Stanislava Gorjanović

Application of a Novel Antioxidative Assay in Beer Analysis and Brewing Process Monitoring

A novel antioxidative assay based on direct current polarography has been developed. Quantification of antioxidative (AO) activity has been based on a decrease of hydrogen peroxide anodic current in the presence of antioxidants. An efficient experimental procedure, without any special pretreatment of analyzed samples, has been applied. Antioxidative activity of different kinds of commercial beers (dark, blond, and alcohol-free), some small-scale made special beers with medicinal herbs and mushroom extracts, extracts themselves, as well as individual phenolic components present in beer has been measured. In addition, changes of AO activity during the full-scale industrial process of beer production have been monitored. A strong correlation between results obtained and total phenolics content has been observed. The assay can be recommended for application in brewing industry, either to survey a process with the aim to optimize relevant technological factors or to analyze quality of final product.

Antioxidant activity of wines determined by a polarographic assay based on hydrogen peroxide scavenge.

Antioxidant (AO) activity of various red and white wines of different origin as well as some individual phenolic compounds present in wine has been assessed using a polarographic assay. Direct current polarography has been used to survey hydrogen peroxide scavenge (HPS) upon gradual addition of tested samples. Results expressed as reciprocal value of wine volume required for 50% decrease of anodic limiting current of hydrogen peroxide have been validated through correlation with Folin-Ciocalteau and DPPH assays. All wines exhibit HPS activity analogous with total phenolic content and DPPH scavenge. Reliability and accuracy, low cost, and rapid and direct experimental procedure open a wide area for application of this assay, making it a good alternative to standard, widely accepted AO assays.

Polarographic study of hydrogen peroxide anodic current and its application to antioxidant activity determination.

Behavior of hydrogen peroxide in alkaline medium has been studied by direct current (DC) polarography with dropping mercury electrode (DME) aiming to apply it in antioxidant (AO) activity determination. Development of a peroxide anodic current having form of a peak, instead of common polarographic wave, has been investigated. As a base for this investigation the interaction of H(2)O(2) with anodically dissolved mercury was followed. Formation of mercury complex [Hg(O(2)H)(OH)] has been confirmed. The relevant experimental conditions, such as temperature, concentration and pH dependence, as well as time stability of hydrogen peroxide anodic current, have been assessed. Development of an AO assay based on decrease of anodic current of hydrogen peroxide in the presence of antioxidants (AOs) has been described. Under optimized working conditions, a series of benzoic acids along with corresponding cinnamate analogues have been tested for hydrogen peroxide scavenging activity. In addition, the assay versatility has been confirmed on various complex samples.

Changes of hydrogen peroxide and radical-scavenging activity of raspberry during osmotic, convective, and freeze-drying.

This study was conducted to investigate the influence of different drying treatments on antioxidant (AO) activity and phenolic content of raspberry (Rubus idaeus), cultivar Willamette. Whole raspberry fruits were dried convectively (air-drying), osmotically, and freeze-dried. Acetone-water extracts of fresh and dried raspberries were assessed for total phenolic content by standard Folin-Ciocalteau method. Two AO assays were applied, a recently developed direct current (DC) polarographic assay based on decrease of anodic oxidation current of hydrogen peroxide and widely used radical scavenge against the 1,1-diphenyl-2-picrylhydrazyl (DPPH). Strong correlation has been obtained between both AO assays and total phenolic content. In addition, some individual phenolic compounds present in raspberry have been assessed using DPPH and DC polarographic assay. Comparison and evaluation of drying methods has been based on preservation of AO activity and total phenolic content. Obtained results confirmed superiority of freeze-drying; convective drying caused slight changes while osmotic dehydration showed a significant decrease of phenolic compounds and AO activity.

Antioxidant activity of propolis extracts from Serbia: A polarographic approach

Antioxidant activity (AO) of commercial propolis extracts (PEs), available on Serbian market, was determined by direct current (DC) polarography. Polarographic anodic current of 5.0 mmol L(-1) alkaline solution of H2O2 was recorded at potentials of mercury dissolution. Decrease of the current was plotted against the volume of gradually added PEs. The volume of PE causing 20% current decrease was determined from the linear part of the plot. Antioxidant activity was expressed in H2O2 equivalent (HPEq), representing the volume of PE that corresponds to 1.0 mmol L(-1) H2O2 decrease. Resulting HPEq ranged between 1.71±0.11 and 8.00±0.18 μL. Range of 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging activity was from 0.093±0.004% to 0.346±0.006%. Total phenolic content (TCP) of PE with superior AO activity was 5.31±0.05% g GAE, while the extract with the lowest activity contained 1.45±0.02% g GAE. Antioxidant activity, determined by polarographic method, was correlated with DPPH scavenging activity (R2=0.991) and TCP (R2=0.985). Validity of obtained results was further confirmed using ANOVA and post hoc Tukey HSD test.

Electrochemical versus spectrophotometric assessment of antioxidant activity of hop (Humulus lupulus L.) products and individual compounds.

Antioxidant (AO) activity of extracts of hop cones (Serbian domestic varieties) and commercial hop products (Saaz, Spalter, Spalter select, and Magnum pellets) was determined by parallel application of recently developed direct current (DC) polarographic and widely used DPPH assay. Correlations between 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) scavenging and total phenolics (TPC) determined by the Folin-Ciocalteu assay (FC) (0.99), and between H2O2 scavenging (HPS) and humulone content (H) determined by conductometric method (0.94), total resins (TR) (0.85), and hop storage index (HIS) (-0.90), were found statistically significant at p < 0.05 level while complete lack of HPS correlation with TPC and DPPH was observed. To obtain an insight into differences between results of AO assays applied, activity of individual compounds, prevalent hop phenolics, and bitter acids was determined. By far superior HPS activity of humulone was followed by catechin, quercetin, xanthohumol, lupulone, and rutin. In contrast, DPPH scavenging activity of phenolics (quercetin > catechin > rutin > xantohumol) was found substantially higher than activity of bitter acids. According to ferric reducing antioxidant power (FRAP) and scavenging of 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), higher AO activity was ascribed to phenolics, while almost neglecting humulone. Besides reliability, low cost, and an easy-to-handle procedure, an ability to recognize humulone as the major contributor of hop AO activity could allow DC polarographic assay to be applied in analysis of various hop-derived products.

Isolation and Characterization of Highly Liganded Protein from Brewer's Barley Grain

Two basic proteins, HLP-1 and HLP-2, were isolated from brewers barley grain (Hordeum vulgare L.) and characterized as glycoproteins with molecular masses of 16 and 13 kDa and pI values of 7.4 and 8.8, respectively. They could bind sugars, metal ions, and both hydrophobic and hydrophylic molecules of low molecular mass. These characteristics may be related to their potential plant-protecting role.

Trametes versicolor ethanol extract, a promising candidate for health–promoting food supplement

Abstract This study aimed to estimate antiradical, antioxidant (AO) and cytotoxic activities of the fungus Trametes versicolor ethanol fruiting body extract. The extract was found to effectively scavenge both O2•− and NO• (29.62 and 52.48 μg/mL, respectively). It also showed a good AO activity in the polarographic HPMC assay (950%/mL). p-Hydroxybenzoic acid may be one of the responsible compounds for the afore-mentioned activities. The same extract also exhibited a concentration-dependent cytotoxicity against MCF-7 and HepG2 tumour cell lines reaching IC50 values of 123.51 and 134.29 μg/mL, respectively with no cytotoxic activity against normal MRC-5 cells. Gentisic, syringic and protocatechuic acids may be among the bioactive principles for the observed cytotoxicity. Taken all together, T. versicolor ethanol extract can be considered as a promising candidate for development of health promoting food supplement.

Antioxidant efficiency of polyphenols from coffee and coffee substitutes-electrochemical versus spectrophotometric approach

Antioxidant (AO) capacity of instant, espresso, filter and Turkish/Greek coffee brews, coffee substitutes (roasted chicory root, barley, pea, chickpea, carob and dried fig) and individual compounds (phenolic acids, flavonoids, methylxanthines, N-methyl pyridinium and HMW melanoidins) was assessed using DC polarographic assay based on decrease of anodic current originating from hydroxo-perhydroxo mercury complex formed in alkaline solutions of H2O2 at potential of mercury dissolution, as well as three spectrophotometric assays (DPPH, ABTS and FRAP). A large difference between applied assays ability to recognize various types of individual AOs was noticed. Only according to DC polarographic assay significant AO activity was ascribed to methylxanthines and N-methyl pyridinum. The total content of phenolics (TPC) present in complex samples was determined by FC assay. The highest TPC was ascribed to instant coffees and coffee substitutes while the lowest to decaffeinated filter coffee. Complex samples were grouped based on principal components analysis, phenolics AO coefficient, calculated as the ratio between AO capacity and TPC, and relative AO capacity index (RACI), calculated by assigning equal weight to all applied assays including FC. The highest values of RACI were ascribed to instant coffee brews, followed by substitutes while the lowest to the decaffeinated espresso coffee.

Antioxidant Capacity Determination of Complex Samples and Individual Phenolics - Multilateral Approach

Antioxidant (AO) capacity of various medicinal plants extracts and phenolic compounds was assessed by the most widely used spectrophotometric assays such as ferric reducing antioxidant power (FRAP) and scavenging of 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH). In addition, two direct current (DC) polarographic assays, one based on a decrease of anodic current of [Hg(O2H)(OH)] - HydroxoPerhydroxoMercury(II) Complex (HPMC) formation in alkaline solution of H2O2, at the potential of mercury dissolution and another recently developed Mercury Reduction Antioxidant Power (MRAP), based on a a decrease of cathodic current of Hg(II) reduction were employed. Percentage of both currents decrease was plotted versus the volume of gradually added complex samples or the amount of individual ones and the slopes of these plots were used to express AO capacity. Total phenolic content (TPC) of extracts was determined by Folin- Ciocalteu (FC) assay. Correlations between applied assays were calculated by regression analysis. Relative Antioxidant Capacity Index (RACI), calculated by assigning equal weight to all applied assays and Phenolic Antioxidant Coefficients (PAC), calculated as a ratio between particular AO capacity and TPC, were used to achieve more comprehensive comparison between analyzed samples, as well as applied assays.