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Springer-Verlag 2000
O R IG IN AL P AP E R
Introduction
The flavour of coffee changes when a brew is prepared from ground roasted coffee. Caramel-like, buttery and phenolic notes become more intense. Screening for potent odorants, the results of which have
273
Chemicals
Pure samples of the odorants listed in Table 1 were purchased
from the sources given in parentheses or synthesized according
to the cited literature: 17, 9, 11, 12, 14, 16, 1921 (Aldrich,
Steinheim, Germany), 8, 10 [5], 13 (Givaudan-Roure, Duebendorf, Switzerland), 15 (Haarmann & Reimer, Holzminden, Germany), 17, 18 (Lancaster, Muehlheim/Ruhr, Germany), 22 [4],
23 [6], 24 [3], 25 (Fluka, Neu-Ulm, Germany). Stable isotopomers of the odorants used as internal standards were obtained as
reported [7]. Sodium dihydrogen phosphate, disodium hydrogen
phoshapate and p-hydroxymercuribenzoic acid were purchased
from Aldrich (Steinheim, Germany), and cysteine was obtained
from Merck (Darmstadt, Germany).
Methods
High-resolution gas chromatography mass spectrometry
Static headspace analysis mass spectrometry (SHA-MS) and
multidimensional gas chromatography mass spectrometry
(MDGC-MS) were carried out according to Mayer et al. [7].
Table 1 Concentrations of the
odorants in coffee powder and
coffee brew, and extraction
yields of the compounds
Compound
Acetaldehyde (1)
Methylpropanal (2)
2-Methylbutanal (3)
3-Methylbutanal (4)
2,3-Butanedione (5)
2,3-Pentanedione (6)
2-Ethyl-3,5-dimethylpyrazine (7)
2-Ethenyl-3,5-dimethylpyrazine (8)
2,3-Diethyl-5-methylpyrazine (9)
2-Ethenyl-3-ethyl-5-methylpyryzine (10)
3-Isobutyl-2-methoxypyrazine (11)
4-Hydroxy-2,5-dimethyl-3(2H)-furanone (12)
2(5)-Ethyl-4-hydroxy-5(2)-methyl-3(2H)-furanone (13)
3-Hydroxy-4,5-dimethyl-2(5H)-furanone (14)
(E)-b-Damascenone (15)
Guaiacol (16)
4-Ethylguaiacol (17)
4-Vinylguaiacol (18)
Vanillin (19)
2-Furfurylthiol (20)
Methional (21)
3-Mercapto-3-methylbutyl formate (22)
2-Methyl-3-furanthiol (23)
3-Methyl-2-buten-1-thiol (24)
Methanethiol (25)
a
b
Concentrationa
Extraction
Powder
(mg/kg)
Brew
(mg/l)
yieldb
120000
124000
126000
117000
149000
135000
111400
111153
111100
111115
111120
140000
116000
111900
111260
112400
111800
145000
114100
111700
111250
111130
111160
111113
114400
4700
1760
1870
1570
2100
1600
1117
1111.0
1113.6
1110.2
1111.5
7200
1800
1180
1111.6
1120
1148
1740
1210
1117
1110
1115.7
1111.1
1110.6
1170
73
59
62
62
79
85
79
35
67
25
23
95
93
78
11
65
49
30
95
19
74
81
34
85
72
Data are means of at least two assays that did not vary by more than 10 %
Percentage ratio of the odorant amount in 1 l coffee brew and in 54 g coffee powder
274
microdistillation [9]. An aliquot was then analysed by MDGCMS.
Odorants 23 and 24. Coffee brew (500 ml) was spiked with
known amounts of the labelled internal standards and extracted
with dichloromethane (2 250 ml). The organic layers were isolated by centrifugation and concentrated to 80 ml by distilling
off the solvent at 40 C under reduced pressure (60 kPa). p-Hydroxymercuribenzoic acid (2.5 mmol/l in disodium hydrogen
phosphate/sodium dihydrogen phosphate buffer, 0.1 mol/l, pH
8.5, 3 25 ml) was added to the extract and stirred at 600 rpm
for 15 min to trap all of the thiols as salts [10]. After separation,
the combined aqueous layers were concentrated to 10 ml by distillation under reduced pressure (60 C, 15 kPa). Subsequently,
cysteine as displacing agent was added and the liberated thiols
were trapped by a dynamic headspace technique and analysed
by HRGC-MS according to Kerscher an Grosch [6].
Odorant 25. Coffee brew (50 ml) was put into a vessel (300 ml)
which was sealed with a septum. A known amount of [2H3]methanethiol [11] was injected by a gas-tight syringe. The vessel was
kept at 20 C for 30 min, and an aliquot of the headspace
(10 ml) was analysed by SHA-MS.
Sensory experiments
Sensory analyses were performed as described previously [4].
Models
Stock solutions, in which the concentrations of the odorants
were 1000-fold higher than in coffee brew (Table 1), were prepared as follows: compounds 16, 12 and 13 were dissolved in
water. The following odorants were available in various solvents:
9, 11, 14, 1622 (ethanol), 7, 8 (diethyl ether), 15, 24 (dichloromethane), 23 (pentane). The amount calculated for the stock
solution was diluted with water.
Pyrazine 10 was compensated for by increasing correspondingly the concentration of pyrazine 8 [4]. The stock solution of
25 was prepared by dissolving 7.8 ml of gas in water (100 ml).
To prepare the aroma model, 1 ml of each stock solution was
pipetted into phosphate buffer (disodium hydrogen phosphate/
sodium dihydrogen phosphate, 0.1 mol/l, pH 5.4), the volume of
which was adjusted to 1 l. The solutions (20 ml) were poured
into glass beakers (diameter 40 mm, capacity 45 ml) and
coloured dark brown with odourless sugar colour (0.1 ml; Kattus, Maisach, Germany).
Odour profile
Samples were presented to a test panel consisting of 10 experienced participants: six males and four females aged 2635
years. They scored the intensities of the odour attributes sweetish/caramel-like, earthy, roasty/sulfurous and smoky on a
scale from 0 (absent) to 3 (very strong). Furthermore, the assessors evaluated the similarity of the aroma of the model to that
of the coffee brew on a scale from 0 (no similarity) to 3 (identical with coffee brew). After an outlier test according to Nalimov
[12], the results were expressed as means standard deviations.
Omission experiments
One or more compounds (Table 3) were omitted in the aroma
model of coffee brew. Each reduced model was presented
together with two complete models to 10 assessors who, by sniffing the sample, had to find out what was different in the overall
odour. The triangle test was repeated on the same day. The
number of correct answers was summed up (maximum 20), and
the significance of the result (P < 0.05) was evaluated [13].
Omission experiments
An aroma model for the coffee brew was prepared on
the basis of the quantitative data listed in Table 1.
Ten assessors described its overall odour as clearly
275
Table 2 Odour profiles of the coffee brew and the aroma modela
Attribute
Coffee brew
Aroma model
Sweetish/caramel-like
Earthy
Roasty/sulfurous
Smoky
Similarityc
1.6
1.9
2.3
2.0
2.1
1.7
2.1
1.4
2.0
(0.4)
(0.2)
(0.5)
(0.6)
(0.7)b
(0.4)
(0.6)
(0.4)b
(0.3)
Experiment
no.
Compound(s) omitted
11
12
Acetaldehyde (1)
Acetaldehyde (1), methylpropanal (2), 2-methylbutanal (3),
3-methylbutanal (4)
2,3-Butanedione (5), 2,3-pentanedione (6)
Acetaldehyde (1), methylpropanal (2), 2-methylbutanal (3),
3-methylbutanal (4), 2,3-butanedione (5), 2,3-pentanedione (6)
2-Ethyl-3,5-dimethylpyrazine (7), 2-ethenyl-3,5-dimethylpyrazine (8),
2,3-diethyl-5-methylpyrazine (9), 3-isobutyl-2-methoxypyrazine (11)
2-Ethyl-3,5-dimethylpyrazine (7), 2-ethenyl-3,5-dimethylpyrazine (8),
2,3-diethyl-5-methylpyryzine (9)
2-Ethyl-3,5-dimethylpyrazine (7)
3-Isobutyl-2-methoxypyrazine (11)
4-Hydroxy-2,5-dimethyl-3(2H)-furanone (12), 2(5)-ethyl-4-hydroxy5(2)-methyl-3(2H)-furanone (13), 3-hydroxy-4,5-dimethyl-2(5H)furanone (14)
4-Hydroxy-2,5-dimethyl-3(2H)-furanone (12), 2(5)-ethyl-4-hydroxy5(2)-methyl-3(2H)-furanone (13)
3-Hydroxy-4,5-dimethyl-2(5H)-furanone (14)
(E)-b-Damascenone (15)
Guaiacol (16), 4-ethylguaiacol (17), 4-vinylguaiacol (18), vanillin
(19)
Guaiacol (16)
4-Vinylguaiacol (18)
2-Furfurylthiol (20), methional (21), 3-mercapto-3-methylbutyl formate (22), 2-methyl-3-furanthiol (23), 3-methyl-2-buten-1-thiol (24),
methanethiol (25)
2-Furfurylthiol (20)
Methional (21), 3-mercapto-3-methylbutyl formate (22), 2-methyl3-furanthiol (23), 3-methyl-2-buten-1-thiol (24), methanethiol (25)
Methional (21)
3-Mercapto-3-methylbutyl formate (22)
2-Methyl-3-furanthiol (23)
3-Methyl-2-buten-1-thiol (24)
Methanethiol (25)
Acetaldehyde (1), methanethiol (25)
13
14
15
16
17
18
19
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
a
Number of
assessors b
6
8
4
12c
13c
12c
4
6
12c
7
8
6
12c
7
8
18c
12c
12c
14c
12c
7
6
8
13c
Models lacking in one or more components (numbering refers to Table 1) were singly compared
to two models containing the complete set of 25 odorants
b
Number of correct answers (maximum 20)
c
Significant difference (P < 0.05) in the flavour between the complete and the reduced model [14]
276
The composition of the brew aroma model was varied to identify the character impact flavour compounds. According to the results in Table 3, the omission of acetaldehyde (1) in exp. 1, the Strecker
aldehydes 14 in exp. 2 and the diones 5 and 6 in exp.
3 was not significantly perceived by the sensory panel.
This was in contrast to exp. 4 in which the lack of carbonyl compounds 16 changed the overall odour so
clearly that a significant number of answers were correct. The alkylpyrazines 79 with (exp. 5) and without
(exp. 6) methoxypyrazine 11 were clearly missed by
the assessors, but not pyrazine 7 (exp. 7) and methoxypyrazine 11 (exp. 8) alone. In case of the furanones the absence of furaneol (12) and ethylfuraneol
(13) in exp. 10 as well as of sotolon (14) in exp. 11
was not recognized. Only when the three furanones
were lacking in exp. 9 was a change in the aroma perceived by a significant number of assessors. The finding that single compounds, but not the complete class,
could be omitted without a clear effect on the aroma
was also observed for the phenolic odorants. When
omitting the whole group (exp. 13), the assessors perceived a weakening of the smoky note in the reduced
model. In contrast, guaiacol (16) in exp. 14 or 4-vinylguaiacol (18) in exp. 15 were not clearly missed.
Exp. 16 indicates that the group of sulfur compounds had the greatest effect on the aroma of the
model, as 18 of the answers were correct. The roasty/
sulfurous note was weaker in the reduced model. As
shown in exp. 17, 2-furfurylthiol (20) contributed to
the aroma. However, when it was the only sulfur compound in the reduced model (exp. 18), the odour difference to the complete model was not abolished. This
was mainly due to methional (21) and 3-mercapto3-methylbutyl formate (22) since exps. 19 and 20 indicated that 21 and 22 were further sulfur compounds
whose absence impaired the aroma. In contrast, the
odour contributions of thiols 2325 were weak (exps.
2123). According to exp. 24, the aroma of the
reduced model differed clearly from that of the complete one when methanethiol (25) together with acetaldehyde (1) were lacking.
A comparison of the results presented here with
those obtained for the model imitating the aroma of
ground roasted coffee [4] reveals major differences in
the contributions of the sulfur compounds. The effect
of 2-furfurylthiol (20), the predominating odorant of
the powder, was weaker in the brew because it its
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