Comprehensive assessment of clean-up strategies for optimizing an analytical multi-method to determine pesticides and mycotoxins in Brazilian medicinal herbs using QuEChERS-LC-TQ-MS/MS.

The use of medicinal herbs has increased significantly. However, the presence of pesticide residues and mycotoxins in medicinal herbs has generated constant discussion and concern among regulatory agencies. Developing and validating an analytical method for determining pesticides and mycotoxins in medicinal plants is challenging due to the naturally occurring substances in these plants. The purpose of this work was to develop and to optimize a sensitive, accurate, precise, effective QuEChERS method for simultaneous determination of over 160 pesticide and mycotoxin residues in complex medicinal plant matrices using LC-TQ-MS/MS. A comprehensive comparison of clean-up procedures and other parameters was conducted to achieve this goal. The validation procedure was performed according to SANTE 11312/2021. More polar analytes, such as acephate, methamidophos and omethoate, presented a higher negative matrix effect in both Melissa officinalis L. and Malva sylvestris L. However, other molecules, such as spirodiclofen, showed a 24% signal enhancement in M. officinalis and a 46% signal suppression in M. sylvestris, indicating that a representative matrix-matched calibration would lead to inaccurate quantification of the analyte. Accuracy and precision were satisfactory according to SANTE 11312/2021 for 157 pesticide residues and mycotoxins in M. officinalis and for 152 molecules in M. sylvestris. LOQs at 10 µg kg-1 were achieved for 117 pesticides in M. officinalis and 99 pesticides in M. sylvestris. Among the mycotoxins, all four aflatoxins (B1, B2, G1 and G2) presented LOQs of 5 µg kg-1, and ochratoxin A had an LOQ of 10 µg kg-1 in M. officinalis. The same LOQ values were shown for these mycotoxins in M. sylvestris, except for aflatoxin B2 and ochratoxin A, which had LOQs of 20 µg kg-1. Moreover, in Southern Brazil, there has been no previous study on mycotoxin and pesticide contamination in medicinal herbs. Therefore, the application of this method was assessed through the analysis of forty-two real samples. Imidacloprid was found in M. officinalis, and methyl pirimiphos was found in M. sylvestris. The proposed method not only serves as a helpful tool for routine monitoring but also offers a basis for further research on risk assessment and control in food safety.

Probiotic Yogurt Supplemented with Lactococcus lactis R7 and Red Guava Extract: Bioaccessibility of Phenolic Compounds and Influence in Antioxidant Activity and Action of Alpha-amylase and Alpha-glucosidase Enzymes.

The industry has increasingly explored the development of foods with functional properties, where supplementation with probiotics and bioactive compounds has gained prominence. In this context, the study aimed to evaluate the influence of in vitro biological digestion on the content of phenolic compounds, antioxidant activity, and inhibition of α-amylase and α-glucosidase activities of probiotic yogurt supplemented with the lactic acid bacteria Lactococcus lactis R7 and red guava extract (Psidium cattleianum). A yogurt containing L. lactis R7 (0.1%) and red guava extract (4%) was characterized for the content of phenolic compounds, antioxidant activity, and potential for inhibition of digestive enzymes after a simulated in vitro digestion process. After digestion, the caffeic and hydroxybenzoic acids remained, and sinapic acid only in the last digestive phase. Antioxidant activity decreased during digestion by 28.93, 53.60, and 27.97% for DPPH, nitric oxide and hydroxyl radicals, respectively, and the inhibition of the α-amylase enzyme decreased only 4.01% after the digestion process. α-glucosidase was more efficient in intestinal digestion, demonstrating an increase of almost 50% in probiotic yogurt with red guava extract before digestion. Possibly, the phenolics change their conformation during digestion, generating new compounds, reducing antioxidant activity, and increasing the inhibitory activity of α-glucosidase digestive enzymes. It was concluded that the probiotic yogurt formulation supplemented with red guava extract could interfere with the concentration of phenolic compounds and the formation of new compounds, suggesting a positive and effective inhibition of the digestive enzymes, even after the digestive process.

Lime and orange essential oils and d-limonene as a potential COVID-19 inhibitor: Computational, in chemico, and cytotoxicity analysis.

The COVID-19 pandemic has substantially impacted the world health systems, causing public health concerns, and the search for new compounds with antiviral activity is of extreme interest. Natural molecules with bioactive potential are a trend, with essential oils (Eos) being the focus of recent studies. Thus, this study evaluates in chemico the d-limonene inhibitory activities in the viral genome of SARS-CoV-2 and analyzes the cytotoxic potential and safety profile of d-limonene and lime and orange EOs with a high content of d-limonene. The EOs were extracted and characterized, and the in chemico computational analysis for the determination as a potential anti-SARS-CoV-2 was performed with d-limonene, the major compound in EOs. The cytotoxicity analysis of EOs and d-limonene was carried out with MRC-5 and HaCaT, and the preliminary safety profile was also evaluated by the HET-CAM assay. d-limonene was suggested as a promising compound for anti-SARS-CoV-2 research, since the molecule does not provide mutagenic and cytotoxic fragments, and does not have irritating potential when diluted, in addition to having favorable pharmacokinetic characteristics, through in chemico analysis. Collectively, the results reveal the antiviral potential of lime and orange EOs, as well as their major compound. In this sense, further studies should be conducted to understand the antiviral mechanisms.

Chemical, antioxidant and cytotoxic profile of hydroalcoholic extracts of plants from Southern Brazil and their activity against pathogenic fungi isolated from dogs and cats with sensitivity and resistance to conventional antifungals.

We aimed to evaluate the chemical, antioxidant, cytotoxic, and antifungal activities of hydroalcoholic extracts of native plants from Southern Brazil: Schinus terebinthifolia (SCH), Persicaria hydropiperoides (PER), Eugenia uniflora (EUG) and Equisetum hyemale (EQU). Ethyl gallate, quercetin, and quinic acid were prevalent compounds identified by LC-MS. For total phenolic/flavonoid contents and the antioxidant potential against ABTS/DPPH radicals, the ascending order was EQU < PER < EUG < SCH. All extracts were low cytotoxic and kept a high Vero cell viability (>75%) at concentrations up to 12.5 mg/mL (MTT assay). By M38-A2/M27-A3 (CLSI) against 68 clinical isolates of animals and strains of Malassezia pachydermatis, Sporothrix brasiliensis, Microsporum canis, Microsporum gypseum and Trichophyton mentagrophytes, all extracts (MIC/MFC ≤3.13-100 mg/mL) were active, except EUG. SCH inhibited and killed S. brasiliensis (MIC/MFC50/90 3.12-12.5 mg/mL) and dermatophytes (MIC/MFC 6.25-25 mg/mL) resistant to ketoconazole and itraconazole. These findings support the promising use of the selected plant extracts as antifungal agents.

Antimicrobial and cytotoxic activity to human colon adenocarcinoma cell lines (HT-29) potential of olive oil extraction residue.

In the Olive drupe (Olea europaea L.) oil extraction process, 80% of the volume generated is waste (bagasse). Advancing the expansion of the olive oil market, it is necessary to develop alternatives that, in addition to adding value to industrial waste, also reduce possible environmental damage. Our study aimed to understand the antimicrobial and Cytotoxic activity potential of the residues from the extraction of olive oil from the blend of the varieties Arbequina and Arbosana. The extract shows cytotoxic activity, inhibiting about 75% of cancer cells in the human colon at a concentration of 0.15 mg of Gallic Acid equivalent (GAE)/mL. The effectiveness of the extract against microorganisms often associated with foodborne diseases and food decomposition has also been discovered, without compromising the microorganisms responsible for fermentation. Thus, this study provides future perspectives for the use of active ingredients extracted from the residue from the extraction of olive oil.

Multiresistant bacteria isolated from domestic and wild animals with skin lesions were susceptible to native plants from Southern Brazil.

We evaluated the chemical composition, toxicity, and antibacterial activity of Schinus terebinthifolia (SCH), Eugenia uniflora (EUG), Persicaria hydropiperoides (PER), Equisetum hyemale (EQU), Solidago chilensis (SOL), and Baccharis trimera (BRA). These plants were tested (7.5-0.01 mg/mL) against Gram-positive (G+; n = 32) and Gram-negative (G-; n = 26) isolates from animals (M07-A9, CLSI). Antibiogram (disk diffusion), chromatographic analysis (UPLC), and toxicity assay (HET-CAM) were also performed. A high incidence of resistance was noted, in which 18.4% (07/38) of G+ (Staphylococcus intermedius/Enterococcus faecium) and 17.7% (06/34) of G- (Pseudomonas aeruginosa/Escherichia coli/Proteus mirabilis) were multidrug-resistant. All bacteria were sensitive (MIC50) to SCH (both 3.75 mg/mL), EUG (3.75 mg/mL and 7.5 mg/mL, respectively) and PER (both 7.5 mg/mL). SCH/EUG/PER highlighted as antibacterial, probably due to the major compounds (ethyl gallate, quinic acid, quercetin). These extracts showed normal embryonic development (SCH/EUG: 7.5-0.94 mg/mL). These findings highlighted the promising use of native plants for therapeutic purposes.

Bioactive content of six passion fruit genotypes cultivated in southern Brazil

Passion fruit production in Brazil is concentrated in tropical regions, however, recently production has expanded to regions with temperate climate. Cultivar performance in different climate and soil can lead to variation in the contents of bioactive compounds in the juice and rind of the fruit. This study characterized the bioactive content of passion fruit rind and juice of six passion fruit genotypes ('Catarina Roxo', 'Catarina', 'Urussanga', 'BRS Gigante Amarelo', 'BRS Rubi do Cerrado', and 'BRS Sol do Cerrado') cultivated in southern Brazil. Phenolic, flavonoid, anthocyanin, carotenoid, and ascorbic acid contents and the antioxidant potential of each fruit portion from each genotype were evaluated. Fruit composition varied with genotype and evaluated tissue. Bioactive compound contents were greatest in the rind of 'Urussanga' and 'Catarina', whereas the juice of 'BRS Sol do Cerrado' and 'Catarina Roxo' had the greatest bioactive content. Given cultivar and tissue variability for bioactive content, postharvest use of passion fruit will determine the choice of cultivar to obtain a product with maximum bioactive compounds. Passion fruit genotypes evaluated in this study have shown to be potential sources of bioactive compounds. Fruit rind has the potential to be explored in the scientific and technological scope, due to its high amounts of phenolic compounds and antioxidant activity.

Jabuticaba [Plinia peruviana (Poir.) Govaerts]: a Brazilian fruit with a promising application against itraconazole-susceptible and -resistant Sporothrix brasiliensis.

Hydroalcoholic extract from Jabuticaba peels was evaluated for the chemical profile, antioxidant potential, cytotoxicity, and anti-Sporothrix brasiliensis activities against both dimorphic phases. Higher phenolic content (14.91 ± 0.97) compared to flavonoid (2.05 ± 1.00) associated with the ellagic acid (1.93 ± 0.03; LC-MS), and a good scavenging ability against ABST and DPPH radicals was noted. On MDBK cells, elevated cell viability (>90%) was demonstrated between 0.39 and 0.097 mg/ml (MTT assay). Mycelial (CLSI M38-A2) and yeast (CLSI M27-A3) phases of 18 isolates of Sporothrix brasiliensis from cats (n = 8), dogs (n = 8) and humans (n = 2) were used. They were identified itraconazole-susceptible and itraconazole-resistant isolates in both phases, which were all inhibited (MIC of ≤1.56-6.25 mg/ml for both phases) and killed (MFC of ≤1.56-12.5 mg/ml for mycelial; ≤1.56-50 mg/ml for yeast) by Jabuticaba. For the first time, these findings highlighted the potential usefulness of hydroalcoholic extract from Jabuticaba peel as a promising antifungal against sporotrichosis.

Hypolipidemic and anti-inflammatory properties of phenolic rich Butia odorata fruit extract: potential involvement of paraoxonase activity.

Aim: This study investigated the effects of polar Butia odorata fruit extract on metabolic, inflammatory, and oxidative stress parameters in rats submitted to a hyperlipidaemia condition induced by tyloxapol.Methods: Animals were divided into 3 groups: saline, saline plus tyloxapol, and B. odorata extract plus tyloxapol. Animals were treated for 15 days with a saline solution or B. odorata fruit extract and after hyperlipidaemia was induced by tyloxapol.Results: Treatment with B. odorata extract reduced serum triglyceride, total cholesterol, C-reactive protein, and adenosine deaminase and butyrylcholinesterase activities when compared to the tyloxapol group. HDL-cholesterol and paraoxonase 1 activity were higher in B. odorata extract treated animals when compared to tyloxapol-treated animals. No differences were observed in hepatic oxidative stress parameters. Phenolic compounds present in B. odorata fruit extract were identified and quantified by LC-MS/MS.Conclusion: These findings indicated that phenolic rich B. odorata extract has hypolipidemic and anti-inflammatory effects in hyperlipidemic rats.

Effects of drying temperature and long-term storage conditions on black rice phenolic compounds.

This study aimed to investigate the influence of drying temperatures (20, 40, 60, 80, and 100 °C) followed by 12 months-storage under normal-atmosphere (conventional), nitrogen-atmosphere, and vacuum-atmosphere on black rice phenolics. Increase in drying temperature reduced the total content of free-phenolics, free-flavonoids, and anthocyanins. An increase in bound phenolics extractability was observed at 60 and 80 °C, suggesting some extent of phenolics polymerization and complexation at these drying temperatures. The free fraction of ferulic, caffeic, p-coumaric, and gallic acids were the most thermally unstable compounds. After storage, no reductions were observed only for free caffeic and protocatechuic acids. For total flavonoids, an increase in the free fraction and reduction of the bound fraction were observed in the grains stored under conventional- and vacuum-atmosphere. A reduction of total phenolics occurred regardless of the storage condition. However, storage under nitrogen-atmosphere is the most recommendable condition for maintenance of phenolics in black rice.

Discrimination of genotype and geographical origin of black rice grown in Brazil by LC-MS analysis of phenolics.

Physicochemical properties, cooking time, and phenolics profile of two black rice genotypes grown at six different locations in Brazil were determined. The cultivar IAC 600 and the elite-line AE 153045 were used. The main growing locations for black rice were considered, as follows: Alegrete (ALG), Capão do Leão (CPL), Guaratinguetá (GUA), Roseira (ROS), Santa Vitória do Palmar (SVP), and Taubaté (TBT). Principal component analysis (PCA) and supervised partial least squares-discriminant analysis (PLS-DA) from liquid chromatography-mass spectrometry (LC-MS) data sets showed distinction among genotypes and locations. Quercetin-3-O-glucoside and vanillic acid were the most relevant compounds for discriminating genotypes. SVP location provided the most distinctive black rice, with greater total phenolics content. Characteristics of black rice from SVP location were associated to effects of latitude and wind conditions. Hesperetin, vanillic acid, quercetion-3-O-glucoside, and p-coumaric acid were the most relevant compounds for discriminating locations.

Hydrothermal treatment of maize: Changes in physical, chemical, and functional properties.

The objective of this work was to assess the effects of a traditional parboiling treatment on physical, chemical and functional properties of yellow maize kernels. For this, maize kernels were subjected to the three main stages of a traditional parboiling process (soaking, steaming, and drying) at different moisture contents (15%, 25%, or 35%), and different pressure steaming times (0, 15, or 30 min). Kernels were evaluated for physical and chemical changes, while manually generated endosperm fractions were further evaluated for nutritional and functional changes. The parboiling process negatively altered the maize kernels properties by increasing the number of kernels with burst pericarp and decreasing the total carotenoid content in the endosperm by 42%. However, the most intense conditions (35% moisture and 30 min steam) lowered the number of broken kernels by 41%, and the number of stress cracks by 36%. Results also demonstrated that soaking enhanced the nutritional value of soaked yellow maize by increasing the thiamine content and the bound phenolic content in the endosperm fraction up to 102%. The proper implementation of this hydrothermal treatment could lead to significant enhancements in nutritional and functionality of maize products.

Optimized Camellia sinensis var. sinensis, Ilex paraguariensis, and Aspalathus linearis blend presents high antioxidant and antiproliferative activities in a beverage model.

A statistical optimization study was conducted to obtain a tea containing fermented rooibos (Aspalathus linearis), white tea (Camellia sinensis var. sinensis), and roasted mate (Ilex paraguariensis). An optimal combination of these species was proposed. This optimized tea inhibited 64% the lipoperoxidation in vitro and presented a high phenolic content, especially kaempferol, (+)-catechin, (-)-epicatechin, rutin, (-)-epigallocatechin, and (-)-epicatechin-2-O-gallate. Indeed, the antioxidant effect was confirmed by decreasing 30% the reactive oxygen species generation in human hepatoma carcinoma cells (HepG2, 100 and 240 µg/mL). In the cell viability assay, the GI50 for human colorectal adenocarcinoma epithelial cells (Caco-2) was about 547 µg/mL and 481 µg/mL for HepG2. The pasteurization process (65 °C/30 min) did not affect the total phenolic content and antioxidant activity of the optimized tea formulation. The sensory test indicated an acceptability index of 78%, showing that the analytical approach adopted was feasible to develop a phenolic-rich beverage.

Quality of black beans as a function of long-term storage and moldy development: Chemical and functional properties of flour and isolated protein.

The aim of this study was to evaluate the effects of moisture content and storage temperature on the percentage of moldy and fermented beans, mycotoxins levels, phenolic acids content, pasting properties of whole flour, as well as functional and thermal properties of protein isolates from black beans stored for 12 months. Beans stored under 14%/32 °C exhibited 16% of fermented grains, while at 17%/25 °C (42.3%) and 17%/32 °C (93.5%) of moldy plus fermented grains, named drastic conditions (DC). Mycotoxins were not present in grains from all storage conditions. Reduction of gallic, caffeic, and p-hydroxybenzoic acid contents, and increase of sinapic acid were observed in DC. Reduction of peak, final, and setback viscosities of bean flours in DC indicate the application in refrigerated and frozen products. The increase in foaming and reduction in foam degradation of the proteins highlights their use in beverages where the foam is an important factor.

Polar Origanum vulgare (Lamiaceae) extracts with antifungal potential against Sporothrix brasiliensis.

Oregano (Origanum vulgare) has anti-Sporothrix spp. activity, including against strains that are resistant to antifungal drugs. As the studies are limited to the essential oil, the aim of this study was to evaluate the chemical, antioxidant and cytotoxic properties of polar oregano extracts and their anti-Sporothrix brasiliensis activity. Aerial plant parts were used in the preparation of 10 min (INF10) and 60 min (INF60) infusions, a decoction (DEC) and a hydroalcoholic extract (HAE). Six phenolic acids and four flavonoids were identified and quantified through liquid-chromatography (LC-MS). Extracts in increasing order of total phenolic and flavonoid contents were HAE 40 mg/ml for the other extracts. This is the first report of oregano extracts showing antifungal activity against S. brasiliensis. Its use in the treatment of sporotrichosis may be considered upon toxicity and safe-use conditions are tested.

Effects of Organic and Conventional Cropping Systems on Technological Properties and Phenolic Compounds of Freshly Harvested and Stored Rice.

This study compared the physicochemical and technological properties of the IRGA 410 rice cultivar, obtained from organic and conventional cropping systems, and showed its susceptibility to changes during storage at 0, 6, and 12 mo. The rice conventional cropping system exhibited greater protein, lipids, and ash levels, and a head rice yield. However, organic rice presented greater total carbohydrates, soluble protein, amylose content, 33% greater free phenolics, and phenolic acids, but exhibited a greater percentage of stained grains during the storage period. The free phenolic content of cooked rice was lower than the free phenolic content of the raw rice. By Liquid chromatography with mass spectrometer (LC-MS) were identified p-coumaric and ferulic acids in both fractions (free and bound). The content of p-coumaric acid and ferulic acid in bound fraction was higher in organic brown rice than in conventional brown rice. At 6 and 12 mo of storage, the main fungi found were Aspergillus sp. and Penicillium sp. Prior to storage, the Bipolaris sp. fungi was identified only in organic rice. For conventional rice, the infestation level by Aspergillus sp. increased from 3% to 70% at the 6th mo of storage. In addition to the advantage of organic rice being free of agrochemicals, this study revealed that natural plant defense compounds could be produced when the rice was subjected to more biotic and abiotic stresses. However, some disadvantages were observed, such as lower protein content and a greater percentage of soluble protein, which favored the breaking of rice in processing, and a greater percentage of grain stained before and during storage. PRACTICAL APPLICATION: The organic and conventional cropping systems affect the physicochemical and technological properties of rice grains, which is one of the main cereals grown and consumed in the world. This study shows the advantages and disadvantages of the cropping system in grain properties that are in the interest of both consumers and processing industries.

Stability of bioactive compounds in butiá (Butia odorata) fruit pulp and nectar.

Butia odorata is a palm tree native to southern Brazil whose fruit (known as butiá) and leaves are used to make many food products and crafts. Butiá contain several biologically active compounds with potential health benefits. However, processing conditions can alter quality attributes including bioactive compound content. This study evaluated the stability of bioactive compounds in butiá pulp upon pasteurization, during 12months of frozen storage, and in butiá nectar after a 3-month storage period. Pulp pasteurization resulted in a reduction in phenolic, flavonoid, carotenoid, and ascorbic acid contents. After a 12-month frozen storage period, flavonoid, phenolic, and ascorbic acid contents decreased while carotenoid content remained unaltered. Carotenoid, ascorbic acid, and phenolic contents were unaffected by the 3-month storage of butiá nectar; however, flavonoid content and antioxidant potential were reduced. Despite bioactive compound degradation upon heat treatment and storage, butiá nectar remained rich in phenolics, especially (-)-epicatechin, rutin, and (+)-catechin.

Foliar Desiccators Glyphosate, Carfentrazone, and Paraquat Affect the Technological and Chemical Properties of Cowpea Grains.

The effects of the use of glyphosate (GLY), glyphosate plus carfentrazone (GLY/CAR), and paraquat (PAR) as plant desiccators on the technological and chemical properties of cowpea grains were investigated. All studied desiccants provided lower cooking time to freshly harvested cowpea. However, the coat color of PAR- and GLY/CAR-treated cowpea was reddish in comparison to the control treatment. Principal component analysis (PCA) from liquid chromatography-mass spectrometry (LC-MS) data sets showed a clear distinction among cowpea from the different treatments. Catechin-3-glucoside and epicatechin significantly contributed for discriminating GLY-treated cowpea, while citric acid was responsible for discriminating GLY/CAR-treated cowpea. Quercetin derivative and gluconic acid were responsible for discriminating control treatment. Residual glyphosate and paraquat content was higher than the maximum limits allowed by Codex Alimentarius and the European Union Commission. Improvements in the technological and chemical properties of cowpea may not be overlapped by the risks that those desiccants exhibit when exceeding the maximum limits of tolerance in food.

Chemical composition and cytotoxicity of extracts of marjoram and rosemary and their activity against Sporothrix brasiliensis.

PURPOSE: Motivated by increasing reports of antifungal resistance in human and animal sporotrichosis, this study evaluated the chemical composition, cytotoxicity and anti-Sporothrix brasiliensis activity of extracts of marjoram (Origanum majorana) and rosemary (Rosmarinus officinalis). METHODOLOGY: Ten (INF10) and 60 min (INF60) infusions, a decoction and a hydroalcoholic extract (HAE, 70 %) were prepared from both plants (10 % w/v). The extract composition was analysed by liquid chromatography/mass spectrometry and the cytotoxicity was evaluated using a colorimetric assay in canine and feline kidney cells. Using a broth microdilution assay (CLSI M38-A2) adapted to the extracts, 30 Sporothrix brasiliensis isolates from dogs, cats and humans, and one Sporothrix schenckii were tested.Results/Key findings. The predominant phenolic compounds found in all extracts were 4-hydroxybenzoic acid, caffeic acid and chlorogenic acid. Luteolin was also one of the predominant compounds, but only in the HAE of marjoram. Extracts of marjoram maintained cell viability in concentrations up to 2.5 mg ml-1 for the feline cell line and up to 10 mg ml-1 for the canine cell line, whereas in rosemary, the cell viability for both kidney lines was maintained with concentrations up to 5 mg ml-1. The activity of rosemary extracts was low or absent. Among the marjoram extracts, HAE was highlighted and had fungistatic activity against Sporothrix brasiliensis (MIC5040 mg ml-1), including in all itraconazole-resistant isolates. S. schenckiisensu stricto was sensitive to marjoram extracts (MIC/MFC ≤5 mg ml-1), with the exception of INF10. CONCLUSION: These findings support the potential usefulness of the HAE of marjoram in the treatment of sporotrichosis.