Anthocyanins Formulated with Carboxymethyl Starch for Gastric and Intestinal Delivery.

Anthocyanins obtained from jambolan have been used as active agents in different carboxymethyl starch-based tablet formulations and their release profiles evaluated in simulated gastric fluids (SGF) and simulated intestinal (SIF) fluids. Structural analysis highlighted a strong interaction between anthocyanins and carboxymethyl starch, evidenced by scanning electron microscopy and infrared analysis. Tablet dissolution behavior varied according to the pH of the media, being controlled by the swelling and/or erosion of the polymeric matrix. Various formulations for immediate, fast, and sustained release of anthocyanins for 30 min, 2 h and 12 h of dissolution have been developed. It was found that monolithic carboxymethyl starch tablets loaded with powdered jambolan extract efficiently afforded the complete delivery (100% of anthocyanins) to different sites of the simulated gastrointestinal tract and ensured the stability of these pigments, which maintained their antioxidant activity.

UPLC-MS-ESI-QTOF Analysis and Antifungal Activity of Aqueous Extracts of Spondias tuberosa.

This study aimed to identify the chemical composition of the Spondias tuberosa aqueous leaf and root extracts (EALST and EARST) and to evaluate their effect, comparatively, against opportunistic pathogenic fungi. Ultra-Performance Liquid Chromatography Coupled to a Quadrupole/Time of Flight System (UPLC-MS-ESI-QTOF) was employed for chemical analysis. Candida albicans and C. tropicalis standard strains and clinical isolates were used (CA INCQS 40006, CT INCQS 40042, CA URM 5974, and CT URM 4262). The 50% Inhibitory Concentration for the fungal population (IC50) was determined for both the intrinsic action of the extracts and the extract/fluconazole (FCZ) associations. The determination of the Minimum Fungicidal Concentration (MFC) and the verification of effects over fungal morphological transitions were performed by subculture in Petri dishes and humid chambers, respectively, both based on micro-dilution. UPLC-MS-ESI-QTOF analysis revealed the presence of phenolic and flavonoid compounds. The association of the extracts with fluconazole, resulted in IC50 values from 2.62 µg/mL to 308.96 µg/mL. The MFC of the extracts was ≥16,384 µg/mL for all tested strains, while fluconazole obtained an MFC of 8192 µg/mL against C. albicans strains. A reduction in MFC against CA URM 5974 (EALST: 2048 µg/mL and EARST: 1024 µg/mL) occurred in the extract/fluconazole association.

Ultrasound-assisted extraction of anthocyanins and phenolics from jabuticaba (Myrciaria cauliflora) peel: kinetics and mathematical modeling.

Jabuticaba (Myrciaria cauliflora) is a dark berry, endemic to the south and central regions of South America, rich in anthocyanins and polyphenols. This study evaluated the ultrasound-assisted extraction of bioactive compounds from jabuticaba peel, developed a new mathematical model for the process, and estimated the model parameters. Extraction was carried out using water as solvent aiming its direct use in food formulations. The main anthocyanin (cyanidin-3-O-glucoside) and the main polyphenol (ellagic acid) from jabuticaba peel were extracted and quantified by LC-MS and HPLC. The results indicate that lowering the pH increased the extraction of the anthocyanin and had only slight effect on the extraction of ellagic acid. The application of ultrasound at 25 kHz favored the extraction of both compounds. Processing time of 20 min increased the yield of both compounds, while over processing (> 20 min) let to the sonochemical-induced hydrolysis of cyanidin-3-O-glucoside and ellagic acid. The highest yield of bioactive compounds was attained at 25 kHz, 20 min of extraction and pH 1.5 (8.9 mg/g dry peel of gallic acid equivalent, 0.9 mg/g dry peel of ellagic acid, and 7.9 mg/g dry peel of cyanidin-3-O-glucoside). The new mathematical model considered the mass transfer between the powder and the liquid media, and the sonochemical-induced hydrolysis of the compound. The model was able to predict satisfactorily the extraction process and the hydrolysis effect.

Simultaneous quantification of lipopeptide isoforms by UPLC-MS in the fermentation broth from Bacillus subtilis CNPMS22.

The rapid and accurate quantification of lipopeptide families in biological samples are challenging. We present the development and validation of a method for simultaneous quantification of three families of lipopeptides (iturins, fengycins, and surfactins) and their isoforms, as well as the homologous series. The method was optimized in UPLC-MS for a column temperature at 65 °C, injection volume of 5 µL, and sample temperature of 10 °C. The SIM mode was used for detection and quantification of lipopeptides exhibiting ions [M + H]+ and [M + 2H]2+. Since the maximum mass detection threshold of the equipment is 1250 Da and the fengycins have ions between 1435 and 1505 Da, the ions [M + 2H]2+ were chosen for fengycin identification. The monitored ions were as follows: m/z 1043.5, 1057.5, 1071.5, 718.3, 725.4, 739.4, 732.4, 746.4, 753.4, 1008.6, 1022.6, and 1036.6. The compounds were separated by reverse-phase chromatography using a C18 analytical column in a total time of 19 min. Standard curves were linear with rw 0.99 for all analytes. Intra- and inter-day precision for samples (50, 250, and 750 µg L-1) were within recommended limits. The proposed analytical method was capable of simultaneously quantifying 12 isoforms and homologous series of lipopeptide families in biological samples, thus making it an important industrial tool in the evaluation of lipopeptide production processes. Graphical abstract ᅟ.

Effect of freeze- and spray-drying on physico-chemical characteristics, phenolic compounds and antioxidant activity of papaya pulp.

The effect of freeze- and spray-drying on physico-chemical characteristics, phenolics compounds and antioxidant activity of papaya pulp was investigated. The frozen pulp was freeze dried at - 62 °C during 48 h. Papaya pulp with 14% DE maltodextrin was also dried in a spray dryer. The organic acids, sugars, color, total soluble solids, pH, carotenoids, phenolic and flavonoid compounds, and antioxidant capacity values were determined. The changes in color, pH and lycopene were negligible. However, lower retention (86.5%) of vitamin C and sugars (glucose-79.7% and fructose-66.1%) was observed in spray dried products. Phenolic and flavonoid compounds were identified and quantified in dried papaya products by UHPLC-QqQ-MS/MS. Spray dried products presented a higher retention of phenolic and flavonoid compounds compared to the freeze dried products. Despite some variations in the parameters studied, the use of freeze- and spray-drying has proven viable options for the drying of papaya pulp.

Influence of pectinolyttic and cellulotyc enzyme complexes on cashew bagasse maceration in order to obtain carotenoids.

Cashew apple bagasse is a byproduct of cashew peduncle juice processing. Such waste is a source of carotenoids, but it is usually discarded after the juice extraction. The objective of this work was to study the influence of pectinolytic and cellulolytic enzyme complexes on cashew bagasse maceration in order to obtain carotenoids. It was observed that maceration with the enzymatic complex Pectinex Batch AR showed a higher content of carotenoids, with an overall gain of 79 % over the control carried out without enzyme complex addition.

Comparison of α and ß-acid isomerization in hops and beer using HPLC, confocal microscopy, spectrofluorimetry and chemical analysis of metabolites and essential oils in flowers of different hop cultivars produced in Brazil.

We used confocal microscopy and spectrofluorescence to characterize the emission spectra in hop flowers, to follow the isomerization processes in different hop preparations, and beers, to compare with HPLC extracted samples. Flowers of different hop cultivars produced in three regions of Brazil, were quantitated by HPLC and GC-MS. The fluorescence spectra showed two characteristic emission bands evaluated from different preparations. The isomerization process leads to a gradual decrease in fluorescence intensity as the reaction progresses. This demonstrates the valuable use of confocal microscopy and fluorescence spectroscopy for analysis of the correlation between bitter acid indices with fluorescence intensity and lifetime microscopy. Such techniques can be used directly in the flowers allowing rapid monitoring of the brewing process. Twenty-nine substances were characterized in the essential oils and some cultivars presented quantities of bitter acids and essential oil levels close to those expected for plants after more than three years of cultivation.

Validation of a Method for Anacardic Acid Quantification in Cashew Peduncles via High-Performance Liquid Chromatography Coupled to a Diode-Array Detector.

The cashew peduncle has a high nutritional value and contains a wide variety of phenolic compounds. Among these, anacardic acids (AnAc) are biologically active components; however, they influence the cashew juice flavor and, consequently, its acceptance. This study validates a high-performance liquid chromatography method for quantifying the AnAc present in cashew peduncles, using a C18 reverse-phase column and a diode-array detector. The calibration curve obtained showed satisfactory precision for intraday (CV = 0.20%) and interday (CV = 0.29%) quantification, linearity (y = 2333.5x + 2956.2; r2 = 0.9979), repeatability with respect to retention time (CV = 0.45%) and area (CV = 0.30%), and selectivity, and possessed detection and quantification limits of 0.18 and 0.85 µg·mL-1, respectively. Different cashew clones containing AnAc were extracted and analyzed using the proposed method. A recovery of >90% was achieved using two sequential extractions. The total AnAc content ranged from 128.35 to 217.00 mg·100 g-1 in peduncle samples obtained from five different cashew clones.

Effect of Ball-Milling on Starch Crystalline Structure, Gelatinization Temperature, and Rheological Properties: Towards Enhanced Utilization in Thermosensitive Systems.

Starch's crystalline structure and gelatinization temperature might facilitate or hinder its use. Ball milling has frequently been mentioned in the literature as a method for reducing starch size and as a more environmentally friendly way to change starch, such as by increasing surface area and reactivity, which has an impact on other starch properties. In this study, starch samples were milled for varying durations (1, 5, 10, 20, and 30 h) and at different starch-to-ball mass ratios (1:6 and 1:20). Microscopy and XRD revealed that prolonged milling resulted in effective fragmentation and a decrease in crystallinity of the starch granules. Increasing milling times resulted in an increase in amylose content. Rheology and thermal studies revealed that gelatinization temperatures dropped with milling duration and that viscosity and thixotropy were directly influenced. The samples milled for 10, 20, and 30 h at a ratio of 1:20 were the most fragmented and upon drying formed a transparent film at ambient temperature, because of the lower gelatinization temperature. Starch ball milling could lead to the use of this material in thermosensitive systems.

UPLC-QTOF-MS/MS-based metabolomic approach and gastroprotective effect of two chemotypes of Egletes viscosa (L.) less. against ethanol-induced gastric ulcer in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Egletes viscosa (L.) (macela) is a native wild herb that can be found in different states of northeastern Brazil. The infusions of its flower buds are traditionally used for the treatment of gastrointestinal disorders. E. viscosa possesses two chemotypes (named A and B), distinguishable by the composition of the essential oil from the flower buds. Although there are previous studies of the gastroprotective effect of the isolated constituents of E. viscosa, its infusions have not been investigated yet. AIM OF THE STUDY: The present study aimed to evaluate and compare the chemical composition and the gastroprotective effect of flower bud infusions of E. viscosa from chemotype A (EVCA) and chemotype B (EVCB). MATERIALS AND METHODS: Sixteen infusions were brewed with flower buds according to the traditional preparation mode and were analyzed through a UPLC-QTOF-MS/MS based metabolomic approach for determination of their metabolic fingerprints and quantification of bioactive compounds. Afterward, these data were analyzed by chemometric methods (OPLS-DA) for discrimination of the two chemotypes. Additionally, infusions of EVCA and EVCB (50, 100 and 200 mg/kg, p.o.) were evaluated on gastric ulcers induced by absolute ethanol (96%, 0.2 mL, p.o.) in mice. To elucidate the gastroprotective mechanisms, the effect of EVCA and EVCB on gastric acid secretion and gastric wall mucus was determined and the role of TRPV1 channels, prostaglandins, nitric oxide and KATP channels were assessed. Moreover, the oxidative stress-related parameters and the histological aspects of the stomach tissue were analyzed. RESULTS: The chemotypes can be discriminated from each other using UPLC-QTOF-MS/MS chemical fingerprints. Both chemotypes presented similar chemical compositions, consisting basically of caffeic acid derivatives, flavonoids and diterpenes. The quantification of bioactive compounds demonstrated that chemotype A possesses more ternatin, tanabalin and centipedic than chemotype B. EVCA and EVCB (50, 100 and 200 mg/kg, p.o.) significantly decreased the severity of ethanol-induced gastric lesions, as shown by a reduction in histological alterations and leucocyte infiltration in gastric tissue. The gastroprotective mechanism of both infusions involves an antioxidant effect, maintenance of gastric mucus and reduction gastric secretion. Stimulation of endogenous prostaglandins and nitric oxide release, activation of TRPV1 channels, and KATP channels are also involved in the gastroprotection of the infusions. CONCLUSION: The gastroprotective effect of EVCA and EVCB was equivalent and mediated through antioxidant and antisecretory actions, including the activation of TRPV1 receptors, stimulation of endogenous prostaglandins and nitric oxide, and opening of KATP channels. The presence of caffeic acid derivatives, flavonoids and diterpenes in both infusions is involved in mediating this protective effect. Our findings support the traditional use of infusions of E. viscosa for gastric disorders regardless of the chemotype.

LC-MS Analysis and Antifungal Activity of Turnera subulata Sm.

Fungi of the Candida genus are responsible for invasive candidiasis, which affects people all over the world and has high mortality rates. This is due to their virulence factors, which give them great resistance and pathogenicity. In addition, the emergence of multidrug-resistant strains makes it difficult to treat these infections. In this way, natural products have emerged as an alternative to standard drugs, where plants known for their medicinal properties such as Turnera subulata become attractive to research. The present work aimed to analyze the ethanol extract of Turnera subulata leaves against standard strains of Candida albicans, Candida krusei and Candida tropicalis using broth microdilution techniques. The identification of the compounds in T. subulata leaves by LC-MS revealed the presence of a wide variety of substances such as carboxylic acids and terpenes, with flavonoids and fatty acids being more evident. The antifungal assays showed that the extract was not able to inhibit the growth of the tested strains at concentrations with a clinical relevance. However, at higher concentrations, it was able to inhibit the fungal dimorphism of C. albicans and C. tropicalis. It is possible that the T. subulata extract has potential as an inhibitor of fungal virulence factors without affecting the cell viability. Further research should be carried out in order to assess its inhibitory potential for other fungal virulence factors.

Untargeted metabolomic profile of recovered bioactive compounds by subcritical water extraction of acerola (Malpighia emarginata DC.) pomace.

The untargeted metabolomics approach was used to compare the chemical profiles of acerola (Malpighia emarginata DC.) pomace extracts. The effect of drying the raw material before subcritical water extraction (SWE) at different temperatures on the yield, phenolic content, and in vitro antioxidant activity was evaluated. The results were compared with those obtained via Soxhlet and the findings suggest that SWE saves time (15 min) and solvent for extracting valuable components as compared to Soxhlet (6 h). An increase in temperature significantly improved the extraction yield (23.9 to 33.4 %), phenolic content (119.1 to 362 mgGAEg-1), and antioxidant activity, and higher values were obtained with SWE as compared to Soxhlet. The most abundant compounds detected by UPLC-ESI-QTOF-MS were ascorbic acid, kaempferol, quercetin, and isorhamnetin. The investigation of different moisture contents in the SWE showed promising results for eliminating the drying operation, saving time and energy, and obtaining highly concentrated phenolic-rich by-products.

Anti-Candida Properties of Gossypium hirsutum L.: Enhancement of Fungal Growth, Biofilm Production and Antifungal Resistance.

(1) Background: Candida is a genus of yeasts with notable pathogenicity and significant ability to develop antimicrobial resistance. Gossypium hirsutum L., a medicinal plant that is traditionally used due to its antimicrobial properties, has demonstrated significant antifungal activity. Therefore, this study investigated the chemical composition and anti-Candida effects of aqueous (AELG) and hydroethanolic (HELG) extracts obtained from the leaves of this plant. (2) Methods: The extracts were chemically characterized by UPLC-QTOF-MS/MS, and their anti-Candida activities were investigated by analyzing cell viability, biofilm production, morphological transition, and enhancement of antifungal resistance. (3) Results: The UPLC-QTOF-MS/MS analysis revealed the presence of twenty-one compounds in both AELG and HELG, highlighting the predominance of flavonoids. The combination of the extracts with fluconazole significantly reduced its IC50 values against Candida albicans INCQS 40006, Candida tropicalis INCQS 40042, and C. tropicalis URM 4262 strains, indicating enhanced antifungal activity. About biofilm production, significant inhibition was observed only for the AELG-treated C. tropicalis URM 4262 strain in comparison with the untreated control. Accordingly, this extract showed more significant inhibitory effects on the morphological transition of the INCQS 40006 and URM 4387 strains of C. albicans (4) Conclusions: Gossypium hirsutum L. presents promising antifungal effects, that may be potentially linked to the combined activity of chemical constituents identified in its extracts.

Chemical profile of colorful bean (Phaseolus vulgaris L) flours: Changes influenced by the cooking method.

This study aimed to determine how the cooking methods change the phenolics and saponins profiles, oligosaccharides, antinutrients and antioxidant properties of flours from colorful beans. The autoclave cooking consisted of: 6 h soaking and 5 min cooking (C5); and 20 min cooking without soaking (C20). Both cooking methods significantly promote changes on the chemical compounds studied, and the intensity of these variations were affected by the cultivars. Most of flours of C5 beans presented a lower loss of anthocyanins (3.9-70.0%), DPPH (11.7-87.2%), ABTS (0.0-82.7%), and tannins (0.0-90.0%) compared with C20. The cooked flours of Artico and Realce showed some similarities among chemical compounds, as well as the lowest concentration of tannins (0.0 mg‧g-1), antioxidant activity (0.40 µmol Trolox‧g-1), and higher amounts of oligosaccharides and acetylcholine. Most of cooked flours presented a reduction in phenolics and soyasaponins αg and ßg, and an increase in soyasaponins Ba and I and oligosaccharides (mainly C20 flours).

Corn starch based films treated by dielectric barrier discharge plasma.

Cold plasma is an innovative strategy to strengthen the polysaccharide-based films characteristics. This study evaluated the effects of dielectric barrier discharge (DBD) plasma on the hydrophilic character, water vapor permeability (WVP), and tensile properties of corn starch-based films. Starch films were exposed to plasma processing operating at an excitation frequency of 200 Hz for 10, 15, and 20 min. DBD plasma resulted in further enhanced tensile strength and stiffness, and lower hydrophilicity and water solubility; however, it did not present significant effects on the WVP of the resulting films within the ranges studied. Higher hydrophobicity, strength, and stiffness were verified after 20 min. The results presented in this work suggest that the DBD plasma has the potential to make starch-based films a more suitable packaging material.

Characterization of Anti-Ana o 3 Monoclonal Antibodies and Their Application in Comparing Brazilian Cashew Cultivars.

Ana o 3 is an immuno-dominant cashew nut allergen. Four monoclonal antibodies to Ana o 3 (2H5, 6B9C1, 19C9A2, and 5B7F8) were characterized by ELISA and in silico modeling. The 2H5 antibody was the only antibody specific for cashew nut extract. In addition to cashew nut extract, the 6B9C1 and 19C9A2 antibodies recognized pistachio extract, and the 5B7F8 recognized pecan extract. All four antibodies recognized both recombinant Ana o 3.0101 and native Ana o 3. ELISA assays following treatment of purified Ana o 3 with a reducing agent indicated that the 6B9C1 and 19C9A2 antibodies likely recognize conformational epitopes, while the 2H5 and 5B7F8 antibodies likely recognize linear epitopes. In silico modeling predicted distinct epitopes for each of the anti-Ana o 3 antibodies. Screening extracts from 11 Brazilian cashew nut cultivars using all four antibodies showed slight differences in Ana o 3 bindings, demonstrating that these antibodies could identify cultivars with varying allergen content.

Antioxidant Activity of Stryphnodendron rotundifolium Mart. Stem Bark Fraction in an Iron Overload Model.

Stryphnodendron rotundifolium Mart., popularly known as "barbatimão", is a plant species traditionally used by topical and oral routes for the treatment of infectious and inflammatory diseases. Considering the well-described antioxidant properties of this species, this study investigated the protective effects of its keto-aqueous extract using an in vitro model of iron overload. Phenolic compounds were quantified and identified by Ultra-Performance Liquid Chromatography coupled with quadrupole Time-Of-Flight Electrospray Ionization Mass Spectrometry (UPLC-ESI-qTOF-MS/MS) in positive and negative ions mode analysis. Antioxidant activity was analyzed following the iron-chelating-reducing capacity and deoxyribose degradation (2-DR) protection methods. The analysis identified condensed tannins (54.8 mg catechin/g dry fraction (DF), polyphenols (25 mg gallic acid/g DF), and hydrolyzable tannins (28.8 mg tannic acid/g DF). Among the constituents, prodelphinidin, procyanidin, and prorobinetinidine were isolated and identified. The extract significantly protected 2-DR degradation induced by Fe2+ (72% protection) or •OH (43% protection). The ortho-phenanthroline test revealed Fe2+-chelating and Fe3+-reducing activities of 93% and 84%, respectively. A preliminary toxicological analysis using Artemia salina revealed mortality below 10%, at a concentration of 0.25 mg/mL, indicating low toxicity under the present experimental conditions. In conclusion, the findings of the present study indicate that Stryphnodendron rotundifolium is a source of antioxidant compounds with the potential to be used in drug development in the context of iron overload disorders, which remains to be further investigated in vivo.

From mango by-product to food packaging: Pectin-phenolic antioxidant films from mango peels.

The objective of the study was to prepare active films based on pectin and polyphenol-rich extracts from Tommy Atkins mango peels. Aqueous and methanolic extracts showed a variety of phenolic compounds that were identified by UPLC-MS analysis, and a high content of total phenolics that were quantified by the Folin-Ciocalteau method. The methanolic extract showed better results in antioxidant tests and was more effective in inhibiting the growth of Gram-positive and Gram-negative bacteria. The pectin extracted from mango peels showed good thermal stability and a degree of methoxylation of 58.3% by 1H NMR. The films containing the phenolic extracts showed lower water vapor permeability when compared to the control film (without any phenolic extracts). The incorporation of the extracts led to an increase in elongation (ε) and a decrease in tensile strength (σ) and modulus of elasticity (Y). The films with aqueous or methanolic extracts showed higher antioxidant activity in terms of inhibition of the DPPH radical. Therefore, the films developed in this work are presented as a promising alternative for food packaging and/or coating applications.

Ozone and plasma processing effect on green coconut water.

In this study, the effect of plasma and ozone processing on the quality of coconut water was evaluated. For ozone processing, the samples were submitted to different ozone loads and temperatures. For atmospheric cold plasma processing (ACP), samples were exposed to plasma under different frequencies and voltages. The coconut water pH, soluble solids, titratable acidity, color, total phenolic content, and enzymatic activity were determined before and after treatments. The main compounds were also determined by NMR spectroscopy and chemometric analysis. Both processes did not change the pH values, total soluble solids, titratable acidity, and color. Chemometrics analysis of 1H NMR dataset showed no relevant changes after the processing. All ozone treatments promoted complete inactivation of POD activity and did not affect the content of phenolic compounds. After ACP, the smallest POD residual activity was observed when higher frequencies were applied, and slight changes in phenolic compounds content were observed.

UPLC-MS-QTOF analysis and antifungal activity of Cumaru (Amburana cearensis).

This study was aimed at investigating the phytochemical constituents, antifungal properties and antibiotic-modifying activity of the aqueous crude extract and fractions of Amburana cearensis seeds (CEFAC). The CEFAC were chemically characterized by LC-MS/MS-QTOF. In addition, the antifungal activity was assayed by the microdilution method against strains of Candida albicans. The phytochemical profile of CEFAC exhibited phenolic compounds, organic acids, and polyphenols. The results of the assessment of antifungal activity reveled an IC50 ranging from 45.6 to 2048 µg/mL. Interestingly, when CEFAC was associated with Fluconazole, we evidenced a decreased IC50 (1.81-11.9 µg/mL), suggesting a synergism with antibiotic. It was possible to identify in the crude extract and fractions several phenolic compounds, organic acids, and some polyphenols in positive ionization mode. These results suggest that CEFAC may present compounds with the ability to interact and act synergistically with antimicrobial drugs, highlighting its potential as an alternative source for the development of new antimicrobial agents.