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Behavior genetics of olfactory responses in Drosophila. I. Olfactometry and strain differences in Drosophila melanogaster.

A simple olfactometer was constructed for measuring the responses of adult Drosophila to odoriferous substances, and an index (attractability index, AI) is proposed to evaluate the response. Five strains of Drosophila melanogaster of different geographic origins were tested for their responses to five chemical attractants: ethyl alcohol, acetic acid, lactic acid, ethyl acetate, and n-butyraldehyde. Statistical analysis has shown that the differences of AI among strains are highly significant for all the odorants except acetic acid and that male flies are, in general, less sensitive than females. The number of flies which were inactive in the olfactometer also varied with strain, sex, and odorant, suggesting the involvement of some other behaviors, and this raises some difficulties in interpreting the heterogeneities of AI. Nevertheless, these results suggest that a considerable amount of genetic variation for olfactory response exists in natural population of D. melanogaster.

Acetates↗

Aroma compound analysis of Piper nigrum and Piper guineense essential oils from Cameroon using solid-phase microextraction-gas chromatography, solid-phase microextraction-gas chromatography-mass spectrometry and olfactometry.

The investigation of aroma compounds of the essential oils of dried fruits of black pepper (Piper nigrum) and black and white "Ashanti pepper" (Piper guineense) from Cameroon by means of solid-phase microextraction (SPME) was carried out for the first time to identify the odorous target components responsible for the characteristic odor of these valuable spices and food flavoring products. By means of GC-flame ionization detection (FID) and GC-MS (using different polar columns) the main compounds (concentration >3.0%, calculated as area of GC-FID analysis using a non-polar fused-silica open tubular RSL-200 column) of the SPME headspace samples of P. nigrum (black) and P. guineense (black and white) were found to be: P. nigrum (black)--germacrene D (11.01%), limonene (10.26%), beta-pinene (10.02%), alpha-phellandrene (8.56%), beta-caryophyllene (7.29%), alpha-pinene (6.40%) and cis-beta-ocimene (3.19%); P. guineense (black)--beta-caryophyllene (57.59%), beta-elemene (5.10%), bicyclogermacrene (5.05%) and alpha-humulene (4.86%); and P. guineense (white)--beta-caryophyllene (51.75%), cis-beta-ocimene (6.61%), limonene (5.88%), beta-pinene (4.56%), linalool (3.97%) and alpha-humulene (3.29%). The most intense odor impressions of the essential oils of the various dried pepper fruits were given byprofessional perfumers as follows: P nigrum (black)--fine, pleasant black pepper note; P. guineense (black)--black pepper top-note; and P. guineense (white)--pleasant white pepper note. These analytical results for the SPME headspace samples of three different pepper species from Cameroon are in accordance with the olfactoric data of the corresponding essential oils. A GC-sniffing technique was used to correlate the single odor impression of the identified SPME headspace volatiles of the three investigated pepper samples with the following results: themain compounds such as beta-caryophyllene, germacrene D, limonene, beta-pinene, alpha-phellandrene and alpha-humulene, as well as minor constituents such as delta-carene, beta-phellandrene, isoborneol, alpha-guaiene, sarisan, elemicin, calamenene, caryophyllene alcohol, isoelemicin, T-muurolol, cubenol and bulnesol, are of greatest importance for the characteristic pepper odor notes of these three Piper samples. Further aroma impressions can be attributed to mono- and sesquiterpenes, hexane, octane and nonane derivatives.

Chromatography, Gas↗

Identification of aroma active compounds in orange essence oil using gas chromatography-olfactometry and gas chromatography-mass spectrometry.

Using GC-MS and GC-flame ionization detection (FID)/olfactometry, 95 volatile components were detected in orange essence oil, of which 55 were aroma active. In terms of FID peak area the most abundant compounds were: limonene, 94.5%; myrcene, 1%; valencene, 0.8%; linalool, 0.7%, and octanal, decanal, and ethyl butyrate, 0.3% each. One hundred percent of the aroma activity was generated by slightly more than 4% of the total volatiles. The most intense aromas were produced by octanal, wine lactone, linalool, decanal, beta-ionone, citronellal, and beta-sinensal. Potent aroma components reported for the first time in orange essence oil include: E-2-octenal, 1-octen-3-ol, Z-4-decenal, E,E-2,4-nonadienal, guaiacol, gamma-octalactone, and m-cresol. Over 20 compounds were identified for the first time in orange essence oil using MS, however, most did not exhibit aroma activity.

Citrus↗

Electro-olfactogram (EOG) in olfactometry.

The electro-olfactogram (EOG) is a potential originated in olfactory epithelium evoked by odorous stimulation, which has been demonstrated and termed by Ottoson (1959). In order to ascertain the clinical application of EOG, we attempted to record similar potentials in human subjects. In some preliminary experiments using rabbits, it was quite easy to produce Ottoson's findings. In human subjects, however, there were sometimes great technical difficulties owing to the hidden anatomical position of the olfactory epithelium. Then we utilized Olympus selfoscope endoscopy for placing and retaining the electrode to the olfactory portion precisely and safely, with a tube for sending airborne odor. As results, we succeeded in recording negative spike discharge similar to Ottoson's findings. We examined a total of 27 nasal cavities in 22 patients with peripheral olfactory disorders and 11 nasal cavities in 12 patients with central olfactory disorders. The positive EOG rate in patients with peripheral or central olfactory disorders was 28 and 69%, respectively. Moreover, the positive rate was decreased in proportion to the degree of olfactory disorders in chronic rhinosinusitis. No positive EOG was observed in anosmic nasal cavities, while it was 68% in normal persons and resulted in an equal value to anosmia group caused by the damage of central olfactory pathway. From these results, it is considered that the positive EOG study is the only method which provides objective finding for the differential diagnosis of anosmia caused by disorders of olfactory epithelium or disorders of central olfactory tract.

Chronic Disease↗

Improved hardware and software for quick gas chromatography-olfactometry using CHARM and GC-"SNIF" analysis.

A multisniffing system has been developed to allow three panelists to simultaneously participate in a GC-olfactometric analysis. This device, associated with a computerized data treatment, allows shortening CHARM and GC-"SNIF' analyses to less than 1 week and less than 1 day, respectively. The program was developed as an extension of an existing commercial chromatography data system, as usual GC processing functions are suited to the treatment of olfactograms (plots of odor response versus GC elution time). Because of the improved algorithm, the consequences of gaps in coincident responses were minimized, and the systematic use of a panel improved the repeatability of CHARM olfactograms. Comparing both methods, GC-SNIF repeatability appears to be higher than that of CHARM, as the former method uses a larger panel, but in a shorter lapse of time.

Acyclic Monoterpenes↗

Identification and quantification of impact odorants of aged red wines from Rioja. GC-olfactometry, quantitative GC-MS, and odor evaluation of HPLC fractions.

An XAD-4 extract from a 5-year-old wine from Rioja (Spain) was analyzed by aroma extract dilution analysis. Most of the odorants were quantified by GC-MS. A second extract was fractionated in an HPLC system with a C-18 semipreparative column. Fifty fractions were recovered, their alcoholic degree and pH were further adjusted to those of the wine, and those fractions that showed strong odor characteristics were further re-extracted and analyzed by GC-O and GC-MS. Reconstitution experiments were carried out to confirm the role of the odorants detected in the fractions. Fifty-eight odorants were found in the Rioja wine, 52 of which could be identified. Methyl benzoate was found to be a wine aroma constituent for the first time. The most important odorants are 4-ethylguaiacol, (E)-whiskey lactone, 4-ethylphenol, beta-damascenone, fusel alcohols, isovaleric and hexanoic acids, eugenol, fatty acid ethyl esters, and ethyl esters of isoacids, Furaneol, phenylacetic acid, and (E)-2-hexenal. Comparison among the three techniques shows good agreement and demonstrates that they are complementary.

Chromatography, Gas↗

Use of GC-olfactometry to identify the hop aromatic compounds in beer.

This paper describes a sensorial aroma extract dilution analysis (AEDA) approach to the analysis of beer aromas derived from hops. To obtain an extract with an odor representative of the original product, the XAD extraction procedure was applied and the experimental conditions were optimized. The aromagrams of three beers were compared: one brewed without hops, one brewed with Saaz hop pellets, and one brewed with Challenger hop pellets. One spicy/hoppy compound, unmodified from hop to beer, proved responsible for the most intense odor in both hopped beer extracts. Another flavoring compound in hops, linalool, also survives through the process to the final beer. Other compounds such as gamma-nonalactone and humuladienone, although not found in our extracts of hop, significantly modify beer aromagrams after hopping. Sulfur compounds characteristic of Challenger hops proved to be at least partially responsible for the unpleasant flavor found in the corresponding beer.

Beer↗