Evaluation of antiradical assays used in determining the antioxidant capacity of pure compounds and plant extracts

The efficiency of the chemiluminescence luminol method and colorimetric DPPH and ABTS methods in evaluating the antiradical capacity of pure compounds and plant extracts with antioxidant potential is compared. In case of pure compounds, the values of parameter 'n' (number of radicals quenched per molecule of antiradical) for ascorbic acid, p-hydroquinone, catechol, quercetin, and rutin are similar when measured by colorimetric assays; however, considerably lower values of n are obtained with the luminol assay. The antiradical activity of extracts from male and female individuals of Baccharis burchelli and Baccharis crispa were determined by the luminol assay and expressed using the new Trolox® percentage (%Trolox®) parameter.

Desenvolvimento de um sistema de análise por injeção em fluxo utilizando materiais alternativos de baixo custo para fins didáticos

This work describes a simple and inexpensive flow injection analysis system in which gravity force provides fluid propulsion while needles for insulin administration or metallic wires act as electrodes for amperometric detection. The proposed system was able to demonstrate the influence of several operational parameters on the transient signals. Moreover, this system was successfully used to evaluate both the stoichiometry of Cu2+-EDTA complex and the effect of pH on the kinetics of the reaction between ferricyanide and ascorbic acid. Therefore, the proposed system can be regarded as an efficient and accessible didactic tool for the teaching of FIA principles.

Interação de complexo de rutênio com albumina de soro bovino para detecção amperométrica de ácido ascórbico

The binding of [Ru(PAN)(PPh3)2(ISN)]Cl (PAN = 1-(2'-Pyridylazo)-2-naphtholate) to bovine serum albumin (BSA) was investigated by spectroscopic techniques. According to analysis of the results from the Stern-Volmer equation, the ruthenium complex is able to quench the fluorescence intensity of BSA via a dynamic mechanism. The thermodynamic parameters were calculated (ΔH = 30.3 kJ mol-1; ΔS = 195.4 J mol-1 K-1), indicating that hydrophobic force is the main interaction driving force. The site marker competitive experiments revealed that the binding site of ruthenium complex was in the sub-domain IIA of BSA. FTO glass with a film of BSA-[Ru(PAN)(PPh3)2(ISN)]Cl was used as an ascorbic acid sensor. The linear range of the modified electrode was between 1 and 8 × 10-6 mol L-1.

ENCAPSULAÇÃO DO ÁCIDO L-ASCÓRBICO NO BIOPOLÍMERO NATURAL GALACTOMANANA POR SPRAY‑DRYING: PREPARAÇÃO, CARACTERIZAÇÃO E ATIVIDADE ANTIOXIDANTE

AbstractIn this study, the spray drying technique was used to prepare L-ascorbic acid (AA) microparticles encapsulated with galactomannan-an extract from the seeds of the Delonix regia species. The physico-chemical characteristics, antioxidant activity, and encapsulation efficiency of the AA microparticles were evaluated and characterized using thermogravimetric analysis, differential scanning calorimetry, infrared spectroscopy, X-ray diffraction, and scanning electron microscopy. The free-radical scavenging activity of the AA microparticles was determined at different environmental conditions using DPPH (1,1-diphenyl-2-picryl-hydrazyl). X-ray diffraction measurements demonstrated a loss of crystallinity in AA after the encapsulation process, and a DSC scan also showed the loss of the compound's melting peak. Thermogravimetric analysis showed small differences in the thermal stability of galactomannan before and after the incorporation of AA. The mean diameters of the obtained spherical microspheres were in the range of 1.39 ± 0.77 µm. The encapsulation efficiency of AA microparticles in different environmental conditions varied from 95.40 to 97.92, and the antioxidant activity showed values ranging from 0.487 to 0.550 mg mL-1.

Study of complexes of cadmium with some L-amino acids and vitamin-C by voltammetric technique

Voltammetric technique was used to study the binary and ternary complexes of cadmium with L-amino acids and vitamin-C (L-ascorbic acid) at pH =7.30 ± 0.01, µ = 1.0M KNO3 at 25ºC and 35ºC. Cd (II) formed 1:1:1, 1:1:2 and 1:2:1 complexes with L-lysine, L-ornithine, L-threonine, L-serine, L-phenylglycine, L-phenylalanine, L-glutamic acid and L-aspartic acid used as primary ligands and L-ascorbic acid used as secondary ligand. The trend of stability constant of complexes was L-lysine < L-ornithine < L-threonine < L-serine < L-phenylglycine < L-phenylalanine < L-glutamic acid < L-aspartic acid which can be explained on the basis of size, basicity and steric hindrance of ligands. The values of stability constant (log β) varied from 2.23 to11.33 confirm that these drugs i.e. L-amino acids or in combination with L-ascorbic acid or their complexes could be used against Cd (II) toxicity. The study has been carried out at 35ºC also to determine the thermodynamic parameters such as enthalpy change (ΔH), Free energy change (ΔG) and entropy change (ΔS) respectively.

Chitosan and fungicides on postharvest control of Guignardia citricarpa and on quality of 'Pêra Rio' oranges

Citrus fruits are affected by the black spot disease caused by the fungus Guignardia citricarpa. Chitosan can be used as covering for fruits and may delay the ripening process and inhibit the growth of some fungi. Thus, the control of citrus black spot using chitosan and the fungicides thiabendazole and imazalil was assessed in addition to the physicochemical quality of 'Pêra Rio' oranges. The oranges were immersed into chitosan, thiabendazole or imazalil, and in chitosan mixed with both fungicides. The fruits were then stored at 25 °C, 80% RH, for 7 days and, after this storage period, subjected to physicochemical analyses. Chitosan in association with the fungicides reduced black spot in 'Pêra Rio' oranges and delayed the change in the orange skin colour from green to yellow during the postharvest storage. Total soluble solids, titratable acidity, pH, ascorbic acid content and ratio were not influenced by the treatments. Thus, chitosan applied with the fungicides thiabendazole and imazalil showed potential to control the development of black spot lesions on 'Pêra Rio' oranges during the postharvest period.

Microencapsulation of pequi pulp by spray drying: use of modified starches as encapsulating agent

The aim of this study was to evaluate the microencapsulation of pequi pulp by spray drying. A central composite rotational design was used in order to evaluate the effect of the independent variables: inlet air temperature, surfactant concentration and modified starch concentration. The dependent variables were assumed as yield of the process and the product features microencapsulated. A selection of the best process condition was performed to obtain the best condition of a product with the highest vitamin C and carotenoids content. Powders showed moisture content below 2%. The experimental values of hygroscopicity, yield, water activity, total carotenoids and vitamin C powders ranged from 7.96 to 10.67 g of adsorbed water/100g of solids, 24.34 to 49.80%, 0.13 to 0.30, 145.78 to 292.11 mg of ascorbic acid/g of pequi solids and 15.51 to 123.42 mg of carotenoids/g of pequi solids, respectively. The inlet air temperature 140°C, the surfactant concentration of 2.5% and the modified starch concentration of 22.5% was recommended as the selected condition. By the scanning electron microscopy, it was observed that most of the particles had spherical shape and smooth surface.

Mechanical damage during harvest and loading affect orange postharvest quality

Brazil is the world’s largest orange producer; however, part of this production is lost during postharvest. This loss can be minimized by controlling incidence of physical damage throughout the harvest and loading operations. Impacts can negatively modify quantitative and qualitative fruits aspects. The main goal of this study was to measure the impact magnitude in two types of harvest (manual and detachment) and during all steps from picking into bags until loading for transport to the processing industry and additionally evaluating, in laboratory, the physico-chemical quality of the fruit subjected to various impacts, similar to those found in the field. In order to evaluate the impact magnitude, an instrumented sphere was used (760 mm, Techmark, Inc, USA). The following physico-chemical parameters were evaluated during 6-days of storage: weight loss, soluble solids contents, titratable acidity, ascorbic acid content, pH, firmness and peel color. The greatest impacts were observed during harvest, during the detachment practice, and when loading and unloading from bulk storage, with average acceleration values between 249.5 and 531.52G. The impact incidence in oranges were responsible for reducing the soluble solids, titratable acidity, ascorbic acid and weight by to 5.5%; 8.7%; 4.6% and 0.5%, respectively, compared to the control. Impacts during harvest and the various pre-industry manipulation steps must be controlled as they interfere in postharvest quality and physiology of ‘Valência’ oranges.

Jalometallien pelkistyskemikaalit

Tässä työssä esitellään kirjallisuudesta löytyneitä vaihtoehtoja jalometallien (Au, Ag, Pt, Pd, Is, Os, Rh, Ru, Re) homogeeniseen pelkistämiseen ja tekijöitä, jotka vaikuttavat pelkistimen valintaan. Jalometallien korkea hinta tekee niiden talteenoton pienistäkin pitoisuuksista kiinnostavaksi. Pelkistäminen on talteenoton viimeinen vaihe, jota voidaan käyttää myös puhdistusaskeleena. Pelkistimen valinnalla on suuri merkitys pelkistystulokseen. Myös pH:lla ja lämpötilalla on merkittävä vaikutus pelkistykseen. Ideaalinen pelkistyskemikaali on edullinen, selektiivinen, sillä on kohtuullinen pelkistysaika, se ei ole haitallinen, ei muodosta haitallisia sivutuotteita ja tarvittava prosessi on yksinkertainen. Lupaavia pelkistyskemikaaleja ovat esimerkiksi askorbiinihappo, vetyperoksidi ja muurahaishappo.

Kullan talteenotto takaisinuuttoliuoksista

Työssä tutkittiin kirjallisuuden ja laboratoriomittausten avulla vaihtoehtoja kullan pelkistämiseen ja talteenottoon kultauuton takaisinuuttoliuoksista. Tavoitteena oli löytää menetelmä, jolla saadaan puhdasta kiinteää lopputuotetta ilman kullan häviöitä. Käytettyjä pelkistimiä olivat D-(+)-glukoosi, natriumboorihydridi, L-askorbiinihappo, D-(-)-isoaskorbiinihappo ja aktiivihiili. Laboratoriokokeiden perusteella D-(-)-isoaskorbiinihappo sekä aktiivihiili olivat sopivimmat pelkistimet kokeissa käytetylle kultaliuokselle. Isoaskorbiinihapolla suoritettiin panoskokeita lasireaktorissa eri alku-pH:ssa sekä erilaisilla pelkistimen ja kullan moolisuhteilla. Tulosten perusteella havaittiin pH:n ja pelkistimen ylimäärän vaikuttavan merkittävästi lopputuotteen puhtauteen. Myös redox-potentiaalia säätämällä ja happopesulla pelkistyksen jälkeen voidaan vaikuttaa lopputuotteen puhtauteen. Aktiivihiilellä suoritettiin panoskokeita adsorptiotasapainojen (latausisotermi) ja kinetiikan tutkimiseksi. Hiileen on mahdollista saada kultaa 383 mg/g kuivaa hiiltä. Suurempi lataus voitaisiin saavuttaa käyttämällä hiiltä, jolla on pienempi partikkelikoko. Kolonnikokeita tehtiin eri virtausnopeuksilla. Kolonnikokeissa kullan dynaaminen adsorptiokapasiteetti hiileen odotetusti kasvoi virtausnopeuden laskiessa. Pienin käytetty virtausnopeus oli 2,40 BV/h, jolloin kapasiteetti oli 75,4 mg/g kuivaa hiiltä (c (Au feed) = 129 mg/L). Kullasta voidaan poistaa myös kolonnipelkistyksen jälkeen epäpuhtauksia happopesulla. Isoaskorbiinihapolla pelkistyksen kinetiikka on nopea ja sillä saatiin pelkistettyä puhdasta lopputuotetta. Sekä isoaskorbiinihappo, että aktiivihiili ovat potentiaalisia menetelmiä kullan talteenottoon.

Antioxidant status of dog aqueous humor after extracapsular lens extraction

We determined the antioxidant status of the aqueous humor after extracapsular lens extraction in 14 mongrel dogs weighing about 10 kg. The animals were examined by slit lamp biomicroscopy, applanation tonometry and indirect ophthalmoscopy. One eye was submitted to conventional extracapsular lens extraction and the other was used as control. Samples of aqueous humor were obtained by anterior chamber paracentesis before and at days 1, 2, 3, 7 and 15 after surgery. Total antioxidant status was determined as the capacity of aqueous humor to inhibit free radical generation by 2,2-azobis(2-amidopropane) chlorine. Ascorbic acid concentration was measured by HPLC with UV detection. Protein content was determined with the biuret reagent. Statistical analysis was performed by ANOVA followed by the Tukey-Kramer test. Protein concentration increased from 0.61 to 22 mg/ml 24 h after surgery. These levels were maintained and returned to normal at day 7. Total antioxidant capacity was reduced from 50 to about 30 min until day 3 and at day 7 it was equal to control. Ascorbic acid levels were reduced from 252 to about 110 µM and then returned to control values at day 15. Considering the importance of ascorbic acid concentration in aqueous humor for the maintenance of the antioxidant status of the anterior segment of the eye, the decrease of antioxidant defenses suggests that the surgical procedures promote an oxidative stress condition in the eye.

Selegiline increases heme oxygenase-1 expression and the cytotoxicity produced by dopamine treatment of neuroblastoma SK-N-SH cells

Increased dopamine catabolism may be associated with oxidative stress and neuronal cell death in Parkinson's disease. The present study was carried out to examine the effect of dopamine on the expression of heme oxygenase-1 and -2 (HO-1 and HO-2) in human neuroblastomas (SK-N-SH cell line) and the effects of selegiline and antioxidants on this expression. Cells were kept with close control of pH and were incubated with varying concentrations of dopamine (0.1-100 µM) for 24 h. HO-1 and HO-2 cDNA probes were prepared by reverse transcription-polymerase chain reaction amplification. The mRNA expression of HO-1 and HO-2 was measured by Northern blot analysis. The levels of HO-1 mRNA increased after dopamine treatment, in a dose-dependent manner, in all cell lines studied, whereas levels of the two HO-2 transcripts did not. The HO-1 and HO-2 protein expression was analyzed by Western blotting. HO-1 protein was undetectable in untreated SK-N-SH cells and increased after treatment with dopamine. In contrast, the HO-2 protein (36 kDa) was detected in untreated cells and the levels did not change as a result of treatment. alpha-Tocopherol (10-100 µM) and ascorbic acid (100 µM) did not attenuate the effects of dopamine. Selegiline (10 µM) produced significant increase (P < 0.01) in the induction of HO-1 by dopamine (more than six times the control values). The increased expression of HO-1 following dopamine treatment indicates that dopamine produces oxidative stress in this cell line.

Electron paramagnetic resonance study of lipid and protein membrane components of erythrocytes oxidized with hydrogen peroxide

Electron paramagnetic resonance (EPR) spectroscopy of spin labels was used to monitor membrane dynamic changes in erythrocytes subjected to oxidative stress with hydrogen peroxide (H2O2). The lipid spin label, 5-doxyl stearic acid, responded to dramatic reductions in membrane fluidity, which was correlated with increases in the protein content of the membrane. Membrane rigidity, associated with the binding of hemoglobin (Hb) to the erythrocyte membrane, was also indicated by a spin-labeled maleimide, 5-MSL, covalently bound to the sulfhydryl groups of membrane proteins. At 2% hematocrit, these alterations in membrane occurred at very low concentrations of H2O2 (50 µM) after only 5 min of incubation at 37°C in azide phosphate buffer, pH 7.4. Lipid peroxidation, suggested by oxidative hemolysis and malondialdehyde formation, started at 300 µM H2O2 (for incubation of 3 h), which is a concentration about six times higher than those detected with the probes. Ascorbic acid and α-tocopherol protected the membrane against lipoperoxidation, but did not prevent the binding of proteins to the erythrocyte membrane. Moreover, the antioxidant (+)-catechin, which also failed to prevent the cross-linking of cytoskeletal proteins with Hb, was very effective in protecting erythrocyte ghosts from lipid peroxidation induced by the Fenton reaction. This study also showed that EPR spectroscopy can be useful to assess the molecular dynamics of red blood cell membranes in both the lipid and protein domains and examine oxidation processes in a system that is so vulnerable to oxidation.

2-Bromo-1,4-naphthoquinone: a potentially improved substitute of menadione in Apatone™ therapy

Apatone™, a combination of menadione (2-methyl-1,4-naphthoquinone, VK3) and ascorbic acid (vitamin C, VC) is a new strategy for cancer treatment. Part of its effect on tumor cells is related to the cellular pro-oxidative imbalance provoked by the generation of hydrogen peroxide (H2O2) through naphthoquinone redox cycling. In this study, we attempted to find new naphthoquinone derivatives that would increase the efficiency of H2O2 production, thereby potentially increasing its efficacy for cancer treatment. The presence of an electron-withdrawing group in the naphthoquinone moiety had a direct effect on the efficiency of H2O2 production. The compound 2-bromo-1,4-naphthoquinone (BrQ), in which the bromine atom substituted the methyl group in VK3, was approximately 10- and 19-fold more efficient than VK3 in terms of oxygen consumption and H2O2 production, respectively. The ratio [H2O2]produced / [naphthoquinone]consumed was 68 ± 11 and 5.8 ± 0.2 (µM/µM) for BrQ and VK3, respectively, indicating a higher efficacy of BrQ as a catalyst for the autoxidation of ascorbic acid. Both VK3 and BrQ reacted with glutathione (GSH), but BrQ was the more effective substrate. Part of GSH was incorporated into the naphthoquinone, producing a nucleophilic substitution product (Q-SG). The depletion of BrQ by GSH did not prevent its redox capacity since Q-SG was also able to catalyze the production of reactive oxygen species. VK3/VC has already been submitted to clinical trials for the treatment of prostate cancer and has demonstrated promising results. However, replacement of VK3 with BrQ will open new lines of investigation regarding this approach to cancer treatment.

Postharvest respiratory activity and changes in some chemical constituents during maturation of yellow mombin (Spondias mombin) fruit

Mature fruit from the yellow mombin (Spondias mombin) was monitored for its respiration activity. Mature green fruit from the yellow mombin was stored in closed glass chambers and the concentration of oxygen and carbon dioxide at the end of a six hour respiration period was determined. At the same interval of time, the lid of the chamber was opened for air renewal. The increase in carbon dioxide and decrease in oxygen concentration demonstrated that the fruit was climacteric. The maximum liberation of CO2 54.2 mL Kg-1 h-1 and maximum absorption of O2 49.0 mL Kg-1 h-1 occurred 186 hours after the harvest which, obviously, represented the optimum fruit quality after the senescence process started. The respiratory quotient of fruit at a climacteric maximum was 1.11 representing the oxidation of carbohydrates. Total soluble solids increased from 9.1 °Brix (initial) to 13.7 °Brix (climacteric maximum) during maturation, while the total number of acids in the fruit decreased during maturation i.e. from 1.55% initially to 1.40% at pre-climacteric, 1.0% at climacteric maximum and 0.8% in the post-climacteric stage. A similar behaviour was observed in the case of ascorbic acid. There was a continuous decrease in chlorophyll and a continuous increase in the carotenoid content of fruit during maturation.