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- Acetic acid and ethyl acetate in Madeira wines: evolution with ageing and assessment of the odour rejection thresholdPublication . Miranda, Andreia; Pereira, Vanda; Pontes, Marisela; Albuquerque, Francisco; Marques, José C.Madeira wine is an internationally recognized fortified wine, characterized by a complex flavour and high longevity. Little is known about the impact of volatile acidity in its features along ageing. Thus, this work studied the evolution of acetic acid and ethyl acetate (volatile acidity main contributors) during the ageing processes used in Madeira wine production, canteiro and estufagem. Wines aged in canteiro for 6 up to 45 years were also evaluated. The odour rejection thresholds (ORT) were assessed in sweet (Malvasia) and dry (Sercial) wines with 5 and 10 years old, by a regular consumer panel and a non-regular consumer panel. The results showed that the formation trend of acetic acid and ethyl acetate is similar in both ageing processes and is favoured by the ageing, mostly in sweet wines. The ORT of ethyl acetate was in average 328 mg/L for both panels, while for acetic acid ranged between 1.96 and 5.72 g/L, depending on the evaluation panel and also on the age and sweetness degree of the Madeira wine. The ORT values expressed in volatile acidity ranged between 1.4 and 3.5 g/L, which are higher than the legal limits. The study pointed out that the olfactory perception of acetic acid and ethyl acetate in Madeira wines depends on their age and sweetness degree.
- Analytical methodologies for the determination of biogenic amines in wines: an overview of the recent trendsPublication . Miranda, Andreia; Leça, João M.; Pereira, Vanda; Marques, José CarlosBiogenic amines are naturally present in grapes or can occur during the vinification and aging processes, essentially due to the microorganism’s activity. When present in wines in high amount, biogenic amines may cause not only organoleptic defects but also adverse effects in sensitive human individuals, namely due to the toxicity of histamine, tyramine and putrescine. Even though there are no legal limits for the concentration of biogenic amines in wines, some European countries only recommend maximum limits for histamine. In this sense, biogenic amines in wines have been widely studied. The determination of amines in wines is commonly achieved by liquid chromatography, using derivatization reagents in order to promote its separation and detection. In alternative, other promising methodologies have been developed using capillary electrophoresis or biosensors, revealing lower costs and faster results, without needing a derivatization step. Nowadays, it is still a challenge to develop faster and inexpensive techniques or methodologies to apply in the wine industry. Thus, this review will be focused on the studies published in the last decade that involves the determination of biogenic amines in wines, highlighting the novelty, improvement and optimization of the analytical methods. The sample preparation procedures (such as derivatization reagents), the analytical methodologies and the new trends being followed by the wine industry are also described and discussed.
- Impact of indigenous non-saccharomyces yeasts isolated from Madeira Island vineyards on the formation of ethyl carbamate in the aging of fortified winesPublication . Leça, João Micael; Pereira, Vanda; Miranda, Andreia; Vilchez, José Luis; Malfeito-Ferreira, Manuel; Marques, José CarlosThe impact of selected non-Saccharomyces yeasts on the occurrence of ethyl carbamate (EC) was evaluated. Hanseniaspora uvarum, Starmerella bacillaris, Pichia terricola, Pichia fermentans and Pichia kluyveri isolated from Madeira Island vineyards were inoculated in Tinta Negra musts. Urea, citrulline (Cit) and arginine (Arg) were quantified when the density of musts attained the levels to obtain sweet (1052 ± 5 g/L) and dry (1022 ± 4 g/L) Madeira wines. The urea concentration varied between 1.3 and 5.3 mg/L, Cit from 10.6 to 15.1 mg/L and Arg between 687 and 959 mg/L. P. terricola and S. bacillaris generated lower levels of urea (<2.5 mg/L), Cit (<11.0 mg/L) and Arg (<845.6 mg/L). The five resulting fortified wines, individually fermented by the selected non-Saccharomyces yeast, were exposed to laboratory-accelerated aging at 70 °C for 1 month. From the studied yeasts, P. terricola and S. bacillaris revealed a lower potential to form EC (<100 µg/L); therefore, both yeasts can be a useful tool for its mitigation in wines.
- New insights into ethyl carbamate occurrence in fortified winesPublication . Leça, João M.; Pereira, Vanda; Miranda, Andreia; Vilchez, José Luis; Marques, José C.The occurrence of ethyl carbamate (EC) in fortified wines was studied testing different model wines under accelerated ageing (45 °C for 4 months and 70 °C for 1 month), to investigate the possibility of arginine (Arg) being a direct precursor of EC in fortified wines, comparing it with known major precursors, urea and citrulline (Cit). Wine main sugars were appraised as possible catalysts, as previously pointed out. Model wines showed that: Arg can induce the EC formation under accelerated ageing, even without being metabolised by microorganisms; although much less reactive, Arg can play a significant role since high residual levels can occur in young fortified wines; glucose (Glc) and fructose (Fru) suppressed the EC formation from urea and Cit pathways, in about 11–26%. Moreover, Madeira wine samples were investigated in order to appraise the eventual contribution of the alcoholic fortification. The results revealed that this step can promote a decrease of the amount of these EC precursors up to 46%. Despite preliminary, additional information about the EC formation in fortified wines was obrained namely for designing new mitigation strategies, which can pass through the reduction of residual Arg.
- Olfactory perception threshold assessment of volatile acidity in Madeira winesPublication . Miranda, Andreia Fátima Santos; Marques, José Carlos Antunes; Pereira, Vanda Nulita GomesMadeira wine is a fortified wine with impact in the Madeira Island’s economy. Similarly to other wines, its acidity should be well controlled in order to ensure Madeira wine quality, mostly the volatile acidity. Due to Madeira wine complex flavour, it is crucial to get a better knowledge about the volatile acidity impact in its features, namely determine the perception limit of acetic acid and ethyl acetate, as both are the main contributors for volatile acidity. Firstly, the olfactory perception threshold of volatile acidity was assessed by a trained and an untrained panel, using 5 and 10 years-old Sercial and Malvasia wines. Moreover, the current work also presents the evolution of organic acids, acetic acid and ethyl acetate during 540 days of ageing of Madeira wines (Malvasia, Bual, Verdelho and Sercial), comparing the same wines aged by both traditional ageing processes: canteiro and estufagem. Other wine samples, aged in wood casks (canteiro) for at least 5 years, were also evaluated. HS-SPME followed by GC-MS analysis was used to determine ethyl acetate concentration and IEC-HPLC-DAD was used for the organic acids determination, including acetic acid. The results indicated that acetic acid and ethyl acetate olfactory perception threshold depends essentially on wine’s age. Concerning acetic acid, the untrained panel was in average 5.45 g/L (5 years-old) and 6.22 g/L (10 years-old). Training the expert panel to recognize acetic acid odour, the values decreased for 1.44 g/L (5 years-old) and 1.87 g/L (10 years-old), but still remained higher than the established volatile acidity legal limits. Ethyl acetate threshold was similar for both panels (in average 327.97 mg/L). Both compounds tend to increase exponentially with age, being more evident in sweet wines. Organic acids in young Madeira wines depend mostly on the nature of grape varieties, but this difference is minimized with wine ageing.
- Impact of Non-Saccharomyces Yeast Fermentation in Madeira Wine Chemical CompositionPublication . Miranda, Andreia; Pereira, Vanda; Jardim, Humberto; Malfeito-Ferreira, Manuel; Marques, José CarlosMadeira wine is produced via spontaneous alcoholic fermentation arrested by ethanol addition. The increasing demand of the wine market has led to the need to standardize the winemak ing process. This study focuses on identifying the microbiota of indigenous yeasts present during Madeira wine fermentation and then evaluates the impact of selected indigenous non-Saccharomyces as pure starter culture (Hanseniaspora uvarum, Starmerella bacillaris, Pichia terricola, Pichia fermentans, and Pichia kluyveri) in the chemical and phenolic characterization of Madeira wine production. Re sults showed that the polyphenol content of the wines was influenced by yeast species, with higher levels found in wines produced by Pichia spp. (ranging from 356.85 to 367.68 mg GAE/L in total polyphenols and 50.52 to 51.50 mg/L in total individual polyphenols through HPLC methods). Antioxidant potential was higher in wines produced with Hanseniaspora uvarum (133.60 mg Trolox/L) and Starmerella bacillaris (137.61 mg Trolox/L). Additionally, Starmerella bacillaris stands out due to its sugar consumption during fermentation (the totality of fructose and 43% of glucose) and 15.80 g/L of total organic acids compared to 9.23 g/L (on average) for the other yeasts. This knowledge can be advantageous to standardizing the winemaking process and increasing the bioactive compounds, resulting in the production of high-quality wines.
- Modelling the ageing process: a novel strategy to analyze the wine evolution towards the expected featuresPublication . Pereira, Ana C.; Carvalho, Maria J.; Miranda, Andreia; Leça, João M.; Pereira, Vanda; Albuquerque, Francisco; Marques, José C.; Reis, Marco S.In this work we present a new strategy to monitor the wine evolution during the ageing process. More specifi cally, we validate a procedure for analyzing how wine evolves during the ageing process in relation to the desired and expected quality features and we apply the proposed methodology to the case of a Portuguese fortified wine, the Madeira wine, where we compare the wine evolution under two different ageing processes. The approach developed consists on modeling samples labeled as aged reference wines (5 year old Madeira wines), produced from four different grape varieties, and then analyze how and in which extent young wines (up to 3 years old) come closer to the reference data set. The analysis is based on a comprehensive set of chemical data, including: polyphenolic composition, organic acids, reducing sugars, color and oenological parameters, commonly used as routine quality control information. The study considers several feature extraction methods, such as: Principal Components of Analysis (PCA), Independent Component of Analysis (ICA) and Partial Least Squares (PLS). The classification methodologies tested were: Linear Discriminant Analysis (LDA), nearest neigh bor (k-NN) and Soft Independent Modelling by Class Analogy (SIMCA). The different options of preprocessing/ feature extraction/classification were evaluated and compared using a Monte Carlo approach. From our analysis, the best combination of feature extraction/classification methodologies was PLS/LDA, which presented a classification performance of approximately 90% for three out of the four classes modeled, and of about 78% for the remaining one. Regarding the wines monitored during the first 3 years, our analysis revealed that they indeed mature in relation to the five year old reference wines. Furthermore, for some wines, it is possible to detect differences between the two ageing processes analyzed. This study is of particular importance for this type of wines, where the ageing process plays a central role for attaining the expected quality levels, implying significant risks and costs for local and industrial producers. Notwithstanding the specific case study presented, the strategy outlined can be extrapolated to other products with similar characteristics in terms of their monitoring and process control
- Indigenous yeasts in Madeira wines: impact on the physicochemical and sensorial characterizationPublication . Miranda, Andreia Fátima Santos; Marques, José Carlos AntunesO Vinho Madeira é produzido pela fermentação espontânea através de leveduras indígenas presentes em uvas e adegas. A crescente exigência dos consumidores leva à necessidade de uniformizar o processo de vinificação. O presente trabalho visa avaliar a composição química de vinhos Madeira produzidos em diferentes adegas e com uvas colhidas de diferentes localizações (sul e norte). Depois, isolar e identificar as leveduras indígenas de mostos da casta Tinta Negra de diferentes localizações e adegas, e avaliar o impacto do uso de 5 leveduras indígenas não-Saccharomyces selecionadas (Hanseniaspora uvarum, Starmerella bacillaris, Pichia terricola, Pichia fermentans e Pichia kluyveri) na produção de vinho Madeira, como culturas puras iniciadoras, na composição química, perfil volátil e características sensoriais. Os resultados mostraram variabilibidade na composição química nas diferentes adegas, e baixa reprodutibilidade entre lotes da mesma adega. Uvas da região sul apresentaram níveis mais elevados de açúcares (196.32 g/L), polifenóis totais (270.18 mg GAE/L) e potencial antioxidante (128.48 mg Trolox/L). Os estudos da microflora de leveduas indígenas envolvidas na produção de vinho Madeira permitiram identificar 11 espécies nas adegas (Hanseniaspora uvarum, Starmerella bacillaris, Pichia kluyveri, Pichia fermentans, Saccharomyces cerevisiae, Torulaspora delbrueckii, Candida apicola, Cystobasidium minutum, Pichia terricola, Cystobasidium slooffiae, e Wicheramomyces anolalus) e 6 espécies nas uvas (Hanseniaspora uvarum, Starmerella bacillaris, Pichia kluyveri, Pichia fermentans, Saccharomyces cerevisiae, and Hanseniaspora opuntiae). Os vinhos produzidos por Pichia spp. demonstraram níveis mais elevados de polifenóis totais (367.68 mg GAE/L), enquanto os vinhos produzidos por Starmerella bacillaris demonstraram maior acidez (9.23 g/L) e menor conteúdo de voláteis (11.26%). O vinho produzido por Pichia kluyveri destacou-se na concentração total de volatéis (24.50%), dos quais 77.35% são estéres, obtendo 37% de preferência dos consumidores, com notas intensas de caramelo e especiarias. Portanto, Pichia kluyveri demonstrou potencial para ser utilizada como cultura iniciadora de fermentação na produção de vinho Madeira.