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In this book, two of the primary sources of flavonoids, i.e. plants and fruits (oranges to be specific) are comprehensively reviewed, along with their flavonoid content and antioxidant activity (including action against COVID-19) and toxicity. Besides, a general overview of the analytical techniques for flavonoids determination in their natural sources is provided. Finally, a detailed description of flow techniques and their evolution, together with their role in the automation of the sample pretreatment and its combination with separation techniques, is also included.

Solanum ferruginium es una planta que crece en lugares perturbados como maleza, es de uso medicinal poco utilizada que presenta un gran potencial como fuente de antioxidantes debido a su alto contenido en polifenoles. Debido a esto se analizó el contenido de polifenoles, flavonoides, proantocianidinas y capacidad antioxidante in vitro (ensayo de captación de radicales DPPH) en hojas, tallos y planta completa de tres localidades (Las agujas, Parque el Nabo y Bosque la Primavera) de Zapopan, Jalisco, así como su toxicidad subcronica en hojas. En general se observó diferencia significativa (p£ 0,05), en las muestras de las tres localidades, las hojas presentaron el mayor contenido de polifenoles (15,3±0,7 a 22±0,4 mg expresado como equivalente de ácido gálico (EAG/g) en muestras de Parque el Nabo, flavonoides (7,8±0,3 a 13,3±0,3 mg EC/g) y proantocianidinas (3,4±0,1 a 4,2±0,05 mg expresado como equivalente de catequina (EC/g) en el Bosque la Primavera. La capacidad antioxidante fue similar en todas las muestras, con valores de 8,3 a 17 µg/mL de concentración media inhibitora (CI50). En la prueba toxicológica, los ratones no mostraron signos de toxicidad a ninguna dosis por efecto de la administración de la planta en estudio, por lo que la dosis letal media (DL50) es > 15 000 mg/kg de peso corporal. El contenido de polifenoles y actividad antioxidante en S. ferruginium sobre todo en hojas indican un alto potencial con propiedades farmacológicas además de su inocuidad, por lo que es importante realizar estudios de sus compuestos fenólicos individuales, antes de ser utilizada en farmacología.

Introduction: Medicinal plants contain essential phytochemicals and are used in traditional herbal therapy for ages. These phytochemicals are bioactive compounds and possess significant antibacterial and antifungal properties in addition to more biologically important potentials. The important bioactive phytochemicals include polyphenolic compounds and flavonoids. Arisaema tortuosum has been reported as a medicinal plant in literature and is used in treating many health issues. Aim: To investigate the phytochemical composition, total polyphenolic content (TPC), total flavonoid content (TFC), antioxidant capacity, and β-glucuronidase inhibition potential of the extract obtained from leaves of A. tortuosum. Methods: The phytochemical composition, TPC, TFC, and antioxidant capacity of the extract was analysed. β-glucuronidase inhibition assay was used to determine β-glucuronidase inhibition potential. The dose-dependent data was used to determine IC50 value of the extract. Results: The gas chromatography-mass spectroscopy (GC-MS) analysis identified the fifty-three components from the leaf extract that was observed to possess a significant volume of TPC and TFC. 1,1-diphenyl-2-picryl-hydrazyl (DPPH) assay revealed a IC50 value of 936 μg/mL indicating a high antioxidant activity of the extract. Conclusion: The study revealed the presence of many biologically important phytochemicals and a rich number of total polyphenols and flavonoids in leaf extract of A. tortuosum signifying the potential biomedical applications. Introducción: las plantas medicinales contienen compuestos fitoquímicos esenciales y se utilizan en la terapia herbolaria tradicional desde hace siglos. Estos fitoquímicos son compuestos bioactivos y poseen importantes propiedades antibacterianas y antifúngicas, además de potenciales efectos biológicamente más importantes. Los fitoquímicos bioactivos importantes incluyen compuestos polifenólicos y flavonoides. En la literatura se ha reportado a la Arisaema tortuosum como una planta medicinal y que se usa para tratar muchos problemas de salud. Objetivo: investigar la composición fitoquímica, el contenido polifenólico total (TPC), el contenido total de flavonoides (TFC), la capacidad antioxidante y el potencial de inhibición de la β-glucuronidasa del extracto obtenido de las hojas de A. tortuosum. Métodos: se analizó la composición fitoquímica, TPC, TFC y la capacidad antioxidante del extracto. Se utilizó el ensayo de inhibición de β-glucuronidasa para determinar el potencial de inhibición de este agente. Los datos dependientes de la dosis se usaron para determinar el valor IC50 del extracto. Resultados: el análisis de cromatografía de gases-espectroscopía de masas (GC-MS) permitió identificar cincuenta y tres componentes del extracto de hojas, que se observó que poseían un volumen significativo de TPC y TFC. El ensayo de 1,1-difenil-2-picril-hidrazilo (DPPH) reveló un valor IC50 de 936 μg/mL, lo que indica una alta actividad antioxidante del extracto. Conclusión: este estudio reveló la presencia de muchos compuestos fitoquímicos biológicamente importantes y una gran cantidad de polifenoles totales y flavonoides en el extracto de hoja de A. tortuosum, lo que significa aplicaciones biomédicas potenciales. Introdução: as plantas medicinais contêm compostos fitoquímicos essenciais e têm sido utilizadas na terapia tradicional à base de plantas há séculos. Esses fitoquímicos são compostos bioativos e possuem importantes propriedades antibacterianas e antifúngicas, além de efeitos potencialmente mais importantes biologicamente. Fitoquímicos bioativos importantes incluem compostos polifenólicos e flavonóides. Arisaema tortuosum tem sido relatada na literatura como planta medicinal e utilizada para o tratamento de diversos problemas de saúde. Objetivo: investigar a composição fitoquímica, teor de polifenóis totais (TPC), teor de flavonoides totais (TFC), capacidade antioxidante e potencial de inibição da β-glucuronidase do extrato obtido das folhas de A. tortuosum. Métodos: foram analisados a composição fitoquímica, TPC, TFC e a capacidade antioxidante do extrato. O ensaio de inibição da β-glucuronidase foi utilizado para determinar o potencial de inibição deste agente. Os dados dependentes da dose foram usados para determinar o valor IC50 do extrato. Resultados: a análise por cromatografia gasosa-espectroscopia de massa (GC-MS) permitiu a identificação de cinquenta e três componentes do extrato da folha, que apresentaram um volume significativo de TPC e TFC. O ensaio de 1,1-difenil-2-picril-hidrazil (DPPH) revelou um valor de IC50 de 936 μg/ mL, indicando uma alta atividade antioxidante do extrato. Conclusão: este estudo revelou a presença de muitos compostos fitoquímicos biologicamente importantes e uma grande quantidade de polifenóis totais e flavonóides no extrato da folha de A. tortuosum, significando potenciais aplicações biomédicas.

We studied the polyphenol (phenolic compounds and flavonoids) composition and allelopathic effects of Acacia melanoxylon R. Br. aerial foliage aqueous extract (0%, 25%, 50%, 75% and 100%) on the seedling growth and plant biomass of the general biotest species, lettuce (Lactuca sativa). Mean leaf fresh weight, leaf dry weight, root fresh weight and root dry weight were decreased following exposure to Acacia aerial foliage, flowers aqueous extract (AFE) and phyllodes aqueous extract (APE) after 6 days. The reduction in plant dry biomass was more than 50% following treatment with AFE. The decrease in mean root length was approximately 37.7% and 29.20% following treatment with Acacia flowers extract (AFE) at 75% and 100% concentration, respectively. Root dry weight of L. sativa was reduced by both flowers and phyllodes extract. The reduction of protein contents in lettuce leaves following Acacia foliage extract proved that both AFE and APE exhibit polyphenols that causes the toxicity which led to decrease in leaf protein contents. High-Performance Liquid Chromatography (HPLC) was employed to analyze the A. melanoxylon flowers and phyllodes. A total of 13 compounds (accounting for most abundant compounds in flowers and phyllodes) include different flavonoids and phenolic compounds. The phytochemical compounds detected were: Gallic acid, protocatechuic acid, p-hydroxybenzoic acid, p-hydroxybenzaldehyde, vanillic acid, syringic acid, p-coumaric acid, and ferulic acid. The major flavonoid compounds identified include rutin, luteolin, apigenin, and catechin. Allelopathic effects of flower and phyllodes extracts from A. melanoxylon may be due to the presence of above compounds identified by HPLC analysis.

Apigenin, a member of the flavone subclass of flavonoids, has long been considered to have various biological activities. Its glucosides, in particular, have been reported to have higher water solubility, increased chemical stability, and enhanced biological activities. Here, the synthesis of apigenin glucosides by the in vitro glucosylation reaction was successfully performed using a UDP-glucosyltransferase YjiC, from Bacillus licheniformis DSM 13. The glucosylation has been confirmed at the phenolic groups of C-4 and C-7 positions ensuing apigenin 4-O-glucoside, apigenin 7-O-glucoside and apigenin 4,7-O-diglucoside as the products leaving the C-5 position unglucosylated. The position of glucosylation and the chemical structures of glucosides were elucidated by liquid chromatography/mass spectroscopy and nuclear magnetic resonance spectroscopy. The parameters such as pH, UDP glucose concentration and time of incubation were also analyzed during this study.

Under greenhouse conditions, the resistance of 18 different genotypes of flax to powdery mildew was evaluated. To investigate genetic diversity and identify the molecular and biochemical markers linked to powdery mildew resistance in the tested genotypes, two molecular marker systems—start codon targeted (SCoT) and inter-simple sequence repeat (ISSR)—as well as a biochemical marker (protein profiles, antioxidant enzyme activity, and secondary metabolites) were used. Based on the results, the genotypes were classified into four categories: highly susceptible, susceptible, moderately susceptible, and moderately resistant. The genotypes differed significantly in powdery mildew severity: Polk had a severity of 92.03% and Leona had a severity of 18.10%. Compared to the other genotypes, the moderately resistant genotypes had higher levels of flavonoids, antioxidant enzymes, phenolics, and straw yield; nevertheless, their hydrogen peroxide and malondialdehyde levels were lower. Protein profiles revealed 93.75% polymorphism, although the ISSR marker displayed more polymorphism (78.4%) than the SCoT marker (59.7%). Specific molecular and biochemical markers associated with powdery mildew resistance were identified. The 18 genotypes of flax were divided into two major clusters by the dendrogram based on the combined data of molecular markers. The first main cluster included Leona (genotype number 7), considered moderate resistance to powdery mildew and a separate phenetic line. The second main cluster included the other 17 genotypes, which are grouped together in a sub-cluster. This means that, besides SCoT, ISSR markers can be a useful supplementary technique for molecular flax characterization and for identifying genetic associations between flax genotypes under powdery mildew infection.

An 80% methanolic extract of Rhus verniciflua Stokes bark showed significant anti-viral activity against fish pathogenic infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV) in a cell-based assay measuring virus-induced cytopathic effect (CPE). Activity-guided fractionation and isolation for the 80% methanolic extract of R. verniciflua yielded the most active ethyl acetate fraction, and methyl gallate (1) and four flavonoids: fustin (2), fisetin (3), butin (4) and sulfuretin (5). Among them, fisetin (3) exhibited high antiviral activities against both IHNV and VHSV showing EC∧50 values of 27.1 and 33.3 μM with selective indices (SI = CC∧50/EC∧50) more than 15, respectively. Fustin (2) and sulfuretin (5) displayed significant antiviral activities showing EC∧50 values of 91.2-197.3 μM against IHNV and VHSV. In addition, the antiviral activity of fisetin against IHNV and VHSV occurred up to 5 hr post-infection and was not associated with direct virucidal effects in a timed addition study using a plaque reduction assay. These results suggested that the bark of R. verniciflua and isolated flavonoids have significant anti-viral activity against IHNV and VHSV, and also have potential to be used as anti-viral therapeutics against fish viral diseases.

Pleurotus eous is a pink edible oyster mushroom cultivated in Coimbatore. The aim of present study was to evaluate antioxidant potential of ethyl acetate, methanol, and hot water extracts from P. eous using various in vitro ROS/RNS generated chemical and biological models. Results demonstrated that the graded-dose (2–50 mg/mL) of various extracts markedly scavenged the DPPH, ABTS, and hydroxyl radicals, and showed metal chelating ability, reducing capacity in Fe3+/ferricyanide complex, and ferric reducing antioxidant power assays. In biological models, different extracts were found to inhibit lipid peroxidation in brain and liver homogenate. Total antioxidant activity was high in methanolic extract. Hot water extract showed higher amount of total phenol and methanolic extract showed higher amount of flavonoid as compared to other extracts. Based on EC50 values it can be concluded that the various extracts from P. eous were good in antioxidant properties with methanol and ethyl acetate extracts being more effective. The results of this study showed that various extracts could serve as natural antioxidants owing to their significant antioxidant activity.

We described the in vitro antioxidant and antimicrobial activities of ethanol, acetone, and water extracts of beet root pomace. Total contents of phenolics (316.30-564.50 mg GAE/g of dry extract), flavonoids (316.30-564.50 mg RE/g of dry extract), betacyanins (18.78-24.18 mg/g of dry extract), and betaxanthins (11.19-22.90 mg/g of dry extract) after solid-phase extraction were determined spectrophotometrically. The antioxidant activity was determined by measuring the reducing power and DPPH scavenging activity by spectrometric method, and hydroxyl and superoxide anion radical scavenging activity by ESR spectroscopy. In general, the reducing power of all the beet root pomace extracts increased with increasing concentrations. The DPPH-free radical scavenging activity of the extracts, expressed as EC50, ranged from 0.133 mg/mL to 0.275 mg/mL. Significant correlation was observed between all phytochemical components and scavenging activity. Ethanol extract (0.5 mg/mL) completely eliminated hydroxyl radical which had been generated in Fenton system, while the same concentration of this extract scavenged 75% of superoxide anion radicals. In antibacterial tests, Staphylococcus aureus and Bacillus cereus showed higher susceptibility than Escherichia coli and Pseudomonas aeruginosa.

The objective of the present study was to evaluate and compare the phenolic content (total phenols and flavonoids) and the antioxidant capacity of seventeen craft beers produced in Mexico. The results showed a considerable amount of content of total phenols and flavonoids in the styles Coffee Stout Imperial (362 mg GAE/L and 75 mg QE/L) and Stout Ale (299 mg GAE/L and 40.92 mg QE/L), followed by American Brown Ale (271 mg GAE/L and 27.35 mg QE/L), Brown Ale (211 mg GAE/L and 24.20 mg QE/L), and Dark (328 mg GAE/L and 20.94 mg QE/L). In contrast, the American Wheat Ale (65 mg GAE/L and 2.91 mg QE/L), Lager (102 mg GAE/L and 3.56 mg QE/L), and IPA (103.73 mg GAE/L and 3.75 mg QE/L) showed the lowest content of phenols and flavonoids. The antioxidant capacity is detected in all the beers evaluated, mainly in the Coffee Stout Imperial style beer (80%). In this study, different contents of total carbohydrates were observed according to the type of beer, with Coffee Stout Imperial, American Brown Ale, and Brown being the ones that presented the highest content of carbohydrates (24.25, 15.55, and 15.36 g/L, respectively) followed by the type of Pilsner beer. (13.60g/L). Finally, our study showed that the different styles of beers in Mexico presented different contents of polyphenols (phenols and flavonoids), antioxidant capacity, and carbohydrates. However, the Coffee Stout Imperial style beer showed the highest values ​​of the aforementioned parameters. Future studies should be carried out in the evaluation of the biological activity of the different styles of beers on nutritional parameters in the Mexican population.