The aim of this study was to assess, using the DPPH assay, the antioxidant activity of several substances that could be proposed to immediately revert the problems caused by bleaching procedures. The percentage of antioxidant activity (AA%) of 10% ascorbic acid solution (AAcidS), 10% ascorbic acid gel (AAcidG), 10% sodium ascorbate solution (SodAsS), 10% sodium ascorbate gel (SodAsG), 10% sodium bicarbonate (Bicarb), Neutralize(®) (NE), Desensibilize(®) (DES), catalase C-40 at 10 mg/mL (CAT), 10% alcohol solution of alpha-tocopherol (VitE), Listerine(®) (LIS), 0.12% chlorhexidine (CHX), Croton Lechleri (CL), 10 % aqueous solution of Uncaria Tomentosa (UT), artificial saliva (ArtS) and 0.05% sodium fluoride (NaF) was assessed in triplicate by 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) free radical assay. All substances exhibited antioxidant activity, except for CL. AAcidS, AAcidG and VitE exhibited the highest AA% (p<0.05). On the contrary, CHX, NE, LIS and NaF showed the lowest AA% (p<0.05). In conclusion, AAcidS, AAcidG, SodAsS, SodAsG and VitE presented the highest antioxidant activity among substances tested in this study. The DPPH assay provides an easy and rapid way to evaluate potential antioxidants.
Isolation and identification of compounds with antioxidant activity in a novel variety of Brazilian propolis produced by bees from the species Apis mellifera Propolis is known for its wide range of biological activities, such as antioxidant, antibacterial, anti-inflammatory, antiviral, among others. Its composition varies according to the region where it is collected and there is a great chemical diversity among the different types of propolis. The main constituents of propolis are the phenolic compounds, which are responsible for several biological activities, such as the antioxidant activity. A novel type of propolis, which has never been reported in the literature before, named red propolis due to its intense characteristic color, was collected in a mangrove area in the state of Alagoas. This study aimed to evaluate the chemical composition, and fractionate and isolate the compounds with antioxidant activity of Brazilian red propolis. The Ethanolic Extract of Propolis (EEP) was first fractionated using the liquid-liquid extraction technique, yielding the hexanic (Hex-fr) and the chloroformic fractions (Chlo-fr). Chlo-fr showed great yield and a complex chemical composition and therefore was refractioned in silica gel open column, yielding 15 subfractions. The EEP, fractions and subfractions were evaluated for their chemical composition and antioxidant activity aiming to select the fractions destined for bioguided isolation. The EEP was also analyzed by the gas-chromatography-mass spectrometry (GC-MS) technique, and seven compounds, never reported for Brazilian propolis before, were identified, among which are four isoflavonoids. The amounts of total polyphenol and flavonoid compounds, tested for EEP, fractions and subfractions, ranged from 37,4 to 324,2 mg/g and 0,0 to 158 mg/g, respectively. The major free radical (DPPH) scavenging activity was found for Hex-fr, while subfraction 5 showed high yield and good antioxidant activity by β-carotene/linoleic acid system oxidation, and these fractions were selected for isolation of bioactive compounds by preparative High Performance Liquid Chromatography (HPLC). Three compounds, one from Hex-fr and two from subfraction 5, were isolated and analyzed by GC-MS. The compounds from subfraction 5 were identified as 2 ' ,7-diidroxi-4 ' metoxiisoflavan (vestitol) and 2 ' ,4 '-dihidroxi-7 metoxiisoflavan, both isoflavonoids. It was not possible to identify the compound from Hex-fr using GC-MS, although it was possible to verify that this compound is a mixture of two isomers presenting molecular weight of 466 g/mol. The results obtained in this study demonstrate that red propolis is a good source of bioactive compounds, among which are isoflavonoids, compounds that possess antioxidant activity.
The antibiotic susceptibility of members of the family Enterobacteriaceae and of Staphylococcus aureus strains isolated from the respiratory tract was assessed over the period 1997-1999 as part of the Italian Epidemiological Observatory survey sponsored by the Smith-Kline Foundation. A standardised method was used to determine the MICs of 22 antibiotics against isolates of Klebsiella pneumoniae (n=870), Escherichia coli (n=684), Enterobacter cloacae (n=342), Enterobacter aerogenes (n=187) and Serratia marcescens (n=135) as well as the MICs of 11 antibiotics against isolates of Staphylococcus aureus (n=1,606). Overall, the susceptibility rate of Enterobacteriaceae isolates was > or = 90% to 5 agents (meropenem, imipenem, amikacin, cefepime and gentamicin); 89-80% to 2 agents (ciprofloxacin and tobramycin); and <80% to 11 agents (cefotaxime, piperacillin-tazobactam, trimethoprim-sulfamethoxazole, cefetamet, ceftriaxone, ceftazidime, aztreonam, ticarcillin-clavulanate, tetracycline, piperacillin, cefuroxime, chloramphenicol, ticarcillin, amoxicillin-clavulanate and amoxicillin). During the 3-year monitoring period, antibiotic susceptibility increased in Klebsiella pneumoniae against amoxicillin-clavulanate, in Escherichia coli against third-generation cephalosporins and aztreonam, in Enterobacter aerogenes against amoxicillin and piperacillin-tazobactam and in Serratia marcescens against most of the antibiotics. In contrast, Enterobacter cloacae showed a tendency to develop resistance to cefetamet, amikacin and ciprofloxacin. Of the total number of Staphylococcus aureus strains, 38% were methicillin resistant. Nearly 80% of the methicillin-resistant strains displayed a multiresistance pattern (additional resistance to 2 or more non-beta-lactam antibiotics). Rates of susceptibility of particular species (Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus) were compared using strains from different geographical areas of Italy (northern, central and southern) and from different nosocomial areas (outpatients, intensive care unit [ICU] inpatients, non-ICU inpatients). Susceptibility of Klebsiella pneumoniae to several antibiotics was lower in southern Italy, whereas the incidence of methicillin-resistant strains was higher in northern and central Italy. The susceptibility of Escherichia coli was similar in all three areas. No significant differences in susceptibility of Klebsiella pneumoniae or Escherichia coli were found between strains from inpatients and outpatients or from inpatients admitted to ICU and non-ICU units. The incidence of methicillin-resistant Staphylococcus aureus was higher in ICU inpatients (52%) than in non-ICU inpatients (38%) and lower in outpatients (19%) than in inpatients.
Este trabalho teve como objetivo avaliar o teor de compostos fenólicos totais e a atividade antioxidante de chás consumidos no Brasil. Avaliou-se o teor de compostos fenólicos totais pelo método do Folin Ciocalteu e a atividade antioxidante pelo sequestro do radical DPPH. (porcentagem de inibição do radical DPPH.; porcentagem de DPPH. remanescente; coeficiente de inibição, IC50). As análises das seguintes infusões foram realizadas em triplicata: chá preto, chá verde, mate, camomila e anis (erva-doce) de três diferentes marcas (A, B e C). Os maiores teores de compostos fenólicos foram encontrados em chá verde (C e B), chá preto (C) e mate (A) (teores superiores a 65 mg EAG / g de chá). O chá verde (C e B) foi o mais eficiente em termos de porcentagem de inibição do radical DPPH. (valores acima de 90%), de porcentagem de DPPH. remanescente (menos de 10% da concentração inicial) e em termos de IC50 (valores inferiores a 150 µg de massa seca/mL de infusão). O chá de camomila, percentualmente, apresentou os melhores índices, mas o parâmetro IC50 resultou em valores elevados que correspondem à baixa atividade antioxidante. Não houve correlação entre os teores de compostos fenólicos e a atividade antioxidante demonstrada pela percentagem de inibição e pela percentagem de DPPH. remanescente. No entanto a correlação entre os teores de compostos fenólicos e a atividade antioxidante evidenciada pelo coeficiente de inibição (IC50) foi elevada (r=0,92), indicando a existência de compostos com distintos potenciais antioxidantes.
The aim of this study was to evaluate the most effective extraction condition (temperature, solvent type and time) for recovery of high-value phytochemicals present in the Tabernaemontana catharinensis leaves (TC) and to assess their effect on biochemical parameters in streptozotocin-induced diabetic rats. The extraction of phenolic compounds from TC using a factorial design (FD) 2³, high performance liquid chromatography (HPLC), response surface methodology (RSM) and principal component analysis (PCA) were studied. It was found that the optimal conditions for extraction of phenolics were higher temperature (65 °C) and time (60 min) using ethanol as extractor solvent. In this condition of extraction (A8), total phenolic compounds (TPC) and antioxidant activity (AA) were determined. Additionally, this extract was used to evaluate their effect on antioxidant enzyme activities (superoxide dismutase (SOD) and catalase (CAT)) as well as lipid peroxidation (LP) and protein thiols level (PSH) in the liver and kidneys of normal and diabetic rats. As result, T. catharinensis extract presented TPC content of 23.34 mg EAG/g (equivalent gallic acid) and AA of 34.26 μmol Trolox/g. Phenolic acids (ferulic acid and coumaric acid) and flavonoids (quercetin, rutin and pinocembrin) could be recovered and identified by HPLC. This study indicated an important role of the T. catharinensis extract on free radical inactivation and on the antioxidant defense system in diabetic rats. In fact, the use of T. catharinensis extract restored the normal activity of SOD (p < 0.05) and suppressed malondialdehyde levels in liver and kidney tissues. Thus, the T. catharinensis extract, rich in phenolic compounds, can be responsible for the recover the enzymatic changes in the liver and kidney tissues provoked by diabetes in rats. In addition, the lipid peroxidation rate decreased in the diabetic rats treated with T. catharinensis.
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