Penicillium meliponae, a recently described and rare species, was isolated as an endophytic fungus from the Amazonian plant Duguetia sthelechantha, and has been proven to be a pigment producer. Considering the high productivity of this species and the lack of data on its chemical composition, the present study aimed to characterize the chemical profile of P. meliponae and evaluate the influence of agitation and the use of different culture media. For this purpose, liquid chromatography coupled with mass spectrometry (LC-MS/MS) and molecular networking were used, allowing the identification of 17 azaphilone molecules with sclerotiorin-like skeletons, becoming the first chemical report of this species. In addition, the different production patterns in the tested culture media were indicative that this species is sensitive to changes in the composition of the carbon source and to the presence of agitation. Furthermore, this work contributes to the fragmentation mechanisms of the different possible structural arrangements for azaphilones of the sclerotiorin type and serves as a repository of information on the gas-phase behavior of this type of metabolite in mass spectrometry experiments and will assist future studies aimed at the discovery of azaphilones.

The aim of the current study was the valorization of hesperidin, the dominant flavonoid in citrus processing waste, by microwave-assisted synthesis of hesperidin-Mg complex, improving its antioxidant activity and insecticidal potential. Here we show, for the first time, that microwave-assisted synthesis of [Mg(hesp)2(phen)]OAc (1) (hesp: hesperidin, phen: phenanthroline) improve the reaction rate and yield. The nuclear magnetic resonance (NMR) experiments proved to be powerful tools for the identification of three isomers in metal complexes. Moreover, we explore the insecticidal potential of 1 and [Mg(phen)2(Isov)]OAc (2) (Isov : isovanillic acid) complexes against three insects. Complex 1 killed 80% of adults whitefly at 0.14 μmol L-1, and 2 76% at 0.36 μmol L-1. There was a total mortality of Spodoptera frugiperda with 2 at 0.39 μmol L-1, and 83% with 1 at 0.14 μmol L-1. Both have similar activity, and represent a novel group of insecticide for Bemisia tabaci and S. frugiperda; nevertheless, the benefits of both as Myzus persicae repellent require further evaluation.

Sustainable natural product-based microstructured systems and biopolymers are strong candidates for use in crop protection. Lignin and gelatin microcapsules loaded with orange essential oil were developed with spray-drying in order to enhance its potential. We evaluated the microparticle controlled release mechanisms, biological effects, structural and thermal properties. A quantitative method using headspace-gas chromatography was developed and evaluated for the controlled release of the essential oils. Controlled release studies showed a linear relationship between the biopolymer and essential oil concentrations when retaining volatile compounds. Thermal analyses demonstrated increases in essential oil stability when microencapsulated, especially for lignin as a biopolymer. These results showed that biopolymer type was the main factor influencing quality analysis parameters. Finally, microcapsules loaded with orange essential oil were applied in a bioassay, and showed gains in toxicity against Spodoptera frugiperda compared to non-encapsulated oil.

Citrus biomass is a source of biobased products and presents extensive chemical diversity. Among the structural macromolecules, lignins have been used in resins, bioplastics, and nanoencapsulation. Brazil has a large source of lignins, which are orange trees removed from groves that need to be renewed. However, the knowledge about these lignins is limited and little explored. Thus, this work aimed to evaluate the lignins from Citrus sinensis and to provide information on the interesting use of orange trees as a source of macromolecules. The lignins were isolated from the trunk, central and secondary roots from the orange tree by Technical Association of the Pulp and Paper Industry (TAPPI)-methods. The lignin contents for the extractive-free samples were 19.8 ± 0.19, 21.9 ± 0.36, and 19.5 ± 0.41% (m/m) in the trunk, central and secondary roots, respectively. Infrared (IR) and 13C nuclear magnetic resonance (NMR) analyses confirmed typical bands and chemical shifts to syringyl and guaiacyl units. These results are consistent with the features of lignins previously reported for orange tree branches and hardwood roots. The greatest amount of lignin in the orange tree was found in the central root. The lignins extractive-free by TAPPI-T264 cm-97 presented low solubility in organic solvents, due to their compaction and folding, which was confirmed by 13C NMR in the solid-state, differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA).

Molecular phylogenetic studies separated and united a group of genera that constituted the Spathelia-Ptaeroxylon clade, in which Dictyoloma and Sohnreyia have been included. Our taxonomic interest in the Dictyoloma vandellianum and Sohnreyia excelsa stimulated an investigation of both species searching for limonoids. Leaves from D. vandellianum afforded the new limonoid 1,2-dihydro-1α-hydroxy-8,30-epoxy-cneorin R, and heartwood yielded the new rearranged limonoid dictyolomin. Leaves from S. excelsa afforded the new protolimonoid 3β-angeloyloxy-7α,24,25-trihydroxy-21,23-oxide-14,18-cycloapotirucall-21-methoxycetal and the new cycloheptanyl ring C limonoid with carbonate substituent and named as sohnreyolide. The new limonoids from Sohnreyia and Dictyoloma show similarities with those from Rutaceae and Meliaceae, providing support for moving Spathelia-Ptaeroxylon clade near to these associated large families.

Currently, most food waste is used as animal feed and this process does not take advantage of the available chemical composition. A Brazilian example is the citrus fruit processing wastes (CFPW), which have begun to draw attention due to their biological importance. In order to access the main compounds of this matrix, an efficient and environmentally friendly procedure was tested. From this extract, flavonoids as naringenin, hesperitin, chrysoeriol, sinensetin, 3,5,6,7,3',4'-hexamethoxyflavone, nobiletin, 5-methoxysalvigenin, 3,5,6,7,8,3',4'-heptamethoxyflavone, 3,5,6,7,4'-pentamethoxyflavone and isosakuranetin were identified by high performance liquid chromatography (HPLC-UV), liquid chromatography-mass spectrometry (LC-MS) and liquid chromatography-nuclear magnetic resonance (LC-NMR). This study describes the potential use of citrus waste as a source of biologically active compounds. The extraction method proposed for this work was not expensive and the flavonoids were obtained in large amounts, thus, this extraction method is being developed using pilot plant scale-up techniques and will soon be available to the industry at a low cost.

The continuous search for biologically active compound candidates pushes the pursuit of new substances produced by diverse organisms. Endophytic fungi are known as a promising source of metabolites with several biological activities. Based on the rich Amazonian biodiversity and the chemical potential of microbial sources, three Talaromyces strains, all major endophytes from their respective host plants, were studied aiming at the isolation of biologically active secondary metabolites. Through classical chromatographic approaches, 13 compounds were isolated from the antimicrobial extracts of the studied strains. From these, polyketides, steroids, anhydrides, and phenolic compounds were identified. Among them, two previously undescribed compounds were elucidated based on spectroscopic and spectrometric methods, a homodimer chromanone and a maleic anhydride methyl ester. The antimicrobial activity against a panel of pathogenic microorganisms from extracts, fractions, and isolated compounds was evaluated.

Microcapsules of sugarcane bagasse lignin loaded with organic extracts of neem (Azadirachta indica) were prepared and evaluated as potential bioinsecticides. Lignins were extracted and modified by oxidation and acetylation reactions providing different biopolymers. Afterwards, they were characterized through several analytical techniques. The formulations were initially prepared as colloidal suspension of lignin nanoparticles, which were then spray-dried. The products were submitted to quality control protocols using high performance liquid chromatography (HPLC), thermal and photochemical degradation studies, followed by biological assays against Spodoptera frugiperda and Diatraea saccharalis. The formulations showed increases in both thermal and photo stability of approximately 40% compared with control samples. All the formulated microparticles were efficient against the insects evaluated and required a shorter time to achieve 100% mortality than the controls. The use of sugarcane bagasse lignins to prepare microparticles improved the resistance of neem extracts against abiotic factors, retaining the biological activity of this biopesticide.

A methanol-soluble fraction of the dichloromethane extract from the culture broth of A. alternata AT4303 strain afforded the following five known mycotoxins: alternariol, alternariol monomethyl ether, altenusin, altenuene and altertoxin I. A hexane-soluble fraction of the dichloromethane extract yielded the steroids ergosterol and ergosterol peroxide, and alternariol monomethyl ether. The concentrated ethyl acetate extract afforded the nucleoside uridine, uracil and inosine. The micro-extracts obtained from this strain in solid media were analyzed using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS), and the following five known toxins were identified: ACTG-C, D, E and F and AK-toxin II. The major toxins produced by the tangerine pathotype ACT-toxins appear to be absent in strain AT4303. However, the low concentration of ACT-toxins appears to be responsible for their lack of detection in this study. Some compounds isolated from citrus plants were tested for in vitro activity against this strain. The results indicated that 17.3 µM apigenin-7-O-rutinoside inhibited by 80% conidial germination and appressorium development.

Plant flavonoids have been increasingly studied for its importance in plant metabolism, inter-species interactions and human health-promoting effects. Each botanical species has a distinct flavonoid profile, and as such, this work aimed to develop and apply a high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) method to quantify specific flavonoids in different plants and parts. Six flavonoids (isoquercitrin, quercetin, naringin, naringenin, hesperidin and rutin), two isoflavonoids (daidzin and daidzein) and one coumarin (umbelliferone) were targeted. Calibration curves showed good correlations (R2 > 0.994) and limits of quantification (≤ 500 ng mL-1). The method was applied for detection of analytes in common bean seeds (Phaseolus vulgaris), soybean leaves (Glycine max), and sour orange (Citrus aurantium), sweet orange (Citrus sinensis), and Tahiti lime (Citrus latifolia) flavedo after ultrasound assisted acidic hydroalcoholic extraction. The method was tuned for quick and efficient extraction. Principal component analysis was helpful for the discrimination of matrices based on their flavonoid profiles.