Investigating specialized plant metabolites, traditionally referred to as ‘secondary metabolites’ present in leaf extracts of cashew trees (Anacardium occidentale) resistant and susceptible to anthracnose disease was carried out using metabolomics combined with chemometric tools. We used clones of dwarf-cashew with the following variations of characteristics: resistant and healthy (CCP 76, BRS 226, BRS 189), susceptible and healthy (BRS 265), and another clone also susceptible but affected by the disease (BRS 265). The UPLC-QTOF-MSE (ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry) analysis of the leaves of Anacardium clones allowed us to annotate a total of 39 metabolites. The multiple orthogonal partial discriminant analysis of the least-squares (OPLS-DA) allowed twelve metabolites to be potential biomarkers of differentiation among the clones studied. Namely, the triene-(17:3)-anacardic acid found on CCP 76 and BRS 189 clones, respectively, is the main candidate for biomarker of resistance. While catechin, B-type procyanidin isomers, and procyanidin dimer monogallate identified in BRS 265 are significant potential biomarkers of susceptibility.

Melon is a pollinator-dependent crop that relies mainly on bees to set fruits. However, man-selected varieties vary in their attractiveness to flower visitors, and some flower genders and melon types may be less visited than others, harming pollination. We investigated the nectar composition of male, female and hermaphrodite flowers of 270 individuals of five commercial melons Cucumis melo (Cantaloupe, Charentais, Galia, Piel de Sapo, and Yellow), and its role in flower visit by Apis mellifera foragers. We found that melon nectar is composed mainly of sugars and amino acids such as tyrosine, phenylalanine, tryptophan, and flavonoids kaempferol-3-O-neohesperidoside, luteolin hexoside, and kaempferol rhamnoside. But the amount of these chemical compounds varies among the flower genders. We also developed an accurate regression model to predict the number of bee visits to melon flowers based on the nectar composition. Our results indicate that nectar composition plays little role in bee discrimination among flowers of different melon types but is essential to the honeybee choice between flower gender. The amounts of phenylalanine (49.40%) and tryptophan (12.05%) in the nectar are related to bee preference for hermaphrodite flowers. More visits to hermaphrodite flowers contribute to setting and developing well-formed fruits, increasing productivity.

Crotalaria ochroleuca (Fabaceae) is rich in bioactive compounds used for nematode control. Transformed root culture allows the production of a large amount of hairy root biomass, conditioned by auxin biosynthesis-related genes integrated via the process of Agrobacterium rhizogenes infection, which is known to cause a hairy root phenotype. We transformed A. rhizogenes to increase the biomass of hairy roots and optimized the process of production of bioactive compounds. The transformed nature of hairy roots was confirmed by polymerase chain reaction, which confers kanamycin resistance. The transformed roots were cultured in full-strength in liquid media to realize the growth of hairy roots and the production of bioactive compounds. The transformed roots were grown in a culture medium supplemented with elicitors to produce total phenols, and flavonoids. Chromatographic analysis of transformed roots revealed the presence of flavonoids apigenin-6,8-C-diglucoside and luteolin-6-C-glucoside. The results were obtained by conducting physiological and biochemical studies with the Flavonoids and studying the pathways that led to the production of large amounts of bioactive compounds from the hairy roots of C. ochroleuca. It was observed that the extraction of the compounds significantly affected nematodes and insect larvae, resulting in significantly high levels of economic damage to crops.

Black mold, a disease caused by the fungus Pilgeriella anacardii Arx & Müller, affects cashews (Anacardium occidentale). Some cashew clones are more resistant to the pathogen; however, little is known about the chemical profile responsible for this trait. The investigation of volatile organic compounds (VOCs) from leaves of dwarf cashew clones resistant (BRS 226 and BRS 265) and susceptible (CCP 76 and BRS 189) to the pathogen was carried out. Leaves were collected during the months of disease incidence and decline (March to July 2019, Brazil), and VOCs were analyzed by gas chromatography-mass spectrometry (GC-MS) combined with chemometric tools. The GC-MS analysis tentatively identified 96 compounds. Partial least squares discriminant analysis (PLS-DA), orthogonal partial least squares discriminant analysis (OPLS DA), hierarchical cluster analysis (HCA), and ROC curves analysis were useful in dividing VOCs into distinct resistance and associated chemical susceptibility groups for different clones. The VOCs in the leaves of the resistant clones were identified as alcohols and aldehydes containing six carbons: (E)-hex-2-enal, hex-3-en-1-ol, (Z)-hex-2-en-1-ol, (E)-hex-2-en-1-ol, and hexan-1-ol. Moreover, α-pinene, pseudolimonene, α-phellandrene, β-myrcene, sylvestrene, β-cis-ocimene, methyl salicylate, myrtenol, α-copaene, γ-muurolene, germacrene D, valencene, and germacrene B were also detected in these samples and may be candidate chemical biomarkers for cashew resistance to P. anacardii.

Plant phenolic extracts are widely recognized as an important source of natural antioxidant substances and potential compounds for cosmetic formulations. This study aimed to evaluate the chemical profile, photoprotective and antioxidant activities of stem bark extract of Spondias purpurea L. (ciriguela) and its application in photoprotective formulations. Thirty phenolic constituents were annotated by ultra-performance liquid chromatography coupled with an electrospray ionization quadrupole time-of-flight mass spectrometry in mode negative (UPLC-QTOF-MS2). The stem bark extract antioxidant and chelation potential, expressed in half maximal inhibitory concentration (IC50), showed 6.25 and 352.22 µg mL−1, respectively. The phenolic extract was used as an active ingredient in six sunscreen formulations, with concentrations ranging from 0.2 to 10%. The ultraviolet (UV) protection properties of the formulations were evaluated by sun protection factor (SPF) values obtained in 0.2 mg mL−1 (0.495 to 2.27) and 2.0 mg mL−1 (2.29 to 15.87). The SPF value for the extract (14.37 and 26.16) was high, but there was a reduction in the base formulation. However, these results suggested that the bioactive extracted of stem bark of Spondias purpurea L. has interesting potential to reduce the damage caused by UV radiation and may be utilized as an active ingredient in a sunscreen formulation.

Cinnamic acid and its derivatives show a remarkable variety of biological activities and are often studied in search of the development of new and highly effective drugs. This work aims to synthesize, characterize and evaluate the cytotoxic activity of esters derived from cinnamic acid. Eighteen esters were synthesized through Steglich’s esterification, of which eleven were not reported in the literature. All compounds were fully characterized by Fourier transform infrared epectroscopy (FTIR), nuclear magnetic resonance (1H and 13C NMR) and high-resolution mass spectrometry (HRMS) data. The cytotoxic activity of esters obtained was evaluated using four human tumor cell lines: SNB-19 (astrocytoma), HCT-116 (colon carcinoma, human), PC3 (prostate) and HL60 (promyelocytic leukemia) through the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium (MTT) colorimetric assay. These studies showed that the compound 3-methoxybenzyl (E)-3-(4-methoxyphenyl)acrylate (12) is the most potent against HCT-116, PC3 and SBN-19 cells, with the lowest half maximal inhibitory concentration (IC50) value of 16.2 µM in the HCT-116 strain. The derivatives were obtained in good yields (76.6-95%), except for compounds 5-isopropyl-2-methylphenyl (E)-3-(3-hydroxy-4-methoxyphenyl)acrylate (17) (18.6%) and 2-isopropyl-5-methylphenyl (E)-3-(3-hydroxy-4-methoxyphenyl)acrylate (18) (15.5%).

Metabolomics, together with analytical methods and chemometric tools, point to new paths for selecting species that are resistant to pests and diseases. In this work, the forage palm species' metabolomic profile was investigated, and the relation between the chemical composition and resistance to Dactylopius opuntiae (carmine cochineal). The study was performed in cladodes of different non-susceptible cultivars (Nopalea cochenillifera (L.) Salm-Dyck, Opuntia stricta (Haw.), Nopalea cochenillifera and susceptible cultivar (Opuntia ficus-indica (L.) Mill.)). Metabolic profile showed 28 metabolites detected in the four species. From these total, 18 metabolites were annotated using UPLC-QTOF-MSE (ultra-performance liquid chromatography coupled with an electrospray ionization quadrupole time-of-flight mass spectrometry operating in MSE mode). By comparing the chemical profiles of non-susceptible and susceptible species through the application of chemometric tools, it was possible to obtain biomarkers (quercetin-3-O-2',6'-dirhamnosylglucoside, quercetin rhamnosyl dihexoside, and isorhamnetin-3-sophoroside-7-rhamnoside) that may be associated with resistance to carmine cochineal. Metabolomics based on UPLC-QTOF-MSE and chemometric allowed to establish the biomarkers knowledge of the resistance present in forage palm species. These results contribute to developing the initial understanding of flavonoids' role in the defense mechanisms of cactaceans and can be useful for application in breeding programs; it can increase the chances of success in creating new varieties of plants not susceptible to carmine cochineal.

The essential oil of Cymbopogon nardus, citronella, has been extensively studied. However, the chemical and biological properties of the ethanolic extract (EE) of C. nardus have not been evaluated. The aim of this study was to characterize the chemical composition of the EE of C. nardus and its active fraction (FrD). Moreover, the cytotoxic and antifungal properties of these extracts against Candida species with different resistance profiles to conventional drugs were evaluated. The compounds identified in EE were mono-C- and di-C-glycosyl flavones and phenylpropanoid glycosides. Phenylpropanoid glycosides were identified in FrD. EE showed antifungal activity, with minimum inhibitory concentration (MIC) values ranging from 62. 5 to 500 µg mL-1. FrD was more effective against C. glabrata, as evidenced by the lowest MIC value (15. 6 µg mL-1). EE inhibited yeast growth similar to amphotericin-B, as demonstrated by similar time-kill curves. EE inhibited C. albicans hyphae formation and mature biofilm of C. albicans, C. krusei and C. parapsilosis. The results of the chemical and biological analyses of EE and its fractions provided novel information and may contribute to control of infections caused by Candida species.

ABSTRACT The alternative use of plant essential oils as pesticide, repellent and/or antifeedant has been emphasized as an important tool in integrated pest management (IPM), promoted by the growing interest in methods that fulfil requirements of efficiency, safety, selectivity, technically feasible and environmentally safe. The objective of this work was to determine the pesticide potential of essential oils of rosemary pepper (Lippia origanoides Kunth; Verbenaceae), citronella (Cymbopogon winterianus Jowitt; Poaceae) and lemongrass (Cymbopogon citratus [DC.] Stapf; Poaceae) to control fall armyworm (Spodoptera frugiperda [J.E. Smith]; Lepidoptera: Noctuidae). Ovicidal, larvicidal and pupicidal activity was evaluated with five dosages (0.1%, 0.5%, 1.0%, 2.5% and 5.0% v/v) and a negative control (neutral detergent at 5.0% v/v). Unhatched eggs, mortality of caterpillars and non-emerged adults were recorded using ovicidal and pupicidal percentage, and median lethal doses (LD50) to caterpillars. The essential oils showed insecticidal activity on the egg, caterpillar 3rd instars and pupae stages, highlighting the rapid action and high mortality rates caused by the L. origanoides essential oil, which registered, at lowest dosage, average 97.8% ovicidal activity and 81.3% pupicidal activity; in addition to LD50 on 3rd instar caterpillars, 0.001% by exposure and 0.033% by topical application. Essential oils of C. citratus and C. winterianus required longer periods to act and caused significant mortality on the three stages, registering LD50 0.008% and 0.159% for exposure, and 1.151% and 1.348% for topical application, respectively. Citronella oil caused behavioral changes in caterpillars, which reflected flight and aggressiveness. Therefore, these three essential oils can be an alternative for management of fall armyworm, with the potential to hinder and reduce its biotic potential.

The collection time could be an important target for optimizing technological processes in obtaining essential oils of interest to agribusiness. The objective of this study was to distinguish the time for the collecting of Achillea millefolium L. leaves, which provides the highest yield and quality of the essential oil. Also, identify the type of polyethylene packaging and leaf storage period during the one-year period that would maintain the essential oil characteristics. Gas chromatography-mass spectrometry and chemometric studies were performed into two steps trial to detect changes in chemical profile induced by different conditions. First, an analysis of the leaf collection time was performed using chromatographic data from six different gathering times throughout the day. After determining the best time to collect from the leaves, the essential oil was extracted in five storage periods over a year. The highest oil content was observed in leaves harvested between 11 and 15 h, with a maximum of 39 min after 13 h. Therefore, it is recommended to perform extraction in the early afternoon. There was no significant statistical differentiation related to polyethylene packages. In addition, it is recommended that the essential oil can be stored without significant changes for up to six months.

Myracrodruon urundeuva (“aroeira-do-sertão”) is a species threatened with extinction due to anthropogenic exploitation. Phytochemical analysis of bark, branch and leaf extracts revealed the presence of several compounds such as flavonoids, phenols, tannins, quercetin derivatives and anacardic acids. Dereplication methodology was performed to tentatively identify 50 compounds analyzed by ultra-performance liquid chromatography coupled with an electrospray ionization quadrupole time-of-flight mass spectrometry operating in MSE mode (UPLC-QTOF-MSE). The extracts exhibited anti-tumor effect in cancer cells HCT-116 (colorectal), SF-295 (glioblastoma), HL-60 (leukemia), and RAJI (leukemia). Also, these results correlate with the principal component analysis (PCA) data that identified three distinct groups indicating, efficiently, metabolic differences between organs of M. urundeuva. Through discriminatory analysis of the orthogonal partial least squares (OPLS-DA), the variable of importance in the projection (VIP) and S-Plot, we were able to determine 30 potential biomarkers. The fingerprint of hydroethanolic extracts was correlated with the cytotoxicity assay and demonstrated a significant difference in the composition of plant extract.

Our study aimed at the development of a method based on ultra-performance liquid chromatography coupled to mass spectrometry (UPLC-ESI-MS) using selected ion monitoring for the simultaneous quantitation of the alkaloids galantamine, pseudolycorine, sanguinine and narciclasine in Amaryllidaceae species (Hippeastrum elegans, Habranthus cf. irwinianus, Hymenocallis littoralis and Griffinia nocturna). The alkaloids were extracted from dried and ground bulbs (100 mg) using liquid-liquid microextraction followed by solid phase extraction in cation exchange cartridges. The quantification method showed good linearity (correlation coefficient, R ≥ 0.9968) and selectivity for a run time of 10 min. All values were within the acceptable limits for recovery (87.5-96.2%), interday (coefficient of variation, CV%, 1.3-8.4%) and intraday precision (CV% 5.7-8.1%), except narciclasine (70%). The limits of detection and quantification were 5-100 and 20-350 ng mL-1, respectively. Our method demonstrated to be rapid, sensitive and reliable, therefore appropriate for being employed in the prospection of new source of galantamine and other alkaloids.

Agroindustrial residues, such as leaves of fruit plants, can be sources of bioactive molecules, thus adding value to co-products that are rarely explored in agroindustry. In this context, this study aimed to look into the phytochemical profiles in detail and to explore the antitumor potential of S. tuberosa and S. mombin leaves. We observed that, S. tuberosa leaf extract was cytotoxic in both tumor and healthy cells. S. mombin extracts selectively inhibited cell proliferation in the tumor cell line for prostate cancer (PC3) and did not significantly affect healthy cells. The metabolic profiles of the extracts were evaluated by gas chromatography coupled with mass spectrometry and twenty-three metabolites were identified. The correlation of metabolic profiles with cytotoxic tests indicated possible chemical markers that may be responsible for the inhibition of cell proliferation. This study revealed that the unexplored co-products in agroindustry may have great therapeutic potential, and therefore should be screened for biologically active compounds.

A method based on QuEChERS (quick, easy, cheap, effective, rugged, and safe) extraction and gas chromatography-mass spectrometry (GC-MS) detection was described for the pesticides dichlorvos, disulfoton, ethoprophos, parathion methyl, fenchlorphos, chlorpyrifos, azinphos methyl and prothiofos in sapodilla. For all compounds studies, it was observed a strong matrix effect showing the need to use matrix matched calibration strategy. Method was validated, and good linearity (R > 0.99) was obtained for all pesticides studied with limits of detection (LODs) and quantification (LOQs) ranging from 0.01 to 0.06 mg kg-1 and 0.03 to 0.2 mg kg-1, respectively. Recovery studies were performed at different levels (0.08, 0.10, 0.14, 0.20, 0.35 and 1.17 mg kg-1) and showed good results (between 70 and 120% with relative standard deviation (RSD) < 20%). A statistical test was applied to the coefficients of the analytical curves obtained in the sapodilla matrix. Analyses of commercial samples showed chlorpyrifos were detected in about 70 and 33% for fruit and pulps samples, respectively. It should be noted that chlorpyrifos is not permitted in sapodilla crops by ANVISA and EC guidelines.

Phytohormone analysis using liquid chromatography combined with tandem mass spectrometry is a very important tool for studies involving plant metabolism; however, this analysis can be affected by matrix composition. Although it is necessary to understand this effect to produce reliable results, it has been widely neglected in analyses of plant hormones. Leaf extracts from Bauhinia variegata were obtained by solvent extraction followed by solid-phase cleanup. The extract was analyzed with liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) in negative ionization mode, using the multiple reaction monitoring mode with two or three transitions to enhance analytical quality control. Although deuterated standards were used as surrogates, pronounced matrix effects were detected for phaseic acid (PA), abscisic acid (ABA) -glycosyl ester (ABA-GE) and dihydrophaseic acid (DPA), whereas ABA, neoPA and 7'-hydroxy-ABA (7'OHABA) showed negligible matrix effects. The method was validated using spiked samples and was applied to monitor ABA, PA, DPA, neoPA, 7´OHABA and ABA-GE on a daily basis. Analyte recoveries from spiked samples ranged from 67% to 87%. The highest leaf concentration of phytohormones was found at 2:00 pm and 5:00 pm, which represent typical day times related to the decrease of stomatal conductance. Despite the use of deuterated phytohormones as internal standards, we showed that the use of a calibration curve constructed with a matrix extract is mandatory for the reliable quantification of ABA and its metabolites, especially PA, ABA-GE and DPA. Daily variations in endogenous ABA leaf concentration were discussed.