-aging or a combination of both. The objective was to develop a validated method for detecting differences in the polar fraction of metabolome in dry-aged and wet-aged beef over the aging time and quantifying the metabolites of interest by 1H NMR spectroscopy using beef. Sixty strip loin (M. longissimus dorsi) samples aged in different ways (wet-aging vs. dry-aging) and aging times (0, 7, 14, 21, 28 days) were analyzed. The aging type could be defined by linear discriminant analysis with an accuracy of 95%. Ten (lactic acid, alanine, methionine, fumaric acid, inosine, inosine monophosphate, creatine, betaine, carnosine and hypoxanthine) out of eighteen metabolites differ significantly (p < 0.05) in content depending on the aging type. Fifteen metabolites in dry-aged and ten in wet-aged beef correlate with the aging time (r > 0.7, <-0.7), which shows significant aging time-related effects on the polar fraction of metabolome.

Rapid UV/Vis Spectroscopic Dye Authentication Assay for the Determination and Classification of Reactive Dyes, Monascus Pigments, and Natural Dyes in Coloring Foodstuff.

Food authenticity in the field of food dyes can be interpreted as the correctness of the coloring ingredients indicated. The Rapid UV/vis Spectroscopic Dye Authentication Assay (RaSDAY) presented in this work was used to verify the authenticity of water-soluble reddish colorings for food use. RaSDAY includes the processing of samples under different experimental conditions with pH variations and heat exposure. The absorbances measured are analyzed by principal component analysis and a k-nearest neighbors algorithm. As a result, classification of anthocyanins, betalains, and carmine and the detection of Monascus pigments, undeclared artificial food dyes, and reactive textile azo dyes can be performed by utilizing a rapid screening method. In 17 out of 20 samples of coloring food additives that were included in this work, reactive dyes, unpermitted Monascus pigments, and artificial food dyes were detected using the developed method. "Reactive Red 120", "Reactive Red 195", and "Reactive Red 198" were identified by subsequent 1H NMR spectroscopy in eight of those samples.

Matrix- and Technology-Dependent Stability and Bioaccessibility of Strawberry Anthocyanins during Storage.

Anthocyanins are often associated with health benefits. They readily degrade during processing and storage but are also dependent on the matrix conditions. This study investigated how strawberry anthocyanins are affected by preservation technologies and a relatively protein-rich kale juice addition during storage. A strawberry-kale mix was compared to a strawberry-water mix (1:2 wt; pH 4), untreated, thermally, pulsed electric fields (PEF) and high-pressure processing (HPP) treated, and evaluated for anthocyanin stability and bioaccessibility during refrigerated storage. The degradation of strawberry anthocyanins during storage followed first-order kinetics and was dependent on the juice system, preservation technology and anthocyanin structure. Generally, the degradation rate was higher for the strawberry-kale mix compared to the strawberry-water mix. The untreated sample showed the highest degradation rate, followed by HPP, PEF and, then thermal. The relative anthocyanin bioaccessibility after gastric digestion was 10% higher for the thermally and PEF treated samples. Anthocyanin bioaccessibility after intestinal digestion was low due to instability at a neutral pH, especially for the strawberry-kale mix, and after thermal treatment. The storage period did not influence the relative bioaccessibility; yet, the absolute content of bioaccessible anthocyanins was decreased after storage. This research further presents that processing and formulation strongly affect the stability and bioaccessibility of anthocyanins during storage.

Semisynthetic preparation and isolation of dimeric procyanidins B1-B8 from roasted hazelnut skins (Corylus avellana L.) on a large scale using countercurrent chromatography.

Dimeric procyanidins B1-B8 were produced via semisynthesis from a polymeric proanthocyanidin fraction of hazelnut skins (Corylus avellana L.). This polymeric fraction was found to consist mostly of (+)-catechin and (-)-epicatechin as upper units. Therefore, according to the choice of nucleophile agent, it is possible to semisynthesize dimeric procyanidins B1, B3, B6, and B7 with (+)-catechin and B2, B4, B5, and B8 with (-)-epicatechin. The semisynthetic mixtures were separated on a preparative scale using high-speed countercurrent chromatography (HSCCC) and low-speed rotary countercurrent chromatography (LSRCCC). C4 → C8 linked dimeric procyanidins B1-B4 were isolated in amounts of 350-740 mg. To the best of the authors' knowledge this is the first study isolating dimeric procyanidins B1-B8 in large amounts with countercurrent chromatography. Moreover, the dimeric prodelphinidins B1, B2, and B3 and their structural elucidation by (1)H NMR spectroscopy without derivatization are described for hazelnuts as natural compounds for the first time.

Development of a novel adsorptive membrane chromatographic method for the fractionation of polyphenols from bilberry.

A novel membrane chromatographic method with a membrane adsorber (Sartobind S) has been developed on the laboratory scale that allows a fractionation of bilberry (Vaccinium myrtillus) constituents into the following three groups of polyphenols: anthocyanins, copigments, and polymers. By using this methodology, a pure anthocyanin fraction free of other copigments and polymeric phenols can be obtained. Using this approach, it provides fractions allowing a more thorough testing of the biological effects of the individual groups of bilberry polyphenols as well as the study of possible synergistic effects between these different groups of bioactive constituents from bilberry.