The effect of solvent extraction on antioxidant properties of apricot fruit

Two solvent extraction procedures were used to investigate the extraction efficiency in terms of total antioxidant capacity and total phenols in apricot fruit. Samples were either sequentially extracted with aqueous ethanol (ethanol/water 80% v/v) and tetrahydrofuran or directly extracted with tetrahydrofuran. Each extract was analyzed for total antioxidant capacity by the Trolox Equivalent Antioxidant Capacity (TEAC) assay and total phenols by the Folin-Ciocalteu assay. The results showed that using sequential solvent extraction, the majority (85%) of the total antioxidant capacity and total phenols was due to hydrophilic compounds. In tetrahydrofuran direct extractions, the total antioxidant capacity and total phenols were higher than values obtained with aqueous ethanol and the sum of results obtained from sequential extracts for either total antioxidant capacity or total phenols was similar to the tetrahydrofuran-extract antioxidant values. A linear correlation between total antioxidant capacity and total phenols was found and was independent of the solvent extraction method. In conclusion, the choice of solvent is related to the antioxidant potential of fruit and depends on the food hydrophilic/lipophilic composition.     

Redox status of equine seminal plasma reflects the pattern and magnitude of DNA damage in sperm cells

Antioxidant status of seminal plasma from 23 stallions was evaluated. We found a negative correlation between total antioxidant capacity (ABTS^•+ decolorization assay) and thiol content of seminal plasma, and sperm DNA damage (8-oxoG immunostaining, TUNEL reaction, comet assay). Low seminal redox status was the strongest correlated with 8-oxoG level which may indicate that seminal total antioxidant capacity influences mainly the formation of single strand DNA breaks in sperm cells. Since inter-individual differences in seminal antioxidant status were reported, we postulated that the redox status of seminal plasma may be an additional important parameter, both with sperm quantitative and morphological analysis, for evaluation of equine semen quality.     

Antioxidant activity and protective effect on DNA strand scission of Rooibos tea (Aspalathus linearis)

Rooibos tea (Aspalathus linearis) was extracted by refluxing with water and 75% ethanol as a solvent. Antioxidant activity and protective effect on DNA strand scission were investigated by using different antioxidant assay systems and DNA strand nicking assay, respectively. 75% Ethanol extract has higher content of total soluble phenolics and flavonoid than water extract. Antioxidant activities such as hydrogen donating capacity and scavenging activity of hydrogen peroxide were higher in 75% ethanol extract than in water extract except the rate constant with hydroxyl radical. Peroxyl radical induced DNA strand scission was prevented by both 75% ethanol and water extract and hydroxyl radical induced DNA strand scission was not. This result indicates that total soluble phenolics, specially flavonoid, of Rooibos tea are responsible for several kinds of antioxidant activities and preventive activity on peroxyl radical induced DNA strand scission.     

Kinetic analysis of antioxidant capacity of wine

A competition kinetics procedure for measuring total antioxidant capacity in wine is described. This procedure is based on the "crocin bleaching test" [28] as modified for analyzing the antioxidant capacity of complex mixtures [24]. The antioxidant capacity of white wines ranged from 0.08 to 1.2 mM equivalents of the reference antioxidant (Trolox C), while for red wines values ranging from 6.4 to 41.9 mM have been obtained. Although a correlation exists between antioxidant capacity and total phenol content of wines, due to the variable reactivity of different phenol groups, the analysis of the phenol content provides only a crude indication of the actual antioxidant capacity. The analysis of antioxidant capacity on different polyphenol classes, separated by solid phase extraction, indicated that anthocyanins are the major antioxidants of young red wines, and tannins of old red wines and white wines. Several vintages of the same grape have been analyzed and the expected decrease of antioxidant capacity upon ageing was not observed, although spectrophotometric analysis clearly demonstrated the shift from anthocyanin monomers to polymers. Artificial ageing by stirring under air produced a rapid decrease (30 min) of antioxidant capacity followed by an increase (up to two weeks), but not any significant modification of the spectrophotometric chemical age factor. Since neither natural nor artificial ageing present a clear-cut relationship with a decrease of antioxidant capacity, we could conclude that the a priori assumption that an old wine contains less antioxidant capacity, although popular, is not fully correct.     

Pitfalls in a Method for Assessment of Total Antioxidant Capacity

A relatively simple and widely applied method for quantitating the total antioxidant capacity of body fluids and drug solutions based on the absorbance of the ABTS radical cation was evaluated. In this assay, the end-point is an antioxidant-induced decrease in absorbance at a fixed time. This decrease is used as an index of total antioxidant capacity. It is shown that Trolox, potassium cyanide and quercetin all decrease the absorbance of ABTS radical cations at a fixed time, but by different mechanisms. Trolox scavenges the ABTS radical, potassium cyanide inhibits radical formation, while quercetin acts by both mechanisms. Using this method antioxidant capacity may be overestimated, due to both a scavenger effect and an effect on the rate of ABTS oxidation. To distinguish between these effects, a post-addition assay was used in which the sample is added when the formation of radicals is stable. Using post- addition assay conditions enables discrimination between effects on radical scavenging and on the radical formation, two major mechanisms for antioxidant action. In extrapolating the results to an ill vivo situation it should be questioned: (i) whether the peroxidase process does indeed mimic the process of radical formation in vivo, and (ii) whether the ABTS radicals do resemble the radical species involved in an in vivo situation. Results obtained in the ABTS radical-based methods should therefore be reviewed critically before the antioxidant capacity can be assessed.     

Antioxidant capacity and polyphenolic components of teas: implications for altering in vivo antioxidant status

The Oxygen Radical Absorbance Capacity (ORAC) assay was used to determine the total antioxidant capacity of tea. Green and black teas (n = 18) had a mean antioxidant capacity of 761.1 +/- 85.3 micromol Trolox Equivalents (TE) per g dry matter. However, their antioxidant capacity varied from 235 micromol to over 1526 micromol Trolox equivalents (TE)/g dry matter, and total phenolics ranged from 32 to 147 mg/g in different commercial teas. One tea phenolics extract had an antioxidant capacity of 4796 micromol TE/g dry matter and 625 mg total phenolics/g. On a dry matter basis, an antioxidant capacity of 761 micromol TE/g is considerably higher than any of the other fruits and vegetables measured in our laboratory. However, since dry tea is not consumed directly, brewing conditions may influence the final antioxidant capacity in the tea as consumed. We tested both green and black teas by placing one tea bag (1.95 g) in 150 ml (5 oz.) of boiling water. In the first brewed cup, approximately 84% of the total antioxidant activity was solubilized within the first 5 min of brewing. An additional 13% of the antioxidant activity was extracted into the second glass of 150 ml with an additional 5 min of brewing. At the dilutions obtained after the first brewing, the tea as consumed would contain approximately 8. 31 micromol TE per ml. This total antioxidant capacity compares to other drinks from fruits and vegetables that had antioxidant capacity values ranging from 1.6 to 15 micromol TE/ml. At these antioxidant levels, consumption of 150 ml of tea could make a significant contribution to the total daily antioxidant capacity intake. (-)-Epicatechin and (+)-catechin, two components from tea, had an antioxidant capacity of 2.36 and 2.49 micromol/micromol or 8. 13 and 8.58 micromol/mg, respectively. In 16 tea samples we observed a mean of 10.0 +/- 0.6 micromol TE/mg total phenolics. Tea can be an important source of what has been referred to as "non-nutrient" antioxidant phytochemicals. However, with the variation that exists in antioxidant capacity with various tea preparations, measures of antioxidant capacity intake are critical to the study of intake and health outcomes and/or biomarkers of health outcomes.     

Antioxidants in fruits and vegetables: a study of cellular availability and direct effects on human DNA

The effects of several types of whole fruits and vegetables on human lymphocytic DNA were investigated by using two versions of the comet assay. The total antioxidative capacity, as the FRAP value, and ascorbic acid (AA) content were also measured to explore the relationship between the effect and antioxidant content.     

Coffee component 3-caffeoylquinic acid increases antioxidant capacity but not polyphenol content in experimental cerebral infarction

Although coffee has antioxidant capacity, it is not known which of its bioactive compounds is responsible for it, nor has it been analyzed in experimental cerebral infarction. We studied the effect one of its compounds, 3-caffeoylquinic acid (3-CQA), at doses of 4, 25 and 100 μg on plasma antioxidant capacity and plasma polyphenol content, measuring the differences before and after inducing a cerebral infarction in an experimental rat model. We compared them with 3-caffeoylquinic-free controls. The increase in total antioxidant capacity was only higher than in controls in 3-CQA treated animals with the highest dose. This increase in antioxidant capacity was not due to an increase in polyphenols. No differences between the experimental and control group were found regarding polyphenol content and cerebral infarction volume. In conclusion, this increase in antioxidant capacity in the group that received the highest dose of 3-CQA was not able to reduce experimental cerebral infarction.     

Antioxidant status and dialysis: Plasma and saliva antioxidant activity in patients with fluctuating urate levels

The present study is concerned with the influence of processes occurring during dialysis on the antioxidant capacity of plasma and saliva. The biological fluids were also tested for uric acid and total protein content. Before hemodialysis, plasma antioxidant status of hemodialyzed patients appears slightly higher than the corresponding status in normal subjects; after hemodialysis it is found unchanged. The result can be explained by a balance between a reduction in uric acid plasma content, due to the dialytic procedure, and an increase in protein content, possibly due to a dialysis-related hemoconcentration. Moreover, pre-dialysis total antioxidant capacity of whole saliva samples is higher than in healthy individuals and drastically decreases towards normal values following dialytic procedure. Our data indicate a certain concentration of the uric acid in the saliva of hemodialyzed patients and evidence that both total protein concentration and uric acid level show a good correlation with saliva total antioxidant capacity, suggesting that proteins are major antioxidants of this fluid. Further observations are needed to assess whether this improved saliva antioxidant ability has any consequence on the periodontal conditions of hemodialyzed subjects.     

Antioxidants in Fruits and Vegetables: a Study of Cellular Availability and Direct Effects on Human DNA

The effects of several types of whole fruits and vegetables on human lymphocytic DNA were investigated by using two versions of the comet assay. The total antioxidative capacity, as the FRAP value, and ascorbic acid (AA) content were also measured to explore the relationship between the effect and antioxidant content.     

Antioxidant status and dialysis: Plasma and saliva antioxidant activity in patients with fluctuating urate levels

The present study is concerned with the influence of processes occurring during dialysis on the antioxidant capacity of plasma and saliva. The biological fluids were also tested for uric acid and total protein content. Before hemodialysis, plasma antioxidant status of hemodialyzed patients appears slightly higher than the corresponding status in normal subjects; after hemodialysis it is found unchanged. The result can be explained by a balance between a reduction in uric acid plasma content, due to the dialytic procedure, and an increase in protein content, possibly due to a dialysis-related hemoconcentration. Moreover, pre-dialysis total antioxidant capacity of whole saliva samples is higher than in healthy individuals and drastically decreases towards normal values following dialytic procedure. Our data indicate a certain concentration of the uric acid in the saliva of hemodialyzed patients and evidence that both total protein concentration and uric acid level show a good correlation with saliva total antioxidant capacity, suggesting that proteins are major antioxidants of this fluid. Further observations are needed to assess whether this improved saliva antioxidant ability has any consequence on the periodontal conditions of hemodialyzed subjects.     

Tyrosine as important contributor to the antioxidant capacity of seminal plasma

A novel post-addition method, based on the trapping of ABTS-radicals, is applied for studying the total antioxidant capacity of seminal plasma. A remarkable profile is observed, in which seminal plasma quenches radicals in a continuous, relatively slow fashion. Five putative antioxidants present in seminal plasma were studied using the same assay. Some of the compounds such as ascorbic acid, alpha-tocopherol and uric acid exert immediate, fast radical trapping, whereas hypotaurine and tyrosine give rise to the same slow radical trapping curve as seminal plasma. Due to this slow, continuous radical trapping, quantification of the total antioxidant capacity (expressed as trolox equivalent antioxidant capacity, TEAC) strongly depends on the chosen time point after onset of radical trapping. When determined during the slow antioxidant trapping phase, tyrosine has a powerful antioxidant capacity, which in combination with its relatively high plasma concentration makes it an important contributor to the total antioxidant capacity of seminal plasma.     

Antioxidant activity of extracts from leaves and roots of Salvia miltiorrhiza Bunge, S. przewalskii Maxim., and S. verticillata L

Three Salvia species have been studied for antioxidant activity in methanol extracts from roots and leaves. The presence of the polyphenols and tanshinones was screened by HPLC and spectrophotometric assays and related to the antioxidant potential. The antioxidant capacity of the studied species is high, but differences between species and organs have been also revealed. Salvia przewalskii leaf extract was the strongest one in all tests, followed by Salvia miltiorrhiza root and Salvia verticillata leaf. Among the roots, the most active was S. miltiorrhiza extract, followed by S. verticillata. The antioxidant activity correlates to the total polyphenol and, depending on the assay, to the hydroxycinnamic acids content. The high content of tanshinones in both S. miltiorrhiza and S. przewalskii roots is unlikely to contribute to the antioxidant activity.     

Comparison of the contents of the main biochemical compounds and the antioxidant activity of some Spanish olive oils as determined by four different radical scavenging tests

The aim of this study was to compare the contents of the main biochemical compounds and the antioxidant capacity of five Spanish olive oils by four different antioxidant tests and to find out the most valuable oil for disease preventing diets. Fatty acids, sterols and individual antioxidant compounds in Arbequina, Hojiblanca, Extra Virgin, Picual and Lampante Spanish olive oils were determined. Antioxidant activities were done as well using different radical scavenging activities: total radical-trapping antioxidative potential by ABAP (TRAP-ABAP), radical scavenging activity by DPPH (RSA-DPPH), antioxidant assay by beta-carotene-linoleate model system (AA-beta-carotene) and total antioxidant status by ABTS (TAA-ABTS). The highest content of all studied antioxidant compounds (353; 329; 4.6 and 2.7 mg/kg for tocopherols, tocotrienols, polyphenols and o-diphenols, respectively) was found in Extra Virgin oil. Also the highest antioxidant capacity was observed in Extra Virgin oil (668 nM/ml; 29.4%; 40.4% and 2.64 mM TE/kg for TRAP-ABAP, RSA-DPPH, AA- beta-carotene and TAA-ABTS, respectively). The correlation between total phenols and antioxidant capacities measured by four methods was very high, but the highest for the beta-carotene (R = 0.9958). In conclusion, the best method for determination of the antioxidant capacity of olive oils is the beta-carotene test. Extra Virgin olive oil has high organoleptic properties and the highest antioxidant activity. The above-mentioned makes this oil a preferable choice for diseases preventing diets.     

Antioxidant and Antidiabetic Properties of Extracts from Three Underutilized Food Plants of North East India

Three underutilized leafy vegetables Sarcochlamys pulcherrima (Roxb.) Gaudich (SP), Ipomoea aquatica Forssk. (IA) and Zanthoxylum rhetsa (Roxb.) DC (ZR) were extracted with different solvents viz. 95 % ethyl alcohol, methanol and hot water. The extracts were evaluated for their antioxidant potential via DPPH, ABTS and FRAP assay along with electroanalytical studies using cyclic voltammetry. The antidiabetic potential was determined by recording their α-amylase and α-glucosidase inhibitory assay. The total phenolic content (TPC), total flavonoid content (TFC) and the liquid chromatography-mass spectrometry (LC/MS) based phytochemical profiles of the extracts were also determined. All three extracts of SP exhibited significant antioxidant capacity. The antidiabetic potential of the IA and ZR extracts was found to be higher than or at par with that of standard acarbose. LC/MS studies reveal the presence of hitherto reported antioxidant and antidiabetic compounds like gamma-aminobutyric acid, cinnamic acid, caffeic acid, α-viniferin, piperlonguminine, niacin, kaempferol, etc., in the extracts.     

Measuring circulating antioxidants in wild birds

Antioxidants protect against free radical damage, which is associated with various age-related pathologies. Antioxidants are also an important buffer against the respiratory burst of the immune system. This protection presumably has costs and therefore might underlie important life-history trade-offs. Studying such trade-offs in a comparative context requires field-applicable methods for assessing antioxidant capacity in wild animals. Here, we present modifications to a simple spectrophotometric assay (the TEAC or TAS assay) that can be applied to miniscule amounts of blood plasma to determine circulating antioxidant capacity. Additionally, uric acid, the most abundant circulating antioxidant, should be measured independently. Uric acid in birds is derived from amino acid catabolism, perhaps incidentally to its antioxidant function. The assay was validated in experimental studies on chickens showing effects of diet on antioxidant capacity, and in field measurements on 92 species of birds, which demonstrate substantial species differences in constitutive antioxidant capacity. Furthermore, most wild birds demonstrate a dramatic change in antioxidant capacity due to stress. These results show that this technique detects variation appropriate for both interspecific and intraspecific studies, and that antioxidants and uric acid change in response to conditions of interest to field ecologists, such as diet and stress.     

In vivo total antioxidant capacity: comparison of different analytical methods

Several methods have been developed to measure the total antioxidant capacity of a biological sample. The use of peroxyl or hydroxyl radicals as pro-oxidants in the oxygen radical absorbance capacity (ORAC) assay makes it different and unique from the assays that involve oxidants that are not necessarily pro-oxidants. An improvement in quantitation is achieved in the ORAC assay by taking the reaction between substrate and free radicals to completion and using an area-under-curve technique for quantitation compared to the assays that measure a lag phase. The interpretation of the changes in plasma or serum antioxidant capacity becomes complicated by the different methods used in detecting these changes. The interpretation also depends upon the conditions under which the antioxidant capacity is determined because the measurement reflects outcomes in a dynamic system. An increased antioxidant capacity in plasma or serum may not necessarily be a desirable condition if it reflects a response to increased oxidative stress. Similarly, a decrease in plasma or serum antioxidant capacity may not necessarily be an undesirable condition if the measurement reflects decreased production of reactive species. Because of these complications, no single measurement of antioxidant status is going to be sufficient, but a "battery" of measurements, many of which will be described in Forum articles, will be necessary to adequately assess oxidative stress in biological systems.     

Investigations on diverse sesame ( S. indicum L.) germplasm and its wild allies reveal wide variation in antioxidant potential

Free radicals, the key mediators of a range of oxidative reactions involved in lipid oxidation are responsible for food quality deterioration leading to several health hazards. Antioxidants synthesized naturally or synthetically are capable of preventing oxidation of lipids and other related compounds. However, natural antioxidants have many benefits over synthetic ones. Sesame seeds contain large amount of natural bioactive components with high antioxidant potential. In the present study, 14 accessions of sesame containing wild species and cultivars were investigated. The antioxidant potential of sesame seed meal extract was evaluated by total phenolic content (TPC) method using Folin–Ciocalteu reagent, linoleic acid peroxidation by Ferric thiocyanate method, and free radical scavenging assay with 2,2-diphenyl-1-picryl hydrazyl radical. S. laciniatum showed highest mean values for total polyphenol content with maximum % inhibition of linoleic acid peroxidation on 10th day of course of the reaction span and highest antioxidant scavenging power. S. indicum subsp. malabaricum and S. radiatum also showed high total phenol content and radical scavenging capacity. Among the Sesamum indicum cultivars, Gujarat til 2 showed high TPC and high radical scavenging activity. Higher antioxidant property of Sesamum species in comparison to sesame cultivars highlights the need to utilize the wild genepool for the improvement of cultigens for enhanced nutraceutical value.     

First Extensive Polyphenolic Profile of Erodium cicutarium with Novel Insights to Elemental Composition and Antioxidant Activity

Erodium cicutarium is known for its total polyphenolic content, but this work reveals the first highly detailed profile of E. cicutarium, obtained with UHPLC‐LTQ OrbiTrap MS⁴ and UHPLC‐QqQ‐MS/MS techniques. A total of 85 phenolic compounds were identified and 17 constituents were quantified. Overall, 25 new compounds were found, which have not yet been reported for the Erodium genera, or the family Geraniaceae. Along with methanolic extracts, the so far poorly investigated water extracts exhibited in vitro antioxidant activity according to all performed assays, including the ferric reducing/antioxidant power assay (FRAP), 2,2‐diphenyl‐1‐picrylhydrazyl assay (DPPH), 2,2′‐azinobis(3‐ethylbenzthiazoline‐6‐sulfonic acid) assay (ABTS) and cupric ion reducing antioxidant capacity assay (CUPRAC). Elemental composition analysis performed with inductively coupled plasma atomic emission spectrometry (ICP‐AES) and, additionally, hydride generation atomic absorption spectrometry (HydrEA‐ETAAS) showed six most abundant elements to be decreasing as follows: Mg>Ca>K>S>P>Na, and gave first data regarding inorganic arsenic content (109.3–248.4 ng g^?1). These results suggest E. cicutarium to be a valuable source of various phenolic compounds with substantial potential for further bioactivity testing.     

Lymphocyte DNA damage and oxidative stress in patients with iron deficiency anemia

Oxidant stress has been shown to play an important role in the pathogenesis of iron deficiency anemia. The aim of this study was to investigate the association between lymphocyte DNA damage, total antioxidant capacity and the degree of anemia in patients with iron deficiency anemia. Twenty-two female with iron deficiency anemia and 22 healthy females were enrolled in the study. Peripheral DNA damage was assessed using alkaline comet assay and plasma total antioxidant capacity was determined using an automated measurement method. Lymphocyte DNA damage of patients with iron deficiency anemia was significantly higher than controls (p<0.05), while total antioxidant capacity was significantly lower (p<0.001). While there was a positive correlation between total antioxidant capacity and hemoglobin levels (r=0.706, p<0.001), both total antioxidant capacity and hemoglobin levels were negatively correlated with DNA damage (r=-0.330, p<0.05 and r=-0.323, p<0.05, respectively). In conclusion, both oxidative stress and DNA damage are increased in IDA patients. Increased oxidative stress seems as an important factor that inducing DNA damage in those IDA patients. The relationships of oxidative stress and DNA damage with the severity of anemia suggest that both oxidative stress and DNA damage may, in part, have a role in the pathogenesis of IDA.