Changes in GC-MS metabolite profile, antioxidant capacity and anthocyanins content during fermentation of fine-flavor cacao beans from Ecuador.

The fermentation of fine-flavor cacao beans is a key process contributing to the enhancement of organoleptic attributes and monetary benefits for cacao farmers. This work aimed to describe the dynamics of the gas chromatography-mass spectrometry (GC-MS) metabolite profile as well as the antioxidant capacity and anthocyanin contents during fermentation of fine-flavor cacao beans. Samples of Nacional x Trinitario cacao beans were obtained after 0, 24, 48, 72, 96, and 120 hours of spontaneous fermentation. Total phenolic content (TPC), ferric reducing antioxidant power (FRAP), and total anthocyanin content were measured by ultraviolet-visible (UV-Vis) spectrophotometry. Volatiles were adsorbed by headspace solid phase microextraction (HS-SPME) while other metabolites were assessed by an extraction-derivatization method followed by gas chromatography-mass spectrometry (GC-MS) detection and identification. Thirty-two aroma-active compounds were identified in the samples, including 17 fruity, and 9 floral-like volatiles as well as metabolites with caramel, chocolate, ethereal, nutty, sweet, and woody notes. Principal components analysis and Heatmap-cluster analysis of volatile metabolites grouped samples according to the fermentation time. Additionally, the total anthocyanin content declined during fermentation, and FRAP-TPC values showed a partial correlation. These results highlight the importance of fermentation for the improvement of the fine-flavor characteristics of cacao beans.

Chemical Characterization of Trichoderma spp. Extracts with Antifungal Activity against Cocoa Pathogens.

Ecuador is one of the major cocoa producers worldwide, but its productivity has lately been affected by diseases. Endophytic biocontrol agents have been used to minimize pathogenic effects; however, compounds produced by endophytes are minimally understood. This work presents the chemical characterization of the Trichoderma species extracts that proved inhibition against cocoa pathogens. Solid-liquid extraction was performed as a partitioning method using medium with the fungal mycelia of Trichoderma reesei (C2A), Trichoderma sp. (C3A), Trichoderma harzianum (C4A), and Trichoderma spirale (C10) in ethyl acetate individually. The extract of T. spirale (C10) exhibited the growth inhibition (32.97-47.02%) of Moniliophthora perniciosa at 10 µg/mL, while a slight stimulation of Moniliophthora roreri was shown by the extracts of T. reesei (C2A) and T. harzianum (C4A) at higher concentrations. The inhibitory activity could be related to alkaloids, lactones, quinones, flavonoids, triterpenes, and sterols, as indicated by chemical screening and antifungal compounds, such as widdrol, ß-caryophyllene, tyrosol, butyl isobutyrate, sorbic acid, palmitic acid, palmitelaidic acid, linoleic acid, and oleic acid, which were identified by gas chromatography-mass spectrometry (GC-MS). The results showed that the extracts, particularly T. spirale (C10), have the potential as biocontrol agents against witches' broom disease; however, further studies are needed to confirm their effectiveness.

Anti-inflammatory potential of processing Vernonanthura patens (Kunth) H. Rob. leaves aqueous extract.

Folk medicine uses decoctions of Vernonanthura patens (Kunth) H. Rob. leaves for healing wounds, and moderate pains. In this study, anti-inflammatory activity of decocted aqueous extract and its fractions is discussed. The fractions were obtained by liquid-liquid extraction in a separating funnel with solvents of increasing polarity: hexane, chloroform, and ethyl acetate. Antioxidant capacity, COX1, and COX2 cyclooxygenase inhibitory activities of aqueous extract (A1), aqueous (A2), and ethyl acetate (A3) fractions were assessed. A3 revealed the highest flavonoid content, and DPPH (2,2-diphenyl-1-picrylhydrazyl) scavenging activity. Nevertheless, no significance differences were observed between IC50 values of A1 and A2, and A1 showed anti-inflammatory activity with potential selectivity against COX2 enzyme, but intermediate COX1 inhibition. Further experiments are required to complement the remarkable anti-inflammatory effect of assessed aqueous extract. These results support the medicinal use of this plant species and indicate that A1 can be used as raw material for prospective nutraceutical products.

Effect of solvent-solvent partition on antioxidant activity and GC-MS profile of Ilex guayusa Loes. leaves extract and fractions.

The use of guayusa (Ilex guayusa Loes.) leaves as functional food has increase recently. This work discusses the antioxidant activity and volatile compounds of guayusa leaves extract and fractions. The methanol crude extract was obtained by maceration, subsequently hexane, chloroform, ethyl acetate, and aqueous fractions were collected by solvent-solvent partition. Total phenolic content (TPC), total flavonol/flavone content (TFC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, and ferric reducing antioxidant power (FRAP) were measured by ultraviolet-visible (UV-Vis) spectrophotometry. The results revealed that ethyl acetate fraction showed highest inhibition against DPPH radical (93.86 ± 0.95%) at 500 µg/mL, and reduce the ferric-tripyridyltriazine complex (Fe3+-TPTZ) at 1619.81 mg trolox equivalent (TE)/g, followed by aqueous fraction. This bioactivity could be related to phenolic acids, flavones and flavonols content, as well as the caffeine, dodecanoic acid isopropyl ester, caffeic acid, and malic acid identified by gas chromatography-mass spectrometry (GC-MS). These findings support the antioxidant properties of this plant material.

ITS1 Barcode and Phytochemical Analysis by Gas Chromatography-Mass Spectrometry of Corynaea crassa Hook. f (Balanophoraceae) from Ecuador and Peru.

The use of medicinal plants is the basis of traditional healthcare. Recently, the use of herbal medicine has been increasing among consumers due to availability, economy, and less side effect. For instance, the hemiparasite plant Corynaea crassa has medicinal properties and could be found in some regions of America, from Costa Rica to Bolivia. Phytochemical and genetic characterization of medicinal plants is needed for proper identification of metabolites responsible for medicinal properties and for genotyping, respectively. Moreover, characterization of medicinal plants through the use of DNA barcodes is an important tool for phylogenetic analysis and identification of species; furthermore, complemented with phytochemical analysis, both are useful for identification of plant species and quality control of medicinal products. The objective of this study was to analyze the species of C. crassa collected in Ecuador and Peru from the phylogenetic and phytochemical point of view. Polymerase chain reaction (PCR) was performed for amplification of the internal transcribed spacer 1 (ITS1) region after DNA extraction of samples of C. crassa. Blast analysis was performed in the GenBank database with the ITS1 sequences obtained from two accessions of C. crassa from Ecuador (GenBank accession numbers OM471920 and OM471919 for isolates CIBE-17 and CIBE-18, respectively) and three from Peru (GenBank accession numbers OM471921, OM471922, and OM471923 for isolates CIBE-13, CIBE-14, and CIBE-15, respectively). The accessions available in the GenBank were used for phylogenetic analysis. For the phytochemical analysis, hydroalcoholic extracts were obtained by maceration using 80% ethanol as solvent, followed by a derivatization process and analysis by gas chromatography-mass spectrometry. Based on the phylogenetic analysis of the C. crassa samples, the ITS1 sequence could be used to differentiate C. crassa of different locations. The samples of C. crassa from Ecuador and Peru are more similar between them than with other clades including Helosis spp. The phytochemical study revealed differences in the presence and relative abundance of some metabolites; mainly eugenol, 1,4-lactone arabinonic acid, dimethoxyrabelomycin and azelaic acid, which are reported for the first time for the species under study and the genus Corynaea. These results are the first findings on the combined analysis using genetic and phytochemical analysis for C. crassa, which could be used as a useful tool for quality control of the C. crassa species in medicinal products.

Effect of Cocoa Bean Shell Addition on Metabolite Profile and Antioxidant Activity of Herbal Infusions.

Cocoa bean shell (CBS) is a by-product with aromatic characteristics that can enhance the aroma and bioactivity of herbal infusions. This study was aimed to determine the effect of the addition of cocoa bean shell on the metabolite profile and antioxidant activity of infusions made with Ilex guayusa and Vernonanthura patens and their mixtures. Metabolite profile was analyzed by gas chromatography-mass spectrometry combined with multivariate analysis. Total polyphenol content and flavonoids were determined by the Folin-Ciocalteu method and by the flavonoid-AlCl3 complex, respectively. Antioxidant activities were measured by the decolorization assay of the 2,2-diphenyl-1-picrylhydrazyl radical and the ferric reducing antioxidant power. The results revealed that the addition of CBS increases the content of phenolic acids in the infusions (caffeic acid, 4-hydroxybenzoic acid, and pyrocatechol). Nonetheless, the antioxidant activity of the infusions decreased with the addition of CBS (16.21 to 2.74 TEAC). Carboxylic acids and derivatives, major compounds present in the infusions prepared with V. patens, were the metabolites that showed the highest correlation with the antioxidant activity. This study suggests that the infusions made with CBS present a profile of metabolites different from the infusions of I. guayusa, V. patens, and their mixtures.

Identification of lupeol produced by Vernonanthura patens (Kunth) H. Rob. leaf callus culture.

The lupeol detection in callus of Vernonanthura patens (Kunth) H. Rob. leaves is discussed. Leaf segments previously treated with sodium hypochlorite, ethanol, and distilled water were placed in MS basal medium (Murashige and Skoog) for 7 days. Next, callus induction were done in two complemented MS medium for 6 weeks. Then, callus propagation were performed in MS medium supplemented with 1.0 mg/L of benzylaminopurine (BAP) and 0.5 mg/L of 2,4-dichlorophenoxyacetic acid (2,4-D) for 50 days. Fresh callus were extracted every 10 days in an ultrasonic bath using ethyl acetate (1.0 g/10 mL). The identification was carried out by Gas Chromatography-Mass Spectrometry (GC-MS) using selected ion monitoring (SIM) acquisition mode with characteristic ions of lupeol. The results obtained indicate the occurrence of lupeol in callus extract after twenty days of proliferation. These findings could be use in subsequent scale-up studies for biomass production containing this active compound in order to replace conventional methods.

Antioxidant activity optimization and GC-MS profile of aqueous extracts of Vernonanthura patens (Kunth) H. Rob. leaves.

The temperature and extraction time of aqueous extracts of Vernonanthura patens (Kunth) H. Rob. (AEVP) leaves obtained by decoction were optimized for maximum recovery of DPPH radical scavenging activity, ABTS+ inhibition activity, total phenolic content (TPC) and total flavonoids content (TFC) using response surface methodology (RSM). A central composite design (CCD) of 13 experimental runs was applied and second order polynomial models were used to describe the responses of the assessed extraction parameters. The optimized conditions: 79.79 °C and 126.23 minutes were found using the composite desirability function. The scavenging activity of assessed extracts could be correlated mostly to the presence of malic acid, succinic acid, α-ketoglutaric acid, citric acid, m-hydroxybenzoic acid, caffeic acid, inositol, and ß-amyrin detected by gas chromatography-mass spectrometry (GC-MS). These results have not been reported and support the potential application of AEVP as natural source of antioxidants.

Effect of resin content and pressing temperature on banana pseudo-stem particle boards properties using full factorial design.

The volume of global gross banana exports reached a record of 117.9 million tonnes in 2015 (FAO 2017), which agro-industrial wastes derived as the pseudo-stem, rachis and leaves do not have an industrial application instead they are discarded. This research study applies full factorial design and response surface methodology to determine the effect of pressing temperature and resin content on density (D), moisture (M), water absorption (WA), water swelling (WS), module of rupture (MOR), module of elasticity (MOE) and formaldehyde content (FC) of particle board made of banana pseudo-stem. A 22 factorial design was performed, factors considered were resin and temperature. The low level of resin was 15% in the coarse fiber (CF) and 35% in fine fiber (FF); high level as 25% CF and 45% FF. Temperature levels were 150ºC and 170ºC respectively. The boards met all quality parameters except ones with low resin content that didn't meet WS parameter. Furthermore, resin affected positively on WA, FC and MOE, and decreased D, WS and MOR values. Meanwhile, temperature affected negatively on D, WS, and increased FC, WA, MOE, MOR properties; none of the factors affected M response. Process conditions were optimized to 162.61°C and 43.15% FF, 23.97% CF.