Eduardo Jorge Pilau

Phytochemical Analysis of Pfaffia glomerata Inflorescences by LC-ESI-MS/MS

Pfaffia glomerata contains high levels of β-ecdysone, which has shown a range of beneficial pharmacological effects. The present study demonstrated that inflorescences of P. glomerata contain other important bioactive compounds in addition to β-ecdysone. The identification of compounds from inflorescences using liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) was performed for the first time. The eight compounds identified were β-ecdysone, flavonoid glycosides such as quercetin-3-O-glucoside, kaempferol-3-O-glucoside and kaempferol-3-O-(6-p-coumaroyl)-glucoside, oleanane-type triterpenoid saponins such as ginsenoside Ro and chikusetsusaponin IV, in addition to oleanonic acid and gluconic acid. This study provided information on the phytochemicals contained in P. glomerata inflorescences revealing the potential application of this plant part as raw material for the phytotherapeutic and cosmetic industries.

Chemical characterization and prebiotic activity of fructo-oligosaccharides from Stevia rebaudiana (Bertoni) roots and in vitro adventitious root cultures.

Stevia rebaudiana (Bertoni) is widely studied because of its foliar steviol glycosides. Fructan-type polysaccharides were recently isolated from its roots. Fructans are reserve carbohydrates that have important positive health effects and technological applications in the food industry. The objective of the present study was to isolate and characterize fructo-oligosaccharides (FOSs) from S. rebaudiana roots and in vitro adventitious root cultures and evaluate the potential prebiotic effect of these molecules. The in vitro adventitious root cultures were obtained using a roller bottle system. Chemical analyses (gas chromatography-mass spectrometry, (1)H nuclear magnetic resonance, and off-line electrospray ionization-mass spectrometry) revealed similar chemical properties of FOSs that were obtained from the different sources. The potential prebiotic effects of FOSs that were isolated from S. rebaudiana roots enhanced the growth of both bifidobacteria and lactobacilli, with strains specificity in their fermentation ability.

Development of an analytical method for identification of Aspergillus flavus based on chemical markers using HPLC-MS

Aspergillus flavus is a filamentous fungus found in nature and characterized by the production of bright and colourful colonies. It grows on different substrates, producing secondary metabolites and, if present in foodstuffs, can be a source of health problems for humans and animals, as well as causing economic losses. Traditional methods for fungal identification are based on morphological characteristics, requiring specialists and being very time-consuming. The development of analytical alternatives might have advantages such as greater efficiency, more reproducibility and be less time-consuming. Thus, a qualitative analytical method to detect Aspergillus flavus in food samples, based on the identification of fungal chemical markers by HPLC-MS, was developed. The method comprises methanol extraction followed by HPLC-MS analysis, and was able to identify 14 fungus secondary metabolites, namely aflatoxin B1, aflatoxin G1, aspergillic acid, aspyrone, betaine, chrysogine, deacetyl parasiticolide A, flufuran, gregatin B, hydroxysydonic acid, nicotinic acid, phomaligin A, spinulosin and terrein.

Identification and ultra‐high‐performance liquid chromatography coupled with high‐resolution mass spectrometry characterization of biosurfactants, including a new surfactin, isolated from oil‐contaminated environments

Biosurfactant‐producing bacteria were isolated from samples collected in areas contaminated with crude oil. The isolates were screened for biosurfactant production using qualitative drop‐collapse test, oil‐spreading and emulsification assays, and measurement of their tensoactive properties. Five isolates tested positive for in the screening experiments and displayed decrease in the surface tension below 30 mN m−1. The biosurfactants produced by these isolates were further investigated and their molecular identification revealed that they are bacteria related to the Bacillus genus. Additionally, the biosurfactants produced were chemically characterized via UHPLC‐HRMS experiments, indicating the production of surfactin homologues, including a new class of these molecules.

Metabolomics and Agriculture: What Can Be Done?

The importance of Brazil as a producer and exporter of food and feed will continuously increase. Despite the recent economic and political problems in Brazil, the scientific field is expanding. ABSTRACT The importance of Brazil as a producer and exporter of food and feed will continuously increase. Despite the recent economic and political problems in Brazil, the scientific field is expanding. Cutting-edge technology has only recently become available in the country, and we can now also join efforts with the global community to tackle global challenges. Using metabolomics based on mass spectrometry approaches to understand system-wide metabolism and metabolic pathways can have a significant impact in the society. With the recent development of a platform for organization and sharing of tandem mass spectrometry data, the global community can now work with complex biological samples. In this Perspective, we aim to describe how challenges and problems in global and local agriculture can be addressed using metabolomics based on mass spectrometry strategies.

Pretreatment with ethanol as an alternative to improve steviol glycosides extraction and purification from a new variety of stevia

Leaves of a new variety of Stevia rebaudiana with a high content of rebaudioside A were pretreated with ethanol. The ethanolic extract showed high antioxidant potential and 39 compounds were identified, by UPLC/HRMS, among them one not yet mentioned in the literature for stevia leaves. From the in natura leaves and pretreated leaves, the conditions of aqueous extraction of steviol glycosides were investigated using response surface methodology. The aqueous extracts obtained were purified by ion exchange chromatography techniques and membrane separation methods. The recuperation of steviol glycosides was 4.02g for pretreated leaves and 2.20g for in natura leaves. The level of purity was, respectively, 87% and 84.8%. The results obtained demonstrate that pretreatment increases the yield and purity level of stevia sweeteners by the use of environmentally friendly methodologies and the final product presented acceptable sensory characteristics.

Biotransformation of (+)-carvone and (−)-carvone using human skin fungi: A green method of obtaining fragrances and flavours

Abstract The synthesis of optically pure compounds is increasingly in demand among the pharmaceutical, fine chemical and agro-food industries, while the importance of chirality in the activity and biological properties of many compounds has previously been established. The aim of the present study was therefore to evaluate the biotransformation capacities of (+)-carvone and (−)-carvone using the fungi Scolecobasidium sp, three lines of Cladosporium sp, Phoma sp, Aureobasidium sp and Epicoccum sp, all obtained from human skin. The seven fungi evaluated were capable of hydrogenating the activated alkene, followed by the reduction of ketone to chiral alcohol, with conversions between 9.5 and 100%, and with diastereomer excess (d.e.) of over 89% of dihydrocarveol when (+)-carvone was used as a substrate. These results demonstrate that the filamentous fungi of human skin are potential biocatalytic tools for obtaining chiral alcohols.

Fortification of the whey protein isolate antioxidant and antidiabetic activity with fraction rich in phenolic compounds obtained from Stevia rebaudiana (Bert.). Bertoni leaves

AbstractA stevia fraction (ASF) free of steviol glycosides was extracted from Stevia rebaudiana leaves (Stevia UEM-13). ASF essentially constitutes phenolic compounds (52.42%), which were identified by liquid chromatography tandem mass spectrometry (LC–MS/MS) as caffeic acid, quercetin-3-o-glycoside, cyanidin-3-glucoside, kaempferol, quercetin, apigenin, rozmarinic acid, chlorogenic acid and dicaffeoylquinic acid. ASF was used as a multi-functional source of phenolic compounds to fortify the whey protein isolate (WPI) obtained by membrane separation. WPI fortified with 0.2% ASF showed an 80% increase in its antioxidant activity and more pronounced antidiabetic effects than the unfortified WPI, mainly in the glycemic control of diabetic animals induced by streptozotocin. The in vitro and in vivo antioxidant effects of ASF may enhance the effects of WPI. Indeed, this pioneering study revealed that ASF can be used to enrich the antioxidant and antidiabetic properties of WPI.

CHEMICAL COMPOSITIONS AND ANTIOXIDANT AND ANTIMICROBIAL ACTIVITIES OF PROPOLIS PRODUCED BY Frieseomelitta longipes AND Apis mellifera BEES

Edineide Cristina A. de Souzaa, Etyene Janyne G. da Silvaa, Hayron Kalil C. Cordeirob, Nauara M. Lage Filhob, Felipe M. A. da Silvac, Diany Lucy S. dos Reisd, Carla Portod, Eduardo J. Pilaud, Luiz Antonio M. A. da Costaa, Afonso D. L. de Souzac, Cristiano Menezese and Adriana Flacha,*¤ Departamento de Química, Universidade Federal de Roraima, 69310-000, Boa Vista – RR, Brasil Universidade Federal Rural da Amazônia, 66075-110 Belém – PA, Brasil Departamento de Química, Universidade Federal da Amazônia, 69077-000, Manaus – AM, Brasil Departamento de Química, Universidade Estadual de Maringá, 87020-900, Maringá – PR, Brasil Embrapa Amazônia Oriental, 66095-903 Belém – PA, Brasil

Addressing Global Ruminant Agricultural Challenges Through Understanding the Rumen Microbiome: Past, Present, and Future

The rumen is a complex ecosystem composed of anaerobic bacteria, protozoa, fungi, methanogenic archaea and phages. These microbes interact closely to breakdown plant material that cannot be digested by humans, whilst providing metabolic energy to the host and, in the case of archaea, producing methane. Consequently, ruminants produce meat and milk, which are rich in high-quality protein, vitamins and minerals, and therefore contribute to food security. As the world population is predicted to reach approximately 9.7 billion by 2050, an increase in ruminant production to satisfy global protein demand is necessary, despite limited land availability, and whilst ensuring environmental impact is minimized. Although challenging, these goals can be met, but depend on our understanding of the rumen microbiome. Attempts to manipulate the rumen microbiome to benefit global agricultural challenges have been ongoing for decades with limited success, mostly due to the lack of a detailed understanding of this microbiome and our limited ability to culture most of these microbes outside the rumen. The potential to manipulate the rumen microbiome and meet global livestock challenges through animal breeding and introduction of dietary interventions during early life have recently emerged as promising new technologies. Our inability to phenotype ruminants in a high-throughput manner has also hampered progress, although the recent increase in “omic” data may allow further development of mathematical models and rumen microbial gene biomarkers as proxies. Advances in computational tools, high-throughput sequencing technologies and cultivation-independent “omics” approaches continue to revolutionize our understanding of the rumen microbiome. This will ultimately provide the knowledge framework needed to solve current and future ruminant livestock challenges.