1H-NMR Metabolomic Study of the Mushroom Pleurotus djamor for the Identification of Nematocidal Compounds.

Due to the increasing populations of anthelmintic-resistant gastrointestinal nematodes and as a consequence of the adverse effects of synthetic drugs, this study focuses on the search for secondary metabolites with nematocidal activity from the edible mushroom Pleurotus djamor using The proton nuclear magnetic resonance (1H-NMR) metabolomics. The highest activity was shown by the ethyl acetate fractions of mycelium (EC50 290.8 µg/mL) and basidiomes (EC50 282.7 µg/mL). Principal component analysis (PCA) and hierarchical data analysis (HCA) of the 1H-NMR metabolic profiles data showed that the ethanolic extracts, the ethyl acetate, butanol, and water fractions from mycelium have different metabolic profiles than those from basidiomes, while low polarity (hexane) fractions from both stages of fungal development show similar profiles. Orthogonal partial least squares discriminant analysis (OPLS-DA) allowed the identification of signals in the 1H-NMR metabolic profile associated with nematocidal activity. The signals yielded via OPLS-DA and bidimensional NMR analysis allowed the identification of uracil as a component in the ethyl acetate fraction from basidiomes, with an EC50 of 237.7 µg/mL. The results obtained showed that chemometric analyses of the 1H-NMR metabolic profiles represent a viable strategy for the identification of bioactive compounds from samples with complex chemical profiles.

HR-MAS NMR Metabolomics Profile of Vero Cells under the Influence of Virus Infection and nsP2 Inhibitor: A Chikungunya Case Study.

The arbovirus Chikungunya (CHIKV) is transmitted by Aedes mosquitoes in urban environments, and in humans, it triggers debilitating symptoms involving long-term complications, including arthritis and Guillain-Barré syndrome. The development of antiviral therapies is relevant, as no efficacious vaccine or drug has yet been approved for clinical application. As a detailed map of molecules underlying the viral infection can be obtained from the metabolome, we validated the metabolic signatures of Vero E6 cells prior to infection (CC), following CHIKV infection (CV) and also upon the inclusion of the nsP2 protease inhibitor wedelolactone (CWV), a coumestan which inhibits viral replication processes. The metabolome groups evidenced significant changes in the levels of lactate, myo-inositol, phosphocholine, glucose, betaine and a few specific amino acids. This study forms a preliminary basis for identifying metabolites through HR-MAS NMR (High Resolution Magic Angle Spinning Nuclear Magnetic Ressonance Spectroscopy) and proposing the affected metabolic pathways of cells following viral infection and upon incorporation of putative antiviral molecules.

Study of the Stability of Wine Samples for 1H-NMR Metabolomic Profile Analysis through Chemometrics Methods.

Wine is a temperature, light, and oxygen-sensitive product, so its physicochemical characteristics can be modified by variations in temperature and time when samples are either sampled, transported, and/or analyzed. These changes can alter its metabolomic fingerprinting, impacting further classification tasks and quality/quantitative analyses. For these reasons, the aim of this work is to compare and analyze the information obtained by different chemometric methods used in a complementary form (PCA, ASCA, and PARAFAC) to study 1H-NMR spectra variations of four red wine samples kept at different temperatures and time lapses. In conjunction, distinctive changes in the spectra are satisfactorily tracked with each chemometric method. The chemometric analyses reveal variations related to the wine sample, temperature, and time, as well as the interactions among these factors. Moreover, the magnitude and statistical significance of the effects are satisfactorily accounted for by ASCA, while the time-related effects variations are encountered by PARAFAC modeling. Acetaldehyde, formic acid, polyphenols, carbohydrates, lactic acid, ethyl lactate, methanol, choline, succinic acid, proline, acetoin, acetic acid, 1,3-propanediol, isopentanol, and some amino acids are identified as some of the metabolites which present the most important variations.

Integrated NMR and MS Analysis of the Plasma Metabolome Reveals Major Changes in One-Carbon, Lipid, and Amino Acid Metabolism in Severe and Fatal Cases of COVID-19.

Brazil has the second-highest COVID-19 death rate worldwide, and Rio de Janeiro is among the states with the highest rate in the country. Although vaccine coverage has been achieved, it is anticipated that COVID-19 will transition into an endemic disease. It is concerning that the molecular mechanisms underlying clinical evolution from mild to severe disease, as well as the mechanisms leading to long COVID-19, are not yet fully understood. NMR and MS-based metabolomics were used to identify metabolites associated with COVID-19 pathophysiology and disease outcome. Severe COVID-19 cases (n = 35) were enrolled in two reference centers in Rio de Janeiro within 72 h of ICU admission, alongside 12 non-infected control subjects. COVID-19 patients were grouped into survivors (n = 18) and non-survivors (n = 17). Choline-related metabolites, serine, glycine, and betaine, were reduced in severe COVID-19, indicating dysregulation in methyl donors. Non-survivors had higher levels of creatine/creatinine, 4-hydroxyproline, gluconic acid, and N-acetylserine, indicating liver and kidney dysfunction. Several changes were greater in women; thus, patients' sex should be considered in pandemic surveillance to achieve better disease stratification and improve outcomes. These metabolic alterations may be useful to monitor organ (dys) function and to understand the pathophysiology of acute and possibly post-acute COVID-19 syndromes.

Enzymatic and Synthetic Routes of Castor Oil Epoxidation.

Epoxidation of castor oil in synthetic and enzymatic routes was carried out in order to promote a system with less environmental impact. The epoxidation reactions of castor oil compounds upon addition of lipase enzyme with and without acrylic immobilization and with reaction times of 24 and 6 h, as well as the synthetic compounds upon addition of Amberlite resin and formic acid, were investigated using Fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance in hydrogen molecules (1H-NMR). The analysis indicated that the enzymatic reactions (6 h) and synthetic reactions provided a conversion from 50 to 96% and epoxidation from 25 to 48%, resulting from peak stretching and signal disintegration in the hydroxyl region due to the appearance of H2O in the interaction of peracid with catalyst. In systems without toluene, a dehydration event with a peak absorbance of 0.02 AU, indicating a possible vinyl group at 2355 cm-1 in enzymatic reactions without acrylic immobilization, was observed and resulted in a selectivity of 2%. In the absence of a solid catalyst, an unsaturation conversion of castor oil above 90% was achieved; however, this catalyst is necessary for the epoxidation to take place, whereas the lipase enzyme becomes able of epoxidizing and dehydrating the castor oil upon changing the time or reaction system. The conversation from 28 to 48% of solid catalysts (Amberlite and lipase enzyme) displays their importance to the instauration conversion of castor oil into oxirane rings.

Impact of genus (Geotrigona, Melipona, Scaptotrigona) in the targeted 1H-NMR organic profile, and authenticity test by interphase emulsion of honey processed in cerumen pots by stingless bees in Ecuador.

The biodiversity of Ecuadorian stingless bees is almost 200 species. Traditional pot-honey harvest in Ecuador is mostly done from nests of the three genera selected here Geotrigona Moure, 1943, Melipona Illiger, 1806, and Scaptotrigona Moure, 1942. The 20 pot-honey samples collected from cerumen pots and three ethnic honeys "abeja de tierra", "bermejo", and "cushillomishki" were analyzed for qualitative and quantitative targeted 1H-NMR honey profiling, and for the Honey Authenticity Test by Interphase Emulsion (HATIE). Extensive data of targeted organic compounds (41 parameters) were identified, quantified, and described. The three honey types were compared by ANOVA. Amino acids, ethanol, hydroxymethylfurfural, aliphatic organic acids, sugars, and markers of botanical origin. The number of phases observed with the HATIE were one in Scaptotrigona and three in Geotrigona and Melipona honeys. Acetic acid (19.60 ± 1.45 g/kg) and lactic acid (24.30 ± 1.65 g/kg) were particularly high in Geotrigona honey (in contrast to 1.3 g/kg acetic acid and 1.6 g/kg lactic acid in Melipona and Scaptotrigona), and with the lowest fructose + glucose (18.39 ± 1.68) g/100g honey compared to Melipona (52.87 ± 1.75) and Scaptotrigona (52.17 ± 0.60). Three local honeys were tested using PCA (Principal Component Analysis), two were assigned with a correct declared bee origin, but "bermejo" was not a Melipona and grouped with the Scaptotrigona cluster. However after HCA (Hierarchical Cluster Analysis) the three honeys were positioned in the Melipona-Scaptotrigona cluster. This research supports targeted 1H-NMR-based profiling of pot-honey metabolomics approach for multi-parameter visualization of organic compounds, as well as descriptive and pertained multivariate statistics (HCA and PCA) to discriminate the stingless bee genus in a set of Geotrigona, Melipona and Scaptotrigona honey types. The NMR characterization of Ecuadorian honey produced by stingless bees emphasizes the need for regulatory norms. A final note on stingless bee markers in pot-honey metabolites which should be screened for those that may extract phylogenetic signals from nutritional traits of honey. Scaptotrigona vitorum honey revealed biosurfactant activity in the HATIE, originating a fingerprint Honey Biosurfactant Test (HBT) for the genus in this set of pot-honeys.

Metabolomics by NMR Combined with Machine Learning to Predict Neoadjuvant Chemotherapy Response for Breast Cancer.

Neoadjuvant chemotherapy (NACT) is offered to patients with operable or inoperable breast cancer (BC) to downstage the disease. Clinical responses to NACT may vary depending on a few known clinical and biological features, but the diversity of responses to NACT is not fully understood. In this study, 80 women had their metabolite profiles of pre-treatment sera analyzed for potential NACT response biomarker candidates in combination with immunohistochemical parameters using Nuclear Magnetic Resonance (NMR). Sixty-four percent of the patients were resistant to chemotherapy. NMR, hormonal receptors (HR), human epidermal growth factor receptor 2 (HER2), and the nuclear protein Ki67 were combined through machine learning (ML) to predict the response to NACT. Metabolites such as leucine, formate, valine, and proline, along with hormone receptor status, were discriminants of response to NACT. The glyoxylate and dicarboxylate metabolism was found to be involved in the resistance to NACT. We obtained an accuracy in excess of 80% for the prediction of response to NACT combining metabolomic and tumor profile data. Our results suggest that NMR data can substantially enhance the prediction of response to NACT when used in combination with already known response prediction factors.

Inclusion Complex between Local Anesthetic/2-hydroxypropyl-ß-cyclodextrin in Stealth Liposome.

The drugs delivery system in the treatment of diseases has advantages such as reduced toxicity, increased availability of the drug, etc. Therefore, studies of the supramolecular interactions between local anesthetics (LAs) butamben (BTB) or ropivacaine (RVC) complexed with 2-hydroxypropyl-ß-cyclodextrin (HP-ßCD) and carried in Stealth liposomal (SL) are performed. 1H-NMR nuclear magnetic resonance (DOSY and STD) were used as the main tools. The displacements observed in the 1H-NMR presented the complexion between LAs and HP-ßCD. The diffusion coefficients of free BTB and RVC were 7.70 × 10-10 m2 s-1 and 4.07 × 10-10 m2 s-1, and in the complex with HP-ßCD were 1.90 × 10-10 m2 s-1 and 3.64 × 10-10 m2 s-1, respectively, which indicate a strong interaction between the BTB molecule and HP-ßCD (98.3% molar fraction and Ka = 72.279 L/mol). With STD-NMR, the encapsulation of the BTB/HP-ßCD and RVC/HP-ßCD in SL vesicles was proven. Beyond the saturation transfer to the LAs, there is the magnetization transfer to the hydrogens of HP-ßCD. BTB and RVC have already been studied in normal liposome systems; however, little is known of their behavior in SL.

High-Resolution Magic-Angle Spinning NMR Spectroscopy for Evaluation of Cell Shielding by Virucidal Composites Based on Biogenic Silver Nanoparticles, Flexible Cellulose Nanofibers and Graphene Oxide.

Antiviral and non-toxic effects of silver nanoparticles onto in vitro cells infected with coronavirus were evaluated in this study using High-Resolution Magic-Angle Spinning Nuclear Magnetic Resonance (HR-MAS NMR) spectroscopy. Silver nanoparticles were designed and synthesized using an orange flavonoid-hesperetin (HST)-for reduction of silver(I) and stabilization of as obtained nanoparticles. The bio-inspired process is a simple, clean, and sustainable way to synthesize biogenic silver nanoparticles (AgNP@HST) with diameters of ∼20 nm and low zeta potential (-40 mV), with great colloidal stability monitored for 2 years. The nanoparticles were used for the fabrication of two types of antiviral materials: colloids (AgNP@HST spray) and 3D flexible nanostructured composites. The composites, decorated with AgNP@HST (0.05 mmol L-1), were made using cellulose nanofibers (CNF) obtained from orange peel and graphene oxide (GO), being denominated CNF@GO@AgNP@HST. Both materials showed high virucidal activity against coronaviruses in cell infection in vitro models and successfully inhibited the viral activity in cells. HR-MAS 1H-NMR technique was used for determining nanomaterials' effects on living cells and their influences on metabolic pathways, as well as to study viral effects on cells. It was proven that none of the manufactured materials showed toxicity towards the intact cells used. Furthermore, viral infection was reverted when cells, infected with the coronavirus, were treated using the as-fabricated nanomaterials. These significant results open possibilities for antiviral application of 3D flexible nanostructured composite such as packaging papers and filters for facial masks, while the colloidal AgNP@HST spray can be used for disinfecting surfaces, as well as a nasal, mouth, and eye spray.

Ascidian (Chordata-Tunicata) Glycosaminoglycans: Extraction, Purification, Biochemical, and Spectroscopic Analysis.

Sulfate polysaccharides with unique structures of the chondroitin/dermatan and heparin/heparan families of sulfated glycosaminoglycans have been described in several species of ascidians (Chordata-Tunicata). These unique sulfated glycans have been isolated from the ascidians and characterized by biochemical and spectroscopic methods. The ascidian glycans can be extracted by different tissues or cells by proteolytic digestion followed by cetylpyridinium chloride/ethanol precipitation. The total glycans are then fractionated by ion-exchange chromatography on DEAE-cellulose and/or Mono Q (HR 5/5) columns. Alternatively, precipitation with different ethanol concentrations can be employed. An initial analysis of the purified ascidian glycans is carried out by agarose gel electrophoresis on diaminopropane/acetate buffer, before or after digestion with specific glycosaminoglycan lyases or deaminative cleavage with nitrous acid. The disaccharides formed by exhaustive degradation of the glycans are purified by gel-filtration chromatography on a Superdex Peptide column and analyzed by HPLC on a strong ion-exchange Sax Spherisorb column. 1H- or 13C-nuclear magnetic resonance spectroscopy in one or two dimensions is used to confirm the structure of the intact glycans.

1H -NMR Metabolomics Study of the Effect of Cisplatin and Casiopeina IIgly on MDA-MB-231 Breast Tumor Cells.

The knowledge of the metabolic processes of designed metallodrugs for cancer treatment is an area that has been not profoundly studied. Casiopeina IIgly (CasIIgly), which belongs to the Casiopeínas® family, is a copper (II) coordination compound that has shown good biological activity against several cancer cells, low toxicity in normal cells, and antineoplastic activity in in vivo murine and xenografted models. In this work we employed a triple-negative highly metastatic breast carcinoma line (MDA-MB-231), which is one of the cancer types with a great mortality index, for 1H-NMR metabolomic analysis using cisplatin and CasIIgly, in order to quantify the effect of metallodrugs in the metabolic profile of this cell tumor line as a consequence of treatment at different times. Our findings indicate that cisplatin mainly contributes to phospholipid biosynthesis while CasIIgly affects processes such as carbohydrates and nucleotides metabolism. Also, we observed that CasIIgly treatment has an important and fast effect over MDA-MB-231 cell metabolism, which makes it a good alternative for treatment in this type of cancer.

In vitro pharmacological attributes and metabolite's fingerprinting of Conocarpus lancifolius / Atributos farmacológicos in vitro y huellas dactilares de metabolitos de Conocarpus lancifolius

Search for safe antioxidants and novel nutraceuticals urged to evaluate the antioxidant, anti-acetylcholine esterase and anti-lipoxygenase activity of various leaf extracts of Conocarpus lancifolius. Extraction was optimized from freeze dried plant extracts quenched with liquid nitrogen using water, ethanol, methanol, hexane, ethyl acetate and chloroform. Maximum extract yield, total phenolic contents and total flavonoid contents were obtained in case of ethanolic extraction. The highest 2,2-diphenyl-1-picrylhydrazylradical scavenging in terms of IC50 value of 55.26 µg/mL was observed for ethanolic leaf extract. The acetylcholine esterase and lipoxygenase inhibitory activities (IC50) were also observed for ethanolic extract. These findings for ethanolic extract were statistically significant when compared with other extracts (ρ<0.05). The haemolytic % values indicated that all extracts were associated with very low or negligible toxicity. The epicatechin, isorhamnetin, rutin, scopoleptin, skimmianine, quercetin-3-O-α-rhamnoside, quercetin-3-O-ß-glucoside, cornoside, creatinine, choline, pyruvic acid, α-hydroxybutyric acid, phyllanthin and hypophyllanthin were identified as major functional metabolites in ethanolic leaf extract of C. lancifoliusby 1H-NMR. The identified metabolites were probably responsible for the pharmacological properties of C.lancifolius. The findings may be utilized as pharmacological leads for drug development and food fortification.

Se insta a la búsqueda de antioxidantes seguros y nuevos nutracéuticos para evaluar la actividad antioxidante, anti-acetilcolina esterasa y anti-lipoxigenasa de varios extractos de hojas de Conocarpus lancifolius. La extracción se optimizó a partir de extractos de plantas liofilizados enfriados con nitrógeno líquido usando agua, etanol, metanol, hexano, acetato de etilo y cloroformo. En el caso de extracción etanólica se obtuvo el rendimiento máximo de extracto, el contenido de fenoles totales y el contenido de flavonoides totales. La mayor eliminación de radicales 2,2-difenil-1-picrilhidrazilo en términos de valor de CI50 de 55,26 µg/mL se observó para el extracto de hoja etanólico. También se observaron las actividades inhibidoras de la acetilcolina esterasa y lipoxigenasa (CI50) para el extracto etanólico. Estos hallazgos para el extracto etanólico fueron estadísticamente significativos en comparación con otros extractos (ρ<0.05). Los valores del % hemolítico indicaron que todos los extractos estaban asociados con una toxicidad muy baja o insignificante. Se identificaron la epicatequina, isorhamnetina, rutina, escopoleptina, skimmianina, quercetina-3-O-α-ramnosido, quercetina-3-O-ß-glucósido, cornosido, creatinina, colina, ácido pirúvico, ácido α-hidroxibutírico, filantrina e hipofillantina. como metabolitos funcionales principales en el extracto etanólico de hojas de C. lancifoliuspor 1H-NMR. Los metabolitos identificados probablemente fueron responsables de las propiedades farmacológicas de C. lancifolius. Los hallazgos pueden utilizarse como pistas farmacológicas para el desarrollo de fármacos y la fortificación de alimentos.

Metabolomic Analysis of Combretum lanceolatum Plants Interaction with Diaporthe phaseolorum and Trichoderma spirale Endophytic Fungi through 1 H-NMR.

Endophytic fungi are an important class of microorganisms, able to interact with a host plant via a mutualistic mechanism without visible symptoms of the fungal colonization. The synergy between endophytic fungi and their host plant can promote morphological, physiological and biochemical changes through the expression of bioactive metabolites. This work aims to correlate metabolic changes in the Combretum lanceolatum plant metabolome with its endophytic fungi Diaporthe phaseolorum (Dp) and Trichoderma spirale (Ts), and to discover corresponding metabolite-biomarkers, with the principal focus being on its primary metabolism. The 1 H-NMR metabolomic analysis of qualitative and quantitative changes was performed through multivariate statistical analysis and the identification of primary metabolites was achieved on the Madison Metabolomics Consortium Database. The presence of Dp significantly impacted the plant's metabolic pathways, improving the biosynthesis of primary metabolites such as threonine, malic acid and N-acetyl-mannosamine, which are precursors of special metabolites involved in plant self-defence. This work represents a valuable contribution to advanced studies on the metabolic profiles of the interaction of plants with endophytes.

Nuclear Magnetic Resonance Metabolomics with Double Pulsed-Field-Gradient Echo and Automatized Solvent Suppression Spectroscopy for Multivariate Data Matrix Applied in Novel Wine and Juice Discriminant Analysis.

The quality of foods has led researchers to use various analytical methods to determine the amounts of principal food constituents; some of them are the NMR techniques with a multivariate statistical analysis (NMR-MSA). The present work introduces a set of NMR-MSA novelties. First, the use of a double pulsed-field-gradient echo (DPFGE) experiment with a refocusing band-selective uniform response pure-phase selective pulse for the selective excitation of a 5-10-ppm range of wine samples reveals novel broad 1H resonances. Second, an NMR-MSA foodomics approach to discriminate between wine samples produced from the same Cabernet Sauvignon variety fermented with different yeast strains proposed for large-scale alcohol reductions. Third a comparative study between a nonsupervised Principal Component Analysis (PCA), supervised standard partial (PLS-DA), and sparse (sPLS-DA) least squares discriminant analysis, as well as orthogonal projections to a latent structures discriminant analysis (OPLS-DA), for obtaining holistic fingerprints. The MSA discriminated between different Cabernet Sauvignon fermentation schemes and juice varieties (apple, apricot, and orange) or juice authentications (puree, nectar, concentrated, and commercial juice fruit drinks). The new pulse sequence DPFGE demonstrated an enhanced sensitivity in the aromatic zone of wine samples, allowing a better application of different unsupervised and supervised multivariate statistical analysis approaches.

Metabolite Biomarkers of Leishmania Antimony Resistance.

Leishmania parasites cause leishmaniasis, one of the most epidemiologically important neglected tropical diseases. Leishmania exhibits a high ability of developing drug resistance, and drug resistance is one of the main threats to public health, as it is associated with increased incidence, mortality, and healthcare costs. The antimonial drug is the main historically implemented drug for leishmaniasis. Nevertheless, even though antimony resistance has been widely documented, the mechanisms involved are not completely understood. In this study, we aimed to identify potential metabolite biomarkers of antimony resistance that could improve leishmaniasis treatment. Here, using L. tropica promastigotes as the biological model, we showed that the level of response to antimony can be potentially predicted using 1H-NMR-based metabolomic profiling. Antimony-resistant parasites exhibited differences in metabolite composition at the intracellular and extracellular levels, suggesting that a metabolic remodeling is required to combat the drug. Simple and time-saving exometabolomic analysis can be efficiently used for the differentiation of sensitive and resistant parasites. Our findings suggest that changes in metabolite composition are associated with an optimized response to the osmotic/oxidative stress and a rearrangement of carbon-energy metabolism. The activation of energy metabolism can be linked to the high energy requirement during the antioxidant stress response. We also found that metabolites such as proline and lactate change linearly with the level of resistance to antimony, showing a close relationship with the parasite's efficiency of drug resistance. A list of potential metabolite biomarkers is described and discussed.

In vitro impact of condensed tannins on the caecal metabolites of chickens / Impacto in vitro de taninos condensados nos metabólitos cecais de frangos de corte

Tannins are a diverse group of plant phenolic compounds. Condensed tannins (CTs) represent a major subgroup of tannins and were extracted from tilia (Tilia L.) flowers and black locust (Robinia pseudoacacia) leaves. These extracts were examined for their effects on the metabolic profile of chicken caeca. By using in vitro, a nuclear magnetic resonance (1H-NMR), which was combined with multivariate statistics, the current study was applied for the first time to investigate how three different CT compositions, procyanidins (PC) and/or prodelphinidins (PD) units influenced the metabolic end-products in caecal contents of chickens. In the presence of tannins, glutamate, leucine, lysine, pyroglutamate, phenylalanine, proline, and sarcosine were significantly decreased. CT extracts significantly influenced the fermentation, increasing the concentrations of some fatty acids such as acetate, butyrate, and propionate whereas. In contrast, lactate decreased between the treatments. This study identified the key structural features of CTs that contain either high molar proportions of PD or PC, which might be useful to improve the efficiency of feed utilization in chickens.(AU)

Taninos são um grupo diversificado de compostos fenólicos derivados de plantas. Os taninos condensados (TCs) representam o maior subgrupo de taninos extraídos das flores de tília (Tilia L) e de folhas negras (acácia-bastarda). Estes extratos foram examinados para a avaliação dos seus efeitos no perfil metabólico do ceco de frangos de corte. Com o emprego da ressonância magnética nuclear in vitro (1H-NMR) combinada com estatística multivariada, o presente trabalho foi aplicado pela primeira vez para investigar como três diferentes composições de TCs, unidades de procianidinas (PC) e/ou prodelfinidinas (PD) influenciariam os produtos metabólicos finais dos conteúdos cecais de frangos de corte. Na presença de taninos, houve um significativo decréscimo de glutamato, leucina, lisina, piroglutamato, fenilalanina, prolina e sarcosina. Os extratos de TCs influenciaram significativamente a fermentação, aumentando as concentrações de alguns ácidos graxos, tais como o acetato, butirato e propionato, enquanto em contraste, houve um decréscimo do lactato entre os tratamentos. Este trabalho identificou aspectos estruturais chave que os TCs contêm, tanto as altas proporções molares de PD como as de PC, as quais podem ser úteis para aumentar a utilização de alimentos em frangos de corte.(AU)

In vitro impact of condensed tannins on the caecal metabolites of chickens / Impacto in vitro de taninos condensados nos metabólitos cecais de frangos de corte

Tannins are a diverse group of plant phenolic compounds. Condensed tannins (CTs) represent a major subgroup of tannins and were extracted from tilia (Tilia L.) flowers and black locust (Robinia pseudoacacia) leaves. These extracts were examined for their effects on the metabolic profile of chicken caeca. By using in vitro, a nuclear magnetic resonance (1H-NMR), which was combined with multivariate statistics, the current study was applied for the first time to investigate how three different CT compositions, procyanidins (PC) and/or prodelphinidins (PD) units influenced the metabolic end-products in caecal contents of chickens. In the presence of tannins, glutamate, leucine, lysine, pyroglutamate, phenylalanine, proline, and sarcosine were significantly decreased. CT extracts significantly influenced the fermentation, increasing the concentrations of some fatty acids such as acetate, butyrate, and propionate whereas. In contrast, lactate decreased between the treatments. This study identified the key structural features of CTs that contain either high molar proportions of PD or PC, which might be useful to improve the efficiency of feed utilization in chickens.(AU)

Taninos são um grupo diversificado de compostos fenólicos derivados de plantas. Os taninos condensados (TCs) representam o maior subgrupo de taninos extraídos das flores de tília (Tilia L) e de folhas negras (acácia-bastarda). Estes extratos foram examinados para a avaliação dos seus efeitos no perfil metabólico do ceco de frangos de corte. Com o emprego da ressonância magnética nuclear in vitro (1H-NMR) combinada com estatística multivariada, o presente trabalho foi aplicado pela primeira vez para investigar como três diferentes composições de TCs, unidades de procianidinas (PC) e/ou prodelfinidinas (PD) influenciariam os produtos metabólicos finais dos conteúdos cecais de frangos de corte. Na presença de taninos, houve um significativo decréscimo de glutamato, leucina, lisina, piroglutamato, fenilalanina, prolina e sarcosina. Os extratos de TCs influenciaram significativamente a fermentação, aumentando as concentrações de alguns ácidos graxos, tais como o acetato, butirato e propionato, enquanto em contraste, houve um decréscimo do lactato entre os tratamentos. Este trabalho identificou aspectos estruturais chave que os TCs contêm, tanto as altas proporções molares de PD como as de PC, as quais podem ser úteis para aumentar a utilização de alimentos em frangos de corte.(AU)

1 H-NMR and GC for detection of adulteration in commercial essential oils of Cymbopogon ssp.

INTRODUCTION: Essential oils of Cymbopogon nardus and C. winterianus have fungicidal, bactericidal, and insect repellent activities. In addition, they are components of fragrances, cosmetics, and household products. The growing demand for essential oils has intensified adulteration practices of such products. OBJECTIVES: To evaluate the authenticity and quality of citronella commercial essential oils based on chemical composition [by gas chromatography mass spectrometry (GC-MS)] and the contents of its major constituents [by 1 H-NMR, and gas chromatography with a flame ionisation detector using internal standardisation (GC-IS)]. MATERIALS AND METHODS: The chemical composition of essential oil was determined by GC-MS. Major components were quantified by 1 H-NMR and the results compared to those obtained by GC-IS. RESULTS: The adulteration of oils was verified by GC and 1 H-NMR. In the pure oils, the results obtained by 1 H-NMR were similar to those obtained by GC-IS for most of the oils. However, in adulterated oils, signal overlap prevented the quantification of citronellol and geraniol by NMR. Importantly, due to dilution with dipropylene glycol it was not possible to quantify citronellal using 1 H-NMR. However, for both pure and adulterated oils, GC-IS method proved successful in quantifying notable constituents. CONCLUSION: All the methods used proved efficient in detecting adulteration. However, whilst GC-IS provided quantification of constituents of interest, both in pure and adulterated oils, their quantification by NMR was only possible in non-adulterated samples. None of the oils evaluated presented a composition within the threshold established by British Pharmacopoeia quality standards.

The Influence of Drying Temperatures on the Metabolic Profiles and Antioxidant Activity of Manilkara zapota Leaves.

In the present study, the leaves of Manilkara zapota (L.) P. Royen (Sapotaceae), an evergreen tree recognized for its medicinal properties in Southern Mexico, were used as a model to study the effect of different drying temperatures on its metabolic profile and therefore, its antioxidant potential. For this purpose, a methanol extraction of leaves dried at room temperature (25 °C) or by heat convection (50, 75 and 100 °C) were compared in terms of drying efficiency, yield of extraction, total phenol content, 1H-NMR metabolic profile, and DPPH antioxidant activity. The drying curves enabled the fact to be uncovered that drying efficiency improves with increase of temperature, as does the level of total phenols and antioxidant activity. A metabolomics approach using principal component analysis (PCA) and orthogonal projections to latent structures discriminant analysis (OPLS-DA) of the corresponding 1H-NMR profiles allowed the impact of the drying temperature on their metabolic profile to be documented and also, caffeic acid and epicatechin as main secondary metabolites contributing to the antioxidant activity of M. zapota to be identified.

Improvement of bioactive metabolite production in microbial cultures-A systems approach by OSMAC and deconvolution-based 1 HNMR quantification.

Traditionally, the screening of metabolites in microbial matrices is performed by monocultures. Nonetheless, the absence of biotic and abiotic interactions generally observed in nature still limit the chemical diversity and leads to "poorer" chemical profiles. Nowadays, several methods have been developed to determine the conditions under which cryptic genes are activated, in an attempt to induce these silenced biosynthetic pathways. Among those, the one strain, many compounds (OSMAC) strategy has been applied to enhance metabolic production by a systematic variation of growth parameters. The complexity of the chemical profiles from OSMAC experiments has required increasingly robust and accurate techniques. In this sense, deconvolution-based 1 HNMR quantification have emerged as a promising methodology to decrease complexity and provide a comprehensive perspective for metabolomics studies. Our present work shows an integrated strategy for the increased production and rapid quantification of compounds from microbial sources. Specifically, an OSMAC design of experiments (DoE) was used to optimize the microbial production of bioactive fusaric acid, cytochalasin D and 3-nitropropionic acid, and Global Spectral Deconvolution (GSD)-based 1 HNMR quantification was carried out for their measurement. The results showed that OSMAC increased the production of the metabolites by up to 33% and that GSD was able to extract accurate NMR integrals even in heavily coalescence spectral regions. Moreover, GSD-1 HNMR quantification was reproducible for all species and exhibited validated results that were more selective and accurate than comparative methods. Overall, this strategy up-regulated important metabolites using a reduced number of experiments and provided fast analyte monitor directly in raw extracts.