Geosmin, a Food- and Water-Deteriorating Sesquiterpenoid and Ambivalent Semiochemical, Activates Evolutionary Conserved Receptor OR11A1.

Geosmin, a ubiquitous volatile sesquiterpenoid of microbiological origin, is causative for deteriorating the quality of many foods, beverages, and drinking water, by eliciting an undesirable "earthy/musty" off-flavor. Moreover, and across species from worm to human, geosmin is a volatile, chemosensory trigger of both avoidance and attraction behaviors, suggesting its role as semiochemical. Volatiles typically are detected by chemosensory receptors of the nose, which have evolved to best detect ecologically relevant food-related odorants and semiochemicals. An insect receptor for geosmin was recently identified in flies. A human geosmin-selective receptor, however, has been elusive. Here, we report on the identification and characterization of a human odorant receptor for geosmin, with its function being conserved in orthologs across six mammalian species. Notably, the receptor from the desert-dwelling kangaroo rat showed a more than 100-fold higher sensitivity compared to its human ortholog and detected geosmin at low nmol/L concentrations in extracts from geosmin-producing actinomycetes.

Volatile Profiling of Spirulina Food Supplements.

Spirulina, a cyanobacterium widely used as a food supplement due to its high nutrient value, contains volatile organic compounds (VOCs). It is crucial to assess the presence of VOCs in commercial spirulina products, as they could influence sensory quality, various processes, and technological aspects. In this study, the volatile profiles of seventeen commercial spirulina food supplements were determined using headspace solid-phase microextraction (HS-SPME), coupled with gas chromatography-mass spectrometry (GC-MS). The identification of volatile compounds was achieved using a workflow that combined data processing with software tools and reference databases, as well as retention indices (RI) and elution order data. A total of 128 VOCs were identified as belonging to chemical groups of alkanes (47.2%), ketones (25.7%), aldehydes (10.9%), alcohols (8.4%), furans (3.7%), alkenes (1.8%), esters (1.1%), pyrazines (0.8%), and other compounds (0.4%). Major volatiles among all samples were hydrocarbons, especially heptadecane and heptadec-8-ene, followed by ketones (i.e., 4-(2,6,6-trimethyl-1-cyclohexen-1-yl)-3-buten-2-one, ß-ionone, 2,2,6-trimethylcyclohexan-1-one), aldehydes (i.e., hexanal), and the alcohol oct-1-en-3-ol. Several volatiles were found in spirulina dietary supplements for the first time, including 6,10-dimethylundeca-5,9-dien-2-one (geranylacetone), 6,10,14-trimethylpentadecan-2-one, hept-2-enal, octanal, nonanal, oct-2-en-1-ol, heptan-1-ol, nonan-1-ol, tetradec-9-en-1-ol, 4,4-dimethylcyclohex-2-en-1-ol, 2,6-diethylpyrazine, and 1-(2,5-dimethylfuran-3-yl) ethanone. The methodology used for VOC analysis ensured high accuracy, reliability, and confidence in compound identification. Results reveal a wide variety of volatiles in commercial spirulina products, with numerous newly discovered compounds, prompting further research on sensory quality and production methods.

Key Odorants Forming Aroma of Polish Mead: Influence of the Raw Material and Manufacturing Processes.

Mead was analyzed by using the concept of molecular sensory science for the identification of key odorants. A total of 29 odor-active compounds were identified in mead by using gas chromatography olfactometry (GCO). Flavor dilution (FD) factors of identified compounds ranged from 1 to 16,384, compounds with FD factors ≥32 were quantitated by using stable isotopically substituted odorants as internal standards or external standard method, and odor activity values (OAVs) were calculated. Fifteen compounds showed OAVs ≥1: aldehydes (2-phenylacetaldehyde, 3-(methylsulfanyl)propanal), 4-hydroxy-3-methoxybenzaldehyde), esters (ethyl 3-methylbutanoate, ethyl propanoate, ethyl octanoate), alcohols (2-phenylethan-1-ol, 3- and 2-methylbutan-1-ol, 3-(methylsulyfanyl)propan-1-ol), furanons (4-hydroxy-2,5-dimethylfuran-3(2H)-one, 3-hydroxy-4,5-dimethylfuran-2(5H)-one), acids (3- and 2-methylbutanoic acid, acetic acid), 1,1-diethoxyethane, and 4-methylphenol. 2-Phenylacetaldehyde (OAV, 3100) was suggested as the compound with the biggest influence on the aroma of mead, followed by 4-hydroxy-2,5-dimethylfuran-3(2H)-one (OAV, 1900), 3-(methylsulfanyl)propanal (OAV, 890), and 2-phenylethan-1-ol (OAV, 680). Quantitative olfactory profile analysis revealed strong honey, malty, and alcoholic impressions. Omission experiments revealed that 3-(methylsulfanyl)propanal, 2-phenylethan-1-ol, 4-hydroxy-2,5-dimethylfuran-3(2H)-one, ethyl propanoate, ethyl 3-methylbutanoate, 2-phenylacetaldehyde, 3- and 2-methylbutanoic acid, 3-hydroxy-4,5-dimethylfuran-2(5H)-one, and 4-hydroxy-3-methoxybenzaldehyde were the key odorants in the mead. Determining concentrations of key odorants in important production steps showed that the fermentation and maturation stages had the strongest effect on the formation of mead aroma.

Identification of Bis(methylsulfanyl)methane and Furan-2(5H)-one as Volatile Marker Compounds for the Differentiation of the White Truffle Species Tuber magnatum and Tuber borchii.

Some truffles are expensive and, therefore, are prone to food fraud. A particular problem is the differentiation of high-priced Tuber magnatum truffles from cheaper Tuber borchii truffles, both of which are white truffles with similar morphological characteristics. Using an untargeted approach, the volatiles isolated from samples of both species were screened for potential marker compounds by comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC×GC-TOFMS) and statistical analysis of the obtained semiquantitative data. Results suggested bis(methylsulfanyl)methane and furan-2(5H)-one as compounds characterizing T. magnatum and T. borchii, respectively. Exact quantitation of both volatiles by conventional one-dimensional gas chromatography-mass spectrometry in combination with stable isotopologues of the target compounds as internal standards confirmed both as marker compounds. The method is suitable to be used in the routine analysis for the objective species differentiation of T. magnatum and T. borchii.

Molecular Background of the Lychee Aroma of Vitis vinifera L. 'Muscaris'.

Muscaris is a modern white grape variety with good fungal resistance and a pleasant aroma, the molecular background of which was unknown. A comparative aroma extract dilution analysis applied to Muscaris grapes and grapes of the father variety Muskateller revealed little differences and resulted in 39 and 35 odorants, respectively. Sixteen odorants exceeded their odor threshold concentrations. Odor reconstitution and omission experiments showed that the distinct lychee note in the aroma of the Muscaris grapes was generated by the combination of (2S,4R)-rose oxide and geraniol. This finding will guide further molecular research on the transfer of the lychee note into wine and may also be helpful for the targeted breeding of new grape varieties.

Peracetic acid residues in orange juice can lead to a 5-vinylguaiacol-induced clove-like off-flavor via Baeyer-Villiger oxidation of hesperidin.

A balanced flavor is a major quality attribute of orange juice. Formation of 4-vinylguaiacol during storage can lead to an undesirable clove-like off-flavor. However, clove-like off-flavors were occasionally reported despite low 4-vinylguaiacol concentrations, suggesting an alternative molecular background. Application of gas chromatography-olfactometry and aroma extract dilution analysis to an orange juice with a pronounced clove-like off-flavor resulted in the identification of 5-vinylguaiacol. The compound showed the same odor as 4-vinylguaiacol, but was previously unknown in orange juice. In five of six commercial orange juices with clove-like off-flavors, 5-vinylguaiacol was even more odor-active than 4-vinylguaiacol. Spiking and model studies suggested that 5-vinylguaiacol is formed during pasteurization from the natural orange juice component hesperidin and residual peracetic acid used as cleaning agent by a Baeyer-Villiger oxidation. An activity-guided screening approach confirmed the role of hesperidin as 5-vinylguaiacol precursor. In conclusion, peracetic acid should no longer be used in orange juice processing plants.

Sotolon and (2E,4E,6Z)-Nona-2,4,6-trienal Are the Key Compounds in the Aroma of Walnuts.

Fresh kernels of the walnut tree (Juglans regia L.) show a characteristic and pleasant aroma, the molecular basis of which was unknown. The application of an aroma extract dilution analysis resulted in 50 odor-active compounds. Among them, 37 had not been reported as fresh walnut kernel volatiles before, including the two odorants with the highest flavor dilution factors, namely, fenugreek-like smelling 3-hydroxy-4,5-dimethylfuran-2(5H)-one (sotolon) and oatmeal-like smelling (2E,4E,6Z)-nona-2,4,6-trienal. Quantitations revealed 17 odorants with concentrations in the walnuts that exceeded their odor threshold concentrations. Aroma reconstitution and omission experiments finally showed that the characteristic aroma of fresh walnuts is best represented by a binary mixture of sotolon and (2E,4E,6Z)-nona-2,4,6-trienal. Of both, the natural concentration was ∼10 µg/kg. Further sensory studies showed that the walnut character is intensified when their concentrations are in parallel increased to ∼100 µg/kg. This finding may guide the future breeding of new walnut cultivars with improved aroma.

Molecular Rearrangement of Four Typical Grape Free Terpenes in the Wine Environment.

In order to analyze the molecular rearrangement of terpenes in wine during aging, the changes in linalool, α-terpineol, nerol, and geraniol in model wine were investigated in the dark at low temperature for 90 days. Headspace-gas chromatograph-mass spectrometer/olfactometry was used for qualitative and relative quantitation of terpenes. Quantum mechanical calculation was used to analyze the Gibbs free energy. The results showed that nerol was converted into d-limonene, terpinolene, linalool, and α-terpineol. Geraniol was converted into ß-ocimene, terpinolene, and linalool. Linalool was converted into terpinolene. The conversion rate of nerol to terpinolene was the highest with 5.94%. α-Terpineol was not converted spontaneously into other terpenes due to its lowest Gibbs free energy, indicating that the cyclization and isomerization could occur spontaneously through an exotherm reaction. However, the dehydroxylation of linalool, nerol, and geraniol required an energy source.

Decoding the Fine Flavor Properties of Dark Chocolates.

Fine flavor properties of chocolates such as fruity, floral, and cocoa-like were decoded on a molecular level for the first time. The molecular compositions of six chocolates made out of liquors that were referenced with specific sensory attributes were analyzed. After the screening for odor-active molecules by aroma extract dilution analysis, selected compounds were quantitated with the overall aim to decode the distinct fine flavor attributes on a molecular level. Acidic and fruity flavor notes were associated with high dose over threshold factors (DoT factors) of acetic acid and fruity smelling esters such as ethyl 2-methylbutanaote, ethyl 3-methylbutanoate, and 3-methylbutyl acetate, respectively. Cocoa-like and roasty flavor notes were associated with high DoT factors for 2-methylbutanal, 3-methylbutanal, 4-hydroxy-2,5-dimethylfuran-3(2H)-one, and dimethyltrisulfane. The floral and astringent flavors were linked to high DoT factors of (-)-epicatechin, procyanidin B2, procyanidin C1, and 2-phenylethan-1-ol.

Influence of Varying Fermentation Parameters of the Yeast Strain Cyberlindnera saturnus on the Concentrations of Selected Flavor Components in Non-Alcoholic Beer Focusing on (E)-ß-Damascenone.

The diversification of beer flavor is becoming increasingly popular, especially in the field of non-alcoholic beers, where sales are growing steadily. While flavor substances of traditional beers can largely be traced back to defined secondary metabolites, the production of non-alcoholic beers with non-Saccharomyces yeasts generates novel fruity flavors, some of which cannot yet be assigned to specific flavor substances. In a recently published study, besides pear, cool mint sweets, and banana-like flavor, distinctive red berry and apple flavors were perceived in a non-alcoholic beer fermented with the yeast strain Cyberlindnera saturnus TUM 247, whose secondary metabolites were to be elucidated in this study. The trials were carried out using response surface methodology to examine the fermentation properties of the yeast strain and to optimize the beer with maximum fruitiness but minimal off-flavors and ethanol content. It turned out that a low pitching rate, a moderate fermentation temperature, and an original gravity of 10.5 °P gave the optimal parameters. Qualitative analysis of the secondary metabolites, in addition to standard analysis for traditional beers, was first performed using headspace-gas chromatography with olfactometry. (E)-ß-damascenone emerged as the decisive substance for the red berry and apple flavor and so this substance was then quantitated. Although (E)-ß-damascenone is a well-known secondary metabolite in beer and this substance is associated with apple or cooked apple- and berry-like flavors, it has not yet been reported as a main flavor component in non-alcoholic beers.

Impact of Malt Extract Addition on Odorants in Wheat Bread Crust and Crumb.

Application of gas chromatography-olfactometry and aroma extract dilution analysis to the volatiles isolated from (1) crust and (2) crumb of a wheat bread made with the addition of a dark liquid malt extract (LME) to the dough and (3) crust and (4) crumb of a reference bread made without addition resulted in the identification of 23 major odorants. Their quantitation followed by the calculation of odor activity values (OAV = ratio of concentration to odor threshold value) suggested that LME addition influenced the aroma of the bread predominantly by increasing seasoning-like smelling sotolon in crust and crumb, and caramel-like smelling compounds maltol and 4-hydroxy-2,5-dimethylfuran-3(2H)-one (HDMF) in the crumb. The increase in sotolon and maltol was explainable by direct transfer from the LME to the bread, whereas HDMF must have been formed from LME-derived precursors. This difference needs to be considered in the targeted optimization of LMEs for bread making.

Key Odorants of Raw and Cooked Green Kohlrabi (Brassica oleracea var. gongylodes L.).

Volatile compounds of raw and cooked green kohlrabi were investigated using a sensomics approach. A total of 55 odor-active compounds were detected and identified in raw and cooked green kohlrabi using GC-O. Twenty-eight odor-active compounds with high flavor dilution (FD) factors ranging from 64 to 1024 were quantitated, and odor activity values (OAVs) were determined. Eight compounds showed high OAVs in raw and cooked kohlrabi: five sulfur compounds (dimethyl trisulfide, methyl 2-methyl-3-furyl disulfide, and three isothiocyanates (1-isothiocyanato-3-(methylsulfanyl)propane, benzyl isothiocyanate, and 1-isothiocyanato-4-(methylsulfanyl)butane)), two lipid oxidation products (1-octen-3-one and trans-4,5-epoxy-(2E)-dec-2-enal), and 2-isopropyl-3-methoxypyrazine. Among these, the sulfur compounds contributed most to the overall smell of the raw and cooked vegetables. The quantitation analysis indicates that the eight odorants are the backbone compounds for raw and cooked kohlrabi. The OAVs for the backbone compounds and also for minor odorants are clearly higher in raw kohlrabi than in the cooked one. Differences can be explained by the influence of the cooking process.

Molecular Insights into the Contribution of Specialty Barley Malts to the Aroma of Bottom-Fermented Lager Beers.

Specialty barley malts provide unique aroma characteristics to beer; however, the transfer of specialty malt odorants to beer has not yet been systematically studied. Therefore, three beers were brewed: (1) exclusively with kilned base barley malt, (2) with the addition of a caramel barley malt, and (3) with the addition of a roasted barley malt. Major odorants in the beers were identified by aroma extract dilution analysis followed by quantitation and calculation of odor activity values (OAVs). The caramel malt beer was characterized by outstandingly high OAVs for odorants such as (E)-ß-damascenone, 2-acetyl-1-pyrroline, methionol, 2-ethyl-3,5-dimethylpyrazine, and 4-hydroxy-2,5-dimethylfuran-3(2H)-one, whereas the highest OAV for 2-methoxyphenol was obtained in the roasted malt beer. Quantifying odorants in the malts revealed that the direct transfer from malt to beer played only a minor role in the amount of malt odorants in the beers, suggesting a substantial formation from precursors and/or a release of encapsulated odorants during brewing.

Impact of Water on Odor-Active Compounds in Fermented and Dried Cocoa Beans and Chocolates Made thereof.

The impact of water on odor-active compounds in fermented and dried cocoa beans as well as in chocolate either produced by a novel processing (NPC) or a traditional processing (TPC) technology from the same batch of cocoa beans was investigated in this study. Quantitation of selected key odorants revealed significantly higher concentrations of Strecker aldehydes such as 3-(methylsulfanyl)propanal (66-fold) and phenylacetaldehyde (50-fold) after water treatment of the cocoa beans. The comparison of the two chocolates showed that higher amounts of the Strecker aldehydes 2-methylbutanal, 3-methylbutanal, and phenylacetaldehyde are released with water in the NPC (24-fold to 39-fold), compared to the TPC (7.3-fold-11-fold). In addition to Strecker aldehydes, the concentrations of many further characteristic key odorants of cocoa and chocolate increased after water treatment. Based on the results, a more intense retronasal odor perception of the analyzed compounds is expected due to their release during consumption in contact with saliva.

Molecular Background of a Moldy-Musty Off-Flavor in Cocoa.

The parallel application of aroma extract dilution analysis to the volatiles isolated from a sample of fermented cocoa seeds with a pronounced moldy-musty off-flavor and to the volatiles isolated from a flawless reference sample revealed (-)-geosmin, 4-methoxy-2,5-dimethylfuran-3(2H)-one, 1H-indole, and 3-methyl-1H-indole as potential off-flavor compounds on the basis of their odor quality and higher flavor dilution factors in off-flavor cocoa than in the reference sample. Quantitation of the four compounds in nine off-flavor cocoa samples and calculation of odor activity values (ratio of the concentrations to the odor threshold values) suggested the crucial roles of (-)-geosmin and 3-methyl-1H-indole for the off-flavor. In the chocolate industry, their quantitation can be used to objectively assess the off-flavor at the level of incoming goods inspection. Because both compounds are inhomogeneously distributed between the testa and the embryo, separate quantitation in the two parts of the seeds is required.

Molecular background of the undesired odor of polypropylene materials and insights into the sources of key odorants.

Screening the volatiles isolated from a standard polypropylene material consisting of a polypropylene homopolymer, the filler talcum, and a mixture of antioxidants, for odor-active compounds by application of an aroma extract dilution analysis revealed 30 odorants with flavor dilution factors ranging from 1 to 64. Eighteen odor-active compounds were subsequently quantitated by gas chromatography-mass spectrometry using stable isotopically substituted odorants as internal standards, and their odor activity values (OAVs) were calculated as ratios of the concentrations to the odor threshold values. Five odorants showed OAVs ≥1, among which were hex-1-en-3-one (OAV 12), butanoic acid (OAV 3), as well as 4-methylphenol, butan-1-ol, and 2-tert-butylphenol (all OAV 1). A comparative analysis of polypropylene materials with different additives suggested plastic-like, pungent smelling hex-1-en-3-one as an ubiquitous key odorant. Odor-active amounts of alkylphenols, in particular plastic-like, phenolic smelling 2-tert-butylphenol, were additionally formed in the presence of talcum and phenolic antioxidants. Whereas the precursors of the phenols were thus obvious, the origin of hex-1-en-3-one was unknown. Injection molding showed only little influence on odorant concentrations.

Characterization of Odorants Causing Smoky Off-Flavors in Cocoa.

Application of an aroma extract dilution analysis in parallel applied to the volatiles isolated from a sample of fermented cocoa with a pronounced smoky off-flavor and a reference sample with flawless aroma revealed seven potential off-flavor compounds with smoky/phenolic odor qualities and higher flavor dilution factors in the off-flavor sample than in the reference sample. These compounds were 2-methoxyphenol, 3- and 4-methylphenol, 3- and 4-ethylphenol, and 3- and 4-propylphenol. Their quantitation in seven off-flavor samples and two reference samples without off-flavor showed that 4-propylphenol did not exceed its odor threshold value. From the concentrations obtained for the other six compounds and their odor threshold values in deodorized cocoa butter, maximum tolerable concentrations were derived for the incoming goods inspection in the chocolate industry. The suggested limits are 2 µg/kg for 3-ethylphenol and 3-propylphenol, 20 µg/kg for 4-methylphenol, 3-methylphenol, and 4-ethylphenol, and 70 µg/kg for 2-methoxyphenol.

Enantioselective synthesis of tri-deuterated (-)-geosmin to be used as internal standard in quantitation assays.

For the accurate and sensitive quantitation of the off-flavor compound geosmin, particularly in complex matrices, a stable isotopologue as internal standard is highly advantageous. In this work, we present a versatile synthetic strategy leading from (4aR)-1,4a-dimethyl-4,4a,5,6,7,8-hexahydronaphthalen-2(3H)-one to tri-deuterated (-)-geosmin ((4S,4aS,8aR)-4,8a-dimethyl(3,3,4-2 H3 )octahydronaphthalen-4a(2H)-ol). The starting material was readily accessible from inexpensive 2-methylcyclohexan-1-one using previously published procedures.

Characterization of Odor-Active Compounds, Polyphenols, and Fatty Acids in Coffee Silverskin.

For the first time the volatile fraction of coffee silverskin has been studied focusing on odor-active compounds detected by gas chromatography-olfactometry/flame ionization detector (GC-O/FID) system. Two approaches, namely headspace (HS) analysis by solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) and odor-active compounds analysis by gas chromatography-olfactometry/flame ionization detector (GC-O/FID), have been employed to fully characterize the aroma profile of this by-product. This work also provided an entire characterization of the bioactive compounds present in coffee silverskin, including alkaloids, chlorogenic acids, phenolic acids, flavonoids, and secoiridoids, by using different extraction procedures and high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) system. Coffee silverskin was shown to be a good source of caffeine and chlorogenic acids but also of phenolic acids and flavonoids. In addition, the fatty acid composition of the coffee silverskin was established by GC-FID system. The results from this research could contribute to the development of innovative applications and reuses of coffee silverskin, an interesting resource with a high potential to be tapped by the food and nutraceutical sector, and possibly also in the cosmetics and perfumery.

Key Odorants in Japanese Roasted Barley Tea (Mugi-Cha)-Differences between Roasted Barley Tea Prepared from Naked Barley and Roasted Barley Tea Prepared from Hulled Barley.

The volatiles isolated by solvent extraction and solvent-assisted flavor evaporation (SAFE) from roasted barley tea, prepared from either hulled barley or naked barley, were subjected to a comparative aroma extract dilution analysis, which resulted in 27 odor-active compounds with flavor dilution factors (FD factors) of 64-1024. An additional 5 odorants were detected by static headspace analysis. Quantitation of these 32 compounds revealed 22 and 23 odorants in the naked barley tea and in the hulled barley tea, respectively, that exceeded their odor-threshold values. On the basis of these data, the aromas of both barley tea variants were successfully reconstituted with reference compounds. The calculation of odor-activity values (OAVs = concentration/odor-threshold value) and omission tests suggested 2-methoxyphenol (OAVs 69 and 160) and trans-isoeugenol (OAVs 1.4 and 31) as key compounds responsible for the stronger smoky note in the hulled barley tea. Further important odorants in the naked and hulled barley teas included 2-acetylpyrazine (OAVs 23 and 16), 2-acetyl-1-pyrroline (OAVs 19 and 16), and 3-methylbutanal (OAVs 12 and 15).