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Flavor Perception
Edited by
ANDREW J. TAYLORProfessor of Flavour Technology
University of Nottingham, UK
DEBORAH D. ROBERTSFood and Flavor Science Consulting LLC
Chesterfield, Missouri, USA
Flavor Perception
Flavor Perception
Edited by
ANDREW J. TAYLORProfessor of Flavour Technology
University of Nottingham, UK
DEBORAH D. ROBERTSFood and Flavor Science Consulting LLC
Chesterfield, Missouri, USA
� 2004 by Blackwell Publishing Ltd
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First published 2004 by Blackwell Publishing Ltd
Library of Congress Cataloging-in-Publication Data
Flavor perception/[edited by] Andrew J. Taylor and Deborah D. Roberts.
p. cm.
Includes bibliographical references and index.
ISBN 1-4051-1627-7 (printed case hardback : alk. paper)
1. Taste. 2. Flavor. 3. Chemical senses. 4. Chemoreceptors. I. Taylor, A. J. (Andrew John).
II. Roberts, Deborah D., QP456.F56 2004
152.1’67–dc222003026666
ISBN 1-4051-1627-7
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Contents
Contributors xii
Preface xv
1 Measuring proximal stimuli involved in flavour perception 1
ANDREW J. TAYLOR and JOANNE HORT
1.1 Factors influencing flavour perception 1
1.1.1 Perception of ‘taste’ and its location 1
1.1.2 Multimodal nature of flavour perception 2
1.1.3 Distal and proximal stimuli 4
1.2 Aroma 4
1.2.1 Measurement of orthonasal proximal stimuli 5
1.2.2 Measurement of retronasal proximal stimuli 6
1.2.3 Examples of differences in distal and proximal
aroma stimuli 10
1.2.4 Relating proximal aroma stimuli to flavour perception 11
1.3 Taste 14
1.4 Texture 18
1.5 Colour and appearance 22
1.5.1 Colour perception 23
1.5.2 Specifying colour 24
1.5.3 Instrumental measurement of colour 27
1.6 Methods to study and quantify crossmodal interactions 29
1.7 Conclusion 34
2 The role of oral processing in flavour perception 39
JON F. PRINZ and RENE DE WIJK
2.1 Introduction 39
2.2 Anatomy of the peri-oral structures 41
2.2.1 Saliva 44
2.3 Flavour 46
2.4 Oral processing 47
2.5 Conclusion 53
3 The cellular basis of flavour perception: taste and aroma 57
NANCY E. RAWSON and XIA LI
3.1 Introduction 57
3.2 Taste and flavour 57
3.2.1 Taste buds and taste cells 58
3.2.2 Molecular mechanisms 60
3.2.3 Salt taste 60
3.2.4 Sour taste 62
3.2.5 Sweet taste 63
3.2.6 Bitter taste 64
3.2.7 Umami taste 64
3.2.8 Individual variations – polymorphisms in receptors 66
3.3 Perception of aroma 67
3.3.1 What is the ‘signal’? 67
3.3.2 How is odorant information encoded? 69
3.3.3 Odour intensity and quality coding in the olfactory bulb 70
3.4 Flavour perception can be modulated: adaptation,
sensitisation and crosstalk 74
3.5 Flavour perception and ageing 76
3.5.1 Anatomy and physiology 76
3.5.2 Taste 77
3.5.3 Olfaction 78
3.6 Conclusion 79
4 Structural recognition between odorants, olfactory-binding
proteins and olfactory receptors – first events in odour coding 86
JEAN-CLAUDE PERNOLLET and LOIC BRIAND
4.1 Introduction 86
4.2 Anatomy of the olfactory system 89
4.3 Olfactory-binding proteins 90
4.3.1 OBP discovery 90
4.3.2 General properties of vertebrate OBPs 91
4.3.2.1 Structural properties of OBPs 91
4.3.2.2 Comparison with insect OBPs 95
4.3.3 Experimental approaches to OBP–odorant interactions 96
4.3.4 Human OBPs 97
4.3.5 Structure of the odorant-binding pocket 98
4.3.5.1 Crystallographic observations of complexes
of OBPs with odorants 99
4.3.5.2 Biochemical molecular modelling and
docking investigation of hOBP 99
vi CONTENTS
4.3.6 Potential role of OBPs in odour discrimination 100
4.3.7 Conclusions about OBP 102
4.3.7.1 Vertebrate OBP definition 102
4.3.7.2 Comparison of OBP numbers in vertebrates
and in insects and OBP role in odorant
discrimination 102
4.3.7.3 Biological role of OBPs 102
4.3.7.4 Putative roles of OBPs 103
4.3.7.5 Lipocalins potentially involved in taste 104
4.4 Olfactory receptors 104
4.4.1 Discovery of ORs as G protein–coupled receptors 104
4.4.1.1 General properties of GPCRs 105
4.4.1.2 GPCR classification 106
4.4.1.3 G proteins as transducers 108
4.4.2 Peculiar properties of ORs 110
4.4.2.1 The second external loop of ORs 110
4.4.3 Odorant recognition by ORs: structural relationships
between OR and odorant 112
4.4.3.1 Experimental studies of odorant–OR interactions 112
4.4.3.2 OR functional studies reveal broad odorant
selectivity 114
4.4.4 Odorant-binding site in the receptor transmembrane
bundle 118
4.4.5 Conclusions on the odorant–OR structural relationships
and odotope definition 120
4.4.6 OR classification and genome comparison 121
4.4.6.1 Classification of the OR gene family 121
4.4.6.2 OR nomenclature 122
4.4.6.3 Variability of ORs within the human species 123
4.4.6.4 Comparison of the OR number between man
and animal and between vertebrates and insects 123
4.4.6.5 Human OR genome compared to macrosmatic
animals 124
4.4.7 Other possible functions for ORs 125
4.4.8 Concluding remarks about ORs 126
4.5 Biochemical mechanisms involved in odorant capture 127
4.5.1 Interactions of OBP with OR 127
4.5.2 Signal transduction in olfactory neurons and neural
impulse formation 129
4.5.2.1 Receptor activation and downstream signalling 130
4.5.2.2 OR desensitisation 130
4.5.3 Beyond the olfactory neuron 131
CONTENTS vii
4.6 Conclusion 132
4.6.1 Complexity of stereochemical odorant recognition
and subsequent odour coding 132
4.6.2 Taste and VNO receptors compared to ORs 134
4.6.3 Comparison of human and animal olfactory systems 135
4.6.4 Inferences from the recent knowledge about olfaction
pericellular events and future progresses 136
4.6.4.1 Odotope mixture cannot be distinguished from
odorant mixture 136
4.6.4.2 Human individual variability 137
4.6.5 Olfactory biosensors 138
5 Oral chemesthesis: an integral component of flavour 151
BARRY G. GREEN
5.1 Overview 151
5.2 Introduction 151
5.3 The neurophysiological basis of oral chemesthesis 152
5.4 Psychophysical characteristics 155
5.4.1 Sensation quality 155
5.4.2 Spatial factors in sensitivity and sensation quality 156
5.5 The roles of sensitisation and desensitisation in chemesthetic
perception 158
5.6 Temperature and chemesthesis 161
5.7 Interactions with touch 162
5.8 Interactions with taste and smell 162
5.9 Individual differences 165
6 Flavour perception and the learning of food preferences 172
ANTHONY A. BLAKE
6.1 Introduction 172
6.2 Flavour as an example of molecular communication 173
6.2.1 The human brain 174
6.2.2 Multisensory perception 176
6.3 What flavour is and how we learn to like it 177
6.3.1 Learning to like flavour 183
6.3.2 Flavour learning in adults 191
6.4 Conclusion 197
viii CONTENTS
7 Functional magnetic resonance imaging of human olfaction 203
MARTIN WIESMANN, BIRGIT KETTENMANN and
GERD KOBAL
7.1 Introduction 203
7.2 The methodological basis of functional magnetic resonance
imaging 203
7.2.1 Magnetic resonance imaging 203
7.2.2 Functional magnetic resonance imaging 204
7.2.2.1 Exogenous contract agent injection 205
7.2.2.2 Arterial spin labelling 205
7.2.2.3 Blood oxygen level–dependent fMRI 205
7.3 fMRI and perception of odorous compounds 208
7.3.1 Anatomy and organisation of the olfactory system 209
7.3.1.1 Olfactory receptors, olfactory nerves and
olfactory bulb 210
7.3.1.2 Olfactory tract and primary olfactory cortex 211
7.3.2 fMRI studies of the primary olfactory cortex 213
7.3.2.1 Secondary olfactory regions 215
7.3.2.2 Orbitofrontal cortex 216
7.3.2.3 The role of the amygdala in hedonic
processing of odours 218
7.3.2.4 Other brain regions 220
7.3.2.5 Cingulate 220
7.3.2.6 Cerebellum 220
7.3.2.7 Imagination of odours and memory 221
7.4 Interaction between olfaction and other sensory modalities 221
7.4.1 Transduction of pheromone-like compounds in humans 222
7.5 Conclusion 222
8 Flavor interactions at the sensory level 228
RUSSELL S. J. KEAST, PAMELA H. DALTON
and PAUL A. S. BRESLIN
8.1 Introduction 228
8.2 The psychophysical curve: physical intensity
vs. perceived intensity 228
8.3 Attributes of sensory modalities 234
8.3.1 Quality 234
8.3.2 Intensity 234
8.3.3 Temporal pattern 235
8.3.4 Spatial pattern 236
CONTENTS ix
8.4 Adaptation 237
8.5 Four levels of flavor interactions 237
8.5.1 Physiochemical interactions 237
8.5.2 Mechanical/structural interactions 238
8.5.3 Oral and nasal peripheral interactions 238
8.5.4 Central cognitive interactions 239
8.6 Intramodal interactions 239
8.6.1 Taste 239
8.6.1.1 Single-quality interactions 240
8.6.1.2 Multiple-quality interactions 241
8.6.2 Odor 241
8.6.2.1 Odor interactions 242
8.6.3 Somatosensations are components of flavor 242
8.6.3.1 Chemesthesis: irritation 243
8.7 Texture 244
8.7.1 Visual texture 245
8.7.2 Auditory texture 245
8.7.3 Tactile texture 245
8.8 Interactions between modalities 246
8.8.1 Interactions of orosensory chemesthesis, tactile
sensations and taste 246
8.8.2 Interactions of odor and somatosensations 247
8.8.3 Interactions of odor and taste 247
8.9 Sources of error in sensory research 248
8.9.1 Individual variation 248
8.9.2 Experimental protocol 248
8.9.3 Choice of flavor-active compound 249
8.9.4 Psychophysical function of a compound 249
8.9.5 Method of rating 249
8.10 Practical implications for flavor 249
9 Psychological processes in flavour perception 256
JOHN PRESCOTT
9.1 Flavour as sensory integration 256
9.2 Qualitative and psychophysical evidence for odour–taste
integration 257
9.2.1 Taste properties of odours 258
9.2.2 How do taste-related odour qualities develop? 258
9.2.3 How ‘real’ are smelled taste qualities? 259
9.2.4 Influence of smelled taste qualities on the perception
of tastes 261
x CONTENTS
9.3 Smelled taste qualities and taste modification as indicators
of flavour formation 263
9.3.1 The role of spatial and temporal factors in
odour–taste integration 266
9.4 Cognitive processes in the development of flavour perception 267
9.4.1 Taste modification by odours: a rating effect? 267
9.4.2 Taste modification by odours as a function of
perceptual strategy 268
9.4.3 Analysis and synthesis in the perception of flavour 269
9.4.4 Investigating cognitive processes in flavour perception 270
9.5 Implications and future directions 273
9.6 Conclusion 274
Index 279
CONTENTS xi
Contributors
Anthony A. Blake Firmenich SA, Building 41, Rue de la Bergere 7,
1217 Meyrin 2, Geneva, Switzerland
Paul A. S. Breslin Monell Chemical Senses Center, 3500 Market
Street, Philadelphia, PA 19104-3308, USA
Loıc Briand INRA, Unite Rech 477, Domaine de Vilvert,
F-78352 Jouy-en-Josas, France
Pamela H. Dalton Monell Chemical Senses Center, 3500 Market
Street, Philadelphia, PA 19104-3308, USA
Rene de Wijk Wageningen Centre for Food Sciences, Diedenwes
20, 6700 Av Wageningen, The Netherlands
Barry G. Green The John B. Pierce Laboratory, Yale University
School of Medicine, New Haven, CT 06510, USA
Joanne Hort Division of Food Sciences, University of
Nottingham, Sutton Bonington Campus,
Loughborough LE12 5RD, UK
Russell S. J. Keast Monell Chemical Senses Center, 3500 Market
Street, Philadelphia, PA 19104-3308, USA
Birgit Kettenmann Department of Radiology, Virginia Commonwealth
University Health System, 401 N. 12th Street,
Richmond, VA 23298, USA
Gerd G. Kobal Sensory Research, WSA Philip Morris USA, 615
Maury Street, Richmond, VA 23224, USA
Xia Li Monell Chemical Senses Center, 3500 Market
Street, Philadelphia, PA 19104-3308, USA
Jean-Claude Pernollet INRA, Unite Rech 477, Domaine de Vilvert,
F-78352 Jouy-en-Josas, France
John J. Prescott University of Otago, Sensory Science Research
Centre, PO Box 56, Dunedin, New Zealand
Jon F. Prinz Wageningen Centre for Food Sciences, Diedenweg
20, 6700 AN Wageningen, The Netherlands
Nancy E. Rawson Monell Chemical Senses Center, 3500 Market
Street, Philadelphia, PA19104-3308, USA
Deborah D. Roberts Food and Flavor Science Consulting LLC, 17632
Ailanthus Dr, Chesterfield, MO 63005, USA
Andrew J. Taylor Division of Food Sciences, University of
Nottingham, Sutton Bonington Campus,
Loughborough LE12 5RD, UK
Martin Wiesmann Department of Neuroradiology, University of
Munich, 15 Marchionin Streeet, Munich 81377,
Germany
CONTRIBUTORS xiii
Preface
Unlike other human senses, the exact mechanisms that lead to our perceptionof flavour have not yet been elucidated. There are many reasons for our lack ofunderstanding.
First, we now recognise that flavour perception involves a wide range ofstimuli (also called modalities). The simple notion that flavour is the result ofthe well known taste and aroma modalities has been replaced by a realisationthat it is a multi-modal phenomenon. Identifying the modalities involved ineach food is difficult. Within each modality, there can be many contributingfactors and a corresponding array of sensors. Aroma is the most complex, witharound 2,500 odorous chemical compounds found in food and around 1000receptors. Taste is somewhat simpler, with 5 basic components (salt, sour,sweet, acid and umami) and a corresponding number of receptors. Besidestaste and aroma, attributes like colour, appearance and sound (e.g. the crispycrunch of some foods) are relatively easy to define. The inner surface of themouth also contains many receptors that sense stimuli like pain and heat, aswell as mouthfeel attributes like creamy, slimy and gritty – but these senses arenot as well defined as the taste and aroma receptors.
Second, the chemical compounds and physical structures that activate theflavour sensors change as food is eaten. For instance, the initial crisp nature ofa biscuit is replaced by a sticky bolus as the food material becomes hydratedduring eating, changing the textural attribute. With wine, the initial aromasniffed from the glass is replaced by a burst of flavour as the wine is sipped,and that in turn is followed by complex sensations involving terms like fruity,plummy, vanilla, smoky, etc. Measurements of the changes in stimuli withtime are therefore essential to an understanding of the relationship betweenstimuli and perception.
Third, there is a wealth of anecdotal evidence that the individual modalitiesinteract in a complex way and that the overall perception of flavour is not asimple additive effect. Whether these interactions are the result of ‘hardwiring’ of the brain during development or whether they are learned throughexperience is not entirely clear.
Fourth, the sense of flavour is not as well developed as other senses likespeech, music or vision, and we lack a notation (or language) that canadequately describe flavour in terms of the stimuli that create it.
Despite these gaps in our understanding, huge progress has been made inmany of the above fields and some examples are listed below.
. Measuring flavour stimuli in vivo
. Molecular biology studies of the olfactory and gustatory receptors
. Prediction of olfactory receptor structure and function from primary aminoacid sequences
. Neurobiological studies
. Functional MRI
. Multimodal perception
If we are to make further progress in understanding the origins of flavourperception, it is clear that we need to consider the whole process – the releaseof flavour chemicals in mouth, the transport processes to the receptors, thespecificity and characteristics of the receptors, the transduction mechanismsand the subsequent processing of signals locally and at higher centres in thebrain. The use of neurobiological techniques and functional MRI can enable usto visualise the effects of flavours on brain function and, coupled with psy-chological and sensory studies, we can follow each stage of the flavourperception process.
Most of the progress to date has been in deepening our understanding ofthese individual areas. In order to progress further, approaches integratingthese areas will certainly be fruitful. However, a good part of the researchrelated to flavour perception is presented at separate scientific meetings ofchemists, biologists, psychologists or physiologists and it is not easy to gain anunderstanding of the whole field from a single source. This book aims toencapsulate current knowledge in chapters covering the key stages of flavourperception, and to produce a source of information that will be useful to peopleworking in the flavour field, whether they be in academia or in the food/flavour industry.
During the writing of these chapters, we also organised a symposium on thesubject, which provided an opportunity for the authors to meet and yieldedmuch useful information which has been incorporated into some of the chap-ters. We would like to acknowledge the support of the United States Depart-ment of Agriculture Cooperative State Research, Education and ExtensionService (USDA 2004-35503-14178) and the commercial sponsors (NestleSA, Firmenich SA, Kraft Foods, International Flavors and Fragrances Inc.,Proctor and Gamble, Givaudan Flavors Corporation, and Symrise) for theirfinancial support.
Flavour research is of interest not only on account of its hedonistic implica-tions but because it is one of the factors that limits the acceptance of more
xvi PREFACE
nutritious foods and the enjoyment of food by the ageing population. A basicunderstanding of the origins of flavour perception would therefore have manyuses in the food industry.
AJ TaylorDD Roberts
PREFACE xvii
B
G
R
Opponent neurons
Dim
Bright
−ve greenish
+ve reddish
−ve bluish
+ve yellowish
R-G = C1
G-B = C2
B-R = C3
C2-C3
Receptor cells
Figure 1.10 Representation of Opponent Colour theory.
5R
5YR
5Y
5GY
5G5BG
5B
5PB
5P
5RP
Value
Hue
Chroma
white
9
8
7
6
4
2
1
black
Figure 1.11 Munsell Colour system. (Picture courtesy of HunterLAB.)
Figure 1.12 HunterLAB colour scale showing the white/black axis (L, vertical), the red/green axis (þa
to �a) and the yellow/blue axis (þb to �b). (Picture courtesy of HunterLAB.)
1 Measuring proximal stimuli involved in
flavour perception
Andrew J. Taylor and Joanne Hort
1.1 Factors influencing flavour perception
Humans eat food both for nutritional benefit and for pleasure. The processes inboth cases involve the interaction of food components with specific receptorsin the body, which leads to the stimulation of a wide range of sensing systemsin humans. While the effects of nutritional stimuli on our behaviour are largelyunderstood (e.g. the effect of carbohydrate consumption and hydrolysis, bloodglucose levels and their consequences for attention and appetite), the samecannot be said for flavour stimuli. From our experience, and from formaldictionary definitions of ‘flavour’, it is clear that several senses (often calledmodalities) are involved in flavour perception although a complete list ofsenses/stimuli is rarely given. It is likely that the stimuli involved are highlydependent on the food type. For chocolate, the initial mechanical properties ofthe solid chocolate and the ‘mouthfeel’ of the melted chocolate, coupled withthe characteristic chocolate taste and aroma, are probably the key modalitiesinvolved in flavour perception and in that order of importance. With snackfoods, crispness is essential while salt and flavour create the subsequentsavoury flavour, whereas with wine it is most likely an equal combination ofaroma, taste and mouthfeel. Therefore, we need to identify the key sensoryattributes for each food product and not forget that other senses can affect ourperception. For example, the discoloration of chocolate due to surface fatcrystallisation (so-called bloom) causes little change to the aroma, taste ormouthfeel modalities but reduces enjoyment of the product through a visualstimulus.
1.1.1 Perception of ‘taste’ and its location
Typical flavour definitions include the taste and aroma modalities, usuallywith some mention of ‘mouthfeel’, broadly described as the stimuli that occurin the mouth during eating. These stimuli and their detection are technicallydescribed as chemesthesis and are discussed in detail in Chapter 5. Thereare also definition difficulties due to the imprecise nature of the words thatwe use to describe food flavour. In English , the word ‘taste’ means not onlythe gustatory mechanism found on the tongue for the primary salt, acid, sweet
and bitter modalities but it also describes our overall impression of a food as inthe phrase ‘that tastes good’. Therefore, although flavour is perceived byreceptors in the eye, tongue, nose and mouth lining, the brain interprets theoverall sensation as occurring in the mouth. Green (2001) has provided anexplanation for this observation by suggesting that the brain ‘localises’ all thesensory information into the mouth, so we associate that information withthe food we consume in the same way we use sight and touch to localise apoint on our bodies. Prescott (1999) talks about the ‘illusion’ of flavourappearing to originate in the mouth and suggests that, since the mouth actsas a gateway to the gut, the sense of flavour determines what foods can enter tokeep us safe. Because we localise flavour perception in the mouth, it is notimmediately obvious which stimuli are involved in flavour perception andhow we combine, vocalise and locate these signals. The aim of this chapter isto study the stimuli involved in flavour perception to set the scene for thefollowing chapters. Subsequent chapters will consider sensing of the stimuliand how the signals produced are processed to provide us with a ‘perceptualconstruct’ of that particular flavour.
1.1.2 Multimodal nature of flavour perception
The definitions of flavour listed above are those typically found in scientificliterature associated with flavour research but, increasingly, the conceptsdeveloped in other scientific disciplines are being applied, leading to a re-assessment of the best definition for flavour. Perception itself has been studiedextensively, initially to understand the mechanisms of visual and auditoryperception but later to understand how these individual senses combine andinteract, leading to the idea of multimodal and crossmodal perception, respect-ively. In brief, multimodal refers to the fact that several senses (modalities) areinvolved in flavour perception, and crossmodal indicates that one modality caninteract with another to modify the perception. There is now evidence fromphysiological, neural and functional magnetic resonance imaging (fMRI)studies of brain (see Chapter 7) that leads to a consensus view, namely, flavourperception is no different from other senses and involves both multi- andcrossmodal activities. Why has it taken so long to apply this apparentlyobvious concept to flavour perception? First, it seems that early anatomicalstudies on neural pathways from the aroma and taste receptors showed thatthey entered the brain in different places and this was initially interpreted asproof that the two stimuli were independent. A second reason is that earlywork on multimodal perception proposed that modalities were hierarchical.Vision was considered the most important modality and the hierarchicalscheme proposed that vision would always be the dominant sense and wouldoverride the input from other modalities. This view is no longer held, as thereare many examples that contradict the purely hierarchical idea. A pertinent
2 FLAVOR PERCEPTION
example for this book is that a food, however visually appealing, will not beconsumed if it emits a foul odour. In this particular case, odour, not vision, isthe dominant modality. A third reason for not considering the multimodalapproach is that some interactions noted in the literature could be due tophysicochemical interactions or artefacts of the sensory analysis methodsused, rather than any cognitive, multimodal effect. Here, we define a cognitiveeffect as any interaction that occurs in signal processing after stimulation ofthe receptors. For instance, enhancement of aroma by sweeteners (Duran &Costell, 1999) could be due to the presence of sugar at concentrations from 5%to 10% in the samples that might increase aroma release through the well-known physicochemical interaction of solutes (see Taylor, 1998 for review).Thus the observed sensory behaviour could be due to either the physicochem-ical effect or interaction at the cognitive level. Recent work has shown that lowconcentrations (< 10%) of solutes like sugar do not affect the in vivo release ofaromas significantly (unless aromas bind to the solute). In these former cases,the effect is not due to an increased aroma signal being transported to theolfactory receptors (ORs: Breslin, 2001; Davidson et al., 1999; Friel et al.,2000). Lastly, because of the complexity of flavour, researchers have adoptedthe classical scientific reductionist approach that seeks to break down theproblem into its component parts. This has led to the development of subfieldsin flavour research such as flavour chemistry, sensory analysis, flavour releaseand molecular biology of taste and aroma receptors. While this simplifies thesystems so they can be investigated, it is imperative that researchers do notlose sight of the whole picture and realise that an understanding of flavourperception requires an integration of all the modalities using a multidisciplin-ary approach.
Some authors have questioned whether the interactions observed betweenmodalities such as taste and aroma are artefacts of the sensory analysis processitself. In some sensory experiments, panellists may have limited attributes inwhich to score the samples and there is a notion that panellists may ‘dump’ anyother differences into that (inappropriate) category. For example, when panel-lists are asked to assess a system containing aroma and sugar for sweetness,they may inadvertently assign a change in aroma perception to the sweetnesscategory. Prescott (1999) has discussed the cause of taste–aroma interactionsand has discounted the idea of dumping. He does point out that interactions areseen only if the panellist is presented with the sample in a particular way and ifthere is already some prior experience of that taste–aroma pairing (so-calledcongruency). However, the interactions have been noted by many differentresearch groups across the world using a wide range of sensory methods andthere is general agreement that the effects observed are real. The general areaof crossmodal flavour interactions has been reviewed (Duran & Costell, 1999).Neurobiological studies in animals have demonstrated that there are connec-tions between the taste and aroma systems, and specific neurons that require
MEASURING PROXIMAL STIMULI INVOLVED IN FLAVOUR PERCEPTION 3
a taste and aroma signal before they are activated have been identified (Rolls& Baylis, 1994). All these results are relatively new and, taken together,provide a consensus platform from which our understanding of flavour per-ception can move forward. Further discussion of interactions between flavourmodalities that cause sensory differences can be found in Chapters 8 and 9.
1.1.3 Distal and proximal stimuli
If the notion of multimodal flavour perception is to be studied, we need tobe able to identify and measure the stimuli involved, but again we need to bearin mind some principles from other scientific disciplines. The most importantis the concept of distal and proximal stimuli, which is demonstrated in Figure1.1. The distal stimulus is defined as the whole object (in Figure 1.1, a skull),but the perception of that object by vision involves sensing only part of theobject, and the image is very small (a few mm) and is expressed upside downon the retina of the eye. Although the eye sees only the front, side or back ofthe skull at any one time, our familiarity with this object means that we can fillin the missing parts and we imagine that the whole skull is present. Forunfamiliar objects, we tend to rotate them to obtain the full distal signal;thereafter, only a partial signal is necessary to recognise the object.
1.2 Aroma
How does the distal/proximal concept apply to the components that make upthe sense of flavour? Figure 1.2 outlines the situation for aroma perception.The relationships between distal stimuli, proximal stimuli and aroma percep-tion are shown along with the corresponding compositions and methodologiesfor measuring the composition at the different stages. Thus the aroma com-position of the sample is the distal stimulus, measured by conventional
Distalstimulus
Proximalstimulus Perception
Skull
Human eye
Figure 1.1 Representation of a distal stimulus (a skull), the signal received by the receptor (the proximal
stimulus on a small scale – inverted image of the front of a skull) and the perceptual response (recognition of a
skull from the information provided).
4 FLAVOR PERCEPTION
quantitative extraction and gas chromatography–mass spectrometry (GC–MS).Aroma can be sensed by the orthonasal route (i.e. sniffed through the nostrils),in which case the volatile content in the gas phase above a food (the so-calledheadspace) is a good indicator of the proximal stimulus that reaches the ORs(see Section 1.2.1). Alternatively, aroma compounds can reach the ORs via thethroat during and after the mastication process, the so-called retronasal route.Mastication causes significant changes to the food in terms of surface area,hydration and time in mouth, and these factors in turn affect the mass transferprocesses involved in transporting aroma compounds from the food to the gasphase in the mouth and then to the ORs in the nose (van Ruth & Roozen,2002). Thus the retronasal proximal stimulus may be very different from thedistal stimulus of a food. The effects of oral physiology on flavour release arediscussed in Chapter 2.
1.2.1 Measurement of orthonasal proximal stimuli
When considering how to measure the orthonasal aroma stimulus, it is neces-sary to consider the various types of food and our typical ways of consumingthem. If we take an example of a packaged food (e.g. roasted peanuts in asealed bag), the physical conditions inside the bag create an equilibriumheadspace, i.e. the aroma compounds are partitioned between the solid peanutphase and the gas phase at an equilibrium (and constant) value. The equilib-rium value will depend on temperature but is one of the parameters widelyused in aroma analysis of foods as it provides a fundamental value (theair–food partition) that gives an indication of the amount of each volatilecompound above a food as well as of the potential flavour release duringeating when incorporated in mathematical models (see Linforth, 2002 for a
Distal stimulusProximalstimulus Perception
Measured byextraction and GC−MS
Measured byAPCI
Sensorymeasurements
Flavourcomposition in
food
Aroma and tasteprofile atreceptors
Ortho-, retro-nasal signals
Combination oftaste, aroma
and mouthfeelsignals in brain
StaticheadspaceOrthonasal
signal
Figure 1.2 Relationship between distal and proximal stimuli (top row), flavour-sampling methods (middle
row) and flavour analyses (bottom row).
MEASURING PROXIMAL STIMULI INVOLVED IN FLAVOUR PERCEPTION 5
review of flavour release models). When a bag of peanuts is opened close tothe nose, the first stimulus is very close to the equilibrium headspace but thereis rapid dilution of that headspace as air is drawn into the packet and theconcentrations of individual compounds in the headspace change with dilu-tion. This change in headspace concentration with dilution can be measuredusing the technique of dynamic headspace dilution where the initial equilib-rium between the food and the headspace is disturbed by a flow of gas throughthe headspace (Taylor, 1998). The technique and the effect of dilution onaroma compounds has been described previously (Marin et al., 1999, 2000).The change in headspace composition with time can sometimes be demon-strated with bags of peanuts by opening the packet very close to the nose andimmediately taking a deep sniff. Often, a strong sulphur note can be detectedbut this dissipates almost immediately as the sulphur compound has a very lowpartition value and the concentration in the headspace is not replenishedsufficiently quickly for it to be detected subsequently. Another example ofthe change of aroma composition with time is provided by the process of winetasting. Although individual wine tasters adopt slightly different styles ofsampling the wine, they all employ the routine of sniffing the ‘bouquet’ ofthe wine (i.e. an initial assessment of odour via the orthonasal route) followedby complex mouth movements (some seem to chew the wine), which maxi-mise aroma release and transport the aroma compounds to the OR via theretronasal route. This manipulation of flavour release allows the wine taster toproduce a sequence of verbal descriptors, presumably as a result of thesequential delivery of flavour stimuli.
In the case of both peanut and wine aromas, it is clear that the proximalstimuli reaching the ORs are very different from the distal flavour compositionof peanuts or wine, and this may be one of the reasons why it has proveddifficult to relate flavour perception to flavour composition in the past.Improved devices to mimic orthonasal stimuli are needed; one such device isthe gas–liquid interfacial mass-transfer (GLIM) cell proposed by Parker(Parker & Marin, 2000), in which the dynamic release of aroma can bemeasured as the gas and liquid phases are diluted.
1.2.2 Measurement of retronasal proximal stimuli
The processes involved in delivering the aroma stimuli to the ORs have beenreviewed recently (Taylor, 2002). Measuring the stimuli is ideally carried outas close to the receptors as possible. Given ethical considerations and the factthat sampling air at the ORs requires invasive (and potentially painful)methods, it is more convenient to sample air as it exits the nostril. Earlywork from our laboratory established that release profiles could be obtainedby sequential trapping of volatile compounds on Tenax, but the process wasvery time-consuming (Ingham et al., 1995). The development of online mass
6 FLAVOR PERCEPTION
spectroscopy (MS) techniques like atmospheric pressure chemical ionisation(APCI) or proton transfer reaction (PTR) to monitor aroma release on a breath-by-breath basis has provided new opportunities for studying aroma release invivo. The techniques have been optimised for sensitivity (typically nanolitresof aroma per litre of air; ppbv) as well as coping with the humidity of exhaledair and the difficulty of ionising a wide range of volatile compounds simultan-eously ( see Taylor et al., 2000 for review of APCI and van Ruth et al., 2003for PTR). Although both APCI and PTR have contributed much to our under-standing of aroma release, both techniques have limitations in sensitivity andselectivity. The odour thresholds for some volatile compounds lie in the parts-per-trillion (ppt) range (picolitres of aroma vapour per litre of air), and thus thetypical ppbv sensitivity of the MS techniques is inadequate. Some workershave achieved very low sensitivity but mainly with compounds possessing ionmasses away from the chemical noise associated with chemical ionisationtechniques or with long dwell times that do not allow breath-by-breath analy-sis. Thus the presence of chemical warfare agents in a smokestack was reportedin the low ppt range (Ketkar et al., 1991) because the ion mass occurred outsidethe chemical noise range (giving an excellent signal–noise ratio) and becausethe ion could be monitored for relatively long periods of time. For aromacompounds with molecular weights (and ion masses) in the 45–200Da range,ppbv is the typical detection threshold although certain compounds can befollowed down to 100 ppt. Secondly, the complex composition of many aromasresults in the presence of several compounds with the same molecular weight(isobaric compounds). These cannot be differentiated by the APCI or PTRtechniques that only resolve on a unit mass basis. Although some selectivitycan be gained by using MSn (where the original ion is fragmented and theresulting ions analysed using either a tandem MS system or an ion trapinstrument), compounds like terpenes create problems as they each haveindividual aromas but are all found at a few common ion masses.
One of the future drivers for improved real-time MS of aroma release maybe developments in MS related to the study of metabolomics. There are manysimilarities between this area of research and the analysis of flavour com-pounds. Although there are many definitions of metabolomics, the underpin-ning idea is the ability to analyse a wide range of compounds of differentchemical classes at varying concentrations in a rapid and quantitative mannerwithout creating artefacts or losing labile compounds. Such an analysis haslong been the goal of flavour chemists. New developments in MS include theuse of alternative ionisation methods. For example, photoionisation (PI) is asoft ionisation that has the potential to ionise molecules in a mixture select-ively by using different wavelengths and energies of the light. Another devel-opment is the availability of time of flight (TOF) mass analysers that are wellsuited to the simultaneous analysis of many different ions and also giveaccurate mass data. Increasingly, hybrid techniques are being proposed such
MEASURING PROXIMAL STIMULI INVOLVED IN FLAVOUR PERCEPTION 7
as the use of PI to ionise molecules, followed by an ion trap to select ionmasses and perform fragmentation of the molecule to provide more infor-mation on its identity (the MSn technique), followed by detection in a TOFanalyser. The development and manufacture of such machines will be drivenby other, more lucrative research fields than flavour analysis, but when suchmachines become available, there will undoubtedly be a move to adapt themfor the analysis of flavours as well as metabolic profiles. The outcome will beimproved detection and identification of the proximal flavour stimuli, leadingto a better understanding of the concentration change in proximal aromastimuli with time.
Both APCI and PTR can be used for monitoring the aroma stimuli frommodel in vitro as well as in vivo systems. In vivo sampling has the advantagethat it measures the actual aroma signal in people as they consume foodsamples. The disadvantages are that person-to-person variation is high dueto variability in their mastication behaviour and in their physiology, and bothfactors cause significant differences in the aroma release profile observed.However, release from an individual is usually consistent (Haring, 1990).Thus if the aim of an experiment is to test the effect of a different foodcomposition on aroma release, it is best to use one or two individuals whorelease aroma consistently. This will identify the compositional factors influ-encing aroma release without confusing the issue with the inherent variation ina group of people.
The amounts of volatile compounds released in vivo are also small asthe food sample is typically limited to about 10 g. One way to overcome thehuman variation in eating behaviour and the small food sample size is to builda model mouth in which larger samples can be subjected to controlled ‘masti-cation’ using the information available on oral physiology (e.g. salivaflow, volume of mouth, air flow). Various devices have been described andall give reproducible results. The question that remains unanswered for manyof the model mouths, however, is whether the aroma signal they producerelates to the signal seen in vivo. Given the variability of humans, this mayseem an impossible task but panellists can be instructed to eat to a certainpattern and individuals with atypical release can be screened out of experi-ments. There are also some food systems that produce recognisable patternsof aroma release in vivo and some validation of model mouths has beenattempted. The retronasal aroma simulator (RAS) was tested using APCI tomonitor its release pattern for a variety of real foods; these were then com-pared with human release patterns and a good correlation was found (Deibleret al., 2001). The intensity of volatile compounds released from the RAS wasmuch higher due to the larger amount of sample used and the air flow was akey parameter in determining the release profile. Van Ruth has recentlyreported the same sort of experiment to validate the model mouth developedat Wageningen (van Ruth et al., 2002). Rabe has described a model mouth
8 FLAVOR PERCEPTION