Journal of the Science of Food and Agriculture

J Sci Food Agric 85:23912396 (2005) DOI: 10.1002/jsfa.2240

The electronic nose as a tool for the classication of fruit and grape wines from different Ontario wineries
Robin C McKellar,1 HP Vasantha Rupasinghe,2 Xuewen Lu3 and Kelley P Knight1
1 Food

Research Program, Agriculture and Agri-Food Canada, Guelph, Ontario N1G 5C9, Canada of Environmental Sciences, Nova Scotia Agricultural College, Truro, Nova Scotia B2N 5E3, Canada 3 Department of Mathematics and Statistics, University of Calgary, Calgary, Alberta T2N 1N4, Canada
2 Department

Abstract: Electronic nose technology is useful for classifying or ngerprinting foods and beverages based on odour proles. With a view to providing useful information on quality attributes, the Fox 3000 electronic nose (EN) was tested for the ability to characterize Ontario-produced fruit wines. Eight fruit wines (blueberry, cherry, raspberry, blackcurrant, elderberry, cranberry, apple and peach) and four grape wines (red, Chardonnay, Riesling and ice wine) were each obtained from a minimum of ve Ontario wineries. Replicates of each wine sample were dried onto membrane lters to remove ethanol, and analyzed by the EN. It was possible to separate completely each wine variety (eg blueberry) based on differences between wineries; however, when all wine data were pooled, classication by variety was poor (58.7% correctly classied). Analysis of different wine varieties from a single winery revealed some misclassication. Wines could be separated into four distinct groups based on position on the discriminant function analysis map (79.9% correct). Fruit and grape wines were well separated from each other (75.9% correct), as were red and white wines (92.2% correct). The results show that the EN can discriminate fruit and grape wines into natural and useful groupings and may become an important tool for standardization of wine quality. 2005 Society of Chemical Industry

Keywords: fruit wine; electronic nose; discriminant function analysis (DFA); standardization

INTRODUCTION The electronic nose (EN) is an analytical instrument designed to mimic the human nose.1 4 It has been applied extensively in the chemical and cosmetic industries but it has only recently been used to address problems in the food and beverage industry.3 There are a number of different technologies available but, generally, an EN consists of a semi-selective sensor array which reacts with volatile chemicals in the sample headspace. In our laboratory we have used a commercial EN (Fox 3000, AlphaMOS, Toulouse, France) which consists of 12 metal oxide sensors, each of which has a particular specicity for a class of compounds, such as hydrocarbons, aldehydes, amines and aromatics, depending upon the type and amount of semiconducting lm which has been applied. As ushed gases from a sample are passed over the sensors, the reaction between oxygen, the lm and volatile molecules results in a change in conductivity of the sensor through oxidation (increases conductance) or reduction (decreases conductance) reactions to form a

pattern.2 The patterns are then processed and analyzed using advanced pattern-recognition techniques such as principal components analysis (PCA), or discriminant function analysis (DFA). A database of known samples can be created, and the EN can then be used to identify unknown samples by comparison with the database with a high degree of accuracy.5 We have used the Fox 3000 EN in a number of applications, including the development of orange juice avour,6 the detection of spoilage in packaged minimally processed lettuce,7 and the optimization of beer ageing.8 One of the most challenging and potentially useful applications of EN technology is classication and quality assurance of wines. Sensory and chemical properties of wine, especially colour, aroma and taste, are in part related to the total concentration and prole of avonoids (mainly anthocyanins and avonols), but may also be inuenced by products produced by the yeast during fermentation. Characterization of wine aroma is a complex and difcult problem, and the development of more rapid methods is desirable.

Correspondence to: Robin C McKellar, Food Research Program, Agriculture and Agri-Food Canada, 93 Stone Road West, Guelph, Ontario N1G 5C9, Canada E-mail: mckellarr@agr.gc.ca Contribution number S167 from the Food Research Program (Received 16 April 2004; revised version received 4 January 2005; accepted 17 February 2005) Published online 13 July 2005

2005 Society of Chemical Industry. J Sci Food Agric 00225142/2005/$30.00

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Over the past decade, there have been a number of attempts to apply EN technology to wines, with varying degrees of success. Because of the wide variety of technologies and approaches, it is difcult to summarize the progress effectively or to make valid comparisons between the various studies. The EN devices employed have been largely non-commercial units, based generally on metal oxide sensor technology.9 16 Other technologies such as conducting polymers17 19 and quartz microbalances20,21 have seen limited use. Two commercial units have been used: the AromaScan (based on conducting polymers)19 and the Fox 4000 (based on metal oxide sensors).12 Wines have been analyzed from various regions in either Spain17,18,22 or Italy.15,16 Wines have been classied by variety or denomination,11,15 18,21,23 class (red/white/ros ),15 18 vintage11,16 or vineyard.9,11,20 e Other applications include degree of ageing,22 winemust fermentation19,24 and degree of oak wine barrel toasting.12 The high concentration of alcohol in wines is the factor which most strongly limits the application of EN technology. Alcohol swamps the sensors, and makes it difcult, if not impossible, to differentiate wines.15 19,25 Separation of alcohol from the volatiles of interest prior to EN analysis is desirable and methods to achieve this include solid-phase micro-extraction (SPME),18,22 pervaporation19,24 and dynamic headspace sampling using a resin.17 These methods are often complex and require extensive sample manipulation, thus the simple approach of drying samples on a membrane lter was adopted in our laboratory.8,26 While some volatiles might be lost, it was still possible to discriminate beer aged for different periods of time using this method.8 Non-traditional fruit wines are gaining in popularity in Canada but have yet to be characterized for successful marketability. Fruit wine producers are anxious to market their products based on the quality attributes with comparison to traditional wines. The avour and aroma of fruit wines are distinct from the source of fruit used for the fermentation. There are no recorded applications of EN technology to the classication of fruit wines, thus the objectives of the present study were (1) to assess the ability of the FOX 3000 electronic nose to distinguish between fruit wines on the basis of variety and winery, and (2) to characterize the Ontario-produced fruit wines with comparison to traditional grape wines (white, ice wine and red wine).

same species (Vitis vinifera) as the Chardonnay and Riesling. The ice wines were all Vidal, a hybrid cultivar. For each variety of wine, single bottles were taken from ve or six Ontario wineries during 2002. It was not possible to nd wineries which produced all 12 wine types so, in order to have a minimum of ve wineries for each wine type, a total of 33 wineries participated in the study. The vintage years ranged from 1998 to 2002 and alcohol content ranged from 9% to 14.5% (v/v). Operation of the electronic nose Five replicate samples (250 L) of each fruit or grape wine were spotted onto separate 25 mm, 0.45 m nylon membrane lters (Fisher Scientic) and allowed to air dry for 2 h. The membranes were placed into 10 mL autosampler vials (Fisher Scientic) and capped with silicon/Teon magnetic autosampler vial caps (London Scientic). Each vial was loaded into an autosampler (HS50, CTC Analytics, Switzerland) and heated to 75 C for 8 min. Headspace (2500 L) was removed from each vial and injected into the EN with a 30 min delay between samples to allow the sensors to return to the baseline. Data analysis The raw sensor data were analyzed by DFA after being divided up into different databases consisting of: individual wine varieties from up to six producers; all fruit and grape wine data; fruit and grape wine separately; and wines divided into four groups based on DFA analysis of all wines by variety. Raw output from each of the 12 sensors was analyzed by DFA using StatTools, a Microsoft Excel add-in (Palisade Decision Tools, Neweld, NY). DFA is typically used to determine which continuous variables (here the sensor outputs) discriminate between two or more naturally occurring groups. DFA provides orthogonal (independent) linear discriminant functions such that the rst one provides the most overall discrimination between groups, the second provides the second most, and so on. In practice, two discriminant functions (here designated LD1 and LD2) are generally sufcient to describe most of the variation. The intention is to minimize the within-group differences, and maximize the betweengroup differences. The common practice is to use discriminant functions to attempt to predict group membership of samples which were not used in the development of the original model. In the absence of such data, cross-validation can be performed. Each sample is removed sequentially from the model, and the model reformulated without that sample. The classication of the removed sample is then tested against the new model. Cross-validations are presented in the present study.
J Sci Food Agric 85:23912396 (2005)

EXPERIMENTAL Source of wine samples Eight varieties of fruit wine (blueberry, cherry, raspberry, blackcurrant, elderberry, cranberry, apple and peach) and four varieties of grape wine (red, Chardonnay, Riesling and ice wine) were examined in this study. The red wines were either Cabernet-Merlot, Cabernet Sauvignon or Cabernet Franc, all the
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RESULTS Initially, each of the individual wine varieties was analyzed by DFA to see if they could be discriminated on the basis of winery. The analysis for blueberry wine is shown in Fig 1 as an example. Figure 1(a) shows a cluster plot of each of the ve replicates (from a single bottle) for each winery. Three of the clusters were well separated, while the other three mapped close together (bottom left corner of Fig 1). All the wineries were sufciently well separated so that cross-validation placed 100% of the samples with the correct winery. For all 12 wine types, this analysis correctly classied 100% of the samples (data not shown). It became apparent that using cluster plots as shown in Fig 1(a) would present difculties when analyzing large numbers of samples. Consequently, the centroid of each group was calculated, with the standard deviation for both linear discriminants as error bars. This gives some indication of the data scatter; 95% of the data points should fall within two standard deviations of the centroid point. Figure 1(b) shows the data in Fig 1(a) replotted with error bars, further demonstrating the distinction between wineries. The output of the DFA for all wine varieties discriminated by variety is given in Table 1. The wine varieties in the rst column are the original classications, with the other columns giving the new classication based on the DFA analysis. For example, reading horizontally, the rst row shows that, of the 30 apple samples, 16 were classied by DFA as apple, two as cherry, two as cranberry, three as elderberry and ve as ice wine. Considering the number of wine varieties being classied, a reasonably good job was done by the DFA, with an overall percentage correct of 58.7%. Peach had the highest number of samples (19 of 25) correctly classied. There was a tendency for blueberry and cherry to be misclassied as each other. Similarly, the Chardonnay/Riesling pair were also misclassied. Interestingly, other wine varieties were also misclassied as Chardonnay or Riesling, in particular peach.
Table 1. DFA and cross-validation of all wines by type

20

(a) 1 2

10

3 4 5

LD2

10 20 (b)

10

10 10 5 0 5 LD1
Figure 1. Discriminant function analysis (DFA) of blueberry wine from six different wineries showing (a) ve replicate analyses of each wine, and (b) standard deviations of LD1 and LD2.

10

15

20

The somewhat limited ability of the EN to discriminate between different wine varieties may have been due, at least in part, to the inter-winery differences. Since most wineries produced at most three or four different wine types, it was difcult to assess the extent of confounding which could be attributed to the wineries. As an example, one winery was selected which produced blueberry, cherry, raspberry and apple wines. DFA analysis gave 90% discrimination, with only one each of ve cherry and blueberry replicates being misclassied as the other.

Classication Blackmatrix Apple currant Blueberry Chardonnay Cherry Cranberry Elderberry Ice wine Peach Raspberry Red Riesling Apple Blackcurrant Blueberry Chardonnay Cherry Cranberry Elderberry Ice wine Peach Raspberry Red Riesling 16 2 1 0 0 1 3 0 0 0 0 1 2 18 0 0 0 0 0 0 0 2 0 0 0 1 14 0 7 1 1 1 0 0 0 0 0 3 0 16 2 2 0 0 3 1 4 7 2 0 11 0 19 0 1 0 0 6 0 0 2 0 0 1 0 21 5 1 0 0 5 5 3 0 0 0 0 0 15 0 0 0 0 0 5 1 2 1 1 0 0 18 0 1 0 1 0 0 0 0 0 0 0 0 19 0 0 4 0 2 0 0 0 0 0 4 0 18 0 0 0 1 2 3 0 0 0 0 0 1 18 2 0 2 0 9 1 5 0 0 3 1 3 10

Initial assignments are shown in the rst column and classication by DFA is given in the rst row. For example, of 30 apple samples, DFA classied them into 16 apple, 2 blackcurrant, 2 cherry, 2 cranberry, 3 elderberry and 5 ice wine.

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Raspberry, blackcurrant, cranberry and apple wines produced by another winery were 100% separated by DFA analysis. These ndings are in agreement with the classications of all wine varieties found in Table 1. The results in Table 1 are also supported by the plots of group centroids for all wine varieties (Fig 2; error bars omitted for clarity). The various wine varieties were visually separated into four arbitrary groups based on position in the gure. Those wines with centroids: LD1 > 1 (blueberry, cherry, raspberry and ice wine) were designated group 1; LD2 > 1 (apple, blackcurrant, elderberry) were designated group 2; 1 < LD1 < 1 (Chardonnay, red, Riesling, cranberry) were designated group 3; LD1 < 2 (peach) were designated group 4. In order to further examine the association among these groups, the three groups with more than one member (groups 1, 2 and 3) were plotted separately, with standard deviations for LD1 and LD2 given as error bars (Fig 3). These plots show that there was considerable overlap in standard deviation within each of the three groups. More specically, the blueberry/cherry and Chardonnay/Riesling wine pairs had centroids which were very close, and, with the expected scatter in the data, were difcult to separate. The validity of the above four groups was tested by attempting to classify all of the wine varieties into these groups by DFA. Table 2 shows that overall 79.9% of the samples were classied correctly, suggesting that these four groups might form a reasonable classication system for wines based on volatile chemicals. Further analysis of the wine groupings in Fig 2 was done on separate databases for each of the three groups with two or more members. Table 3 gives the classication for groups 1, 2 and 3. The least discrimination was achieved with group 1 (70.2% correct), with improvements for group 2 (77.6% correct) and group 3 (75.8% correct). As
3.5 2.5 1.5 LD2 0.5 0.5 1.5 2.5 3 5 4 3 2 1 0 LD1 1 2 1 3 4 4 2 Red Riesling Apple Blackcurrant Blueberry Chardonnay Cherry Cranberry Elderberry Peach Raspberry Ice wine

(a)

Blueberry Cherry Raspberry Ice wine

1 2 0.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 0.5 0.5 0.0 0.5 1.0 1.5 2.0 2.5 2.0 1.5 1.0 0.5 LD1
Figure 3. DFA of wines by variety from (a) group 1, (b) group 2 and (c) group 3. Bars indicate standard deviations.

0.5

1.0

1.5

2.0

2.5

3.0 Apple Blackcurrant Elderberry

(b)

LD2

3 (c)

2 Red Riesling Chardonnay Cranberry

0.0

0.5

1.0

Figure 2. DFA of all wines by variety separated into four groups. Table 2. DFA and cross-validation of all wines by group

Classication matrix Group 1 Group 2 Group 3 Group 4

See note for Table 1.

Group 1 100 14 11 0

Group 2 3 57 5 0

Group 3 8 13 97 4

Group 4 3 1 7 21

noted above, it was difcult to separate blueberry and cherry wines and Chardonnay and Riesling, even though a reduced variability would be expected since the data for the three groups was analyzed separately. Three of the four grape wines were members of Group 3 (Fig 2), suggesting that there might be signicant differences between classes (fruit or grape) of wine. In this case, a good (75.9% correct) classication was achieved. It was more common for fruit wines to be misclassied as grape (60 of 230) than for grape wines to be misclassied as fruit (5 of 114). Attempts were made to further classify wines based on subsets of the original dataset. For example, wine varieties were divided into two distinct datasets (fruit or grape) and analyzed separately. Discrimination of fruit wines by variety was not very successful, with percentage correct values of <70%. On the other hand, when grape wine varieties were classied into either red or white wine, classication was successful with a percentage correct of 92.1%. Of the red wines, 26 of 30 were correctly classied, while 79 of 84 of the white wines were correct.

DISCUSSION In this study we report the rst successful discrimination of fruit and grape wines using an
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Table 3. DFA and cross-validation of three groups by type

Group 1 Blueberry 16 8 2 1 Raspberry 0 4 0 26 22 4 5

Group 2 Blackcurrant Elderberry

Group 3 Cranberry

Classication matrix Group 1 Blueberry Cherry Ice wine Raspberry Apple Blackcurrant Elderberry Chardonnay Cranberry Red Riesling

Cherry 11 17 1 2

Ice wine 3 1 21 1

Apple

Chardonnay

Red

Riesling

Group 2

4 24 0

4 2 20 20 2 3 5 1 25 0 0 0 3 21 0 9 0 6 25

Group 3

See note for Table 1.

EN. Specically, each of the 12 different varieties of wines (eg blueberry, Riesling) when analyzed separately could be clearly separated by winery. In other words, the EN could distinguish substantial differences in volatile patterns which were characteristic of each winery or wine-making procedure. This suggests a potential role for the EN in the standardization of fruit wines and provides the means to establish quality attributes which will result in increased economic benet to producers. The EN was less able to separate the 12 varieties of wines from each other, in part owing to the differences between wineries. However, analysis of wine varieties within single wineries revealed that there were still sufcient similarities between different fruit wines to result in some misclassication. The observed similarities between wines prompted a division of the various wine varieties into four groups based on their position on the DFA map in Fig 2. Interestingly, all of the grape wines except ice wine were in group 3, as was cranberry wine. The separation of ice wine from the other grape wines may have been due in part to the fact that all of the ice wine samples were of the Vidal variety, a different species from the other grape wines. Peach wine was in a group by itself, but mapped near group 3 (Fig 2), suggesting that peach and cranberry wines may have some characteristics in common with grape wines. There are limited published studies on the successful use of EN technology to classify wines. Many of the studies have focussed on the development of the sensor technology and optimization strategies, rather than practical applications. An EN based on metal oxide sensor technology has been used to discriminate Italian red wines by variety (or denomination), vintage and vineyard.9,11 Separation of different white23 and red17 wine was also achieved; however, in these studies, an additional pretreatment step was required to maximize separation. One of the common limitations is that alcohol interferes strongly with most EN systems, thus much effort has been expended in either removing the alcohol or selectively extracting
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the specic wine volatiles. We have adopted a simple approach, where small aliquots of wine were dried on the surface of lters. This technique was used in earlier studies where spoiled and unspoiled wine were correctly identied26 and the degree of ageing of beer was determined.8 The present study adds support to the idea that good discrimination of wine can be achieved by the EN with minimal sample preparation. The aggregation of three of the four grape wines into a single group suggests important differences between grape and fruit wines. DFA was able to discriminate all 12 wine varieties into two classes, grape or fruit wine. Interestingly, it was more common for fruit wines to be misclassied as grape during cross-validation than misclassication of grape wines as fruit, again suggesting similarities between fruit and grape wines. Wines in group 3 appeared to be particularly closely related, with Chardonnay and Riesling wines often being misclassied as each other. Analysis of the four grape wine varieties together also indicated that red wine was easily separated from the other wine types.

CONCLUSION The present study has clearly shown that the EN has high potential for the characterization and discrimination of fruit and grape wines. Strong inter-winery differences exist which may be used to characterize a particular winery. Further experiments are required to establish standards to allow application of this approach in a commercial setting. The EN can also distinguish differences in the volatile proles of closely related wine types, and has established important relationships between wines from different fruits. The ability of processors to exploit close relationships between some fruit wines and white grape wines as determined by the EN may have some marketing potential. Thus, the EN may have the ability to elucidate the relationship between wines, and ensure quality and uniformity based on aroma components, but further studies are warranted.
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15 Penza M and Cassano G, Chemometric characterization of Italian wines by thin-lm multisensors array and articial neural networks. Food Chem 86:283296 (2004). 16 Penza M and Cassano G, Recognition of adulteration of Italian wines by thin-lm multisensor array and articial neural networks. Anal Chim Acta 509:159177 (2004). 17 Guadarrama A, Fernandez JA, Iniguez M, Souto J and De Saja JA, Array of conducting polymer sensors for the characterization of wines. Anal Chim Acta 411:193200 (2000). 18 Guadarrama A, Fernandez JA, Iniguez M, Souto J and De Saja JA, Discrimination of wine aroma using an array of conducting polymer sensors in conjunction with solidphase micro-extraction (SPME) technique. Sens Actuators B 77:401408 (2001). 19 Pinheiro C, Rodrigues CM, Schafer T and Crespo JG, Monitoring the aroma production during wine-must fermentation with an electronic nose. Biotechnol Bioeng 77:632640 (2002). 20 Di Natale C, Paolesse R, Macagnano A, Mantini A, DAmico A, Ubigli M, Legin A, Lvova L, Rudnitskaya A and Vlasov Y, Application of a combined articial olfaction and taste system to the quantication of relevant compounds in red wine. Sens Actuators B 69:342347 (2000). 21 Di Natale C, Paolesse R, Burgio M, Martinelli E, Pennazza G and DAmico A, Application of metalloporphyrins-based gas and liquid sensor arrays to the analysis of red wine. Anal Chim Acta 513:4956 (2004). 22 Rodriguez-Mendez ML, Arrieta AA, Parra V, Bernal A, Vegas A, Villanueva S, Gutierrez-Osuna R and De Saja JA, Fusion of three sensory modalities for the multimodal characterization of red wines. IEEE Sens J 4:348354 (2004). 23 Heberle I, Liebminger A, Weimar U and Gopel W, Optimised sensor arrays with chromatographic preseparation: Characterization of alcoholic beverages. Sens Actuators B 68:5357 (2000). 24 Sch fer T, Bengtson G, Pingel H, B a oddeker KW and Crespo JPSG, Recovery of aroma compounds from a wine-must fermentation by organophilic pervaporation. Biotechnol Bioeng 62:412421 (1999). 25 Privat E, Roussel S, Grenier P and Bellon-Maurel V, Techniques for ethanol removal before discrimination of alcoholic drinks using electronic noses (A review). Sci Aliment 18:459470 (1998). 26 McKellar RC, Cliff M, Smith M and Knight K, Monitoring the quality of wine using the electronic nose. Annual Meeting of the Canadian Institute of Food Science, Kelowna, BC, Canada (1999).

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