Journal of Experimental Psychology: Copyright 2005 by the American Psychological Association

Human Perception and Performance 0096-1523/05/$12.00 DOI: 10.1037/0096-1523.31.1.101

2005, Vol. 31, No. 1, 101–109

Psychophysical Isolation of the Modality Responsible for Detecting

Multimodal Stimuli: A Chemosensory Example
Hisanori Nagata, Pamela Dalton, Nadine Doolittle, and Paul A. S. Breslin
Monell Chemical Senses Center

Multiple sense modalities can be stimulated conjointly by a physically complex item, such as a predator,
and also by a physically solitary stimulus that acts on multiple receptor classes. As a prime example of
this latter group, l-menthol from mint stimulates taste, smell, and several somatosensory submodalities.
In 6 experiments that used a variety of psychometric techniques, the authors experimentally isolated the
modality by which l-menthol is detected in the upper airways (the nose and mouth). Interestingly,
absolute detection in both the nasal and oral cavities was based on olfaction and not stinging, cooling,
or taste. These experiments illustrate how the sensory modality responsible for detecting a multimodal
or multisensory stimulus can be psychophysically isolated.

The ability to simultaneously use multiple sensory inputs to
perceive the external environment is an adaptive characteristic of
most vertebrates. Many real world entities, such as foods, preda-
tors, and even conspecific animals, are physically complex stim-
uli— emitting sounds, reflecting light, and releasing chemicals into
the air. Physically simple stimuli, such as chemicals, may also
stimulate multiple sensory systems and subsystems. To wit, recent
evidence within the chemical senses suggests that traditional
bitter-tasting compounds activate somatosensory neurons (Finger
et al., 2003; Liu & Simon, 1998), thermal stimuli induce taste
sensations (Cruz & Green, 2000), astringent compounds (alum)
elicit taste sensations (Breslin, Gilmore, Beauchamp, & Green,
1993), and a large majority of olfactory stimuli elicit irritation in
the nose (Cometto-Muñiz & Cain, 1995) and possibly pharynx.
Under these circumstances, it is very difficult to know which sense
modality is responsible for detection of the stimulus or object,
especially when qualitative information is lacking, as is often the
case at absolute detection (Stevens, 1995, 1997). There are even
individual stimuli that will stimulate several sensory systems si-
multaneously, including, taste, smell, pain, tactile, and thermal
sensations. The principal flavor in mint, l-menthol, is a prototyp-
ical example of this type of stimulus.
That a single physical stimulus may have multimodal action is
a widely accepted idea. To this point, l-menthol is widely used as
both a chemical cooling agent and as a counterirritant, that is, an
analgesic, for intrinsic irritation (e.g., from oral and pharyngeal
epithelia and from muscle). In the upper airways, l-menthol can
elicit sensations using three different modalities: somatosensation
Hisanori Nagata, Pamela Dalton, Nadine Doolittle, and Paul A. S.
Breslin, Monell Chemical Senses Center, Philadelphia, Pennsylvania.
This research was supported by National Institutes of Health (NIH)
Grant RO1DC03704 to Pamela Dalton and NIH Grant DC02995 to Paul
A. S. Breslin. We thank Ines Rodriguez and Jeanmarie Diamond for their
assistance with laboratory work and Barry Green for his advice regarding
olfactory lateralization techniques.
Correspondence concerning this article should be addressed to Paul
A. S. Breslin, Monell Chemical Senses Center, 3500 Market Street, Phil-
adelphia, PA 19104. E-mail: breslin@monell.org
(cooling, irritation, tingling, etc.), olfaction (minty), and gustation
(bitter). Regarding menthol, complex sensory and perceptual in-
teractions take place within the chemesthetic modality (Cliff &
Green, 1993, 1996; Green, 1985), from the activation of submo-
dalities such as irritation (burning, prickling, stinging, itching, etc.)
to thermal effects (cooling, warming) and tactile effects (buzzing,
tingling, tickling, numbing, etc.). In addition to stimulating odor
and taste, menthol elicits stinging, numbing, touch, and a cooling
sensation, which can inhibit the perception of warmth (Green,
1986, 1992).
Humans experience l-menthol most commonly in the upper
airways in the form of lozenges, chewing gums, cigarettes, tooth-
pastes, and mouth washes. Humans can detect the presence of
l-menthol in both the nasal and oral cavities; however, the relative
sensitivity of each of these regions and the modality by which
absolute detection of menthol occurs has not been well established.
Thus, as a prototypical example of a compound that stimulates
multiple sensory modalities, menthol could be detected using any
of these sensory systems in either the nose or mouth.
To illustrate this point, we conducted six experiments to psy-
chophysically isolate the sensory modality responsible for the
absolute detection of l-menthol. In Experiment 1, absolute detec-
tion thresholds were measured in the nasal cavity for l-menthol in
polyethylene glycol (PEG) 200. Oral detection thresholds with
aqueous solutions were collected with and without nose clips to
determine the effect of retronasal olfaction on oral thresholds
(Experiments 2, 3A, and 3B). Volatile compounds do not flow
retronasally into the nasal cavity when the nares are pinched close.
Thus, when one’s nares are pinched in an oral detection threshold
paradigm, oral sensations can be effectively isolated from nasal
sensations. Because intranasal chemosensation can be based on
either olfactory or somatosensory detection, we took advantage of
the fact that an olfactory sensation cannot be localized, or even
lateralized in humans (Kobal, Van Toller, & Hummel, 1989), and
collected intranasal lateralization thresholds to identify whether
nasal detection is olfactory or somatosensory based (Experiments
4 and 5). Experiments 3A, 3B, 4, and 5 attempted to refine the
possible modalities responsible for the oral and nasal detection of
l-menthol. Specifically, we asked whether oral thresholds were

101
102 NAGATA, DALTON, DOOLITTLE, AND BRESLIN
based on the taste, the odor, or the chemesthetic sensations in the
oral cavity from l-menthol (Experiments 3A and 3B), and once
oral detection was determined to be actually retronasal in origin,
we asked whether nasal thresholds were based on odor or chem-
esthetic sensations in the nasal passages (Experiments 4 and 5).
General Experimental Design
Participants
All participants were paid volunteers who provided signed consent using
a form that was approved by the Office of Regulatory Affairs at the
University of Pennsylvania. Participants were instructed not to eat, drink,
chew gum, or smoke for at least 1 hr prior to testing.
Procedures: Absolute Detection Thresholds (Experiments
1, 2, 3, and 5)
Absolute detection thresholds were measured with the two alternative
forced-choice method described by Stevens (1995). On each trial, the
participant was presented with both a stimulus and a blank in a random
order. After sampling the two stimuli (cups or bottle pairs) sequentially, the
participant was asked to identify which sample (first or second) contained
the stimulus. An incorrect response on any trial resulted in presentation of
the next higher concentration, whereas four consecutive correct responses
at a given concentration resulted in the presentation of the next lower
concentration (four-down, one-up staircase criteria). Testing continued
until five reversals (a correct decision followed by an incorrect decision or
vice versa) were collected. If, at any time, a participant moved more than
two consecutive concentration steps either up or down from the concen-
tration at which the first reversal occurred, then all reversals up to that point
were disregarded and five more reversals were required. These constraints
were used in order to clamp the acceptable concentration range of reversals
to within an order of magnitude. If the participant achieved five reversals
but did not meet the additional criteria, testing was continued until the
criteria were met. The threshold was calculated from the mean of the last
four reversals. Testing could be terminated under four different conditions:
(a) five valid reversals were achieved and the extra criteria were met,
resulting in a threshold value; (b) a participant failed to correctly detect the
stimulus at the highest concentration; (c) the participant correctly identified
the stimulus on four consecutive trials at the lowest concentration; (d) the
participant did not meet the criteria for a threshold before the 40th trial,
whereupon the session was terminated to prevent fatigue. Participants were
tested without trial feedback to focus their attention on their chemosensa-
tions, rather than teach them to attend to any correlated extraneous cues
(which we attempted to minimize; see below for details).
Data Analysis
Preliminary evaluation of the data indicated that the threshold ranges
were distributed lognormally. Consequently, the threshold concentration
values were transformed to logarithms before statistical analysis. Test–
retest reliabilities for nasal detection thresholds and oral detection thresh-
olds were assessed by Pearson’s product–moment correlation coefficients
and two-tailed t tests. The difference between the oral detection thresholds
with and without nose clips was evaluated using a t test for paired samples.
A p value was selected for two-tailed significance. For all tests of signif-
icance, the alpha error level was set at p ⫽ .05. All data analysis was
analyzed by SPSS software (SPSS, Inc., Chicago, IL).
Gas Chromatography
We used headspace analysis using gas chromatography to quantify the
amount of l-menthol in the headspace of both the nasal (in PEG200) and
oral (in deionized Millipore filtered water) stimuli. This was to determine
whether the concentrations of volatile l-menthol were similar for the two
methods and thereby link them using a common physical basis for detec-
tion, which in turn implicates the modality of detection. Vapor concentra-
tion of l-menthol in both solvents was measured using a Hewlett-Packard
6890 gas chromatograph equipped with a flame ionization detector linked
with a Hewlett-Packard Autosampler (HP7694). Splitless samples were
injected onto a SUPELCOWAX 10 capillary column (0.53-mm inner
diameter, 30-m long, film thickness ⫽ 0.50 ␮m) at a temperature program
that ramped from 100° C (1 min) to 180° C (10° C/min) to 220° C (25°
C/min), and detection occurred at 250° C. A four-point calibration curve
for l-menthol was generated by injecting known amounts of l-menthol
(10.0, 31.6, 100.0, and 316.0 ppm); their associated chromatographic areas
were used to convert the areas into concentration units (ppm ⫻ volume).
The coefficient of determination (r2) for the calibration curve was greater
than .98.
Experiment 1: Nasal Detection Thresholds
To our knowledge, there are no studies directly comparing the
sensitivity to l-menthol in the nose and in the mouth. Therefore,
baseline data must be collected to determine the absolute detection
thresholds for menthol in the two main upper airway passages.
Experiment 1 measured nasal detection thresholds in 100 volun-
teers in duplicate, and Experiment 2 measured oral detection
thresholds to menthol in triplicate. Experiment 1 involved an order
of magnitude 10 times more participants than did Experiment 2
because it also generated data to help determine whether the
frequency histogram for sensitivity to l-menthol would deviate
from unimodality, possibly indicating a small number of mecha-
nisms involved in detection, as is seen with distributions that
appear bimodal.
Method
Participants. One hundred volunteers were paid to participate after
signing an Institutional Review Board approved consent form (as was true
in all subsequent experiments below). Participants ranged in age from 16
to 69 years (average age ⫽ 33.5 years), with equal representation of males
and females. Fifteen participants had previous experience in psychophys-
ical experiments.
Stimuli. The nasal series included 26 samples ranging in concentration
from 1.56 ⫻10⫺9 M to 0.1016 M l-menthol in PEG200 in one quarter log
steps. A total of six blanks were used (PEG200 only), and new blanks were
prepared daily. Nasal threshold series were made on Monday and used for
1 week only. Stimuli were presented to participants in 280-ml glass bottles
fitted with specially designed Teflon nose pieces. Glass bottles contained
either 10 ml of a solution of l-menthol in PEG200 solution (odorant) or the
same amount of PEG200 (blank). The bottles were wrapped in thin foam
material to help eliminate any external temperature influences (bottles were
maintained at a room temperature of approximately 21° C) and to thermally
insulate the bottles when handled.
Procedure. Participants and experimenters handled bottles while wear-
ing polypropylene gloves to further reduce transmission of extraneous cues
in the detection task. On each trial, the participant was presented with two
pairs of glass bottles. One pair contained l-menthol and a blank (odorant
pair); the other pair contained two blanks only (blank pair). The partici-
pants simultaneously sniffed from both bottles within a pair and sniffed
from both pairs sequentially, then decided which pair of bottles (first or
second) contained the odorant. The l-menthol was placed randomly on the
left or right bottle within the odorant pair and was presented randomly
either in the first or second set of bottles. To avoid adaptation and/or
sensitization, we used a 30-s intertrial interval (ITI). We selected this
 PSYCHOPHYSICAL ISOLATION OF SENSORY MODALITY
interval because pilot testing indicated that 30 to 45 s minimized both
sensitization and desensitization to irritation over several suprathreshold
l-menthol exposures in 6 participants. The pilot test requested that they
sample 10 menthol solutions at a fixed intersolution interval ranging from
10 s to 5 min and rate irritation from l-menthol after each sampling.
Different ITIs were tested on different days. The 30-s ITI was extended
slightly if a participant required more than 30 s to complete a trial. The test
began with Bottle Number 3 (0.0065 M). Each participant completed two
nasal detection threshold sessions with a 15-min break between the two
sessions. The participant left the test room during the break.
Results and Discussion
Individual threshold concentrations varied from 5.00 ⫻ 10⫺5 M
to 5.10 ⫻ 10⫺2 M (three orders of magnitude variation) l-menthol
in PEG200 (relative headspace concentration ranging from 0.002
␮g/L to 11.600 ␮g/L), with an exception of two data points
(3.00 ⫻ 10⫺3 M, 1.20 ⫻ 10⫺2 M), and with an overall geometric
mean threshold concentration of 3.42 mM (see Figure 1). The
frequency distribution of nasal thresholds was unimodal, appeared
approximately normal, and was found to be more consistent and to
fall within a tighter range than has been previously reported for the
nasal thresholds of many other olfactory compounds. Interestingly,
among the 100 participants tested, there were no significant rela-
tionships between nasal sensitivity to l-menthol and age, gender, or
smoking status. Test–retest reliability was quite good (r ⫽ .79, p ⬍
.01; see Figure 2). The slope deviates slightly from unity because
a few participants had much lower thresholds on their second test.
Experiment 2: Oral Detection Thresholds With the Nose
Open
Method
Participants. Ten volunteers (4 women and 6 men) were paid to
participate. Participants ranged in age from 18 to 32 years (average age ⫽
Figure 1. A frequency histogram of 200 l-menthol nasal detection thresh-
olds (two thresholds from 100 participants) from Experiment 1. The y-axis
depicts the number of participants in each threshold category bin. The
x-axis depicts the various millimolar concentration bin categories of
l-menthol in polyethylene glycol (PEG) 200. The distribution is approxi-
mately lognormal, spanning three orders of magnitude, and is unimodal in
form. The geometric mean nasal threshold level was 3.42 mM.
103
Figure 2. The test–retest reliability of l-menthol nasal detection thresh-
olds for 100 participants in Experiment 1. The x-axis represents the
threshold value of each participant from Test 1 in log molar. The y-axis
represents the threshold value of each participant from Test 2 in log molar.
The test–retest reliability was highly significant. The slope departs from
unity because a few participants had much lower thresholds on Test 2.
23.5 years). All participants had previous experience in psychophysical
experiments.
Stimuli. The oral threshold series contained 13 samples, ranging in
concentration from 4.80 ⫻ 10⫺7 M to 4.75 ⫻ 10⫺4 M l-menthol in
deionized Millipore filtered water in one quarter log steps. Deionized
Millipore filtered water was used for blanks as well as for rinsing between
samples. All stimuli and blanks were maintained at a constant temperature
by immersion of bottles in a circulating water bath (Cole-Parmer Polystat)
held at 25° C. Fresh oral threshold series were prepared weekly.
Procedure. On each trial, participants were given two samples (10 ml
each) in 40-ml plastic medicine cups (Baxter). They were instructed to take
the first sample, hold it in their mouth for 5 s, expectorate, and rinse.
Participants then repeated this procedure with the second sample and
decided which sample contained the stimulus. Participants breathed natu-
rally through the nose while resting between trials. Cups were presented in
random order. A 45-s ITI was used for oral thresholds to minimize
sensitization and desensitization effects. Three thresholds were collected
on different days.
Results and Discussion
Averaged oral threshold concentration for each participant
ranged from 1.40 ⫻10⫺6 M to 1.43 ⫻10⫺4 M l-menthol in water
with an overall geometric mean threshold concentration of 9.00 ⫻
10⫺6 M (see Figure 3). The distribution of oral detection thresh-
olds was also unimodal and spanned more than two orders of
magnitude. Test–retest reliability was good and increased slightly
between Test 1-Test 2 (r ⫽ .61, p ⬍ .01) and Test 2-Test 3 (r ⫽
.67, p ⫽ .17; see Figure 4).
Experiment 3A: Oral Detection Thresholds With the Nose
Closed or Open
Experiments 1 and 2 allow one to compare nasal and oral
absolute detection thresholds with l-menthol and revealed certain
similarities. For example, both frequency-histogram distributions
were unimodal and spanned approximately two to three orders of
magnitude (compare Figures 1 and 3). These data do not, however,
indicate the sensory modality by which menthol was detected.
Thus, oral menthol detection could be based on taste, odor, or
somatosensory sensations. Experiment 3 compared oral threshold
104 NAGATA, DALTON, DOOLITTLE, AND BRESLIN

testing when the nares were open or when they had been closed
with nose clips. There can be no retronasal flow of volatile
compounds to the nasal cavity from the mouth when the nares are
pinched closed. Therefore, this simple technique serves to isolate
the two upper respiratory passageways. If the detection thresholds
are the same, then detection of menthol is oral and olfaction can be
ruled out. If the oral detection threshold is higher when the nares
are closed, then retronasal flow of volatiles will appear critical for
oral detection.

Figure 3. A frequency histogram of 29 l-menthol oral detection thresh-
olds (three thresholds from 10 participants; 1 participant was unable to test
a third time) from Experiment 2. The y-axis depicts the number of partic-
ipants in each threshold category bin. The x-axis depicts the various
micromolar concentration bin categories of l-menthol in deionized Milli-
pore filtered water. The distribution is approximately unimodal in form and
spans more than two orders of magnitude. The geometric mean oral
threshold level was 9.0 ⫻10⫺6 M.
Method
Participants. Participants were the same as in Experiment 2.
Stimuli. Samples were the same as in Experiment 2.
Procedure. To determine the effect of retronasal olfaction on the oral
threshold for l-menthol, we compared the oral threshold for l-menthol in
water when the retronasal flow of l-menthol vapor was permitted and the
oral threshold for l-menthol when retronasal flow was blocked by wearing
plastic and foam rubber nose clips (Diagnostic Medical System). In the first
condition, the participants were instructed to put on the nose clips, rinse,
take the first sample, hold for 5 s, and then expectorate. The participants
would follow the same procedure with the second sample, followed by a
judgment of which sample contained the stimulus. The participants could
then remove the nose clips until the next set of samples was presented.
Participants were not instructed how to breathe during the ITI.
In the second condition, thresholds were obtained without nose clips.
Participants were instructed to rinse, take the first sample, hold for 5 s

Figure 4. The test–retest reliability of l-menthol oral detection thresholds for the 9 participants in Experiment
2 who completed all three oral thresholds. All axes depict the oral threshold concentration in log millimolar
l-menthol. Test 1 to Test 2 reliability is depicted in the top panel, Test 2 to Test 3 reliability is in the lower left
panel, and Test 1 to Test 3 reliability is in the lower right panel.
 PSYCHOPHYSICAL ISOLATION OF SENSORY MODALITY 105

while breathing through the nose, and then expectorate. After following the Table 2
same procedure for the second sample, they were asked to judge which cup Individual Geometric Means of Oral Threshold With and
contained the stimulus. Because the purpose of this study was to determine Without Nose Clips in Experiment 3B
the role of retronasal olfaction on the detection of an orally presented
stimulus, we wanted to maintain retronasal olfaction as sensitive as pos- Participant mM
sible. Therefore, during the ITI, participants were instructed to breathe only
through the mouth to avoid any possible adaptation of olfaction and/or Without nose clips
pharyngeal irritation from residual l-menthol vapor in the upper airways or 1 0.1240
in the ambient room air. Participants were tested in a total of three sessions. 2 0.1182
3 0.0604
Within a single session, each participant was tested in both conditions with
4 0.0089
a 15-min break between conditions. Order of conditions was counterbal- 5 0.1126
anced across sessions and, as always, stimulus order was randomized. 6 0.0604
7 0.1576
Results and Discussion 8 0.1909
9 0.0130
The results of Experiment 3A are shown in Table 1; the results 10 0.0523
of Experiment 3B (discussed below) are shown in Table 2. Thresh- Geometric M 0.0636
olds for oral l-menthol obtained without retronasal flow (wearing With nose clips
nose clips) were significantly greater ( p ⬍ .01), by an order of
magnitude, than thresholds for oral l-menthol obtained when ret- 1 0.2205
ronasal flow was unimpeded (7.46 ⫻ 10⫺5 M vs. 7.70 ⫻ 10⫺6 M; 2 0.2546
3 0.2426
see Table 1). These oral thresholds both with and without nose 4 0.0499
clips show little relationship (r2 ⫽ .11; negative trend) and, in light 5 0.2802
of our high test–retest reliability (see Figures 2 and 4), suggest that 6 0.1126
these thresholds are based on independent mechanisms. 7 0.0767
The headspace concentration of samples used in Experiments 1, 8 0.2940
9 0.0731
2, and 3 were analyzed with gas chromatography. Figure 5 shows 10 0.0548
Geometric M 0.1345

Table 1 Note. This table presents the results of Experiment 3B and parallels Table
Individual Geometric Means of Oral Threshold With and 1. The top section presents oral thresholds with nose clips on during
Without Nose Clips in Experiment 3A delivery followed by nose clip removal during assessment, enabling purely
retronasal olfaction, and the bottom section is the same as in Table 1.
Participant mM

Without nose clips
1 0.0103
2 0.0056
3 0.0119
4 0.0039
5 0.0075
6 0.0087
7 0.0077
8 0.0178
9 0.0016
10 0.0189
Geometric M 0.0077
With nose clips
1 0.2450
2 0.0820
3 0.0291
4 0.0975
5 0.1125
6 0.0615
7 0.0581
8 0.0388
9 0.1263
10 0.0475
Geometric M 0.0746
Note. The geometric mean results of Experiment 3A for each participant
comparing oral thresholds without nose clips, enabling both orthonasal and
retronasal olfaction (top section) and oral thresholds with nose clips,
disabling olfaction (bottom section). For comparison, the geometric mean
oral threshold from Experiment 2 is .009 mM.
the average headspace concentration of each sample. Although
measurements could not be made near the concentration of oral
thresholds, we calculated the headspace concentration at oral
threshold by extrapolating Henry’s law (P ⫽ x ⫻ K) to these
concentrations (where P is vapor pressure, x is molar fraction, and
K is a constant).
Although the liquid concentrations differed between nasal
thresholds (3.42 ⫻10⫺3 M in PEG200 [Experiment 1]) and oral
thresholds (9.00 ⫻ 10⫺6 M in water [Experiment 2]) by almost
three orders of magnitude, the headspace concentrations were
approximately the same (0.43 ␮g/L and 0.15 ␮g/L, respectively;
see Figure 5). The headspace concentration of the oral thresholds
was slightly lower than the headspace concentration of nasal
thresholds. There are three main possibilities that account for this
small difference. One possibility is that when samples were held in
the mouth, the liquid temperature increased slightly because of the
increased temperature of the mouth, relative to room temperature.
If the liquid temperature increased, the headspace concentration
would increase in proportion to liquid temperature. When 10 ml of
21° C water is introduced to the oral cavity for 30 s, the temper-
ature increases to 32° C, as measured by a digital thermometer in
our laboratory. When aqueous menthol was heated to this temper-
ature, the headspace concentration was even closer to the nasal
threshold bottles’ headspace concentration. A second possibility is
that detection in the oral cavity was facilitated by the integration of
odor and taste. At suprathreshold levels, integration of the two
106 NAGATA, DALTON, DOOLITTLE, AND BRESLIN

Figure 5. The headspace concentrations of nasal (left panel) and oral (right panel) l-menthol thresholds against
the concentrations of l-menthol in liquid vehicle samples. In the left panel, the x-axis depicts the log molar
concentration of l-menthol in polyethylene glycol (PEG) 200, and in the right panel the x-axis depicts the log
millimolar concentration in deionized Millipore filtered water. In both panels, the y-axis represents the log
headspace concentration in ␮g/L. The vertical dashed line shows the geometric mean l-menthol thresholds
(3.420 mM, left panel; 0.009 mM, right panel). The horizontal dashed line shows the headspace concentration
of the threshold samples (0.43 ␮g/L, left panel; 0.15 ␮g/L, right panel).

modalities has been reported (Gillan, 1983; Murphy & Cain,
1979). Previous work in our laboratory has provided evidence in
humans of subthreshold, cross-modal chemosensory integration
(Dalton, Doolittle, Nagata, & Breslin, 2000). When l-menthol
samples are in the oral cavity, the l-menthol headspace concentra-
tion may be slightly below the retronasal threshold concentration,
but integration between the retronasal odor and the taste of men-
thol could enable the increased absolute sensitivity. And third, the
orthonasal and retronasal olfactory thresholds may simply differ
slightly because of the exposure to different epithelial surfaces
from the two directions of flow in the airway. Detection of
l-menthol using oral thresholds occurred at much lower concen-
trations when thresholds were measured without nose clips, thus
implicating retronasal sensory cues as an important determinant of
the sensitivity to l-menthol when presented orally.
Experiment 3B: Oral Detection Thresholds With the Nose
Closed or Open for Retronasal Olfaction
In Experiment 3A, the nose-open condition allowed retronasal
olfaction to occur, which is a major component of oral-based
olfaction and food flavor. We assumed that the majority of olfac-
tory input from oral stimuli was using this pharyngeal pathway.
We did not, however, explicitly eliminate orthonasal input. As the
menthol solution was drawn into the oral cavity, it must first pass
under the open nose. This creates an opportunity for brief but
possibility sufficient sampling of volatiles for detection as the oral
sample is taken. Our observations indicate that, under most in-
stances, slight nasal inspiration of air accompanies the solution
being drawn into the oral cavity. Thus, volatiles may be sampled
even as the liquid bolus passes the lips. Therefore, we conducted
Experiment 3B expressly to mirror Experiment 3A but to eliminate
orthonasal input and to compare detection of menthol without any
olfaction to detection with only retronasal olfaction.
Method
Participants. Ten naive volunteers (5 women) were paid to participate.
Participants ranged in age from 19 to 36 years (average age ⫽ 23.7 years).
Seven participants had previous experience in psychophysical experiments
but had not participated in these menthol studies.
Stimuli. Samples were the same as in Experiment 2.
Procedure. To determine the effect of retronasal olfaction on oral
thresholds for l-menthol, we compared thresholds under two conditions. In
one condition, retronasal and orthonasal flow was blocked; in the second,
retronasal flow was permitted only while the stimulus was in the mouth. In
the first condition, participants were given two samples (10 mL each) in
40-mL plastic medicine cups. They were instructed to hold the nares closed
with foam rubber nose clips, rinse, take the first sample, hold it in their
mouth for 5 s, expectorate, and rinse. They repeated the procedure with the
second sample and judged which sample contained the stimulus. Cups were
presented in random order. A 45-s ITI was exercised, during which the
nose clips were removed and participants were allowed to breathe
normally.
In the second condition, participants were given two samples (10 mL
each) in 40-mL plastic medicine cups. They were instructed to hold the
nares closed with foam rubber nose clips, rinse, take the first sample into
the mouth, remove the nose clips, and hold it in their mouth for 5 s while
breathing through the nose. After holding the sample in the mouth for 5 s,
participants were instructed to replace the nose clips, expectorate, and
rinse. They repeated the procedure with the second sample and judged
which sample contained the stimulus. L-menthol and blank cups were
presented in random order. A 45-s ITI was exercised, during which the
nose clips were removed. Participants were also instructed to breathe
normally during this period. On each of 3 days, participants were tested
once in each condition with a 15-min break between conditions. Conditions
were counterbalanced among participants and visits.
Results and Discussion
Thresholds for oral l-menthol obtained with the nares closed
(1.35 ⫻10⫺4 M) were significantly greater ( p ⬍ .01) than thresh-
 PSYCHOPHYSICAL ISOLATION OF SENSORY MODALITY
olds obtained with retronasal flow permitted (6.36 ⫻10⫺5 M; see
Table 2). Thus, these results confirm the general conclusion of
Experiment 3A, that the detection of l-menthol in the oral cavity is
based on volatiles entering the nose rather than orally derived
sensory input. Although Experiment 3A permitted both orthonasal
and retronasal olfaction in the nose-open condition, the present
experiment permitted only retronasal olfaction during oral sam-
pling. As in Experiment 3A, the oral thresholds in Experiment 3B
both without olfaction and with retronasal olfaction showed little
relationship (r2 ⫽ .38; positive trend). This weak trend is in the
opposite direction of the trend seen in Experiment 3A. Taken
together these two experiments strongly suggest that both of these
types of oral l-menthol thresholds (nose open and closed) are based
on independent mechanisms.
The difference in sensitivity between the nose-closed and the
nose-open conditions in Experiment 3A was almost tenfold,
whereas the difference between the two conditions in Experiment
3B was slightly more than twofold. Therefore, it appears that
olfactory detection is more sensitive when utilizing both orthona-
sal and retronasal pathways than when only retronasal pathways
are engaged. Because different participants were used in Experi-
ments 3A and 3B and they differed slightly in their sensitivities to
l-menthol with the nares closed (Experiment 3B participants were
1.75 times less sensitive), it is difficult to draw absolute conclu-
sions regarding the relative roles of orthonasal olfaction and ret-
ronasal olfaction when sampling a solution orally. It is possible
that orthonasal and retronasal inputs combine additively to in-
crease olfactory signal strength, but the present studies do not
permit this conclusion. Regardless, because Experiment 3B par-
ticipants were 4.5 times less sensitive than the Experiment 3A
participants in the nose-open condition relative to differences in
their nose-closed sensitivities, we can conclude that orthonasal
sampling of volatiles when drawing a solution into the oral cavity
also plays a role in detecting oral l-menthol solutions.
Experiment 4: Nasal Lateralization Threshold
In the nasal cavity, l-menthol can elicit odor and chemesthetic
sensations; however, it is not clear by which modality absolute
detection in the nasal cavity is based. Experiments 3A and 3B
demonstrate that l-menthol detection is based on volatile menthol
entering the nose but do not identify whether nasal olfaction or
nasal somatosensation is involved in the detection. Nasal lateral-
ization has been a useful technique for measuring chemesthetic
thresholds in the presence of odor sensations (Cometto-Muñiz &
Cain, 1995; Wysocki, Dalton, Brody, & Lawley, 1987; Wysocki,
Green, & Malia, 1992). Evidently, only somatosensory signals can
be lateralized within the two nares. Thus, if the lateralization
threshold is much higher than the absolute nasal detection thresh-
old, then nasal detection can be deduced to be olfactory. If later-
alization thresholds in the nose are similar to absolute detection,
then nasal detection can be deduced to be somatosensory.
Method
Participants. Six adult volunteers (4 women and 2 men) were paid to
participate. Participants ranged in age from 18 to 32 years (average age ⫽
25.3 years). All participants had previous experience in psychophysical
experiments, and 4 participants had participated in Experiments 2 and 3.
107
Stimuli. An eight-bottle series ranging in concentration from 7.90 ⫻
10⫺4 M to 0.203 M in one quarter log steps was prepared in the same
manner as described in Experiment 1 by dissolving l-menthol in PEG200.
Procedure. On each trial, participants were presented with only one
pair of bottles. One bottle contained l-menthol in PEG200; the other bottle
contained only PEG200. The participants simultaneously sniffed the pair
and then decided which bottle (left or right) contained the stimulus.
Thresholds were collected using the method of fixed stimuli with an
ascending staircase beginning with the lowest concentration (7.90 ⫻ 10⫺4
M). Testing continued until five correct responses were reported at a given
concentration. This concentration was considered the lateralization thresh-
old concentration. Two ITIs (2 and 5 min) were again manipulated to test
for effects of adaptation and sensitization of the chemesthetic properties of
l-menthol. Each participant was tested twice with both ITIs.
Results and Discussion
In Experiment 3B, we observed retronasal olfaction to be more
sensitive than orally restricted sensitivity to l-menthol samples.
The question remained, however, whether the retronasal detection
of l-menthol was based on olfactory or chemesthetic sensations. In
this experiment, we measured chemesthetic thresholds in the nasal
cavity using the nasal lateralization technique (Wysocki et al.,
1987, 1992) to compare them with the nasal detection thresholds
obtained in Experiment 1.
The results are given in Table 3 for those who also participated
in Experiment 1. For all participants, the lateralization thresholds
(2-min and 5-min ITIs) were one to two orders of magnitude
higher than the nasal detection thresholds. The ITI was also varied
in the lateralization procedure in order to examine the effect of
repeated presentations of high concentrations of l-menthol on
chemesthetic sensitivity. During one test session, thresholds were
obtained with a 2-min ITI and during another session, they were
obtained with a 5-min ITI. Overall, the results showed no differ-
ence in sensitivity based on the different ITIs (8.00 ⫻ 10⫺2 M for
a 5-min ITI vs. 9.00 ⫻ 10⫺2 M for a 2-min ITI).
If we accept the premise that lateralization thresholds depend on
the participant’s ability to detect chemesthetic sensations in the
nasal passages and that pure odor signals cannot be localized in the
nose (Kobal et al., 1989), then it appears that nasal detection of
l-menthol (both orthonasal and retronasal) was based on odor
sensations and occurred at far lower concentrations than those
necessary to permit nasal lateralization.
Table 3
Geometric Means of L-Menthol in Polyethylene Glycol 200
Nasal lateralization
threshold
Nasal detection
Participant threshold 5-min ITI 2-min ITI
1 0.0016 0.0762 0.1365
2 0.0008 0.1524 0.0683
3 0.0010 0.1016 0.0254
4 0.0016 0.0190 0.1360
Note. This table compares geometric mean results of Experiment 4 for
nasal absolute detection thresholds and nasal lateralization thresholds for
each participant. ITI ⫽ intertrial interval.
108 NAGATA, DALTON, DOOLITTLE, AND BRESLIN
Experiment 5: Nasal Detection and Lateralization
Thresholds Measured Concurrently
It is difficult to directly compare the values obtained in olfactory
detection tasks with those obtained in lateralization tasks because
of the increased cognitive demands imposed by the task of odorant
lateralization. For example, in both tasks the individual is (explic-
itly or implicitly) asked to detect whether the stimulus is present.
However, in the lateralization task, the individual is also required
to indicate the location of the odorant (right or left nostril). Thus,
the higher thresholds that are typically obtained for odorant later-
alization could potentially result from the additional cognitive
demand imposed by this task. However, in a recent study that
obtained odor thresholds and lateralization thresholds in exactly
the same manner (i.e., asking participants to detect and localize the
odor–irritant on each trial), participants were able to detect odor at
much lower concentrations than they were able to accurately
lateralize it (Radil & Wysocki, 2000; Wysocki et al., 1992).
The method used in Experiment 5 was designed to minimize the
cognitive demand differences between the detection and lateral-
ization thresholds. The participant was asked the same question
throughout the entire study (Which bottle contains the stimulus?)
using a single threshold protocol regardless of the type of threshold
being collected.
Method
Participants. Eight adult volunteers (3 women and 5 men) ranging in
age from 20 to 33 years (average age ⫽ 25.6 years) were paid to
participate.
Stimuli. A nasal series of l-menthol in PEG200 was prepared using the
same method described for Experiment 1.
Procedure. Just as in the detection threshold measurement from Ex-
periment 1, participants were presented with two pairs of glass bottles on
each trial. One pair contained l-menthol and a blank; the other pair
contained two blanks only. The participants simultaneously sniffed from
both bottles within a pair and sniffed from both pairs sequentially, then
decided which individual bottle contained the odorant. Participants fol-
lowed this method throughout the entire experiment. As in Experiments
1–3, a modified staircase procedure with a four-down, one-up, five-reversal
criteria was used.
First, an absolute detection threshold was measured, followed by a
lateralization threshold. To collect the absolute detection thresholds, we
made movement up or down in concentration depending on whether the
participants correctly identified the pair of bottles containing the l-menthol
when they reported which individual bottle they believed contained the
stimulus. After five reversals were achieved for the detection threshold, the
lateralization testing would begin at the concentration of the detection
threshold. Movement, up or down in concentration, was then made de-
pending on whether the participant correctly identified the individual bottle
containing the l-menthol.
Results and Discussion
The results are very similar to the findings reported for Exper-
iments 1 and 4, for odor and lateralization thresholds, respectively.
The mean absolute detection threshold concentration in Experi-
ment 5 was 2.00 ⫻10⫺3 M (very similar to the Experiment 1
results), and the mean lateralization threshold concentration was
9.00 ⫻ 10⫺2 M (virtually identical to the Experiment 4 results).
For every participant tested, the lateralization threshold was higher
than the detection threshold concentration. It is very unlikely that
order effects are responsible for the difference seen here between
threshold types because responses were highly stable in Experi-
ment 1 (see Figure 2) when only a brief pause was inserted
between two threshold tests.
Although the threshold protocol used in Experiment 4 is much
simpler than that used in the other experiments, it nevertheless
resulted in lateralization values very similar to those obtained in
Experiment 5 using a modified staircase with two pairs of bottles
to identify both olfactory and chemesthetic thresholds concur-
rently. Therefore, Experiment 5 effectively validates the simpler
and more commonly used method of nasal lateralization by as-
cending staircase of fixed stimuli (Experiment 4).
General Discussion
The objective of this study was to illustrate how to psychophys-
ically isolate the modality by which a multimodal stimulus is
detected. Given that the detection and perception of most stimuli
involves multiple sensory modalities and/or submodalities, this
approach has broad application, especially for gustatory, olfactory,
and somatosensory chemical stimuli, as well as more traditional
somatosensory and visual stimuli. For this purpose, we selected a
model stimulus, l-menthol, which can act on three sensory modal-
ities but for which the basis of absolute detection was previously
unknown. The experiments reported here demonstrate that humans
detect l-menthol in the upper airways using olfaction and not by
taste or somatosensations, regardless of whether it is delivered
nasally or orally.
Frequency histograms of absolute detection thresholds for
l-menthol in the nose and the mouth were unimodally distributed.
The distributions of nasal and oral detection thresholds had high
test–retest reliability. The observation that menthol detection
thresholds are unimodally distributed in the sample population
suggests that sensitivity for menthol has multiple determinants. A
unimodal distribution is traditionally interpreted to mean that
multiple factors with additive effects create a continuum of vari-
ability within the population. Conversely, a bimodal or trimodal
distribution is often interpreted to mean that one or two variations
in a single factor, such as a l-menthol receptor, is separating the
population into two or three clusters as a function of the number of
variants. The variability of nasal (orthonasal) and oral (retronasal)
thresholds across participants spanned two to three orders of
magnitude. Thus, some participants may be 1,000 times more
sensitive than others at detecting l-menthol. The variability was
somewhat smaller when menthol was detected retronasally with
oral delivery, but this is likely due to low sampling. We measured
200 nasal thresholds from 100 participants and only 59 oral–
retronasal thresholds from 20 participants in Experiments 3A and
3B. Variability for oral thresholds may well have increased if 10
times more participants were tested.
When retronasal flow was blocked by nose clips (Experiments
3A and 3B), thresholds for oral l-menthol were significantly higher
in concentration than oral thresholds without nose clips, indicating
that olfaction can be an important sensory cue during testing of
sensitivity for orally presented l-menthol. This was true whether
olfactory input from the oral cavity was orthonasal and retronasal
or merely retronasal. When olfaction was restricted to the retro-
nasal pathway, however, sensitivity was lower than when both
olfactory pathways were permitted, which suggests that volatiles
 PSYCHOPHYSICAL ISOLATION OF SENSORY MODALITY
are sampled orthonasally from solutions as they are sipped and this
pathway increases olfactory sensitivity. The chemesthetic lateral-
ization thresholds from two methods (Experiments 4 and 5) were
higher in concentration than the nasal absolute detection thresh-
olds, suggesting that l-menthol can be detected in the nose at
concentrations that do not permit chemesthesis. Gas chromato-
graphic analysis indicates that both oral and orthonasal l-menthol
thresholds were very similar with respect to headspace concentra-
tion of l-menthol, differing by less than a factor of three (0.43 and
0.15 ␮g/L), whereas their solution concentrations differed by
almost three orders of magnitude. These results strengthen our
conclusion that the absolute detection of l-menthol, both nasally
and orally, was based on a volatile compound in the nose, rather
than l-menthol’s more poignant somatosensory qualities of sting-
ing and cooling in the mouth. The failure of lateralization threshold
concentrations to match absolute detection threshold concentra-
tions suggests that somatosensory stinging and cooling in the nose
was not the basis of detection either. Detection of l-menthol was
olfactory, regardless of the route of delivery.
Given that many stimuli act on more than one sensory modality,
it is often not obvious which sense subserves absolute detection.
For chemical stimuli, oral and nasal sensations can be isolated with
the use of nose clips to block all ortho- and retronasal flow of
volatiles. In the case of olfaction and nasal irritation, lateralization
is a powerful technique because pure olfactory stimuli are not
lateralizable (Kobal et al., 1989; Radil & Wysocki, 2000; Wysocki
et al., 1992). Oral–lingual stimuli, however, cannot be separated
with the same techniques, because gustatory stimuli are localizable
as a function of gustatory qualities without discriminative somato-
sensory cues (Delwiche, Lera, & Breslin, 2000; Shikata, Mc-
Mahon, & Breslin, 2000). In addition, it is difficult to deliver an
oral chemical stimulus without concomitant tactile stimulation in
the same areas. Nevertheless, l-menthol, as shown by this study,
serves as an excellent example of how the contribution of multiple
sensory modalities, stimulated by a single compound, can be
analytically studied and isolated using a combination of psycho-
physical techniques.
Although there are clear evolutionary benefits for sensory sys-
tems to operate in combination to perceive external stimuli through
multiple modalities and submodalities, the ability to decompose
the gestalt percept in order to identify the basis for stimulus
detectability is an important tool for (a) evaluating the degree of
additivity or synergy among sensory modalities and (b) examining
how the brain integrates inputs from different sensory systems
(Dalton et al., 2000). This process of psychophysical isolation of
sensory subsystems is also applicable to the perception of complex
stimuli that are not chemical in origin. Other examples of stimulus
detection in which the specific roles of subsystems are unclear
include visual identification– detection of an object, which could
involve shape, color, shading, stereoscopic cues, and so forth, and
the detection of cold water, which could include systems ranging
from touch, to cool, to polymodal nociceptors. Which subsystems
are used in the initial detection of the object–stimulus could be
studied using similar analytical approaches to the one demon-
strated in the present study with l-menthol. The strategies used in
the present study to psychophysically isolate and determine the
sensitivity of each modality contributing to the perception of a
multimodal stimulus can have broad applicability in multisensory
research.
109
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Received February 20, 2002
Revision received April 9, 2004
Accepted July 6, 2004 䡲