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Title: Factors Affecting the Ortho- and Retronasal Perception of Flavors: A Review

Authors: Erin M. Goldberg1+, Kun Wang2+, Jessica Goldberg1+ and Michel Aliani1, 3ffi

Department of Human Nutritional Sciences1, Food Science2, University of Manitoba; Canadian

Centre for Agri-Food Research in Health and Medicine3, St Boniface Research Centre,

Winnipeg, Manitoba, R2H 2A6

+
These authors made equal contribution to the preparation of this review paper.

ffi Corresponding author

Email: Michel.Aliani@Umanitoba.ca

Canadian Centre for Agri-Food Research in Health and Medicine

St. Boniface Hospital Research Centre, 351 Tache Ave, Winnipeg, MB, Canada, R2H 2A6

Abstract

Flavor perception is a highly individual sensation, and is impacted by a number of factors.

Olfaction is a critical element in fully experiencing flavor. In this review, we explore the

differences between orthonasal (sniff) versus retronasal (mouth) olfaction, and provide a

comprehensive summary of recent publications in this arena. Here we explore the complexities

of flavor perception, including the role that select flavors and media have on identification and

localization. We also discuss some common neural imaging techniques used in this field, as

odorants activate different neural responses in diverse areas of the brain, as well as the different

stimulation patterns derived from perceiving food and non-food related odorants. The

information provided will be useful for sensory scientists and industry alike for the development

of novel food and beverages that positively impact the consumer experience.

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Key words

Orthonasal, Retronasal, Olfaction, Brain Imaging, Flavor Perception

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1. Introduction

Flavor perception is a multi-factor experience that involves olfactory (aroma), gustatory

(taste), texture, trigeminal sensations, visual cues and the integration of all of these sensory

modalities (Gautam & Verhagen, 2010; Spence, 2016). Upon release from foods or beverages,

volatile compounds stimulate neurons in the olfactory mucosa via receptors in the nasal cavity

(Delime et al, 2016). Since flavor is the combination of aroma and taste, perception of volatiles

by orthonasal (sniff) and/or retronasal (mouth) pathways has been of particular interest in flavor

research (Puttanniah & Halpern, 2001; Halpern, 2004; Small, Gerber, & Hummel, 2005;

Bojanowski & Hummel, 2012). The orthonasal route of odor presentation involves odorants

travelling from the external environment, through the anterior nares, and towards the receptor

cells in the olfactory mucosa (Sun & Halpern, 2005; Roudnitzky et al., 2011). Conversely, the

retronasal route involves aromatic compounds from chewing food within the oral cavity

ascending through the oropharyngeal pathway to the olfactory mucosa. Retronasal olfaction is

usually accompanied by eating, drinking, swallowing and exhalation (Sun & Halpern, 2005).

Strong evidence has shown that odors are perceived differently when presented in ortho-

versus retronasal routes (Rozin, 1982; Buettner et al., 2008; Hummel & Heilmann, 2008).

However, retronasal olfaction has been studied less extensively mainly due to difficulties

encountered with regard to stimulus control (Heilmann & Hummel, 2004; Altundag et al., 2014).

Techniques for precise application of chemosensory stimuli via the retronasal route are limited.

Heilmann et al. (2004) developed a novel stimulation technique for controlled retronasal odor

application by releasing odors directly into the epipharynx, above the soft palate, which provided

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retronasal activation without concomitant gustatory activation. This method has now been widely

accepted for studying retronasal olfaction (Frasnelli, Heilmann, & Hummel, 2004).

The differences between ortho- and retronasal perception have been explained by many

theories, two of which are of particular interest to this review. The first of these is Max Mozell's

"chromatographic" model of olfaction. Mozell (1970) suggested that the process of olfactory

discrimination at the mucosal level is analogous to the process of chromatography, which is

commonly used in laboratories to separate and identify chemical compounds. He observed that

"…the molecules of one substance, when allowed to do so, will migrate along a liquid or solid

surface more rapidly and reach a given point in greater numbers per unit time than will the

molecules of another substance" (Mozell, 1970). This phenomenon has implications for

differences in odorant concentrations and airflow patterns. Rozin (1982) put forth the second

theory, which suggested that ortho- and retronasal olfactory perceptions are two different

systems. He argued that "olfaction is the only dual sensory modality, in that it senses both

objects in the external world and objects in the body (mouth)," and that "...the same olfactory

stimulation may be perceived and evaluated in two qualitatively different ways, depending on..."

its route of presentation. This experience has the capacity to greatly affect the perception of

flavor, and may therefore have implications within the food industry.

Numerous studies provide evidence that ortho- and retronasal odorant perceptions activate

different neural responses in diverse areas of the brain. For example, Small et al. (2004) reported

that when a taste is perceived simultaneously with a retronasally presented odor, the anterior

cingulate cortex (ACC), the orbitofrontal cortex (OFC), and the dorsal and ventral insula show a

superadditive response, and become more active. Conversely, when a taste is perceived

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simultaneously with an orthonasally presented odor, these same regions show significant

deactivation. Therefore, the ACC, OFC, and insula have all been implicated as key components

underlying flavor perception. With the use of functional magnetic resonance imaging (fMRI), it

has been proposed that both previous experience, as well as the route of olfactory delivery, have

an impact on taste-smell integration in the brain. It has also been suggested that different neural

recruitment occurs depending on whether the odorant represents a food or a non-food item

(Small et al., 2005). Taken together, these findings suggest that there are a number of factors

affecting brain activation with regard to the sense of smell.

Furthering our understanding of olfaction will be of great value to the food industry, and

benefit consumers alike. The aim of this article is to review recent findings and insights on

orthonasal and retronasal olfaction. The sense of smell has a profound influence on the

perception of flavor therefore an additional aim of this review is to shed light on the multifaceted

process of flavor perception, and the different roles that orthonasal and retronasal olfaction play

in this process. A variety of factors that affect the different routes of olfaction together with a

brief summary of the brain imaging techniques are discussed. This review summarizes recent

findings from the year 2000 onward, including important findings from the pioneers who set the

stage for this type of research.

2. Evidence of, and difference between flavor perception by ortho- and retronasal olfaction

Odors appear to be perceived differently when they are presented through the orthonasal

route as compared to the retronasal route. Olfactory stimulation resulting from the two modes of

olfaction is affected by different external influences and physiological factors, which is likely

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part of the reason why they have the potential to lead to diverse perceptions. For example, factors

like salivation, surface area, enzymes, chewing patterns, temperature changes, and especially the

textural properties of food items can affect flavor perception via retronasal pathways (Miettinen

et al., 2003; Diaz, 2004). These factors have the potential to affect flavor transport from saliva to

receptors, and alter volatile release patterns to the back of the throat (Taylor, 2002; Frasnelli, van

Ruth, Kriukova, and Hummel, 2005; Hummel et al., 2006; Buettner et al., 2008). Since these

factors should not impact orthonasal olfaction as it does not involve odors from within the body,

it may be considered more efficient with respect to flavor perception (Diaz, 2004). Pierce and

Halpern (1996) suggested that the variation in flavor perceptions via ortho- versus retronasal

routes may be the result of a difference in the efficiency with which odorants are delivered to the

olfactory mucosa.

One of the main setbacks in studying the differences between ortho- and retronasal olfaction

has been the lack of a controlled administration technique of odorous stimuli into retronasal

pathways. Since retronasal olfaction is closely related to taste, which involves other sensations in

addition to odor perception, flavor perception via the mouth is a more complex process and thus

harder to assess (Espinosa Diaz, 2004). As previously mentioned, Heilmann and Hummel (2004)

proposed a novel stimulation technique, in an attempt to allow for more precise control over

retronasal odorant stimulation, without the influences of simultaneous gustatory stimulation. In

this technique, odors are released directly into the epipharynx above the soft palate, thereby

eliminating the potential influences of chewing and swallowing. This method allows for

retronasal olfaction to be studied with an equivalent degree of control as that of orthonasal

olfaction, and ensures that measurements from the two locations may be evaluated in an

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unbiased manner.

2.1 Dose response behaviors, intensity ratings, and thresholds of odors

Different concentrations of a given flavor compound may be needed to evoke the same

aroma intensity, depending on whether the flavor is perceived by ortho- or retronasal olfaction

(Espinosa Diaz, 2004). The overall trend appears to be that orthonasal stimuli are generally

perceived as more intense than retronasal stimuli, even in blind subjects (Gagnon et al., 2003;

Heilmann and Hummel, 2004; Frasnelli, Heilmann, and Hummel, 2004; Sun and Halpern, 2005;

Hummel et al., 2006; Welge-Lussen, Husner, Wolfesnberger, and Hummel, 2009; Welge-Lussen,

Ebnother, Wolfensberger, and Hummel, 2009). This suggests that orthonasal thresholds to

odorants are lower than those of retronasal thresholds. Therefore, subjects require higher

retronasal odorant concentrations to detect an odor. It may be inferred then, that the retronasal

mode of olfaction is less effective for the perception and subsequent identification of odors as

compared to the orthonasal mode of olfaction. This effect was demonstrated by Sun and Halpern

(2005), when they found that odorants presented in the orthonasal location were almost always

correctly identified, whereas odorants presented in the retronasal location were accurately

identified less often.

In accordance with the ideas set forth by Mozell et al. (1970), it has been proposed that

retronasal presentation of an odor may produce a different pattern of mucosal activation than that

of orthonasal presentation of the same odor (Heilmann and Hummel, 2004; Sun and Halpern,

2005; Hummel et al., 2006). This could account for the more efficient perception of orthonasally

presented odors, and may help explain the discrepancies seen in odor thresholds. Other possible

explanations for this phenomenon need to be presented and tested, as it is likely that a

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combination of factors rather than one factor alone may be present.

2.2 Olfactory event-related potentials (ERPs) to ortho- and retronasal stimulation

Event-related potentials (ERPs) are a common electrophysiological measure used in the

study of orthonasal and retronasal olfaction. ERPs record the brain’s response to sensory

stimulation, and in this case by odorants. These recordings may be used to compare the response

amplitude, or strength of response, as well as the response latency, or the time taken to elicit a

response, in graphical form. Different olfactory ERP responses should be expected based on odor

stimulation site. Many investigators using this measurement and its data have found that smaller

amplitudes and prolonged latencies occur with respect to retronasal stimulation as compared to

orthonasal stimulation (Heilmann and Hummel, 2004; Frasnelli et al., 2004; Hummel et al.,

2006; Welge-Lussen et al., 2009; Roudnitzsky et al., 2011). This discovery relates back to the

finding that retronasal stimulation results in a weakened perception of odor intensity, along with

larger threshold values. While ERPs offer the advantage of not requiring full patient

collaboration, they are not routinely used as a diagnostic tool for the evaluation of olfactory

function, mainly because this technical procedure requires a stimulator capable of delivering a

brief, synchronized, and selectively chemosensory stimulus, which can be a challenge (Rombaux

et al. 2007).

As such, there are certainly situations in which the patterns of olfactory ERPs stray from

their usual norms. Hummel and Heilmann (2008) suggested that olfactory ERPs may differ

depending on the context in which the odor is presented. Specifically, they observed that a food

related odor (chocolate) produced a larger amplitude when presented orthonasally, but that a non-

food related odor (lavender) produced a larger amplitude when presented retronasally. This

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suggests that when odors are presented in “unusual site[s],” meaning sites in which they are not

typically perceived, the anticipated ERP patterns may be altered. Frasnelli et al. (2004) also

found that women show shorter response latencies than men, and commonly rate stimuli as being

more intense. It may be intriguing to explore the possibility of a gender difference, as it has the

potential to bring another aspect into the study of olfaction that has not received much focus thus

far.

ERPs have provided evidence to indicate that concomitant gustatory stimulation alters

odor perception via the two modes of olfaction in different ways. Welge-Lussen et al. (2009)

investigated the effect of simultaneous congruent and incongruent gustatory stimuli on olfactory

responses to two odors: vanillin and phenylethylalcohol (PEA). With orthonasal stimulation,

latencies were significantly shorter in the incongruent (sour-taste) condition, while during

retronasal stimulation, shorter latencies were produced in the congruent (sweet-taste) condition.

Interestingly, the simultaneous sweet gustatory stimulation influenced the food related odor,

vanillin, to a much more significant degree than that of the non-food related odor, PEA. With

respect to retronasal stimulation then, it is possible that the congruent stimuli were processed

more rapidly due to effects of familiarity. Conversely, it is possible that during orthonasal

perception, incongruent stimuli attract attention more quickly, leading to increased arousal and a

faster response time. Thus, the incongruent taste-odor pair might offer an explanation for the

protective role of the sense of smell, as it allows for the identification of potentially dangerous or

spoiled food. It would be valuable to perform additional research on the congruency of taste-odor

pairs, with a variety of novel odorants and tastants, to see if this effect can be generalized to

other situations.

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3. Dynamics of ortho- and retronasal aroma perceptions

3.1. Temporal and spatial dynamics of ortho- and retronasal aroma perceptions

When considering the dynamics of human perception of aromas, the perceived flavor

intensity as a function of duration and intranasal locations must be taken into account (Lee and

Halpern, 2013). Time - intensity tracking has been used to investigate taste and smell for many

years (Overbosch et al., 1986; Kuo, Pangborn, & Noble, 1993; Buettner et al., 2008; Blancher,

2012). With the improvement of stimuli control in the retronasal olfaction (Heilmann &

Hummel, 2004), the analytical concept of comparing temporal and spatial dynamics of ortho-

and retronasal olfaction was introduced (Buettner et al., 2008).

Lee and Halpern (2013) suggested the use of continuous stimulation and natural breathing

instead of presenting stimuli during a limited time frame using timed sniffs for time - intensity

tracking. They noted that ortho- and retronasally smelled intensity increased to their respective

maximum values at 11.7 and 8.1 s followed by gradual decreases to about half of the maximum

intensities within a 55 s tracking period. A similar early rise has also been reported for vanillin

(Stephenson & Halpern, 2009), citral (Kuo, Pangborn, & Noble, 1993), chocolate and lavender

(Heilmann & Hummel, 2004) odorants using ortho- and retronasal olfaction. More importantly,

retronasally-perceived initial, maximum and final intensities were found to be about half of that

experienced based on orthonasal smell, indicating potential adsorption or loss of flavor in the

retronasal pathway before reaching the olfactory mucosa. However, two airways did not show a

significant difference with regards to the initial time (1.4 ± 0.8 s) at which a nonzero intensity

was firstly indicated by the participant. Their data were comparable to others where a general 1 -

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2 s initial time has been previously reported (Laing & Macleod, 1992; Wise et al., 2003;

Soussignan et al., 2011; Møller et al., 2012).

Differences in the topographical distribution and adsorption of odorants by the nasal, oral

and pharyngeal mucosa, and the olfactory epithelium may affect the perceptual differences

between ortho- and retronasal olfaction. Frasnelli et al. (2005) measured intranasal odor

distribution in relation to four different nasal positons during oral administration of flavors using

an online-investigation tool proton-transfer reaction mass spectrometry (PTR-MS). A clear

intranasal gradient pattern was developed for the maximal amplitudes, which decreased in the

following order: nasopharynx > at the nostril > in front of the middle turbinate > olfactory cleft

(Frasnelli et al., 2005). Furthermore, spatial variations in concentration of different odorants

were observed indicating a significant interaction between “position” and “compound”. Buettner

et al. (2008) monitored the spatial and temporal distribution of the stimulus in vivo at defined

intranasal location using on-line PTR-MS analysis. They observed that the orthonasal-ipsilateral

response exhibited a sharper increase followed by an immediate drop, while a flatter profile was

noted for the orthonasal-contralateral response. The authors considered that the initial sharp peak

was due to the fact that stimulus compounds had not yet been absorbed by the nasal mucosa. In

contrast, the decrease of stimulus intensity measured contra-laterally during orthonasal olfaction

can be attributed to the fact that the stimulus spread and interacted along with the mucosal lining.

A similar trend was observed for retronasal olfaction via ipsilateral and contralateral

stimulations, but the difference noted was not as significant as that seen for the orthonasal

pathway. These observations further sustain Mozell’s theory in which the odorant was spread

along the olfactory mucosa in a chromatographic pattern (Mozell, 1970).

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3.2. Effect of dynamic physiological factors (processes) on ortho- and retronasal aroma

perception

In addition to the temporal and spatial differences during aroma perception, dynamic

physiological processes such as chewing, swallowing, breathing as well as air flow patterns play

an important role in elucidating perception of odors via orthonasal and retronasal routes

(Buettner et al., 2008). These physiological factors have been evaluated either as individual or

integrated processes.

Differing from orthonasal flavor perception, retronasal olfaction is strongly connected

with flavor or food intake, which is usually followed by swallowing (Welge-Lussen et al., 2009).

In real-time visualization of masticatory and swallowing processes, Buettner et al. (2008)

monitored oropharyngeal performance on retronasal ethyl acetate release during and after wine

consumption using real-time video fluoroscopy and PTR-MS. It is clear that during intake (a

small sip of wine), at which the velum was opened, a pulse of ethyl acetate was detected in the

expired air from the nose. However, no ethyl acetate was detectable in the breath when the sip

was taken but lips were closed. This effect was immediately removed when the taster opened

their velum under instruction inferring the critical role of airflow and breath during flavor

transfer and perception. A similar flavor release profile was noted during mastication of a solid

gel in which multiple peaks were visualized during chewing due to the intermittent opening of

the velum-tongue border (Buettner et al., 2001). Along with swallowing, a drastic increase of the

pulse of ethyl acetate was monitored for both wine and gel samples in a process has been

recognized as “swallow breath” (Buettner et al., 2001).

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The way participants chew samples also affected odor release patterns. Butettner et al.

(2008) found that a faster chewer had a quicker and steeper initial flavor release compared with

non-chewers, while non-chewers possessed a much higher “swallow breath” peak due to the

release of accumulated volatiles within the oral cavity prior to swallowing.

To fully elucidate the dynamics of ortho- and retronasal flavor perception, intranasal

airflow patterns on the perceptual differences of the ortho- and retronasal olfaction must not be

overlooked (Hummel et al., 2006; Buettner et al., 2008). Previous studies and development of

mathematical models have been conducted to mimic in vivo physiological conditions (Keyhani,

Schere, & Mozell, 1997, Zhao et al., 2004; 2006); however, the influence of air-flow

characteristics on olfactory perception has not been fully understood.

Breathing has been considered to be necessary for the aroma transfer during flavor

perception (Diaz, 2004; Buettner et al., 2008). The aroma that was detected at the nostrils during

an exhalation breath has been used to represent aroma at the receptor site (Taylor, 1996). Deibler

et al. (2001) proposed that food consumption was not a static equilibrium or a truly dynamic

system. Air flow rate is able to drive volatiles flowing back to the food matrix leading to an open

system during consumption (Deibler et al., 2001). It has been noted that less effective air

movement during expiration than inspiration could be one of the factors that lead to the

diminished response and the longer latencies for odorants presented retronasally (Scott et al.,

2007; Zhao et al., 2006).

Zhao et al. (2004) established a numerical model to anticipate the air flow and odorant

transport using commercial computational fluid dynamics (CFD) simulation techniques. They

suggested that small anatomical changes in the nasal cavity could strongly influence the airflow

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patterns and odor transportation which consequently affected the olfactory function (Zhao et al.,

2004). As a result, the potential anatomical variations during inhalation and exhalation

accompanied by the ortho- and retronasal olfaction could potentially account for the difference

observed between these two olfactory modes. In a continuous study, they simulated turbulent and

laminar airflow and odorant transport during human sniffing and found that sniffing behavior by

humans created a turbulent airflow, similar to laminar airflow during resting breathing (Zhao et

al., 2006).

3.3. Odor identification and localization

Identification and localization of olfactory stimuli have been utilized as effective tools to

provide further insight into understanding of the differences between ortho- and retronasal flavor

perception. Generally, identification of odorants appear to be much better when flavors were

introduced via orthonasal route compared to retronasal presentation (Pierce and Halpern, 1996;

Halpern, 2004), although some have found no significant difference (Sun and Halpern, 2005).

Using identification as a tool, Sun and Halpern (2005) systematically evaluated identification of

paired retronasal and orthonasal odorants. They noted that the perceived order of odorants were

the opposite of the physically presented sequence for both retronasal-first and orthonasal-first

heterogeneous pair stimulations. It was suggested that a short-term odorant memory could cause

odorant smelled second to be perceived before the odorant smelled first (Kellogg, 2003). Pfaar et

al. (2006) produced a mechanical obstruction of the anterior olfactory cleft of healthy subjects to

simulate the condition of nasal polyposis patients who were known to have superior retronasal

olfactory function (Landis et al., 2003). It was found that, after the olfactory cleft was blocked,

lower identification ability was noted for orthonasal olfaction but not for retronasal olfaction,

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which further indicates there is an absolute difference between orthonasal and retronasal

olfactory functions.

Localization of olfactory stimuli has been utilized to explore the qualitative differences

between ortho- and retronasally presented olfactory stimuli (Hummel et al., 2006). Hummel et al.

(2006) found that participants were able to successfully localize the trigeminal stimulant CO2,

pure olfactory stimulant H2S and phenyl ethyl alcohol. As the localization was found to be

independent of the stimuli intensity, the author considered that the localization was based on a

qualitative difference in the perception of ortho- or retronasal stimuli.

4. Impact of selected flavors and media on ortho- and retronasal flavor perception

4.1. Effect of flavor with different physicochemical properties

Critical emphasis has been placed on evaluating the association of physicochemical

properties of different odorants on ortho- and retronasal flavor perception (Diaz, 2004; Frasnelli

et al., 2005; Scott et al., 2007; Wikes et al., 2009), and some conflicting results have been

reported. Table 1 summarizes the recently published papers on the impact of selected flavor and

media on ortho- and retronasal flavor perception.

Diaz (2004) demonstrated a good correlation between water-to-air partition coefficients (Log

Pwa) of a homologous series of esters and the difference in ortho- and retronasal flavor

perception as indicated by retronasal: orthonasal ratio (R:O ratio). This ratio reflects the number

of times that flavor concentration needs to be elevated to obtain the same intensity by mouth as

by nose. It was found that hydrophobic flavor (lower air-water partition coefficient) produced a

higher R:O ratio and a larger difference in dose-response curve for ortho- and retronasal

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olfaction. It appears that a greater adsorption of highly hydrophobic flavors in the nasophrynx

may occur in the course of retronasal olfaction, which makes retronasal perception more

challenging.

Similarly, Frasnelli et al. (2005) measured the distribution of three different flavors at four

intranasal positions. To relate the flavor distribution pattern with the hydrophobicity of flavors,

they noted that the concentration of flavors at nasopharynx correlated with the hydrophobicity of

flavors (hydrophilic diacetyl (25%) < intermediate ethyl butyrate (31%) < highly hydrophobic

ethyl hexanoate (40%)) inferring flavor intranasal distribution may be dependent upon the

physicochemical properties of flavors. More importantly, not only spatial distribution of different

odorants varied at different intranasal sites, the respective flavor latency of maximal response

exhibited differently. Ethyl butyrate reached its maximal response at an average of 18.0 s and

then decreased; however, the response of diacetyl and ethyl hexanoate continued to increase and

reached their peaks 2.5 s and 5 s after ethyl butyrate, respectively. These observations suggest

that the picture evoked in the brain could be dynamically changed as each perceived flavor

concentration changed over time (Frasnelli et al., 2005). Considering the chromatographic theory

of Mozell (1970), the differences in latencies of maximal response further support that different

odorants might spread in different spatial and temporal patterns along the mucosa as a result of

their varying diffusion rates and solubility in mucus.

From another perceptive, Scott et al. (2007) observed that nonpolar or hydrophobic odorants

were more effective in activating rat olfactory receptors than polar or hydrophilic odorants in

retronasal stimulation; this effect was not dependent upon odorant concentration and carrier air

humidity (Scott et al., 2007). A potential mechanism which involves the removal of polar

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odorants from the air stream before entering olfactory receptor cells may lead to the differential

effect. In addition, a reduced transfer of polar odorants to the olfactory region due to less

effective air movement during expiration than inspiration could be another factor (Zhao et al.,

2006; Scott et al., 2007).

Instead of analyzing the polarity (water-air partition coefficient) of odorants themselves,

Wilkes et al. (2009) suggested that it was the solubility of odorants in mucus rather than in water

(polarity), which contributes to the perceptual difference of ortho- and retronasal olfaction. In

their analysis, paired odors with known solubility in nasal mucus and water were presented via

both ortho- and retronasal routes and the odorant that was identified first in the binary mixture

was recorded. A temporal difference was observed for retronasal olfaction whereby the odorants

with lower mucus solubility were significantly easier to be identified first and recognized

correctly. This finding strongly suggested that, in retronasal olfaction, the odorants with higher

mucosa solubility may undergo a greater adsorption effect at the nasophrynx such that they are

perceived more slowly than the odorants possessing lower mucus solubility at the nasal mucosa.

In other words, the response evoked at olfactory receptors is a function of the degree of

absorption and desorption rate of odorants along the way through respiratory tract and the

resulting number of flavor molecules that reached the olfactory respecters. These findings sustain

the Rozin (1982) theory of flavor perception which proposed that selective adsorption and

desorption of odorants by the mucosa in mouth and nasopharynx in comparison to the nose

during orthonasal perception may account for the qualitative difference and duality in the ortho-

and retronasal olfactory senses.

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4.2. Effect of trigeminal stimuli

In addition to activation of olfactory nerves, it is known that most odorants can stimulate

the activation of the trigeminal system; an effect that is mainly concentration dependent (Doty et

al., 1978). The entire nasal mucosa can be involved in the intranasal trigeminal stimulation

(Stevens & Cain, 1986), while olfactory receptors are mainly located in the area of olfactory cleft

(Leopold et al., 2000).

Hummel et al. (2006) compared the olfactory ERPs of a trigeminal chemosensory stimulant

(CO2) and intranasal mechanical stimuli (air puffs) at different intranasal locations. It was found

that the chemosensory stimulus was more strongly perceived and had short latency when

stimulus was introduced orthonasally compared to retronasally, while a reverse pattern was

observed for the mechanical stimuli. Similar patterns were reported by Frasnelli, Heilmann and

Hummel (2004) who concluded that the responsiveness of human nasal mucosa to trigeminal

stimuli are dependent upon the quality of stimulus (chemosomatosensory vs. mechanosensory

stimuli) and the site of stimulation (orthonasal vs. retronasal). They outlined that the respiratory

mucosa should be seen as a heterogeneous tissue whose differences may contribute to the

dissimilarity during orthonasal and retronasal odorous stimulation (Hummel et al., 2004; 2006).

Frasnelli, Ungermann, and Hummel (2008) further investigated the role of trigeminality of

stimulus in localization (orthonasal vs. retronasal) and lateralization (left vs. right) of orthonasal

and retronasal stimuli. They revealed that the degree to which an odorant activates the trigeminal

nerve plays a significant role in localizing and lateralizing intranasal stimuli and is more

pronounced for lateralization (Frasnelli, Ungermann, and Hummel, 2008). However, no direct

correlation was found between subjects’ ability to lateralize and localize stimuli. For the odors

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with minimum trigeminal properties, localization was more accurate than lateralization. This

further infers that trigeminal stimulation might be more important for lateralization.

Additionally, based on Mozell’s chromatographic model of olfaction (Mozell 1970), different

zones of olfactory epithelium would be activated in a reverse order when odorants enter nasal

cavity from the orthonasal or retronasal routes. This difference could also contribute to the flavor

localization effect (Frasnelli et al., 2005; Scott et al., 2007).

4.3. Effect of food texture

Water, protein, and fat in food systems interact with one another and can influence volatile

release patterns. The course of ortho- and retronasal odor perception during and after food

consumption is not only influenced by the odorants but also physiochemical properties such as

viscosity and texture of food matrices. As the release of flavor molecules from the food matrix

into the headspace (air or in mouth) directly relates to the amount of flavor molecules that are

able to reach the olfactory receptor during eating, the factors that would affect the phase

partitioning and mass transportation of flavors need to be taken into account in the context of

dynamic flavor perception. Table 1 illustrates several recent studies in which emphasis has been

placed on understanding the interactions between texture and olfactory stimuli using

psychophysical and electrophysiological approaches.

It has been previously reported that viscosity (Pangborn & Szczesniak, 1974; Hollowood,

Linforth, & Taylor, 2002; Saint-Eva et al., 2006), texture (Weel et al., 2002; Bult, de Wijk, &

Hummel, 2007), oral shear stress (Cook et al., 2003), surface exchange area of food matrices

developed in mouth and throat (Saint-Eva et al., 2006) and addition of gelling agents (Juteau,

Tournier, & Guichard, 2004) affect flavor perception.

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Frasnelli et al. (2005) determined temporal profiles of volatile odorants at different

intranasal locations when consuming liquid and solid custards using PTR-MS. An interaction

was detected between the type of custard and compound, which inferred the release of different

compounds from the custard during oral administration was significantly affected by viscosity.

In addition, the total number of compounds measured for solid custard was 34% less than the

amount of flavors released from the liquid sample indicating lower viscosity results in higher

flavor release. Buettner et al. (2008) measured the impact of gel texture on ethyl butanoate

release during mastication and swallowing. A fast and intense onset of aroma transfer to the nose

was observed for a soft gel (4% protein content) in comparison with a hard gel (10% protein

content) due to a faster breakdown of soft gel structure resulting in a faster release of volatiles

(Buettner et al., 2008). However, in a recent study, Roudnitzky et al. (2011) observed that

thickened milk enhanced the intensity of a butter aroma, which was contradictory with previous

reports in literature (Hollowood, Linforth, & Taylor, 2002; Frasnelli et al., Saint-Eva et al., 2006;

Bult, de Wijk, & Hummel, 2007; Buettner et al., 2008). The authors attributed the difference to

the sweet taste of oral stimulus inducing a taste-aroma interaction and potential confusion by

panelists when rating odor intensity (Roudnitzky et al., 2011). A subsequent study revealed that

increased milk viscosity and simultaneous presentation of a creamy aroma ortho- and

retronasally enhanced odor intensities (Iannilli et al., 2014).

4.4 Effect of visual cues

Sight is undoubtedly critical for the enjoyment and perception of foods and the impact of

visual cues on olfactory perception has been well documented. Visual cues, like color, can

dominate over people’s olfactory experience of foodstuffs, and can vary greatly among

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individuals due to differing flavor expectations (Spence, 2016). When colors accompany odors,

perceptions of the odors change and appropriate colors often impact the perception of the odors

differently from inappropriate colors (Zellner et al, 2013). Koza et al. (2005) demonstrated that

color has a qualitatively different effect on the perception of the orthonasally compared to

retronasally presented odors. They found that when red dye was added to a fruit-flavored

beverage, the intensity of the drink’s aroma was rated higher in those who assessed the sample

orthonasally. Another group of participants rated the drink as less intense in the presence of red

color when they were experienced retronasally. This suggests that assumptions about the

association between olfactory perception and visual cues may not necessarily be the same when

participants actually taste the product. When vision is taken out of the equation, for instance in

the case of congenitally blind subjects, olfactory perception is generally enhanced while taste is

diminished. Gagnon et al. (2015) confirmed that blind subjects have enhanced orthonasal but not

retronasal olfactory abilities compared to their sighted counterparts. It is likely that familiarity of

odorants, but also the absence of visual cues play a role in this observation.

5. Methods to understand ortho- and retronasal flavor perception by neural and brain

imaging techniques

Odorous stimuli reach the olfactory epithelium via one of two routes. The first is through

retronasal stimulation during food ingestion via the mouth and nasopharynx, and the second is by

sniffing in through the external nares of the nose during orthonasal stimulation. Paul Rozin

(1982) suggested that the sense of smell is the sole “dual sensory modality,” in that it senses

odors from within the body (retro), and from the external environment (ortho). There is a

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growing body of evidence to support the contention that these two modes of olfactory perception

are, in fact, separate systems.

There are different types of stimuli associated with the two routes of olfactory perception.

Retronasal stimuli are generally limited to food volatiles, which likely contributes to the reason

why retronasal olfaction is commonly mistaken for “taste.” On the contrary, orthonasal stimuli

consist of a wide variety of scents and smells, including food aromas, but also perfumes, floral

scents, and pheromones among others (Shepherd, 2006).

Many studies have shown that different paths of odorant delivery, along with experience,

may play a role in influencing which areas of the brain become active during olfactory

perception (Small et al., 2004; Small, Gerber, Mak, & Hummel, 2005; Bender, Small, Hummel,

& Negoias, 2009). In relation to taste-odor integration, Small et al. (2004) reported that when a

taste is perceived simultaneously with a retronasally presented odor, the anterior cingulate cortex

(ACC), the orbitofrontal cortex (OFC), and the dorsal and ventral insula show a super additive

response, and become more active. Conversely, when a taste is perceived simultaneously with an

orthonasally presented odor, these same regions show significant deactivation. These conflicting,

path-dependent responses seemed to be correlated with whether or not the taste-smell pair

seemed congruent based on prior learning.

Further evidence for differential neural processing of odors based on route of odorant

delivery is given by Bender et al. (2009). They found that the effects of salivary habituation,

which occurs after repeatedly presenting a single food odor through a single route, and results in

a decrease in salivary response, are reversed when the route of presentation of that odor is

switched. This demonstrates that a switch in odorant delivery from ortho- to retronasal, for

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example, allows for recovery from habituation, and implies diverse brain activation for each

route. In preliminary studies conducted by Hummel et al. (2006), fMRI data revealed neural

activation in the insula, anterior cingulate, superior temporal gyri, and right lateral orbital gyrus,

regardless of the site of stimulation. However, when the site of stimulus presentation was taken

into account, cortical activation in the left insula and anterior cingulate was higher following

orthonasal stimulation, thus providing possible evidence of site-specific brain activation patterns.

5.1 Utilization of brain imaging techniques in comparing responses to food and non-food odors

It has been speculated that previous experience with certain odors may affect their

processing (Small et al., 2004; Small et al., 2005). A more specific example of this phenomenon

is seen when the odor in question is associated with a food, or a non-food odor (Heilmann &

Hummel, 2004; Small et al., 2005; Hummel & Heilmann, 2008). Odors associated with foods are

normally experienced both ortho- and retronasally, while odors associated with non-foods tend

only to be experienced orthonasally. The differential processing seen in these situations may

therefore be attributed to cues associated with smelling a food item versus a non-food item. As

pointed out by Bender et al. (2009), orthonasal perception of a food odor signifies the availability

of food, while retronasal perception of the same odor acts as an indication that food has actually

been ingested. This connects to the concept of food reward, which may have the potential to

affect neural recruitment since retronasal perception of food odors implies the receipt of food (a

reward), whereas that of non-food odors does not. For instance, a recent study demonstrated that

the delivery of strawberry aroma compared to lily of the valley highlighted reward-related areas

in the brain (Iannilli et al, 2015).

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In the research conducted by Bender et al. (2009) discussed above, they also found that a

recovery from salivary habituation occurs when the odor is switched. However, this effect was

only observed when the odor in question was food related, thus providing further support for

differential representation of food versus non-food odors. Similarly, with the use of fMRI, Small

et al. (2005) found that activation of the perigenual cingulate, posterior cingulate, medial OFC,

and superior temporal gyrus occurred in response to a food odor (chocolate) via the retronasal

route. However, orthonasal perception of the same chocolate odor resulted in favorable

activation in very different regions, namely the insula, hippocampus, caudolateral OFC,

thalamus, amygdala, and the temporal, parietal, and frontal opercula. They also found that this

result was not replicated in response to non-food odors, such as lavender. This, again, suggests

that brain activation may vary with the route of odorant delivery, but that this effect depends on

whether the odor in question has been experienced by one or both modes of olfactory perception

in the past. Gautam and Verhagen (2010) pointed out that previous experience with a taste-odor

pair may lead to specific taste acquisition by odorants. Orthonasal perception does not have a

taste component, so this taste acquisition phenomenon may help to explain why different brain

regions become active in response to food versus non-food odors. It is much more probable that

a food odor like chocolate has been previously experienced both ortho- and retronasally than has

a non-food odor like lavender, since it is less customary to ingest lavender than it is to ingest

chocolate.

6. Conclusions

Although much has been uncovered with the advancement of neural and brain imaging

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techniques, more research is needed to replicate the findings. With each new study, we gain a

better understanding of the complexities of ortho- and retronasal olfaction, and the underlying

network of brain activation during olfaction. Because olfaction has a significant influence on the

types of foods and beverages that appeal to consumers, expanding our knowledge on the subject,

along with clarifying the presence of gender differences, has the potential to benefit the food

industry through the development of more successful and pleasant products on the market. It

would also be beneficial to perform similar experimental tests with an array of different odors to

see if the effect seen in the studies discussed in this review applies in more than one situation.

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Table 1. Summary of recent studies of impact of selected flavors and media on ortho- and retronasal flavor

perception

Authors/years, Research objectives Design of Methodology Conclusions

experiment

Effect of physicochemical properties of flavors on orthonasal and retronasal perception of flavors

Diaz, M. (2004) To compare 15 subjects Dose Reponses and Miglyol- Orthonasal and retronasal

orthonasal vs. evaluated intensity air partition coefficients of perception of flavor depends

retronasal dose- using a scaling flavor were monitored using on physical characteristics of

response behavior of methodology. supra-threshold-olfactometer flavors.

a homologous series Perceived intensity and headspace GC.

of esters of flavors was

plotted against

concentration.

Measured intranasal

odor concentrations Used proton-transfer

To determine the
at least twice in 10 reaction-mass spectrometry
temporal profiles of Different volatile flavor
Frasnelli et al. participants at 4 to determine concentrations
volatile odor compounds exhibit different
(2005) nasal positions at the four different location,
concentrations at temporal and spatial profiles
during the as well as latency of the
different locations in in their intranasal distribution.
consumption of swallow (for 9 participants
the nasal cavity.
liquid and solid out of 10).

custards.

Scott et al. To predict if odorant Rats were presented Odor stimulation to rats’ Nonpolar odorants were

(2007) polarity would be a to a series of external nares by artificial effective in both stimulus

major factor in odorants that varied sniff and to internal nares by modes, while polar odorant

orthonasal vs. from very polar, positive pressure. activated olfactory epithelium

retronasal olfaction. hydrophilic to very Electroolfactogram was used presented orthonasally.

nonpolar, to record the responses.

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hydrophobic

odorants.

Wilkes et al. To determine 33 subjects were Psychophysical method: Solubility of odorant in mucus

(2009) whether solubility of exposed to 4 pairs either sniffing or swallowing rather than in water better

odorant in mucus or of odorants that air above sample into mouth predicts which odor will be

water predicts represent same or and identify which odor was perceived first.

temporal perception different solubilities perceived first and rate

of odors via in nasal mucus & intensity

retronasal route water

Delime et al. To evaluate relative 100 naïve subjects Odor Activity Values Compared to retronasal

(2016) contributions of evaluated the (OAVs) reported in the perception, orthonasal

volatiles in relative literature were compared to perception was more sensitive

orthonasal vs. contributions of d′ values established in this to the removal of all

retronasal olfaction volatiles in a nine- study to determine if OAVs individual volatiles, except for

to overall perceived component, were representative of the 2,3-butandione and methyl

strawberry flavor commercial individual contribution of dihydrojasmonate. OAVs did

strawberry flavor volatiles to the perceived not always reflect the relative

both ortho- and strawberry flavor. importance of individual

retronasally. volatiles in flavor represented

by Thurstonian measure ′.

Effect of physicochemical properties of medium on orthonasal and retroasal perception of flavors

Frasnelli et al. Measured intranasal Used proton-transfer An interaction of “compound”

To determine the
(2005) odor concentrations reaction-mass spectrometry and “type of gel” was noted.
temporal profiles of
at least twice in 10 to determine concentrations Liquid gel resulted in higher
volatile odor
subjects at 4 nasal at the four different loci, as flavor release compare with
concentrations at
positions during the well as latency of the solid gel.
different locations in
consumption of swallow (for 9 participants
the nasal cavity
liquid and solid out of 10).
during consumption
custards.

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of liquid and solid

custard samples.

Buettner et al. Intensity and Used proton-transfer Solid gel had a lower
To elucidate
(2008) temporal reaction mass spectrometry maximum sensory intensity
relationship between
characteristics 4 and to measure total amounts of compared with soft gel.
molecular level and
10% protein gel odor detected, and odor
perception during
were rated by 10 intensity.
and after
assessors.
administration of a

chemical stimulus.

Roudnitzky et To understand Odor and texture ERPs were obtained from Perceptual interactions were

al. (2011) interactions between intensity of lean and five recording positions found between food texture

synchronous tactile thickened milk were using psychophysical and and odor.

(texture) and rated by 18 subjects electrophysiological

olfactory (odor) either orthonasally approaches.

sensations or retronasally.

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Direct comparison of orthonasal and retronasal perception of flavors.

Heilmann To develop a new Food-related (chocolate) Used a computer-controlled air- There are significant

& method to compare and non-food (lavender) dilution olfactometer to allow differences between

Hummel. orthonasal and retronasal related flavor with application of rectangular- ortho- and retronasal

(2004) olfaction. controlled conc. were shaped pulses of chemical perception of odors,

applied in either anterior stimuli. on both threshold and

nasal cavity or suprathreshold levels.

epipharynx. Stimuli

concentration and

olfactory responses were

recorded.

Frasnelli, To compare responses 40 subjects underwent Chemosomatosensory ERPs Interactions were

Heilmann, obtained by either were recorded using gaseous found between

& electrophysiological and chemosomatosensory CO2 as a stimulant; stimulus properties

Hummel psychophysical methods stimulation using gaseous mechanosensory ERPs were and site of

(2004) in response to CO2 or mechanosensory recorded in response to stimulation.

mechanical and stimulation using air puffs. intranasal mechanical stimuli Chemosensory

chemosensory (air puffs). A computer stimuli were more

stimulation to anterior or controlled olfactometer was sensitive to

posterior of nasal cavity. used to control stimulus orthonasal anterior

presentation. position, while

mechanical stimuli

were more sensitive

to retronasal posterior

position.

Hummel To investigate 230 individuals were Used Sniffin’ Sticks test and There are perceptual

et al. differences in odor assessed for ortho- and grocery store test; then released differences in relation

(2006) quality between ortho- retronasal olfactory odors into anterior portion of to ortho- and

and retronasal olfaction functions. nasal cavity (for orthonasal) retronasal stimulus

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in isolation, using and into epipharynx of soft presentation (related

psychophysical, palate (for retronasal) for more to air flow patterns,

electrophysiological, and defined stimulation (under contextual effects...).

imaging techniques. endoscopic control).

Hummel To investigate the 20 healthy subjects were Tubing was placed below the Food related odor was

& perception of odor exposed to the non-food lower turbinate (for retronasal more actively

Heilmann. intensity following (lavender) and food stimulation) and in the perceived in the

(2008) ortho- and retronasal (chocolate) flavor, while vestibulum of the nasal cavity orthonasal route. The

odor presentation and to measuring ERPs. (for orthonasal stimulation). opposite occurred for

determine olfactory ERP was measured non-food related odor.

ERPs in response to electrophysiologically. Flavor perception is

ortho- and retronasal dependent on context

stimulation with odors and route of odor

related or unrelated to presentation.

foods.

Role of brain imaging and signal to characterize orthonasal and retronasal flavor perception

Rozin To prove that olfaction is N/A Discussed various past There is difficulty in

(1982) the only dual sensory experiments to explain duality identifying the flavor

modality. of senses. of an identified

odorant, thus

suggesting duality in

olfaction.

Small et To investigate the role of 20 participants were 11 participants (3 men, 8 The insula, OFC, and

al. (2004) experience in forming presented with 2 flavors – women with mean age of 26) ACC are key

the neural representation one containing a progressed to neuroimaging components of the

of flavor (taste-odor congruent taste-odor pair stage. Used event-related fMRI network underlying

pairings), and whether (vanilla/sweet), and the to evaluate brain response flavor perception and

the mode of olfactory other an incongruent pair during perception of flavors taste–smell

delivery affects (vanilla/salty), and a compared with the sum of the integration within

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taste/smell integration. tasteless-odorless independent presentation of these and other

baseline. their constituents (via regions. It is

retronasal route). dependent on mode of

olfactory delivery and

previous experience

with taste/ smell

combinations.

Small et To evaluate differential A two factorial design Delivered vaporized odorants Routes of delivery

al. (2005) neural responses evoked with odor (lavender, via the orthonasal and produced differential

by orthonasal versus butanol, farnesol and retronasal routes and measured activation in brain

retronasal odorant chocolate) and mode of brain response with fMRI. tissues, inferring

delivery and to delivery (ortho and Neuroimaging was taken at interaction of route

determine whether this retronasal). This resulted each condition and pre- and and odorant. This

differential activation in eight odor conditions, post-processed. effect was influenced

varies with each with its own odorless by whether an

physicochemical baseline condition. odorant represented a

properties of odor. food or non-food

odor.

Shepherd, To investigate advances N/A Use of high-resolution fMRI. Human brain’s flavor

G. M. in brain mechanisms of system is closely

(2006) smell perception and the connected with eating

flavor system in the behavior and can

human brain evaluate and regulate

food intake.

Bender et To test the prediction that 24 subjects were exposed Evaluated pleasantness and Salivary response to

al. (2009) recovery from salivary to chocolate and pineapple intensity, familiarity and food odors decreases

habituation occurs when odors. perceived edibility. Used nasal with repeated

subjects are presented cannulas and olfactometers to presentation; this

with a novel food odor deliver odors. response rebounds

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(“odor-switching”) and upon presentation of a

when presented with the novel food odor via

same food odor via a the same route and

novel route (“route- upon presentation of

switching”). the same food odor

via a novel route.

Iannilli et To study the Two concentrations of each Food-related odors

18 subjects assessed
al. (2015) electrophysiological aroma were delivered via a (strawberry)
strawberry and lily of the
response to food and computer-controlled compared to an odor
valley aromas.
non-food related odors. olfactometer based on the with similar

principles of air-dilution. The characteristics but not

lower concentrations were used associated with food

for odorous stimulation during (lily of the valley)

electroencephalogram (EEG) highlights reward-

recordings. related brain

networks.

Interactions between olfaction and other factors related with ortho- and retronasal perception

Mozell To determine whether 24 frogs “sniffed” 16 The relative retention time of The olfactory mucosa

(1970) different mucosal different odorants, each at each of the 16 odorants was behaves like a polar

gradients of activity are four concentrations and measured in a gas chromatographic

established for different two flow rates. chromatograph. column.

odorants.

Sun & To study the 20 subjects were tested for Odorant intensity matching and Processing of

Halpern identification of paired identification of retronasal identification learning was retronasal and

(2005) retronasal and orthonasal and orthonasal only learned and trained to subjects. orthonasal odorants

odorants. odorants, homogeneously differ, and odor

and heterogeneously memory may play an

paired odorants. important role in

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flavor perception.

To determine the impact 15 subjects rated the Subjects were familiarized with Presence of color
Koza et al.
of color on orthonasal intensity of the odor of samples using practice enhanced odor
(2005)
and retronasal odor tangerine-pineapple-guava solutions first, followed by test intensity ratings for

intensities beverage with or without solutions and odorant intensity orthonasal olfaction,

red food coloring was judged according to the and decreased

intensity of the fruitiness using intensity ratings for

a 101-point scale. retronasal olfaction.

Welge- To examine whether 32 subjects took part in 2 Used olfactory ERPs as a Ortho- and retronasal

Lussen et congruent and test sessions. Four measure in response to a food- olfactory stimuli are

al. (2009) incongruent randomized blocks of 15 like odor (vanillin) and an odor processed differently;

simultaneous gustatory stimuli each were applied not normally associated with congruent and

stimuli influence either retro- or food (rose-like incongruent

orthonasal and retronasal orthonasally, while phenylethylalcohol). concomitant gustatory

odorant perception. simultaneously applying stimuli appear to have

sweet or salty gustatory a different impact on

stimuli. orthonasal and

retronasal olfaction.

Welge- To examine if retronasal 50 healthy subjects were Olfactory stimulation was Swallowing took

Lussen et olfaction influences presented with a sweet conducted using a computer- place significantly

al. (2009) swallowing in a manner taste (glucose) controlled olfacometer. faster and more

different from that of simultaneously with Gustatory stimulation was frequently after

orthonasal olfaction in vanillin, either ortho- or performed by a taste dispenser. retronasal stimulation

combination with retronasally. The intensity Ultrasound image of the mouth compared with

simultaneous gustatory rating of ortho- and floor was videotaped to swallowing after

stimuli. retronsal stimuli and continuously monitor orthonasal

frequency of swallowing swallowing activity. stimulation.

was recorded.

Gautam To test odor-taste Flavor-naïve rats were The retronasal odor stimulation Only after prior

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and generalizations in rats placed into test chambers was conducted by licking the paired experience of

Verhagen with or without paired to generalize of taste/odor odorized water from the spout the odorant with a

(2010) exposure to an odorant aversion. 2 oral odorant by rats sweet tastant, rats

and a tastant. solutions, benzaldehyde behaved towards

and amyl acetate together tasteless retronasal

with 4 tastants: sucrose, odorants as if they

NaCl, HCl and quinine were sweet.

HCl were used. The

number of licks by rats

was recorded.

Gagnon et To test the hypothesis 12 congenitally blind and For orthonasal olfaction, Blind subjects were

al. (2015) that congenitally blind 14 sighted control subjects subjects had to select one of significantly faster

subjects have enhanced were asked to identify four possible choices after and tended to be

orthonasal but not odors using grocery- sniffing samples blinded. For better at identifying

retronasal olfactory available food powders. retronasal identification, two odors presented

skills. mL of stimulus powder was orthonasally. Both

placed on the tongue using a groups performed

teaspoon, while the subject had better for retronasally

his/her nostrils occluded. After presented odors, but

stimulus delivery, the this gain was less

participant was asked to close pronounced for blind

his/her mouth, unblock his/her subjects.

nostrils, breathe normally and

identify the odorant while the

experimenter recorded response

time.

fMRI: high-resolution functional magnetic resonance imaging

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ERPs: Event-related potentials

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