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Eur Food Res Technol

DOI 10.1007/s00217-012-1812-x

ORIGINAL PAPER

Analysis and stereodifferentiation of linalool in Theobroma cacao


and cocoa products using enantioselective multidimensional gas
chromatography
Hans-Georg Schmarr Karl-Heinz Engel

Received: 21 June 2012 / Revised: 3 August 2012 / Accepted: 11 August 2012


Springer-Verlag 2012

Abstract Contents and enantiomeric distributions of


linalool (3,7-dimethyl-1,6-octadien-3-ol) were investigated
in raw and roasted cocoa beans (seeds of Theobroma
cacao) of defined origin as well as in commercial products,
such as cocoa powders or chocolates. The stereodifferentiation of linalool was achieved by enantioselective
multidimensional gas chromatography using heptakis
(2,3-di-O-methyl-6-O-tert-butyldimethylsilyl)-b-cyclodextrin as chiral stationary phase. Simultaneous steam distillationextraction at pH 7 allowed sample preparation
without racemization of linalool. Cocoa beans contain
linalool primarily as (S)-enantiomer. Model experiments
and analyses of commercial products such as cocoa powders and chocolates revealed that technological procedures
employed in the manufacturing of cocoa products do not
result in significant changes of the original enantiomeric
distribution of linalool.
Keywords Theobroma cacao  Cocoa  Stereoisomeric
flavour compounds  Linalool  Multidimensional gas
chromatography

This paper is dedicated to Prof. Dr. Armin Mosandl (Dettelbach) on


the occasion of his 70th birthday.
H.-G. Schmarr (&)
Dienstleistungszentrum Landlicher Raum Rheinpfalz,
Kompetenzzentrum Weinforschung, Breitenweg 71,
67435 Neustadt an der Weinstrae, Germany
e-mail: hans-georg.schmarr@dlr.rlp.de
H.-G. Schmarr  K.-H. Engel
Technische Universitat Munchen, Lehrstuhl fur Allgemeine
Lebensmitteltechnologie, Maximus-von-Imhof-Forum 2,
85350 Freising-Weihenstephan, Germany

Introduction
The flavour of cocoa beans (seeds of Theobroma cacao)
and products thereof is not only dependent on the genotype
but largely determined by chemical and microbial reactions
during processing and manufacturing. Fermentation and
drying of the raw cocoa beans are the first essential steps
for the development of the characteristic flavour. Reactions
occurring during fermentation depend on the water content
and are catalysed by enzymes of the living seed; they are
followed by non-enzymatic reactions in a dry lipid phase.
The resulting dried cocoa beans are further treated to
produce various cocoa products and chocolates. Roasting
of the raw cocoa beans is the major step during this manufacturing and essential for the aroma of the final product.
Comprehensive overviews on cocoa processing have been
published [1, 2].
Since the first report on the study of cocoa aroma in
1912, in which linalool was identified amongst the main
volatile constituents in an extract prepared from about 2
tons of roasted cocoa beans [3], hundreds of volatile
compounds have been identified in different types of cocoa
[4]. In recent years, studies focused on the investigation of
the aroma-relevant compounds in cocoa and chocolate
[57] and their changes during the roasting process [8].
Within the terpene fraction of the cocoa aroma, linalool
(3,7-dimethyl-1,6-octadien-3-ol) is an important component and contributes to the flowery and tea-like flavour of
some cocoa varieties, for example, the Criollo type [9, 10].
Such most priced Criollo cocoas are classified as flavourgrade because of their aromatic, mild, flowery, and tea-like flavour quality. They are mainly harvested in Venezuela,
Trinidad, and Ecuador (Arriba) and contain significant
amounts of linalool (0.52 mg kg-1) and other terpenoids,
which have been reported to contribute to their valuable

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Eur Food Res Technol

impression [2]. On the other hand, basic cocoa varieties, such


as the Forastero type, exhibit relatively low concentrations of
linalool. The linalool content as such and the concentration
ratio of linalool versus benzaldehyde have been described as
markers for the industrial quality control of unroasted cocoa
beans [9, 11]. Despite the increase in knowledge on aroma
compounds present in cocoa products today, the determination of the ratio of linalool and benzaldehyde remains a
pragmatic parameter, still suitable for industrial quality
control.
The enantiomeric composition of linalool in cocoa
varieties or cocoa products has, to the authors knowledge,
not yet been investigated.
In general, the enantioselective analysis of chiral aroma
compounds has been widely used in the quality and
authenticity control of a large variety of flavours and
essential oils [1216]. It is also well known that the sensory
properties of enantiomers may vary significantly [17, 18].
In the case of linalool, the olfactory characteristics have
been described as sweet, petit grain for the (S)-enantiomer
(coriandrol), and as lavender, woody for the (R)-enantiomer (licareol) [19]. The odour threshold in water for (R)linalool (0.8 lg/L-1) is significantly lower than that of
(S)-linalool (7.4 lg/L-1) [20]. The sensory potency of the
(R)-enantiomer has been confirmed by other evaluations
[2123]. For racemic linalool, an orthonasal odour threshold
of 37 lg/kg-1 was determined in sunflower oil [6].
Most of the studies published on the enantiomeric distribution of linalool in various essential oils and fruits used
hyphenated techniques for the enantioselective analysis.
Online-coupled high-performance liquid chromatography
with gas chromatography (HPLC-GC) [24] or multidimensional gas chromatography (MDGC) is often employed
as analytical techniques [25, 26]. Techniques and applications of such multidimensional chromatographic approaches have recently been reviewed [27]. For linalool, it was
shown that the naturally occurring enantiomeric compositions can vary significantly. Essential oils of Lavandula
species primarily contain (R)-linalool, whereas in essential
oils of Melissa officinalis L., linalool is found as an almost
racemic mixture [15]. Interestingly, in the case of sweet and
bitter orange oils, different enantiomeric ratios were found.
Whilst (S)-linalool predominates in sweet orange oils, the
opposite enantiomer is present in bitter orange oils [24].
Results published for tea (Camellia sinensis Var.) showed
no clear preference for an enantiomer, which is probably
due to the many enzymatic processes involved during fermentation of tea leaves [28, 29]. A clear preponderance of
the (S)-enantiomer of linalool found in Morio-Muscat grape
musts changed to racemic linalool after fermentation and
bottle maturation of the final wine [30].
Furthermore, it has also been shown that the methods
used for sample preparation prior to enantioselective

123

analysis play an important role in the determination of the


enantiomeric distribution of linalool. This is due to the
instability of linalool in acidic medium, a well-known fact
[31, 32] that may result in partial racemization [33].
However, using neutral pH conditions, no racemization
was found under extractive or simultaneous steam distillationextraction (SDE) conditions [25].
Due to the practical importance of linalool for the flavour and quality control of cocoa beans, the objective of
this study was to investigate the linalool content, its concentration ratio versus benzaldehyde as well as the enantiomeric distribution of linalool in various cocoa beans of
different origin. Furthermore, the potential influence of the
roasting process on the original enantiomeric distribution
of linalool should be studied.

Materials and methods


Unroasted (raw) and roasted cocoa beans of defined origin
were provided by the Fraunhofer Institut fur Lebensmitteltechnologie und Verpackung (Freising, Germany), the
Bundesverband der Deutschen Suwarenindustrie e.V.
(Koln, Germany). HACHEZ (Bremen, Germany), Kraft
Jacobs Suchard R&D, Inc. (Munchen, Germany), as well as
the Verein der am Rohkakaohandel beteiligten Firmen
e.V. (Hamburg, Germany). Commercial cocoa products
were purchased in groceries in Germany and Switzerland.
Cocoa beans were peeled (separation of the nibs) and
thereafter milled in a coffee mill with addition of liquid
nitrogen to form a fine powder. Commercial powder was
used as such. Chocolate was deep-frozen at minus 70 C
and then milled as described for the cocoa beans. One
sample of cocoa beans (Arriba, about 100 g) was roasted in
the laboratory under the following conditions: The peeled
and unroasted beans were milled with liquid nitrogen to a
particle size of about 2 mm. The pieces were then spread
on a baking sheet covered with aluminium foil to form a
thin layer. Roasting was performed for 30 min in an oven
heated to 140 C. Such conditions are in the upper temperature, and time range normally applied in industrial
roasting processes [34]. After roasting, the powder was
further milled to a fine powder as described above. Solvents (re-distilled before use) and chemicals were obtained
from Merck (Darmstadt, Germany), racemic linalool and
(R)-(-)-linalool were from Fluka (Buchs, Switzerland).
Liquid extraction
Ground cocoa beans (50 g) were suspended in 500 mL of a
phosphate buffer (322 mL 0.5 m Na2HPO4 ? 178 mL
0.5 m NaH2PO4; pH = 7 0.2) and continuously extracted for 24 h with 120 mL n-pentane ? dichloromethane

Eur Food Res Technol

(1 ? 1; v ? v) using a perforation apparatus according to


Kutscher-Steudel [35]. The resulting extract was dried with
anhydrous sodium sulphate and concentrated using a Vigreux column. Fat could be separated by storing the extract
at ?4 C and using the supernatant extract for further gas
chromatographic analysis.
Simultaneous steam distillationextraction (SDE)
Ground cocoa beans, commercial powder, or ground chocolate (50 g) was suspended in 500 mL of a phosphate buffer
solution of pH 7 (as described before) and brought to a volume
of about 1 L with deionised water. Then, 0.1 mg (100 lL of
a 1 mg mL-1 solution in diethyl ether) of 1-octanol was
added as an internal standard, and the suspension was distilled
and continuously extracted for 1 h with 120 mL diethyl
ether ? n-pentane (1 ? 1; v ? v) in a LikensNickerson
apparatus, modified according to Schulz et al. [36]. The
extract obtained was dried over anhydrous sodium sulphate
and concentrated to about 1 mL using a Vigreux column. The
SDE approach resulted in complete recovery ([99 %) of
linalool (4 mg) from an aqueous solution (pH 7) after 1 h.
Racemization test for linalool
Liquidliquid extraction
Enantiomerically pure (R)-linalool (100 lL of a 1 mg L-1
solution) was added to aqueous solutions of different pH
values: pH 3.5 0.2 (citrate buffer), 7.0 0.2 (phosphate
buffer), and 9.0 0.2 (glycine buffer); buffers were prepared according to standard procedures (Labor-Tools,
Merck, Darmstadt, Germany). These solutions were
refluxed for 1 h, extracted with diethyl ether, dried with
anhydrous sodium sulphate and concentrated to about
1 mL using a Vigreux column.
SDE
Enantiomerically pure (R)-linalool (100 lL of a 1 mg L-1
solution) was added to aqueous solutions of different pH
values: pH 1.7 0.2 (citrate buffer), 3.5 0.2 (citrate buffer), 7.0 0.2 (phosphate buffer), and 11.5 (5 % Na2CO3).
These solutions were distilled and continuously extracted for
60, 120 or 240 min as described before. The extracts obtained
were dried with anhydrous sodium sulphate and concentrated
to about 1 mL using a Vigreux column.
One-dimensional gas chromatographic (1D-GC)
analysis
Separation and quantification of targeted aroma compounds
were performed on a 60 m 9 0.32 mm i.d. fused-silica

capillary column, coated with 0.25 lm polyethylene glycol


as stationary phase (DB-Wax, J&W, Folsom, CA, USA).
Identification was via retention time and linear retention
index (benzaldehyde 1516; linalool, 1552; 1-octanol 1565).
Temperature was programmed from 40 C (5-min hold) with
4 min-1 to 230 C (25-min hold). Split injection was performed at 210 C, detection was by flame ionization (FID) at
240 C. Carrier gas used was hydrogen at 105 kPa constant
inlet pressure. A C.E. Instruments (today ThermoFisher
Scientific, Dreieich, Germany) 8575 Mega II series gas
chromatograph was used. Instrument control and data
acquisition were done with the Chromcard software (ThermoFisher). Quantification of linalool and benzaldehyde was
based on the internal standard 1-octanol, considering the
respective response factors.
Multidimensional gas chromatographic (MDGC)
analysis
A MDGC system that in literature is described as Moving
Capillary (Column) Stream Switching system (MCSS)
was used for heart-cut multi- (actually two-) dimensional
gas chromatography [37]. The system was originally
obtained from C.E. Instruments and is nowadays distributed by Brechbuhler AG (Schlieren, Switzerland). Details
on its operation and optimization have been described in an
earlier publication [38]. Basically, two C.E. Instruments
8000 series GC systems were coupled through a heated
transfer line, which was set to 200 C. The first-dimension
(1D; achiral pre-column) column consisted of a 38 m 9
0.25 mm i.d. fused capillary, coated in-house with a
polyethylene glycol stationary phase (Innowax, Portland,
OG, USA). Standard procedures for static coating were
followed [39], yielding a film thickness of 0.5 lm. Carrier
gas used was hydrogen at constant inlet pressure of
150 kPa (P1; for details see [38]), and 80 kPa (P2).
Detection was via FID, set to 240 C, and injection was in
splitless mode (45 s) at 210 C. A phenylmethyl-deactivated fused-silica capillary (0.75 m 9 0.25 mm i.d.) connected the GC oven of this 1D column with the separation
column in the second-dimension (2D) oven. This 2D (main)
column consisted of a phenylmethyl-deactivated 30 m 9
0.25 mm i.d. fused-silica capillary, coated in-house with a
mixture of 25 % heptakis (2,3-di-O-methyl-6-O-tertbutyldimethylsilyl)-b-cyclodextrin (synthesized according
to procedures described earlier [40, 41]) as the chiral stationary phase and 75 % SE 54 (RiedeldeHaen, Seelze,
Germany), yielding a film thickness of 0.25 lm. Temperature was programmed from 70 C (20-min hold) with
2 min-1 to 130 C and then with 10 min-1 to 210 C
(10-min hold). The actual inlet pressure (hydrogen) was
given at the read-out on the pressure gauge P3 and was at
78 kPa. With linalool having a retention time of 25.2 min

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(on the achiral 1D column), the cut interval was set to


25.1525.45 min.

Results and discussion


Linalool concentration and its ratio
versus benzaldehyde in cocoa beans
Concentrations of linalool and benzaldehyde as well as
their concentration ratios are summarized in Table 1. The
quantifications were based on the use of 1-octanol as
internal standard. This compound has been described as
constituent of roasted cocoa powder [42]; however, owing
to the low concentrations reported (00.044 mg kg-1;

mean: 0.019 mg kg-1), this occurrence was not expected


to hamper the quantification of the major target compounds.
Flavour-grade cocoas from Ecuador (Arriba) exhibited contents of linalool between 1.2- and 3.6 mg kg-1 peeled cocoa
beans. They were considerably higher than those found in
basic cocoas (0.20.8 mg kg-1). However, cocoa beans from
Venezuela and Amazonia, which had been traded as flavourgrade cocoas, showed linalool concentrations within the
range of the basic-grade cocoa materials. According to literature [9, 11], a linalool/benzaldehyde concentration ratio of
0.3 or higher is related to flavour-grade cocoas, whereas a
lower value represents basic-grade cocoas. For the sample set
available in this study, the low ratios of linalool/benzaldehyde (0.10.2) observed for the basic-grade materials and the
higher ratios (0.31.7) determined for the flavour-grade beans

Table 1 Analytical data for linalool and benzaldehyde in roasted and unroasted cocoa beans
Grade

Provenience

Roasting
(C)

Basic
Basic

Ghana
Ivory Coast

(min)

(S)-(?)-Linalool
(%)

Linalool
(mg kg-1)

Benzaldehyde
(mg kg-1)

Ratio lin./
benz.

Replicates
(n)

Raw

96.7 0.5

0.8 0.1

4.1 0.6

0.2 0.1

165

25

95.5 0.9

0.5 0.2

3.4 0.5

0.2 0.03

Raw

96.5 0.1

0.7 0.4

3.9 1.0

0.1 0.1

110

25

95.7 0.5

0.5 0.02

3.9 0.1

0.1 0.01

135

25

96.2 0.5

0.5 0.1

3.9 0.4

0.1 0.03

New
Guinea

Raw

96.3 0.6

0.2 0.05

2.5 0.5

0.1 0.0

110

25

97.1 0.2

0.3 0.1

2.7 0.2

0.1 0.02

135

25

97.1 0.2

0.3 0.1

3.2 0.1

0.1 0.1

Flavour

Amazonia

Raw

96.7 0.5

0.5 0.1

2.9 0.2

0.2 0.0

165

25

94.6 1.4

0.9 0.8

2.6 0.5

0.4 0.4

Flavour

Arriba (A)

Raw

94.0 1.4

3.6 0.5

2.2 0.4

1.7 0.4

165

25

92.1 0.6

3.0 1.2

1.9 0.2

1.5 0.6

Flavour

Arriba (A)a

140

30

92.0

n.d.b

n.d.

n.d.

Flavour

Arriba (B)

Raw
145

20

95.3 0.4
94.8 2.3

1.2 0.2
1.7 0.3

4.5 0.4
4.4 0.4

0.3 0.1
0.4 0.1

5
5

Flavour

Arriba (C)

Raw

95.3 0.4

2.9 0.9

3.8 0.4

0.7 0.2

5c (3)d

Basic

Flavour

Undefined

Undefined

Venezuela

Colombia

Jamaica

110

25

94.2 0.8

1.4 0.9

3.7 0.5

0.4 0.2

135

25

94.1 0.4

2.9 1.0

3.2 0.5

1.0 0.5

Raw

96.1 0.5

0.3 0.03

3.8 0.5

0.1 0.02

110

25

96.4 1.0

0.5 0.1

3.5 0.1

0.1 0.04

135

25

95.6 2.0

0.4 0.1

3.7 0.3

0.1 0.03

Raw

97.1 0.1

0.8 0.2

1.8 0.3

0.5 0.2

110

25

97.5 0.3

1.2 0.2

1.9 0.3

0.7 0.2

4
4

135

25

97.5 0.2

1.1 0.04

2.3 0.2

0.5 0.1

Raw

96.7

0.5

2.5

0.2

110

25

96.4 0.3

0.4 0.1

2.4 0.3

0.2 0.04

135

25

96.9 0.1

0.4 0.1

3.1 0.2

0.2 0.04

Sample preparation with liquidliquid extraction

Not determined

Replicates for enantio-MDGC

Replicates for quantitative determination of linalool and benzaldehyde

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from Arriba were in agreement with this correlation. However, the low ratios (0.2 and 0.1) obtained for the flavourgrade cocoas from Amazonia and Venezuela demonstrate the
potential limitations of this approach. An unequivocal
assignment of cocoa bean materials to one of the quality
grades solely on the basis of the ratio of the concentrations of
linalool and benzaldehyde seems questionable.
Influence of sample preparation on enantiomeric
composition
Considering the known instability of linalool in acidic
medium, several control experiments were performed.
First, (R)-linalool of known enantiomeric purity was
refluxed at different pH values; subsequent determination
of the enantiomeric composition by MDGC demonstrated
that linalool undergoes racemization only under acidic
conditions (Table 2). The basic mechanism of this process
is the previously described proton-catalysed 1-alken-3-ol/
2-alken-1-ol rearrangement [32, 43, 44]. In order to verify
the suitability of the simultaneous steam distillation
extraction (SDE) method, linalool of known enantiomeric
purity was analysed after SDE under extreme pH conditions (from pH 1.7 to 11.5) and different extraction periods
(Table 2). Comparable to the first experiments, linalool
only tends to racemize under acidic conditions. However,
when comparing a reflux period of 1 h with SDE for 1 h at
the same pH (3.5), considerably less racemization was
found with the SDE method. Even a prolonged period of
24 h for SDE at this pH did not cause a significantly higher
degree of racemization (data not shown). This may be
explained, as the proton-catalysed racemization occurs in
the aqueous phase, and most of the linalool is extracted
within the first 30 min [36].
Liquidliquid extraction of one of the cocoa samples
(Arriba) yielded identical results as SDE at pH 7, thus
proving the suitability of this isolation technique in conjunction with the determination of the enantiomeric distribution of linalool. The repeatability of the SDEMDGC
analysis of the enantiomeric distribution of linalool was
determined on a roasted (Amazonia) cocoa beans sample
(n = 5), resulting in 95.2 0.2 % (S)-(?)-linalool.
Enantiomeric distribution of linalool in cocoa beans
and cocoa products
The analysis of the linalool fraction from an aroma extract
of roasted cocoa beans (from Venezuela) via multidimensional GC is presented in Fig. 1. The evaluation of the
enantiomeric distribution in cocoa beans revealed a consistently high preponderance of (S)-linalool (Table 1). In
all cocoa samples analysed, linalool was found well above
90 % in favour of the (S)-enantiomer. Furthermore, the

Table 2 Influence of sample preparation conditions on optical purity


of (R)-(-)-linalool
pH

Time (min)

(R)-(-)-Linalool
(%)

(S)-(?)-Linalool
(%)

3.5

67.1

32.9

7.0
9.0

99.3
99.3

0.7
0.7

1.7

60

62.0

38.0

3.5

60

92.0

8.0

3.5

240

91.0

9.0

7.0

60

99.3

0.7

7.0

120

99.3

0.7

7.0

240

99.3

0.7

11.5

60

99.3

0.7

LEa

SDE

The enantiomeric purity of the reference (R)-(-)-linalool had prior


been determined by GC with 99.3 %
a

Liquid extraction after 1 h reflux


32

A*

(mV)
25.6
19.2
12.8

cut

6.4

24

26

48

(min)

42

(min)

0
0

12

24

Linalool

36

(S)
OH

OH

(R)
34

36

38

40

Fig. 1 Enantioselective MDGC analysis of linalool in a typical cocoa


bean (Venezuela) SDE extract: a achiral pre-column, a* cut-window,
and b chiral (main) column. Conditions as described in Materials
and methods section

data obtained for various samples processed under different


temperature and time conditions revealed no significant
influence on the original enantiomeric distribution of
linalool.
Cocoa beans of different origin showed minor differences regarding their enantiomeric distribution. However,
the range is very narrow; for example, from 94 % (S)-linalool in the Arriba samples to about 97 % in cocoa beans

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Table 3 Contents and enantiomeric distributions of linalool in
commercial cocoa powders and chocolates
Linalool
(mg kg-1)

R (%)

\0.1

13

87

Brand B (contains West African cocoa)


0.2
Brand C
\0.1

7
9

93
91

Product

S (%)

Cocoa powders
Brand A

Brand D

\0.1

92

Brand E

\0.1

10

90

Brand F

0.3

11

89

Brand G

0.2

10

90

Brand H (from African cocoa)

0.2

95

Brand A, with cocoa chunks

0.1

93

Brand B, Zartbittera

0.3

94

Brand C, Halbbittera

0.2

94

Brand D, Mocha

\0.1

11

89

Brand E, Mochab

\0.1

11

89

Brand F, whole milka

\0.1

91

Chocolates

Declaration (German) representing percentage of cocoa; see further


information: http://www.theobroma-cacao.de

Declaration: With Flavour Grade Cocoa

from Colombia. With respect to the enantiomeric composition of linalool, no difference was found between flavourand basic-grade cocoa beans.
The data for the enantiomeric distribution of roasted and
unroasted cocoa beans are almost identical; they demonstrate that there is no significant influence on the original
enantiomeric distribution of linalool during the roasting
process, even though the milling of the beans to a size of
2 mm before the thermal treatment can be considered as
worst-case scenario compared to industrial conditions.
The results obtained for commercial cocoa powders and
chocolates (Table 3) indicate that even technological procedures used in the production of cocoa products such as
alkalisation (cocoa powders) or conching (chocolate) do
not change the original distribution significantly. Except
for a small increase of the proportion of the (R)-enantiomer, observed in particular for the Brand A sample, the
high excess of (S)-linalool can still be found in such
products.
Interestingly, the absolute concentration of linalool does
not change significantly from raw to roasted cocoa beans.
This has also been described recently in a comprehensive
study following changes of key aroma compounds of
Criollo cocoa beans during roasting [8]. Linalool is known
to be often found not only in its free form, but also bound
to glycosidic precursors, as has been described for Vitis
vinifera varieties [45], but also for others. If this would be

123

the case for cocoa beans, liberation (either enzymatic or


acidic hydrolysis) should be expected under certain sample
preparation or technological processing steps. As an
example, an increase of linalool has been observed during
SDE of cherry (Prunus) juices and during jam production,
which was caused by hydrolysis of the corresponding
glycosides during the heat treatment [46]. On the other
hand, roasting conditions, thus applying a relatively high
temperature over a certain period of time, should decrease
the linalool concentration, simply due to its volatility. In
our situation, no increase in linalool concentration was
found, for example, for SDE at pH 3.0 and for a period of
60 or 120 min (data not shown). Even under conditions,
such as a roasting temperature of 165 C (rather high for
flavour-grade cocoas), as applied to Arriba or Ghana
samples, no significant change was found (Table 2). On the
other hand, a loss in linalool concentration has been
described in studies comparing dark chocolate samples
before and after conching [47].

Conclusion
Linalool could be successfully enantiodifferentiated in
SDE extracts from cocoa beans or products thereof by
MDGC and a chiral stationary phase. SDE conditions with
a neutral-buffered solution, allowed enantiodifferentiation
without the risk of partial racemization, which may occur
under acidic conditions. It was found that cocoa beans of
both flavour- and basic-grade varieties, as well as from
different origins, have a high proportion of (S)-linalool of
usually well above 90 %. This still remains in commercial
products, indicating that technological processes such as
roasting or alkalisation have no detrimental effect on the
high enantiomeric excess of linalool. Interestingly, the
linalool content does not change significantly from raw to
roasted beans. If this can be explained by a balance
between generation from potential precursors and losses
due to the roasting process still remains an open question.
Considering the differences in odour properties and odour
thresholds reported for the enantiomers of linalool, future
studies should also focus on the sensory contribution of the
(S)-enantiomer of this monoterpene alcohol to the cocoa
flavour.
Acknowledgments The authors gratefully acknowledge Claudia
Preis, Carsten Stein, and Lilian Simoes dos Santos for their contribution and technical assistance in this work. We also thank the
Fraunhofer Institut fur Lebensmitteltechnologie und Verpackung
(Freising, Germany), the Bundesverband der Deutschen Suwarenindustrie e.V. (Koln, Germany), HACHEZ (Bremen, Germany),
Kraft Jacobs Suchard R&D, Inc. (Munchen, Germany), as well as the
Verein der am Rohkakaohandel beteiligten Firmen e.V. (Hamburg,
Germany) for donating defined cocoa samples.

Eur Food Res Technol

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