terclim by ICS banner
IVES 9 IVES Conference Series 9 INVESTIGATION INTO MOUSY OFF-FLAVOR IN WINE USING GAS CHROMATOGRAPHY-MASS SPECTROMETRY WITH STIR BAR SORPTIVE EXTRACTION

INVESTIGATION INTO MOUSY OFF-FLAVOR IN WINE USING GAS CHROMATOGRAPHY-MASS SPECTROMETRY WITH STIR BAR SORPTIVE EXTRACTION

Abstract

Mousy off-flavor is one of the defects of microbial origin in wine. It is described as a particularly unpleasant defect reminiscent of rodent urine (a “dirty mouse cage”), and grilled foods such as popcorn, rice, crackers, and bread crust. Prior to the 2010s, mousiness was very uncommon but it has been becoming more frequent in recent years. It is often associated with an increase in pH as well as certain oenological practices, which tend to significantly decrease the use of sulfur dioxide.

Three major compounds, 2-acetyl-1-pyrroline (APY), 2-acetyltetrahydropyridine (ATHP) and 2-ethyltetrahydropyridine (ETHP), have been identified as responsible for mousiness in wines. A particularity of these compounds is that they are prone to tautomerism and can coexist in several forms. Moreover, the nitrogen atom in the heterocyclic ring can be protonated under specific conditions, i.e. with pH lower than pKa, inducing a positive charge and, as a consequence, an increase of polarity and a loss of volatility of the molecule.

To date quantification data reported in the literature are limited due to analytical issues related to the nature of these compounds. To fill the gap and later understand the parameters influencing mousiness, the objective of this study was to develop a simple and effective method to simultaneously determine trace levels of these three mousy N-heterocycles in wines. Therefore, a stir bar sorptive extraction (SBSE) followed by GC-MS analysis was developed (1).

Firstly, both previously reported tautomers of ATHP (2), 2-acetyl-1,4,5,6-tetrahydropyridine and 2-acetyl-3,4,5,6-tetrahydropyridine were identified, unlike to APY and ETHP. The extraction conditions were then optimized paying particular attention to the pH of the sample. The performance of the developed method was evaluated on white, rosé and red wines and the limits of detection and quantification of the method are lower than previously published concentrations in spoiled wine.

The method was then applied to provide quantitative data by analyzing 6 control wines and 68 wines produced without added sulfites. ETHP was detected in almost all wines produced with limited use of SO₂. ATHP was detected in almost all wines suspected of mousiness whereas APY was only detected in few cases. This method will provide a support for further studies aimed at understanding the phenomena that influence the occurrence of mousy off-flavor and the oenological parameters that modulate its expression.

DOI:

Publication date: February 9, 2024

Issue: OENO Macrowine 2023

Type: Poster

Authors

Céline Franc¹, Daiki Kiyomichi², Pierre Moulis1,3, Laurent Riquier¹, Patricia Ballestra¹, Stéphanie Marchand¹, Sophie Tempère1 and Gilles de Revel¹

1. Univ. Bordeaux, INRAE, Bordeaux INP, Bordeaux Sciences Agro, UMR 1366, OENO, ISVV, F-33140 Villenave d’Ornon, France
2. Institute for Future Beverages, Kirin Holdings Company, Limited, 4-9-1 Jonan, Fujisawa, Kanagawa 251-0057, Japan
3. Department of Microbiology and Biochemistry, Hochschule Geisenheim University, Geisenheim, Germany

Contact the author*

Keywords

mousiness, N-heterocycles, quantification, SBSE-GC-MS

Tags

IVES Conference Series | oeno macrowine 2023 | oeno-macrowine

Citation

Related articles…

CHANGES IN METABOLIC FLUXES UNDER LOW PH GROWTH CONDITIONS: CAN THE SLOWDOWN OF CITRATE CONSUMPTION IMPROVE OENOCOCCUS OENI ACID-TOLERANCE?

Oenococcus oeni is the main Lactic Acid Bacteria responsible for malolactic fermentation, converting malic acid into lactic acid and carbon dioxide in wines. Following the alcoholic fermentation, this second fermentation ensures a deacidification and remains essential for the release of aromatic notes and the improvement of microbial stability in many wines. Nevertheless, wine is a harsh environment for microbial growth, especially because of its low pH (between 2.9 and 3.6 depending on the type of wine) and nutrient deficiency. In order to maintain homeostasis and ensure viability, O. oeni possesses different cellular mechanisms including organic acid metabolisms which represent also the major pathway to synthetize energy in wine.

WINE LEES AS A SOURCE OF NITROGEN FOR OENOCOCCUS OENI TO IMPROVE MALOLACTIC FERMENTATION PERFORMANCE

Malolactic fermentation (MLF) is a desired process in red and acidic white wines, after alcoholic fermentation (AF), carried out by the lactic acid bacterium (LAB) Oenococcus oeni. The advantages are an increase of pH, microbiological stabilization and organoleptic improvement of the final wine. However, the presence of stress factors such as ethanol, low pH, high total SO2, lack of nutrients and presence of inhibitors, could affect the successful completion of MLF [1]. Changes in amino acid composition and deficiencies in peptides after AF, showed that MLF can be delayed, signaling its importance for bacterial growth and L-malic acid degradation during MLF [2].

EVALUATING WINEMAKING APPLICATIONS OF ULTRAFILTRATION TECHNOLOGY

Ultrafiltration is a process that fractionates mixtures using semipermeable membranes, primarily on the basis of molecular weight. Depending on the nominal molecular weight cut-off (MWCO) specifications of the membrane, smaller molecules pass through the membrane into the ‘permeate’, while larger molecules are retained and concentrated in the ‘retentate’. This study investigated applications of ultrafiltration technology for enhanced wine quality and profitability. The key objective was to establish to what extent ultrafiltration could be used to manage phenolic compounds (associated with astringency or bitterness) and proteins (associated with haze formation) in white wine.

PROTEOMIC STUDY OF THE USE OF MANNOPROTEINS BY OENOCOCCUS OENI TO IMPROVE MALOLACTIC FERMENTATION

Malolactic fermentation (MLF) is a desired process to decrease acidity in wine. This fermentation, carried out mostly by Oenococcus oeni, is sometimes challenging due to the wine stress factors affecting this lactic acid bacterium. Wine is a harsh environment for microbial survival due to the presence of ethanol and the low pH, and with limited nutrients that compromise O. oeni development. This may result in slow or stuck fermentations. After the alcoholic fermentation the nutrients that remain in the medium, mainly released by yeast, can be used in a beneficial way by O. oeni during MLF.

STATISTICAL COMPARISON OF GROWTH PARAMETERS OF NINE BIOPROTECTION STRAINS IMPLEMENTED ON ARTIFICIALLY CONTAMINATED SYNTHETIC MUST

In recent years, consumer demand for products without chemical additives increased, becoming a priority for the wine sector. SO₂ is widely used for its multiple properties including antiseptics, antioxidants and antioxidasics and the strategy of bioprotection in winemaking represents now an alternative to this chemical additive. In oenology, results have highlighted the interest of bioprotection to limit the development of microorganisms like Hanseniaspora uvarum and thus reduce the doses of sulphite. Indeed, this species is considered because of its acetic acid and methyl butyl acetate production, the latter can cover the varietal character of wines.