An in-depth multiphasic analysis of the chocolate production chain, from bean to bar, demonstrates the superiority of Saccharomyces cerevisiae over Hanseniaspora opuntiae as functional starter culture during cocoa fermentation.

Hanseniaspora opuntiae is a commonly found yeast species in naturally fermenting cocoa pulp-bean mass, which needed in-depth investigation. The present study aimed at examining effects of the cocoa isolate H. opuntiae IMDO 040108 as part of three different starter culture mixtures compared with spontaneous fermentation, regarding microbial community, substrate consumption, and metabolite production dynamics, including volatile organic compound (VOC) and phytochemical compositions, as well as compositions of the cocoa beans after fermentation, cocoa liquors, and chocolates. The inoculated H. opuntiae strain was unable to prevail over background yeasts present in the fermenting cocoa pulp-bean mass. It led to under-fermented cocoa beans after four days of fermentation, which was however reflected in higher levels of polyphenols. Cocoa fermentation processes inoculated with a Saccharomyces cerevisiae strain enhanced flavour production during the fermentation and drying steps, which was reflected in richer and more reproducible aroma profiles of the cocoa liquors and chocolates. Sensory analysis of the cocoa liquors and chocolates further demonstrated that S. cerevisiae led to more acidic notes compared to spontaneous fermentation, as a result of an advanced fermentation degree. Finally, different VOC profiles were found in the cocoa beans throughout the whole chocolate production chain, depending on the fermentation process.

Inhibitory Effect on Nitric Oxide Release in LPS-Stimulated Macrophages and Free Radical Scavenging Activity of Croton linearis Jacq. Leaves.

Oxidative stress is an important component of many diseases including cancer, along with inflammatory and neurodegenerative processes. Natural antioxidants have emerged as promising substances to protect the human body against reactive oxygen and nitrogen species. The present study evaluates the inhibition of nitric oxide (NO) production in LPS-stimulated RAW 264.7 murine macrophages and the free radical scavenging activity of Croton linearis Jacq. leaves. UPLC-QTOF-MS analysis identified 18 compounds: nine alkaloids with a morphinane, benzylisoquinoline or aporphine nucleus, and nine O-glycosylated-flavonoids with quercetin, kaempferol and isorhamnetin as the aglycones. The crude extract (IC50 21.59 µg/mL) and the n-hexane fraction (IC50 4.88 µg/mL) significantly reduced the NO production in LPS-stimulated macrophages but with relatively high cytotoxicity (CC50 75.30 and CC50 70.12 µg/mL, respectively), while the ethyl acetate fraction also showed good activity (IC50 40.03 µg/mL) without affecting the RAW 264.7 cell viability. On the other hand, the crude extract, as well as the dichloromethane and ethyl acetate fractions, showed better DPPH and ABTS free radical scavenging activities. Considering the chemical composition and the activity observed for Croton linearis leaves, they may be considered a good source of antioxidants to combat oxidative damage-related diseases.

Acute Effects of Cocoa Flavanols on Blood Pressure and Peripheral Vascular Reactivity in Type 2 Diabetes Mellitus and Essential Hypertension.

BACKGROUND: Type 2 diabetes mellitus (T2DM) is associated with a high risk of vascular complications. Interestingly, cocoa flavanols (CF) can exert beneficial vascular effects in non-diabetic subjects. However, these effects have only been scarcely studied in T2DM. Therefore, we performed a study to assess the effects on vascular reactivity of a single dose of CF (790 mg) in T2DM and whether certain antihypertensive drugs may modulate these effects. METHODS: 24 non-diabetic and 11 T2DM subjects were studied in a cross-over design. Fasting blood samples, blood pressure (BP), and arterial vasoreactivity (flow-mediated dilation) were assessed before and 70 min after capsule ingestion. Muscle microvascular reactivity was only assessed after capsule ingestion. Age, waist-to-hip ratio, BP at baseline, and the use of antihypertensive drugs were regarded as covariates in a mixed models analysis. RESULTS: CF ingestion did not affect any parameter. However, independent of the type of capsules ingested, a decrease in diastolic BP by 3 mmHg (95% CI: -4.0; -2.0) and an increase in the change in brachial artery diameter (pre vs. post occlusion) by 0.06 mm (95% CI: 0.01; 0.12) were detected in the non-diabetic group, while they remained unchanged in the T2DM group. CONCLUSION: No beneficial effects of CF were detected on vascular reactivity parameters in T2DM and non-diabetic participants.

The effect of cocoa alkalization on the non-volatile and volatile mood-enhancing compounds.

Alkalization is a process to improve color, dispersibility and flavor of cocoa powder but is likely to have a negative effect on the phytochemicals. Hereto, the impact of alkalization degree (none, medium and high) on the potential mood-enhancing compounds corresponding to the four levels of the mood pyramid model (flavanols, methylxanthines, biogenic amines and orosensory properties) was investigated. The phytochemical content, analyzed via UPLC-HRMS, showed reduction of specific potential mood-enhancing compounds upon alkalization, implying a decrease in bitterness and astringency. Moreover, volatile compounds analysis via HS-SPME-GC-MS indicated that alkalization reduced the levels of volatile compounds, responsible for acidity, fruity, floral and cocoa aromas. With respect to the orosensory properties, the cocoa powder palatability was suggested to be increased due to reduced acidity, bitterness, and astringency, while the desired volatile compounds were reduced. However, sensorial analysis is required to link the volatile results with the overall effect on the flavor perception.

Phytochemical composition and antioxidant activity of Cinnamomum burmannii Blume extracts and their potential application in white chocolate.

This study aims at determining the potentials of cinnamon (Cinnamomun burmannii) extracts to improve the health-promoting properties of white chocolate. LC-HRMS analysis was employed to obtain information regarding the phytochemical content while the phosphomolybdenum, FRAP and DPPH assays were used to determine antioxidant activity of cinnamon extract. Furthermore, the cinnamon extract was loaded into nanoparticles before adding it to white chocolate. The results show that cinnamon extracts contained phenols up to 310 mg EE and possessed antioxidant activity up to 260 mg TAE per gram of dry extract depending on the extraction mode (i.e., traditional and ultrasonic-assisted method) and the solvent type. The cinnamon extract contained catechin, epicatechin, procyanidin B2, quercitrin, 3,4-dihydroxybenzaldehyde, protocatechuic acid and cinnamic acid at levels of 51, 53, 1396, 13, 1138, 228 and 934 µg/g of dry extract, respectively. The encapsulated cinnamon extract increased the phenolic content of white chocolate from 47.6 to 1060.6 µg EE/g.

Roasting-induced changes in cocoa beans with respect to the mood pyramid.

The mood pyramid of cocoa, which was previously proposed as a new concept, consists of four levels (flavan-3-ols, methylxanthines, minor compounds and orosensory properties). Roasting is a crucial process for flavor development in cocoa but is likely to have a negative impact on the phytochemicals. We investigated the effect of roasting time (10-50 min) and temperature (110-160 °C) on the potential mood-enhancing compounds corresponding to the distinct mood pyramid levels. Phytochemicals were analyzed using UPLC-HRMS, while the flavor was mapped via aroma (HS-SPME-GC-MS) and generic descriptive analysis (trained panel). Results revealed that roasting at 130 °C for 30 min did not significantly affect the levels of epicatechin, procyanidin B2 and theobromine, while salsolinol significantly increased. Moreover, bitterness and astringency were reduced and the desired cocoa flavor was developed. Thus, through selection of appropriate roasting time and temperature conditions phytochemicals of interest could be retained without comprising the flavor.

In Vitro Anti-Inflammatory, Anti-Oxidant, and Cytotoxic Activities of Four Curcuma Species and the Isolation of Compounds from Curcuma aromatica Rhizome.

The genus Curcuma is part of the Zingiberaceae family, and many Curcuma species have been used as traditional medicine and cosmetics in Thailand. To find new cosmeceutical ingredients, the in vitro anti-inflammatory, anti-oxidant, and cytotoxic activities of four Curcuma species as well as the isolation of compounds from the most active crude extract (C. aromatica) were investigated. The crude extract of C. aromatica showed 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity with an IC50 value of 102.3 µg/mL. The cytotoxicity effect of C. aeruginosa, C. comosa, C. aromatica, and C. longa extracts assessed with the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay at 200 µg/mL were 12.1 2.9, 14.4 4.1, 28.6 4.1, and 46.9 8.6, respectively. C. aeruginosa and C. comosa presented apoptosis cells (57.7 3.1% and 32.6 2.2%, respectively) using the CytoTox-ONE™ assay. Different crude extracts or phytochemicals purified from C. aromatica were evaluated for their anti-inflammatory properties. The crude extract of C. aromatica showed the highest potential to inhibit NF-κB activity, followed by C. aeruginosa, C. comosa, and C. longa, respectively. Among the various purified phytochemicals curcumin, germacrone, curdione, zederone, and curcumenol significantly inhibited NF-κB activation in tumor necrosis factor (TNF) stimulated HaCaT keratinocytes. Of all compounds, curcumin was the most potent anti-inflammatory.

Development of a classification model for the antigenotoxic activity of flavonoids.

Genotoxic agents are capable of causing damage to genetic material and the cumulative DNA damage causes mutations, involved in the development of various pathological conditions, including cancer. Antigenotoxic agents possess the potential to counteract these detrimental cellular modifications and may aid in preventing, delaying, or decreasing the severity of these pathological conditions. An important class of natural products for which promising antigenotoxic activities have already been shown, are the flavonoids. In this research, we investigated the quantitative structure-activity relationship (QSAR) of flavonoids and their antigenotoxic activity against benzo[a]pyrene (B[a]P) and its mutagenic metabolite B[a]P-7,8-diol-9,10-epoxide-2. Random Forest classification models were developed, which could be useful as a preliminary in silico evaluation tool, before performing in vitro or in vivo experiments. The descriptors G2S and R8s. were the most significant for predicting the antigenotoxic potential.

Non-volatile and volatile composition of West African bulk and Ecuadorian fine-flavor cocoa liquor and chocolate.

Cocoa products are obtained from the seeds of Theobroma cacao L. In this research, cocoa liquor and chocolate produced from cocoa beans from West Africa (Forastero, "bulk" cacao) and Ecuador (Nacional variety, "fine-flavor" cacao), were investigated, using a novel approach in which various analytical techniques are combined in order to obtain in-depth knowledge of the studied cocoa samples. The levels of various classes of primary metabolites were determined and a wide range of secondary metabolites, including volatile organic acids, aldehydes, esters, pyrazines, polyphenols, methylxanthines and biogenic amines, were identified and/or quantified by HS-SPME GC-MS (headspace-solid phase microextraction gas chromatography - mass spectrometry). and UPLC-HRMS (ultra-performance liquid chromatography - high resolution mass spectrometry). Odor Activity Values (OAV) were calculated to assess the contribution of individual volatiles on the final aroma. Various volatile aroma compounds were more abundant in the West African cocoa liquor and chocolate, while the Ecuadorian samples were richer in most quantified non-volatile metabolites. Principal component analysis (PCA) confirmed that the four samples can be clearly distinguished. Alcohols, pyrazines, amino acids and biogenic amines were found to be highly influential in causing this differentiation. The proposed approach can be useful in future studies on more extensive cocoa sample collections, in order to highlight similarities and pinpoint typical differences in chemical composition among these samples.

Antimicrobial activity of leaf extracts and isolated constituents of Croton linearis.

ETHNOPHARMACOLOGICAL RELEVANCE: Leaves of Croton linearis, known as "rosemary", are widely used in folk medicine in Caribbean countries to treat fever and colds (associated to infections). AIM OF THE STUDY: To evaluate the in vitro antimicrobial activity of extracts and fractions derived from C. linearis leaves. MATERIALS AND METHODS: Bioassay-guided fractionation and isolation of compounds from an ethanolic extract of C. linearis, using flash chromatography and semi-preparative HPLC-DAD-MS (High Performance Liquid Chromatography - Diode Array Detection - Mass Spectrometry). Isolated compounds were characterized by MS, 1D- and 2D-NMR (Nuclear Magnetic Resonance) spectroscopy. The microdilution method with resazurin, as well as direct counting with an optical microscope, were used to determine the in vitro antimicrobial activity against bacteria, fungi and parasites. Moreover, the cytotoxicity on human fetal lung fibroblasts (MRC-5) was evaluated. RESULTS: The total extract and chloroform fraction (ClF) showed high activity against protozoa with IC50 values ranging from 1 to 26 µg/mL, but also cytotoxicity on MRC-5 and PMM (Peritoneal Murine Macrophages). Seven compounds were isolated and characterized for first time in this species: the alkaloids laudanidine, laudanosine, reticuline, corydine, glaucine and cularine and the flavonoid glycoside isorhamnetin-3-O-(6″O-p-trans-coumaroyl)-ß-glucopyranoside. Reticuline showed a weak activity against L. infantum (IC50 148.0 ±â€¯1.2 µM), while the flavonoid was active against T. cruzi (IC50 35.6 ±â€¯2.3 µM). CONCLUSIONS: The results show the antiprotozoal potential of the extract and some isolated constituents, which supports the use of this species in Caribbean folk medicine.

Phytochemical characterization and comparative studies of four Cecropia species collected in Panama using multivariate data analysis.

Plant species of the genus Cecropia (Urticaceae) are used as traditional medicine in Latin-America, and are commercially available as food supplements. The aim of this study was to characterize and compare the phytochemical constituents of four Cecropia species collected in Panama. The structures of 11 compounds isolated from leaves of C. obtusifolia were elucidated based on high resolution mass spectrometry (HRMS) and nuclear magnetic resonance (NMR) spectroscopic analysis; the polyphenolic constituents of leaves of all four Cecropia species and commercial products were characterized using high performance liquid chromatography-diode array detection-quadrupole time of flight-tandem high resolution mass spectrometry (HPLC-DAD-QTOF). Forty-seven compounds were fully identified or tentatively characterized. Thirty-nine of these have not been previously reported for the species under investigation. Multivariate analysis revelead that C. obtusifolia and C. insignis are the most related species, while C. hispidissima is the most segregated one. Considering the importance of the description of novel chemical entities and the increasing interest and use of natural products, this study may be of great help for chemotaxonomic purposes, the interpretation of medicinal properties and for quality assessment of herbal supplements containing Cecropia leaves.