Eur Food Res Technol (2006) 223: 261266

DOI 10.1007/s00217-005-0193-9
ORIGINAL PAPER
Dirk W. Lachenmeier Stephan G. Walch
Waltraud Kessler
Using experimental design to optimise precision of steam distillation
for determining alcoholic strength in spirits
Received: 23 August 2005 / Revised: 24 October 2005 / Accepted: 26 October 2005 / Published online: 23 December 2005
C
Springer-Verlag 2005
Abstract An experimental design, in combination with
oscillation-type densimetry is introduced as a novel pro-
cedure to optimise steam distillation for the determination
of alcoholic strength in different types of alcoholic bev-
erages. A central composite design was chosen to study
the effects of variation in levels of receiver volume (25
and 50 ml), steam power (30100%), distillation time (20
140 s) and sample volume (550 ml). Three commercial
spirit drinks with different alcoholic strengths (4.9, 35.3,
and 54.7% vol.) were compared on two automated steam
distillation devices of different manufacturers by separately
completing the experimental design for each. The mod-
els tted for the prediction of the alcoholic strength, as
indicated by an r
2
value of more than 0.93. Various sur-
face plots were generated to describe the relationship be-
tween operating variables and predicted alcoholic strength.
Optimum conditions were 50 ml receiver volume, 70%
steam power, 100 s distillation time and 25 ml sample vol-
ume. Slight deviations from the optimum did not result in
substantial decrease of alcoholic strength. Validation ex-
periments carried out under predicted conditions showed
excellent correspondence to the reference procedure
(R=0.999).
D. W. Lachenmeier ()
Chemisches und Veterin aruntersuchungsamt (CVUA)
Karlsruhe,
Weienburger Str. 3,
D-78187 Karlsruhe, Germany
e-mail: Lachenmeier@web.de
Tel.: +0721-926-5434
Fax: +0721-926-5539
S. G. Walch
Zentrales Institut des Sanit atsdienstes der Bundeswehr Koblenz,
Andernacher Str. 100,
D-56070 Koblenz, Germany
W. Kessler
Hochschule Reutlingen, Fakult at Angewandte Chemie,
Alteburgstr. 150,
D-72762 Reutlingen, Germany
Keywords Ethanol
.
Alcoholic strength
.
Steam
distillation
.
Oscillation-type densimetry
.
Experimental
design
.
Optimisation
.
Response surface methodology
Introduction
Within the work of ofcial food control, alcoholic strength
by volume is one of the most important parameters in spirit
drink analysis. In comparison to other countries, the Euro-
pean Union allows only a marginal tolerance of 0.3%vol.
concerning the indication of the actual alcoholic strength
by volume in the labelling [1]. In 2004, more than 15%
of all samples of liqueurs and fruit spirits analysed in the
German federal state of Baden-W urttemberg had to be ex-
cluded due to faulty labelling of the alcoholic strength [2].
Exceeding the tolerated limits can have grave consequences
for the manufacturers such as nes, high costs of recall and
relabelling of a production lot. If, on the other hand, the
alcoholic strength of the products is adjusted too high, an
economic loss will be the consequence.
Until recently, distillation with subsequent pycnomet-
ric determination of the density has been the reference
method for the determination of alcoholic strength in spirit
drinks. Pycnometers were generally considered to provide
the greatest accuracy, but the method is time-consuming
and requires special training of personnel if reproducible
results are to be obtained because of a relatively greater op-
portunity to cause experimental errors in making the weight
measurements necessary for pycnometry [3]. Automated
methods such as gas chromatography or liquid chromatog-
raphy could not replace pycnometry as a reference method
because they are instrumentally complex and therefore
more expensive. Furthermore, handling required trained
personnel, and some methods did not have the required
accuracy [48]. Secondary analytical technique that use
near-infrared spectroscopy or Fourier transform infrared
spectroscopy in combination with chemometric techniques
may be used for a fast screening of alcoholic strength but
also do not qualify as reference procedures [6, 7, 911].
262
In the Eighties, electronic densimetry, which is based
on electromagnetically-induced oscillation of a U-shaped
glass tube, was introduced into the analysis of alcoholic
strength, showing similar performance in terms of accuracy
and precision in comparison to established methods like
pycnometry, hydrostatic balance or hydrometry [3, 1214].
This is nowadays the state-of-the-art method, because it is
time-saving and cost-saving as well as simple to perform
[3, 15]. In consequence, oscillation-type densimetry has
been introduced into the European community as reference
method for the analysis of spirit drinks [16].
The remaining limiting factor is the conventional dis-
tillation step commanded in the reference methods. This
distillation is time-consuming and requires high personnel
expenditures. Steam distillation was recently introduced as
a promising alternative for determining alcoholic strength
in all kinds of spirit drinks [17, 18]. With the carrier vapour,
the distillable part of a mixture is separated from the non-
volatile residue. Simultaneously a boiling point depression
occurs and a thermally mild treatment is achieved. By pass-
ing steam into the spirit sample, the alcohol is expelled
and signicantly shorter times of distillation are achieved
in comparison to conventional distillation. In combination
with oscillation-type densimetry, the procedure takes less
than 8 min per sample.
In this study, the inuence of all basic operation parame-
ters of the steam distillation device such as steam power of
the steam generator, time of distillation, as well as sample
volume and receiver volume on the outcome of the alco-
holic strength was examined. As these settings inuence
each other, show interactions and, additionally, a partly
nonlinear behaviour, the possibility to give one setting for
all feasible alcoholic strengths and the transferability be-
tween steam distillation devices of different manufacturers
was investigated.
Materials and methods
Instrumentation
The automated steam distillation was accomplished with
the Gerhardt Vapodest 30 (C. Gerhardt, Fabrik und Lager
chemischer Apparate, Bonn) as well as the B uchi K-355
(B uchi, Flawil, Switzerland). The devices were coupled to
tanks lled with distilled water. Before every start up the
steam generators were preheated with a water sample at
full steam power (according to the manufacturers instruc-
tion). For the tempering of the sample the heating circu-
lator bath DC10-W26 (Haake, Karlsruhe, Germany) was
used. The determination of density was accomplished with
the density meter DE51 with sample pump ASU-DE by
Mettler-Toledo (Giessen, Germany). The instrument was
adjusted with air and water according to the manufacturer.
The adjustment was checked daily using certied water
standards (=0.99820 at 20

C). The sample temperature
in all measurements was adjusted to 20

C. The alcoholic
strength was calculated automatically from the measured
density using the stored ofcial alcohol table data. Between
measurements, the connecting tubes were purged with air
and cleaned with distilled water. Before shutdown of the
system, the hoses were purged with acetone and air until
they were dry.
The reference method for determination of the alcoholic
strength in spirit drinks was applied without modica-
tion as prescribed in the Commission Regulation (EC) No.
2870/2000 laying down Community reference methods for
the analysis of spirit drinks [16]. The pycnometers and
the distillation apparatus were purchased at Paris, Technis-
che Glasbl aserei (Karlsruhe, Germany) after the reference
methods specications.
Sample preparation and measurement
The sample was temperated in a water bath at 20

C. Fol-
lowing this, the sample was pipetted into a 250 ml Kjel-
datherm digestion tube. Rest of the sample sticking to the
edge of the tube were rinsed down with distilled water. Sub-
sequently the tube was clamped in the distillation device.
After placing a graduated ask lled with 3 ml of distilled
water under the distillate outlet tubing, the programme was
started, and the distillation was automatically performed.
After termination, the receiver and the tube were replaced
and the hoses were rinsed with distilled water to be ready
for the next sample. The graduated ask with the distillate
was temperated and lled up to the calibration mark. The
alcoholic strength was determined with the oscillation-type
density meter.
Optimisation using experimental design
The following operation parameters of steam distillation
were optimised with regard to maximumalcoholic strength:
receiver volume (25 and 50 ml), steam power (30100%),
distillation time (20140 s) and sample volume (550 ml).
To cover the usual operation range, measurements with
samples of different alcoholic strengths were carried out.
A herbal liqueur with reference alcoholic strength of
35.28% vol., an alcopop drink with a reference value of
4.88% vol. and an absinthe with a high alcoholic strength
of 54.69% vol. were tested. All test series were performed
on both the Gerhardt and the B uchi device.
In order to nd the optimal working settings with min-
imum amount of experiments, the information contained
in each experiment and furthermore the relation between
the experiments has to be fully exploited by discovering
interactions and nonlinear dependence. The important step
was to set up an orthogonal experimental design space.
This was done with a central composite design [19, 20].
The chosen levels of the variables are shown in Table 1.
Studying three factors at ve levels in a complete design
would require 5
3
or 125 samples, whereas using a central
composite design requires only 20 samples and still totally
covers the experimental space and allows to calculate all
interactions and nonlinearities. The calculations were done
using the Software Package Design Expert V6 (Stat-Ease
Inc., Minneapolis, USA).
263
Table 1 Levels of variables used in the central composite design
Variable Name Levels
A Steam power 30, 44, 65, 86, 100
B Distillation time 20, 44, 80, 116, 140
C Sample volume 5.0, 14.1, 27.5, 40.9, 50
To further check the applicability and trueness, 54 sam-
ples from the study stack of the CVUA Karlsruhe (alco-
holic strength range 2.055.5% vol.) were measured with
both densimetry following steam distillation and the py-
cnometric reference method under within-laboratory re-
producibility conditions (different operators on different
days).
Results and discussion
Optimisation of receiver volume
Graduated asks with volumes of 25 and 50 ml were avail-
able as receivers for the distillate. As a change in outlet
volume obviously leads to a complete deviation of the other
parameters, the rst experiments were aimed to nd the op-
timal receiver volume before the optimisation of the other
parameters. As can be seen in Fig. 1, the experiments with
the 25 ml receiver led to higher deviations especially at low
distillation times, which can be attributed to higher pipet-
ting and dilution errors. In contrast, the 50 ml receiver had
a higher operating range with reproducible results above
80 s of distillation time. This conrms previous empiri-
cally determined results that a 50 ml measuring ask is best
[17, 18]. All further experiments were, therefore, conducted
using 50 ml receivers.
Fig. 1 Comparison between 25 and 50 ml graduated asks as re-
ceiver for the alcoholic distillate (B uchi instrument). To reach com-
parability, the experiments for the 25 ml receiver were conducted
with half the sample volume and half the distillation time com-
pared to those used for the 50 ml receiver. At longer distillation
times than 70 or 140 s, the receiver is lled above the calibration
mark
Optimisation of steam power, sample volume
and distillation time
The experiments of the central composite design were eval-
uated using Analysis of Variance (ANOVA) to nd the sig-
nicance of the linear, quadratic and interaction terms in
the response surface models. A very important step was to
check the signicant models for consistency by looking at
the lack of t and possible outliers. In all six designs the
extreme settings with a very low distillation time and those
with a very high sample volume showed to be clear outliers
and had to be left out of the analysis.
For both instrumental setups (Gerhardt and B uchi) it is
possible to nd the optimal working range. By means of
response surface analysis the regression coefcients of the
model are determined and the statistical ANOVA approach
calculates the individual signicance of each coefcient.
Table 2 lists the regression coefcients. The factors are
given in coded values, which make the six models directly
comparable between each other and offer the opportunity
to nd the importance of each regression termin the model.
Signicant differences between the two steam distillation
devices were found. The B uchi instrument is independent
of the variation in steam power (A), whereas at the Ger-
hardt instrument, the variation in steam power only slightly
inuences the distillation of alcopop, highly inuences that
of absinthe and has no inuence on herbal liqueur. This dif-
ference can easily be explained by the fact that the B uchi
instrument has a signicantly higher steampower (2200 W)
than the Gerhardt instrument (1600 W).
The changes in distillation time (B) and sample volume
(C) are very decisive in all the cases for measuring the
alcoholic strength. The distillation time has a quadratic in-
uence (B
2
) for all instruments. The Gerhardt instrument
is again different to the B uchi instrument as steam power
shows an additional quadratic inuence (A
2
). The interac-
tion between distillation time and sample volume (BC) has
a lower inuence on the Gerhardt instrument than for the
B uchi one.
With these models, it is possible to dene an optimum
working area for each instrument for determining the al-
coholic strength of unknown spirits. In Figs. 2 and 3, the
overlay response surface plots for the optimal working area
for the two different steam distillation devices is shown for
three samples with very different alcoholic strengths, re-
spectively. For easy visualization, the region tting the py-
cnometric reference value of alcoholic strength is marked
white in the response contour.
The response surface plots showclearly that the optimum
working area is not much inuenced by the steam power,
that a sample volume between 15 and 30 ml and a distilla-
tion time between 90 and 110 s give best results for both
instruments and spirits of different alcoholic strengths. The
results are summarized in Table 3.
Altogether, the method proved to be very rugged. The
optimal settings for either instrument are independent of
the varying inuences of the parameters including the al-
coholic strength of the sample and the type of steam distil-
lation device in a wide range. The steam distillation device
264
Table 2 Regression
coefcients in coded values for
the optimisation of steam
distillation with different
alcoholic beverages on different
instruments
Alcopop 4.9% vol. Herbal Liqueur 35.3% vol. Absinthe 54.7% vol.
Regression coefcient Gerhardt B uchi Gerhardt B uchi Gerhardt B uchi
Intercept 5.22 4.89 34.53 35.06 54.04 54.58
A 0.15

no effect no effect no effect 4.29

no effect
B 0.34

0.27

2.66

1.57

4.12

3.75

C 0.21

0.21

1.23

1.50

3.26

4.20

A
2
0.09

no effect 1.20

no effect 2.59

no effect
B
2
0.17

0.12

1.20

0.93

1.85

1.65

C
2
no effect 0.22

1.53

0.83

no effect 1.93

AB 0.13

no effect 0.33

no effect 3.01

no effect
AC no effect no effect 0.45

no effect 1.21

no effect
BC 0.13

0.15

0.42

1.01

1.14

3.17

r
2
0.918 0.930 0.998 0.994 0.991 0.997
Standard deviation 0.12 0.13 0.17 0.17 0.67 0.41
A steam power, B distillation
time, C sample volume

P0.05;

P0.01;

P0.001
A: steam power [%]
B
:

d
i
s
t
i
l
l
a
t
i
o
n

t
i
m
e

[
s
]

40 55 70 85 100
50
65
80
95
110
Herbal liqueur: 35.28
Alcopop: 4.88
Absinthe: 54.69
C: sample volume = 25 ml
B: distillation time [s]
C
:

s
a
m
p
l
e

v
o
l
u
m
e

[
m
l
]

50 65 80 95 110
10
20
30
40
50
Herbal liqueur: 35.28
Alcopop: 4.88
Absinthe: 54.69
A: steam power = 76%
Fig. 2 Response surface plots
showing the optimal working
area (white) of steam power,
distillation time and sample
volume for different alcoholic
strengths determined using the
B uchi instrument (lines show
reference values for herbal
liquor, alcopop and absinthe
sample)
A: steam power [%]
B
:

d
i
s
t
i
l
l
a
t
i
o
n

t
i
m
e

[
s
]
40 55 70 85 100
50
65
80
95
110
Herbal liqueur: 35.28
Absinthe: 54.69
C: sample volume = 25 ml
Alcopop: 4.88
B: distillation time [s]
C
:

s
a
m
p
l
e

v
o
l
u
m
e

[
m
l
]
50 65 80 95 110
10
20
30
40
50
Herbal liqueur: 35.28
Alcopop: 4.88
Absinthe: 54.69
A: steam power = 76%
Fig. 3 Response surface plots
showing the optimal working
area (white) for steam power,
distillation time and sample
volume for different alcoholic
strengths determined using the
Gerhardt instrument (lines show
reference values for herbal
liquor, alcopop and absinthe
samples)
Table 3 Results of method
optimisation for Gerhardt and
B uchi instruments
Admissible range Optimised setting
Parameter Gerhardt B uchi Overall
Reveicer volume (ml) 50 50 50
Steam power (%) 5580 40100 70
Distillation time (s) 90115 70110 100
Sample volume (ml) 1530 1535 25
265
Table 4 Linear correlation between analysis results using the refer-
ence method (pycnometry with standard distillation) and oscillation-
type densimetry with steam distillation for the determination of alco-
holic strength in spirit drinks under within-laboratory reproducibility
conditions (B uchi instrument)
Parameter Result (n=54)
Slope (LCI, UCI) 0.998 (0.992, 1.004)
Intercept (LCI, UCI) 0.011 (0.189, 0.210)
Correlation coefcient 0.99979
Probability <0.0001
Mean bias (%vol.) 0.02
is programmed with the prescribed parameters, which are
monitored during the operation. If deviations occur in the
process (e.g. shortfall of the steam power) warnings are
displayed. Due to its higher steam power, the B uchi instru-
ment has a slightly larger optimal range, but the Gerhardt
instrument also gives accurate results as previous validation
studies have shown [17, 18].
Comparison to the reference procedure
Pycnometry is generally regarded as the reference proce-
dure for the determination of alcoholic strength in spirits.
Despite the disadvantages described in the introduction, the
pycnometric values can be viewed as empirical reference
values to which other methods have to agree.
To prove the applicability of the new method, several
spirits were analysed using steam distillation with the
determined optimal working parameters as well as with
the reference procedure. The linear correlation between
both methods was excellent, with a correlation coefcient
higher than 0.999 (Table 4). Since slope and intercept
encompass the theoretical values, no constant or propor-
tional differences between the two procedures could be
proven except for random errors. The deviation of the
results obtained by oscillation-type densimetry in com-
parison to pycnometry was on average only 0.02% vol..
The excellent linearity in the examined range between 2
and 55.5% vol. makes the method equally suitable for the
analysis of mixed spirit drinks with only minor alcoholic
strength as well as for high-proof spirit drinks, such as
absinthe.
Advantages of steam distillation in combination
with oscillation-type densimetry
In accordance to the reference method, the distillation re-
mains the basis even for this new, time-saving analysis.
During method optimisation, it proved to be advantageous
to distil about 25 ml of sample into a 50 ml graduated
ask as a receiver. In contrast to the reference method, the
distillate is diluted during this process. However, the com-
parison with pycnometry showed that the calculation error
is negligible. The use of steamfor the distillation of alcohol
is fast, cost-saving and requires only a minimum of spe-
cial knowledge of the laboratory personnel. In contrast to
pycnometry, untrained personnel could easily achieve ac-
curate results. Therefore, it could be the method of choice
for governmental laboratories as well as for spirit manu-
facturers and distilleries. A further advantage is the fact
that it can be used in the analysis of uids that possess a
high viscosity and boiling point and which usually tend to
scorching and spattering in conventional distillation (e.g.
emulsion liqueurs). Through the simultaneous heating and
mixing effect of the steam injection, a good intermixture is
assured, while at the same time a scorching of the sample is
nearly impossible and was never observed. Of course, the
cleaning of the used glass devices is considerably easier
with this method.
Conclusions
The shortened time of steam distillation (approx. 2 min) is
a signicant advantage compared to the classic distillation
(approx. 30 min). Less than 8 min per sample are needed
for the complete analysis including steam distillation and
densimetry, which means that a high sample throughput of
nine samples per hour is possible. Furthermore, a complete
automation of both steams distillation and densimetry is
possible using autosamplers, so that any operator dependent
inuence can be totally eliminated and the efciency is
further enhanced. Both the possibility of high-throughput
and the marginal cleaning effort lead to a cost reduction in
personnel and therefore further prove that this method is
superior to others.
Acknowledgements The skilful technical assistance of S. Gonzalez
and H. Heger is gratefully acknowledged. The authors thank J. M uller
of B uchi Labortechnik (Flawil, Switzerland) for providing the steam
distillation device. Presented in part at EURO FOOD CHEM XIII
(Hamburg, Germany).
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