Iris Catiana Zampini

Singlet oxygen quenching and radical scavenging capacities of structurally-related flavonoids present in Zuccagnia punctata Cav.

The singlet oxygen (1O2) quenching and free radical (DPPH•, ABTS• + and O2• −) scavenging ability of three structurally-related flavonoids (7-hydroxyflavanone HF, 2′,4′-dihydroxychalcone DHC and 3,7-dihydroxyflavone DHF) present in the Argentinean native shrub Zuccagnia punctata Cav. were studied in solution by combining electrochemical and kinetic measurements, mass spectroscopy, end-point antioxidant assays and computational calculations. The results showed that the antioxidant properties of these flavonoids depend on several factors, such as their electron- and hydrogen atom donor capacity, the ionization degree of the more acidic group, solvatation effects and electrostatic interactions with the oxidant species. The theoretical calculations for both the gas and solution phases at the B3LYP level of theory for the Osanger reaction field model agreed with the experimental findings, thus supporting the characterization of the antioxidant mechanism of the Z. punctata flavonoids.

Polyphenols rich fraction from Geoffroea decorticans fruits flour affects key enzymes involved in metabolic syndrome, oxidative stress and inflammatory process

Geoffroea decorticans (chañar), is widely distributed throughout Northwestern Argentina. Its fruit is consumed as flour, arrope or hydroalcoholic beverage. The chañar fruits flour was obtained and 39 phenolic compounds were tentatively identified by HPLC-MS/MS(n). The compounds comprised caffeic acid glycosides, simple phenolics (protocatechuic acid and vanillic acid), a glycoside of vanillic acid, p-coumaric acid and its phenethyl ester as well as free and glycosylated flavonoids. The polyphenols enriched extract with and without gastroduodenal digestion inhibited enzymes associated with metabolic syndrome, including α-amylase, α-glucosidase, lipase and hydroxyl methyl glutaryl CoA reductase. The polyphenolic extract exhibited antioxidant activity by different mechanisms and inhibited the pro-inflammatory enzymes (ciclooxygenase, lipoxygenase and phospholipase A2). The polyphenolic extract did not showed mutagenic effect by Ames test against Salmonella typhimurium TA98 and TA100 strains. These findings add evidence that chañar fruit flour may be considered a functional food with preventive properties against diseases associated with oxidative stress, inflammatory mediators and metabolic syndrome.

Design and quality control of a pharmaceutical formulation containing natural products with antibacterial, antifungal and antioxidant properties

The aims of the present study were to determine the antibacterial and antifungal activity as well as mutagenicity of Sechium edule fluid extract and to obtain a pharmaceutical formulation with them. The extract exhibited antimicrobial activity against Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Enterobacter cloacae, Serratia marcescens, Morganella morganii, Acinetobacter baumannii, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Candida spp. and Aspergillus spp. isolated from clinical samples from two hospitals of Tucuman, Argentina. Non-toxicity and mutagenicity on both Salmonella typhimurium TA98 and TA 100 strains until 100 microg/plate were observed. A hydrogel with carbopol acrylic acid polymer containing S. edule fluid extract as antibacterial, antimycotic and antioxidant agent was obtained. Microbiological, physical and functional stability of pharmaceutical formulation conserved at room temperature for 1 year were determined. Addition of antioxidant preservatives to store the pharmaceutical formulation was not necessary. The semisolid system showed antimicrobial activity against all gram positive and gram negative bacteria and fungi assayed. The minimal inhibitory concentration (MIC) values ranged from 20 to 800 microg/mL. Its activity was compared with a pharmaceutical formulation containing commercial antibiotic and antifungal. A pseudoplastic behavior and positive thixotropy were observed. Our current finding shows an antimicrobial activity of hydrogel containing S. edule extract on a large range of gram negative and gram positive multi-resistant bacteria and fungi. This topical formulation may be used as antimycotic and as antibacterial in cutaneous infections.

ANTIMICROBIAL AND ANTIOXIDANT COMPOUNDS FROM THE INFUSION AND METHANOLIC EXTRACT OF Baccharis incarum (WEDD.) PERKINS.

In the traditional medicine of the South American highlands, the infusion and decoction of aerial parts of “tola”, Baccharis incarum (Wedd.) Perkins (Asteraceae) is orally taken as an antiseptic, antipyretic and digestive or externally applied to relieve pain and inflammation. A plant infusion and methanolic (MeOH) extract of the aerial parts were compared for main constituents by HPLC-MS as well as assessed for antimicrobial and free radical scavenging activities. Assay-guided fractionation of the polar extracts was carried out using the ABTS•+ autographic as well as methicillin-resistant Staphylococcus aureus and Enterococcus faecalis bioautographic test. Seven compounds were isolated from the polar extracts and identified as chlorogenic acid, 3’,4’,5,7-tetrahydroxyflavone; dicaffeoyl quinic acid; 3’,4’,5,7-tetrahydroxy-3,6-dimethoxyflavone; 3’,4’,5,7-tetrahydroxy-3,6,8-trimethoxy flavone, 4’,5,7trihydroxy-3’,3,6,8-tetramethoxyflavone and 4’,5-dihydroxy-3’,3,6,7,8-pentamethoxyflavone . The main constituents from the methanol extract and infusion were caffeic acid derivatives. This is the first report on the constituents of B. incarum infusion and show clear differences with previous phytochemical studies on the same plant. All isolated compounds showed antioxidant activity with SC 50 values of 1 to 10 µg/ml. The isolated flavones were active against methicillin-resistant Staphylococcus aureus and Enterococcus faecalis with MICs in the range 100 and >200 µg/ml.

Flour from Prosopis alba cotyledons: A natural source of nutrient and bioactive phytochemicals

The Prosopis alba seed is a waste material in the process to produce pod flour. To suggest a potential use of these seeds it is necessary to determine the nutritional, phytochemical and functional quality of cotyledon flour from Prosopis alba. This flour showed high level of proteins (62%), low content of total carbohydrate and fat. Free polyphenol (1150±20mg GAE/100g flour) and carotenoids (10.55±0.05mg β-CE/100g flour) compounds were the dominant compounds. The main identified constituents in the polyphenolic extracts were C- glycosyl flavones, including schaftoside, isoschaftoside, vicenin II, vitexin and isovitexin. The extract enriched in polyphenolic compounds exhibited ABTS(+) reducing capacity and scavenging activity of H2O2; and was able to inhibit phospholipase, lipoxygenase and cyclooxygenase, three pro-inflammatory enzymes. According to our results, the P. alba cotyledon flour could be considered as a new alternative in the formulation of functional foods or food supplements.

Polyphenolic compounds and anthocyanin content of Prosopis nigra and Prosopis alba pods flour and their antioxidant and anti-inflammatory capacities

The aim of this study was to determine the content of total free and bound phenolics, free and bound flavonoids, anthocyanins, and alkaloids and the profile of polyphenols of edible ripe pods of Prosopis alba and Prosopis nigra. P. alba flour showed significantly higher total (sum of Free- and Bound) phenolic content and total flavonoid compounds than P. nigra (p<0.05) while P. nigra had higher concentrations of anthocyanins than P. alba (p<0.05). The P. nigra flour shows a pattern characterized by the occurrence of anthocyanins as well as 14 flavonoid glycosides, with higher chemical diversity than P. alba, which shows 8 flavonoid glycosides as relevant constituents. The main compounds were quercetin O-glycosides and apigenin-based C-glycosides. The phenolic composition of two South American algarrobo pod flour is presented for the first time. P. nigra pods having higher content of anthocyanins are darker (purple) than those of P. alba (light brown). Furthermore, the sugar-free polyphenolic extracts of P. nigra and P. alba as well as anthocyanins enriched extracts from P. nigra showed antioxidant activity. P. nigra and P. alba polyphenolic extracts showed activity against a pro-inflammatory enzyme. In conclusion, algarrobo pods meal contained biologically active polyphenols, with a positive impact on human health.

Chemical and functional characterization of seed, pulp and skin powder from chilto (Solanum betaceum), an Argentine native fruit. Phenolic fractions affect key enzymes involved in metabolic syndrome and oxidative stress

The aim of this work was to assess the nutritional and functional components of powder obtained by lyophilization of whole fruits, seeds, pulp and skin from chilto (Solanum betaceum Cav) cultivated in the ecoregion of Yungas, Argentina. The powders have low carbohydrate and sodium content and are a source of vitamin C, carotenoid, phenolics, potassium and fiber. The HPLC-ESI-MS/MS analysis of the fractions enriched in phenolics allowed the identification of 12 caffeic acid derivatives and related phenolics, 10 rosmarinic acid derivatives and 7 flavonoids. The polyphenols enriched extracts before and after simulated gastroduodenal digestion inhibited enzymes associated with metabolic syndrome, including α-glucosidase, amylase and lipase and exhibited antioxidant activity by different mechanisms. None of the analyzed fruit powders showed acute toxicity or genotoxicity. The powders from the three parts of S. betaceum fruit may be a potential functional food and the polyphenol enriched extract of seed and skin may have nutraceutical properties.

Antioxidant/Antibacterial Activities of a Topical Phytopharmaceutical Formulation Containing a Standardized Extract of Baccharis incarum, an Extremophile Plant Species from Argentine Puna

The antimicrobial and antioxidant activities of standardized extracts of Baccharis incarum in 60° and 80° ethanol and of a phytopharmaceutical formulation obtained from them were measured. Baccharis tinctures showed antimicrobial activity against clinically isolated antibiotic resistant Staphylococcus aureus and Enterococcus faecalis, with MIC values of 40–80 µg GAE/mL. Both tinctures exhibited ABTS●+ scavenging activity with SC50 values between 1.6 and 4.0 µg GAE/mL. The tinctures were not genotoxic in the Salmonella assay. For this reason, the tincture in 60° ethanol was incorporated into a topical pharmaceutical formulation (Hydrogel/ Carbopol® 934). The phytopharmaceutical formulation also showed antibacterial and antioxidant activities in the in vitro assays. The hydrogel showed microbiological, chemical, physical and functional stability during storage at room temperature. Studies that measure drug release as a determination of bioavailability were also carried out using the Franz diffusion cell (FC). The results demonstrated the release of two bioactive compounds (chlorogenic acid and 4′,5‐dihydroxy‐3′,3,6,7,8‐pentamethoxyflavone) from the phytotherapic preparation in HPLC studies of FC receptor solution. In consequence, the phytopreparation applied topically could be used to treat skin and soft tissue infection produced by methicillin‐resistant Staphylococcus aureus (MRSA) or Enterococcus faecalis and opens new opportunities for the use of active natural ingredients in the cosmeceutical field as antiacne and antioxidant. Copyright

Analytical methodology optimization to estimate the content of non-flavonoid phenolic compounds in Argentine propolis extracts

Abstract Context: Traditionally, the content of total phenolics (flavonoid phenolics (FP) and non-flavonoid phenolics (NFP)) and flavonoids (flavone/flavonol and flavonone/dihydroflavonol) in propolis has been determined by different methodologies. Until now, the percentage of total phenolic (TP) compounds that corresponds to FP and NFP, expressed in the same units by a spectrophotometric method, has not been determined. Objective: The current study proposes a quick and simple methodology that separates FP and NFP in propolis samples and determines TP, FP, and NFP by the same method. Materials and methods: Propolis samples from five Argentine provinces (Tucumán, Santiago del Estero, Salta, Misiones, and Jujuy) were used. Extraction of TP from the propolis samples was carried out by maceration with 80% ethanol and quantified by Folin–Ciocalteu reagent (FC-R). Then, FP was precipitated with formaldehyde in acid medium. After centrifugation, NFP were determined in the supernatant using FC-R. FP content was calculated as the difference between the content of TP and NFP. The method was also validated using commercial flavonoids and chalcones. Results: FP recovery in all experiments was between 85.95% and 98.29%. Propolis from Tucumán had significantly higher amounts of total phenols than propolis from other provinces. SE5 showed higher content of FP (81.52%) followed by SA1 (74.75%). The propolis from TUC4, SA4, SE3, and MI showed the lowest FP content and highest content of NFP. Conclusions: This method provides a simple, reliable, and specific spectrophotometric assay to estimate the content of NFP, FP, and TP in propolis samples.

In vitro antimicrobial activity of 20 selected climber species from the Bignoniaceae family.

Hydroalcoholic and aqueous extracts of some climber species from the Bignoniaceae family that grow in the north of Argentina were evaluated for in vitro antibacterial activity against Gram-positive and Gram-negative strains. By means of bioautography and disc diffusion methods, it could be determined that all infusions were not active, whereas the hydroalcoholic extracts of seven species were able to inhibit bacterial growth. The minimum inhibitory concentration and minimum bactericidal concentration observed were between 62.5 and 1000 μg gallic acid equivalent (GAE)/mL and between 125 and 1000 μg GAE/mL, respectively. The tested extracts were more active against Gram-positive microorganisms. Time-kill experiments indicated that all extracts have bacteriostatic activity. Phytochemical screening showed the presence of terpenoids, phenols and flavonoids. The amount of phenolic compounds and flavonoids was higher in tinctures when compared with infusions. These results suggest the presence of antibacterial substances in the hydroalcoholic extracts, which could be used for the treatment of infections.