Angela Fadda

Determination of Free Radical Scavenging Activity of Plant Extracts Through DPPH Assay: An EPR and UV–Vis Study

Several methods have been developed to assess the radical scavenging activity. Among them, the 2,2-diphenyl-1-picrylhydrazyl (DPPH) spectrophotometric method is one of the most widely applied and is appreciated for its reliability. In this study, a comparison of two spectroscopic methods (electron paramagnetic resonance (EPR) and ultraviolet–visible (UV–Vis) spectroscopy) was performed analysing the spectroscopic features of DPPH in mixed ethanol/water solution and the free radical scavenging properties of myrtle leaves extracts and citrus juices. When DPPH was dissolved in mixed solvents, EPR enabled to identify the aggregation phenomena that occur when high amounts of water were employed. On the contrary, UV–Vis revealed only small differences in the absorption maximum among solutions with increasing water contents, without detecting aggregation. EC50 values of myrtle leaf extracts and citrus juices calculated from UV–Vis data were lower than those calculated with EPR. In myrtle extracts, the DPPH depletion measured by UV–Vis was not concentration dependent, revealing the interference in the analysis of the decomposition products of the antioxidants, which absorb at 517xa0nm. EPR spectroscopy was proven to be most reliable with all types of matrix since it is not dependent on the chemical composition of the extract.

Development and Characterization of an Ascorbate Oxidase-based Sensor–Biosensor System for Telemetric Detection of AA and Antioxidant Capacity in Fresh Orange Juice

A new carbon ascorbate oxidase-based sensor-biosensor system (SB) was coupled to a dual-channel telemetric device for online simultaneous electrochemical detection of ascorbic acid (AA) and antioxidant capacity in Hamlin, Sanguinello, and Moro orange varieties. The electrocatalytic performances of the SB were investigated by cyclic voltammetry and amperometric techniques. The phenol composition of orange juice of each variety, and the cyclic voltammetries of the most represented phenols, were provided. The in vitro calibrations were performed in PBS (pH 5.6), applying a constant potential of +500 mV. A standard mixture of phenols, based on orange juice composition, was used as reference material for studying SB behavior. SB works at an applied potential of +500 mV, in a concentration range comprised between the LOD 0.26 μM and 20 μM. In this concentration range, limiting the data acquisition time to 2 min, the problems of electrode passivation due to phenols polymerization were overcome. AA calibration showed that the biosensor registered statistically lower currents than the sensor since the enzyme oxidized AA before it reached the electrode surface. Standard mixture calibration showed that currents registered by sensor and biosensor did not statistically differ. The difference between sensor and biosensor AA registered currents was used to calculate an AA selectivity index and, consequently, to determine the AA content and the antioxidant capacity in the juices. The novelty of the SB is its ability to distinguish between AA and phenols contribution to antioxidant capacity. The obtained results were in accordance with reference methods.

Behavior of the potential antitumor VIVO complexes formed by flavonoid ligands. 3. Antioxidant properties and radical production capability

The radical production capability and the antioxidant properties of some V(IV)O complexes formed by flavonoid ligands were examined. In particular, the bis-chelated species of quercetin (que), [VO(que)2](2-), and morin (mor), [VO(mor)2], were evaluated for their capability to reduce the stable radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) and produce the hydroxyl radical (•)OH by Fenton-like reactions, where the reducing agent is V(IV)O(2+). The results were compared with those displayed by other V(IV)O complexes, such as [VO(H2O)5](2+), [VO(acac)2] (acac=acetylacetonate) and [VO(cat)2](2-) (cat=catecholate). The capability of the V(IV)O flavonoids complexes to reduce DPPH is much larger than that of the V(IV)O species formed by non-antioxidant ligands and it is due mainly to the flavonoid molecule. Through the 5,5-dimethyl-1-pyrroline N-oxide (DMPO) spin trapping assay of the hydroxyl radical it was possible to demonstrate that in acidic solution V(IV)O(2+) has an effectiveness in producing (•)OH radicals comparable to that of Fe(2+). When V(IV)O complexes of flavonoids were taken into account, the amount of hydroxyl radicals produced in Fenton-like reactions depends on the specific structure of the ligand and on their capability to reduce H2O2 to give (•)OH. Both the formation of reactive oxygen species (ROS) under physiological conditions by V(IV)O complexes of flavonoid ligands and their radical scavenging capability can be put in relationship with their antitumor effectiveness and it could be possible to modulate these actions by changing the features of the flavonoid coordinated to the V(IV)O(2+) ion, such as the entity, nature and position of the substituents and the number of phenolic groups.

Influence of pH, buffers and role of quinolinic acid, a novel iron chelating agent, in the determination of hydroxyl radical scavenging activity of plant extracts by Electron Paramagnetic Resonance (EPR)

The Fenton reaction is used to produce hydroxyl radicals for the evaluation of the antioxidant activity of plant extracts. In this paper the parameters affecting the production of hydroxyl radicals and their spin trapping with DMPO were studied. The use of quinolinic acid (Quin) as an Fe(II) ligand was proposed for antioxidant activity determination of Green tea, orange juice and asparagus extracts. Quin, buffers and pH affect the DMPO-OH signal intensity of the EPR spectra. Quin/Fe(II) and low pH enhance the OH generation. Phosphate and Tris-HCl buffers decrease the signal intensity measured in Fe(II)-sulfate and Fe(II)-Quin systems. The extracts were analyzed with Fenton systems containing Fe(II)-sulfate and Fe(II)-Quin with and without buffer. The highest activity was shown with Fe(II)-Quin without buffer, this system being less influenced by pH and chelating agents present in the extracts. This paper will help researchers to better design spin trapping experiments for food matrices.

Microsprinkler irrigation influences clementine fruit quality, leaf temperature and physiology

Abstract The effect on leaf physiology and fruit quality, of two irrigation schedules, daily and twice a week irrigation (control), were tested on ‘SRA 63’ Clementine (Citrus reticulata Blanco) during 2002 and 2003. Micrometeorological data revealed a higher impact of everyday irrigation on air and leaf temperature compared with the control. While in 2002 daily irrigation treatment reduced air temperature up to 3.4°C with respect to the control, in 2003 the cooling effect was greater, with reductions of air temperature up to 9°C as compared with the control. In 2002, gas-exchange related parameters were influenced little by daily undertree microsprinkler irrigation, on the contrary, in 2003 net photosynthetic rates were significantly increased by daily irrigation with respect to the control.

Oxidative stability of plant hydroalcoholic extracts assessed by EPR spin trapping under forced ageing conditions: A myrtle case study

The oxidative stability of myrtle hydroalcoholic extracts was measured, over storage time, with the EPR spin trapping method under forced ageing conditions. The extracts were prepared with 150 and 300u202fgu202fl-1 of berries and extraction media with ethanol ranging from 60 to 90%. Two radicals were detected: the PBN-1-hydroxyethyl adduct and the tert-butyl aminoxyl radical. A dimensionless parameter (Ω) calculated on the basis of the lag time, the rate of formation and concentration of the radical species was used to estimate the extracts oxidative stability. Ω was strongly influenced by the extraction medium, being lower in extracts with ethanol 60%, and by the time of storage. An inverse correlation was calculated between Ω and ellagic acid concentration, thus suggesting the role of this phenolic acid in the antioxidant properties of the extracts. The radical scavenging activity of the extracts against the hydroxyl radical was also measured.