Eur Food Res Technol (2003) 217:7073
DOI 10.1007/s00217-003-0700-9
ORIGINAL PAPER
Ulrich Krings Dattatreya S. Banavara Ralf G. Berger
Thin layer high vacuum distillation to isolate the flavor
of high-fat food
Received: 9 December 2002 / Revised: 12 February 2003 / Published online: 29 March 2003

Springer-Verlag 2003
Abstract A modified vacuum distillation apparatus to
isolate volatiles from lipid food matrices was systemat-
ically evaluated using neutral synthetic oil (Miglyol 812)
spiked with 14 flavor compounds. Thin layer high
vacuum distillation showed good recovery for most of
the flavor constituents of the model mixture. The recovery
was linearly correlated with the product of the octanol/
water partition coefficient, logP and boiling point when
the lactones were excluded. The same linear relationship
was found for lactones, but the slope of the regression line
was much steeper. The mean recovery of d-dodecalactone
was 5.1%, indicating the limit of the method.
Keywords Vacuum distillation High-fat food Flavor
recovery Octanol/water partition coefficient Lactones
Introduction
The primary principles used in isolating aroma con-
stituents are volatility, partition, or both in combination
[1]. With increasing lipid content of a food, the release of
volatiles into the gaseous phase is considerably decreased
[2]; hence, even dynamic headspace sampling of flavor
constituents often does not reach the limit of detection of
less volatile flavor compounds. Increased temperature
increases partial pressures, but holds the risks of non-
representative flavor composition and artifact formation.
Exhaustive extraction using non-polar solvents yields
solutions containing all more or less hydrophobic volatile
and non-volatile constituents of the food. The partition
coefficients of flavor compounds between lipids and
extraction solvents are close to one. Thus, lipoids have to
be separated from the flavor constituents prior to further
analyses. Variants of simultaneous distillation and ex-
traction (SDE) have answered the purpose for some
U. Krings D. S. Banavara R. G. Berger ())
Institut fr Lebensmittelchemie im Zentrum Angewandte Chemie,
Universitt Hannover,
Wunstorfer Strae 14, 30453 Hanover, Germany
e-mail: rg.berger@lci.uni-hannover.d
decades [3, 4]. Among the disadvantages of SDE are the
liberation of volatiles from antifoaming agents, greases
used in the joints and distilled water [5], and from the
oxidation of lipids, ester hydrolysis, Maillard reaction,
and sugar degradation [6, 7].
Weurman et al. [8] presented a vacuum distillation
method to transfer the flavor molecules at lower temper-
atures. A three-stage condenser was used [9] to condense
highly volatile flavor compounds under continuous vac-
uum pumping. The vacuum used in SDE by most workers
was not below 5,000 Pa, as compiled recently by
Chaintreau [10]. The original high vacuum transfer
(HTV) technique [8] was further refined by various
researchers [11, 12, 13, 14]. More recently, Engel et al.
[4] introduced a sophisticated apparatus for solvent
assisted flavor evaporation (SAFE) to improve the
recovery of flavor. The recoveries from the food matrices
or extracts with a high amount of fat (up to 50%) were
<40%, even when a high vacuum of 510-3 Pa was
applied. This lack of an easy and efficient isolation
procedure results in comparatively little knowledge of the
flavors of some lipid-rich foods.
Materials and methods
Distillation apparatus
The high vacuum distillation apparatus was based on the HTV
equipment used by earlier authors [11, 12, 13, 14]. Figure 1 shows
the entire equipment for performing the flavor evaporation and
trapping. Any liquid can be introduced as small droplets into the
high vacuum zone. Flavor samples were stored in a dropping funnel
connected with a jacketed coil condenser equipped with a vacuum
connection and connected to a 250 ml round bottom flask with
magnetic stirring. The round bottom flask was placed in a
thermostatted water bath and the coil of the condenser was
thermostatted with circulating water (Julabo F10, Julabo, Seelbach,
Germany). The apparatus was joined to a high vacuum system
(high vacuum pump E2M-07, turbo molecular pump EXC 120 and
active gauge controller, all BOC Edwards, Kirchheim, Germany)
across four serial cryogenic traps. Traps were placed into Dewar
vessels and either cooled by dry ice (first one) or by liquid nitrogen.
 71
of the coil the liquid, lipid moved downwards as a thin film with an
average flow velocity of about 0.5 ml min-1. The lipid film was
collected under continuous stirring in the round bottom flask. The
temperature of the water bath was finally increased to 50 C for
30 min to conclude the distillation process.

Analysis of recovered volatiles

Flavor constituents were rinsed out of the traps using 310 ml

pentane/diethyl ether (1:1.12) for each trap. The collected flavor
solutions were dried (Na2SO4) and concentrated to approximately
10 ml at atmospheric pressure and 40 C bath temperature using a
Vigreux column. g-Nonalactone (4.8 mg) was added to normalize
the injection volume.

GC-flame ionization detector analysis. The concentrated distillates

(1 l samples) were injected on-column. The instrument used was a
Fisons GC-8060 (Fisons, Mainz-Kastel, Germany) equipped with a
cool on-column injection port and a flame ionization detector
(FID)(250 C). Analytical conditions were as follows:
30 m0.32 mm i.d.0.25 m DB-WAX (J&W Scientific), hydrogen
as carrier gas at 48 cm s-1, and oven temperature: 45 C held for
5 min, to 150 C at 5 C min-1, to 240 C at 10 C min-1, held for
10 min.

GC-MS analysis. The purity and stability of model mixture

Fig. 1 Apparatus used for thin layer high vacuum distillation
(TLHVD) of lipids containing volatile flavors
constituents were analyzed by means of GC-MS. Of each sample,
1 ml was injected on-column. Instruments used were a Fisons GC-
8060 (Fisons, Mainz-Kastel, Germany) equipped with a cool on-
column injection port combined with a mass selective detector, MD
800 (Fisons). Analytical conditions were as follows: 30 m0.32 mm
i.d.0.25 m DB-WAX (J&W Scientific), helium as carrier gas at
38 cm s-1, same temperature program as used for GC-FID analysis,
detection in the electron impact mode with ionization voltage 70 eV,
continuous scanning from m/z 33 to 300, scan time 0.95 s, interscan
delay 0.05 s.

Thin layer high vacuum distillation procedure
Prior to distillation, each joint of the apparatus was sealed by a fine
film of high vacuum grease. The temperatures of the circulating
water and of the water bath were adjusted to 45 C. After a stable
high vacuum below 510-3 Pa was achieved, the flavor-containing
lipid sample was slowly introduced into the vacuum. A following
droplet was not allowed to enter the system until the system
reached a vacuum below 510-3 Pa again. Due to the tremendous
pressure gradient the droplets burst, and the non-volatile lipid was
sprayed onto the top of the inner coil of the condenser as a thin
film. Depending on the viscosity of the sample and the temperature
Model mixture. The 14 flavor compounds (dissolved in pentane/
diethyl ether, 1:1.12, v/v) varying in polarity (octanol/water
partition coefficient, logP) and boiling point in were dissolved in
50 ml of neutral synthetic oil (Miglyol 812, Sasol, Witten,
Germany) to a final concentration of 3.2 mg l-1 for each compound.
The same amount was added to 10 ml of pentane/diethyl ether and
used as an external standard (100% recovery). A compilation of
physicochemical properties is given in Table 1. To facilitate the
introduction of the sample into the thin layer high vacuum
distillation (TLHDV), 10% and 50% of solvent (pentane/diethyl
ether, 1:1.12, v/v), respectively, were added. Five replicates were
carried out and the recovery was expressed as the relative mean

Table 1 Physicochemical Flavor Molecular weight LogP Boiling point Supplier

properties of the model flavor g mol-1 C
constituents and standards [15,
16, 17]. A Fluka, Germany, Cyclopentanone 84.12 0.63 131 A
B Dragoco Gerberding, Pentan-1-ol 88.15 1.51 137 A
GermanyC ICN Biomedicals, 2(E)-hexenol 100.16 1.61 135 B
Germany, D Sigma-Aldrich, Heptanal 114.19 2.29 153 A
Germany Isobutyl acetate 116.16 1.77 116 B
2-Phenyl ethanol 122.17 1.56 215 C
2-Isobutyl thiazole 141.23 2.94 180 B
g-Octalactone 142.20 1.59 234 D
a-Terpineol 154.25 3.33 215 A
g-Decalactone 170.25 2.57 281a A
b-Damascenone 190.30 4.03 234 B
d-Dodecalactone 198.30 3.33 310a B
Valencene 204.35 6.3 245a A
b-Caryophyllene 204.34 6.3 256a B
a
Extrapolated
72
percentage (standard deviation) compared to the external standard
solution.

Results and discussion

The spiked Miglyol solution was subjected to TLHVD
(Fig. 1). The operating temperature was found to be best
at 45 C (data not shown). The recoveries of the
constituents of the model mixture are given in Table 2.
No significant difference between 10% (triplicate) and
50% (duplicate) solvent addition to the neutral oil was
observed (data not shown) with the exception of an
extensively enlarged distillation time. Hence, the results
were treated as one data set. The standard deviations of
five replicates were found to be less than 7.5%,
indicating a sufficient reproducibility of the whole
process. Advantages and disadvantages of the most
versatile extraction techniquesimultaneous extraction
of steam distillates by solventswere recently reviewed Fig. 2 TLHVD of model flavor constituents (see Table 2) dis-
by Chaintreau [10]. Recoveries of high boiling point solved in Miglyol; recovery plotted vs. boiling point and logP
compounds were found to be insufficient and application
of SDE to more lipoid matrices failed. Engel et al. [4]
improved the recoveries for less volatile compounds and tion with the recoveries obtained using TLHVD (Fig. 2).
filled the gap to more lipoid matrices using a solvent The recoveries tended to follow the boiling points more
assisted distillation (SAFE) under reduced pressure than the logP values. To take both parameters into
(510-3 Pa). Recoveries reported for a model flavor account, the product of boiling point and logP was
distributed in an oily matrix (50% fat) remained below calculated and correlated with the recovery of each
40% for all constituents. Some improvements achieved by compound (Table 2). Again, only a weak correlation was
TLHVD compared to SAFE are shown in Table 2. Good achieved for the whole data set. The lactones were the
recovery was not obtained for all compounds, particularly compounds that contributed most to the poor linear
the lactones. regression. Therefore, lactones and the other flavor
For accurate quantification, the reliable and well constituents were considered separately (Fig. 3). For the
established stable isotopic dilution method will remain separately treated groups of compounds, the recoveries
indispensable. However, this method is applicable to correlated well with the product of boiling point and logP.
compounds only which have been already identified in a The negative slope of the correlation curve was much
sample, and the identification, especially of trace con- steeper for the lactones indicating poor recoveries.
stituents, is in turn facilitated by enhanced high recoveries Compared to compounds of similar molecular weight
as provided by TLHVD. and functionality, lactones differ also in other parameters
The tendency of a flavor molecule to evaporate is such as boiling point and retention (Table 3). When
mainly expressed by its boiling point, whereas partition compared to the lactones the ethyl esters as well as the
behavior is usually expressed by its logP value. Neither acetates of the corresponding non-cyclic compounds with
logP nor the boiling point alone offered a close correla- the same carbon number exhibit much lower boiling

Table 2 Recovery of model Flavor b.p.LogP Recovery [%] Standard deviation Recovery %
flavor constituents dissolved in Mean of five replicates (n=5) SAFE [4]
Miglyol using TLHVD
Cyclopentanone 90.4 86.9 2.1
n-Pentanol 206.9 85.9 2.6
2(E)-Hexenol 217.4 86.1 4.7
Heptanal 350.4 85.2 1.2
Isobutyl acetate 205.3 85.6 1.8
2-Phenyl ethanol 335.4 75.2 5.6 12.8
2-Isobutyl thiazole 529.2 82.5 3.6
g-Octalactone 372.1 57.4 6.6
a-Terpineol 716.0 75.5 4.1
g-Decalactone 722.2 28.2 7.5
b-Damascenone 1,108 58.7 4.8 4.2
d -Dodecalactone 1,032 5.1 2.6
Valencene 1,543 54.9 0.6
b-Caryophyllene 1,613 57.4 5.6
 73

points and weaker retention on two GC-columns of

different polarities. However, a cyclic structure does not
explain this behavior, as cyclic and acyclic alcohols do
not differ much in these properties. In conclusion,
TLHVD enables sufficient recoveries for different classes
of volatile compounds from lipid-rich foods under mild
conditions, but remains problematic for non-polar high
boiling point compounds, especially for lactones.

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