Elemental Profile and 207 Pb / 206 Pb , 208 Pb / 206 Pb , 204 Pb / 206 Pb , 87 Sr / 86 Sr Isotope Ratio as Fingerprints for Geographical Traceability of Romanian Wines

Geographical wine traceability is an important topic in the context of wine authentification. Therefore, many researchers have addressed this subject by developing different methodologies based on multivariate analysis of organic and inorganic parameters and also by isotopic signature. The goal of this research was to assess the potential of elemental composition and isotopic signature of lead (207Pb/206Pb, 208Pb/206Pb and 204Pb/206Pb) and strontium (87Sr/86Sr) of wines from three Romanian vineyards, in order to highlight reliable markers for wine geographical origin. The ICP-MS method was used for the concentration determination for 30 elements (Ag, Al, As, Ba, Be, Bi, Ca, Cd, Co, Cr, Cs, Cu, Fe, Ga, In, K, Li, Mg, Mn, Na, Ni, Pb, Rb, Se, Sr, Tl, V, U, Zn, Hg). In this study 10 wines (2 red and 8 white) obtained from ‘Merlot’, ‘Fetească neagră’, ‘Fetească albă’, ‘Fetească regală’, ‘Băbească gri’, ‘Șarba’, ‘Aligoté’, ‘Sauvignon blanc’, ‘Muscat Ottonel’, ‘Italian Riesling’ cultivars were investigated. The wine samples were obtained from micro-wine production under conditions of 2014, 2015, 2016 from Dealu Bujorului, Murfatlar and Ștefănești-Argeș vineyards. The high level of K (148.66±5.41-633.74±4.13 mg/L), Mg (88.23±0.84-131.66±3.42 mg/L), Ca (49.84±1.22-89.18±2.34) and Fe were observed in the wine samples analysed. Heavy metals like Hg, Pb, As and Cd (10.2-315 μg/L) were found below acceptable limits. Concentration of Na (1 mg/L), Cu (1 mg/L), As (0.2 mg/L), Cd (0.01 mg/L), Zn (5 mg/L) and Pb (0.15 mg/L) metals in analysed wine samples were under Maximum Permissible Limits (MPL), respectively as published by the Organization of Vine and Wine. The variation of the 207Pb/206Pb, 208Pb/206Pb, 204Pb/206Pb and 87Sr/86Sr ratio and K/Rb, Ca/Sr of the investigated wine clearly demonstrated that these variables are suitable traces for wine geographical origin determination. The proposed methodology allowed a 100% successful classification of wines according to the region of provenance.


Introduction
The traceability of foods has become a priority among consumers, driven by the increasing demand for food quality and food safety (Martins et al., 2013).In the traceability system, the discrimination of the geographical provenance of food products is essential to verify the claims of origin of the declared-on labels and to prevent unsafe products from reaching the consumers (Barbaste et al., 2002).The geographical origin assessment of wine is of particular interest, being one of the most important factors that determine its commercial values (Petrini et al., 2015).Consumers are attributing lately more values regarding the certification of food products, their origins and authenticity (Vinciguerra et al., 2015).
To establish the geographical origin of wines is a major concern issue for many countries around the world, in order to protect quality products in the case of false statements between the wine composition and the geochemistry of provenance soil (Vorster et al., 2010).The elemental profile of wine may be affected by several factors such as agricultural practices, environmental contamination, climatic change and also the winemaking process, which affects the wine elements content (Almeida and Vasconcelos, 2003).
The control of the geographical origin of wine based on its chemical composition is one of the most challenging issues in relation to wines authenticity.In the last decade, many efforts have been made for identify potential markers and develop reliable analytical methods to determine the wines authenticity (Martins et al., 2013).The application of methods using stable isotopes of light elements (H, C, O, S and N) have been started since the beginning of the 1950, providing information on climate, distance from the sea, latitude, altitude and technological practices (Morgun et al., 2008;Loftus et al., 2016).
More recently, the study of isotopic ratios of heavy metals such Pb and Sr came into use in this field application, providing additional information on the geographical origin, since plants inherit the isotopic signature of these elements from the geological and pedological environmental (Horn et al., 1993;Barbaste, 2001;Rummel et al., 2010;Martins et al., 2013).
Strontium is found in nature as three abundant isotopes: 86 Sr (9.75-9.99%), 87Sr (6.94-7.14%), 88Sr (82.29-82.77%)and 84 Sr (0.55-0.58%) as less abundant isotope (Berglund and Wieser, 2011;Martins et al., 2013).The 87 Sr is radiogenic and therefore the 87 Sr content increases with time due to radioactive decay of 87 Rb (Geana et al., 2016).Since the content of 87 Sr in soil varies with geological age and geographical location, the 87 Sr/ 86 Sr isotopic ratio can be used as a tracer for determining the geographical origin of wine (Vorster et al., 2010).Many studies have shown that there is a significant correlation between isotopic ratio 87 Sr/ 86 Sr from wines and soils of origin (Di Paola-Naranjo et al., 2011;Durante et al., 2013), thus representing the starting premise for geographical origin determination.The isotopic ration 87 Sr/ 86 Sr is a well-established tool for dating and tracing the origin of rocks and also minerals (Capo et al., 1998) with special interest of wine traceability.Several studies on its use for wine geographical origin assessment can be found in literature (Horn et al., 1993;Almeida and Vasconcelos, 2004;Durante et al., 2013;Catarino et al., 2016).Recently, within a research program regarding strategies for wine fingerprinting, the authors confirmed 87 Sr/ 86 Sr as a viable tool for traceability of Romanian PDO and GI wines (Geana et al., 2016), Portuguese PDO, where soils were developed on different geological formations (Martins et al., 2014;Catarino et al., 2016).The use of this parameter as a marker of wine geographical origin is based on the assumption that a relation between soil, plants and wine exists.Therefore, 87 Sr/ 86 Sr ratio should not be significantly modified during wine processing.Aiming to use 87 Sr/ 86 Sr for traceability and authentication of wine, it is mandatory to clarify the impacts of anthropogenic factors and technological processes on this isotopic ration (Catarino et al., 2016).
Determination of the elemental composition of wines is useful for many reasons.Firstly, the concentration of elements in wine is useful information to vine grower and oenologists for controlling the process of obtaining high and quality wines (Pohl, 2007).Secondly, the elemental composition could be used as a wine fingerprint and represents one of the criteria for evaluating the authenticity of wine (Grindlay et al., 2008).
The minerals in wine originate from the capacity of the vine to take elements from soil (geographical region), the climatic factors such as heavy rains, environmental conditions (pollution), and also agricultural applications pesticides and fertilizers (Şen and Tokatli, 2014).The mineral content in white and red wines from the same region can differ due to the impact of the vinification process on the elemental composition, such as the maceration step in red winemaking, where the juice is in longer contact whit the skins and flesh of the grapes (Coetzee et al., 2005).
Based on these factors, the elements in wine can be classified into two groups: endogenous and exogenous (Grindlay et al., 2011).The most abundant are endogenous elements (Ca, Mg, Na, K, Zn, and P) that are related to the grape variety and maturity, type of soil in the vineyard, and ecoclimatic conditions (Ðurđić et al., 2017).The exogenous elemental content (Al, Cd, Cr, Cu, Fe, and Zn) depends on external impurities during the growth of grapes and vinicultural and winemaking practices (Pohl, 2007;Ivanova-Petropulos et al., 2013;Versari et al., 2014).Anthropogenic factors, such as application of fungicides, pesticides and fertilisers during the growing season, can lead to an increase in Cd, Cu, Mn, As, and Zn in wine.The presence of Pb in wines can originate from sources like traffic, fertilisers, vessels, and pumps used during vinification process (Almeida and Vasconcelos, 1999).Recent data indicate that some beverages, including wine (red and also white), contribute to the total dietary intake of certain trace elements (B, Ba, Co, Mn, Ni, Rb, Sb, Tl, and V) (Grindlay et al., 2011;Tariba et al., 2011;Ivanova-Petropulos et al., 2016) and iron as well (Ðurđić et al., 2017).
The most used chemical parameters for discriminating the geographical origin of wine are the stable isotope ration of oxygen, carbon and hydrogen (Dordevic et al., 2012;Raco et al., 2015;Geana et al., 2016) and also the elemental composition (Fabani et al., 2010;Selih et al., 2014) including rare elements (Gonzálvez et al., 2009) and 87 Sr/ 86 Sr isotope ratio (Durante et al., 2013;Marchionni et al., 2013;Geana et al., 2016) or a combination of them (Dutra et al., 2011).The ability to differentiate wines by regions through their elemental patterns suggests that elements are mainly regulated by their migration from rock to soil and from vineyard soil to grapes.Elements are taken up by the roots passing to the grapes in the same isotopic proportions as they occur in the soil (Almeida and Vasconcelos, 2003).
Generally, the elements which are selected as tracers for geographic origin of wines must have a strong correlation 56.21%) and 204 Pb (1.04-1.65%)as less abundant isotope (Rossman and Taylor, 1998).Their abundance extensively varies because of different decay pathways from 238 U, 235 U and 232 Th to 206 Pb, 207 Pb, 208 Pb respectively (Faure, 1986).The Pb isotope of ore deposits and anthropogenic sources has their distinct isotopic ratios or signatures (Cheng and Hu, 2010).The Pb isotope ratio did not change in industrial or environmental processing and retained its characteristic ratio from source ore (Ault et al., 1970).The Pb isotope ratio can be used to identify the sources and transport pathways of Pb in pollution studies (Hu et al., 2015).Previous works have shows that the use of lead isotopes is a good tool in order to know the origin of Pb and to identify the types of Pb according to anthropogenic or geogenic origin (Ettler et al., 2004;Li et al., 2011;Álvarez-Iglesias et al., 2012); this is because the Pb emitted into the atmosphere maintains a characteristic isotopic composition (signature) and does not change during the physical/ physicochemical processes associated with smelting, refining or manufacturing (Flegal and Smith, 1995).The Pb from anthropic sources has less radiogenic ratios than from geogenic sources (1.21-1.33, 206Pb/ 207 Pb).Therefore, the sources of Pb pollution heave specific ration that allow them to be differentiated.Many studies have shown that there is a significant correlation between isotope ration 206 Pb/ 207 Pb, 208 Pb/ 206 Pb from wines and soil of origin (Avram et al., 2014;Kristensen et al., 2016;Dehelean and Voica, 2012;Almeida et al., 2016), this representing the starting premise for geographical origin determination.The lead isotope ratio depends on factors such as thorium and uranium content in the soil, weathering processes and original rock age, which provide a fingerprint used for different forensic and archeological purposes (Komárek et al., 2008;Roux et al., 2004;Dreyfus et al., 2007).Lead isotopic analysis of wines from Bordeaux, France showed that lead from wines changed over time to reflect the dominant source of atmospheric lead pollution in southern France (Médina et al., 2000).
The premise from which we started this research is that the elemental profile of wines reflects the chemical composition of vineyard soil (Coetzee et al., 2005;van der Linde et al., 2010).The aim of the present research is to determine the elemental composition and 207 Pb/ 206 Pb, 208 Pb/ 206 Pb, 204 Pb/ 206 Pb and 87 Sr/ 86 Sr isotopic ratio for red and white wines from three different Romanian winemaking regions, in order to highlight reliable markers for wine geographical origin.

Sample collection and microvinification process
The samples used in this experiment were obtained from the wines produced from 'Merlot', 'Fetească neagră', 'Fetească albă', 'Fetească regală', 'Băbească gri', 'Șarba', 'Aligoté', 'Sauvignon blanc', 'Muscat Ottonel', 'Italian Riesling' under the conditions of 2014, 2015 and 2016 year, from Dealu Bujorului, Murfatlar and Ștefănești-Argeș vineyard.The wine samples resulted from micro-wine production.Micro-vine production it was done according to the methodology described by Bora et al. (2016).All wines were providing by the wineries as finished wines in 750 mL glass bottles with cork stoppers and were stored at 3-4 °C before analysis.One bottle was used for each sample, and three replicates were taken.All vines were planted since 1979, and the vine plantation was organized with 2.2 × 1 m distance between rows and plants.Vines were pruned according to the Guyot system and were grown on speliers.

Reagents and solutions
Thirty elements (Ag, Al, As, Ba, Be, Bi, Ca, Cd, Co, Cr, Cs, Cu, Fe, Ga, In, K, Li, Mg, Mn, Na, Ni, Pb, Rb, Se, Sr, Tl, V, U, Zn and Hg) were determined in order to assess their ability to discriminate wines by geographical origin.The analysis was made using multielement analysis and ICP-MS technique, after an appropriate dilution, using external standard calibration method.Each sample was analyzed in duplicate and each analysis was prepared from consisted of three replicates.The calibration was performed using XXICertiPUR multielement standard, and from individual standard solution of Hg.The working standards and the control sample were prepared daily from the intermediate standards that were prepared from the stock solution.The intermediate solutions stored in polyethylene bottles and glassware was cleaned by soaking in 10% v/v HNO3 for 24 hours and rinsing at least ten rimes with ultrapure water (Milli-Q Integral ultrapure water-Type 1).The accuracy of the methods was evaluated by replicate analyses of known concentration samples (between 10 µL to 10 mL concentrations) and the obtained values ranged between 0.8-13.1 percent, depending on the element.The global recovery for each element was estimated and the obtained values were between 84.6-100.9%(Geana et al., 2016).
For quality control purpose, blanks and triplicates samples (n = 3) were analyzed during the procedure.The variation coefficient was under 5% and detection limits (ppb) were determined by the calibration curve method.
Limit of detection (LoD) and Limit of quantification (LoQ) limits were calculated according to the next mathematical formulas: LoD = 3SD/s and LoQ = 10 SD/s (SD = estimation of the standard deviation of the regression line; s = slope of the calibration curve).
To verify the achieved accuracy and precision, ten NIST-SRM 987 and NIST-SRM 982 analysis results were pooled together with the calculated relative standard deviation presented in Table 2. Based on the obtained results, it was verified that, applying quadrupole ICP-MS, relative standard deviation and reproducibility of approximately 0.5% for 87 Sr/ 86 Sr, 206 Pb/ 207 Pb and 208 Pb/ 206 Pb are feasible.The results were in agreement with those reported by (Ketterer et al., 1991;Barbaste et al., 2001;Almeida et al., 2016;Geana et al., 2016).

Sample preparation for determination of metals and isotope ratio from wine using ICP-MS
For the determination of metals from wine samples were used an amount of 0.5 mL wine and adjust 8 mL (7 mL HNO3 65%+1 mL H2O2), after 15-30 minutes the mineralization was performed using a microwave system Milestone START D Microwave Digestion System set in three steps: step I (time 10 min., temperature 200 °C), step II (time 15 min., temperature 200 °C) and step III (time 60 min., ventilation -temperature 35 °C).

Instrumentation
The determination of metals was performed on mass spectrometer with inductively coupled plasma (ICP-MS) iCAP Q Thermo scientific model, based polyatomic species before they reach the quadrupole mass spectrometer, using a PFA micro flow concentric nebulizer.The argon used was of 99.99% purity (Messer, Austria).The instrument was daily optimized to give maximum sensitivity for M + ions and the double ionization and oxides monitored by the means of the ratio between Ba 2+ /Ba + and Ce 2+ /CeO + , respectively, these always being less than 2%.The experimental conditions were: argon flow on nebulizer (0.84 L/min.), auxiliary gas flow 0.80 L/min., argon flow in plasma 15 L/min., lens voltage 7.31 V; RF power in plasma 1100 W, spray chamber temperature (2.51±1.00o C).Accuracy was calculated for the elements taken into consideration (0.5-5.0%).

Statistical analysis
The statistical interpretation of the results was performed using the Duncan test, SPSS Version 24 (SPSS Inc., Chicago, IL., USA).The statistical processing of the results was primarily performed in order to calculate the following statistical parameters: average and standard deviation.This data was interpreted with the analysis of variance (ANOVA) and the average separation was performed with the DUNCAN test at p ≤ 0.005.Linear discriminant analysis (LDA) was performed in order to separate the wines by region and to indentify the markers with a significant discrimination value (variables with Wilk's lambda near zero, p values <0.005 and higher F coefficients).Linear discriminant analysis (LDA) was performed using Microsoft Excel 2016 and XLSTAT Addinsoft version 15.5.03.3707.

Wine mineral content
These elements are present in grapes as results of their accumulation in the vine plant through the root from the soil, or they could originate from the agents used in protecting the vine from diseases.During the maceration, extracted elements are absorbed at the cell membrane of yeast, and afterward, their declines as a result of precipitation together with the yeast cell or precipitation in complexation reactions.
The elemental contents of the investigated wines according to the geographical origin are present in Appendix 1, 2, 3 as mean and standard deviation.As expected potassium was the most abundant element in all investigated red and white wine samples since this element is essential for the growth and development of plants and is often a component of fertiliser (Rodrigues et al., 2011).According to our results, the K concentration was higher in red compared to white wines.The measured values ranged between 491.12±4.49to 633.74±4.13mg/L for red wines and from 148.66±5.41 to 327.64±9.00mg/L for white wines (Appendix 3).
These results agree with values reported in the literature (Iglesias et al., 2007 - The same as Mg concentration, the average content of Na differed among the regions and decreased in the order Dealu Bujorului > Ștefănești-Argeș > Murfatlar.The highest concentration of Na (55.59±1.11mg/L Șarba (2014)) was found in wines Dealu Bujorului.The sodium concentration was higher in red wines (mean 44.74 mg/L, values between 36.89±0.30 to 51.62±0.64)than in white wines (mean 42.08 mg/L, values between 31.35±1.25 to 55.59±1.11).These results are in agree with Ražić and Onjia (2010) where they obtained the similar concentration of sodium in wine from North Serbia (33±11 mg/L).The Na content in our study are similar with the results published on Serbian (Ražić and Onjia, 2010 -average values of 29.65 mg/L Na), Czech (Kment et al., 2005 average values of 14.7 mg/L Na) and Spanish (Iglesias et al., 2007 -average values of 37.19 mg/L Na) wines.The higher content of sodium in Dealu Bujorului and Murfatlar wines has been attributed to the vicinity of the Black Sea.
The content of trace elements (In, Sr, Ni, Rb, Se, Tl, U, Zn, Ag, Al, Be, Bi, Ba, Cr, Cs and Ga) found in Romanian wines agreed with literature data (Kment et al., 2005;Filket et al., 2011;Ivanova-Petropulos et al., 2013;Geana et al., 2013).Regarding Ni, Rb, Se, Tl, U, Ag, Cr and Cs the highest concentration were obtained at red wines, and in the case of In, Sr, Zn, Al, Be, Bi and Ba the highest concentration were obtained at white wines.The results indicated that Romanian red wines are moderately rich in In, Sr, Ni, Rb, Se, Tl, U, Zn, Ag, Al, Be, Bi, Ba, Cr, Cs and Ga while white wines are moderately rich In, Sr, Zn, Al, Be, Bi and Ba (Appendix 2-3).
In general Area, Variety, Years, Area × Variety, Area × Years, Variety × Years and Area × Variety × Years factors influenced accumulation of Ca, Mg, Fe, V, Ag, Al, Bi, Ba, Cr, Cs, In, Sr, Ni, Rb, Se, Tl, U, Zn, Hg and Pb.
Concerning the influence factors on the accumulation of metals in wines, it can be seen that in case of Ca, Mg, Fe, V, Ag, Al, Bi, Ba, Cr, Cs, In, Sr, Ni, Rb, Se, Tl, U, Zn, Hg and Pb was significantly influenced by Area, Variety, Years, Area × Variety, Area × Years, Variety × Years and Area x Variety x Years factors and in case of K, Li, Cu, Na, Mn, Co, As Be, Cd and Ga was significantly influenced distinctive by the factors taken into consideration.
Concentration of Na (1 mg/L), Cu (1 mg/L), As (0.2 mg/L), Cd (0.01 mg/L), Zn (5 mg/L) and Pb ( 0 Pb, 207 Pb and 208 Pb (radiogenic) originated from the genesis of the substrate and does not varies with geological ages.The original composition of soil samples retains its chemical composition from the geographical area it belongs to (Shirahata et al., 1980;Gulson et al., 1981;Elbaz-Poulichet et al., 1984).This property is useful in order to identify of the source of lead in a subjected wine sample provided that the measurements of the isotope ratio is precise and accurate.
The Lower87 Sr/ 86 Sr isotope ratio of wine from Murfatlar area is due to influence of saline aerosols, with a 87 Sr/ 86 Sr isotope ratio of about 0.70917 (Rodrigues et al., 2011;Geana et al., 2016).
The ratio of different metals (K/Rb and Ca/Sr) are important due to the fact that the K/Rb ratio can differ significantly between different roks and soil and the Ca/Sr ration is used as a chemical tracer in the biochemistry and hydro-geochemistry studies (Di Paola-Naranjo et al., 2011;Geana et al., 2016).Concerning the element ratio, wine from the Ștefănești-Argeș area (898.89±4.21)have higher K/Rb ration, those from the Dealu Bujorului (675.44±4.53)shows higher K/Rb ratio.Wine from the Murfatlar area shows lower Ca/Sr ratio (21.09±1.08)and wine from Dealu Bujorului shows lower Ca/Sr ratio (14.83±0.98)(Table 3).
Variation of the Based on the elemental contents and metal ratio (K/Rb and Ca/Sr) data, the cross-validation technique provided a 100 % percentage of predicted membership according to the wine geographic origin (F1 = 65.28% and F2 = 34.72%)(Fig. 1).The linear correction revealed acceptable scores for the two defined discriminant factors (F1 and F2).A significant differentiation of wines according to the geographic origin was carried out for wines, which demonstrates the importance of elemental profile for the geographical traceability of wines.
processes.Thus, Ca and other elements like Cu, K, which are affected by exogenous factors, were not considered for wines geographical discrimination (Geana et al., 2016).
Multivariate chemometric method was applied for the differentiation of wines intro groups on the basis of their geographic origin.Stepwise linear discriminant analysis (LDA) was used to identify significant tracers for classification to the geographical discrimination of the wines samples.By cross-validation, we established the optimal number of parameters required to obtain a robust model.Sr, isotope ratio for wine geographical discrimination, a 100% percentage of predicted membership according to the wine geographic origin was obtained (F1 = 78.51% and F2 = 21.49%) (Fig. 2).

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Wines were grouped in there distinct groups corresponding to regions of provenance.Moreover, the first discriminant function separated mainly the wines from Murfatlar and Dealu Bujorului from Ștefănești-Argeș area and the second one discriminated mainly the wines from Murfatlar from Dealu Bujorului and Ștefănești-Argeș area.Pb/ 206 Pb, 87 Sr/ 86 Sr were identified as the most significant for geographic differentiation of the wine (Fig. 3).
Relevant results for wine geographical classification were achieved, in both cases, with or without the 207 Pb/ 206 Pb, 208 Pb/ 206 Pb, 204 Pb/ 206 Pb, 87 Sr/ 86 Sr isotope ratio variable, which means that lead and strontium isotope analysis is not imperative for discrimination of wines according to their geographic origin, elemental profile and certain metal ratios being sufficient for this purpose.The technique of crossvalidation was applied during the set validation and the proposed model appears to be a promising chemometric approach for precise classification of wines according to their geographical origin.Thus, in both cases, the geographical regions were correctly classified with percentage between 52 and 71 %.

Conclusions
In this study the characterisation of Romanian wines according to their elemental composition was performed.Potassium, calcium and magnesium were the most abundant elements in all investigated wine samples.Concentration of Na (1 mg/L), Cu (1 mg/L), As (0.2 mg/L), Cd (0.01 mg/L), Zn (5 mg/L) and Pb (0.15 mg/L) metals in analysed wine samples were under Maximum Permissible Limits (MPL), respectively as published by the Organization of Vine and Wine.Based on the elemental profile, the K/Rb and Ca/Sr metal ratio and 207 Pb/ 206

Fig. 1 .
Fig. 1.Differentiation of wines according to geographic origin based on element contents and K/Rb and Ca/Sr ratios Fig. 2. Differentiation of wines according to geographic origin based on element contents and K/Rb, Ca/Sr ratios and 207 Pb/ 206 Pb, 208 Pb/ 206 Pb, 204 Pb/ 206 Pb, 87 Sr/ 86 Sr
(Coetzee et al., 2014061 mg/L;Álvarez  et al., 2012-average values of 865.30 mg/L).Magnesium and calcium were the second abundant elements in our study.The magnesium concentration was higher in red wines (mean 100.67 mg/L, values between 99.91±1.35 to 129.28±5.64)than in white wine (mean 93.76 mg/L, values between 88.23±0.84 to 131.66±3.42),whilecalciumconcentrationweresimilar in both types of wines (62.56 mg/L and 69.60 mg/L for red and white wines, respectively).The values obtained for the Mg and Ca contents in our selected wines were in good agreement with the results for Macedonian (Ivanova-Petropulos et al., 2013average values of 83.5 mg/L Ca and 98.20 mg/L Mg), Serbian(Ražić and Onjia, 2010-average values of 37 mg/L Ca and 95.73 mg/L Mg), Croatian(Vrček et al., 2011average values of 65.90 mg/L Ca and 68.70 mg/L Mg) and also Czech wines(Kment et al., 2005-average values of 108.00 mg/L Ca and 75.40 mg/L Mg).On the other hand, our Ca and Mg contents were significantly higher than published data for wines from Argentina(Lara et al., 2005average values of 12.50 mg/L Ca) and Belgium(Coetzee et al., 2014-average values of 6.73 mg/L Ca and 12.05 mg/L Mg).