Odors olfactometry

Fig. 3. (a) Flame ionization detector (fid) response to an extract of commercially processed Valencia orange juice, (b) Gas chromatography—olfactometry (geo) chromatogram of the same extract. The abscissa in both chromatograms is a normal paraffin retention index scale ranging between hexane and octadecane (Kovats index). Dilution value in the geo is the -fold that the extract had to be diluted until odor was no longer detectable at each index. 

The sensory technique used for assessing human perception of odors is called olfactometry. The basic technique is to present odorants at different concentrations to a panel of subjects and assess their response. The process favored by the U.S. National Academy of Sciences is dynamic olfactometry (16). This technique involves a sample dilution method in which a flow of clean, nonodorous air is mixed with the odorant under dynamic or constant... 

A laboratory where the measurement takes place must be free from odor and is typically air-conditioned with air filtration. The odor sample is placed in an olfactometer that basically is a device for dilution of the sample. Typically, the meter has two outlet ports diluted odorous air flows from one, and clean odor-free air flows from the other. In dynamic olfactometry, panel members assess the two ports of the olfactometer. The assessors indicate from which of the ports the diluted sample is flowing. The measurement starts with a dilution that is large enough to make the odor concentration beyond the panelists threshold. This concentration is normally increased by a factor of two in each successive presentation. Only when the correct port is chosen and the assessor is certain that the choice is correct and not just a guess, is the response considered a true value. 

The last set of requirements in olfactometry is concerned with the differences between panel members. People vary widely in their sensitivity. A factor of a 100 between the thresholds of two subjects for the same substance is not uncommon. For a number of substances, specific anosmia s or specific hyposmia s are found. In such cases a person has no sensitivity at all or a very high threshold for the given substance, but normal sensitivity to other substances (1). This is an illustration of the fact that sensitivity to odours is specific rather than general. This is also demonstrated by Punter (2, 3) who determined the thresholds of 69 odorous substances for the same group of subjects and calculated the correlations between these thresholds (see figure 2). 

Although knowledge on the correlation of odorous compounds concentration and odour impression is still limited, it is used in all types of olfactometry. Indeed diluting this concentration by adding pure air is a general practice. Also many investigations were performed where chemicals are added to air and used in psychophysical experiments. Many speakers in this workshop will present data in this field. Here only chemical analysis will be dealt with. 

New, powerful techniques in chemistry, odor formulations, bioassays, and olfactometry have supplied us with deeper as well as fresh insights into olfactoiy effects on our behavior. Medicine, psychology, environmental design, occupational safety, air-quality control, marketing, and advertising now consider and contribute to human chemical ecology. 

Lee, S.-J. Noble, A. C. Characterization of Odor-Active Compounds in Californian Chardonnay Wines using GC-Olfactometry and GC-Mass Spectrometry. J. Agric. Food Chem. 2003, 51, 8036-8044. 

Mistry, B.S., Reineccius, T., Olson, L.K. (1997) Gas chromatography olfactometry for the determination of key odorants in foods. In Marsili R (ed) Techniques for Analysing Food Aroma. Dekker, New York, pp 265-292. 

Eyres, G., Dufour, J.P> Hallifax, G., Sotheeswaran, S., Marriott, P.J. (2005) Identification of character-impact odorants in Coriander and wild coriander leaves using gas chromatography-olfactometry (GCO) and comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GCxGC-TOFMS). J. Sep. Sci. 28 1061-1074. 

The extract volume at which the odorant was most (AECA) or least (AEDA) perceived by gas chromatography-olfactometry... 

Milo, C. and Grosch, W. 1995. Detection of odor defects in boiled cod and trout by gas chromatography-olfactometry of headspace samples. J. Agric. Food Chem. 43 459-462. 

It should be kept in mind that most analytical instruments, such as gas chromatographs and mass spectrometers, do not discriminate between volatile compounds that do or do not possess odor activity. Some form of sensory analysis must be conducted in order to select which volatile compounds contribute to the flavor of the foods. Gas chromatography-olfactometry (GC/O) is an important tool to accomplish that task. 

In the early history of gas chromatogra-phy/olfactometry (GC/O vn/tgu), the goal of GC/O analysis was to determine when an odor elutes from a GC in order to identify it. The analysis yielded a list of times and, with appropriate standards, retention indices. When combined with other chemical analysis methods, such as mass spectrometry (MS), a name for a particular odorant could be proposed. Comparing both the chemical and sensory properties of the odorant with those of authentic standards allowed researchers to identify the odorant with considerable certainty. The number of odorants that are detected, however, is determined by a number of factors, including the design of the olfactometer, the fraction of the extract injected, and, as we now suspect, the genetics of the sniffer. 

A first approach to distinguish between the odor-active compounds and the many odorless volatiles present in such aroma extracts is the application of gas chromatography/olfactometry (GCO, formerly called "sniffing-technique" [13-17]). 

Gas chromatography/olfactometry (GCO) methods have been developed as screening procedures to detect potent odorants in food extracts. The FD-factors or CHARM values determined in food extracts are not consequently an exact measure for the contribution of a single odorant to the overall food flavor for the following reasons. During GCO the complete amount of every odorant present in the extract is volatilized. However, the amount of an odorant present in the headspace above the food depends on its volatility from the food matrix. Furthermore, by AEDA or CHARM analysis the odorants are ranked according to their odor thresholds in air, whereas in a food the relative contribution of an odorant is strongly affected by its odor threshold in the food matrix. The importance of odor thresholds in aroma research has been recently emphazised by Teranishi et al. [58],... 

The method of choice for the analysis of odorants and subsequent identification is gas chromatography-olfactometry (GC—O), a method which combines the separation capability of volatile compounds by GC with the selective and sensitive odor detector human nose (Fuller, Steltenkamp and Tisserand, 1964). A scheme of the set-up can be seen in Figure 8.6. 

Odor analysis (see Chapter 8) performed using GC coupled with olfactometry has also shown that many food items and household materials are odorant sources (Mayer and Breuer, 2006). Thus, mono-unsaturated aldehydes particularly E-2-nonenal are found in fat, wax, oil finish and lubricants branched aldehydes such as 3-methyl butanal are found in varnish, bread and malt while leather, rice and popcorn are sources of substituted pyrrolines especially 2-acetyl-l-pyrroline. Studies like this are important not only from the point of view of identifying sources of indoor odorants but also from the point view of providing vital information that can help consumers to select products. 

In addition to GC-MS, recent studies have focused on the identification and quantitative analysis of impact odorants in botrytized wines using gas chromatography-olfactometry (GC-O) analysis. Sarrazin et al. (2007a) investigated numerous botrytized and nonbotry-tized Sautemes wines. They could identify several key odorants that were responsible for the sensory differences between the wines, notably 3-mercaptohexan-l-ol, various furanons, ethyl-hexanoate, methional, phenylethanol, phenylacetaldehyde, sotolon, p-damascenone, and 2-methyl-3-furanthiol. 

Odor-active components in cheese flavor, many of which are derived from milk lipids, can be detected using GC-olfactometry (GC-O). GC-0 is defined as a collection of techniques that combine olfactometry, or the use of the human nose, as a detector to assess odor activity in a defined air stream post-separation using a GC (Friedich and Acree, 1988). The data generated by GC-0 are evaluated primarily by aroma extract dilution analysis or Charm analysis. Both involve evaluating the odor activity of individual compounds by sniffing the GC outlet of a series of dilutions of the original aroma extract and therefore both methods are based on the odor detection threshold of compounds. The key odourants in dairy products and in various types of cheese have been reviewed by Friedich and Acree (1988) and Curioni and Bosset (2002). 

Curioni, P.M.G., Bosset, J.O. 2002. Key odorants in various cheese types as determined by gas chromatography-olfactometry. Int. Dairy J. 12, 959-984. 

B. S. Mistry T. Reineccius L. K. Olson, Gas Chromatography-Olfactometry for the Determination of Key Odorants in Foods. In Techniques for Analyzing Food Aroma R. Marsili, Ed. Marcel Dekker New York, 1997 pp 265-292. 

The standard measurement procedure for odor determination (VDI, 1994) is called olfactometry. It uses the human olfactory sense (Gostelow et al, 2001) for the determination of odor qualities. The human nose is an extremely sensitive odor detector and is used in subjective and objective sensory measurements. The latter expresses the strength of odor in terms of the number of dilutions of odor-free air required to reduce the sample odor to threshold concentration (Gostelow et al, 2001). The threshold concentration is reached when the human nose can just smell the odorous substance. Because every nose has different sensitivity, the standard test procedure involves four people at the same time. Prior to olfactory... 

Source of odor Volume flow rate V/m ft- Olfactometry odor/ou m % Odor reduction... 

Aznar, M., Lopez, R., Cacho, J.F., Ferreira, V. (2001). Identification and quantification of impact odorants of aged red wines from Rioja. GC-olfactometry, quantitative GC-MS, and odor evaluation of HPLC fractions. J. Agric. Eood Chem., 49, 2924-2929. 

The second point is related to the simultaneous presence of odorants at g/L levels and of others that can be active at levels as low as ng/L. This means that although it makes sense to use a general screening procedure for detecting by olfactometry the potentially most relevant aroma molecules, it will not be possible to use a single isolation or preconcentration scheme to identify and further quantify the different aroma molecules. Rather, it will be necessary to have an array of chemical isolation and quantification procedures if a comprehensive aroma analysis is our objective. 

In conclusion, the study of the wine aroma chemicals and the understanding of the role they play in the different wine aroma nuances have to be structured into a numbers of steps strongly constrained by the previous considerations. Such steps will be the subject of thischapter. The first step is about the screening of aroma molecules, which will be carried out by using gas chromatography-olfactometry. The second will be the isolation and identification of odorants. The third is the quantitative determination, for which only a very brief outline will be given, and the fourth is about the sensory tools used to assess the sensory role played by the different odorants. 

Since all aroma molecules are more or less volatile, the technique that a priori is best suited to screen the odor active molecules from the rest of molecules is Gas Chromatography-Olfactometry (GC-O). This technique makes use of the human nose as detector for the compounds eluting out of the chromatographic column, typically a fused silica capillary column (Acree et al. 1984). There are several different approaches for GC-O differing in the way in which the olfactometric signals are... 

Etievant, P.X., Callement, G., Langlois, D., Issanchou, S., and Coquibus, N. (1999). Odor intensity evaluation in gas chromatography - olfactometry by finger span method. J. Agric. Food Chem., 47, 1673-1680. 

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