Effects of reduced levels of sulfite in wine production using mixtures with lysozyme and dimethyl dicarbonate on levels of volatile and biogenic amines

Sulphur dioxide (SO₂) is an important preservative for wine, but its presence in foods can cause allergies and this has given impetus to the research for alternatives. The aim of this study was to reduce levels of sulfite in wine production using mixtures with lysozyme and dimethyl dicarbonate and examine the influence on levels of volatile and biogenic amines. To do so, vinifications were carried out using lysozyme, dimethyl dicarbonate (DMDC) and mixtures of these with SO₂ in different concentrations (25 and 50 mg l^?1). Results were compared with a control vinification with only SO₂ (50 mg l^?1). Mixing low concentrations of SO₂ with lysozyme and DMDC reduced the concentration of biogenic amines (histamine, tyramine, putrescine, cadaverine, phenylethylamine + spermidine and spermine). In general, the total concentration of volatile amines (dimethylamine, isopropylamine, isobutylamine, pyrrolidine, ethylamine, diethylamine, amylamine and hexylamine) was higher in the sample fermented only with SO₂. The concentrations of amines with secondary amino groups (dimethylamine, diethylamine, pyrrolidine) were higher in the sample only fermented with SO₂ than those fermented with DMDC and lysozyme or with a mixture of preservatives. When SO₂ was the only preservative in wine, total amine concentration (biogenic and volatile amines) was higher than for the rest of the treatments. Lysozyme by itself, and lysozyme mixed with SO₂, both reduced the formation of biogenic amines but given the antioxidant activity of SO₂ the use of the preservative mixture seems more advisable.     

Comparison of amino acids,biogenic amines and ammonium ion of wines made of different types of fruits

Amino acids, biogenic amines and ammonium ion are the primary nitrogen sources in fruit wines. Compared to grape wines, our knowledge to their profile in wines made of other fruits is very lack. Fourteen wines made of nine kinds of fruits together with seven grape wines were selected to compare their nitrogen component profile. White grape wines possessed the highest level of total amino acids, followed by red grape wines and then the wines made of the other fruits. Red grape wines contained the highest level of biogenic amines, followed by white grape wines and the wines made of the other fruits. Ammonium ion was also found to be higher in the wines made of grapes. Principal component analysis revealed that the obvious separations existed among the wines made from different types of fruits, indicating that these nitrogen compounds could be served as the indicators to differentiate different types of fruit wines.     

Pre-fermentative replacement of sulphur dioxide by lysozyme and oenological tannins: Effect on the formation and evolution of volatile compounds during the bottle storage of white wines

In this work, the effects on the volatile profile of the pre-fermentative substitution of SO₂ with lysozyme and oenological tannins were studied in white wines. At the same time, in order to understand the changes of volatile compounds in SO₂-free wines, the evolution of volatiles was evaluated over 1 year of storage in bottles. For this purpose, a number of laboratory scale fermentations of SauvignonBlanc musts were carried out and the effects of three variables (SO₂, lysozyme and oenological tannins) were investigated by means of GC–MS analysis. Results showed that the replacement of SO₂ with lysozyme and oenological tannins influenced the volatile composition of wines at the end of the alcoholic fermentation. Wines fermented with SO₂ showed higher total alcohol amounts, while the presence of oenological tannins augmented the level of esters. The presence of SO₂ influenced also the alcohols and esters profiles of wines during bottle storage. Moreover, the presence of oenological tannins displayed a positive role in maintaining the amounts of esters over certain levels in wine stored for 1 year, likely due to their oxygen scavenging ability. By contrast, acids were less affected by the investigated adjuvants both at the end of the alcoholic fermentation and during the storage time.     

Improvement of wine aromatic quality using mixtures of lysozyme and dimethyl dicarbonate,with low SO2 concentration

The use of sulphur dioxide (SO₂) in the treatment of foodstuffs presents some problems as it could lead to pseudo-allergies in some people. The aim of this research work was to study the addition of different preservative mixtures and their influence on the concentration of volatile compounds and sensorial quality in wine. To do so, vinifications were carried out using Garnacha must to which lysozyme, dimethyl dicarbonate (DMDC) and mixtures of these with SO₂ were added at different doses (25 and 50 mg l^?1). The results were compared with a control sample to which only SO₂ had been added (50 mg l^?1). In general, mixtures of SO₂ with lysozyme and DMDC favoured the formation of volatile compounds in the wines. Wines obtained from the mixtures of lysozyme and DMDC with 25 mg l^?1 of SO₂ had better sensorial quality than the wines obtained with 50 mg l^?1 as the only preservative used.     

Replacement of sulfur dioxide by lysozyme and oenological tannins during fermentation: influence on volatile composition of white wines

BACKGROUND: In recent years the use of sulfur dioxide, a commonly used additive in winemaking, has been questioned because of its toxic effects on human health. Studies have been conducted to find alternatives that can effectively substitute for this additive in all its various technological functions. In previous work, lysozyme and oenological tannins were found as possible substitutes in controlling bacterial undesirable fermentations and phenolic oxidation. However, data on the volatile composition of wines obtained by that protocol are lacking. In this work, the effects on volatile composition of white wines by the substitution of SO₂ during fermentation with lysozyme and tannin were studied. At the same time, the technological performance of two strains of yeast that produce low amounts of SO₂ were evaluated. RESULTS: The results showed that both SO₂ and lysozyme prevented the development of undesirable bacterial fermentations. The study of volatile compounds shows differences in the alcohol, acid and ester contents among the final products: wines fermented with strain 1042 and lysozyme had higher total alcohol concentration, while the addition of SO₂ promoted higher production of 3‐methyl‐1‐butanol, 3‐methylthio‐1‐propanol, phenylethyl alcohol and 4‐hydroxy‐benzenethanol. Esters, as a total, were influenced by the different strain and tannins addition, while amounts of medium‐chain fatty acid ethyl esters and their corresponding fatty acids were found in higher amounts in wines coming from fermentations with lysozyme. The sensory analysis revealed a preference for wines to which lysozyme and tannins had been added. CONCLUSION: The data suggest that the addition of lysozyme and oenological tannins during alcoholic fermentation could represent a promising alternative to the use of SO₂ and for the production of wines with reduced content of SO₂. The composition of the volatiles in the final wines was affected by the different vinification protocols (mainly with regards to alcohols and ethyl esters). Copyright © 2009 Society of Chemical Industry     

Amino acids and biogenic amines in red varietal wines: the role of grape variety, malolactic fermentation and vintage

This work studies for the first time the amino acid and biogenic amine composition of Rioja red wines made with the red minority varieties Vitis vinífera cv. Monastel and Maturana Tinta de Navarrete, using Tempranillo as a reference variety. The role of malolactic fermentation and vintage on these compounds was also analysed, and discriminate analyses were applied to achieve a possible differentiation of the wines. Amino acid composition allowed a differentiation of wines according to grape variety. Monastel was characterised by the highest value in β-alanine and Maturana Tinta de Navarrete by its highest value in OH-proline. However, biogenic amines were no able to classify varietal wines. The malolactic fermentation had significant changes on the amino acid and biogenic amine content, and allowed distinguishing wines that underwent this process from wines without malolactic fermentation. No correlation was found between total amino acids and total biogenic amines after malolactic fermentation, suggesting that a higher initial concentration of amino acids in the medium did no affect the concentration of biogenic amines after malolactic fermentation. Vintage influenced the amino acid and biogenic amine pattern, obtaining a clear separation of wines by vintages. Monastel and Maturana Tinta de Navarrete wines showed a minor varietal character and were more influenced by the climatic conditions of each vintage than Tempranillo wines. All the wines showed histamine levels below the human physiological threshold and implemented regulations.     

Implications of nitrogen nutrition for grapes, fermentation and wine

This review discusses the impacts of nitrogen addition in the vineyard and winery, and establishes the effects that nitrogen has on grape berry and wine composition and the sensory attributes of wine. Nitrogen is the most abundant soil‐derived macronutrient in a grapevine, and plays a major role in many of the biological functions and processes of both grapevine and fermentative microorganisms. Manipulation of grapevine nitrogen nutrition has the potential to influence quality components in the grape and, ultimately, the wine. In addition, fermentation kinetics and formation of flavour‐active metabolites are also affected by the nitrogen status of the must, which can be further manipulated by addition of nitrogen in the winery. The only consistent effect of nitrogen application in the vineyard on grape berry quality components is an increase in the concentration of the major nitrogenous compounds, such as total nitrogen, total amino acids, arginine, proline and ammonium, and consequently yeast‐assimilable nitrogen (YAN). Both the form and amount of YAN have significant implications for wine quality. Low must YAN leads to low yeast populations and poor fermentation vigour, increased risk of sluggish/stuck/slow fermentations, increased production of undesirable thiols (e.g. hydrogen sulfide) and higher alcohols, and low production of esters and long chain volatile fatty acids. High must YAN leads to increased biomass and higher maximum heat output due to greater fermentation vigour, and increased formation of ethyl acetate, acetic acid and volatile acidity. Increased concentrations of haze‐causing proteins, urea and ethyl carbamate and biogenic amines are also associated with high YAN musts. The risk of microbial instability, potential taint from Botrytis‐infected fruit and possibly atypical ageing character is also increased. Intermediate must YAN favours the best balance between desirable and undesirable chemical and sensory wine attributes. ‘Macro tuning’, of berry nitrogen status can be achieved in the vineyard, given genetic constraints, but the final ‘micro tuning’ can be more readily achieved in the winery by the use of nitrogen supplements, such as diammonium phosphate (DAP) and the choice of fermentation conditions. This point highlights the need to monitor nitrogen not only in the vineyard but also in the must immediately before fermentation, so that appropriate additions can be made when required. Overall, optimisation of vineyard and fermentation nitrogen can contribute to quality factors in wine and hence affect its value. However, a better understanding of the effect of nitrogen on grape secondary metabolites and different types of nitrogen sources on yeast flavour metabolism and wine sensory properties is still required.     

Effect of storage conditions on the volatile composition of wines obtained from must stabilized by PEF during ageing without SO2

The aim of this work was to study the effect of the storage conditions on the evolution of volatile composition of white wines aged in bottles without the addition of SO₂. Therefore Parellada must was stabilized by pulsed electric fields (PEF) and fermented without the employment of SO₂ and later on the wine was aged in bottles, without addition of this preservative, at low and controlled temperature and at room temperature. The obtained results showed that the concentration of some important compounds for wine aroma such as isoamyl acetate, and ethyl esters of fatty acids was higher in the wines stored at low and controlled temperature than in those aged at room temperature. However, the temperature favoured the formation of total alcohols during the aging of wines in the bottles. Consequently, from the point of view of the aromatic quality, the conservation of white wines obtained from fermentation of must processed by PEF and aged in bottles without the addition of SO₂ was improved under controlled storage conditions than at room temperature.

Industrial relevance

SO₂ is used as a preservative agent in wine due to its multi-action in the wine conservation. Although neither carcinogenicity nor mutagenic effects have been found in SO₂, this compound has an influence on human health. For that reason, several competent international organizations (WHO, FAO, OIV) have set down maximum limits for wines as well as promote a reduction of its concentration in wines. Therefore, potential industrial applications of this work include the possibility to produce and store wines without SO₂. This has been achieved by stabilizing the musts with PEF. The wines produced under these conditions can be conserved without this additive when used under controlled conditions of storage.     

Multivariate analysis for the differentiation of sparkling wines elaborated from autochthonous Spanish grape varieties: volatile compounds, amino acids and biogenic amines

The aim of this work was to focus on the study of the volatile composition of white and rosé base wines elaborated from different autochthonous grape varieties in Spain (Verdejo, Viura, Malvasía, Albarín, Godello, Prieto Picudo and Garnacha) and the evolution of the sparkling wines elaborated from them following the “champenoise” method. The amino acids and biogenic amines were also studied. Multivariate analyses (factorial and stepwise discriminant) were carried out to study the influence of these compounds all together. Base and sparkling wines from Albarín, Verdejo, Godello and Prieto Picudo were the richest in most of the volatile compounds analyzed, especially of ethyl esters and alcohol acetates, compounds that contribute to the fruity aroma of wines. During the aging on lees in the bottle of sparkling wines, an increase in branched ethyl esters, ethyl lactate and γ-butyrolactone, and a decrease in terpenes (mainly citronellol and geraniol) were observed. In general, the levels of biogenic amines in all the sparkling wines studied were very low, which does not represent a negative effect on their quality and safety.     

Simultaneous Quantification of Primary, Secondary Amino Acids, and Biogenic Amines in Musts and Wines Using OPA/3-MPA/FMOC-CI Fluorescent Derivatives

ABSTRACT: Free amino acids and biogenic amines occur in musts and wines and affect the course of fermentation in musts and the quality of wines. Each sample was analyzed in 138 min and each analysis involved quantification of 23 amino acids and 10 amines. The detection limit for proline was 27.3 mg/L. The mean detection limit for amino acids was 0.58 mg/L and for amines 0.28 mg/L. Precision was 0.5 to 19.2% for wine samples and accuracy by the standard addition method was 99.8% (recovery value). In all, 33 wines and 39 must samples were analyzed. In wines, ethanolamine, histamine, ethylamine, and putrescine amounted to 93% of the amine content.     

Influence of SO2 on the consumption of nitrogen compounds through alcoholic fermentation of must sterilized by pulsed electric fields

The aim of this work was to study the influence of SO₂ on the use of nitrogenous compounds by yeast during wine alcoholic fermentation. Thus Parellada must was sterilized by a pulsed electric field treatment and inoculated with Saccharomyces cerevisiae Na33 strain. The fermentations were carried out with SO₂ (20 mg/l) and without SO₂. Results showed that yeast better consumed the amino acids in the first half of fermentation in the presence of SO₂. The final concentration of amino acids in the obtained wine was greater when the must was fermented without SO₂ than when the latter compound was present. Therefore, the presence of SO₂ facilitated the consumption of amino acids and, hence, such wine should have more complex flavour and better microbiological stability than that obtained from fermentation without SO₂.     

Influence of addition of yeast autolysate on the formation of amines in wine

Amines, substances which can be toxic or have a negative effect on wine aroma, are formed during alcoholic and malolactic fermentations depending on various factors. In this work the influence of the addition of yeast autolysate to must on the concentration of amines in wine is studied. For this purpose Chardonnay must supplemented with the yeast autolysate was used; a second sample was also used without it (control sample). This autolysate provided amino acids and long‐chain fatty acids (C16 and C18). The enrichment of must by these nutrients did not produce an increase of amines during alcoholic fermentation despite the fact that consumption of amino nitrogen in general, and of some precursor amino acids in particular, was somewhat higher in the supplemented sample. After malolactic fermentation it was observed that the concentration of biogenic amines was higher in the wine from the supplemented must. Copyright © 2006 Society of Chemical Industry     

Malolactic fermentation of Tempranillo wine: contribution of the lactic acid bacteria inoculation to sensory quality and chemical composition

Malolactic fermentation (MLF) of Tempranillo Rioja wines (Spain) inoculated with two lactic acid bacteria (LAB) strains were studied and compared with spontaneous MLF. Inoculation with selected Oenococcus oeni lyophila shortened MLF duration up to 19 days and lead to wines with more fresh and fruity characters, especially when implantation was 100%. We demonstrated modifications in the concentration of volatile and nitrogenous compounds and a good correlation between analytical and sensory attributes was also noted. In addition, the low initial amino acid concentration and the consumption of these compounds by the inoculated yeast strain during alcoholic fermentation resulted in wines with very low biogenic amines levels (under 3.75 mg L^?1) after MLF and 3 month storage period in all cases. The results showed the significance of choose the most suitable starter to elaborate quality wines and suggest the control of amino acid content in must and wine to prevent the formation of biogenic amines.     

龙眼干白葡萄酒发酵过程中生物胺和氨基酸含量的变化

本研究利用高效液相色谱技术分析了龙眼干白葡萄酒酒精发酵和苹乳发酵过程中8种生物胺和22种氨基酸含量的变化。结果表明,在酒精发酵过程中8种生物胺含量均很低,而在苹乳发酵过程中迅速增加。发酵结束时干白葡萄酒中亚精胺和乙醇胺含量最高,其次为组胺和羟色胺。这4种生物胺的前体氨基酸在苹乳发酵阶段含量没有明显变化,而其它大部分氨基酸在酒精发酵过程中含量下降,在苹乳发酵过程中含量呈现不同程度升高。因此,我们认为：葡萄酒发酵过程中氨基酸含量变化与其对应生物胺的变化之间没有相关性,葡萄酒中生物胺的组成比例可能主要取决于乳酸菌中氨基酸脱羧酶的种类和活性。     

Biogenic amines in wines: Influence of oenological factors

Biogenic amines formation results from the decarboxylation of the corresponding amino acids by action of microorganisms. The presence of these compounds is considered by some authors a fundamental parameter for detriment of alcoholic beverages. The aim of this work was to assay the effect of some oenological factors (viticulture region, grape variety, anti-fungi treatment of grapes, fermentation activators, malolactic starters and storage on lees) from the point of view of their influence on the biogenic amines content of wines. According to our results, it was possible to show that the viticulture region affects the amounts of amines, since wines of some regions present higher contents of amines than wines from other regions. Grape varieties appear to influence the wine amines content. Commercial malolactic starters, after careful selection, should be added to the vinification process in order to decrease the formation of biogenic amines, since in our assays the wines that were inoculated with starters present lower amounts of biogenic amines. The wine storage on lees contributes for a biogenic amines increase.     

Influence of addition of ammonium and different amino acid concentrations on nitrogen metabolism in spontaneous must fermentation

The effect of the addition of different amino acid concentrations in must on yeast nitrogen metabolism during alcoholic fermentation was studied. To do this, fermentations of Mazuelo must, poor in nitrogen compounds, were carried out. Ammonium and different concentrations of amino acids (0, 45, 120, 250 and 450 mg/l) were added to the must. Addition of 45, 120 and 250 mg/l of proteic amino acids to the must increased the rate of fermentation. Proline was mainly consumed in fermentations with smaller amounts of amino nitrogen and, at the same time, this amino acid showed the highest residual concentration in the final wines. The consumption of other proteic amino acids was directly proportional to their concentration in the musts, with the exception of leucine and isoleucine that were synthesized. However, a difference in the percentages of the amino acids consumed by the yeasts was observed. The percentages of aspartic acid, alanine and arginine consumed were higher in the fermentations supplemented with amino acids than in the fermentation where only ammonium was added. The percentages of tyrosine and phenylalanine consumed gradually increased with increase of their initial concentration.     

Biogenic Amines in Wines

Biogenic amines are low molecular weight organic compounds that occur in fermented foods and are produced mainly through the microbial decarboxylation of amino acids. Decarboxylation of several amino acids can form some biogenic amines in wines such as histamine, tyramine, putrescine, and phenylethylamine. Biogenic amines can cause direct or indirect toxicity when their concentrations are high in food. The enological importance of the biogenic amines in wines is due to their possible toxicological risks and the possibility of the relationship between high amine content and unsanitary conditions during wine production. Various analytical techniques have been developed for the determination of biogenic amines in wines. In this article, the presence, the formation, the microorganisms and the methods of determination of biogenic amines in wine and other fermented foods are reviewed.     

Clarifying agents effect on the nitrogen composition in must and wine during fermentation

The effect of static sedimentation with and without clarifying agents (silica sol and gelatine) upon the nitrogenous fraction of musts and wines was studied. Four vinifications were carried out using a Vitis vinifera cv. Cayetana white grape must. Static sedimentation reduced less than other techniques the assimilable nitrogen (FAN), however the employment of fining agents promoted a net decrease. The changes in the amino acids during fermentation were similar in all the experiments carried out. In general, during the first days there was a fast decrease followed by a slight increment and then stabilisation. This decrease fitted, in most of the cases, with first order kinetics. For most of the amino acids, the percent consumption was higher in the must settled with clarifying agents. The clarifying agent’s addition did not have the same effect on the amino acid concentration in the final wines.     

Biogenic amines and the importance of starter cultures for malolactic fermentation

Biogenic amines are compounds produced mainly by microorganisms from the decarboxylation of some amino acids which may cause several health issues in susceptible people. In the winemaking process, one of the factors with the greatest impact on the final concentration of biogenic amines in wine is the malolactic fermentation, which mainly involves the action of lactic acid bacteria. This article gathers scientific literature on the importance of the use of inoculation in winemaking, especially for the potential formation of biogenic amines in spontaneous fermentations, with emphasis on malolactic fermentation. It establishes the set of attributes that influence the formation of biogenic amines in winemaking and aspects related to their toxicity, especially histamine and tyramine. With the growing interest of the market in products with less chemical appeal, the safety of wines, whether from non‐conventional or traditional winemaking, must increasingly be observed. Therefore, the importance of using cultivated starter cultures is discussed, along with the safety of organic wines and the presentation of articles that seek to reconcile the production of organic wine with the preservation of the ‘microbiological terroir’ in a safe way, using selected wild yeasts and lactic acid bacteria.     

Use of Sulfur Dioxide in Winemaking

Ten juices of common wine cultivars were studied to determine if pretreatment of the juice with air, blanketing the juice with nitrogen or pretreatment with sulfur dioxide (SO₂) was the most satisfactory method to make the better wine. Further treatment with SO₂ was made later at intervals. The pretreatment with aeration lowered the final color in the wine compared to nitrogen blanketing. SO₂ pretreatment gave the lowest colored juice for the white wines. All juices treated with SO₂ later had much lower color. Wines without SO₂ were generally less liked. Pretreatment of juice with SO₂ helped preserve good color and sensory attributes of the wines but later treatments with SO₂ were more essential in improving and maintaining quality. Oxidation was probably the most important control exercised by SO₂ additions.