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D����������������Total Antioxidant Capacity and phenolic compounds of selected vinegars in the Greek Market.

Katerina P. Skenderi1, Ilias Haligiannis2, Nikolaos M. Sitaras2

1Laboratory of Nutrition and Clinical Dietetics, Department of Nutrition and Dietetics, Harokopio University, Athens, Greece 
2Department of Pharmacology, Medical School, University of Athens, Athens, Greece

Address for correspondence:	
Katerina P. Skenderi, Ph.D., 
Laboratory of Nutrition and Clinical Dietetics, Department of Nutrition and Dietetics, Harokopio University, Athens, Greece
Tel.: +30 2109549177; 
Fax: +30 2109549141; 
E-mail:  HYPERLINK "mailto:ksken@hua.gr" ksken@hua.gr
1. Introduction
Epidemiological and clinical studies have demonstrated that consumption of foods rich in antioxidant compounds has beneficial effects on human health and protects against a great variety of diseases  ADDIN EN.CITE  ADDIN EN.CITE.DATA [1-2]. Oxidative stress, which is responsible for molecular and cellular damages, is also believed to be the major cause of many degenerative diseases as cardiovascular diseases or cancer. The detrimental effects of oxidative stress are scavenged by the antioxidant capacity of the organism, which is of endogenous or exogenous nature. The exogenous antioxidants consist of nutrients and derive from foods or beverages we consume. The most famous foods, which have an important role in maintaining this equilibrium, are fruits, vegetables or their by-products  ADDIN EN.CITE  ADDIN EN.CITE.DATA [3-5].
Measurement of antioxidant capacity of different foods lightens up the importance of nutrition against free radicals. Vitamins such as vitamin C, vitamin A, vitamin E and also minerals such as selenium and zinc are very important nutrients to maintain the antioxidant capacity of an organism. Additionally, a variety of other substances such as carotenoids, flavonoids, polyphenols, phenolic acid and uric acid are very famous antioxidants  ADDIN EN.CITE  ADDIN EN.CITE.DATA [6]. Among these substances, polyphenols and their by-products are the most important antioxidants in plants. Several studies in humans or in animals have demonstrated that polyphenols possess significant chemopreventive properties due to their antioxidant capacity  ADDIN EN.CITE  ADDIN EN.CITE.DATA [6-7]. Polyphenols are present in fruits, wines and their by-products like vinegars. Although the antioxidant role of many fruits and different wines in vitro and in vivo upon healthy and different kind of disease subjects has been extensively studied  ADDIN EN.CITE  ADDIN EN.CITE.DATA [8-10], there are fewer studies to examine the antioxidant activity of vinegars.
Vinegars are widely used as acidic seasoning and are commercially grouped as wine vinegars, red or white, and fruit vinegars. Balsamic vinegars are products of grape must with a specific fermentation and ageing procedure. Vinegars contain different antioxidant compounds and their final quality depends to the raw material used as substrate, the acetification system used and the ageing procedure used during their production  ADDIN EN.CITE <EndNote><Cite><Author>Tesfaye</Author><Year>2002</Year><RecNum>44</RecNum><record><rec-number>44</rec-number><foreign-keys><key app="EN" db-id="ze50tdz2i2vsfjewxwaxeazof2zz2zdvpawv">44</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Tesfaye, W.</author><author>Morales, M. L.</author><author>Garcia-Parrilla, M. C.</author><author>Troncoso, A. M.</author></authors></contributors><auth-address>Area de Nutricion y Bromatologia, Facultad de Farmacia, Universidad de Sevilla, C/ P. Garcia Gonzalez s/n, E-41012 Sevilla, Spain.</auth-address><titles><title>Evolution of phenolic compounds during an experimental aging in wood of Sherry vinegar</title><secondary-title>J Agric Food Chem</secondary-title></titles><periodical><full-title>J Agric Food Chem</full-title></periodical><pages>7053-61</pages><volume>50</volume><number>24</number><edition>2002/11/14</edition><keywords><keyword>Acetic Acid/*chemistry</keyword><keyword>Aldehydes/analysis</keyword><keyword>Chemistry, Physical</keyword><keyword>Chromatography, High Pressure Liquid</keyword><keyword>Coumaric Acids/analysis</keyword><keyword>Discriminant Analysis</keyword><keyword>Ethanol/analysis</keyword><keyword>Furaldehyde/analysis</keyword><keyword>Hydrogen-Ion Concentration</keyword><keyword>Hydroxybenzoic Acids/analysis</keyword><keyword>Phenols/*analysis</keyword><keyword>Physicochemical Phenomena</keyword><keyword>Quercus</keyword><keyword>Time Factors</keyword><keyword>*Wine</keyword><keyword>*Wood</keyword></keywords><dates><year>2002</year><pub-dates><date>Nov 20</date></pub-dates></dates><isbn>0021-8561 (Print)&#xD;0021-8561 (Linking)</isbn><accession-num>12428959</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=12428959</url></related-urls></urls><electronic-resource-num>jf020602x [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>[11].
Due to the chemical diversity of antioxidant compounds present in vinegars and perhaps the additive, synergistic and antagonistic interactions among these molecules and also the other nutrients present in them, do not necessary reflect their total antioxidant capacity (TAC)  ADDIN EN.CITE  ADDIN EN.CITE.DATA [7, 12]. Therefore, the knowledge of vinegars Total Antioxidant Capacity would be interesting and useful. For this purpose, several methods have been developed for measuring the TAC of food and beverages. Among these methods, the automated photometric assay BAP (Biological Antioxidant Potential)  ADDIN EN.CITE <EndNote><Cite><Author>Pasquini</Author><Year>2008</Year><RecNum>66</RecNum><record><rec-number>66</rec-number><foreign-keys><key app="EN" db-id="ze50tdz2i2vsfjewxwaxeazof2zz2zdvpawv">66</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Pasquini, A.</author><author>Luchetti, E.</author><author>Marchetti, V.</author><author>Cardini, G.</author><author>Iorio, E. L.</author></authors></contributors><auth-address>Department of Veterinary Clinics, University of Pisa, V.le delle Piagge, 2, 56124 Pisa, Italy. a.pasquini@vet.unipi.it</auth-address><titles><title>Analytical performances of d-ROMs test and BAP test in canine plasma. Definition of the normal range in healthy Labrador dogs</title><secondary-title>Vet Res Commun</secondary-title></titles><periodical><full-title>Vet Res Commun</full-title></periodical><pages>137-43</pages><volume>32</volume><number>2</number><edition>2007/09/12</edition><keywords><keyword>Animals</keyword><keyword>Antioxidants/*metabolism</keyword><keyword>Dogs/*blood</keyword><keyword>Female</keyword><keyword>*Health</keyword><keyword>Male</keyword><keyword>Reactive Oxygen Species/*blood</keyword><keyword>Reference Values</keyword><keyword>Reproducibility of Results</keyword></keywords><dates><year>2008</year><pub-dates><date>Feb</date></pub-dates></dates><isbn>0165-7380 (Print)&#xD;0165-7380 (Linking)</isbn><accession-num>17846911</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=17846911</url></related-urls></urls><electronic-resource-num>10.1007/s11259-007-9014-x</electronic-resource-num><language>eng</language></record></Cite></EndNote>[13] measures the ferric reducing antioxidant power of the sample.
The aim of our study was to measure the TAC of 27 different vinegars, purchased from a local market, using the BAP test and also to check their TAC�s stability after a three month period, kept under housekeeping conditions. 

2. Materials and methods
2.1 Samples analysed
Twenty-seven vinegars were purchased from a local super-market and were grouped in 12 wine, 9 balsamic and 6 fruit vinegars, with regard to the information provided on the bottle label, their origin and their characteristics, as shown on table 1. Seventeen of them were on sale in glass containers and 10 in plastic containers. Furthermore, vinegars were subgrouped in 4 groups depending of their colour and their variety, 17 red, 4 white, 4 grape and 2 apple vinegars, also presented in table 1.
All samples were first filtered (0.5mm filter) to remove any solid material, and then were diluted 5-10 folds in dis-distilled water to fit within the linearity range (CV 400-10000�mol Fe2+) of the method. Following the first measurement, all vinegars were kept in a dry place at room temperature for 3 months, in order to expose them at normal housekeeping conditions and then experiments were repeated. All samples were assayed in triplicate. Acidity, sugar content, total phenolic content and pH were also recorded.

2.2 Total Antioxidant Capacity Measurement
The total antioxidant capacity was measured by BAP test in an automated photometric assay which can be carried out by the analytical system photometer F.R.E.E (Free Radical Elective Evaluator, DIACRON International S.r.I., Grosseto Italy) at 505 nm wavelength with optical path 1 cm after incubation of the sample and reagents at 37 oC in the thermostatic blocks of the photometer. 
Briefly, BAP test is based on the ability of a coloured solution, containing ferric ions (Fe3+) adequately bound to a special chromogenic substrate, to decolour when its Fe3+ ions are reduced to ferrous ions (Fe2+) as well as it can be observed by adding a reducing system, i.e vinegar sample.
Therefore, in the BAP test, the sample to be tested is dissolved in a coloured solution that has been previously obtained by mixing a source of ferric ions (FeCl3, ferric chloride) with a special chromogenic substrate (a thiocyanate-derived compound). After five minutes incubation, such solution will decolour and the intensity of this change will be directly proportional to the ability of our vinegar to reduce ferric ions. By assessing photometrically the intensity of decolouration, the amount of reduced ferric ions can be calculated and the reducing ability or antioxidant capacity of our sample tested can be measured. The BAP test provides a global measurement of several antioxidants, including ascorbic acid, a-tocopherol, uric acid and bilirubin  ADDIN EN.CITE <EndNote><Cite><Author>Benzie</Author><Year>1996</Year><RecNum>67</RecNum><record><rec-number>67</rec-number><foreign-keys><key app="EN" db-id="ze50tdz2i2vsfjewxwaxeazof2zz2zdvpawv">67</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Benzie, I. F.</author><author>Strain, J. J.</author></authors></contributors><auth-address>Department of Health Sciences, Hong Kong Polytechnic University, Kowloon, Hung Hom, Hong Kong.</auth-address><titles><title>The ferric reducing ability of plasma (FRAP) as a measure of &quot;antioxidant power&quot;: the FRAP assay</title><secondary-title>Anal Biochem</secondary-title></titles><periodical><full-title>Anal Biochem</full-title></periodical><pages>70-6</pages><volume>239</volume><number>1</number><edition>1996/07/15</edition><keywords><keyword>Adult</keyword><keyword>Aged</keyword><keyword>Animals</keyword><keyword>Antioxidants/*analysis</keyword><keyword>Cattle</keyword><keyword>Colorimetry/*methods</keyword><keyword>Dose-Response Relationship, Drug</keyword><keyword>Ferric Compounds/*chemistry</keyword><keyword>Ferrous Compounds/*chemistry</keyword><keyword>Humans</keyword><keyword>Indicators and Reagents</keyword><keyword>Kinetics</keyword><keyword>Middle Aged</keyword><keyword>Oxidation-Reduction</keyword><keyword>Plasma/*metabolism</keyword><keyword>Reproducibility of Results</keyword><keyword>Triazines/chemistry</keyword></keywords><dates><year>1996</year><pub-dates><date>Jul 15</date></pub-dates></dates><isbn>0003-2697 (Print)&#xD;0003-2697 (Linking)</isbn><accession-num>8660627</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=8660627</url></related-urls></urls><electronic-resource-num>S0003-2697(96)90292-4 [pii]&#xD;10.1006/abio.1996.0292</electronic-resource-num><language>eng</language></record></Cite></EndNote>[14].
The biological antioxidant potential (BAP) is expressed as �mol Fe2+/L. The optimum value of the normal human plasma antioxidant potential, has been defined to be >2200 �mol/L of reduced iron.

2.3 Total phenols
Total phenols which are present in vinegars were determined by the Folin-Ciocalteu method  ADDIN EN.CITE <EndNote><Cite><Author>Singleton</Author><Year>1965</Year><RecNum>53</RecNum><record><rec-number>53</rec-number><foreign-keys><key app="EN" db-id="ze50tdz2i2vsfjewxwaxeazof2zz2zdvpawv">53</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Singleton, V.L., and Rossi, J.A.</author></authors></contributors><titles><title>Colorimetry of total phenolics and phosphomolybdic-phosphotungstic acid reagents.</title><secondary-title>American Journal of Enology and Viticulture</secondary-title></titles><periodical><full-title>American Journal of Enology and Viticulture</full-title></periodical><pages>144-158</pages><volume>16</volume><dates><year>1965</year></dates><urls></urls></record></Cite></EndNote>[15] by doing some modification to reduce the assay volume. In a solution of 3.90 ml H2O and 0.1 ml sample, 0.5 ml Folin-Ciocalteu reagent mix (25 ml Folin-Ciocalteu reagent (Merck, Darmstadt, Germany) in 75 ml H2O) was added. After 5 minutes, 0.5 ml saturated sodium carbonate (20% w/v) was added. After vigorous vortexing, the final solution was stand at room temperature for 30 min. The reading was performed at 725nm (Cecil spectrophotometer, 1000 series, Cecil Instruments Ltd, Cambridge, England). The results were expressed as gallic acid equivalents per liter of sample (GAE/l) using a calibration curve against a gallic acid standard (50-500mg/l ADDIN EN.CITE <EndNote><Cite><Author>Singleton</Author><Year>1965</Year><RecNum>53</RecNum><record><rec-number>53</rec-number><foreign-keys><key app="EN" db-id="ze50tdz2i2vsfjewxwaxeazof2zz2zdvpawv">53</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Singleton, V.L., and Rossi, J.A.</author></authors></contributors><titles><title>Colorimetry of total phenolics and phosphomolybdic-phosphotungstic acid reagents.</title><secondary-title>American Journal of Enology and Viticulture</secondary-title></titles><periodical><full-title>American Journal of Enology and Viticulture</full-title></periodical><pages>144-158</pages><volume>16</volume><dates><year>1965</year></dates><urls></urls></record></Cite></EndNote>[15]).

2.4 Colour measurements
The colour characteristics of vinegars were determined using spectrophotometric measurements at A420nm, A520nm, and A620nm. Colour density IC was calculated as the sum of A420nm and A520nm. Tint T was calculated as the ratio of A420nm to A520nm. Colour intensity IC� was determined as the sum of A420nm, A520nm and A620nm. dA(%) was calculated as {[A520nm-(A420nm-A620nm)/2]x100}/A520nm. K-K value was found as the log value of the A420nm and A520nm sum. Proportions of red (%R), yellow (%Y) and blue (%B) were determined as A520nm x 100/IC�, and A420nm x 100/IC�, and A620nm x 100/IC�, respectively  ADDIN EN.CITE <EndNote><Cite><Author>Yildirim</Author><Year>2006</Year><RecNum>48</RecNum><record><rec-number>48</rec-number><foreign-keys><key app="EN" db-id="ze50tdz2i2vsfjewxwaxeazof2zz2zdvpawv">48</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Yildirim, Hatice Kalkan</author></authors></contributors><titles><title>Evaluation of colour parameters and antioxidant activities of fruit wines</title><secondary-title>International Journal of Food Sciences and Nutrition</secondary-title></titles><periodical><full-title>International Journal of Food Sciences and Nutrition</full-title></periodical><pages>47-63</pages><volume>57 (1/2)</volume><section>47</section><dates><year>2006</year></dates><urls></urls></record></Cite></EndNote>[16].

2.5 Sugar content
The sugar content (%) of vinegars was determined by using a sugar refractometer (Mod.300003, Sper Scientific Ltd, NY, USA). The refractometer works using the principle of light refraction through liquids. As light passes from air into a liquid it slows down. This phenomenon is what gives a �bent� look to objects partially submerged in water. Refractometers measure the amount of dissolved solids in liquids by passing through a sample and showing the refracted angle on a scale. The scale is defined as the number of grams of pure sugar dissolved in 100 ml of pure water (grams of sugar/100 ml H2O)  ADDIN EN.CITE <EndNote><Cite><Author>VanWaes</Author><Year>1998</Year><RecNum>50</RecNum><record><rec-number>50</rec-number><foreign-keys><key app="EN" db-id="ze50tdz2i2vsfjewxwaxeazof2zz2zdvpawv">50</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>C. VanWaes </author></authors></contributors><titles><title><style face="normal" font="default" size="100%">A Rapid Determination of the Total Sugar Content and the Average Inulin Chain Length in Roots of Chicory (</style><style face="italic" font="default" size="100%">cichorium intybus L</style><style face="normal" font="default" size="100%">)</style></title><secondary-title>J Sci Food Agric</secondary-title></titles><periodical><full-title>J Sci Food Agric</full-title></periodical><pages>107-110</pages><volume>76</volume><dates><year>1998</year></dates><urls></urls></record></Cite></EndNote>[17].

2.6 Statistical analysis
Results were analysed with SPSS 19 software and were expressed as mean�SD. The normality was tested by performing the Kolmogorov � Smirnov test. Due to lack of normality the variables are presented as median, quartiles and min-max. The Kruskal Wallis non parametric ANOVA test was used to evaluate statistically significant differences between groups. Correlations were performed using the Spearman�s rank order coefficient and pair wise differences were evaluated using the Wilcoxon test. Changes over time for overall twelve parameters (antioxidant activity, A420, A520, A620, IC, IC�, T, dA%, K-K, %Red, % Yellow, % Blue) were studied using Signed Ranks test.

3. Results and discussion

The main characteristics of vinegars and their brand names are presented in table 1. Their colour parameters and their antioxidant potential (BAP) at time 0 (1st measurement) are presented in table 2 and the same parameters measured after a three month period are shown in table 3. Among the first main groups (wine, balsamic and fruit vinegars), balsamic vinegars demonstrated statistically higher BAP values compared to both wine and fruit vinegars (p <0.001 and p=0.006, respectively) (Table 4). 
Antioxidant capacity was found to be positively correlated with both, sugar and phenol content of vinegars (figures 1a-b). Additionally, BAP values exhibited a positive and statistically significant correlation with IC, IC� and K-K values (figures 2a-c), respectively. 
Each group of vinegars (balsamic, fruit, wine, red, white, apple and grape) was studied for BAP changes over a three month period. The absence of any statistically significant difference among any group confirmed stability of vinegars� antioxidant capacity under housekeeping conditions for up to this period.  However, analysis of colour characteristics of vinegars revealed that time can affect vinegars regarding those colour characteristics, with IC, IC� and K-K value being significantly lower after three months (figure 3a-c). After the three month period, there was a decrease in IC (p=0.001), in IC� (p=0.001) and K-K (p=0.003).
The present study was aimed to elucidate the antioxidant activity of the most popular vinegars in the Greek market, and to highlight the quality of this seasoning product in the Mediterranean diet. 
We selected 27 vinegars and studied their antioxidant potential, total phenolic contents, sugar content and colour characteristics under the same conditions. Variability is reported in the levels of total phenolic contents ranging from 120 to 5680 mg/L depend on the colour and the origin of vinegar. The balsamic red vinegars had higher levels of phenolic compounds due to their longer fermentation process and also due to their origin. Red vinegars had higher phenolic contents as red wines  ADDIN EN.CITE <EndNote><Cite><Author>Woraratphoka</Author><Year>2007</Year><RecNum>51</RecNum><record><rec-number>51</rec-number><foreign-keys><key app="EN" db-id="ze50tdz2i2vsfjewxwaxeazof2zz2zdvpawv">51</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Jirayus Woraratphoka</author></authors></contributors><titles><title>Phenolic compounds and antioxidative properties of selected wines from the northeast of Thailand</title><secondary-title>Food Chemistry</secondary-title></titles><periodical><full-title>Food Chemistry</full-title></periodical><pages>1485-1490</pages><volume>104</volume><dates><year>2007</year></dates><urls></urls></record></Cite></EndNote>[18]. Phenolics are produced during the wine-making process. There are responsible for the colour, flavor and astringency of wine. During acetic fermentation of wine, the content of these wine phenolic compounds may decrease and may lead to different vinegar phenolic compound than those originally present in wine. 
Our results revealed also that the examined vinegars demonstrate high antioxidant potential, observed by BAP assay. The highest BAP values of all vinegars was predominant in the red balsamic vinegars, and this was quite expected, taking into account the concentrated grape must which balsamic vinegar is made of, the long ageing process in wooden casks which gives a very high concentration in phenols and other antioxidant substances. Natera et al., in a previous chemometric study of different kind of vinegars, according to their raw material, showed that the balsamic vinegars contains the highest amount of antioxidant substances such as gallic acid, tyrosol, cathechin and caffeic acid  ADDIN EN.CITE <EndNote><Cite><Author>Natera</Author><Year>2003</Year><RecNum>52</RecNum><record><rec-number>52</rec-number><foreign-keys><key app="EN" db-id="ze50tdz2i2vsfjewxwaxeazof2zz2zdvpawv">52</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Natera, R.</author><author>Castro, R.</author><author>de Valme Garcia-Moreno, M.</author><author>Hernandez, M. J.</author><author>Garcia-Barroso, C.</author></authors></contributors><auth-address>Analytical Chemistry Department, Faculty of Sciences, University of Cadiz, P.O. Box 40, E-11510, Puerto Real, Cadiz, Spain.</auth-address><titles><title>Chemometric studies of vinegars from different raw materials and processes of production</title><secondary-title>J Agric Food Chem</secondary-title></titles><periodical><full-title>J Agric Food Chem</full-title></periodical><pages>3345-51</pages><volume>51</volume><number>11</number><edition>2003/05/15</edition><keywords><keyword>Acetic Acid/*chemistry</keyword><keyword>Alcohols</keyword><keyword>Cereals</keyword><keyword>Food Handling/*methods</keyword><keyword>Fruit</keyword><keyword>Honey</keyword><keyword>Malus</keyword><keyword>Odors/analysis</keyword><keyword>Phenols/analysis</keyword><keyword>Time Factors</keyword><keyword>Volatilization</keyword><keyword>Wine</keyword><keyword>Wood</keyword></keywords><dates><year>2003</year><pub-dates><date>May 21</date></pub-dates></dates><isbn>0021-8561 (Print)&#xD;0021-8561 (Linking)</isbn><accession-num>12744665</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=12744665</url></related-urls></urls><electronic-resource-num>10.1021/jf021180u</electronic-resource-num><language>eng</language></record></Cite></EndNote>[19]. The high expected concentration of these antioxidant substances could explain high BAP values in our study and especially in balsamic red vinegars.
Davalos et al., have been demonstrated that the antioxidant capacity of wine vinegars is correlated to their polyphenolic content. The fruit grape products, wines and vinegars are rich in polyphenols  ADDIN EN.CITE  ADDIN EN.CITE.DATA [7, 20]. Previous study in sherry-derived vinegars showed that individual phenolic compounds which are well correlated to antioxidant activity were not necessarily those that occurred in high concentrations  ADDIN EN.CITE <EndNote><Cite><Author>Alonso A. M.</Author><Year>2004</Year><RecNum>54</RecNum><record><rec-number>54</rec-number><foreign-keys><key app="EN" db-id="ze50tdz2i2vsfjewxwaxeazof2zz2zdvpawv">54</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Alonso A. M., Castro R., Rodriguez M.C., Guillen D.A., Barroso C.G.</author></authors></contributors><titles><title>Study of the antioxidant power of brandies and vinegars derived from Sherry wines and corelation with their content in polyphenols.</title><secondary-title>Food Res Int</secondary-title></titles><periodical><full-title>Food Res Int</full-title></periodical><pages>715-721</pages><volume>37</volume><number>7</number><dates><year>2004</year></dates><urls></urls></record></Cite></EndNote>[21]. These findings seems to be confirmed by our results in which, although some colour indexes of vinegars decreased after 3 months storage, the antioxidant activity was not altered significantly. It is well known that the colour of wines is depended on their content in polyphenols, namely anthocyanins. Intense red and blue colour indicates high concentration of polyphenols, while yellow colour is indicative of low polyphenol content. In our tested vinegars, yellow colour was prominent in apple cider vinegars, ones with low polyphenol concentration.
In our study, we found also a positive correlation of sugar content and BAP value (fig. 1a) which could raise the question of a possible interference of sugars in our results. This does not seem to be the case since balsamic vinegars contain more sugars due to their production procedure but also the high phenolic content remains the stronger finding for the interpretation of their high BAP values. White balsamic vinegars also with high Brix value, but with low content of total phenols, showed much lower antioxidant activity.
The apple cider vinegars showed the lowest BAP values of all fruit vinegars and two of the lowest BAP values of all vinegar groups. Although apple cider vinegars have been commonly advertised, especially as lipotropic agents, their antioxidant activity is not considerable, probably due to the absence of anthocyanins and their low phenolic content, compared with red wine vinegars  ADDIN EN.CITE <EndNote><Cite><Author>van Velden</Author><Year>2002</Year><RecNum>56</RecNum><record><rec-number>56</rec-number><foreign-keys><key app="EN" db-id="ze50tdz2i2vsfjewxwaxeazof2zz2zdvpawv">56</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>van Velden, D. P.</author><author>Mansvelt, E. P.</author><author>Troup, G. J.</author></authors></contributors><auth-address>Department of Primary Medicine and Family Care, Stellenbosch University, Tygerberg, South Africa.</auth-address><titles><title>Red wines good, white wines bad?</title><secondary-title>Redox Rep</secondary-title></titles><periodical><full-title>Redox Rep</full-title></periodical><pages>315-6</pages><volume>7</volume><number>5</number><edition>2003/04/12</edition><keywords><keyword>Antioxidants/metabolism/*pharmacology</keyword><keyword>Cardiovascular System/drug effects</keyword><keyword>Female</keyword><keyword>Humans</keyword><keyword>Lipoproteins, LDL/metabolism</keyword><keyword>Male</keyword><keyword>Phenols/pharmacology</keyword><keyword>*Wine</keyword></keywords><dates><year>2002</year></dates><isbn>1351-0002 (Print)&#xD;1351-0002 (Linking)</isbn><accession-num>12688518</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=12688518</url></related-urls></urls><language>eng</language></record></Cite></EndNote>[22].
Keeping the vinegars tested at room temperature for a three months period showed that the antioxidant activity of each one was not altered significantly. We could suggest that, although the content of polyphenolic compounds in vinegars increases with the aging procedure in wood, such maturation of antioxidant capacity is taking place only in this producing period. Time of aging is able to change the phenolic profile of vinegars by increasing the polyphenolic index along time  ADDIN EN.CITE <EndNote><Cite><Author>Tesfaye</Author><Year>2002</Year><RecNum>44</RecNum><record><rec-number>44</rec-number><foreign-keys><key app="EN" db-id="ze50tdz2i2vsfjewxwaxeazof2zz2zdvpawv">44</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Tesfaye, W.</author><author>Morales, M. L.</author><author>Garcia-Parrilla, M. C.</author><author>Troncoso, A. M.</author></authors></contributors><auth-address>Area de Nutricion y Bromatologia, Facultad de Farmacia, Universidad de Sevilla, C/ P. Garcia Gonzalez s/n, E-41012 Sevilla, Spain.</auth-address><titles><title>Evolution of phenolic compounds during an experimental aging in wood of Sherry vinegar</title><secondary-title>J Agric Food Chem</secondary-title></titles><periodical><full-title>J Agric Food Chem</full-title></periodical><pages>7053-61</pages><volume>50</volume><number>24</number><edition>2002/11/14</edition><keywords><keyword>Acetic Acid/*chemistry</keyword><keyword>Aldehydes/analysis</keyword><keyword>Chemistry, Physical</keyword><keyword>Chromatography, High Pressure Liquid</keyword><keyword>Coumaric Acids/analysis</keyword><keyword>Discriminant Analysis</keyword><keyword>Ethanol/analysis</keyword><keyword>Furaldehyde/analysis</keyword><keyword>Hydrogen-Ion Concentration</keyword><keyword>Hydroxybenzoic Acids/analysis</keyword><keyword>Phenols/*analysis</keyword><keyword>Physicochemical Phenomena</keyword><keyword>Quercus</keyword><keyword>Time Factors</keyword><keyword>*Wine</keyword><keyword>*Wood</keyword></keywords><dates><year>2002</year><pub-dates><date>Nov 20</date></pub-dates></dates><isbn>0021-8561 (Print)&#xD;0021-8561 (Linking)</isbn><accession-num>12428959</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=12428959</url></related-urls></urls><electronic-resource-num>jf020602x [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>[11], but time of storage of the final product does not seem to influence the antioxidant activity, at least for the period and the conditions tested. In a previous study, the antioxidant capacity of wine industry by-products, remained stable even after 20 months storage in a dark and dry place at ambient temperature  ADDIN EN.CITE <EndNote><Cite><Author>Louli</Author><Year>2004</Year><RecNum>55</RecNum><record><rec-number>55</rec-number><foreign-keys><key app="EN" db-id="ze50tdz2i2vsfjewxwaxeazof2zz2zdvpawv">55</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Louli, V.</author><author>Ragoussis, N.</author><author>Magoulas, K.</author></authors></contributors><auth-address>Laboratory of Thermodynamics and Transport Phenomena, School of Chemical Engineering, National Technical University of Athens, 9 Heroon Polytechniou Str., Zografou Campus, 157 80 Athens, Greece. svlouli@chemeng.ntau.gr</auth-address><titles><title>Recovery of phenolic antioxidants from wine industry by-products</title><secondary-title>Bioresour Technol</secondary-title></titles><periodical><full-title>Bioresour Technol</full-title></periodical><pages>201-8</pages><volume>92</volume><number>2</number><edition>2003/12/25</edition><keywords><keyword>Acetates</keyword><keyword>Antioxidants/*isolation &amp; purification</keyword><keyword>Butylated Hydroxytoluene</keyword><keyword>Carbon Dioxide</keyword><keyword>Chromatography, High Pressure Liquid</keyword><keyword>Oils, Volatile</keyword><keyword>Phenols/*isolation &amp; purification</keyword><keyword>Pressure</keyword><keyword>*Rosmarinus</keyword><keyword>Vitis/*chemistry</keyword><keyword>Waste Disposal, Fluid/*methods</keyword><keyword>Wine/*analysis</keyword></keywords><dates><year>2004</year><pub-dates><date>Apr</date></pub-dates></dates><isbn>0960-8524 (Print)&#xD;0960-8524 (Linking)</isbn><accession-num>14693454</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=14693454</url></related-urls></urls><electronic-resource-num>S096085240300227X [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>[23].
In conclusion, our study indicates that vinegar has strong and stable antioxidant capacity when measured with BAP assay, the balsamic and red wine vinegars being the stronger than white and fruit vinegars, and this capacity is not altered significantly during a three month period under housekeeping conditions. To our knowledge, this is the first study that demonstrates the high antioxidant capacity of the most popular vinegars in the Greek market using the BAP assay, and highlights the quality of this seasoning in a Mediterranean diet.


Conflict of Interest

The authors declare that there are no conflicts of interest.
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20.	Davalos A, Bartolome B, Gomez-Cordoves C (2005)]qrt{�������		n	o	�	�	�	�	%
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D<EndNote><Cite><Author>Ahmed</Author><Year>2000</Year><RecNum>43</RecNum><record><rec-number>43</rec-number><foreign-keys><key app="EN" db-id="ze50tdz2i2vsfjewxwaxeazof2zz2zdvpawv">43</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Ahmed, R. S.</author><author>Seth, V.</author><author>Banerjee, B. D.</author></authors></contributors><auth-address>Department of Biochemistry, University College of Medical Sciences, Shahdara, Delhi 110095, India.</auth-address><titles><title>Influence of dietary ginger (Zingiber officinales Rosc) on antioxidant defense system in rat: comparison with ascorbic acid</title><secondary-title>Indian J Exp Biol</secondary-title></titles><periodical><full-title>Indian J Exp Biol</full-title></periodical><pages>604-6</pages><volume>38</volume><number>6</number><edition>2000/12/16</edition><keywords><keyword>Administration, Oral</keyword><keyword>Animal Feed</keyword><keyword>Animals</keyword><keyword>Antioxidants/*pharmacology</keyword><keyword>Ascorbic Acid/*pharmacology</keyword><keyword>Catalase/blood</keyword><keyword>Ginger/*chemistry</keyword><keyword>Glutathione/blood</keyword><keyword>Glutathione Peroxidase/blood</keyword><keyword>Glutathione Transferase/blood</keyword><keyword>Lipid Peroxidation/*drug effects</keyword><keyword>Male</keyword><keyword>*Plants, Medicinal</keyword><keyword>Powders</keyword><keyword>Rats</keyword><keyword>Rats, Wistar</keyword><keyword>Superoxide Dismutase/blood</keyword><keyword>Thiobarbituric Acid Reactive Substances/analysis</keyword></keywords><dates><year>2000</year><pub-dates><date>Jun</date></pub-dates></dates><isbn>0019-5189 (Print)&#xD;0019-5189 (Linking)</isbn><accession-num>11116533</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=11116533</url></related-urls></urls><language>eng</language></record></Cite><Cite><Author>Davalos A</Author><Year>2004</Year><RecNum>65</RecNum><record><rec-number>65</rec-number><foreign-keys><key app="EN" db-id="ze50tdz2i2vsfjewxwaxeazof2zz2zdvpawv">65</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Davalos A, Bartolome B, Gomez-Cordoves C.</author></authors></contributors><titles><title>Antioxidant properties of commercial grape juices and vinegars</title><secondary-title>Food Chemistry</secondary-title></titles><periodical><full-title>Food Chemistry</full-title></periodical><pages>325-330</pages><volume>93</volume><section>325</section><dates><year>2004</year></dates><urls></urls></record></Cite></EndNote>��@@�@U~:������CJ_HaJmHsHtHJ@JU~:
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