Aroma Properties of Cocoa Fruit Pulp from Different Origins.

Cocoa pulp occurs as a by-product of cocoa bean production and can be repurposed to different food applications, such as jams, fruit preparations and beverages, improving the sustainability of cocoa production, as well as the livelihoods of cocoa farmers. In this work, aroma-active compounds of fresh cocoa fruit pulps from different origins were investigated by applying aroma extract dilution analyses in combination with gas chromatography-mass spectrometry/olfactometry for identification. In total, 65 aroma-active compounds were determined in four different pulps originating from Indonesia, Vietnam, Cameroon, and Nicaragua. Vietnamese pulp showed the highest number of aroma-active regions, while Cameroonian pulp accounted for the lowest. Moreover, Cameroonian cocoa pulp showed the lowest FD factors. Overall, the odorants with the highest FD factors were trans-4,5-epoxy-(E)-decenal, 2- and 3-methylbutanoic acid, 3-(methylthio)propanal, 2-isobutyl-3-methoxypyrazine, (E,E)-2,4-nonadienal, (E,E)-2,4-decadienal, 4-vinyl-2-methoxyphenol, δ-decalactone, 3-hydroxy-4,5-dimethylfuran-2(5H)-one, dodecanoic acid, and linalool. This study provides insights into the aroma composition of fresh cocoa pulp from different origins for future food applications.

Common Trends and Differences in Antioxidant Activity Analysis of Phenolic Substances Using Single Electron Transfer Based Assays.

Numerous assays were developed to measure the antioxidant activity, but each has limitations and the results obtained by different methods are not always comparable. Popular examples are the DPPH and ABTS assay. Our aim was to study similarities and differences of these two assay regarding the measured antioxidant potentials of 24 phenolic compounds using the same measurement and evaluation methods. This should allow conclusions to be drawn as to whether one of the assays is more suitable for measuring specific subgroups like phenolic acids, flavonols, flavanones, dihydrochalcones or flavanols. The assays showed common trends for the mean values of most of the subgroups. Some dihydrochalcones and flavanones did not react with the DPPH radical in contrast to the ABTS radical, leading to significant differences. Therefore, to determine the antioxidant potential of dihydrochalcone or flavanone-rich extracts, the ABTS assay should be preferred. We found that the results of the flavonoids in the DPPH assay were dependent on the Bors criteria, whereas the structure-activity relationship in the ABTS assay was not clear. For the phenolic acids, the results in the ABTS assay were only high for pyrogallol structures, while the DPPH assay was mainly determined by the number of OH groups.

Mechanisms of Interactions between Bile Acids and Plant Compounds-A Review.

Plant compounds are described to interact with bile acids during small intestinal digestion. This review will summarise mechanisms of interaction between bile acids and plant compounds, challenges in in vivo and in vitro analyses, and possible consequences on health. The main mechanisms of interaction assume that increased viscosity during digestion results in reduced micellar mobility of bile acids, or that bile acids and plant compounds are associated or complexed at the molecular level. Increasing viscosity during digestion due to specific dietary fibres is considered a central reason for bile acid retention. Furthermore, hydrophobic interactions are proposed to contribute to bile acid retention in the small intestine. Although frequently hypothesised, no mechanism of permanent binding of bile acids by dietary fibres or indigestible protein fractions has yet been demonstrated. Otherwise, various polyphenolic structures were recently associated with reduced micellar solubility and modification of steroid and bile acid excretion but underlying molecular mechanisms of interaction are not yet fully understood. Therefore, future research activities need to consider the complex composition and cell-wall structures as influenced by processing when investigating bile acid interactions. Furthermore, influences of bile acid interactions on gut microbiota need to be addressed to clarify their role in bile acid metabolism.

Differentiation of Adsorptive and Viscous Effects of Dietary Fibres on Bile Acid Release by Means of In Vitro Digestion and Dialysis.

To explain the cholesterol-reducing effects of dietary fibres, one of the major mechanisms proposed is the reduced reabsorption of bile acids in the ileum. The interaction of dietary fibres with bile acids is associated with their viscous or adsorptive effects. Since these fibre characteristics are difficult to investigate in vivo, suitable in vitro methodologies can contribute to understanding the mechanistic principles. We compared the commonly used centrifugal approach with a modified dialysis method using dietary fibre-rich materials from different sources (i.e., barley, citrus, lupin, and potato). Digestion was simulated in vitro with oral, gastric, and small intestinal digestion environments. The chyme was dialysed and released bile acids were analysed by high-performance liquid chromatography. The centrifugation method showed adsorptive effects only for cholestyramine (reference material) and a high-fibre barley product (1.4 µmol taurocholic acid/100 mg dry matter). Alternatively, the dialysis approach showed higher values of bile acid adsorption (2.3 µmol taurocholic acid/100 mg dry matter) for the high-fibre barley product. This indicated an underestimated adsorption when using the centrifugation method. The results also confirmed that the dialysis method can be used to understand the influence of viscosity on bile acid release. This may be due to entrapment of bile acids in the viscous chyme matrix. Further studies on fibre structure and mechanisms responsible for viscous effects are required to understand the formation of entangled networks responsible for the entrapment of the bile acids.

Enzymatic esterification of free fatty acids in vegetable oils utilizing different immobilized lipases.

Different immobilized lipases were screened for their ability to esterify free fatty acids (FFA) with monoacylglycerol (MAG) as acyl-group acceptor. A lipase from Rhizomucor miehei (Lipozyme RMIM) was the most suitable for lipase-catalyzed de-acidification-a promising alternative to conventional neutralization. A reduction of the FFA content to 0.6 % (w/w) was achieved by applying a substrate with an initial FFA-content of 6 % (w/w), the reaction at 50 °C for 22 h as well as the stepwise addition of a quadruple stoichiometric amount of MAG.

Antioxidant activity, lipophilicity and extractability of polyphenols from pig skin - development of analytical methods for skin permeation studies.

Permeation of polyphenols through the stratum corneum barrier is a precondition for the protective action of polyphenols against oxidative skin damage. Prior to in vitro skin permeation experiments, we developed a method for the quantification of polyphenols in pig skin, including organic solvent extraction and HPLC analysis. Catechine hydrate, epigallocatechin gallate, trans-resveratrol, quercetin, rutin and protocatechuic acid were chosen for this study as representatives of phenolics with different lipophilicity and molecular weight. The antioxidative activities of polyphenols as well as their octanol-water partition coefficients at different pH values were determined. Extraction of polyphenols from pig skin was optimized by variation of solvent composition, homogenization intensity and time, as well as partial exclusion of oxygen during extraction. The highest recovery rates could be reached by extraction with the methanol-water mixture (90:10, v/v), containing 0.2 g/L l-ascorbic acid, after the cryo-milling for 4 min. Recoveries of 72% for total phenolics, 96% for quercetin and protocatechuic acid, 90% for rutin and 74% for trans-resveratrol, were achieved. These extraction parameters will be selected for the polyphenol extraction from pig skin for further in vitro drug permeation studies.

Physicochemical and chemical properties of mung bean protein isolate affected by the isolation procedure.

The effects of different mung bean protein isolation methods on the chemical composition, the physicochemical properties, and selected antinutritional factors of mung bean protein isolates were investigated. Six protein isolates were prepared by isoelectric precipitation at different extraction pH levels (pH 8 and 9), by micellization, and by hybrid isolation at varying salt concentrations (0.25 M, 0.50 M, 0.75 M). The extraction conditions affected the amount of antinutritive compounds of the isolates. Compared to mung bean flour, micellization reduced phytic acid content by approximately 48% and trypsin inhibitor activity by around 88%. The remaining phytic acid concentration of the isolates influenced their re-solubility, particularly under acidic conditions. The protein isolates exhibited significant differences in surface hydrophobicity and thermal characteristics, indicating structural modifications caused by the extraction methods. Micellization and extraction at pH 8 were identified as mildest isolation methods, as evidenced by the highest enthalpy values. SDS-PAGE analysis demonstrated an enrichment of globulins and comparable protein profiles among the isolates, suggesting that the observed differences arise from conformational changes rather than variations in protein composition. The product yield in protein extraction from mung beans ranged from 8% to 19%, emphasizing the importance of enhancing overall extraction efficiency or exploring the utilization of by-products obtained during the protein isolation process.

Thermal stabilisation of cocoa fruit pulp - Effects on sensory properties, colour and microbiological stability.

To improve cocoa pulp's shelf-life, preservation processes are necessary while maintaining the quality of the pulp. We applied pasteurisation and UHT-treatment and investigated different quality parameters: dry matter content, water activity, total soluble solids, colour and peroxidase activity. Both technologies inactivated peroxidase successfully. The colour of the pasteurised pulp was similar to the fresh, while UHT-treated pulp was more brownish. The sensory properties were investigated in detail by descriptive analysis and the identification of aroma-active volatile organic compounds. Fresh pulp revealed the highest aroma intensity for attribute unripe banana-like, whereas UHT-treated pulp scored highest in the intensity of attribute tropical fruit-like. Pasteurised pulp showed strong similarities to the fresh pulp. Fresh cocoa pulp exhibited 74 aroma-active regions identified by GC-MS/O. UHT-treated and pasteurised pulp accounted for 66 and 60 aroma-active regions, respectively. Five identified substances were only found in the fresh and pasteurised pulp, namely: δ-carene, 1-pentanol, 3-(methylthio)propanol, phenol and δ-undecalactone. Similarly, fresh and UHT-treated pulp shared ten exclusive odorants, such as decanal, geraniol, and δ-nonalactone. The pasteurised and UHT-treated pulp shared two compounds, δ-decalactone and 5-(hydroxymethyl)furfural. Furthermore, the thermally treated pulps could be stored at 4 °C and 23 °C for 24 weeks without observing a significant growth of microorganisms. The rate of non-enzymatic browning was higher in samples stored at 23 °C compared to those stored at 4 °C, leading to higher browning indices. We demonstrated that pasteurisation and ultra-high temperature treatment are suitable technologies for the stabilisation of cocoa fruit pulp. These resulted in prolonged shelf-lifes and minimal changes in the sensory prorperties of the treated pulps, characterised by a reduction in the aroma diversities. This work provides important insights for the thermal stabilisation of further side-streams.

Radical Scavenging Mechanisms of Phenolic Compounds: A Quantitative Structure-Property Relationship (QSPR) Study.

Due to their antioxidant properties, secondary plant metabolites can scavenge free radicals such as reactive oxygen species and protect foods from oxidation processes. Our aim was to study structural influences, like basic structure, number of hydroxyl groups and number of Bors criteria on the outcome of the oxygen radical absorbance capacity (ORAC) assay. Furthermore, similarities and differences to other in vitro antioxidant assays were analyzed by principal component analysis. Our studies confirmed that the antioxidant behavior in the ORAC assay is dominated by the number and types of substituents and not by the Bors criteria, as long as no steric hindrance occurs. For example, morin (MOR) with five hydroxyl groups and two Bors criteria reached an area under the curve of (3.64 ± 0.08) × 105, which was significantly higher than quercetin-7-D-glucoside (QGU7) (P < 0.001), and thus the highest result. Principal component analysis showed different dependencies regarding structural properties of Folin-Ciocalteu (FC)- and 2,2-diphenyl-1-picrylhydrazyl (DPPH)-assays or 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS)- and ORAC-assays, respectively. Therefore, we conclude that they are based on different reaction mechanisms. The number of hydroxyl groups showed a stronger influence on the antioxidant activity than the Bors criteria. Due to these differences, the correlation of these rapid tests to specific applications should be validated.

Aroma-active volatiles and rheological characteristics of the plastic mass during conching of dark chocolate.

Chocolate conching is a highly complex, thermomechanical process that transforms the aroma and flow properties of a dry starting material. Different conched plastic masses of dark chocolate were characterized. Rheological characterization of plastic masses was performed for the first time using a closed cavity rheometer (CCR1). In free cocoa butter derived from the plastic masses, acetic acid, benzaldehyde, (R,S)-(±)-linalool, 2,3,5,6-tetramethylpyrazine, and 2-phenylethanol were quantified by stable isotope dilution analysis (SIDA2) and gas chromatography-mass spectrometry. During the conching process, the amount of free cocoa butter increased possibly due to de-agglomeration. The complex viscosity of the plastic mass decreased as a function of conching time. Regarding aroma refinement, the concentrations of all five aroma-active volatiles decreased with increasing conching duration, albeit to varying degrees. The level of acetic acid showed the most pronounced decrease of about 60%, whereas linalool exhibited the lowest decrease in concentration, up to 26%. Overall, a lower polarity or boiling point of the aroma-active volatiles was linked to a stronger decrease in concentration during conching. These data illustrate the influence of conching on texture and the respective aroma changes, which deepens understanding of the conching effect on the sensory quality of dark chocolate.

Nutritional Composition of Beach-Cast Marine Algae from the Brazilian Coast: Added Value for Algal Biomass Considered as Waste.

In some coastal areas, large quantities of beach-cast macroalgae can accumulate and are usually considered waste and disposed of. However, due to their biofunctional and nutritional properties, they have great potential as a new source of raw materials. Increasing population growth has made the search for alternative raw materials with valuable nutritional properties urgent; here, beach-cast macroalgae could provide great potential. Our research goal was to characterize the nutritional profile of 12 beach-cast seaweed species from the Brazilian coast to assess their potential valorization. A considerable number of nutritional compounds was observed, such as ash (6.5-59.3%), total dietary fibers (22.1-65.8%), proteins (5.1-21.5%), and carbohydrates (31.4-81.0%), with an expressive abundance of minerals, free amino acids, and fatty acids. Spatoglossum schroederi and Alsidium seaforthii showed protein contents of 21.5 ± 0.2%, 19.7 ± 0.1%, and high amounts of total dietary fiber of 59.2 ± 0.4%, 61.7 ± 4.9%, respectively. The overall profile suggests that beach-cast seaweeds are suitable for nutritional and other bioeconomical purposes, to which different species with different characteristics contribute. Contamination of these seaweeds with unwanted toxic compounds like micropollutants was not studied. However, this must be considered before they are used for human consumption.

Enzymatic Hydrolysis and Fermentation of Pea Protein Isolate and Its Effects on Antigenic Proteins, Functional Properties, and Sensory Profile.

Combinations of enzymatic hydrolysis using different proteolytic enzymes (papain, Esperase®, trypsin) and lactic fermentation with Lactobacillus plantarum were used to alter potential pea allergens, the functional properties and sensory profile of pea protein isolate (PPI). The order in which the treatments were performed had a major impact on the changes in the properties of the pea protein isolate; the highest changes were seen with the combination of fermentation followed by enzymatic hydrolysis. SDS-PAGE, gel filtration, and ELISA results showed changes in the protein molecular weight and a reduced immunogenicity of treated samples. Treated samples showed significantly increased protein solubility at pH 4.5 (31.19-66.55%) and at pH 7.0 (47.37-74.95%), compared to the untreated PPI (6.98% and 40.26%, respectively). The foaming capacity was significantly increased (1190-2575%) compared to the untreated PPI (840%). The treated PPI showed reduced pea characteristic off-flavors, where only the treatment with Esperase® significantly increased the bitterness. The results from this study suggest that the combination of enzymatic hydrolysis and lactic fermentation is a promising method to be used in the food industry to produce pea protein ingredients with higher functionality and a highly neutral taste. A reduced detection signal of polyclonal rabbit anti-pea-antibodies against the processed protein preparations in ELISA furthermore might indicate a decreased immunological reaction after consumption.

Recovery, Isolation, and Characterization of Food Proteins.

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Contribution of S. xylosus and L. sakei ssp. carnosus Fermentation to the Aroma of Lupin Protein Isolates.

Aroma-active compounds of lupin protein isolate and lupin protein isolate fermented with Staphylococcus xylosus and Lactobacillus sakei ssp. carnosus were investigated. The changes in aroma-active compounds were determined by application of aroma extract dilution analysis in combination with gas chromatography-mass spectrometry/olfactometry for identification, and by stable isotope dilution assays for quantification. A total of 30 aroma-active compounds for non-fermented and fermented samples were identified. The aroma profile of LPI fermented with Lactobacillus sakei ssp. carnosus was characterized as roasty and popcorn-like. Staphylococcus xylosus generated cheesy impressions, being in line with the fact that the main aroma compounds acetic acid, butanoic acid, and 2/3-methylbutanoic acid could be identified. Quantification of butanoic acid further confirmed these findings with the highest concentration of 140 mg/kg for LPI fermented with Staphylococcus xylosus. Our study provides insights into how fermentation utilizing different fermentative microbial strains, namely Staphylococcus xylosus and Lactobacillus sakei ssp. carnosus alters the aroma profile of lupin protein isolates. This demonstrates the potential of shaping fermented protein-based foods via targeted microbiological refinement.

How Does the Phenol Structure Influence the Results of the Folin-Ciocalteu Assay?

Plants produce a diverse array of secondary metabolites that are generally nonessential but facilitate ecological interactions. Fruits, vegetables, seeds and nuts can accumulate bioactive secondary metabolites with health-promoting properties, including the potent antioxidant activities of phenolic compounds. Several in vitro assays have been developed to measure the polyphenol content and antioxidant activity of plant extracts, e.g., the simple and highly popular Folin-Ciocalteu (FC) assay. However, the literature contains a number of different descriptions of the assay and it is unclear whether the assay measures the polyphenol content or reducing capacity of the sample. To determine the influence of phenolic structures on the outcome of the FC assay, we tested phenols representing different subgroups (phenolic acids, flavonols, flavanols, dihydrochalcones and flavanones). We observed different results for each reference substance and subgroup. Accordingly, we concluded that the FC assay does not measure the polyphenol content of a sample but determines its reducing capacity instead. Assigning the substances to five structural classes showed that the FC results depend on the number of fulfilled Bors criteria. If a molecule fulfills none of the Bors criteria, the FC results depend on the number of OH groups. We did not find a correlation with other single electron transfer assays (e.g., ABTS and DPPH assays). Furthermore, the FC assay was compatible with all five subgroups and should be preferred over the DPPH assay, which is specific for extracts rich in dihydrochalcones or flavanones.

Fermentation of Lupin Protein Hydrolysates-Effects on Their Functional Properties, Sensory Profile and the Allergenic Potential of the Major Lupin Allergen Lup an 1.

Lupin protein isolate was treated using the combination of enzymatic hydrolysis (Papain, Alcalase 2.4 L and Pepsin) and lactic acid fermentation (Lactobacillus sakei ssp. carnosus, Lactobacillus amylolyticus and Lactobacillus helveticus) to investigate the effect on functional properties, sensory profile and protein integrity. The results showed increased foaming activity (2466-3481%) and solubility at pH 4.0 (19.7-36.7%) of all fermented hydrolysates compared to the untreated lupin protein isolate with 1613% of foaming activity and a solubility of 7.3 (pH 4.0). Results of the SDS-PAGE and Bead-Assay showed that the combination of enzymatic hydrolysis and fermentation of LPI was effective in reducing L. angustifolius major allergen Lup an 1 to a residual level of <0.5%. The combination of enzymatic hydrolysis and fermentation enables the production of food ingredients with good functional properties in terms of protein solubility and foam formation, with a balanced aroma and taste profile.

Sensory profile, functional properties and molecular weight distribution of fermented pea protein isolate.

Pea protein isolate (PPI, from Pisum sativum L.) was fermented with six different lactic acid bacteria strains for 24 â€‹h and 48 â€‹h. The fermented samples were analyzed regarding their retronasal aroma and taste, their protein solubility, emulsifying and foaming capacity. Changes in the molecular weight distribution were analyzed to monitor potential effects of fermentation on the main allergenic protein fractions of PPI. After 24-h fermentation, PPI's characteristic aroma attributes and bitter taste decreased for all fermented PPI. However, after 48-h fermentation, cheesy aroma, and acid and salty tastes were increased. The PPI fermented with L. plantarum showed the most neutral taste and the panel's highest preference; instead, fermentation with L. fermentum led to a fecal aroma and was the least preferred. The protein solubility and emulsifying capacity decreased after PPI fermentation, while foaming capacity remained constant in comparison to the untreated PPI. The electrophoretic results showed a reduction in the intensity of the allergenic protein fractions; however, these changes might be attributed to the reduced protein solubility rather than to a high proteolytic effect of the strains. Fermentation of PPI for 24 â€‹h and 48 â€‹h might not be a suitable method for the production of highly functional pea proteins. Further modification methods have to be investigated in the future.

Screening of Twelve Pea (Pisum sativum L.) Cultivars and Their Isolates Focusing on the Protein Characterization, Functionality, and Sensory Profiles.

Pea protein concentrates and isolates are important raw materials for the production of plant-based food products. To select suitable peas (Pisum sativum L.) for protein extraction for further use as food ingredients, twelve different cultivars were subjected to isoelectric precipitation and spray drying. Both the dehulled pea flours and protein isolates were characterized regarding their chemical composition and the isolates were analyzed for their functional properties, sensory profiles, and molecular weight distributions. Orchestra, Florida, Dolores, and RLPY cultivars showed the highest protein yields. The electrophoretic profiles were similar, indicating the presence of all main pea allergens in all isolates. The colors of the isolates were significantly different regarding lightness (L*) and red-green (a*) components. The largest particle size was shown by the isolate from Florida cultivar, whereas the lowest was from the RLPY isolate. At pH 7, protein solubility ranged from 40% to 62% and the emulsifying capacity ranged from 600 to 835 mL g-1. The principal component analysis revealed similarities among certain pea cultivars regarding their physicochemical and functional properties. The sensory profile of the individual isolates was rather similar, with an exception of the pea-like and bitter attributes, which were significantly different among the isolates.

Effect of Physicochemical Properties of Carboxymethyl Cellulose on Diffusion of Glucose.

Soluble dietary fibers (SDF) are known to reduce the post-prandial plasma glucose levels. However, the detailed mechanisms of this reduced glucose release in the human gut still remain unclear. The aim of our study was to systematically investigate the effect of different types of SDF on glucose release in an in vitro model as a prerequisite for the selection of fibers suitable for application in humans. Three types of carboxymethyl cellulose (CMC) were used to investigate the correlations between fiber concentration, molecular weight (MW), and viscosity on diffusion of glucose using a side-by-side system. CMC solutions below the coil overlap (c*) influenced the glucose diffusivity only marginally, whereas at concentrations above c* the diffusion of glucose was significantly decreased. Solutions of lower MW exhibited a lower viscosity with lower glucose diffusion compared to solutions with higher MW CMC, attributed to the higher density of the solutions. All CMC solutions showed a systematic positive deviation from Stokes-Einstein behavior indicating a greater rise in viscosity than reduction in diffusion. Therefore, our results pave the way for a new approach for assessing glucose diffusion in solutions comprising dietary fibers and may contribute to further elucidating the mechanisms of post-prandial plasma glucose level reduction.

Characterisation of the molecular interactions between primary bile acids and fractionated lupin cotyledons (Lupinus angustifolius L.).

Interactions between bile acids and plant-based materials, and the related feedback mechanisms in enterohepatic circulation, have been considered targets for lowering cholesterol. This study aimed to identify lupin compounds that interact with primary bile acids on molecular level. Lupin cotyledons were fractionated and bile acid adsorbing activities were investigated using in vitro digestion, equilibrium dialysis and kinetic analyses. Protein- and fibre-enriched fractions significantly (p ≤ 0.05) adsorbed chenodesoxycholic acids (up to 2.33 µmol/100 g DM). Alcohol purification showed that bile acid adsorption is independent of protein and fibre structures. Moreover, high adsorption was observed with an alcohol extract (6.97 µmol chenodesoxycholic acids/100 g DM) that was rich in phytochemicals, such as flavonoids (1842 mg/100 g DM). These results suggest the formation of hydrophobic interactions between polyphenols and bile acids. Further studies of molecular mechanisms are required to define the contributions of polyphenols to the cholesterol-lowering actions of lupins.