Chem. Senses 25: 131-140,
2000
© Oxford University Press 2000
Synergism among Ternary Mixtures of Fourteen Sweeteners
Susan S. Schiffman,
Elizabeth A. Sattely-Miller,
Brevick G. Graham,
Barbara J. Booth1 and
Kernon M. Gibes1
Department Of Psychiatry, Duke University Medical Center, Durham, NC 27710 and
1 NutraSweet-Kelco Co., Mt Prospect, IL 60056, USA
Correspondence to be sent to: Susan S. Schiffman, Department of Psychiatry, Box 3259, Duke University Medical Center, Durham, NC 27710. e-mail: sss{at}acpub.duke.edu
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Abstract
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The purpose of the present study was to determine the degree
of synergism of sweet taste among ternary mixtures of 14 sweeteners.
A trained panel evaluated ternary mixtures of 14 sweeteners
varying in chemical structure and type. The ternary mixtures
that were tested were limited to those in which the compounds
comprising the mixture were synergistic in binary combinations,
according to an earlier study. All sweeteners in the ternary
mixtures were isointense with 2% sucrose, according to a previously
developed formulae. Each self-mixture was also tested (e.g.
2% sucrose + 2% sucrose + 2% sucrose). The triad with the highest
mean sweetness intensity rating was alitameneohesperidin
dihydrochalconerebaudioside-A (10.8). This represents
an increase of 99.4% when compared with the average of the self-mixtures.
While this is greater than the maximum of 74% increase found
for binary mixtures, more dyadic combinations of sweeteners
tested previously exhibited synergism than ternary combinations
tested here. However, most ternary mixtures were synergistic
(significantly greater than the average of the three self-mixtures)
to some degree.
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Introduction
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Synergistic taste interactions occur for some sweetener combinations
such that the total sweetness intensity of a mixture is greater
than the theoretical sum of the intensities of the individual
components (Bartoshuk, 1975

; Bartoshuk and Cleveland, 1977

;
Frank
et al., 1989

; Ayya and Lawless, 1992

; Schiffman
et al.,
1995

; Birch, 1996

, 1999

; Hutteau
et al., 1998

; Lawless, 1998

).
In a study of binary combinations of 14 sweeteners, two factors
found to influence whether a mixture would exhibit synergy were:
(i) the presence of a high potency sweetener in the mixture
and (ii) the concentrations of the component sweeteners in the
mixture. For some sweeteners, mixtures of two components at
concentrations that were isosweet with 3% sucrose were more
likely to exhibit synergy than mixtures of two components that
were isosweet with 5% sucrose or 7% sucrose. For other sweeteners,
the converse was true. Overall, the presence of a high potency
sweetener in the binary mixture produced more synergistic effects
than the presence of sugars (Schiffman
et al., 1995

). In another
study of binary mixtures of sweeteners, synergy was reported
to be influenced by water structure and the nature of hydration
of the sweetener molecules (Hutteau
et al., 1998

).
The purpose of the present study was to evaluate ternary mixtures of the same sweeteners found to produce synergy in binary combinations by Schiffman et al. (Schiffman et al., 1995
). The goal was to determine if the blend of three sweeteners in a mixture provides a greater or lesser synergistic effect than the blend of two sweeteners.
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Materials and methods
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Subjects
A trained panel of 18 subjects, ten females and eight males, participated in the study. The maximum number of subjects participating in any given taste session was 18 and the minimum number was nine. All subjects were from either the Duke University or Durham, NC communities. Their mean age was 46 ± 15 years. All subjects were paid for their participation.
Stimuli
Fourteen sweeteners were tested in the study: three sugars (fructose, glucose, sucrose); two polyhydric alcohols (mannitol, sorbitol); two terpenoid glycosides (rebaudioside-A, stevioside); two dipeptide derivatives (alitame, aspartame); one sulfamate (sodium cyclamate); one protein (thaumatin); two N-sulfonylamides (acesulfame-K, sodium saccharin); and one dihydrochalcone (neohesperidin dihydrochalcone).
Procedure
Ternary mixtures of 14 sweeteners were tested with a trained panel. Sweeteners in every mixture were at concentrations determined to be isointense with 2% sucrose, according to formulae determined by DuBois et al. (DuBois et al., 1991
) (see Table 1). Not all ternary combinations of sweeteners were tested. The ternary mixtures that were tested were limited to those for which the three sweeteners in the mixture were synergistic in binary combinations according to an earlier study by Schiffman et al. (Schiffman et al., 1995
). Each self-mixture of all 14 sweeteners was also tested (e.g. 2% sucrose + 2% sucrose + 2% sucrose).
Prior to evaluating the mixtures, each trained panelist tasted
six sweet taste references according to the method described
by DuBois
et al. (DuBois
et al., 1991

): 2 sweet (2% sucrose),
5 sweet (5% sucrose), 7.5 sweet (7.5% sucrose), 10 sweet (10%
sucrose), 12 sweet (12% sucrose) and 15 sweet (16% sucrose).
These sucrose standards have been used previously in a variety
of studies on sweeteners (Schiffman
et al., 1994

, 1995

; Portmann
and Kilcast, 1996

; Hutteau
et al., 1998

). Panelists also tasted
bitter references labeled 2.2 bitter (0.02% caffeine) and 4
bitter (0.03% caffeine), and sour references labeled 2.1 sour
(0.01% citric acid) and 7.4 sour (0.08% citric acid). Bitter
and sour references were based upon previous evaluations by
the present panelists, as well as other trained panelists.
At any given taste session, subjects provided sweetness intensity ratings, as well as other flavor profile notes, for five ternary mixtures. The five mixtures tested on any given day were randomized in accordance with a random number table. Panelists received 15 ml of each mixture in 30 ml plastic medicine cups which were labeled with a random three-digit number. The order of presentation of the five samples chosen at any given taste session was randomized across panelists. After tasting all references and rinsing their mouths with deionized water, panelists would swirl the unknown samples around in their mouths before expectorating. Panelists would then perform a full flavor profile of the sample, including all tastes, aromatics and feeling factors. In doing a flavor profile, panelists would make a mark on a 15 cm line scale that was anchored at 0, 5, 10 and 15 cm, and would then measure the length of the mark with a ruler. These marks reflected the intensities perceived for each flavor note by the individual panelist. Subjects also indicated the time of onset of maximum sweetness intensity by circling either early, middle or late. Between evaluations of the five sweeteners on any given test day, panelists would rinse their mouths with deionized water and eat unsalted top crackers in order to eliminate lingering tastes in their mouths. Subjects refrained from smoking, eating or drinking anything but water for 30 min prior to each panel session.
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Results
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Table 2 gives the least squares mean of each triad, as well
as the associated 95% two-sided confidence interval. An analysis
of variance (ANOVA) was conducted comparing the (least squares)
mean of any given triad with the average of its three constituent
sweetener self-mixtures. This method of determining synergism
is discussed in an earlier paper (Schiffman
et al., 1995

). Also
included in
Table 2 is the numerical difference in the two aforementioned
means, which is under the column headed Synergy estimate
and the corresponding
P-value of the
t-test against 0
(Synergy
P-value). For the self-mixtures, the
synergy estimate and
P-value columns are not applicable. Fifty-six
of the 79 combinations containing three different sweeteners
showed synergism. Of the 23 combinations that were not synergistic,
21 contained at least one bulk sweetener (e.g. fructose, glucose,
mannitol, sorbitol, sucrose).
Figures 1
14 show the least
squares mean sweetness intensity ratings of each ternary combination
containing acesulfame-K, alitame, aspartame, fructose, glucose,
mannitol, Na cyclamate, Na saccharin, neohesperidin dihydrochalcone,
rebaudioside-A, sorbitol, stevioside, sucrose and thaumatin,
respectively. A dashed line is given to indicate a nominal response
(i.e. an additive response).

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Figures 114 Figure 1 The least squares mean sweetness intensity ratings of every ternary combination tested containing acesulfame-K, alitame, aspartame, fructose, glucose, mannitol, Na cyclamate, Na saccharin, neohesperidin dihydrochalcone, rebaudioside-A, sorbitol, stevioside, sucrose and thaumatin, respectively. A dashed line is given to indicate a nominal response (i.e. an additive response).
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Figures 114 Figure 14 The least squares mean sweetness intensity ratings of every ternary combination tested containing acesulfame-K, alitame, aspartame, fructose, glucose, mannitol, Na cyclamate, Na saccharin, neohesperidin dihydrochalcone, rebaudioside-A, sorbitol, stevioside, sucrose and thaumatin, respectively. A dashed line is given to indicate a nominal response (i.e. an additive response).
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Table 3 gives a comparison of the responses of the ternary mixtures
in this study with the responses of the binary mixtures in a
previous paper (Schiffman
et al., 1995

). In addition to the
least squares mean perceived sweetness of each triad tested,
Table 3 also gives the mean responses of the associated self-mixtures
and the percentage difference of the mean response of each triad
from the average of the mean responses of its constituent self-mixtures.
The mean responses for the self-mixtures were based on experimental
measurements, because the doseresponse curves for sweeteners
are generally nonlinear (DuBois
et al., 1991

). For comparison,
Table 3 gives the average of the mean responses of the associated
dyads [found in Schiffman
et al. (1995)

], the average of the
mean responses of the constituent dyadic self-mixtures, as well
as the percentage difference of the mean responses of the dyads
from the average of their self-mixtures.
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Table 3 Comparison of ternary and binary mixtures (percentage difference of the mean response of each triad from the average of the mean responses of its constituent self-mixtures versus the percentage difference of the average response of the associated dyadsa from the average response of their self-mixtures)
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The following is an example of how the values in
Table 3 were
calculated. The triad aceapmsor
consists of acesulfame-K, aspartame and sorbitol. The first
value in the corresponding row of
Table 38.96is
the mean perceived sweetness of this triadic mixture. The second
value6.95 is the average of the mean perceived
sweetness of the three associated self-mixtures. The acesulfame-K
self-mixture is 5.33, for aspartame it is 5.45 and for sorbitol
it is 10.06. The third value28.9%is the percentage
difference between the self-mixtures average and the
mean response for the triad. This was calculated by subtracting
6.95 from 8.96 (2.01), then dividing this by 6.95, and multiplying
the result by 100 to give the percentage. The fourth value7.70is
the average of the mean responses of the associated dyads [from
Schiffman
et al. (1995)

]. The mean response of the dyadic mixture
aceapm is 7.86, for acesor it is 8.01 and for
apmsor it is 7.22. The fifth value5.96is
the average of the mean responses of the associated dyadic self-mixtures.
The acesulfame-K dyadic self-mixtures mean response is
4.75, for aspartame it is 4.70 and for sorbitol it is 8.43.
The sixth value29.2%is the percentage difference
between the self-mixtures average and the average of
the mean responses for the dyadic mixtures. This was calculated
by subtracting 5.96 from 7.70 (1.74), then dividing this by
5.96, and multiplying the result by 100 to give the percentage.
Results from other taste factors including temporal aspects, will be dealt with in a future paper.
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Discussion
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No clear trends to predict which triads are synergistic were
found, although the presence of a bulk sweetener (e.g. fructose,
glucose) tended to reduce synergism. This conclusion is consistent
with previous reports on synergism and taste modulation of sweeteners
(Portmann and Kilcast, 1996

; Hutteau
et al., 1998

; Birch, 1999

).
While the majority of sweetener triads tested in the present
study were found to be synergistic, it appears that a greater
degree of synergism may be reached using binary combinations
of sweeteners. When comparing the percentage increase in sweetness
intensity ratings of dyads with that of their constituent self-mixtures,
and triads with that of their constituent self-mixtures, it
was found that the percentage increase of the majority of the
triads is about the same or lower than that of the dyads. In
53 cases, the dyads had a greater degree of synergy than their
associated triads, and in only 25 cases was the reverse true.
It is possible that the greatest benefit from the synergism
of sweeteners is derived from the simple combination of two
sweeteners and not multiple sweeteners.
Further research is required fully to understand the chemical and biochemical mechanisms that produce synergy among specific sweeteners. This will involve a more complete understanding of transduction of sweet taste at the receptor level, the state of sweeteners in aqueous solution and quantitative structureactivity studies using molecular graphics and molecular dynamics simulations (Birch, 1996
).

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Figures 114 Figure 2 The least squares mean sweetness intensity ratings of every ternary combination tested containing acesulfame-K, alitame, aspartame, fructose, glucose, mannitol, Na cyclamate, Na saccharin, neohesperidin dihydrochalcone, rebaudioside-A, sorbitol, stevioside, sucrose and thaumatin, respectively. A dashed line is given to indicate a nominal response (i.e. an additive response).
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Figures 114 Figure 3 The least squares mean sweetness intensity ratings of every ternary combination tested containing acesulfame-K, alitame, aspartame, fructose, glucose, mannitol, Na cyclamate, Na saccharin, neohesperidin dihydrochalcone, rebaudioside-A, sorbitol, stevioside, sucrose and thaumatin, respectively. A dashed line is given to indicate a nominal response (i.e. an additive response).
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Figures 114 Figure 4 The least squares mean sweetness intensity ratings of every ternary combination tested containing acesulfame-K, alitame, aspartame, fructose, glucose, mannitol, Na cyclamate, Na saccharin, neohesperidin dihydrochalcone, rebaudioside-A, sorbitol, stevioside, sucrose and thaumatin, respectively. A dashed line is given to indicate a nominal response (i.e. an additive response).
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Figures 114 Figure 5 The least squares mean sweetness intensity ratings of every ternary combination tested containing acesulfame-K, alitame, aspartame, fructose, glucose, mannitol, Na cyclamate, Na saccharin, neohesperidin dihydrochalcone, rebaudioside-A, sorbitol, stevioside, sucrose and thaumatin, respectively. A dashed line is given to indicate a nominal response (i.e. an additive response).
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Figures 114 Figure 6 The least squares mean sweetness intensity ratings of every ternary combination tested containing acesulfame-K, alitame, aspartame, fructose, glucose, mannitol, Na cyclamate, Na saccharin, neohesperidin dihydrochalcone, rebaudioside-A, sorbitol, stevioside, sucrose and thaumatin, respectively. A dashed line is given to indicate a nominal response (i.e. an additive response).
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Figures 114 Figure 7 The least squares mean sweetness intensity ratings of every ternary combination tested containing acesulfame-K, alitame, aspartame, fructose, glucose, mannitol, Na cyclamate, Na saccharin, neohesperidin dihydrochalcone, rebaudioside-A, sorbitol, stevioside, sucrose and thaumatin, respectively. A dashed line is given to indicate a nominal response (i.e. an additive response).
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Figures 114 Figure 8 The least squares mean sweetness intensity ratings of every ternary combination tested containing acesulfame-K, alitame, aspartame, fructose, glucose, mannitol, Na cyclamate, Na saccharin, neohesperidin dihydrochalcone, rebaudioside-A, sorbitol, stevioside, sucrose and thaumatin, respectively. A dashed line is given to indicate a nominal response (i.e. an additive response).
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Figures 114 Figure 9 The least squares mean sweetness intensity ratings of every ternary combination tested containing acesulfame-K, alitame, aspartame, fructose, glucose, mannitol, Na cyclamate, Na saccharin, neohesperidin dihydrochalcone, rebaudioside-A, sorbitol, stevioside, sucrose and thaumatin, respectively. A dashed line is given to indicate a nominal response (i.e. an additive response).
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Figures 114 Figure 10 The least squares mean sweetness intensity ratings of every ternary combination tested containing acesulfame-K, alitame, aspartame, fructose, glucose, mannitol, Na cyclamate, Na saccharin, neohesperidin dihydrochalcone, rebaudioside-A, sorbitol, stevioside, sucrose and thaumatin, respectively. A dashed line is given to indicate a nominal response (i.e. an additive response).
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Figures 114 Figure 11 The least squares mean sweetness intensity ratings of every ternary combination tested containing acesulfame-K, alitame, aspartame, fructose, glucose, mannitol, Na cyclamate, Na saccharin, neohesperidin dihydrochalcone, rebaudioside-A, sorbitol, stevioside, sucrose and thaumatin, respectively. A dashed line is given to indicate a nominal response (i.e. an additive response).
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Figures 114 Figure 12 The least squares mean sweetness intensity ratings of every ternary combination tested containing acesulfame-K, alitame, aspartame, fructose, glucose, mannitol, Na cyclamate, Na saccharin, neohesperidin dihydrochalcone, rebaudioside-A, sorbitol, stevioside, sucrose and thaumatin, respectively. A dashed line is given to indicate a nominal response (i.e. an additive response).
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Figures 114 Figure 13 The least squares mean sweetness intensity ratings of every ternary combination tested containing acesulfame-K, alitame, aspartame, fructose, glucose, mannitol, Na cyclamate, Na saccharin, neohesperidin dihydrochalcone, rebaudioside-A, sorbitol, stevioside, sucrose and thaumatin, respectively. A dashed line is given to indicate a nominal response (i.e. an additive response).
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References
|
|---|
Ayya, N. and
Lawless, H. T. (1992)
Quantitative and qualitative evaluation of high-intensity sweeteners and sweetener mixtures. Chem. Senses, 17, 245259.
[Abstract/Free Full Text]
Bartoshuk, L.M. (1975) Taste mixtures: is mixture suppression related to compression? Physiol. Behav., 14, 642649.
Bartoshuk, L.M. and Cleveland, C.T. (1977) Mixtures of substances with similar tastes: a test of a psychophysical model of taste mixtures interactions. Sens. Proc., 1, 177186.
Birch, G.G. (1996) Towards an improved understanding of sweetener synergy. Trends Food Sci. Technol., 7, 401407.
Birch, G.G. (1999) Modulation of sweet taste. Biofactors, 9, 7380.[Web of Science][Medline]
DuBois, G.E., Walters, D.E., Schiffman, S.S., Warwick, Z.S., Booth, B.J., Pecore, S.D., Gibes, K., Carr, B.T. and Brands, L. (1991) Concentrationresponse relationships of sweeteners: a systematic approach. In Walters, D.E., Orthoefer, F.T. and DuBois, G.E. (eds), Sweeteners: Discovery, Molecular Design and Chemoreception, ACS Symposium Series 450. American Chemical Society, Washington, DC, pp. 261276.
Frank, R.A., Mize, S.J.S. and Carter, R. (1989) An assessment of binary mixture interactions for nine sweeteners. Chem Senses, 14, 621632.[Abstract/Free Full Text]
Hutteau, F., Mathlouthi, M., Portmann, M.O., and Kilcast, D. (1998) Physicochemical and psychophysical characteristics of binary mixtures of bulk and intense sweeteners. Food Chem., 63, 916.
Lawless, H.T. (1998) Theoretical note: tests of synergy in sweetener mixtures. Chem Senses, 23, 447451.[Abstract/Free Full Text]
Portmann, M.O. and Kilcast, D. (1996) Psychophysical characterization of new sweeteners of commercial importance for the EC food industry. Food Chem., 56, 291302.
Schiffman, S.S., Lindley, M.G., Clark, T.B. and Makino, C. (1981) Molecular mechanism of sweet taste: Relationship of hydrogen bonding to taste sensitivity for both young and elderly. Neurobiol Aging, 2, 173185.[Web of Science][Medline]
Schiffman, S.S., Pecore, S.D., Booth, B.J., Losee, M.L., Carr, B.T., Sattely-Miller, E.; Graham, B.G. and Warwick, Z.S. (1994) Adaptation of sweeteners in water and in tannic acid solutions. Physiol. Behav., 55, 547559.[Medline]
Schiffman, S.S., Booth, B.J., Carr, B.T., Losee, M.L., Sattely-Miller, E.A. and Graham, B.G. (1995) Investigation of synergism in binary mixtures of sweeteners. Brain Res. Bull., 38, 105120.[Web of Science][Medline]
Accepted October 7, 1999

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