Odor-active constituents in fresh pineapple (Ananas comosus [L.] Merr.) by quantitative and sensory evaluation

By application of aroma extract dilution analysis (AEDA) to an aroma distillate prepared from fresh pineapple using solvent-assisted flavor evaporation (SAFE), 29 odor-active compounds were detected in the flavor dilution (FD) factor range of 2 to 4,096. Quantitative measurements performed by stable isotope dilution assays (SIDA) and a calculation of odor activity values (OAVs) of 12 selected odorants revealed the following compounds as key odorants in fresh pineapple flavor: 4-hydroxy-2,5-dimethyl-3(2H)-furanone (HDF; sweet, pineapple-like, caramel-like), ethyl 2-methylpropanoate (fruity), ethyl 2-methylbutanoate (fruity) followed by methyl 2-methylbutanoate (fruity, apple-like) and 1-(E,Z)-3,5-undecatriene (fresh, pineapple-like). A mixture of these 12 odorants in concentrations equal to those in the fresh pineapple resulted in an odor profile similar to that of the fresh juice. Furthermore, the results of omission tests using the model mixture showed that HDF and ethyl 2-methylbutanoate are character impact odorants in fresh pineapple.     

The Role of Perceptual and Structural Similarity in Cross-adaptation

Cross-adaptation, the decrease in sensitivity to one odorantfollowing exposure to a different odorant, is affected by odorantsimilarity, both perceptual and structural, but the preciserelationship is obscure. The present series of studies was designedto explore various aspects of perceptual and structural similarityas they relate to cross-adaptation. In Experiment 1, cross-adaptationwas assessed between androstenone and five odorants that sharea common urinous note with androstenone, but retain unique perceptualcharacteristics; only the compound judged most perceptuallysimilar to androstenone cross-adapted it. In Experiment 2, odorantsboth perceptually and structurally similar (androstenone andandrostanone) displayed significant, mutual cross-adaptation.Furthermore, magnitude estimates for androstanone were significantlyreduced following exposure to 3-methylidene-5a-androstane (3M5A),a structurally similar, perceptually odorless compound. Thisfinding appears to be the first demonstration that an odorlesscompound can affect, via cross-adaptation, the perception ofan odorous compound. Finally, in Experiment 3, significant,asymmetric cross-adaptation was observed between compounds thatare perceptually and structurally dissimilar (4-cyclohexylcyclohexanone[4-CHCH] and androstenone). These findings indicate that therole of similarity in cross-adaptation is difficult to quantifyand emphasize the numerous odorant characteristics that canaffect cross-adaptation. Chem. Senses 21: 223–237, 1996.     

Configurational and Elemental Odor Mixture Perception Can Arise from Local Inhibition

Contrast enhancement via lateral inhibitory circuits is a common mechanism in sensory systems. We here employ a computational model to show that, in addition to shaping experimentally observed molecular receptive fields in the olfactory bulb, functionally lateral inhibitory circuits can also mediate the elemental and configurational properties of odor mixture perception. To the extent that odor perception can be predicted by slow-timescale neural activation patterns in the olfactory bulb, and to the extent that interglomerular inhibitory projections map onto a space of odorant similarity, the model shows that these inhibitory processes in the olfactory bulb suffice to generate the behaviorally observed inverse relationship between two odorants' perceptual similarities and the perceptual similarities between either of these same odorants and their binary mixture.     

Mutual cross-adaptation of the volatile steroid androstenone and a non-steroid perceptual analog

Self-and cross-adaptation are believed to result from stimulationof the same olfactory sensory channels. These adaptation phenomenawere studied after exposures to 5-androst-16-en-3-one (androstenone)and a synthetic perceptual analog (DMCMC), viz. a racemic mixtureof the isomers 4(R)-(4',4'-dimethyl-cyclohexyl)-2(R)-methylcyclohexanoneand 4(S)-(4',4'-dimethylcyclohexyl)-2(S)-methylcyclohexanone.In Experiment 1, six subjects very sensitive to androstenonereceived four randomized sequences of six concentrations offour ordants (androstenone, DMCMC, amyl acetate, and Galaxolide*;plus blanks) before and following adaptation to either androstenoneor DMCMC. Exposure to each odorant resulted in self-adaptation.Measures of stimulus intensity and identification thresholdrevealed reciprocal cross-adaptation between androstenone andDMCMC, but no cross-adaptation to amyl acetate or Galaxolide.The degree of cross-adaptation was asymmetric; adaptation toDMCMC resulted in more complete adaptation to androstenone thanvice versa. This asymmetry was apparently due to intensity differences;when stimuli were matched for intensity, the asymmetry disappeared(Experiment 2). These results demonstrate cross-adaptation forqualitatively similar, but not dissimilar, odors and suggestthat androstenone and its perceptual analog DMCMC share thesame sensory channels.     

Semantic, typicality and odor representation: a cross-cultural study

This study investigated odor-category organization in threecultures by evaluating (i) the relationship between linguisticand perceptual categorization and (ii) the existence of an internalstructure of odor categories. In the first experiment, threegroups of 30 participants from American, French and Vietnamesecultures performed a sorting task. The first group sorted 40odorants on the basis of odor similarity, the second group sorted40 odor names on the basis of name similarity and the last groupsorted 40 odor names on the basis of imagined odor similarity.Results showed that odor categorization was based on perceptualor conceptual similarity and was in part independent of wordand imagined categorizations. In the second experiment, anothergroup of 30 participants from each culture rated the typicalityof the odorants for 11 odor categories. Results showed thatsome odorants were rated as more typical than others. Moreover,the typicality gradient predicted the odor space obtained inthe odor sorting task in a consensual way among the three cultures.These results suggest that, as for other categories, odor categoriesare based on perceptual similarities rather than on semanticcues. Moreover odor-category structure might have a core representationwhich might be common to different cultures with boundarieswhich might be more culturally dependent.     

Enantioselectivity of the musk odor sensation

This brief review, the summary of a talk at the Symposium on Biological Chirality 2000 in Szeged, Hungary, illustrates what chiral recognition tells us about the molecular parameters of the musk odor sensation. While the enantioselectivity of odor perception is strong evidence for the key role of proteinogenic receptors in the molecular mechanism of olfaction, the quantitative and qualitative odor differences of enantiomers are often not very pronounced, as in the case of muscone (17/26). In those cases, however, where there is strong enantiodiscrimination, we find most intense musk odorants with very low odor thresholds, such as (-)-(12R)-12-methyl-9-oxa-14-tetradecanolide (35), (12R;9Z)-12-methyl-14-tetradec-9-enolide [(R)-Nirvanolide, 38], and (-)-(4S;7R)-1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylcyclopenta[g]-2-benzopyran [(-)-(4S;7R)-Galaxolide, 57], the latter being rather rigid. We thus can assume the geometry of the musk receptor to be fairly complementary to these compounds, which therefore can serve as templates for the design of new musk odorants.     

Dissociable codes of odor quality and odorant structure in human piriform cortex

The relationship between odorant structure and odor quality has been a focus of olfactory research for 100 years, although no systematic correlations are yet apparent. Animal studies suggest that topographical representations of odorant structure in olfactory bulb form the perceptual basis of odor quality. Whether central olfactory regions are similarly organized is unclear. Using an olfactory version of fMRI cross-adaptation, we measured neural responses in primary olfactory (piriform) cortex as subjects smelled pairs of odorants systematically differing in quality and molecular functional group (as one critical attribute of odorant structure). Our results indicate a double dissociation in piriform cortex, whereby posterior regions encode quality (but not structure) and anterior regions encode structure (but not quality). The presence of structure-based codes suggests fidelity of sensory information arising from olfactory bulb. In turn, quality-based codes are independent of any simple structural configuration, implying that synthetic mechanisms may underlie our experience of smell.     

Cross-adaptation between olfactory responses induced by two subgroups of odorant molecules

It has long been believed that vertebrate olfactory signal transduction is mediated by independent multiple pathways (using cAMP and InsP3 as second messengers). However, the dual presence of parallel pathways in the olfactory receptor cell is still controversial, mainly because of the lack of information regarding the single-cell response induced by odorants that have been shown to produce InsP3 exclusively (but not cAMP) in the olfactory cilia. In this study, we recorded activities of transduction channels of single olfactory receptor cells to InsP3-producing odorants. When the membrane potential was held at -54 mV, application of InsP3-producing odorants to the ciliary region caused an inward current. The reversal potential was 0 +/- 7 mV (mean +/- SD, n = 10). Actually, InsP3-producing odorants generated responses in a smaller fraction of cells (lilial, 3.4%; lyral, 1.7%) than the cAMP-producing odorant (cineole, 26%). But, fundamental properties of responses were surprisingly homologous; namely, spatial distribution of the sensitivity, waveforms, I-V relation, and reversal potential, dose dependence, time integration of stimulus period, adaptation, and recovery. By applying both types of odorants alternatively to the same cell, furthermore, we observed cells to exhibit symmetrical cross-adaptation. It seems likely that even with odorants with different modalities adaptation occurs completely depending on the amount of current flow. The data will also provide evidence showing that olfactory response generation and adaptation are regulated by a uniform mechanism for a wide variety of odorants.     

Analysis of Odor Discrimination by Multidimensional Scaling at Different Temperatures

Discrimination of odorants by the turtle olfactory bulb at 25°and 37°C was examined by the cross-adaptation techniqueand analysed by multidimensional scaling. Analysis by multidimensionalscaling suggests that at 25°C odorants are grouped accordingto their odor qualities in the turtle olfactory system. At 37°C,the cluster formation of odorants, which have a similar odorquality, such as minty and floral alcohol odorants and molecularstructure, in the plot of multidimensional scaling was poor,indicating that the ability of odor grouping according to theirodor qualities was low at 37°C. Chem. Senses 20: 565–571,1995.     

The influence of training on chemosensory event-related potentials and interactions between the olfactory and trigeminal systems

It is not possible to accurately predict the perceptual response to odorants and odorant mixtures without understanding patterns of suppression and facilitation that result from interactions between the olfactory and trigeminal systems. The current study extends previous findings by exploring the effect of intensive training on the interaction between these systems and also by using a different mixed chemosensory stimulus to examine whether the principles established in earlier studies generalize to different odorants. Stimuli were chosen so as to selectively activate the olfactory (H2S) and trigeminal (CO2) nerves. In addition, linalool was included as a stimulus that activated both systems. Thirty-five participants (19 men, 16 women) rated the intensity of each stimulus when presented both alone and in binary mixtures (linalool + H2S, and linalool + CO2). Chemosensory event-related potentials were obtained from three recording positions. Analysis of intensity ratings showed that linalool was significantly less intense than the other stimuli when presented alone. In binary mixtures, H2S was strongly suppressed by linalool. One week of intensive odor training produced significant and specific reductions in the intensity of linalool and H2S, both alone and in their mixture. Training with a different odor (champignol) had no effect. Chemosensory event-related potential data confirmed previous findings showing changes in topographical distribution that reflected the degree of trigeminal activity. Binary mixtures generally produced larger amplitudes than single stimuli. Latencies clearly differentiated between the three single stimuli and the binary mixtures. Changes were observed in event-related potentials that reflected those obtained for intensity ratings in that they were observed for linalool and H2S in the linalool trained group only. The amplitude of the late 'endogenous' component (P3) was significantly decreased for these odors at frontal recording sites. In summary, strong and specific training effects were observed in intensity ratings for participants trained with the test odor (linalool), but not for those trained with a different odor. This was supported by a significant decrease of amplitudes of the event-related potentials at frontal recording sites following training with the test odor only     

Preparation of reminiscent aroma mixture of Japanese soy sauce

To prepare an aroma mixture of Japanese soy sauce by fewest components, the aroma concentrate of good sensory attributes was prepared by polyethylene membrane extraction, which could extract only the volatiles with diethyl ether. GC-MS-Olfactometry was done with the aroma concentrate, and 28 odor-active compounds were detected. Application of aroma extract dilution analysis to the separated fraction revealed high flavor dilution factors with respect to acetic acid, 4-hydroxy-2(or5)-ethyl-5(or2)-methyl-3(2H)-furanone (HEMF), 3-methyl-1-butanol (isoamyl alcohol), and 3-(methylsulfanyl)propanal (methional). A model aroma mixture containing above four odorants showed a good similarity with the aroma of the soy sauce itself. Consequently, the reminiscent aroma mixture of soy sauce was prepared in water. The ratio of acetic acid, HEMF, isoamyl alcohol, and methional was 2500:300:100:1.     

Concentration and Membrane Fluidity Dependence of Odor Discrimination in the Turtle Olfactory System

In the present study, we examined the concentration dependenceof odor discrimination in turtle olfactory bulbar responsesusing the cross-adaptation technique. In the odorant pairs withdiverse molecular structures, the degree of discrimination wasunchanged or only slightly decreased with an increase in odorantconcentrations, suggesting that odorants are well discriminatedeven at high concentrations. In the odorant pairs with closelyrelated molecular structures, the degree of discrimination wasdecreased with an increase in odorant concentrations. An increasein the temperature of turtle olfactory epithelium also decreasedthe ability to discriminate these odorants. There was a goodcorrelation between changes in the odor discriminating abilityinduced by an increase in odor concentrations and those inducedby a temperature increase. The liposomes were made of lipidsextracted from the turtle olfactory epithelia and changes oftheir membrane fluidity induced by adsorption of odorants weremonitored with DPH. There was a good correlation between a decreasein odor discriminating ability and the membrane fluidity changesinduced by odorants. We suggest that decreases in odor discriminatingability induced either by an increase in odor concentrationor by a temperature increase are ultimately caused by changesin the membrane fluidity. Chem. Senses 22: 553–563, 1997.     

Olfactory discrimination of aliphatic odorants at 1 ppm: too easy for CD-1 mice to show odor structure–activity relationships?

Using an operant conditioning paradigm we tested the ability of CD-1 mice to discriminate between 25 odorants comprising members of five homologous series of aliphatic odorants (C4-C8) presented at a gas phase concentration of 1 ppm. We found (a) that all mice significantly discriminated between all 50 stimulus pairs that involved odorants sharing the same functional group, but differing in carbon chain length, as well as between all 50 stimulus pairs that involved odorants sharing the same carbon chain length but differing in functional group, (b) a significant negative correlation between discrimination performance and structural similarity of odorants in terms of differences in carbon chain length with the acetic esters and the 2-ketones, but not with the 1-alcohols, n-aldehydes, and n-carboxylic acids tested, and (c) that odorant pairs differing in functional group were significantly better discriminated than odorant pairs differing in carbon chain length. These findings demonstrate that CD-1 mice have excellent discrimination ability for structurally related aliphatic odorants, that correlations between discrimination performance and structural similarity of odorants are odorant class-specific rather than a general phenomenon, and that both carbon chain length and type of functional group play an important role for odor quality coding in mice.     

Just noticeable differences in component concentrations modify the odor quality of a blending mixture

The odors we perceive are mainly the result of mixtures of odorants that, however, are commonly perceived as single undivided entities; nevertheless, the processes involved remain poorly explored. It has been recently reported that perceptual blending based on configural olfactory processing can cause odorant mixtures to give rise to an emergent odor not present in the components. The present study examined whether specific component proportions are required to elicit an emergent odor. Starting from the composition of a ternary target mixture in which an emergent pineapple odor was perceived, 4 concentration levels of each component were chosen to elicit just noticeable differences (JNDs). Each combination of levels was used to design sample mixtures. Fifteen subjects evaluated the intensity, typicality, and pleasantness of each sample mixture against the target mixture in a paired-comparison protocol. Statistical modeling showed that a variation of less than 1 JND in one of the components was sufficient to induce a significant decrease in pineapple odor typicality in the ternary mixture. This finding confirms previous findings on perceptual blending in simple odorant mixtures and underscores the human ability to discriminate between odor percepts induced by mixtures including very similar odorant proportions.     

Potent odorants characterize the aroma quality of leaves and stalks in raw and boiled celery

The raw and boiled odors of celery leaves and stalks were investigated. Among 12 compounds identified as potent odorants, 3-n-butylphthalide 1, sedanenolide 2, and trans- and cis-sedanolides 3, 4 were assessed to be most contributive to the overall odor of celery. These three phthalides, (3E,5Z)-1,3,5-undecatriene, myrcene, and (E)-2-nonenal were common to both raw and boiled materials. Two compounds, ((Z)-3-hexenal and (Z)-3-hexenol), were dominant in raw materials and four compounds, (2-methylbutanoic acid, sotolon, beta-damascenone, and beta-ionone), were dominant in boiled materials. Sensory evaluations were performed on natural celery odor and a series of reconstructed model aromas by assigning each intensity ratings for a set of seven odor qualities which aptly describe the odors of raw and boiled celery. According to the evaluation results, six common components contributed to the moderate odor of raw celery, two components dominant in raw materials enhanced the raw celery character, and four components dominant in boiled materials reduced the raw celery character and enhanced the boiled celery character. It was clarified that boiling-induced changes in celery odor were not affected by the amounts of phthalides, but by thermally generated compounds such as sotolon, beta-damascenone, and beta-ionone, which reduce the "green spicy" note.     

'Brain aided' musk design

This brief review, including new experimental results, is the summary of a talk at the RSC/SCI conference flavours & fragrances 2004 in Manchester, United Kingdom, 12-14 May, 2004. Musk odorants have been a classical domain for computer aided structure-odor relationship (SOR) studies, but, contrary to sandalwood or amber odorants, they belong to structurally very different substance classes, e.g., macrocycles, aromatic polycycles, and nitro arenes. Most SOR computer models are restricted to one class, excluding structural diversity to increase predictability. But even within a musk family, structural similarities are often due to a common synthetic access, and do not reflect binding requirements for the musk receptor. Beyond that, the importance of structural key features can be missed, which is discussed on the example of the (4S)-Me group of Galaxolide. By synthesis and olfactory evaluation of Galaxolide-like shaped macrobicycles as model compounds for conformationally constrained (12R)-12-methyltridecano-13-lactone, it was investigated how likely there is more than one musk receptor. Finally, the new family of so-called linear musks is discussed, especially with respect to the conformational importance of the gem-2',2'-dimethyl moiety in Helvetolide and the additional 2'-carbonyl group of Romandolide--structural features that strongly diminish the musk odor of macrocycles. On the example of 2-methyl-2-[(E)-1,2,4-trimethylpent-2-enyloxy]propyl esters, the 'brain-aided' design and conformational analysis of musk odorants is illustrated. The overview concludes with the synthesis, odor evaluation, and conformational discussion of the new musk odorant 2-(3,3-dimethylcyclohexyl)propanoic acid ethoxycarbonylmethyl ester.     

Olfactory perception in honeybees: Concatenated and mixed odorant stimuli,concentration, and exposure effects

Summary Here we present results obtained from 7 different series of experiments, all employing odor conditioning of proboscis extension in worker honeybees and each designed to address a particular question involving olfactory perception. The questions relate to: temporal complexity of odor cues; effects of concentration, suppression, and/or potentiation in mixture perception; acquisition and extinction rates, as well as levels of generalization associated with aliphatic compounds that have the same functional groups or same alkyl radical length; and the effects of continuous exposure to odorants in the first several days of adult life on various learning and discrimination tasks involving olfactory perception. From the data obtained in these experiments we were able to conclude the following: First, worker honeybees have a limited ability to perceive complex temporal odor-quality patterns in olfactory stimuli — they learn to associate the quality of only the last part of the stimulus with a sucrose reward. Second, we confirm that citral is qualitatively different in several perceptual contexts involving odor learning and conditioning and our results help elucidate the nature of these differences as they relate to learning, discrimination, mixture perception, and continuous exposure to particular odorants. Third, we appear to have uncovered some important perceptual differences between functional groups attached to the first as opposed to the second carbon atom of alkyl radicals. Finally, we failed to uncover any significant effects relating to continuous exposure to odorants during the first several days of a worker's adult life, despite evidence that considerable sensory development takes place during this period. Thus ontogenetic changes to the peripheral system due to environmental effects appear to leave basic perceptual systems unaltered.     

The perceptual similarity of substitued benzenes and pyridines as a funciton of steric hindrance

Differences between the odor percepts of 10 1,3-dialkylbenzenesand 10 2,6-dialkylpyridines are based mainly on two perceptualdimensions. The overall perceptual similarity of the identicallysubstituted benzene/pyridine pairs seems to depend on accessibilityto the pyridine nitrogen atom and steric hindrance to the aromaticring in a non-monotonic way. However, a linear relationshipbetween a unidimensional percept and steric hindrance exists.This conclusion is based on multidimensional scaling and odorprofiling studies. This set of odorants may help to elucidatethe receptor mechanisms underlying odor diserimination.     

Adaptation and cross-adaptation to odor stimulation of olfactory receptors in the tiger salamander

We have used the effects of self- and cross-adaptation on the unitary responses of olfactory receptors of the tiger salamander to odor stimulation to investigate the stimulus-specific components of these responses and to provide information about the cross-cell variations in the numbers and numbers of types of constitutent receptive sites. An olfactometer delivered sequential odorous pulses, either juxtaposed or separated by a variable time delay. We used four pairs of odorants judged to be similar within a given pair. The unitary response to the test stimulation relative to that of the conditioning stimulation varied from being unchanged to being completely eliminated. We sometimes observed substantial poststimulus increases in the firing rate following stimulation with juxtaposed odorous pulse. Except in the case of one odorant pair, cross-adaptation occurred both with juxtaposed pulses and with pulses separated in time. With the methyl butyrate/ethyl butyrate odorant pair, however, statistically significant cross-adaptation appeared only with juxtaposed pulses. We propose a simple model to aid in explaining these phenomena. The experimental observations in conjunction with this model are used to obtain estimates of the maximal and minimal number of receptive site types available for interaction with the chosen odorants.