Loading of free radicals on the functional graphene combined with liquid chromatography–tandem mass spectrometry screening method for the detection of radical-scavenging natural antioxidants

A novel free radical reaction combined with liquid chromatography electrospray ionization tandem mass spectrometry (FRR-LC–PDA-ESI/APCI-MS/MS) screening method was developed for the detection and identification of radical-scavenging natural antioxidants. Functionalized graphene was prepared by chemical method for loading free radicals (superoxide radical, peroxyl radical and PAHs free radical). Separation was performed with and without a preliminary exposure of the sample to specific free radicals on the functionalized graphene, which can facilitate reaction kinetics (charge transfers) between free radicals and potential antioxidants. The difference in chromatographic peak areas is used to identify potential antioxidants. The structure of the antioxidants in one sample (Swertia chirayita) is identified using MS/MS and comparison with standards. Thirteen compounds were found to possess potential antioxidant activity, and their free radical-scavenging capacities were investigated. The thirteen compounds were identified as 1,3,5-trihydroxyxanthone-8-O-β-d-glucopyranoside (PD1), norswertianin (PD2), 1,3,5,8-tetrahydroxyxanthone (PD3), 3, 3′, 4′, 5, 8-penta hydroxyflavone-6-β-d-glucopyranosiduronic acid-6′-pentopyranose-7-O-glucopyranoside (PD4), 1,5,8-trihydroxy-3-methoxyxanthone (PD5), swertiamarin (PS1), 2-C-β-d-glucopyranosyl-1,3,7-trihydroxylxanthone (PS2), 1,3,7-trihydroxylxanthone-8-O-β-d-glucopyranoside (PL1), 1,3,8-trihydroxyl xanthone-5-O-β-d-glucopyranoside (PL2), 1,3,7-trihydroxy-8-methoxyxanthone (PL3), 1,2,3-trihydroxy-7,8-dimethoxyxanthone (PL4), 1,8-dihydroxy-2,6-dimethoxy xanthone (PL5) and 1,3,5,8-tetramethoxydecussatin (PL6). The reactivity and SC₅₀ values of those compounds were investigated, respectively. PD4 showed the strongest capability for scavenging PAHs free radical; PL4 showed prominent scavenging capacities in the lipid peroxidation processes; it was found that all components in S. chirayita exhibited weak reactivity in the superoxide radical scavenging capacity. The use of the free radical reaction screening method based on LC–PDA-ESI/APCI-MS/MS would provide a new approach for rapid detection and identification of radical-scavenging natural antioxidants from complex matrices.     

Novel Kinetic Method for Expressing the Ability of Antioxidant to Scavenge Radicals

A method was developed to treat the result from an antioxidant trapping radicals including 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonate) cationic radical(ABTS^+·), 2,2'-diphenyl-1-picrylhydrazyl radical(DPPH), and galvinoxyl radical. In the presence of a certain concentration of an antioxidant, the decrease of the concentration of a kind of radicals follows the second order exponential function with the increase of the reaction time(t), viz.,[radical]=Ae^-t/a+Be^-t/b+C, the derivation operation of which obtains the differential style, -d[radical]/dt= (A/a)e^-t/a+(B/b)e^-t/b, revealing the relationship between the reaction rate(r=-d[radical]/dt) and the reaction time(t). Thus, the reaction rate at the beginning of the reaction(r₀) can be calculated by assigning t=0 in the equation of -d[radical]/dt-t. Based on the concept of the reaction rate, r=k[radical][antioxidant], the rate constant(k) can be calculated based on r₀ and the initial concentrations of radical and antioxidant, k=r₀/([radical]₀[antioxidant]₀). The k means the rate of a fresh antioxidant molecule to trap a fresh radical. This method was used to treat the interactions of ABTS^+·, DPPH, and galvinoxyl radicals with three homoisoflavonoids, four pyrazoles, and three ferrocenyl Schiff bases. It was found that ferrocenyl group is beneficial for antioxidants to reduce ABTS^+·, and ortho-hydroxyl group is beneficial for antioxidants to donate hydrogen atom in phenolic hydroxyl group to DPPH and galvinoxyl radicals.     

Radical-scavenging capacity of phenol fractions in the brown seaweed Ascophyllum nodosum: an electrochemical approach

In this article, the radical-scavenging capacity of phenol fractions extracted from the brown seaweed Ascophyllum nodosum was assessed using in parallel colorimetric methods (ABTS and DPPH) and electrochemistry (cyclic voltammetry). Results obtained by the three methods correlated in the case of global fractions, whereas only ABTS and DPPH correlated when activities were expressed on a phenol basis. The successive fractions separated by both their average molecular size and their polarity exhibited activities largely dependant on their phenol content, suggesting that phlorotannins are the main anti-oxidant molecules in hydro-alcoholic extracts of A. nodosum. In addition, phenol fractions of relative low molecular weight were clearly more active than others. This work opens new opportunities to better evaluate the radical-scavenging potential of phenol pools in algae using both bi-parametric fractionating and electrochemistry.     

Synthesis of a new stable and water-soluble tris(4-hydroxysulfonyltetrachlorophenyl)methyl radical with selective oxidative capacity

A new stable organic free radical of the PTM (perchlorotriphenylmethyl) series very soluble in water is reported. This free radical is sensitive to electron transfer processes, and the selectivity of these reactions in the presence of ascorbic acid, pyrogallol, and catechol as reducing species is described. The electron paramagnetic resonance spectrum and the electrochemical behavior are also presented.