Metabolomic Profiling of Phloem Sap from Different Pine Species and Implications on Black Capuchin.

In most commercial pine farms in southern Brazil, black capuchin causes damage to wood and financial losses when it removes bark from some pine species to feed upon underlying vascular tissues. Therefore, this study aimed to evaluate the variability of the primary metabolites of phloem saps from 10 different species of pine by NMR spectroscopy, as well as the aroma compounds using SPME-GC-MS. Each technique provided a different set of metabolites that we can correlate to monkey predilection. The PCA showed monosaccharide (detected by NMR) and α-pinene (pine-like and resinous flavor descriptors) as attractive compounds for monkeys. On the other hand, the low content of monosaccharide and the high content of ß-phellandrene (citrus odor descriptor) was observed in less attacked pine species (P. patula). The data fusion on primary metabolites and aroma compounds corroborated the individual analyses, complementing the comprehension of the monkey predilection. Thus, P. elliottii was an avoided tree even with high content of sugars possibly due to its high content of ß-phellandrene (citrus odor). The results are useful for further behavioral studies to determine the role that each highlighted metabolite plays in chemically mediated animal-plant interactions.

Association of Pollinators of Different Species of Oil Palm with the Metabolic Profiling of Volatile Organic Compounds.

The development of studies on emissions of volatile organic compounds (VOCs) by inflorescence of oil palms deserves a special attention regarding the importance to reproduction success and for increase of production. This study aimed to evaluate metabolic profiling of VOCs expelled by male and female inflorescences of different oil palm species (African oil palm, Amazonian Caiaué and the interspecific hybrid BRS-Manicoré), associating the composition variability with main pollinators to improve the comprehension of the plant-insect relationship. The phenylpropanoids, terpenoids and the aliphatic hydrocarbons were predominant classes detected in inflorescences of oil palms and the major compound was estragole. This result may be correlated with attraction of Elaidobius pollinators, since these insects were not attracted by Caiaué, which emitted estragole only in trace amounts. However, Caiaué and the hybrid species were visited by other native species whose frequencies were low and their success as pollinators could not be expected.

Sequential extraction of flavonoids and pectin from yellow passion fruit rind using pressurized solvent or ultrasound.

BACKGROUND: Passion fruit rind (PFR) represents 90% of the total fruit weight and is wasted during juice processing. Passion fruit rind is known to contain flavonoids and pectin. An alternative use for this fruit juice industrial residue is to obtain these compounds. This study aimed to verify the influence of pressurized solvent extraction (PSE) or ultrasound assisted extraction (UAE) of flavonoid and pectin in a sequential process. RESULTS: The PSE using ethanol at 60:40 (v/v) yielded a total polyphenol content of 4.67 g GAE kg-1 PFR, orientin-7-O-glucoside (1.57 g kg-1 PFR) and luteolin-6-C-glucoside (2.44 g kg-1 PFR). Pectin yield was 165 g kg-1 PFR, either in PSE or UAE. Pectin characterization indicates that the pectic structure has basically homogalacturonans and galacturonate followed by a galacturonic acid ester unit, with methylation degree of 70%. CONCLUSION: With this study it can be concluded that mixtures of alcohols with water favor the extraction of bioactive compounds of passion fruit peel. Both PSE and UAE were effective in sequentially extracting flavonoids and pectin. The preferred solvent is ethanol due to its lower toxicity. © 2017 Society of Chemical Industry.

Synthesis, antibacterial and cytotoxic activities of new biflorin-based hydrazones and oximes.

Biflorin 1 is a biologically active quinone, isolated from Capraria biflora. Five new biflorin-based nitrogen derivatives were synthesized, of which two were mixtures of (E)- and (Z)- isomers: (Z)-2a, (Z)-2b, (Z)-3a, (Z)- and (E)-3b, (Z)- and (E)-3c. The antibacterial activity was investigated using the microdilution method for determining the minimum inhibitory concentration (MIC) against six bacterial strains. Tests have shown that these derivatives have potential against all bacterial strains. The cytotoxic activity was also evaluated against three strains of cancer cells, but none of the derivatives showed activity.

Chilled Pacu (Piaractus mesopotamicus) fillets: Modeling Pseudomonas spp. and psychrotrophic bacteria growth and monitoring spoilage indicators by 1H NMR and GC-MS during storage.

This study aimed to assess the growth of Pseudomonas spp. and psychrotrophic bacteria in chilled Pacu (Piaractus mesopotamicus), a native South American fish, stored under chilling conditions (0 to 10 °C) through the use of predictive models under isothermal and non-isothermal conditions. Growth kinetic parameters, maximum growth rate (µmax, 1/h), lag time (tLag, h), and (Nmax, Log10 CFU/g) were estimated using the Baranyi and Roberts microbial growth model. Both kinetic parameters, growth rate and lag time, were significantly influenced by temperature (P < 0.05). The square root secondary model was used to describe the bacteria growth as a function of temperature. Secondary models, √µ = 0.016 (T + 10.13) and √µ =0.017 (T + 9.91) presented a linear correlation with R2 values >0.97 and were further validated under non-isothermal conditions. The model's performance was considered acceptable to predict the growth of Pseudomonas spp. and psychrotrophic bacteria in refrigerated Pacu fillets with bias and accuracy factors between 1.24 and 1.49 (fail-safe) and 1.45-1.49, respectively. Fish biomarkers and spoilage indicators were assessed during storage at 0, 4, and 10 °C. Volatile organic compounds, VOCs (1-hexanol, nonanal, octenol, and indicators 2-ethyl-1-hexanol) showed different behavior with storage time (P > 0.05). 1H NMR analysis confirmed increased enzymatic and microbial activity in Pacu fillets stored at 10 °C compared to 0 °C. The developed and validated models obtained in this study can be used as a tool for decision-making on the shelf-life and quality of refrigerated Pacu fillets stored under dynamic conditions from 0 to 10 °C.

NMR-Based Metabolomic Approach for Evaluation of the Harvesting Time and Cooking Characteristics of Different Cassava Genotypes.

Cassava is an important staple food for low-income countries. However, its cooking characteristics are especially affected by genotype. In this study, two groups of genotypes, namely hard to cook (HTC) and easy to cook (ETC), were harvested at different times (9 and 15 months), and evaluated by NMR coupled to chemometrics. Additionally, lignin of these materials was studied by 1H-13C HSQC NMR. The carbohydrates were the most important class of compounds to differentiate the cassava genotypes. The correlation of NMR with cooking time and starch content showed that the higher content of primary metabolites, mostly glucose, can be associated with longer cooking times and reduction of starch, corroborating the metabolic pathways analysis. Furthermore, it was observed that the lignin from cell walls did not differentiate the cooking performance of the genotypes.

Green ultrasound-assisted extraction of chlorogenic acids from sweet potato peels and sonochemical hydrolysis of caffeoylquinic acids derivatives.

Sweet potato peels are rich in chlorogenic acids. In this work, we applied ultrasound technology to extract the main compounds from sweet potato peel and used multivariate analysis and principal component analysis (PCA) to evaluate the effects of different extraction conditions on the extraction of chlorogenic acids. The extraction was studied varying ultrasonic power density (20, 35 and 50 W/L) and processing time (5, 10, 20 and 40 min) using an ultrasonic bath operating at 25 kHz. The chemical analysis was carried out by UPLC-qTOF-MS, and the results were evaluated by PCA and PLS-DA chemometric analysis. Results show that both ultrasonic power density and processing time influences in the extraction of different chlorogenic acid, and that different extraction conditions can be used to selectively extract specific caffeoylquinic acids and feruloylquinic acids in higher amounts. Ultrasound promoted the hydrolysis of tricaffeoylquinic acid when subjected to ultrasonic waves (20-50 W/L), and of 3,4-caffeyolquinic acid at high ultrasonic power density (50 W/L).

Thermal and non-thermal processing effect on açai juice composition.

This study evaluated the effects of High-Temperature Short Time (HTST), Ultra High Temperature (UHT), and the non-thermal processes High Power Ultrasound (US), UV-pulsed-light and Low Pressure Plasma (LPP) on the composition, stability, and bioactive compounds bioaccessibility of açai juice. 1H NMR based approach, coupled to chemometrics, was applied to evaluate the changes in the juice composition. All the non-thermal processes increased the sugars content (glucose and fructose), and the amino acid betaine, except the combined processing of ultrasound followed by low-pressure plasma (US.LPP). HTST and UHT increased the fatty acids and phenolic compounds content in the açai juice. The bioaccessibility of phenolic compounds decreased due to the processing. After thermal sterilization (UHT), the anthocyanin bioaccessibility was 2-fold higher. The combined non-thermal treatment reduced the biocompounds bioaccessibility to 40% of the non-processed juice. However, the combined US.LPP improved the bioaccessibility of vitamin C by 8%. UHT increased the anthocyanin's bioaccessibility but sharply decreased vitamin C bioaccessibility. Higher impact of thermal processing on vitamin C, anthocyanins, total phenolics, PPO, POD, DPPH, ABTS, and FRAP was verified after 45 and 60 days of storage compared to the non-thermally processed samples.

Pigment production by Fusarium solani BRM054066 and determination of antioxidant and anti-inflammatory properties.

The fungal kingdom has been widely studied as a source of bioactive compounds of interest to the pharmaceutical and food industry. This paper studies the production of natural red pigments by Fusarium solani BRM054066 in the submerged fermentation system, using Doehlert experimental design to determine optimal cultivation conditions. The chemical composition of the red pigment was determined by Nuclear Magnetic Resonance spectroscopy (NMR) and Ultra-Performance Liquid Chromatography coupled to Mass Spectrometry (UPLC-MS). Antioxidant activity was assessed by the ability to sequester of free radical DPPH. In the analysis of anti-inflammatory activity, murine peritoneal macrophages activated by LPS were used, and the gene expression of TNF-α, IL-1ß, IL-6, IL-10 and IL-17 was determined using qPCR. As a result, it was found that agitation at 200 rpm and glucose concentration ≥ 20 g/L promote the best results in the production of red pigment. The chemical compounds identified were two naphthoquinones, fusarubin and dihydrofusarubin, and an anthraquinone, a bostrycoidin, being fusarubin the majority compound. The red pigment showed antioxidant activity by scavenge 50% of the DPPH radical, in a concentration of 24 µg/mL. The pigment also showed an effective anti-inflammatory capacity by reducing the overexpression of the pro-inflammatory cytokines TNF-α, IL-1ß and IL-6 and promoting the production of anti-inflammatory IL-10 and IL-17, in murine macrophages activated by LPS (p < 0.05). According to the results, the fungus F. solani BRM054066, under optimized conditions of cultivation, proved to be a promising source of biologically active natural pigments with wide industrial applicability.

An integrated analytical approach based on NMR, LC-MS and GC-MS to evaluate thermal and non-thermal processing of cashew apple juice.

Innovative chemometric approaches by NMR and LC-MS data fusion (multiblock analysis) and decomposition of the GC-MS raw data by PARADISe were applied to evaluate the influence of thermal and non-thermal processing on the composition of cashew apple juices. Comparative investigations by Principal Component Analysis (PCA) highlighted pronounced effect of thermal processing on juice compared to non-thermal processing, as decreases of anacardic acids, sucrose, malic acid, tyrosine, phenylalanine, and important flavor compounds (esters, aldehydes, and ketones). Ultrasound presented relevant influence on increase of anacardic acids concentration. Non-thermal processing carried out at more intense conditions (10 min of ultrasound, 5 min of ozone, and pulsed light at 10 V) showed pronounced effect compared to other non-thermal processing. Although individual PCA enables to detect the influence of different processing technologies, data fusion and PARADISe presented advantages, since a more comprehensive understanding of the relationship among chemical changes from different analytical techniques were established.

NIR and 1H qNMR methods coupled to chemometrics discriminate the chemotypes of the gastroprotective herb Egletes viscosa.

Egletes viscosa is a Brazilian medicinal herb consumed as flower bud tea due to its gastroprotective properties. This plant possesses two essential oil-based chemical varieties: trans-pinocarveyl acetate-rich chemotype A and cis-isopinocarveyl acetate- rich chemotype B. Therefore, we developed two simple, fast and reliable methods for discrimination of E. viscosa chemotypes using NIR and 1H qNMR spectroscopies combined with the chemometrics tools (iPLS and PLS-DA). Both methods showed high sensitivity, precision and specificity in the cross-validation tests. The NIR method has the advantages of being non-destructive and analyzable by portable devices, enabling its application for field and industrial evaluations. Meanwhile, the 1H qNMR method allows the quantification of the bioactive components ternatin, tanabalin, and centipedic acid. These aforementioned compounds were found higher in the chemotype A. Accordingly, our methods showed to be complimentary approaches for authenticity and/or quality control of E. viscosa-derived raw materials and herbal products.

Cold plasma processing effect on cashew nuts composition and allergenicity.

The study investigated the influence of atmospheric plasma processing on cashew nut composition as well as on its allergenicity. The cashew nuts were processed by low-pressure plasma, using glow discharge plasma (80 W and 50 kHz power supply). Anacardic acids and allergens were quantified by HPLC and immunoassay, respectively. Additionally, the overall composition was evaluated by 1H qNMR. Increases in amounts of anacardic acids (15:1, 15:2, and 15:3) and fatty acids (oleic, linoleic, palmitic and stearic) were detected after all process conditions, with 70.92% of total variance captured using 2 LVs. The total amount of anacardic acids increased from 0.7 to 1.2 µg·mg-1 of nut. The major change was observed for anacardic acid (C15:3) with an increase from 0.2 to 0.55 µg/mg of nut for the samples treated with a flow of 10 mL·min-1 and 30 min of processing. On the other hand, the amount of sucrose decreased, from 33 to 18 mg·g-1 of nut, after all processing conditions. Plasma processing of cashew nuts did not affect binding of either the rabbit anti-cashew or human cashew allergic IgE binding. Among the treatments, 10 min of plasma processing at flow rate of 30 mL·min-1 of synthetic air followed by 20 min at flow rate 5.8 mL·min-1 had the least effect on nut composition as a whole.

1H NMR and LC-MS-based metabolomic approach for evaluation of the seasonality and viticultural practices in wines from São Francisco River Valley, a Brazilian semi-arid region.

São Francisco River Valley (SFRV) is a wine-producing semi-arid region in Brazil. Therefore, we used a 1H NMR and UPLC-MS-based metabolomic approach coupled to chemometrics to evaluate the variability in Chenin Blanc and Syrah wines for two harvest seasons, two vine training system and six rootstocks. Overall, the secondary metabolites were influenced by the three factors studied, whereas the primary metabolites were only by the seasonality. Chenin Blanc wines made in December presented higher content of an unidentified carbohydrate. In Syrah wines, glycerol, tartaric acid, succinic acid and 2,3-butanediol were greater in December, while proline and lactic acid were more abundant in July. For training system, caffeic acid derivatives were increased in wines produced from espalier. Lyre system increased phenolic compounds, organic acids and apocarotenoids. The effect of the rootstocks was less pronounced, affecting basically caffeic acid derivatives. Thus, we expect that our results may assist the winemakers to improve the SFRV wine quality.

Physiological changes for drought resistance in different species of Phyllanthus.

The Phyllanthus genus is widely distributed in tropical and subtropical areas of the world and present several pharmacological applications. Drought is a restrictive factor for crop development and production, and is becoming a severe problem in many regions of the world. The species Phyllanthus amarus and Phyllanthus niruri were subjected to drought stress for varying periods of time (0, 3, 5, 7, and 10 days), and afterwards, leaves were collected and evaluated for physiological and biochemical responses, such as oxidative stress markers and drought-associated defense mechanisms. Results show that P. amarus has an endogenously higher level of variables of the oxidative/antioxidant metabolism, and P. niruri presents the most significant changes in those variables when compared to control and stressed plants. For both Phyllanthus species, drought stress induces higher levels of organic acids such as malic, succinic, and citric acids, and amino acids such as proline, GABA, alanine, and valine. Moreover, P. niruri plants respond with greater glucose and corilagin contents. Therefore, considering the evaluated metabolic changes, P. amarus is better adapted to drought-stress, while P. niruri presents an acclimation strategy that increases the corilagin levels induced by short-term drought stress.

Chemometric analysis of NMR and GC datasets for chemotype characterization of essential oils from different species of Ocimum.

The genus Ocimum (Labiatae) comprises 30 species found in tropical and subtropical regions of the planet, of which species O. basilicum L. and O. gratissimum are widely used in food and traditional medicine. Phytochemical studies on Ocimum have revealed a number of essential oil chemotypes, for example, eugenol, methyl chavicol, linalool, and methyl cinnamate. Since essential oils are commercially assessed according to their content, the aim of this study was to develop a simple and precise method for their qualitative and quantitative analysis using NMR spectroscopy combined with chemometrics. Seven essential oils from different species of Ocimum, an unknown sample, and a commercial sample were evaluated and the results compared to those from established and precise GC-MS and GC-FID methods. Chemometric evaluation from both 1H NMR and GC-MS data revealed three chemotypes: eugenol for O. gratissimum, O. micranthum, and O. tenuiflorum; estragole for O. basilicum, O. basilicum var. purpuracens, and O. selloi; and methyl cinnamate for O. americanum. The unknown and commercial species were classified as cinnamate and eugenol chemotypes, respectively. Despite the corroborating results, the chemometric analysis revealed the higher robustness (better adjustment) of the 1H NMR model compared to the GC-MS method in terms of certain statistical parameters. The 1H NMR method allows for the detection and quantification of organic compounds in a complex mixture without the need for certified standard compounds. Although GC-MS and GC-FID were able to detect five compounds not observed by NMR spectroscopy, the four most important metabolites (eugenol, estragole, methyl cinnamate, and eucalyptol) were more readily detected and quantified by 1H NMR.

Evaluation of thermal and non-thermal processing effect on non-prebiotic and prebiotic acerola juices using 1H qNMR and GC-MS coupled to chemometrics.

The effects of thermal (pasteurization and sterilization) and non-thermal (ultrasound and plasma) processing on the composition of prebiotic and non-prebiotic acerola juices were evaluated using NMR and GC-MS coupled to chemometrics. The increase in the amount of Vitamin C was the main feature observed after thermal processing, followed by malic acid, choline, trigonelline, and acetaldehyde. On the other hand, thermal processing increased the amount of 2-furoic acid, a degradation product from ascorbic acid, as well as influenced the decrease in the amount of esters and alcohols. In general, the non-thermal processing did not present relevant effect on juices composition. The addition of prebiotics (inulin and gluco-oligosaccharides) decreased the effect of processing on juices composition, which suggested a protective effect by microencapsulation. Therefore, chemometric evaluation of the 1H qNMR and GC-MS dataset was suitable to follow changes in acerola juice under different processing.

1H NMR spectra dataset and solid-state NMR data of cowpea (Vigna unguiculata).

In this article the NMR data from chemical shifts, coupling constants, and structures of all the characterized compounds were provided, beyond a complementary PCA evaluation for the corresponding manuscript (E.G. Alves Filho, L.M.A. Silva, E.M. Teofilo, F.H. Larsen, E.S. de Brito, 2017) [3]. In addition, a complementary assessment from solid-state NMR data was provided. For further chemometric analysis, numerical matrices from the raw 1H NMR data were made available in Microsoft Excel workbook format (.xls).

Genotype evaluation of cowpea seeds (Vigna unguiculata) using 1H qNMR combined with exploratory tools and solid-state NMR.

The ultimate aim of this study was to apply a non-targeted chemometric analysis (principal component analysis and hierarchical clustering analysis using the heat map approach) of NMR data to investigate the variability of organic compounds in nine genotype cowpea seeds, without any complex pre-treatment. In general, both exploratory tools show that Tvu 233, CE-584, and Setentão genotypes presented higher amount mainly of raffinose and Tvu 382 presented the highest content of choline and least content of raffinose. The evaluation of the aromatic region showed the Setentão genotype with highest content of niacin/vitamin B3 whereas Tvu 382 with lowest amount. To investigate rigid and mobile components in the seeds cotyledon, 13C CP and SP/MAS solid-state NMR experiments were performed. The cotyledon of the cowpea comprised a rigid part consisting of starch as well as a soft portion made of starch, fatty acids, and protein. The variable contact time experiment suggests the presence of lipid-amylose complexes.

(1)H NMR spectroscopy and chemometrics evaluation of non-thermal processing of orange juice.

This study evaluated the effect of atmospheric cold plasma and ozone treatments on the key compounds (sugars, amino acids and short chain organic acids) in orange juice by NMR and chemometric analysis. The juice was directly and indirectly exposed to atmospheric cold plasma field at 70kV for different treatment time (15, 30, 45 and 60sec). For ozone processing different loads were evaluated. The Principal Component Analysis shown that the groups of compounds are affected differently depending on the processing. The ozone was the processing that more affected the aromatic compounds and atmospheric cold plasma processing affected more the aliphatic compounds. However, these variations did not result in significant changes in orange juice composition as a whole. Thus, NMR data and chemometrics were suitable to follow quality changes in orange juice processing by atmospheric cold plasma and ozone.

Characterization and quantitation of polyphenolic compounds in bark, kernel, leaves, and peel of mango (Mangifera indica L.).

The contents of secondary plant substances in solvent extracts of various byproducts (barks, kernels, peels, and old and young leaves) in a range of Brazilian mango cultivars were identified and quantitated. The results show that the profiles of secondary plant substances such as xanthone C-glycosides, gallotannins, and benzophenones in different byproducts vary greatly but are fairly consistent across cultivars. The free radical scavenging activity of the solvent extracts was evaluated using a high-performance liquid chromatography-based hypoxanthine/xanthine oxidase assay and revealed dose-dependent antioxidant capacity in all extracts. Four (mangiferin, penta- O-galloyl-glucoside gallic acid, and methyl gallate) of the major phenolic compounds detected were also evaluated in additional in vitro bioassay systems such as oxygen radical absorbance capacity, 2,2-diphenyl-1-picrylhydrazyl, and ferric reducing ability of plasma. Mangiferin in particular, detected at high concentrations in young leaves (Coite = 172 g/kg), in bark (Momika = 107 g/kg), and in old leaves (Itamaraka = 94 g/kg), shows an exceptionally strong antioxidant capacity.