Determination of the antiradical activity and kinetics of pomegranate juice using 2,2-diphenylpicyrl-1-hydrazyl as the antiradical probe.

Whole fruit pomegranate (Punica granatum L.) juice of the 'Wonderful' cultivar was characterized through the elucidation of its antiradical kinetics and activity using 2,2-diphenyl-1-picrylhydrazyl as the antiradical probe. Time-dependent concentration of 2,2-diphenyl-1-picrylhydrazyl during its reduction by the juice has been adjusted through a non-linear parametric fitting. Determined total antiradical activity was high, able to reduce 84.58 µmol/l of 2,2-diphenyl-1-picrylhydrazyl per concentration unit of juice (µl/ml), equivalent to a concentration of 42.29 mmol/l of ascorbic acid (or Trolox). Partial antiradical activities due to the fast-, medium- and slow-kinetics were 49.09, 18.16 and 17.33 µmol/l of reduced 2,2-diphenyl-1-picrylhydrazyl per concentration unit of juice (µl/ml), respectively. The corresponding rate constant for the fast-, medium- and slow-kinetics were κ 1 = 6.03, κ 2 = 0.169 and κ 3 = 0.0094 (µl l)/(ml µmol min), respectively. This methodology allows characterization of samples through the accurate determination of the kinetics of their antiradical features, avoiding the use of empirical approximations that hinder the realistic comparison between extracts independently of their origin.

LC-DAD-ESI/MS(n) determination of direct condensation flavanol-anthocyanin adducts in pressure extracted pomegranate (Punica granatum L.) juice.

Pomegranate (Punica granatum L.) juice, obtained by pressure extraction of the whole fruit, has been analyzed for its flavanol-anthocyanin adduct content using reversed-phase liquid chromatography with diode array detection, coupled to mass spectrometry (ion trap) with electrospray ionization (HPLC-DAD-ESI/MS(n)), operating in positive ion mode. A total of 35 dimers have been detected, consisting of mono- and disubstituted hexoside derivatives of the adducts between the flavan-3-ols (epi)gallocatechin, (epi)catechin and (epi)afzelechin and the anthocyanidins delphinidin, cyanidin and pelargonidin. In addition, evidence is given for the presence of additional anthocyanin-flavanol adducts in this juice.

Reduction kinetics of the antiradical probe 2,2-diphenyl-1-picrylhydrazyl in methanol and acetonitrile by the antiradical activity of protocatechuic acid and protocatechuic acid methyl ester.

This work evaluates the reduction kinetics of the antiradical probe 2,2-diphenyl-1-picrylhydrazyl (DPPH (*)) in methanol and acetonitrile by the antiradical activity of protocatechuic acid (3,4-dihydroxybenzoic acid, 1) and protocatechuic acid methyl ester ( 2). The reduction kinetics of DPPH (*) in both solvents by the antiradical activity of the p-catechol group in 2 is regular, that is, coincide with the proposed standard kinetic model for the reduction kinetics of DPPH (*) by the antiradical activity of an isolated p-catechol group. Therefore, the antiradical activity of 2 experimentally exhibits two rate-two stoichiometric constants in acetonitrile and three rate--three stoichiometric constants in methanol. In contrast, the reduction kinetics of DPPH (*) in both solvents by the antiradical activity of the p-catechol group in 1 is perturbed, that is, deviate from the proposed standard kinetic model. The deviations arise from the presence of the reactive carboxylic acid function which, in methanol, induces an additional reversible side reaction and, in acetonitrile, turns an irreversible reaction reversible, thus modifying the otherwise regular reduction kinetics of DPPH (*) by the antiradical activity of the p-catechol group in 1. On the other hand, the approximated theoretical kinetic equation that applies for those p-catechol groups whose reduction kinetics is regular and that experimentally exhibit three rate--three stoichiometric constants has been derived and used for fitting.

Kinetic model for the antiradical activity of the isolated p-catechol group in flavanone type structures using the free stable radical 2,2-diphenyl-1-picrylhydrazyl as the antiradical probe.

The time evolution of 2,2-diphenyl-1-picrylhydrazyl radical (DPPH*) concentration in four solvents (methanol, ethanol, propanol, and acetonitrile) during its reduction by three flavanones containing an isolated p-catechol group (taxifolin, eriodyctiol, and fustin) as well as the time evolution of the mass spectra of the reaction mixture has been determined by spectrophotometry and liquid mass spectrometry, respectively. In alcoholic solvents the reduction curves consisted of an initial short but fast kinetics step followed by a longer slow kinetics step; in contrast, in acetonitrile the reduction curves completely lacked the slow kinetics step. From the results, a kinetic model for the reaction of reduction of the DPPH* by the isolated p-catechol group in flavanone type structures is proposed. According to this model, the p-catechol group rapidly transfers two hydrogen atoms to DPPH*, through a fast rate constant k1, yielding the corresponding o-quinone. Then, the intermediate o-quinone forms an adduct with the alcoholic solvent, through a slow rate constant k2, and regenerates the p-catechol group. The regenerated p-catechol group reduces additional DPPH* through a fast rate constant k3, yielding the corresponding o-quinone, which can form a new adduct with the solvent to regenerate the p-catechol group, and so on. From the kinetics model, two explicit kinetics equations have been derived that fit very well the experimental data points acquired from all assayed compounds in all of the experiments carried out, thus allowing an accurate determination of the corresponding rate and stoichiometric constants.