Technical feasibility of an ecofriendly sequential process (ultrasound assisted extraction and reverse osmosis, or UAE and RO) was evaluated in order to obtain a functional Camu-camu (Myrciaria dubia) product with high vitamin C content. Water was used in the assisted extraction by probe ultrasound (UAE) in an experimental design to evaluate different times, amplitudes and temperatures. The best region for total phenolic (TP) and vitamin C (VC) extraction was 5 min, 60 °C and 30% amplitude. Following extraction, the sample was concentrated by reverse osmosis (R25a, 500 Da, polyamide, and 5 bar area 3 ft2), obtaining a relatively low fouling of 19%. At the end of the sequential process (by HPLC-DAD/UV vis), was obtained a concentrated camu-camu (CC) with high Vitamin C (52.01 ± 0.889 mg/g) and cyanidin-3-glucoside, being respectively 7.0 and 4.5 times higher; also the concentration of phenolic compounds was increased by 3.2 times (25.798 mg GAE/g), and anthocyanins in 6.5 times (66.169 mg of cyanidin-3-glucoside/100 g) as well as high antioxidant activity by all three methods evaluated (increased 3.0, 4.6 and 2.38 times for ABTS, DPPH, FRAP, respectively) by comparing the CC with the initial extract (CS). Twenty compounds were identified by UHPLC-QTOF-MS/MS, highlighting quercetin, gallic acid, p- coumaric, ellagic acid and cyanidin-3-glucoside, and at the first time alnusiin was detected in camu-camu. Therefore, the combination of ultrasound assisted extraction and reverse osmosis can be a promising profitable alternative in order to apply bioactive compounds in food, nutraceuticals and cosmetic matrices, bringing their benefits to consumers.

Phakopsora pachyrhizi is a biotrophic fungus, causer of the disease Asian Soybean Rust, a severe crop disease of soybean and one that demands greater investment from producers. Thus, research efforts to control this disease are still needed. We investigated the expression of metabolites in soybean plants presenting a resistant genotype inoculated with P. pachyrhizi through the untargeted metabolomics approach. The analysis was performed in control and inoculated plants with P. pachyrhizi using UHPLC-MS/MS. Principal component analysis (PCA) and the partial least squares discriminant analysis (PLS-DA), was applied to the data analysis. PCA and PLS-DA resulted in a clear separation and classification of groups between control and inoculated plants. The metabolites were putative classified and identified using the Global Natural Products Social Molecular Networking platform in flavonoids, isoflavonoids, lipids, fatty acyls, terpenes, and carboxylic acids. Flavonoids and isoflavonoids were up-regulation, while terpenes were down-regulated in response to the soybean–P. pachyrhizi interaction. Our data provide insights into the potential role of some metabolites as flavonoids and isoflavonoids in the plant resistance to ASR. This information could result in the development of resistant genotypes of soybean to P. pachyrhizi, and effective and specific products against the pathogen.

Asian Soybean Rust (ASR), caused by the biotrophic fungus Phakopsora pachyrhizi, is a devastating disease with an estimated crop yield loss of up to 90%. Yet, there is a nerf of information on the metabolic response of soybean plants to the pathogen Untargeted metabolomics and Global Natural Products Social Molecular Networking platform approach was used to explore soybean metabolome modulation to P. pachyrhizi infection. Soybean plants susceptible to ASR was inoculated with P. pachyrhizi spore suspension and non-inoculated plants were used as controls. Leaves from both groups were collected 14 days post-inoculation and extracted using different extractor solvent mixtures. The extracts were analyzed on an ultra-high performance liquid chromatography system coupled to high-definition electrospray ionization-mass spectrometry. There was a significant production of defense secondary metabolites (phenylpropanoids, terpenoids and flavonoids) when P. pachyrhizi infected soybean plants, such as putatively identified liquiritigenin, coumestrol, formononetin, pisatin, medicarpin, biochanin A, glyoceollidin I, glyoceollidin II, glyoceollin I, glyoceolidin II, glyoceolidin III, glyoceolidin IV, glyoceolidin VI. Primary metabolites (amino acids, peptides and lipids) also were putatively identified. This is the first report using untargeted metabolomics and GNPS-Molecular Networking approach to explore ASR in soybean plants. Our data provide insights into the potential role of some metabolites in the plant resistance to ASR, which could result in the development of resistant genotypes of soybean to P. pachyrhizi, and effective and specific products against the pathogen.