E. T. Contis et al.

(Editors)
Food Flavors: Formation, Analysis and Packaging Influences
© 1998 Elsevier Science B.V. All rights reserved 117

Comparisons of volatile compounds released during consumption of

Cheddar cheeses by different consumers

CM. Delahunty, P.J. O'Riordan, E.M. Sheehan and P.A. Morrissey

Department of Nutrition, University College, Cork, Ireland

Abstract
Methods exist for measuring volatile compounds released in the mouth during food
consumption, however little work has compared the volatile compounds released during
consumption by different consumers or related individual differences to consumers' chewing
patterns and saliva production rates. In this work, eight consumers were chosen and each
consumed six Cheddar cheeses during Buccal Headspace Analysis (BHA). Released volatile
compounds were measured for each cheese and for each consumer. Electromyography was
used to record each consumers chewing style, and their saliva production rate was also
measured. It was found that although there were differences in consumers' chewing styles
and saliva production rates, the volatile profiles obtained by BHA, for each individual, were
similar for each cheese when compared with the other cheeses examined.

1. INTRODUCTION

It is the volatile compounds of a food, released in sufficient concentration during

consumption, which stimulate the olfactory epithelium and induce perceived odor. Recent
flavor research has emphasised the importance of volatile release from a food matrix and
shown how volatile release is related to consumer flavor perception. This work is often
driven by a food industry which must reduce costs or meet the demands of diet conscious, but
discerning, consumers who wish to reduce fat and salt intake. Substitution for these
ingredients is necessary to restore removed flavor and regulate flavor release.
However, understanding of flavor release is made difficult by the complexity of the
interactions between foods and consumers. Each volatile compound has different physio-
chemical properties and its release is influenced by interactions with other food matrix
variables such as moisture, fat, protein, carbohydrate, and other soluble (salt, sugars) and
non-soluble materials. In addition, food breakdown and mixing with saliva during
consumption, respiratory air flow over and around the food and temperature and pH changes
occurring in the consumers mouth will influence volatile release and subsequent flavor
perception. It is also known that individual consumers expression of flavor differs as a result
of physiological, psychological and social differences [1,2]. Therefore an underlying question
which remains unanswered is; what part of flavor differences between foods result from
volatile release dynamics from the food matrix, and what part result from differences between
consumers? Conclusions reached in response to this question have been mixed.
118

There are model systems which measure volatile compounds released while mimicking
conditions in the mouth [3-5]. Other methods measure volatile release directly during
consumption using mass spectrometry of breath [6,7] and indirectly by trapping volatiles on
adsorbents, such as Tenax, before analysis [8-10]. Soeting and Heidema [6] showed thirty-
fold differences in the relative quantities of 2-pentanone which was measured directly from
the breath of different consumers. Van Ruth et al [10] also found subject specific volatile
profiles were released during consumption of vegetables. Taylor et al [11] trapped volatiles
released from mint sweets during consumption and also found differences between subjects
in terms of the quantities of volatiles released. However, they concluded that there were
similarities between the relative concentrations of volatiles released for each subject.
Delahunty et al [12], who analyzed Buccal Headspace Analysis [BHA;13] data using
Principal Components Analysis [PCA;14] to examine the volatile profiles released during
consumption of cheeses by three different consumers, found product specific volatile release
was most important. However, three consumers were too few to draw any firm conclusions.
Workers studying food texture have developed methods such as electromyography [15]
which measure muscle activity during mastication of a food matrix. From these
measurements they have shown mastication patterns and can calculate the amount of work
done by a consumer during consumption. These methods, which show considerable
differences between consumers' mastication characteristics, have recently been related to
differences between consumers' temporal perception of flavor intensity measured by time-
intensity sensory analysis [16]. Other physiological parameters, such as the influence of
saliva [4] and air flow through the mouth [17] have also been investigated.
The present study was carried out to investigate discrepancies in the literature relating to
the differences between consumers' interactions with foods and the relationships found
between physiological measures during chewing and individuals' differences in flavor
perception. In order to achieve this, similar varieties of a complex food were chosen and a
multivariate technique, PCA, was used to examine the volatile profiles released.

2. EXPERIMENTAL

2.1. Samples and consumers

Six Cheddar cheeses, in 5kg blocks, of equal age (6-8 months) were obtained from 4
different producers. Eight consumers, 3 female and 5 male, aged between 22 and 28 were
used for all studies.

2.2. Buccal Headspace Analysis

Buccal Headspace Analysis of each cheese was carried out for each consumer in
triplicate. For this method a 50 g cheese sample was consumed in 10 x 5g pieces in a normal
way, allowing 30 s for the consumption of each piece. During the entire consumption time (5
min) volatile compounds released were displaced through the nose by vacuum and trapped on
a Tenax-TA trap. The order of sample analysis was balanced for consumers, cheeses and day
of consumption [18]. A blank buccal headspace sample was taken each day for each
consumer. Traps were thermally desorbed using a Teckmar Purge and Trap 3000 concentrator
(Teckmar, Cincinnati, OH, USA). Desorbed volatiles were identified and quantified using gas
chromatography-mass spectrometry (GC-MS) with a Varian Saturn GC-3400CX
 119

incorporating a Varian Saturn 3 GC/MS detector (Varian chromatography systems, Mitchell

drive, Walnut Creek, CA, USA).The column was a DB-5ms, 30m x 0.257mm fused silica
capillary column, with a film thickness of 0.25 |Lim (J & W scientific, Folsom, CA, USA)

2.3 Mastication behaviour

The activity of the consumers left and right masseter muscles during chewing was
recorded by Electromyography [15]. The electromyograph record was measured for 1 cheese
over a period of 5 min (10 x 30 s for each 5g piece ), in triplicate, for each of the eight
consumers. Each individuals electromyogram was integrated using a poly VIEW data
acquisition and analysis system (Grass instrument division, Astro-Med Inc., East Greenwich
Avenue, West Warwick, UK)

2.4 Saliva production

Consumers unstimulated saliva production was measured by allowing their saliva to drip
into a beaker for a 5 min period. The consumers swallowed immediately before collection
and forcefully spat out at the end [19]. The stimulated saliva production was measured by
dividing the volume of saliva produced by each consumer in response to 50g of cheese (10 x
5g) by the chewing time required by the consumer for that cheese [19]. Each measurement
was repeated four times.

2.5 Data Analysis

Buccal headspace data was analyzed by PC A, using the Unscrambler v 6.0 (CAMO AS,
N-7041 Trondheim, Norway), of the log transformed peak areas of volatile compounds.
Electromyography data was analyzed by Analysis of Variance (ANOVA) using SPSS v 6.1
(SPSS Inc. Chicago, IL 60611, USA) of the totals for chew number, chew time, chew rate
and chew work. Saliva production data was analyzed by ANOVA of the unstimulated and
stimulated saliva flow rates. Differences between cheeses and between subjects were
investigated using ANOVA. Relationships between data sets were investigated by linear and
Partial Least Squares regression [PLS;20], using the Unscrambler v 6.0.

3. RESULTS AND DISCUSSION

In the present study the quantities and balance of volatile compounds released during
consumption of a food, by different consumers, was compared. For this purpose Cheddar
cheese was chosen as this represents a complex protein matrix containing fat and moisture.
To minimize product related compositional differences, and therefore to maximize the
influence of consumer related differences to volatile release from one food type, cheeses of
equal age were chosen. Eighteen volatile compounds were selected from chromatograms of
BHA of all cheeses and the amounts of each present were quantified. Both Figures 1 and 2
depict two PCA's. The first (in italics) was calculated using individual consumers' headspace
data (triplicates averaged) and the second using the average of the 8 consumers.
120

Figure 1. PC A scores on PC's 1 and 3 for 6 cheeses assessed by BHA using 8 subjects (A-H)
(see text for explanation).The pooled SD for the analysis is represented by an ellipse on
cheese 1.

4 T

3B
4C 5C
6E
13H "^ fr ^^ Ic IG
^^ ,2iP2f 4H 2F
c 4G -im 4A IB 2C
o 3G 5F
a. -4

OH
2 6D
5

6G

-4 -^

Principal component 1 {?>9Vo)(24%)

Principal Components (PC) 1 and 3, which accounted for 39% and 15% of the explained
variance, respectively, of the PC A of compound peak areas, showed significant differences {p
= 0.017 and p = 0.021) between cheeses (Table 1 and Figure 1). Differences between
consumers (p = 0.050), which accounted for \3% of the explained experimental variance,
were found on PC3 (Table 1 and Figure 2).

Table 1
ANOVA between cheeses (1 - 6 ) and between consumers ( subjects A • H)on
Principal Components 1 - 4 of the PC A
Principal component Cheese Subject
PCI 0.017 0.873
PC2 0.657 0.072
PC3 0.021 0.050
PC4 0.055 0.351

The volatile compounds which distinguished the cheeses from one another on these
components are shown in the PC loadings plots (Figures 3 and 4).The differences found
between cheeses on PCI, which was the most important as it contained the highest proportion
of the experimental variance (39%), were caused mostly by the quantities of compounds
released during consumption by each consumer rather than by their balance.
 121

Figure 3. PC A loadings for 18 volatile compounds on PC's 1 and 3 for 6 cheeses assessed by
BHA using 8 subjects (A-H).

0.5 T
2-heptanone

cyclohexanel
cyclohexane2

cpdl3
cpdl4
o
OH
cpd3
-0.5 cpd5 0.5
toluene cpd2
dmds heptane

cpdl7
(Eodecane
ethyl butyrate
cpdl2

cpdl6
-0.5 -^

Principal component 1 (39%)

However, differences between cheeses found on PC's 2 and 3 were caused mostly by
differences in the balance of the compounds released. This can be determined from the
relative positions of the volatile compounds in the loadings plots (Figure 4).
In a previous study physiological differences between consumers have been related to
differences in flavor perception [16]. In this study very significant differences were found
between consumers mastication characteristics and also between their saliva production rates
(p = 0.000 for all parameters apart from chew rate (p = 0.021) and chew work (p = 0.045))
(Table 2 and Figure 5). Using linear regression chew number and chew work were found to
relate to saliva production rate during cheese consumption for 5 of the 8 subjects (r = 0.98
and r = 0.84, respectively). However by using PLS and linear regression, no significant
relationships were found between the measured physiological characteristics and total volatile
release. Sensory evaluation of the cheeses is not reported in this study and therefore no
conclusions can be made with regard to consumers' expressions of flavor perception. Further
work is also required to investigate the dynamics of volatile release during time of
consumption.

4. CONCLUSION

Some differences were found between the quantities of volatile compounds released during
cheese consumption by different consumers. Very significant differences were found between
consumers mastication characteristics and between saliva production rates during cheese
consumption. Despite these differences, the distribution of experimental variance explained
122

Table 2
Mastication behaviour and saliva production rates of 8 consumers ( subjects A-H)

Subject Electromyography^ Saliva flow rate''

TCN TCT CR CW Unstim. Stimulated

A 224.33 141.26 1.59 2435.86 0.77 5.30

B 138.67 169.07 1.08 1011.45 0.43 4.06
C 348.00 212.27 1.67 3823.63 0.59 1.69
D 324.33 198.53 1.64 4497.6 1.06 7.31
E 228.67 144.00 1.68 2066.55 0.53 4.52
F 215.67 212.07 1.02 1194.73 0.70 4.89
G 184.33 145.71 1.27 866.71 0.76 3.25
H 282.33 180.99 1.55 3010.14 0.35 3.48
pooled SD 22.84 28.35 0.12 1403.50 0.12 0.65
P 0.000 0.000 0.021 0.045 0.000 0.000
^ TCN = total chew number; TCT = total chew time (sec); CR = chew rate (chew / sec); CW = chew work
b Unstim. = unstimulated saliva production rate (mL / min); Stimulated saliva production rate (mL / min)

Figure 5. Mastication behaviour of 3 consumers during consumption of one 5 g piece of

cheese. CN = chew number; CT = chew time (sec); CR = chew rate (chew / sec); CW = chew
work.

Pat (A)
CN = 23
CT= 14.66
CR=1.57
C W = 133.03

Chew Time (sec)

 123

Figure 3. PCA scores on PC's 2 and 3 for 6 cheeses assessed by BHA using 8 subjects (A-H).
The pooled SD for the analysis is represented by an ellipse on cheese 6.

4 T

4C
2A C ID
IG ifP
6C 2B 2F 3D
o r^
OH 4A 2C
5E 5G
6F
5D
ex 6Bry
3F6D

,^

-4 -^

Principal component 2 (\3%)(]2%)

Figure 4. PCA loadings of 18 volatile compounds on PC's 2 and 3 for 6 cheeses assessed by
BHA using 8 subjects (A-H).

0.5 T
2-heptanone

cyclohexanel
cycldhexane2
pctane
cdd7 cpdl2
cpdl4
o
a. |cpd3
-0.5 cpf 0.5
cpd2 toluene
dmds heptane

cpdl7
dodecane
ethylbutyrate
cpdl2

cpdl6
-0.5 -^

Principal component 2 (13%)

124

by the PCA showed that Cheddar cheese of equal age could be identified by their product
specific volatile release. Therefore, the volatile profile for a particular cheese at the end of
consumption was found to be similar in all consumers.

5. ACKNOWLEDGEMENT

This work was part funded by the Department of Agriculture, Food and Forestry, Ireland,
under the Food Industry Sub-Programme of EU Structural Funds.

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