Intranasal concentrations of orally administered flavors

The odorants emanating from the oral cavity during eating and drinking reach the olfactory mucosa via the pharynx (retronasal olfaction). It is unclear which variables influence the perception of intraorally applied substances. The aim of the present study was to determine the temporal profiles of volatile odor concentrations at different locations in the nasal cavity during consumption of liquid and solid custard samples using proton transfer reaction mass spectrometry. Intranasal odor concentrations were measured at least twice in nine subjects (six female, three male) at four nasal positions during the consumption of liquid and solid custards. The low-viscosity custard was swallowed earlier than the more solid one. The compounds were found to reach the nose in different concentrations. Largest maximal amplitudes were measured in the nasopharynx, whereas lowest concentrations were found in the region of the olfactory cleft. In addition, different odorants reached the different regions in the nasal cavity in varying concentrations, indicated by a significant interaction between factors "position" and "compound". Furthermore, the compounds were found to reach the positions within the nasal cavity with different latencies. These results indicate that different volatile flavor compounds exhibit different temporal and spatial profiles in terms of their intranasal distribution.     

Characterization of odorants in human urine using a combined chemo-analytical and human-sensory approach: a potential diagnostic strategy

The volatile and odorous profile of human urine may be a rich source for physiological information and could increase our understanding of metabolization and excretion processes of low-molecular weight compounds originating from, for example, dietary or endogenous sources. However, the diagnostic potential of the urinary volatile fraction is not yet fully understood, probably due to the limited application of modern analytical tools in urine volatile analysis. Accordingly, the aim of this study was to evaluate a combined chemo-analytical and human-sensory approach for characterization of the human urine odorant composition. We used one- and two-dimensional high resolution gas chromatography–olfactometry/mass spectrometry to identify commonly occurring and potent odorants in human urine. The studies were carried out on both native urine and on urine that had been treated by glucuronidase assays, with analysis of the liberated odor-active compounds using the same techniques. Based on retention indices, odor qualities and intensities, and mass spectra compared to references, a total of 14 odorants were detected in the majority of the untreated urine samples, and 24 odorants in the glucuronidase-treated samples. A major part of the identified substances are reported here for the first time. Our results show that chemosensory approaches are a useful strategy for the characterization of the odorant profile of human urine.     

Pheromone signalling in the mouse: role of urinary proteins and vomeronasal organ.

The lipocalin protein family is characterized in structure by a conserved hydrophobic pocket which can bind small volatile odorants. The Major Urinary Proteins (MUPs) are a class of lipocalins found in the urine of adult male mice which concentrate in the urine odorants which confer a characteristic odor. The behavioural as well as the endocrine effects of mouse urine and MUPs are briefly reviewed, suggesting a complex role is pheromonal communication. Some recent data on the molecular receptors of the vomeronasal organ further suggest a complex interaction with the MUP system.     

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.     

Urinary Volatile Compounds as Biomarkers for Lung Cancer: A Proof of Principle Study Using Odor Signatures in Mouse Models of Lung Cancer

A potential strategy for diagnosing lung cancer, the leading cause of cancer-related death, is to identify metabolic signatures (biomarkers) of the disease. Although data supports the hypothesis that volatile compounds can be detected in the breath of lung cancer patients by the sense of smell or through bioanalytical techniques, analysis of breath samples is cumbersome and technically challenging, thus limiting its applicability. The hypothesis explored here is that variations in small molecular weight volatile organic compounds (“odorants”) in urine could be used as biomarkers for lung cancer. To demonstrate the presence and chemical structures of volatile biomarkers, we studied mouse olfactory-guided behavior and metabolomics of volatile constituents of urine. Sensor mice could be trained to discriminate between odors of mice with and without experimental tumors demonstrating that volatile odorants are sufficient to identify tumor-bearing mice. Consistent with this result, chemical analyses of urinary volatiles demonstrated that the amounts of several compounds were dramatically different between tumor and control mice. Using principal component analysis and supervised machine-learning, we accurately discriminated between tumor and control groups, a result that was cross validated with novel test groups. Although there were shared differences between experimental and control animals in the two tumor models, we also found chemical differences between these models, demonstrating tumor-based specificity. The success of these studies provides a novel proof-of-principle demonstration of lung tumor diagnosis through urinary volatile odorants. This work should provide an impetus for similar searches for volatile diagnostic biomarkers in the urine of human lung cancer patients.     

The difference of urine between Brandt’s voleLasiopodomys brandtii in the breeding and non-breeding condition

Using animal house experiments we demonstrated that urine odor can elicit physiological changes in the maturation rate of Brandt’s vole (Radde, 1861). We showed that the urine of estrous females, anestrous females and non-breeding males had no effect on the time of sexual maturity of young voles. In contrast, the maturation rate of young female voles was accelerated, and the maturation rate of young male voles was slowed, by exposure to the urine of breeding adult males. Headspace analysis revealed significant differences in the levels of a subset of volatile constituents of urine from breeding and non-breeding males. From a total of 50 components, Propanoic acid, 2,5-Dimethylpyrazine, Dimethyldisulfide, 1-Octene, 1-Hexanol, Hexanoic acid and p-Xylene were observed only in the urine of breeding male Brandt’s voles. All other substances were present in the volatiles from urine of both breeding and non-breeding male Brandt’s vole.     

A simple and flexible device to odorize large stimulation areas

This paper describes a flow dilution olfactometer which allowsthe odorization of large stimulation areas and the easy manipulationof several odorants and/or concentrations. Generation of theodorized air is performed by mixing in two steps the odor vaporcontained in Tedlar bags with a pure air stream flowing continuouslyout of a nozzle. Discrete concentration values are obtainedby using pre-adjusted needle valves to change the vapor flowsampled in the bags. This kind of olfactometer was utilizedto study odor coding in the olfactory bulbs of rats and rabbits.Five Odorants were delivered at concentrations ranging from2 x 10^-4 to 1.5 x 10^-2 of the saturated vapor pressure. Measurementsshowed that lower concentrations can be obtained by fillingthe bags with a more diluted odor vapor. Furthermore, the numberof test odorants can be increased at low cost by increasingthe number of Tedlar bags.     

Perceptual blending in odor mixtures depends on the nature of odorants and human olfactory expertise

Our olfactory system is confronted with complex mixtures of odorants, often recognized as single entities due to odor blending (e.g., coffee). In contrast, we are also able to discriminate odors from complex mixtures (e.g., off-odors). Therefore, the olfactory system is able to engage either configural or elemental processes when confronted with mixtures. However, the rules that govern the involvement of these processes during odor perception remain poorly understood. In our first experiment, we examined whether simple odorant mixtures (binary/ternary) could elicit configural perception. Twenty untrained subjects were asked to evaluate the odor typicality of mixtures and their constituents. The results revealed a significant increase in odor typicality in some but not all mixtures as compared with the single components, which suggest that perceptual odor blending can occur only in specific mixtures (configural processing). In our second experiment, we tested the hypothesis that general olfactory expertise can improve elemental perception of mixtures. Thirty-two trained subjects evaluated the odor typicality of the stimuli presented during the first experiment, and their responses were compared with those obtained from the untrained panelists. The results support the idea that general training with odors increases the elemental perception of binary and ternary blending mixtures.     

Preliminary investigation of using volatile organic compounds from human expired air, blood and urine for locating entrapped people in earthquakes

A preliminary investigation on the possibility of using volatile organic compounds (VOCs) determination of expired air, blood and urine, for the early location of entrapped people in earthquakes, has been carried out. A group of 15 healthy subjects has been sampled. The identification of a common "core" of substances might provide indications of human presence that can be used for the development of a real time field analytical method for the on site detection of entrapped people. Expired air samples have been analyzed by thermal desorption GC/MS and VOCs from blood and urine by headspace SPME-GC/MS. Acetone was the only compound found common in all three matrices. Isoprene was found in both expired air and blood samples. Acetone and isoprene along with a number of saturated hydrocarbons were among the major constituents identified in expired air analysis. Various ketones (2-pentanone, 4-heptanone, 2-butanone) were also determined over urine specimens. Using the techniques and methods of field analytical chemistry and technology appears to be the proper approach for applying the results of the present study in real situations.     

Origin, functions and chemistry of H‐2 regulated odorants

Genes located within the major histocompatibility complex (MHC) of mice are responsible for individual differences in body odor (odortypes). In this review we suggest that the MHC genes themselves are responsible for odor differences among MHC‐congenic mice. Recent studies indicating that volatile carboxylic acids are at least in part responsible for the individual odors and what this finding implies about the pathway from gene to odorant are also reviewed. We suggest that odorants or their precursors are bound directly by MHC products and are released into serum and concentrated in urine. Finally, possible functions of MHC odortypes in mice are enumerated and important future research questions are raised. This revised version was published online in July 2006 with corrections to the Cover Date.     

Canonical Discriminant Analysis Applied to the Headspace GC Profiles of Coffee Cultivars

Chemometric methods were applied for analyzing the relationship between the classification of coffee cultivars and their volatile components. Six typical cultivars were selected from Coffea arabica L., and their headspace profiles were analyzed by GC and GC/MS. A canonical discriminant analysis, with GC peaks as the variables, suggested the existence of a relationship between the sensory characteristics and canonical variables.

The six coffee cultivars were divided into three groups, respectively having a roasted note, sweet aroma and fermented odor. It is suggested that Brazil and Mandheling varieties had a roasted note derived from methylpyrazine, while Mocha coffee had a fermented odor derived from 2,3-butanedione.     

Olfactory discrimination ability of human subjects for ten pairs of enantiomers.

We tested the ability of human subjects to distinguish between enantiomers, i.e. odorants which are identical except for chirality. In a forced-choice triangular test procedure 20 subjects were repeatedly presented with 10 enantiomeric odor pairs and asked to identify the bottle containing the odd stimulus. We found (i) that as a group, the subjects were only able to significantly discriminate the optical isomers of alpha-pinene, carvone and limonene, whereas they failed to distinguish between the (+)- and (-)-forms of menthol, fenchone, rose oxide, camphor, alpha-terpineol, beta-citronellol and 2-butanol; (ii) marked individual differences in discrimination performance, ranging from subjects who were able to significantly discriminate between 6 of the 10 odor pairs to subjects who failed to do so with 9 of the 10 tasks; (iii) that with none of the 10 odor pairs were the antipodes reported to differ significantly in subjective intensity when presented at equal concentrations; and (iv) that error rates were quite stable and did not differ significantly between sessions, and thus, we observed a lack of learning or training effects. Additional tests of the degree of trigeminality and threshold measurements of the optical isomers of alpha-pinene, carvone and limonene suggest that the discriminability of these three enantiomeric odor pairs is indeed due to differences in odor quality. These findings support the assumption that enantioselective molecular odor receptors may only exist for some but not all volatile enantiomers and thus that chiral recognition of odorants may not be a general phenomenon but is restricted to some substances.     

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.     

Phosphoinositide 3-Kinase Dependent Inhibition as a Broad Basis for Opponent Coding in Mammalian Olfactory Receptor Neurons

Phosphoinositide 3-kinase (PI3K) signaling has been implicated in mediating inhibitory odorant input to mammalian olfactory receptor neurons (ORNs). To better understand the breadth of such inhibition in odor coding, we screened a panel of odorants representing different chemical classes, as well as odorants known to occur in a natural odor object (tomato), for their ability to rapidly activate PI3K-dependent inhibitory signaling. Odorants were screened on dissociated native rat ORNs before and after pre-incubation with the PI3K-isoform specific blockers AS252424 and TGX221. Many different odorants increased their excitatory strength for particular ORNs following PI3K blockade in a manner consistent with activating PI3K-dependent inhibitory signaling in those cells. The PI3K-dependent inhibitory odorants overlapped with conventional excitatory odorants, but did not share the same bias, indicating partial partitioning of the odor space. Finding that PI3K-dependent inhibition can be activated by a wide range of otherwise conventional excitatory odorants strongly implies PI3K-dependent inhibition provides a broad basis for opponent coding in mammalian ORNs.     

Identification of the possible new odor-active compounds “12-methyltridecanal and its analogs” responsible for the characteristic aroma of ripe Gouda-type cheese

The aroma concentrates of the three maturation stages of Gouda-type cheeses were prepared by combining the solvent extraction and the solvent assisted flavor evaporation techniques. The aroma extract dilution analysis applied to the volatile fraction revealed 31 odorants that were identified or tentatively identified from the 38 odor-active peaks with FD factors between 4³ and 4⁸. By comparison with the FD factors in the three maturation stages of the cheeses, 16 odorants, including 12-methyltridecanal, which is a newly identified odorant from the cheese, increased with the increasing maturation stage of the cheese. In addition, many iso- and anteiso-methyl-branched long-chain aliphatic aldehydes could be identified as the analogs of 12-methyltridecanal, which have a unique odor note. It may be then expected that these aldehydes were able to influence the flavor of the highly ripened Gouda cheese, since these compounds also increased with the increasing maturation stage.     

Olfactory discrimination ability and odor structure-activity relationships in honeybees.

Using the training procedure introduced by von Frisch in 1919, we tested the ability of free-flying honeybees to discriminate a conditioning odor from an array of 44 simultaneously presented substances. The stimuli included homologous series of aliphatic alcohols, aldehydes and ketones, isomeric forms of some of these substances, as well as several terpenes and odor mixtures, and thus comprised stimuli of varying degrees of structural similarity to any conditioning odor. We found (i) that the honeybees significantly distinguished between 97.0% of the 1848 odor pairs tested, thus showing an excellent discrimination performance when tested in a free-flying situation with an array of structurally related substances; (ii) a significant negative correlation between discrimination performance and structural similarity of odorants in terms of differences in carbon chain length with all aliphatic substance classes tested; (iii) that both the position and type of a functional group also affected discriminability of odorants in a substance class-specific manner; and (iv) striking similarities in odor structure-activity relationships between honeybees and human and nonhuman primates tested previously on a subset of substances employed here. Our findings demonstrate that the similarities found in the structural organization of the olfactory systems of insects and vertebrates are paralleled by striking similarities in relative discrimination abilities. This strongly suggests that similar mechanisms of odor coding and discrimination may underlie olfaction in vertebrates and insects.     

Olfactory discrimination ability for aromatic odorants as a function of oxygen moiety.

To assess the significance of the type of oxygen moiety on odor quality of aromatic compounds, I tested the ability of human subjects to distinguish between odorants sharing a benzene ring and the same total number of carbon atoms but differing in their functional groups. Phenyl ethanol, phenyl acetaldehyde, phenyl methyl ketone, methyl benzoate and phenyl acetic acid, were employed. In a forced-choice triangular test procedure 20 subjects were repeatedly presented with all possible binary combinations of the five odorants, and asked to identify the bottle containing the odd stimulus. I found (i) that as a group, the subjects performed significantly above chance level in six of the tasks whereas they failed to do so with the four other tasks; (ii) marked interindividual differences in discrimination performance, ranging from subjects who were able to significantly distinguish between all 10 odor pairs to subjects who failed to do so with the majority of the tasks; and (iii) that odor pairs that involved methyl benzoate or phenyl methyl ketone were significantly easier to discriminate than those that involved phenyl acetaldehyde or phenyl ethanol, and thus there was a clear dependence of discriminability on type of functional group. Additional tests of the degree of trigeminality of the five aromatic substances indicated that the discriminability of the odor pairs is indeed due to differences in odor quality. A comparison of the present results with those of an earlier study that employed aliphatic odorants suggests that functional oxygen-containing groups may generally be an important determinant of the interaction between the stimulus molecule and the olfactory receptor, and thus may generally be a molecular property affecting odor quality in a substance class-specific manner. The poorer discriminatory performance of the subjects with aromatic odorants compared with corresponding aliphatic substances suggests that the structure of the alkyl rest attached to a functional group may also play a crucial role for recognition of ligands at the olfactory receptor and thus for odor quality.     

Combinatorial receptor codes for odors

The discriminatory capacity of the mammalian olfactory system is such that thousands of volatile chemicals are perceived as having distinct odors. Here we used a combination of calcium imaging and single-cell RT-PCR to identify odorant receptors (ORs) for odorants with related structures but varied odors. We found that one OR recognizes multiple odorants and that one odorant is recognized by multiple ORs, but that different odorants are recognized by different combinations of ORs. Thus, the olfactory system uses a combinatorial receptor coding scheme to encode odor identities. Our studies also indicate that slight alterations in an odorant, or a change in its concentration, can change its "code," potentially explaining how such changes can alter perceived odor quality.     

Retention of components in a mixture of volatile organic substances by maltodextrins

The effect of composition and origin of maltodextrins on the retention of 38 components in a mixture of volatile organic substances (odorants) during 6-month storage was studied by means of capillary gas-liquid chromatography. The retention of esters increased with an increase in their molecular weight. The retention of lactones, phenols, linalool, menthone, and damascone was 75-85%. Storage of aldehydes was accompanied by oxidation, and the retention of these substances did not exceed 55%. The retention of odorants increased with a decrease in the molecular weight of maltodextrins. The maximum retention was typical of maltodextrin from amylopectin starch not containing amylose.     

Evaluation of the key odorants of foods by dilution experiments, aroma models and omission.

The state of the art in aroma analysis is reviewed with emphasis on aroma-recombination studies using synthetic blends of odorants (aroma models) which have been prepared on the basis of analytical data. The model that matches the original aroma is the starting material for omission experiments which are performed to establish the odorants that actually contribute to the aroma. These experiments are discussed in detail for the aromas of two wine varieties, three olive oils of different provenance, French fries, boiled beef and coffee. The results indicate that odorants with higher odour activity values (OAV, the ratio of the concentration to the odour threshold) are frequently essential for the aroma. However, there are exceptions where odorants with high OAVs are suppressed in the aroma and compounds with lower OAVs are important contributors. These findings are discussed in the light of model experiments which have been reported in the literature to obtain an insight into the perceptual interactions of odorants in mixtures.