Aroma extract dilution analysis of distillates prepared by solvent extraction and solvent-assisted flavor evaporation distillation from white Alba truffle (WAT; Tuber magnatum pico) and Burgundy truffle (BT; Tuber uncinatum) revealed 20 odor-active regions in the flavor dilution (FD) factor range of 16–4096 in WAT and 25 in BT. The identification experiments in combination with the FD factors showed clear differences in the overall set of key odorants of both fungi. While 3-(methylthio)propanal (potato-like) followed by 2- and 3-methylbutanal (malty), 2,3-butanedione (buttery), and bis(methylthio)methane (garlic-like) showed the highest FD factors in WAT, 2,3-butanedione, phenylacetic acid (honey-like), and vanillin (vanilla-like) had the highest FD factors in BT. Odor activity values (OAVs, ratio of concentration to odor thresholds), which were calculated on the basis of quantitative data obtained by stable isotope dilution assays, of >1000 for bis(methylthio)methane, 3-methylbutanal, and 3,4-dihydro-2-(H)pyrrol (1-pyrroline) revealed they are key contributors to the aroma of WAT. In BT, 1-pyrroline and 2,3-butanedione showed the highest OAVs of 1530 and 1130, respectively. Aroma recombination experiments successfully mimicked the overall aroma profiles of both fungi when all odorants showing OAVs of >1 were combined. Omission experiments confirmed the amine-like and sperm-like smell of 1-pyrroline, identified for the first time as a key odorant in both truffle species.Keywords: 2,3-butanedione; 3,4-dihydro-2H-pyrrol; aroma extract dilution analysis; bis(methylthio)methane; stable isotope dilution assay; [13C4]-3,4-dihydro-2H-pyrrol; [2H8]bis(methylthio)methane;

An aroma distillate was prepared by solvent extraction and subsequent SAFE distillation from Italian vine-ripe tomatoes eliciting an intense overall aroma. Application of gc/olfactometry and the aroma extract dilution analysis revealed 44 odor-active compounds, 42 of which could be identified. The highest odor activity value of 2048 was established for the green, grassy (Z)-3-hexenal, the metallic smelling trans-4,5-epoxy-(E)-2-decenal, the potato-like 3-(methylthio)propanal, and the caramel-like 4-hydroxy-2,5-dimethyl-3(2H)-furanone. Of the further odorants, 13 compounds have previously not been reported as tomato odorants. Although most of these showed lower FD-factors, in particular, the coconut/dill-like smelling wine lactone ((3S,3aS,7aR)-3a,4,5,7a-tetrahydro-3,6-dimethylbenzofuran-2(3H)-one) appeared with a quite high FD factor. In addition, a fruity, almond-like odorant (6) with an FD factor of 1024 was detected. By application of high resolution mass spectrometry and polarity considerations, the structure of a methyl-2-ethoxytetrahydropyran isomer was suggested for 6. Four of the five possible isomers, the 3-methyl-, 4-methyl-, 5-methyl-, and 6-methyl-2-ethoxytetrahydropyran were synthesized and showed similar mass spectrometric patterns. However, these were excluded by their different retention indices. Although the synthesis of the remaining 2-methyl-2-ethoxytetrahydropyran resulted in only small yields, which were not sufficient for NMR measurements, this structure is very likely for 6. This compound was never reported as a food constituent before. Finally, quantitation of 23 odorants by stable isotope dilution assays allowed for the preparation of an aroma recombinate resembling the overall aroma of the tomatoes.Keywords: 2-methyl-2-ethoxytetrahydropyran; 3-methyl-2-ethoxytetrahydropyran; 5-methyl-2-ethoxytetrahydropyran; 6-methyl-2-ethoxytetrahydropyran; stable isotope dilution assay; wine lactone;

Application of aroma extract dilution analysis (AEDA) to an aroma distillate of blanched prawn meat (Litopenaeus vannamei) (BPM) revealed 40 odorants in the flavor dilution (FD) factor range from 4 to 1024. The highest FD factors were assigned to 2-acetyl-1-pyrroline, 3-(methylthio)propanal, (Z)-1,5-octadien-3-one, trans-4,5-epoxy-(E)-2-decenal, (E)-3-heptenoic acid, and 2-aminoacetophenone. To understand the influence of different processing conditions on odorant formation, fried prawn meat was investigated by means of AEDA in the same way, revealing 31 odorants with FD factors between 4 and 2048. Also, the highest FD factors were determined for 2-acetyl-1-pyrroline, 3-(methylthio)propanal, and (Z)-1,5-octadien-3-one, followed by 4-hydroxy-2,5-dimethyl-3(2H)-furanone, (E)-3-heptenoic acid, and 2-aminoacetophenone. As a source of the typical marine, sea breeze-like odor attribute of the seafood, 2,4,6-tribromoanisole was identified in raw prawn meat as one of the contributors. Additionally, the aroma of blanched prawn meat was compared to that of blanched Norway and American lobster meat, respectively (Nephrops norvegicus and Homarus americanus). Identification experiments revealed the same set of odorants, however, with differing FD factors. In particular, 3-hydroxy-4,5-dimethyl-2(5H)-furanone was found as the key aroma compound in blanched Norway lobster, whereas American lobster contained 3-methylindole with a high FD factor.Keywords: American lobster; aroma extract dilution analysis; blanched prawn meat; fried prawn meat; Norway lobster;

1-p-Menthene-8-thiol (1) has been discovered as the key odorant in grapefruit juice several decades ago and contributes to the overall odor of the fruit with an extremely low odor threshold of 0.000034 ng/L in air. This value is among the lowest odor thresholds ever reported for a food odorant. To check whether modifications in the structure of 1 would lead to changes in odor threshold and odor quality, 34 mercapto-containing p-menthane and 1-p-menthene derivatives as well as several aromatic and open-chain mercapto monoterpenoids were synthesized. Eighteen of them are reported for the first time in the literature, and their odor thresholds and odor qualities as well as analytical data are supplied. A comparison of the sensory data with those of 1 showed that hydrogenation of the double bond led to a clear increase in the odor threshold. Furthermore, moving the mercapto group into the ring always resulted in higher odor thresholds compared to thiols with a mercapto group in the side chains. Although all tertiary thiols always exhibited low odor thresholds, none of the 31 compounds reached the extremely low threshold of 1. Also, none of the synthesized mercapto monoterpenoids showed a similar odor quality resembling grapefruit. Although the saturated and aromatic analogues exhibited similar scents as 1, the aromas of the majority of the other compounds were described as sulfury, rubber-like, burned, soapy, or even mushroom-like. NMR and MS data as well as retention indices of the 23 newly reported sulfur-containing compounds might aid in future research to identify terpene-derived mercaptans possibly present in trace levels in foods.

Two rums differing in their overall aroma profile and price level (rum A, high price; rum B, low price) were analyzed by means of the Sensomics approach. Application of aroma extract dilution analysis (AEDA) on a distillate of volatiles prepared from rum A revealed 40 aroma-active compounds in the flavor dilution (FD) factor range from 8 to 2048. The identification experiments indicated cis-whiskey lactone, vanillin, decanoic acid, and 2- and 3-methylbutanol with the highest FD factors. The AEDA of a distillate prepared from rum B showed only 26 aroma-active compounds in the same FD factor range. Among them, in particular, ethyl butanoate, 1,1-diethoxyethane, ethyl (S)-2-methylbutanoate, and decanoic acid appeared with the highest FD factors. Thirty-seven compounds having at least an FD factor ≥32 in one of the two rums were quantitated using stable isotope dilution assays or enzyme kits (2 compounds). The calculation of odor activity values (OAVs; ratio of concentration to respective odor threshold) indicated ethanol, vanillin, ethyl (S)-2-methylbutanoate, and (E)-β-damascenone with the highest OAVs in rum A, whereas ethanol, 2,3-butanedione, 3-methylbutanal, and ethyl butanoate revealed the highest OAVs in rum B. Most compounds were present in similar concentrations in both rums, but significant differences were determined for vanillin, cis-whiskey lactone, and 4-allyl-2-methoxyphenol (all higher in rum A) and 3-methylbutanal, 2,3-butanedione, and ethyl butanoate (all higher in rum B). Finally, the aromas of both rums were successfully simulated by a recombinate using reference odorants in the same concentrations as they naturally occurred in the spirits.

The conversion of parent free amino acids into alcohols by an enzymatic deamination, decarboxylation, and reduction caused by microbial enzymes was first reported more than 100 years ago and is today known as the Ehrlich pathway. Because the chiral center at the carbon bearing the methyl group in l-isoleucine should not be prone to racemization during the reaction steps, the analysis of the enantiomeric distribution in 2-methylbutanal, 2-methylbutanol, and 2-methylbutanoic acid as well as in the compounds formed by secondary reactions, such as ethyl 2-methylbutanoate and 2-methylbutyl acetate, are an appropriate measure to follow the proposed degradation mechanism in the Ehrlich reaction. On the basis of a newly developed method for quantitation and chiral analysis, the enantiomers of the five metabolites were determined in a great number of fermented foods. Whereas 2-methylbutanol occurred as pure (S)-enantiomer in nearly all samples, a ratio of almost 1:1 of (S)- and (R)-2-methylbutanal was found. These data are not in agreement with the literature suggesting the formation of 2-methylbutanol by an enzymatic reduction of 2-methylbutanal. Also, the enantiomeric distribution in 2-methylbutanoic acid was closer to that in 2-methylbutanol than to that found in 2-methylbutanal, suggesting that also the acid is probably not formed by oxidation of the aldehyde as previously proposed. Additional model studies with (S)-2-methylbutanal did not show a racemization under the conditions of food production or during workup of the sample for volatile analysis. Therefore, the results establish that different mechanisms might be responsible for the formation of aldehydes and acids from the parent amino acids in the Ehrlich pathway.

An aroma extract dilution analysis (AEDA) carried out on the volatile fraction isolated by extraction/solvent-assisted flavor evaporation (SAFE) distillation from a commercial Amontillado sherry wine revealed 37 odor-active compounds with flavor dilution (FD) factors in the range of 16–4096. Among them, 2-phenylethanol (flowery, honey-like) and ethyl methylpropanoate (fruity) showed the highest FD factors, followed by ethyl (2S,3S)-2-hydroxy-3-methylpentanoate (fruity) reported for the first time in sherry wine. A total of 36 aroma-active compounds located by AEDA were then quantitated by a stable isotope dilution assay, and their odor activity values (OAVs; ratio of concentration to odor threshold) were calculated. The highest OAV was displayed by 1,1-diethoxyethane (2475; fruity), followed by 2- and 3-methylbutanals (574; malty) and methylpropanal (369; malty). Aroma reconstitution experiments and a comparative aroma profile analysis revealed that the entire orthonasal aroma profile of the Amontillado sherry wine could be closely mimicked.

In the volatile fraction isolated from a commercial Cognac by means of extraction/SAFE distillation, 39 odor-active areas were detected, among which (E)-β-damascenone showed the highest flavor dilution (FD) factor of 2048 followed by 2- and 3-methylbutanol, (S)-2-methylbutanol, 1,1-diethoxyethane, ethyl methylpropanoate, and ethyl (S)-2-methylbutanoate, as well as 4-hydroxy-3-methoxybenzaldehyde (vanilla-like) and 2-phenylethanol. The quantitation of 37 odorants by stable isotope dilution assays and a calculation of odor activity values (OAV; ratio of concentration to odor threshold) resulted in 34 odorants with OAVs > 1. Among them (E)-β-damascenone, methylpropanal, ethyl (S)-2-methylbutanoate, ethyl methylpropanoate, and ethyl 3-methylbutanoate together with ethanol were established as key contributors to the Cognac aroma. Finally, the overall aroma of the Cognac could be mimicked by an aroma recombinate consisting of these 34 key odorants on the basis of their natural concentrations in the Cognac using an odorless matrix to simulate the influence of the nonvolatile constituents. A comparison of the FD factors of the key odorants identified in a German brandy to those in the Cognac suggested the pair (E)-β-damascenone and ethyl pentanoate as indicators to differentiate various Cognacs from German, French, and Spanish brandies. This was confirmed by calculating a ratio of the concentrations of (E)-β-damascenone to ethyl pentanoate for 12 Cognacs and 7 brandies from Germany and 2 from France and Spain, respectively.

Acrolein (2-propenal) is classified as a foodborne toxicant and was shown to be present in significant amounts in heated edible oils. Up to now, its formation was mainly suggested to be from the glycerol part of triacylglycerides, although a clear influence of the unsaturation of the fatty acid moiety was also obvious in previous studies. To unequivocally clarify the role of the glycerol and the fatty acid parts in acrolein formation, two series of labeled triacylglycerides were synthesized: [13C3]-triacylglycerides of stearic, oleic, linoleic, and linolenic acid and [13C54]-triacylglycerides with labeled stearic, oleic, and linoleic acid, but with unlabeled glycerol. Heating of each of the seven intermediates singly in silicon oil and measurement of the formed amounts of labeled and unlabeled acrolein clearly proved the fatty acid backbone as the key precursor structure. Enzymatically synthesized pure linoleic acid and linolenic acid hydroperoxides were shown to be the key intermediates in acrolein formation, thus allowing the discussion of a radical-induced reaction pathway leading to the formation of the aldehyde. Surprisingly, although several oils contained high amounts of acrolein after heating, deep-fried foods themselves, such as donuts or French fries, were low in the aldehyde.

During thermal processing of foods, reducing carbohydrates and amino acids may form 1-amino-1-desoxyketoses named Amadori rearrangement products after the Italian chemist Mario Amadori. Although these compounds are transient intermediates of the Maillard reaction, they are often used as suitable markers to measure the extent of a thermal food processing, such as for spray-dried milk or dried fruits. Several methods are already available in the literature for their quantitation, but measurements are often done with external calibration without addressing losses during the workup procedure. To cope with this challenge, stable isotope dilution assays in combination with LC-MS/MS were developed for the glucose-derived Amadori products of the seven amino acids valine, leucine, isoleucine, phenylalanine, tyrosine, methionine, and histidine using the respective synthesized [13C6]-labeled isotopologues as internal standards. The quantitation of the analytes added to a model matrix showed a very good sensitivity with the lowest limits of detection for the Amadori compound of phenylalanine of 0.1 μg/kg starch and 0.2 μg/kg oil, respectively. Also, the standard deviation measured in, for example, wheat beer was only ±2% for this analyte. Application of the method to several foods showed the highest concentrations of the Amadori product of valine in unroasted cocoa (342 mg/kg) as well as in dried bell pepper (3460 mg/kg). In agreement with literature data, drying of foods led to the formation of Amadori products, whereas they were degraded during roasting of, for example, coffee or cocoa. The study presents for the first time results on concentrations of the Amadori compounds of tyrosine and histidine in foods.

Application of aroma extract dilution analysis (AEDA) on the volatiles isolated from a commercial Bavarian wheat beer (WB A) eliciting its typical aroma profile, best described by a clove-like, phenolic odor quality, revealed 36 odorants in the flavor dilution (FD) factor range from 16 to 4096. Among them, 2-methoxy-4-vinylphenol (clove-like) and 2-phenylethanol (flowery) showed the highest FD factors. AEDA of a second wheat beer (WB B), somewhat lacking the typical wheat beer odor note, revealed 32 odor-active components in the FD factor range from 32 to 8192. Among them, 2-phenylethanol, (E)-β-damascenone (cooked apple-like) and 3-methylbutanol (malty) were detected with the highest FD factors. Next, all odorants evaluated with an FD factor ≥32 were quantitated by stable isotope dilution assays in both beers, and the odor activity values (OAVs; ratio of concentration to odor threshold) were calculated. Thereby, ethanol, (E)-β-damascenone, 3-methylbutyl acetate, ethyl methylpropanoate, and ethyl butanoate showed the highest OAVs in WB A, followed by acetaldehyde, 3-methylbutanol, and dimethyl sulfide. In WB B, ethanol, (E)-β-damascenone, ethyl methylpropanoate, ethyl butanoate, and 3-methylbutyl acetate showed the highest OAVs. Whereas most aroma compounds were present in the same order of magnitude in both beer samples, in particular, 2-methoxy-4-vinylphenol and 4-vinylphenol (smoky, leather-like) were by factors of 13 and 15, respectively, higher in WB A. For the first time, the overall aroma of wheat beer (WB A) was successfully simulated on the basis of 27 reference compounds in their natural concentrations using water/ethanol (95:5; v/v) as the matrix.

Due to the high number of double bonds, ω-3-polyunsaturated fatty acids such as eicosapentaenoic aid (EPA) or docosahexaenoic acid (DHA) are prone to rapid oxidation, leading to the formation of intense taints often described as “fishy”. To clarify the compounds responsible for such off-flavors, EPA, DHA, and α-linolenic acid (ALA) were oxidized singly either in the presence of copper ions or in the presence of lipoxygenase. The autoxidation of EPA and DHA led to a mixture of odorants eliciting an overall fishy odor quality, whereas neither the oxidation of ALA by copper ions nor that by lipoxygenase led to an unpleasant odor. Application of aroma extract dilution analysis (AEDA) on the volatiles generated by autoxidation of EPA revealed trans-4,5-epoxy-(E,Z)-2,7-decadienal, identified for the first time as fatty acid degradation product, (Z)-1,5-octadien-3-one, (Z)-3-hexenal, (Z,Z)-2,5-octadienal, (Z,Z)-3,6-nonadienal, and (E,E,Z)-2,4,6-nonatrienal with the highest flavor dilution (FD) factors. The autoxidation as well as the enzymatic oxidation of all three acids led to the same odorants, but with different FD factors depending on the acid and/or the type of oxidation applied. Thus, the results suggested that a defined ratio of a few odorants is needed to generate a fishy off-flavor.

The aroma compounds in two commercial Bartlett pear brandies clearly differing in their overall aroma profiles were detected in the volatile fractions by the aroma extract dilution analysis. In brandy A eliciting the more intense pear-like, fruity aroma, ethyl (S)-2-methylbutanoate, (E)-β-damascenone, 1,1-diethoxyethane, 2- and 3-methylbutanol, (S)-2- and 3-methylbutanoic acid, and 2-phenylethanol were found with the highest Flavor Dilution (FD) factors. In brandy B judged to have a weaker overall aroma, also (E)-β-damascenone, ethyl (S)-2-methylbutanoate, and 2-phenylethanol revealed high FD factors, while many odorants showed lower FD factors. Fourty-four odor-active compounds were quantitated by stable isotope dilution assays, and the odor activity values (OAVs; ratio of concentrations to odor thresholds) confirmed (E)-β-damascenone and ethyl (S)-2-methylbutanoate as important aroma compounds in brandy A, while the OAVs of most odorants were much lower in brandy B. By aroma recombination studies, the aromas of both brandies could be matched using reference odorants in the same concentrations as they occurred in the spirits. In 15 commercial Bartlett pear brandies ethyl (E,Z)-2,4-decadienoate and (E,E)-2,4-decadienoate eliciting a pear-like aroma showed a reasonable correlation of their concentrations with the overall aroma quality.

Application of the aroma extract dilution analysis on a distillate prepared from an authentic Styrian pumpkin seed oil followed by identification experiments led to the characterization of 47 odor-active compounds in the flavor dilution (FD) factor range of 8–8192 among which 2-acetyl-1-pyrroline (roasty, popcorn-like), 2-propionyl-1-pyrroline (roasty, popcorn-like), 2-methoxy-4-vinylphenol (clove-like), and phenylacetaldehyde (honey-like) showed the highest FD factors. Among the set of key odorants, 2-propionyl-1-pyrroline and another 20 odorants were identified for the first time as constituents of pumpkin seed oil. To evaluate the aroma contribution in more detail, 31 aroma compounds showing the highest FD factors were quantitated by means of stable isotope dilution assays. On the basis of the quantitative data and odor thresholds determined in sunflower oil, odor activity values (OAV; ratio of concentration to odor threshold) were calculated, and 26 aroma compounds were found to have an OAV above 1. Among them, methanethiol (sulfury), 2-methylbutanal (malty), 3-methylbutanal (malty), and 2,3-diethyl-5-methylpyrazine (roasted potato) reached the highest OAVs. Sensory evaluation of an aroma recombinate prepared by mixing the 31 key odorants in the concentrations as determined in the oil revealed that the aroma of Styrian pumpkin seed oil could be closely mimicked. Quantitation of 11 key odorants in three commercial pumpkin seed oil revealed clear differences in the concentrations of distinct odorants, which were correlated with the overall aroma profile of the oils.

By application of aroma extract dilution analysis (AEDA) on the volatile fraction isolated by solvent extraction and solvent-assisted flavor evaporation (SAFE) from unifloral rape honey harvested in July 2009, 28 odor-active areas could be detected within a flavor dilution factor (FD) range of 4–2048. The highest FD factors were found for (E)-β-damascenone (cooked apple-like), phenylacetic acid (honey-like), 4-methoxybenzaldehyde (aniseed-like), 3-phenylpropanoic acid (flowery, waxy), and 2-methoxy-4-vinylphenol (clove-like). Twenty-three odorants were then quantitated by application of stable isotope dilution assays, and their odor activity values (OAV, ratio of concentration to odor threshold) were calculated on the basis of newly determined odor thresholds in an aqueous fructose–glucose solution. The highest OAVs were calculated for (E)-β-damascenone, 3-phenylpropanoic acid, phenylacetic acid, dimethyl trisulfide, and phenylacetaldehyde. Quantitative measurements on a rape honey produced in 2011 confirmed the results. A model mixture containing the 12 odorants showing an OAV ≥ 1 at the same concentrations as they occurred in the rape honey was able to mimick the aroma impression of the original honey. The characterization of the key odorants in rape flowers from the same field suggested 3-phenylpropanoic acid, phenylacetic acid, and three further odorants to be transferred via the bees into the honey.

Microbial amino acid metabolism may lead to substantial amounts of biogenic amines in either spontaneously fermented or spoiled foods. For products manufactured with starter cultures, it has been suggested that certain strains may produce higher amounts of such amines than others; however, to support efforts of food manufacturers in mitigating amine formation, reliable methods for amine quantitation are needed. Using 10 isotopically labeled biogenic amines as the internal standards, stable isotope dilution assays were developed for the quantitation of 12 biogenic amines and of the 2 polyamines, spermine and spermidine, in one LC-MS/MS run. Application of the method to several foods revealed high concentrations of, for example, tyramine and putrescine in salami and fermented cabbage, whereas histamine was highest in Parmesan cheese and fermented cabbage. On the other hand, ethanolamine was highest in red wine and Parmesan cheese. The results suggest that different amino acid decarboxylases are active in the respective foods depending on the microorganisms present. The polyamine spermine was highest in salami and tuna.

Quantification of aroma compounds in an orange juice reconstituted from concentrate which had been stored at 37 °C for 4 weeks (forced storage) revealed an increase in the concentrations of, in particular, dimethyl sulphide, 2-methoxy-4-vinylphenol, α-terpineol, and 4-hydroxy-2,5-dimethyl-3(2H)-furanone (4-HDF) in comparison with the same orange juice before storage. On the other hand, clearly lower concentrations were found for octanal, decanal, (R)-α-pinene, linalool, and (E)-β-damascenone after storage, while the concentrations of vanillin and carvone remained nearly constant. Similar results were found for the same aroma compounds after storage of the orange juice at 20 °C for 1 year. Sensory experiments corroborated the importance of 2-methoxy-4-vinylphenol and dimethyl sulphide for the typical stale off-flavour of the stored orange juice, while the omission of e.g., α-terpineol in model mixtures could not be detected. Under both storage conditions (37 °C for 4 weeks or 20 °C for 1 year), the breakthrough odour thresholds of α-terpineol and 4-HDF were not reached, while the concentrations of dimethyl sulphide and 2-methoxy-4-vinylphenol clearly exceeded their breakthrough odour thresholds, thus confirming the crucial role of these odorants for the off-flavour of stored orange juice from concentrate. In addition, changes in the concentrations of selected orange juice odorants at various temperatures and times were investigated.

By means of stable isotope dilution assays (SIDA), 26 odor-active compounds, previously characterized by GC−olfactometry (GC-O), were quantitated in raw peanuts, and the concentrations of 38 odorants were determined in pan-roasted peanut meal. On the basis of the quantitative data and odor thresholds determined in vegetable oil, the odor activity values (OAVs) of the most important aroma compounds in raw as well as in pan-roasted peanut meal were calculated. 3-Isopropyl-2-methoxypyrazine, acetic acid, and 3-(methylthio)propanal showed the highest OAVs in raw peanuts, whereas methanethiol, 2,3-pentanedione, 3-(methylthio)propanal, and 2- and 3-methylbutanal as well as the intensely popcorn-like smelling 2-acetyl-1-pyrroline revealed the highest OAV in the pan-roasted peanut meal. Aroma recombination studies confirmed the importance, in particular, of methanethiol and of lipid degradation products in the characteristic aroma of the freshly roasted peanut material. To evaluate additive effects on the overall aroma, the concentrations of eight pyrazines, previously not detected by GC-O among the odor-active volatiles, were additionally quantitated in the pan-roasted peanut meal. A sensory experiment in which the eight pyrazines were added to the recombinate clearly revealed that these volatiles did not show an impact on the overall aroma. Finally, selected odorants were quantitated in commercial peanut products to confirm their important role in peanut aroma.

The potent odorant dimethyl sulfide (1), showing a low odor threshold of 0.12 μg/L in water, is known to contribute to the aromas of various foods. Its cabbage-like odor plays an important role, particularly, in cooked vegetables, such as cabbage, celery, or asparagus. On the other hand, in fruit juices or beer, 1 may generate off-flavors. S-Methylmethionine (2) has previously been characterized as precursor of 1 during thermal processing, and several methods for its quantitation have been proposed. Using deuterium-labeled 2 as the internal standard, a stable isotope dilution assay (SIDA) using LC-MS/MS was developed for the fast quantitation of 2 in vegetables and malt. Application of the method to different foods revealed amounts between 2.8 mg (fresh tomatoes) and 176 mg (celery) of 2 per kilogram. To correlate the amount of 1 formed upon processing with the amounts of 2 present in the raw material, 1 was quantified before and after a thermal treatment of the same raw materials by a SIDA. Concentrations between 1.1 mg/kg (fresh tomatoes) and 26 mg/kg (celery) were determined in the processed samples. The quantitation of 2 during steeping, germination, and malting of barley, and a correlation of the data with the amounts of 1 formed after thermal treatment of the malt, resulted in yields between 24 and 27 mol % calculated on the basis of the amounts of 2. The results suggested that the extent of the formation of 1 can be predicted, for example, in plant materials, from the amount of 2 present in the raw foods.

Seventeen aroma-active volatiles, previously identified with high flavor dilution factors in fresh, pink Colombian guavas (Psidium guajava L.), were quantified by stable isotope dilution assays. On the basis of the quantitative data and odor thresholds in water, odor activity values (OAV; ratio of concentration to odor threshold) were calculated. High OAVs were determined for the green, grassy smelling (Z)-3-hexenal and the grapefruit-like smelling 3-sulfanyl-1-hexanol followed by 3-sulfanylhexyl acetate (black currant-like), hexanal (green, grassy), ethyl butanoate (fruity), acetaldehyde (fresh, pungent), trans-4,5-epoxy-(E)-2-decenal (metallic), 4-hydroxy-2,5-dimethyl-3(2H)-furanone (caramel, sweet), cinnamyl alcohol (floral), methyl (2S,3S)-2-hydroxy-3-methylpentanoate (fruity), cinnamyl acetate (floral), methional (cooked potato-like), and 3-hydroxy-4,5-dimethyl-2(5H)-furanone (seasoning-like). Studies on the time course of odorant formation in guava puree or cubes, respectively, showed that (Z)-3-hexenal was hardly present in the intact fruits, but was formed very quickly during crushing. The aroma of fresh guava fruit cubes, which showed a very balanced aroma profile, was successfully mimicked in a reconstitute consisting of 13 odorants in their naturally occurring concentrations. Omission tests, in which single odorants were omitted from the entire aroma reconstitute, revealed (Z)-3-hexenal, 3-sulfanyl-1-hexanol, 4-hydroxy-2,5-dimethyl-3(2H)-furanone, 3-sulfanylhexyl acetate, hexanal, ethyl butanoate, cinnamyl acetate, and methional as the key aroma compounds of pink guavas.

Application of an aroma extract dilution analysis on the entire volatile fraction isolated from an orange juice freshly reconstituted from concentrate revealed 40 odour-active constituents in the flavour dilution (FD) factor range of 4–2,048. Among them, ethyl butanoate and linalool showed the highest FD factor of 2,048, followed by octanal with an FD factor of 512. Thirty-six of the 40 odour-active compounds detected could be identified, all of which have previously been reported as volatile constituents of various orange juices. Quantification of 17 key odorants by stable isotope dilution assays followed by a calculation of odour activity values (OAVs) on the basis of odour thresholds in water or citrate buffer (pH 3.8), respectively, revealed the following most important odorants in the overall aroma of the freshly reconstituted juice: (R/S)-linalool, (R)-limonene and (S)-ethyl 2-methylbutanoate with the highest OAVs (>1,000) followed by octanal, (R)-α-pinene, ethyl butanoate, myrcene, acetaldehyde, decanal and (E)-β-damascenone with OAVs > 100. A model mixture containing all 14 aroma compounds with OAVs > 1 in their actual concentrations in the juice showed a good similarity with the aroma of the original orange juice under investigation, thus corroborating that the key odorants of a freshly reconstituted orange juice were characterised for the first time.

Application of the aroma extract dilution analysis (AEDA) on the volatile fraction carefully isolated from an American Bourbon whisky revealed 45 odor-active areas in the flavor dilution (FD) factor range of 32−4096 among which (E)-β-damascenone and δ-nonalactone showed the highest FD factors of 4096 and 2048, respectively. With FD factors of 1024, (3S,4S)-cis-whiskylactone, γ-decalactone, 4-allyl-2-methoxyphenol (eugenol), and 4-hydroxy-3-methoxy-benzaldehyde (vanillin) additionally contributed to the overall vanilla-like, fruity, and smoky aroma note of the spirit. Application of GC-Olfactometry on the headspace above the whisky revealed 23 aroma-active odorants among which 3-methylbutanal, ethanol, and 2-methylbutanal were identified as additional important aroma compounds. Compared to published data on volatile constituents in whisky, besides ranking the whisky odorants on the basis of their odor potency, 13 aroma compounds were newly identified in this study: ethyl (S)-2-methylbutanoate, (E)-2-heptenal, (E,E)-2,4-nonadienal, (E)-2-decenal, (E,E)-2,4-decadienal, 2-isopropyl-3-methoxypyrazine, ethyl phenylacetate, 4-methyl acetophenone, α-damascone, 2-phenylethyl propanoate, 3-hydroxy-4,5-dimethyl-2(5H)-furanone, trans-ethyl cinnamate, and (Z)-6-dodeceno-γ-lactone.

Application of a comparative aroma extraction dilution analysis on unroasted and roasted Criollo cocoa beans revealed 42 aroma compounds in the flavor dilution (FD) factor range of 1−4096 for the unroasted and 4−8192 for the roasted cocoa beans. While the same compounds were present in the unroasted and roasted cocoa beans, respectively, these clearly differed in their intensity. For example, 2- and 3-methylbutanoic acid (rancid) and acetic acid (sour) showed the highest FD factors in the unroasted beans, while 3-methylbutanal (malty), 4-hydroxy-2,5-dimethyl-3(2H)-furanone (caramel-like), and 2- and 3-methylbutanoic acid (sweaty) were detected with the highest FD factors in the roasted seeds. Quantitation of 30 odorants by means of stable isotope dilution assays followed by a calculation of odor activity values (ratio of the concentration/odor threshold) revealed concentrations above the odor threshold for 22 compounds in the unroasted and 27 compounds in the roasted cocoa beans, respectively. In particular, a strong increase in the concentrations of the Strecker aldehydes 3-methylbutanal and phenylacetaldehyde as well as 4-hydroxy-2,5-dimethyl-3(2H)-furanone was measured, suggesting that these odorants should contribute most to the changes in the overall aroma after roasting. Various compounds contributing to the aroma of roasted cocoa beans, such as 3-methylbutanoic acid, ethyl 2-methylbutanoate, and 2-phenylethanol, were already present in unroasted, fermented cocoa beans and were not increased during roasting.

Application of an aroma extract dilution analysis on an aroma distillate prepared from organically grown, raw West-African peanuts (Cameroon) revealed 36 odor-active areas in the flavor dilution (FD) factor range of 1 to 2048. The identification experiments, which were all performed by using the respective reference chemicals, revealed 2-isopropyl-3-methoxypyrazine (earthy, pea-like), 2-isobutyl-3-methoxypyrazine (bell pepper-like, earthy), and trans-4,5-epoxy-(E)-2-decenal (metallic) with the highest FD factors among the 36 aroma compounds identified. The two last mentioned odorants and another set of 22 further odorants were identified for the first time in raw peanuts. A comparative aroma extract dilution analysis applied on distillates prepared from either the raw peanuts or ground peanut meal roasted in a pan showed 52 odor-active areas in the FD factor range of 8 to 2048 in the roasted nut material. The identification experiments in combination with the FD factors revealed that among them, 2-acetyl-1-pyrroline and 4-hydroxy-2,5-dimethyl-3-(2H)-furanone showed the most significant contribution to the overall aroma, followed by 1-octen-3-one, 2-isopropyl-3-methoxypyrazine, (E,E)-2,4-decadienal, and trans-4,5-epoxy-(E)-2-decenal. As a further result, 20 aroma compounds were newly identified in roasted peanuts, such as 2-propionyl-1-pyrroline and 2-acetyltetrahydropyridine (both popcorn-like). In particular, 2-acetyl-1-pyrroline and 4-hydroxy-2,5-dimethyl-3(2H)-furanone showed the most pronounced increase after roasting.

Thirty-one of the 45 odor-active compounds previously identified by us in an American Bourbon whisky were quantified by stable isotope dilution assays. Also for this purpose, new synthetic pathways were developed for the synthesis of the deuterium-labeled whisky lactone as well as for γ-nona- and γ-decalactone. To obtain the odor activity values (OAVs), the concentrations measured were divided by the odor thresholds of the odorants determined in water/ethanol (6:4 by vol.). Twenty-six aroma compounds showed OAVs >1, among which ethanol, ethyl (S)-2-methylbutanoate, 3-methylbutanal, 4-hydroxy-3-methoxybenzaldehyde, (E)-β-damascenone, ethyl hexanoate, ethyl butanoate, ethyl octanoate, 2-methylpropanal, (3S,4S)-cis-whiskylactone, (E,E)-2,4-decadienal, 4-allyl-2-methoxyphenol, ethyl-3-methylbutanoate, and ethyl 2-methylpropanoate showed the highest values. The overall aroma of the Bourbon whisky could be mimicked by an aroma recombinate consisting of the 26 key odorants in their actual concentrations in whisky using water/ethanol (6:4 by vol.) as the matrix. Omission experiments corroborated the importance of, in particular, 4-hydroxy-3-methoxybenzaldehyde, (3S,4S)-cis-whiskylactone, ethanol, and the entire group of esters for the overall aroma of the Bourbon whisky.

The volatiles present in fresh, pink-fleshed Colombian guavas (Psidium guajava, L.), variety regional rojo, were carefully isolated by solvent extraction followed by solvent-assisted flavor evaporation, and the aroma-active areas in the gas chromatogram were screened by application of the aroma extract dilution analysis. The results of the identification experiments in combination with the FD factors revealed 4-methoxy-2,5-dimethyl-3(2H)-furanone, 4-hydroxy-2,5-dimethyl-3(2H)-furanone, 3-sulfanylhexyl acetate, and 3-sulfanyl-1-hexanol followed by 3-hydroxy-4,5-dimethyl-2(5H)-furanone, (Z)-3-hexenal, trans-4,5-epoxy-(E)-2-decenal, cinnamyl alcohol, ethyl butanoate, hexanal, methional, and cinnamyl acetate as important aroma contributors. Enantioselective gas chromatography revealed an enantiomeric distribution close to the racemate in 3-sulfanylhexyl acetate as well as in 3-sulfanyl-1-hexanol. In addition, two fruity smelling diastereomeric methyl 2-hydroxy-3-methylpentanoates were identified as the (R,S)- and the (S,S)-isomers, whereas the (S,R)- and (R,R)-isomers were absent. Seven odorants were identified for the first time in guavas, among them 3-sulfanylhexyl acetate, 3-sulfanyl-1-hexanol, 3-hydroxy-4,5-dimethyl-2(5H)-furanone, trans-4,5-epoxy-(E)-2-decenal, and methional were the most odor-active.

Isolation of the volatile fraction from the peel of Pontianak orange (Citrus nobilis var. Lour. microcarpa Hassk.) by a careful solvent extraction/vacuum distillation process followed by application of an aroma extract dilution analysis (AEDA) revealed 32 odour-active compounds in the flavour dilution (FD)-factor range of 4–2048, 26 of which could be identified. On the basis of high FD-factors, (R)/(S)-linalool, myrcene, (R)-limonene, and 1,8-cineole were characterised as the most potent odorants, followed by octanal, (E,E)-2,4-decadienal, nerol, (E)-2-dodecenal, geraniol, and (E,E)-2,4-nonadienal. In addition, one odorant resembling the characteristic sulphurous, resinous aroma of the Pontianak peel oil was detected with a quite high FD-factor of 128. By mass spectrometry followed by synthesis, 1-phenylethanethiol exhibiting an intense sulphurous, resinous smell at the very low odour threshold of 0.005 ng/L in air, was found to be responsible for the odour impression detected in the extract. 1-Phenylethanethiol occurring as a mixture of the (R)-(76%) and the (S)-enantiomer (24%) has previously not been reported as odorant in foods. Application of static headspace aroma dilution analysis (SHA) on Pontianak peel revealed the green, grassy smelling odour-active compounds hexanal and (Z)-3-hexenal as further important odorants in the headspace above the peels.

Application of a comparative aroma extract dilution analysis on the volatiles isolated from five different hops (Hallertau Perle, Hallertau Hersbrucker Spät, Slowenian Golding, Hallertau Smaragd, US Cascade) revealed linalool and myrcene with the highest Flavour Dilution (FD)-factors in all varieties, followed by 2-isopropyl-3-methoxypyrazine, 3-methylbutanoic acid and geraniol. Some odourants, however, showed high FD-factors only in certain varieties, for example, (5Z)-octa-1,5-dien-3-one and germacrene B in Hersbrucker Spät, (3E,5Z)-undeca-1,3,5-triene in Hersbrucker Spät and Cascade and nonanal in Cascade. The overall odour profile of the Cascade sample clearly differed from the other varietes, and was dominated by a black currant like odour note. The identification experiments revealed 4-methyl-4-sulfanylpentan-2-one, so far unknown as hop constituent, as key contributor to this odour. In addition, an odour-active undecatetraene was present, in particular, in Perle and Cascade. Synthesis and structural assignment of the four stereoisomers of (3E)-undeca-1,3,5,9-tetraene allowed the identification of the fresh, pineapple-like smelling compound as (3E,5Z,9E)-undeca-1,3,5,9-tetraene. Among the four isomers synthesised, this compound showed by far the lowest odour threshold of 0.01 ng/L in air.

Application of an Aroma Extract Dilution Analysis (AEDA) on distillates prepared from two UHT-milks either packed in polyethylene (PE-M) or glass (G-M), respectively, revealed δ-decalactone and γ-dodecalactone as the most odour-active constituents among the 37 odour-active constituents in the PE-M. δ-Decalactone was also the most odour-active among the 32 odorants detected in G-M, followed by δ-octalactone, γ-dodecalactone, (Z)-6-γ-dodecenolactone and vanillin. To detect a possible adsorption of odorants at the packaging, the AEDA was also applied on solvent extracts obtained from both, the glass and the polyethylene bottles, in which the two milks were supplied. The results confirmed the high tendency of polyethylene to adsorb odorants, while the affinity of the aroma compounds to glass were weak. A comparative quantification of selected key odorants in the polyethylene packaging as well as in the milk, allowed the calculation of adsorption ratios. The results demonstrated a strong influence of the chemical structure on the extent of adsorption. For instance, aldehydes, such as (E)-2-nonenal and (E,Z)-2,6-nonadienal, had a much higher affinity to polyethylene than lactones and acids. The results indicated that the overall aroma of UHT-milk packed in polyethylene can be changed during storage by adsorption as well as by permeation processes of milk aroma compounds to and through polyethylene packaging.