FLAVOUR AND FRAGRANCE JOURNAL, VOL. 9.

147-158 (1994)

Review

Determination of Potent Odourants in Foods by Aroma

Extract Dilution Analysis (AEDA) and Calculation of Odour
Activity Values (OAVs)
Werner Grosch
Deutsche Forschungsanstalt fur Lehensmittelchemie, Lichtenbergsirasse 4, 0-85748 Garching, Germany

The potency of odourants in foods is determined by aroma extract dilution analysis (AEDA) and by the
calculation of odour activity values (OAVs). The application of these methods to olive oil, butter, Swiss cheese,
heated beef, bread, beer, green tea, dill herb and to the off-flavoursformed in lemon oil, soybean oil and boiled
trout is reviewed. The results allow the conclusion that only a small fraction of the complex mixture of volatiles
causes the characteristic odour of a distinct food.
KEY WORDS Odourant Aroma extract dilution analysis Odour activity value
Gas chromatography-olfactometry Review

INTRODUCTION tectors in which the peak areas are proportional

There are indications that only a small fraction of
the complex mixture of volatiles occurring in
'
foods causes the overall odour. In consequence,
the distinction of the more potent odourants from
the volatiles having low or no odour activity is the
first task to be solved in flavour analysis.
As recently reviewed,' v2 two techniques,
Charm analysis and aroma extract dilution analy-
sis (AEDA), are suitable to screen the potent
odourants on which the identification experi-
ments are focused. In both procedures an extract
obtained from the food is diluted, usually as a
series of 1:l or 1:2 dilutions, and each dilution is
analysed by gas chromatography-olfactometry
(GC-0). In the case t f AEDA, the result is
expressed as flavour dilution (FD)-factor,' which
is the ratio of the concentration of the odourant in
the initial extract to its concentration in the most
dilute extract in which odour was detected by
GC-0. Consequently, the FD-factor is a relative
measure and is proportional to the odour activity
value (OAV, ratio of concentration to odour
threshold) of the compound in air.
Charm analysis constructs chromatographic
peaks similar to those produced by chemical de-
to the amount of chemical in the extract.' The
primary difference between the two methods is
that Charm analysis measures the dilution value
over the entire time the compounds elute, whereas
AEDA simply determined the maximum dilution
value detected.'
Charm analysis and AEDA are both screening
procedures,' as the results are not corrected for
the losses of odourants during the isolation pro-
cedure. Even the errors due to a lack in experience
of the persons performing the dilution experi-
ment, e.g. the perception of gaps,3 have to be
eliminated. Hence, to indicate which of the com-
pounds revealed by the dilution procedures
actually contributes to the aroma of a particular
food, quantification of the odourants with high
FD-factors and calculation of their OAVs are the
next steps in the analytical procedure. Odourants
with high OAVs are important contributors to the
characteristic flavours or off-flavours and are
suitable as indicator substances for the objective
determination of flavour differences in foods.
Since its publication4 in 1987 AEDA has been
applied to a number of foods. In the following,
only those results are reviewed in which the sim-
plifications implicit in AEDA are corrected for by

CCC 0882-5734/94/O40147- 12 Received29 January 1992

0 1994 by John Wiley & Sons, Ltd. Accepted 5 April 1994
148 W. GROSCH

the quantification of the level of the odourants the highest FD-factors in the samples A and B,
and by the calculation of the OAVs. respectively.
Those compounds with an FD-factor of at least
2' in the two oil samples were quantified by stable
OLIVE OIL isotope dilution assays and their OAVs were
calculated6 to evaluate the odourants responsible
The odour of virgin olive oil can vary greatly. for the difference in the odour profiles shown in
Table 1 shows two oil samples of which A smelled Table 1. The results (Table 2) confirmed the
fruity, green, fatty and B strongly blackcurrant- character-impact odourants evaluated by the
like. comparative AEDA of the two oil samples. In
A comparative AEDA was performed' to sample A the OAV of (Z)-hex-3-enal surpassed
identify the compounds responsible for the differ- by far the OAVs of the other odourants thus
ence in the odour profiles. In the FD-factor range indicating that it caused the green odour note
24 to 2", 33 of the 36 odourants screened by predominating in the odour profile. Although the
AEDA were identified. (Z)-Hex-3-enal, smelling concentration of 4-methoxy-2-methyl-2-butane-
green, leaf-like, and 4-methoxy-Z-methyl-2- thiol amounted only to 1.8pg/kg (Table 2), it had
butanethiol, smelling blackcurrant-like, showed a great impact on the odour of sample B owing to
its low odour threshold of 45ng/kg6 and caused
the blackcurrant odour note in B.
Table 1. Odour profiles of two virgin olive oil samples' The fruity odour notes in samples A and B
Olive oil were caused by the esters nos 5 to 7 (Table 2 ) of
Odour quality Sample A Sample B which the OAVs were much higher in B than in
A. (Z)-Hex-3-enyl acetate (no. 8) was by far the
Mean panel score of odour intensity" major compound in the fraction of esters listed in
Fruity 1.1 1.6
Green 1.5 0 Table 2. However, the odour threshold of no. 8 in
Blackcurrant-like 0 2.5 the oils was higher6 by a factor of lo4 than that of
Fatty 1.7 0.6 the esters nos 5 to 7. Therefore, the OAV of no. 8
Floral 0.3 0.3 was lower in sample B, and this suggests that the
Spicy 0.5 0.5 contribution of no. 8 to the fruity odour note of R
______~

"Rating scale of the odour intensity: 0, odour quality is lack- was relatively small.
ing; 1, bland: 2, moderate; 3, strong. The OAV of (Z)-hex-3-enal in sample B

Table 2. Potent odourants of two virgin olive oil samples'."

Sample A Sample B
No. Compound n" (pg/kg) OAV" na (pg/kg) OAV"
I Acetic acid 5 2449 2.3 7 6683 6.4
3- 3-Methylbutanol <4 116 <l 7 904 4
3 2-Phenylethanol 5 363 3.2 7 345 3.1
4 (Z)-Hex-3-enol 7 662 <I 7 796 <I
5 Ethyl 2-methylbutyrate 6 2.0 2.7 8 14 19
6 Ethyl isobutyrate 4 1.8 2.4 7 7.9 I1
7 Ethyl cyclohexanoate 6 1.2 3.2 10 4.3 11
8 (Z)-Hex-3-enyl acetate 8 3212 4.3 8 3383 4.5
9 4-Methoxy-2-methyl-2-butanethiol <4 <O, 1 <I 10 1.8 40
10 Hexanal 9 1274 17 7 388 5.3
11 (Q-Hex-2-enal 7 4296 3.8 4 365 <1
12 (Z)-Hex-3-enal 11 325 116 7 53 19
13 (Z)-Non-2-enal 9 14 23 8 10 17
14 (E,E)-Deca-2,4-dienaI 8 224 5.5 7 111 2.7
"Exponent of the FD-factor 2".
bodour activity values (OAVs) were calculated by dividing the concentrations of the odourants by
their retronasal odour thresholds in sunflower oil.
 AROMA EXTRACT DILUTION ANALYSIS 149

amounted to 19 (Table 2), but there was no per- Table 4. Potent odourants revealed in butter sample B by
ceptible reen note (Table 1). It was, therefore, AEDA~
assumed f e that the odour of (Z)-hex-3-enal in B
No. Compound
was masked by the intense odours of the thiol
no. 9 and the esters nos 5 to 7. 1 D iacet y I
A study on an olive oil sample smelling intensely 2 Pent-I-en-3-one
3 Hexanal
fatty' showed that in addition to (Z)-non-2-ena17 4 Oct-1-en-3-one
octanal, and oct-1-en-3-one might be responsible 5 Unknown
for the fatty odour notes on the basis of their high 6 (Z,Z)-Nona-3,6-dienal
OAVs. 7 (Z)-Non-2-enal
8 (E)-Non-3-enal
9 (E,E)-Nona-2,4-dienal
10 (E,E)-Dcca-2,4-dienaI
BUITER 11 y-Octalactone
12 trans-4,5-Epoxy-( E)-dec-2-enal
In a study on butter flavours,8 an Irish sour cream 13 Skatole
14 &-Decalactone
butter (sample A), showing the most intense 15 (Z)-Dodec-6-eno-y-lactone
buttery, creamy and sweet odour of the five sam- 16 Acetic acid
ples listed in Table 3, was chosen for AEDA. The 17 Butanoic acid
identification experiments revealed 18 potent 18 Hexanoic acid
odourants (Table 4) which, with the exception of
nos 6, 12 and 18, agreed with those of butter oil.'
To gain an insight into the flavour differences
of the butter samples (Table 3), five compounds OAVs of the five odourants in these samples. On
revealed by the AEDA as important butter the other hand, this note was weak in sample C in
odourants were quantified, and their OAVs were which the OAV of diacetyl was lower and in the
calculated. A comparison of Tables 3 and 5 allows same range as the OAVs of &decalactone and
the conclusion that the buttery note in the odour butanoic acid. Furthermore, the buttery note was
profiles of the samples A and B was caused by the lacking in sample D which was free of diacetyl.
buttery-smelling diacetyl, showing the highest Butanoic acid was the character impact odourant
of sample E. Its OAV was much higher than the
Table 3. Odour profiles of five butter samples" OAVs of the other odourants. Consequently, its
___ ~ ~ _ _ _ sweaty, rancid odour predominated the odour
Sample Odour quality Intensity" profile of E. Compared to diacetyl, 6-decalactone
A buttery, creamy, sweet 3
and butanoic acid, the OAVs of (Z)-dodec-6-eno-
B typical butter-like, sweet 2-3 y-lactone and hexanoic acid were low in the butter
C mild, weakly buttery, sour 1-2 samples indicating that these odourants did not
D slightly sour, mild 1 significantly contribute to the flavour of butter.
E rancid, like butanoic acid 3 To establish the character impact odourants of
"Rating scale of the overall odour intensity: 1, weak; 2, sample B, sunflower oil was spiked with diacetyl,
medium; 3, strong. &decalactone and butanoic acid in various com-

Table 5. Concentration and OAVs of five odourants in the butter samples A to E8

~~~ ~

Sample A Sample B Sample C Sample D Sample E

Compound (mg/kg) OAV" (mg/kg) OAV" (mglkg) OAV" (mg/kg) OAV" (mg/kg) OAV"

Diacetyl 0.62 138 0.34 76 0.11 24 <0.01 <2 0.32 71

&Decalactone 5.0 42 4.9 41 3.06 26 3.80 32 2.15 18
Butanoic acid 4.48 33 3.63 27 2.67 20 2.48 19 94.5 704
(Z)-Dodec-6-eno-y-lactone 0.26 1 0.16 <1 0.078 <I 0.11 <1 0.057 <1
Hexanoic acid 1.84 <I 1.87 <I 1.05 c1 0.99 <1 16.8 3

"The odour activity value (OAV) was calculated by dividing the concentration of the odourants by its nasal odour threshold in
sunflower oil.
I 50 W. GROSCH

Table 6 . Similarity of the odour model mixtures to that of butter sample B"
Odourant present in the model"
Diacetyl &Decalactone Butanoic acid Odour quality' Similarity'
Modcl no. (0.34 mglkg) (4.9 mg/kg) (3.6 mg/kg)

1 - + + Slightly rancid, butanoic acid-like 1

2 i- - + Buttery, slightly rancid 1-2
3 + + - Sweet, buttery 2
4 + + +~ ~ ~~~~~
Typically butter-like, sweet 3
"Amounts of the odourants found in the butter sample B were dissolved in sunflower oil.
*Odour quality of the model.
' Similarity to the odour of the butter sample B (cf. Table 3 ) : 1, weak; 2, medium; 3, strong.

binations (Table 6) and in concentrations equal to Emmental, were added to the unripened cheese
those in B. Actually, the overall odour impression of the mozzarella type. Sensory evaluations indi-
of the model containing the three odourants (no. cated a strong similarity of the odour of the model
4 in Tablc 6) agreed very well with that of sample with that of Emmental. Other odourants showing
B. However, when diacetyl (No. 1) or 6- OAVs between 3 and 14 (Table 7) had no effect
decalactone (no. 2) were lacking the odour inten- on the overall odour of the model; their odours
sity of the models was weaker and a rancid by- were masked in particular by those of the volatile
note was perceivable. acids.
l h e difference in the odour qualities and in the
intensities of the models nos 3 and 4 indicates that
the addition of butanoic acid yielding an OAV of MEAT
76 was necessary to complete the butter-like,
sweet odour profile of sample B. The flavour of heated meat depends strongly on
the animal species and the mode of processing,'3
for example, the odour quality is different when
SWISS CHEESE beef is boiled, roasted or stewed. The last pro-
cess yields the 28 odourants listed in Table 8 . The
AEDA of the volatile fractions obtained from highest OAV of the 11 compounds quantified was
two samples of the Swiss cheese, Emmental, after calculated for 12-methyltridecanal (Table 8). As
separation from the acids resulted in the identi- it was from beef alone, but not from pork,
fication of the compounds listed in Table 7. chicken and turkey, that higher amounts of this
In comparison to the FD-factors, the OAVs of branched aldehyde were liberated during a longer
methional (no. 6 in Table 7) and of the caramel- heating period (Table 9), it was c ~ n c l u d e dthat'~
like smelling furanones nos 9 and 10 were unex- this aldehyde, smelling tallowy and beef-like, is a
pectedly high (Table 7). These differences indi- key compound of the species-specific flavour of
cated that great amountsof nos6,9 and 10 had been beef meat.
lost during the scparation of the acids from the In agreement with this conclusion, 12-methyl-
original extract and during the distillation of the tridecanal was also identified as characteristic
volatiles from the fat dissolved in the extract. odourant of a bouillon prepared from beef. l 4 In
These losses, which affected the results of the first study on the composition of this bouillon
AEDA, were compensated for in the quantitative flavour," the aldehyde had been overlooked,
determination of the odourants by the addition of as its yield was too small in the volatile fraction
internal standards labelled with stable isotopes. I ' obtained by simultaneous distillation and extrac-
The importance of the furanones nos 9 and 10, tion.
which had not been detected bcfore as com- The aldehyde was lacking whcn beef was
ponents of Swiss cheese,"' and of mcthional was roasted for the short period of 7 min." In this
confirmed in a model experiment,I2 in which case, on the basis of their high OAVs ('Table lo),
these odourants, acetic acid, propionic acid, and 2-acetyl-2-thiazoline, 2,3-diethyl-5-methylpyra-
lactic acid, all in the concentrations found in zine and 4-hydroxy-2,5-dimethyl-3(2H)-furanone
 AROMA EXTRACT DILUTION ANALYSIS 151

Table 7. Odourants in the neutral volatile fraction of Emmental cheese“’.”

Sample A Sample B
No. Compound n‘ (Fglkg) OAVh nu (pg/kg) OAV’
1 Butane-2,3-dione 2 n.a. - <2 98 10
2 3-Methylbutanal 2 64 6 4 87 8
3 Ethyl butanoate 7 26 8 7 50 14
4 Ethyl 3-methylbutanoate 4 0.9 9 5 <O.l <I
5 Heptan-2-one 5 380 <I 2 363 <I
6 Methional 5 33 660 5 15 306
7 Oct-I-en-3-one 6 0.5 2 2 0.23 <I
8 Ethyl hexanoate 4 62 3 5 142 7
9 4-Hydroxy-2,5-dimethyl-3(2H)-furanone 7 2035 509 7 824 206
10 5-Ethyl-4-hydroxy-2-methyl-3(2H)-furanone 6 141 94 8 222 148
11 2-sec-Butyl-3methoxypyrazine 6 0.5 2 7 1 .o 4
12 Skatole 8 40 <I 6 26 <I
13 S-Decalactone 9 927 <I 9 1332 <I
Further odourants identified on the basis of AEDA: nonan-2-one, methyl octanoate, (Z)-non-2-enal, (E)-non-
2-enal, ethyl octanoate, vanillin, (Z)-dodec-6-eno-ylactone, S-dodecalactone.
“,bSeefoonotes ‘a’and ‘6’in Table 2.
n.a. = not analysed.

Table 8. Potcnt odourants of stewed

No. Compound nb (mg/kg)‘ OAV”

1 Hexanal 2 0.5 47
2 2-Furfurylthiol 4 0.0015 12
3 Acetic acid 3 172 2.9
4 Methional 5 0.039 22
5 Butyric acid 6 3.33 3.3
6 2-/3-Methylbutyric acid 3 0.18 1.8
7 2-Acetyl-2-thiazoline 4 0.0038 3.8
8 (E,E)-Deca-2,4-dienal 3 0.025 125
9 12-Methyltridecanal 7 0.43 4310
10 4-Hydroxy-2,5-dimethyl-3(2H)-furanone 10 11.66 117
11 4-Hydroxy-4,5-dimethyl-2(5H)-furanone 5 0.013 43
Further odourants identified on the basis of A E D A ( n b s 2): butane-2,3-dione, octanal,
oct-l-en-3-one, 2-acetyl-l-pyrroline, 3-mcrcaptopentan-2-one, nonanal, 2-ethyl-3.5-
dimethylpyrazine, 2.3-diethyl-5-methylpyrazine. decanal, (Z)-non-2-enal, (E)-non-2-enal,
phenylacetaldehyde, (E,E)-nona-2,4-dienaI, rrans-4,5-epoxy-(E)-dec-2-enal, bis(2-methyl-
3-furvl)-disulphide, 4-vinvlauaiacol, y-dodecalactone.
“Beef was roasted in a frying pan for 10 min and then stewed for 4 h in an oven at a
temperature of 200°C. ’’
b E x p n e n t of the FD-factor 2“.
‘ Values of lcvcl based on stewed meat.
”Odour activity values (OAVs) were calculated by dividing the concentrations of the
odourants by their nasal odour thresholds in water.

were mainly responsible for the roasty, earthy BREAD

and caramel-like notes in the odour profile.
The latter furanone, which originates from glu-
cose, fructose and their phosphates,” belongs to
the odourants that steadily increase during heat-
ing of meat. l 4 After 7min of roasting it amounted
to 0.9mg/kg (Table 10) and after 4 h of stewing to
12mg/kg (Table 8).
The pleasant aromaof wheat bread (baguette-type)
depends on the formation of roasty smelling com-
pounds in the baking process and their stability
during storage. Two N-heterocycles, 2-acetyl-l-
pyrroline (no. 6 in Table 11) and 2-acetyltetra-
hydropyridine (no. 7), have been proposed2’**’as
1.52 W.GROSCH

Table 9. Formation of 12-methyltridecanal (I) during boiling alkyl side chain by one carbon yielded with 2-
of the lipids of various animal species at pH 5.7 for 4 h’’ butanoyl-1-pyrroline a compound without the
Lipid source Amount of I (pglkg lipid) roasty smell and with an order of ma nitude higher
odour threshold than that of no. 6 .5-9
Beef (bull), top round 44 Pyrroline no. 6 disappeared very rapidly after
Beef (ox), top round 63 baking.2RThis loss and the retention of odourants
Pork, shoulder 1.6
Lamb, shoulder 14
formed by lipid peroxidation, e.g. (E)-non-2-
Chicken, leg 0.3 enal, resulted in the stale off-odour perceivable
Turkey, fillet 1.6 after storage of wheat bread.’*
As shown in Table 12, rye bread crust in con-
trast to wheat bread was very low in no. 6 and
Table 10. Concentration and OAVs of important odourants much higher in 3-methylbutanal (no. l), methio-
in roasted beef“’” nal (no. 4) and the furanone (no. 18). Further-
more, the results of AEDA (Table 11) suggest
Compound (pglkg)h OAV‘ that also the concentrations of sotolon, the key
4-Hydroxy-2,5-dimethyl-3(2H)-furanone 928 9 odourant of seasonings prepared from plant pro-
2-Acetyl-2-thiazoline 28 28 tein hydroly~ates,~~as well as those of the furanone
2-Ethyl-3,5-dimethylpyrazine 5.4 3 no. 21 and o f acetic acid were significantly higher
2.3-Diethyl-5-methylpyrazine 27 27 in the rye bread crust. All thesc differences in the
Guaiacol 2.I 1
Methional 12.5 7 concentrations o f potent odourants are most likely
the cause for the characteristic diffcrences in the
“Beef was roasted for 7 min in a stainless-steel frying pan overall odours of wheat and rye bread crusts.
containing hydrogenated peanut oil (20 kg per kg beef). Heating of foods accelerates the reactions of
hValucs of level based on raw meat.
‘See footnote ‘c’ in Table 8. monosaccharides with amino acids (Maillard
reaction) and the caramelization of monosaccha-
rides, both with formation of hundreds of volatile
character impact odourants of the roasty note. Of ~ornpounds”.~’of which, however, only a small
these compounds, no. 6 showed the highest FD- fraction may play an active role as food
factor of the odourants identified in the crust, odourants.
whereas the FD-factor of compound no. 7 was As summarized in Table 13, until now the
relatively low. However, during GC analysis of odour potency of a total of 15 compounds has
bread extracts, no. 7 in contrast to no. 6 , was not been detected as being so strong that they have to
stable on several stationary phases. 27 Therefore, be considered as character impact odourants for
to get accurate data, both N-heterocycles and malty, buttery, roasty, earthy and caramel-like
some other important odourants screened by notes in the odour profiles of roasted coffee,
AEDA were quantified in wheat bread crusts by heated meat, bread crusts, roasted sesame seeds
stable isotope dilution assays. According to the and popcorn.
results in Table 12, the level of the acetyltetra-
hydropyridine no. 7 amounts to 94% of that of
the acetylpyrroline no. 6 . Since the odour threshold BEER
of no. 7 in air was three times higher than that of
no. 6,27 the OAV of the former was actually only Wort, the basis of beer, is an aqueous extract of
31% of the latter. These results indicated that no. germinated and roasted cereals. For this reason,
7 belonged to the key odourants of the crust and, it is not surprising that several odourants listed in
in combination with no. 6 , was responsible for the Table 13 (e.g. nos I , 4, 7, 13 and 14) have also
roasty odour note. been detected by AEDA in pale lager beer.36
On the basis of AEDA, nos 6 and 7 contributed 3-Methylbutanol (no. 1 in Table 14) and 2-
also to the roasty odour of popcorn2’ but, in con- phenylethanol (no. 2) are to be stressed, because
trast to wheat bread, 2-furfurylthiol and a homolo- they increased so strongly during fermentation of
gous compound of no. 6, 2-propionyl-l-pyrroline, the wort that, in spite of their relatively high
were additionally involved (Table 13). The odour odour thresholds of l m g / l (water), they, in ad-
threshold of the latter compound was as low as dition to the esters nos 7 and 8, belonged to the
that of no. 6,” but a further elongation of the most important odourants of the different types
 AROMA EXTRACT DILUTION ANALYSIS 153

Table 1 I . A E D A of the crust volatiles from wheat (I) and rye bread (11)22 ''
No. Compound nu in I nu in I1
1 3-Methylbutanal 7 8
2 Diacetyl 6 4
3 (Z)-Hept-4-enal 5 3
4 Methional 6 9
5 Oct-I -en-3-one 5 7
6 2-Acetyl-l -pyrroline 9 <3
7 2-Acetyltetrahydropyridine' (3 n.d.
8 Phen ylacetaldehyde 5 6
9 2-Ethyl-3.5-dimethylpyrazine 4 6
10 Guaiacol <3 5
11 (Z)-Octa- 1,5-dien-3-one 5 4
12 (E)-P-Damascenone 4 7
13 4-Vinyl-2-me thoxyphenol <3 5
14 (Z)-Non-2-enal 6 6
15 (E)-Non-2-enal 8 8
16 (E,E)-Nona-2,4-dienal 4 6
17 (E,E)-Dcca-2,4-dienal 5 9
18 4-Hydroxy-2,5-dimethyl-3(2H)-furanone 7 8
19 3-Hydroxy-2-methylpyran-4-one(maltol) 5 5
20 3-Hydroxy-4,5-dimethyl-2(5H)-furanone(sotolon) <3 5
21 5-Ethyl-3-hydroxy-4-methyl-2(SH)-furanone <3 5
22 Vanillin 6 6
23 Acetic acid 6 9
24 Butanoic acid 7 5
25 2-Methylpropanoic acid 7 4
26 2-/3-Methylbutanoic acid 8 7
27 Phenylacetic acid 5 5

'Exponent of the FD-factor 2".

'Mixture of the tautomeric forms 2-acetyl-l,4,5,6- and 2-acetyl-3,4,5,6-tetrahydropyridine.
n.d. = not detectable.

Table 12. Concentration and O A V s of six odourants in the wheat and rye bread

Crust of
wheat bread rye bread
Compound (pg/kg) OAV" (pglkg) OAV"
~ ~ ~~ ~

3-Methylbutanal (no. 1 in Table 11) 1406 469 3295 1098

Methional (no. 4) 51 510 480 4800
2-Acetyl-I-pyrroline (no. 6) 62' 3116 2.4' 120
2-Acetyltetrahydropyridine(no. 7) 58 976 n.d.
(E)-Non-2-cnal (no. 15) 56 5m 45 450
(no. 18)
4-Hydroxy-2,5-dimethyl-3(2H)-furanone 1920 1920 4310 4310

"Odour activity value (OAV) o n the basis of the odour threshold in air.
'Mean value of two (rye) and three (wheat) different bread samples.
n.d. = not determined.

of beer compared in Table 14. In wheat bread
crumb, in which roasty and caramel-like
odourants were almost absent,27the two alcohols
nos 1 and 2 also originated in the fermentation
process and were responsible for the yeasty odour
note. 37
In pale lager beer the level of the furanone no.
5 was too small to enerate a caramel-like note in
Q
the odour p r ~ f i l e .However,
~ after adjustment of
the concentration of no. 5 found in the dark beer
(Table 14) the caramel-like note was also per-
ceived, in the pale lager.36 An analogous experi-
ment indicated that 4-vinyl-2-methoxyphenol be-
longed to the typical odourants of wheat beer."
154 W. GROSCH

Table 13. Potent odourants formed by the reaction of monosaccharides with amino acids (Maillard reaction) andlor by
caramelization reactions of monosaccharides
Detected by A E D A in:
Bread crust Meat
No. Compound Roasted Roasted
coffee Wheat rye stewed roasted sesame Popcorn
[33, 341 123, 241 [25] 1151 [I71 1351 [291
1 3-Methylbutanal + + +
2 Butane-2,3-dionc + + + + +
3 Pentane-2.3-dione + +
4 Methional + + + + +
5 Phen ylacetaldehyde + + + +
6 2-Furfurylthiol + + + +
7 2-Acetyl-1 -pyrroline + + + +
8 2-Propionyl- 1 -pyrroline +
9 2-Acetyltctrahydropyridine + + +
10 2-Acetyl-2-thiazoline + +
11 2-Ethyl-3.5-dimethylpyrazine + + + + + + +
12 2,3-Diethyl-S-methylpyrazine + + + + +
13 4-Hydroxy-2,5-dimethyl-3(2H)-furanonc + + + + + +
14 3-Hydroxy-4,S-dimethyl-2(5ff)-furanone + + + +
15 le
5-Ethyl-3-hydroxy-4-methyl-2(5H)-furanoi + + +
References are given in brackets.

Table 14. Concentration and OAVs of important odourants in three kinds of beer3'

No. Compound Pale lager bcer Dark lager beer Wheat beer
(mgll) OAV" (mg/l) OAV" (mg/l) OAV"
-
1 3-Methylbutanol 49.6 49.6 42.9 42.9 61.1 61.1
2 2-Phenylethanol 17.5 17.5 9.1 9.1 31.6 31.6
3 Butanoic acid 1.17 I .2 2.19 2.2 1.11 1.1
4 3-Mcthylbutanoic acid 0.24 2.4 0.74 7.4 0.52 5.2
5 4-t-lydroxy-2,5-dimethyI-3(2H)-furanone 0.35 3.5 1.32 13.2 0.54 5.4
6 4-Vinyl-2-methoxyphenol 0.52 5.2 0.25 2.5 0.96 9 .6
7 Ethyl hexanoate 0.149 29.8 0.075 15.0 0.19 38.6
8 Ethyl butanoate 0.061 61.0 0.043 43.0 0.088 88.0
"Odour activity value (OAV) on the basis of the nasal odour threshold in water

GREEN TEA space volumes,'8 indicated that the difference in

AEDA revealed 12 potent odourants in green tea
powder.3ROAVs of these compounds were cal-
culated on the basis of their odour thresholds on
cellulose3' which was used as model substance for
the non-volatile constituents of green tea.
According to the OAVs listed in Table 15, 3-
methylnonane-2,4-dione (smelling strawy), ( Z ) -
hex-3-enal (green) and P-(E)-damascenone
(boiled apple-like) were the most important
odourants. The dione was a degradation product
of furan fatty acids occurring in green tea.39
A new dilution technique, in which the most
potent odourants in the air above the food sample
were evaluated by GC-0 of decreasing head-
the odours of green and black tea was mainly due
to a higher concentration of (Z)-hex-3-enal, (2)-
octa-l,5-dien-3-one and the much lower concen-
tration of linalol in the air above the former.38
DILL HERB (Anethum graveofens L.)
(3R, 4S, 8S)-3,9-Epoxy-l-p-menthene (dill ether),
myristicin. methyl 2-methylbutanoate and (S)-ct-
phellandrcne were screened by AEDA and by
calculation of OAVs as potent odourants of dill
herb.'" In a subsequent sensory study4' (S)-a-
phellandrene was evaluated as the character-
impact compound of the dill flavour which was
 AROMA EXTRACT DILUTION ANALYSIS I55

Table 15. Potent odourants of green tea powderz”-”

No. Compound nu (pg/kg) OAV”

1 (Z)-Octa-l S-dien-3-one 9 1.8 36
2 3-Hydroxy-4.5-dimcthyl-2(5H)-furanone 9 49 38
3 3-Met hylnonane-2,4-dione 9 83 55
4 (Z)-Hcpt-4-enal 8 112 37
5 (Z)-Hex-3-enal 7 101 51
6 (E,Z)-Nona-2,6-dienal 7 61 14
7 Oct-1 -en-3-one 7 6 6
8 (E,E)-Deca-2,4-dienal 7 127 17
9 (E)-P-Damascenone 7 9 45
10 4-Hydroxy-2,5-dimethyl-3(2ff)-furanone 6 276 4
11 243-Methylbutyric acid 6 5280 21
12 2-Phen ylethanol 5 1140 9

“Exponent of the FD-factor 2“.

bodour activity value (OAV) on the basis of the nasal odour threshold of thc
compound on cellulose. ’’

rounded off by an additive effect of the dill ether. lation of OAVs confirmed the changes in the
The contribution of the other two odourants potent odourants during storage of lemon oil. 43.44
appeared less significant. Soybean oil is not stable at room temperature
and under normal atmospheric conditions. A
beany, grassy, hay-like odour and taste, the so-
STUDIES ON OFF-FLAVOURS called reversion flavour, develops when the con-
centration of the peroxides is still low. Storage of
Characterization of off-flavours is a further appli- soybean oil in daylight accelerates the formation
cation of AEDA. Increased levels of odourants of the reversion flavour.
that negatively affect the flavour of a food, the The volatile fractions obtained from soybean
loss of odourants that have a positive impact on oil samples stored in darkness and in daylight
flavours, and the appearance of new odourants were subjected to AEDA.45In this experiment 3-
with off-flavours are all revealed by comparative methylnonane-2,4-dione was screened as
AEDA of fresh and deteriorated samples. The odourant with the highest FD-factor. Actually,
application of this concept is here exemplified by this dione, which in comparison to other decane-
studies on off-flavours formed in lemon oil,42-44 diones had the lowest odour threshold,48
soybean 0 i 1 4 s 3 4 6 and boiled increased strongly when soybean oil was stored in
Lemon oil is an unstable food flavouring, which daylight and it was the most important odourant
deteriorates in the presence of acids and by autoxi- on the basis of its high OAV (Table 17).
dation reactions. As summarized in Table 16, the During storage of soybean oil 3-methylnonane-
monoterpenes neral, geranial and linalol were 2,4-dione was formed by a reaction of singlet
mainly responsible for the odour of the fresh oxygen with furan fatty acids which were iden-
lemon oil. During storage, either under acid con- tified as 10,13-epoxy-l1,12-dimethyloctadeca-
ditions or in the presence of light and oxygen, 10,12-dienoic acid and 12,15-epoxy-13,14-
neral and geranial were completely lost; linalol dimethyleicosa-12,14-dienoicacid.4’
showed a greater stability, in particular in the acid A comparison of Tables 15 and 17 reveals that,
media (Table 16). p-Cresol and p-methyl- in addition to the dione, the green-smelling
acetophenone were detected as potent off- odourants (Z)-hex-3-enal and (Z)-octa-1 S-dien-
flavour substances in both reaction systems. 3-one belong to the most potent odourants of
Differences were mainly due to the formation of green tea and among those causing the reversion
fenchyl alcohol and 1-terpinen-4-01 under acid flavour. This agreement reflects that indeed simi-
conditions as well as carvone and the hydroper- lar green, hay-like odour notes predominate in
oxides nos 18 to 22 in the presence of oxygen and both flavour profiles.
light (Table 16). Quantification of the major The mild flavour of boiled trout (Sufmofurio)is
odourants in both reaction systems and the calcu- affected by the storage of the raw material, for
I56 W. GROSCH

Table 16. AEDA of fresh and stored lemon 0i142.43

Lemon oil
(n in the FD-factor 2")
stored in the
presence of
No. Compound fresh citric acid" oxygen*
1 Myrccne 3 3 n.d.
2 Limonene 3 3 0
3 y-Terpinene 2 1 n.d.
4 Octanal 1 1 0
5 Nonanal 2 1 n.d.
6 Linalol 5 4 1
7 Citronella1 1 n.d. n.d.
8 (Q-Non-2-enal 2 1 2
9 Decanal 2 2 2
10 Neral 6 n.d. n.d.
11 Geranial 6 n.d. n.d.
12 p-Cymene n.d. 1 2
13 Fenchyl alcohol n.d. 1 n.d.
14 I-Terpinen-4-01 n.d. 1 n.d.
15 p-Cresol n.d. 2 3
16 p-Methylacetophenone n.d. 4 5
17 Carvone n.d. n.d. 6
18 4-Acetyl-l-methylcyclohex-l-ene n.d. n.d. 3
19 cis-p-Mentha-2,8-dien-l-hydroperoxide n.d. n.d. 3
20 Irons-p-Mentha-[ 1(7).8]-dien-2-hydroproxide n.d. n.d. 3
21 p-Mentha-l,8-dien-3-hydroperoxide n.d. n.d. 3
22 trans-p-Men tha-6.8-dien-hydroperoxide n.d. n.d. 3
"Lemon oil was emulsified in 5% (wlv) aqueous citric acid solution and stored for 30 days
at 37°C.
the presence of oxygen lemon oil was irradiated for 120 h.
n.d. = not detectable.

Table 17. Concentrations and OAVs of odourants in a sample of soybean oil

stored for 30 days at room temwrature in darkness or in davlieht4"
~~

Storage
in darkness in daylight
No. Compound (Fglkg) OAV" (Fglkg) OAV"
-
1 Hexenal 54 <I 2367 32
2 (Z)-Hex-3-enal 1.3 <1 94 34
3 Oct - I -en-3-one 0.3 1 11 36
4 (Z)-Octa-l,5-dien-3-one C0.05 0.6 20
5 (E)-Non-2-enal 4.1 c1 28 <I
6 (Z)-Non-2-enal 0.2 <1 4.4 7.3
7 trans-4,5-Epoxy-( E)-dec-2-enal n.d. 12 4
8 3-Methylnonane-2.4-dione 0.9 c1 721 481
"On the basis of the retronasal odour threshold in sunflower oil.
n.d. = not detectable.

example, after 17 weeks at -13°C they exhibited
a fatty, fishy ~ d o u r . ~Calculation
' of OAVs re-
vealed (Z)-octa-l,5-dien-3-one,(E,Z)-nona-2,6-
dienal and methional as the most potent
odourants of the freshly boiled trout (Table 18).
The OAVs increased when the raw trout were
stored as mentioned above. In particular, the
OAV of (Z)-hex-3-enal and (Z, Z)-nona-3,6-
 AROMA EXTRACT DILUTION ANALYSIS 157

Tablc 18. Concentration and OAVs of potent odourants of boiled trout before and after storage
of the raw material4’
Freshly boiled Boiled after storage“
No. Compound (Wkg) OAVb (CLg/kg) OAVb
1 Butane-2.3-dione 268 54 363 73
2 Pentane-2,3dione 140 28 167 33
3 (Z)-Hex-3-enal 1.6 53 13 430
4 Hexanal 37 4 98 9
5 (Z)-HeptA-enal 2.8 46 6.0 100
6 Methional 5.8 145 7.5 188
7 Oct-1 -en-3-one 0.6 60 0.9 90
8 (Z)-Octa-1,5-dien-3-one 0.36 900 0.64 1600
9 (Z,Z)-Nona-3,6-dienaI <1.2 <24 14.0 280
10 (E)-Non-2-enal 2.7 34 5.0 62
11 (E,Z)-Nona-2,6-dienal 8.0 400 10.5 525
12 (E,E)-Nona-2,4-dienaI 4.7 78 6.4 107
“The trout were stored for 17 weeks at -13°C before boiling.
’OAV on the basis of the nasal odour threshold in water.

dienal increased so strongly that these aldehydes, hibited the lowest and hexanal,” (E)-non-2-
in addition to (Z)-octa-l,5-dien-3-one and ( E ,2)- enal6 and (E,E)-de~a-2,4-dienaI~~ the highest
nona-2,6-dienal, showed the highest OAVs. odour thresholds. Nevertheless, the latter three
The study on boiled trout, as well as those on may belong to the potent odourants of a food when
olive oil (Table 2), butter (Table 4), green tea they occur in relatively high concentration levels.
(Table 15) and soybean oil (Table 17), indicates
that only a small number of the numerous volatiles
formed by the peroxidation of polyunsaturated CONCLUSION
fatted acids [review in Reference 501 are
odourants so potent that they can play a role in During the last decade, knowledge about the
flavours and off-flavours. Among these components causing the characteristic odour
odourants, which are summarized in relation to notes of heated and of fatty foods in particular has
their precursors in Table 19, (Z)-octa-l,5-dien-3- been significantly increased by AEDA, quanti-
one” and trans-4,5-epoxy-(E)-de~-2-enal~’*~~ ex- fication of odourants showing high FD-factors
and calculation of OAVs. However, AEDA is
limited to odourants with boiling points higher
Table 19. Potent odourants formed by the peroxidation of than that of the solvent used for extraction of the
PUFA 50 food. Hence, as recently the identi-
Odourant fication experiments have to be completed by GC-
0 of decreasing headspace volumes, which reveals
A . n-6 PUFA, e.g. linoleic acid the potent highly volatile odour compounds.
Hexanal As discussed in the review, analytical pro-
Oct-1-en-3-one
(E)-Non-2-enal cedures have been developed for the accurate
(Z)-Non-2-enal determination of labile odourants and/or for
(E,E)-Deca-2,4-dienal those occurring in traces. The methods can help
(E,Z)-Deca-2,4-dienal to measure such compounds in foods, for example,
trans-4,5-Epoxy-( Q-dec-2-enal
in their dependence on the ingredients, on the
B. n-3 PUFA, e.g. linolenic acid parameters of processing and storage conditions,
(Z)-Hex-3-enal and also in model systems.
(Z)-Hept-4-enal
(Z)-Oct-l,5-dien-3-one
(E,Z)-Nona-2,6-dienal Acknowledgements-I wish to thank Professor Dr P.Schieber-
(Z,Z)-Nona-3,6-dienaI le, University Wuppertal, for some articles in press, M n Dr
A. Sen, Munich, for proof reading and Mrs R. Berger for
“PUFA = polyunsaturated fatty acid. typing the manuscript.
I58 W. GROSCH
REFERENCES
I . T. E. Acree in Flavour Science. Sensible Principles and
Techniques. ed. T. E. Acrcc and R. Teranishi. p. I , ACS
Professional Reference Book, American Chemical Society,
Washington (1993).
2. W. Grosch, Trends Food Sci. Technol.. 4, 68 (1993).
3. N. Abbott, P. Etievant, S. lssanchou and D. Langois, J .
Agric. Food Chem.. 41. 1698 (1993).
4. F. Ullrich and W. Grosch, Z . Lebens. Unters. Forsch.,
184. 277 (1987).
5. H. Guth and W. Grosch, Fefte Seifen Ansfrichm., 93. 335
(1991 ).
6. H. Guth and W. Grosch, 1. A m . Oil Chem. Soc.. 70. 513
(1993).
7. B. Blekas, H. Guth and W. Grosch, in Trends in Flavour
Research, cd. H. Maarse and D. G. Van der Hcij, p. 499,
Elsevier, Amsterdam (1994).
8. P. Schieberle. K . Gassenmeier, H. Guth, A. Sen and W.
Grosch, Lebensm. W i n . Technol.. 26, 347 (1993).
9. S. Widder, A. Sen and W. Grosch, Z. Lebensm. Unfers.
Forsch.. 193, 32 (1991).
10. M. Prciningcr, M. Rychlik and W. Grosch in Trends in
Flavour Research, ed. H. Maarsc and D. G. Van der Heij.
p. 267. Elsevier, Amsterdam (1994).
11. M. Preiningcr and W. Grosch, 1.ehensrn. Wiss. Technol
(in press).
12. M. Preininger, R. Warmke, H.-D. Belitz and W. Grosch
in Procecdings of the 4th Wartburg Aroma Symposium,
Eiscnach, 1-4 March 1994 (submittcd).
13. G . Mac1.eod and B. M. Coppock, 1. Food Sci., 43, 145
(1978).
14. W. Grosch, G . Zciler-Hilgart, C. Cerny and H. Guth in
Progress in Flavour Precursor Studies. ed. P. Schreier and
P. Winterhalter, p. 329, Allured Publishing Corporation,
Carol Stream (1993).
15. H. Guth and W. Grosch, Lebensm. Wiss. Technol.. 26,
171 (1993).
16. U. Gasser and W. Grosch, Z . Lehensm. Unfers. Forsch.,
186, 489 (1988).
17. C. Cerny and W. Grosch, Z . Lebensm. Unters. Forsch.,
194, 322 (1992).
18. C . Ccrny and W. Grosch, Z . 1,ebensm. Unfers. Forsch.,
196, 417 (1993).
19. P. Schicbcrle in Flavour Precursors. ed. R. Teranishi, G .
R. Takcoka and M. Giintert, p. 164, ACS Symposium
Series 490. American Chemical Society, Washington,
DC (1992).
20. P. Schicberle and W. Grosch, Z. Lebensm. Unfers.
Forsch., 180, 474 (1985).
21. I. R. Hunter, M. K . Walden, J. R. Scherer and R. E.
Lundin, Cereal Chem., 46, 189 (1969).
22. P. Schicbcrlc and W. Grosch, Z . 1,ebensm. llnfers.
Forsch., 185, 1 1 1 (1987).
23. P. Schieberle and W. Grosch in Thermal Generation of
Aromas, ed. T. H . Parliment, R. J. McGorrin and C. T. 49. H. Guth and W. Grosch, Fette Seifen Ansrrichm., 93, 249
Ho, p. 258, ACS Symposium Series 409, American
Chemical Society, Washington. DC (1989).
24. P. Schichcrle in The Maillard Reacrion in Food Process-
ing, Human Nitfrifionand Physiology. ed. P. A. Finot, 51. P. A. T. Swoboda and K . E. Peers, J . Sci. Food Agric.,
H. U . Aeschbachcr, K.F. Hurrcll and K.Liardon, p. 187,
Rirkhauser. Basel (1990).
25. P. Schieberle and W. Grosch, Z . Lebensm. Unters.
Forsch., 198, 292 (1994).
26. P. Schieberlc and W. Grosch, J . Agric. Food Chem., 35,
252 (1987).
27. P. Schicberle and W. Grosch. Z. Lebensm. Unfers.
Forsch., 192, 130 (1991).
28. P. Schieberle and W. Grosch, Flavour Fragrance J . , 7,213
(1992).
29. P. Schiebcrlc, 1. Food Agric. Chern., 39, 1141 (1991).
30. 1. Blank, P. Schieberle and W. Grosch in Progress in
Flavour Precursor Studies, ed. P. Schreier and P. Win-
terhalter, p. 103. Allured Publishing Corporation, Carol
Stream (1993).
31. R. Tressl, B. Helak, N. Martin and E. Kersten in Thermal
Generation of Aromas, ed. T. H. Parliment, R. J.
McGorrin and C.-T. Ho, p. 156, ACS Symposium Series
409, American Chemical Society, Washington, DC
(1989).
32. W. Baltcs in The Maillard Reacrion in Food Processing,
Human Nutrition and Physiology, ed. P. A. Finot, H. U.
Acschbacher, R. F. Hurrell and R. 1-iardon. p. 43, Birk-
hiuscr, Basel (1990).
33. W. Holscher. 0. G . Vitzthum and H. Steinhart, Cafe,
Cacao, The, 34, 205 (1990).
34. I. Blank, A. Sen and W. Grosch, Z . Lebensm. Unfers.
Forsch., 195, 239 (1992).
35. P. Schicberlc in Progress in Flavour Precursor Srudies. ed.
P. Schreier and P. Winterhalter, p. 343, Allured Pub-
lishing Corporation, Carol Stream (1993).
36. P. Schiebcrle, Z . Lebensrn. Unfers. Forsch., 193, 558
( I991 ).
37. K. Gassenmeier and P. Schieberlc, Z . Lebensm. Unters.
Forsch. (in press).
38. H. Guth and W. Grosch, Flavour Fragrance J . , 8. 173
(1993).
39. H. Guth and W. Grosch in Progress in Flavour Precursor
Sfitdies, cd. P. Schreicr and P. Winterhalter, p. 401,
Allured Publishing Corporation, Carol Stream (1993).
40. 1. Blank and W. Grosch. J . Food Sci., 56, 63 (1991).
41. 1. Blank, A. Scn and W. Grosch. Food Chem., 43, 337
( 1992).
42. P. Schicberlc and W. Grosch, J . Agric Food Chem., 36,
797 (1988).
43. P. Schieberle and W. Grosch, Z . Lebensm. Unrers.
Forsch.. 189. 26 (1989).
44. P. Schieberle and W. Grosch. Lebensm. Wiss. Technol..
21, 158 (1988).
45. H. Guth and W. Grosch, Lebensm. Wiss. Technol.. 23. 59
(1990).
46. H. Guth and W. Grosch, Lebensm. Wiss. Technol., 23,
512 (1990).
47. C. Milo and W. Grosch, J . Agric. Food Chern., 41, 2076
(1993).
48. H. Guth and W. Grosch, Ferre Seifen Ansrrichrn., 91, 225
( 1989).
( 1991).
50. W. Grosch in Autoxidafion of Unsaturated Lipids, cd. H.
W . 4 . Chan, p. 95, Academic Press, London (1987).
28. 1019 (1977).
52. P. W. Meijboom, J . A m . Oil Chem. Soc., 41, 326 (1964).