LWT - Food Science and Technology 154 (2022) 112593

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LWT
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A simple and green method for determination of ethanol in liquors by the

conductivity measurement of the uncatalyzed esterification reaction
Titikan Somboon a, *, Ratchanee Phatchana a, Wimonrat Tongpoothorn a, Sira Sansuk b, **
a
Department of Chemistry, Faculty of Engineering, Rajamangala University of Technology, Isan, Khon Kaen Campus, Khon Kaen, 40000, Thailand
b
Materials Chemistry Research Center and Center of Excellence for Innovation in Chemistry, Department of Chemistry, Faculty of Science, Khon Kaen University, Khon
Kaen, 40002, Thailand

A R T I C L E I N F O A B S T R A C T

Keywords: A simple and green procedure for quantifying the ethanol concentration based on the conductometric method
Ethanol and the uncatalyzed esterification reaction was demonstrated. The reaction progress between ethanol and acetic
Liquor samples acid was followed by detecting the electrical conductivity, corresponding to the amount of hydrogen ion and
Conductivity
acetate anion from acetic acid dissociation. By fixing the concentration of acetic acid, the decrease of conduc­
Esterification reaction
tivity with increasing the ethanol concentration was observed. The inversely linear relation between the con­
ductivity and ethanol concentrations in the range of 0.0–99.9%v/v was successfully obtained. The ethanol
content can be efficiently quantified, with good intra-day and inter-day precision (<3.45% RSD). The proposed
method was successfully employed for determining the ethanol concentration in liquor samples, validated by the
GC method. Therefore, this proposed strategy provides an alternative practical method through conductometry
for the quantitative detection of non-electrolytic ethanol solution.

1. Introduction MS) (Stupak et al., 2017), 1H NMR spectrometry (Zuriarrain et al.,

Esterification is a chemical reaction between carboxylic acid and
alcohol producing an ester and water. It is a reversible process and the
reaction progress is extremely slow in the condition without catalyst
(Beula & Sai, 2014). Generally, strong acid catalyst, such as sulfuric acid
or hydrochloric acid can be used to accelerate the reaction. However,
several draw-backs were found in such reaction, for example the dehy­
dration of alcohol or isomerization. Moreover, employment of such
strong acid catalyst may cause corrosion and acidic hazardous waste
(Lynnette & Joan, 2001). It is well known that most alcoholic beverages
contain ethanol and water as the major component. Not only ethanol
and water, the minor composition consists of volatile, e.g., carboxylic
acids and nonvolatile flavour compounds e.g. unfermented sugars and
color substances. Ethyl acetate in beverages can be produced by acetic
acid bacteria in fermentation process and also by esterification of
ethanol and acetic acid over time (Dragone et al., 2009).
In general, the content of ethanol in alcoholic beverages is evaluated
in order to assure the quality and authenticity of drink products.
Recently, the ethanol content in alcoholic drinks has been quantified by
various techniques such as gas chromatography-mass spectrometry (GC-
2015), thermal infrared enthalpimetry (Oliveira et al., 2017, 2018),
Near-Infrared Spectrophotometry (Debebe et al., 2017), electrochemical
enzyme biosensors (Barsan et al., 2016), membraneless gas-liquid sep­
aration flow system (Kraikaew et al., 2019), reversible solid sensor
(Martínez et al., 2018), digital image-based method (Dos Santos Bene­
detti et al., 2015), flatbed scanner and automated digital image analysis
(Curbani et al., 2020), propagating waves (Somboon & Sansuk, 2018)
and colorimetric oscillating signals (COS) (Sansuk et al., 2020). How­
ever, there have been only a few reports on the use of electrochemical
techniques such as amperometry (Paixão et al., 2002) and cyclic vol­
tammetry (Yu et al., 1997) for the determination of ethanol in alcoholic
drinks.
Electrochemical methods have been found to provide a number of
interesting applications in the areas of environment and industry. The
significant electroanalytical techniques include potentiometry, voltam­
metry, amperometry, coulometry and conductometry (Bard & Faulkner,
2000). Typically, almost all methods are based on electrochemical re­
actions except conductometry. Conductivity measures the electrical
current passing through an electrolytic solution and is directly related to
total ion concentrations. The decrease of ion concentrations in the

* Corresponding author.
** Corresponding author.
E-mail addresses: titikan.so@rmuti.ac.th (T. Somboon), sirisan@kku.ac.th (S. Sansuk).

https://doi.org/10.1016/j.lwt.2021.112593
Received 5 July 2021; Received in revised form 20 September 2021; Accepted 6 October 2021
Available online 11 October 2021
0023-6438/© 2021 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
T. Somboon et al. LWT 154 (2022) 112593

solution would be lower the electrical conductivity. In quantitative

analysis, conductometric titration is a simple example (Lubert &
Kalcher, 2010). This method has been successfully applied for the
analysis of various analytes such as acids and bases (Coelho & Gutz,
2006), sulfate (Garcia & Schultz, 2016), cellulose nanocrystal sulfate
half-ester content (Beck et al., 2015), and total volatile basic nitrogen
(TVB-N) (Xia et al., 2016). Also, the electrical conductivity measure­
ments were successfully applied to evaluate the effects of soil variability
on crop growth (Stadler et al., 2015) and to predict the hydraulic con­
ductivity of unsaturated soils (Niu et al., 2015). However, these methods
are limited due to their non-selective characteristic and undetectable
non-electrolytic samples, i.e., ethanol (James, 2004). Therefore, devel­
opment of the experimental strategy based on a simple conductivity
measurement, which is a fundamental technique in general laboratory,
becomes interesting, since this method is considered as a fast, sensitive,
inexpensive and reliable detection.
To the best of our knowledge, the application of conductometry in
quantitative analysis has been less reported, especially its performance
as the analytical tool on the detection of non-electrolytic samples. In this
work, a simple and green strategy for the quantification of non-
electrolytic ethanol in liquor samples based on the conductivity mea­
surement and the uncatalyzed esterification reaction was demonstrated.
The proposed strategy requires two important parts: (i) the specific re­
action of ethanol with a suitable electrolytic reagent was carried out and
(ii) all products from the reaction would be non-electrolytic to avoid the
effect of interfering ions in the detection. In this work, ethanol was
chosen as a representative of non-electrolytic sample, while a weak
electrolytic acetic acid was used. The uncatalyzed esterification reaction Fig. 1. Schematic presentation of the conductometric assay for ethanol
between ethanol and acetic acid was employed to avoid strong acidic determination.
waste. It is noted that ethyl acetate produced in this reaction would be
considered as non-electrolytic compound. By this procedure, the change
5 min dissolved in CDCl3 was investigated by using 1H NMR spectrom­
of conductivity would mostly come from hydrogen ion (H+) and acetate
eter (400 MHz, Varian, UK) to confirm the reaction progress.
anion (CH3COO− ) generated from a dissociation of acetic acid. The
linear relation between conductivity and ethanol concentration was
successfully established. The presented procedure was tested for ethanol 2.2. Calibration and precision of ethanol detection
determination in distilled alcoholic drink samples (whisky and vodka),
also validated with gas chromatography (GC). The conductivity of the studied solution was recorded over time by
using a conductivity meter. To investigate the effect of acetic acid and
2. Materials and methods ethanol concentrations on the conductivity change, the employed con­
centrations of acetic acid were 0.05–30.0 %v/v, while the ethanol
2.1. Materials and instrumental setup concentrations were 0.00–99.9 %v/v. The volume of each solution and
the measurement of electrical conductivity were performed as described
Glacial acetic acid (CH3COOH) was purchased from Merck (Ger­ in Section 2.1. Then, the calibration curve plotted between the con­
many). Ethanol (CH3CH2OH, 99.9%(v/v)) was obtained from RCI Lab­ ductivity of the reacted solution and different concentrations of the
scan (Thailand). Deionized water with conductivity less than 10 μS/cm standard ethanol solution was established. All experiments were carried
from RCI Labscan (Thailand) was used as solvent throughout. The stock out in triplicate at the temperature of 25.0 ± 0.5 ◦ C at a constant stirring
solution of 20%(v/v) CH3COOH was prepared daily with deionized rate of 300 rpm. The linear regression was performed to evaluate the
water. The experimental setup consisted of a jacketed glass beaker (50 linear range and regression coefficient (R2). The precision in terms of the
cm3) connected to a thermostat (Vision scientific VS-310FW, Korea) as intra-day (n = 3) and inter-day (over three consecutive days, n = 3 × 3)
presented in Fig. 1. Water from the thermostat was circulated in and out was also investigated at three levels of ethanol concentration (20, 40 and
of this beaker. Although the uncatalyzed esterification reaction between 60 %v/v). The electrical conductivity (κavg) of each measurement was
acetic acid and ethanol was very slow (Beula & Sai, 2014), the mea­ used to calculate the relative standard deviation (%RSD).
surements were still carried out at room temperature (25.0 ± 0.5 ◦ C) at a
stirring rate of 300 rpm to minimize the energy consumption. The 2.3. Determination of ethanol concentration in alcoholic samples
mixture was conducted in a jacketed glass beaker, in which the fixed
volumes of acetic acid (20 cm3), deionized water (10 cm3) and ethanol In this study, the ethanol content in eight commercial brands of
(10 cm3) were pipetted and mixed inside this unit. Simultaneously, distilled alcoholic samples (whisky and vodka) was quantified by using
electrical conductivity (κ, μS/cm) of the homogenized mixture was the presented method. The concentration of ethanol in all samples was
measured as a function of time by using a conductivity probe connected analyzed without the sample dilution. Ten cm3 of each sample was used
to a conductivity meter (Eutech PC 700, Singapore). The electrical (instead of the standard ethanol solution) and the procedure for the
conductivity at a certain time was employed as a key parameter in this ethanol determination was similar, as described in Section 2.1. The
study. Also, the conductivity at 300 s was chosen for further ethanol average value of solution conductivity (κavg) was employed as a key
analysis. In all experiments, the measurements were carried out in parameter to access the ethanol content in the samples.
triplicate. The mean and standard deviation (SD) of the experimental In addition, the ethanol content in all distilled alcoholic drinks was
data were analyzed. The uncatalyzed esterification reaction between determined by using gas chromatography (GC-450 series) to validate the
20%v/v acetic acid and 20 %v/v ethanol (molar ratio of about 2.04) for proposed method. In this study, the internal standard method was used

2
T. Somboon et al.
to quantify the ethanol concentration and isopropanol was employed as
the internal standard. The working ethanol solutions in the range of
1.0–10.0 %v/v were mixed with 1 cm3 of isopropanol before injecting
into the GC system. The peak area ratio of ethanol to isopropanol at each
concentration level was plotted to construct a calibration curve. Then,
all samples were diluted to 5.0 %v/v and 2 cm3 of each diluted sample
was mixed with isopropanol prior to the GC analysis. The ratio of the
peak area and dilution factor of each sample was taken into consider­
ation for the ethanol content (Sansuk et al., 2020; Somboon & Sansuk,
2018). The statistical analysis of the t-test calculation was performed in
order to compare the efficiency of the presented method with the GC
methods. In this analysis, the t value (n = 7) was considered at a 95%
confidence level to assure the results obtained by these two methods.
3. Results and discussion
3.1. The uncatalyzed esterification reaction between acetic acid and
ethanol
Generally, the esterification between carboxylic acid and alcohol
could be promoted under the presence of an acid catalyst, i.e., H2SO4
(Beula & Sai, 2014). However, such a catalyst was a strong electrolyte
and could interfere with the conductivity measurement. Moreover, the
reaction was carried out without such strong acid catalyst to avoid the
corrosion and production of acidic hazardous waste (Lynnette & Joan,
2001). However, it is well known that the uncatalyzed esterification
reaction has a high activation energy and requires long time to reach an
equilibrium. As presented in Fig. 2, the 1H NMR spectra of the mixture in
CDCl3 at room temperature revealed a signal of acetic acid at δ 2.06 (3H,
s) and three signals of ethanol at δ 5.00 (OH, s), 3.60 (2H, q) and 1.16
(3H, t). Importantly, three crucial signals at 4.10 (2H, q), 2.06 (3H, s)
and 1.22 (3H, t) indicated the production of ethyl acetate, but it was
produced in relatively low amount due to high activation energy and
long time to reach an equilibrium as mentioned. However, the 1NMR
result in Fig. 2 confirmed that the uncatalyzed esterification reaction
between acetic acid and ethanol can occur spontaneously. The use of the
excess amount of acetic acid in such reaction can increase the rate of
esterification because the reaction can be catalyzed by acetic acid itself
(Calvar et at, 2007). The molar ratio of acetic acid to ethanol is the key
parameter to increase the rate of reaction, in which rate of esterification
increases with increasing the molar ratio. In this study, the molar ratio of
Fig. 2. 1H NMR spectrum of the reaction mixture between 20%v/v acetic acid and 20%v/v ethanol in CDCl3.
3
LWT 154 (2022) 112593
acetic acid and ethanol in the condition presented in Fig. 2 is 2.04, which
is relatively high. Therefore, the esterification reaction can be pro­
gressed. As presented in Fig. 2, NMR peak of ethyl acetate is quite small,
indicating that the reaction does not happen quantitatively. However,
the decreased amount of acetic acid under the studied condition may
strongly affect the change of electrical conductivity. Therefore, the re­
action between ethanol and acetic acid in various concentrations on the
electrical conductivity would be further studied.
3.2. Effect of ethanol concentration on the electrical conductivity
The effect of ethanol concentration on the conductivity of the
mixture was investigated. In this experiment, the concentration of acetic
acid was fixed at 20 %v/v. Then, the esterification reaction was carried
out by mixing with different concentrations of ethanol (0–50 %v/v), as
described in Section 2.1. The conductivity was immediately measured
every second after mixing acetic acid, water, and ethanol solution. This
can result in the dilution effect, which ultimately influences the disso­
ciation of acetic acid and consequently the concentration of hydrogen
ion (H+) and acetate anion (CH3COO− ) in solution. The experimental
result in Fig. 3 presents the plot of the conductivity (μS/cm) of the re­
action mixture as a function of time. For all experiments, it was found
that the conductivity suddenly decreased (0–40 s) due to the dilution
effect. Under the condition without addition of ethanol (0 %v/v), the
conductivity is quite constant for entire detection time. For the condition
with addition of ethanol, the slight decrease of electrical conductivity
with time can be observed in all experimental conditions (40–300 s). It is
because the uncatalyzed esterification reaction is extremely slow (Beula
& Sai, 2014). Moreover, the experimental results present that the con­
ductivity decreases with increasing ethanol concentration from 10 to
50%v/v. This results indicate the decreased amount of hydrogen ion
(H+) and acetate anion (CH3COO− ) due to the higher progress in
esterification of acetic acid with ethanol. Considering the molar ratios of
acetic acid and ethanol in each condition in Fig. 3(b–d), the ratios are
4.089, 1.361 and 0.816 for 10%v/v (b), 30%v/v (c) and 50%v/v (d),
respectively. The molar ratio at the condition of 50 %v/v ethanol is
lower than other conditions, resulting in the decreased amount of acetic
acid (low amount of H+ and CH3COO− left in the solution). In this work,
the conductivity at 300 s was chosen for further ethanol analysis.
The behavior of weak electrolytic solution (acetic acid) under the
studied esterification reaction was investigated. The influence of acetic
T. Somboon et al.
Fig. 3. Electrical conductivity as a function of time under the esterification reaction with various ethanol concentrations: (a) without ethanol, (b) 10 %v/v, (c) 30 %
v/v, and (d) 50 %v/v. The concentration of acetic acid was fixed at 20 %v/v.
acid concentrations with fixed ethanol concentration at 20%v/v on the
conductivity was presented in Fig. 4. Each data point represents the
conductivity at 300 s of different acetic acid concentrations (0.05–30 %
v/v). The results show that the conductivity sharply increases at a low
level of acetic acid concentration (0.05–1.0 %v/v), and then slightly
increases in the range of 1.0–30 %v/v. Then, the molar conductivity as a
function of acetic acid concentration in the unit of mole/L was also
investigated and presented in the inset of Fig. 4. The exponential
decrease of molar conductivity with increasing the square root of the
acetic acid concentration was obtained, corresponding to the theoretical
tendency of weak electrolyte (Girault, 2004, p. 143). Therefore, the
results confirmed that the behavior of the weak electrolyte, i.e., acetic
acid still remained under such reaction. This is the advantage of using
the uncatalyzed esterification reaction, in which no strong acid catalyst
(strong electrolytic solution), e.g. H2SO4 or HCl could interfere the
conductivity measurement and no acidic waste is produced after the
experiment (Lynnette & Joan, 2001).
Fig. 4. Conductivity of mixture between fixed ethanol concentration of 20 %v/v and various concentrations of acetic acid. The inset presents the relation between
molar conductivity and the square root of the acetic acid concentration.
4
LWT 154 (2022) 112593
3.3. Calibration for ethanol determination
The influence of the ethanol concentration (0.0–99.9%v/v) on the
conductivity under the esterification reaction with different concentra­
tions of acetic acid (0.05–30.0 %v/v) was studied. Fig. 5 presents the
linear decreases of electrical conductivity in the range of 0.0–99.9%v/v
ethanol concentration with R2 of more than 0.98 in all conditions of
acetic acid As mentioned before, the molar ratio between acetic acid and
ethanol would be decreased, whenever the ethanol concentration was
increased. Therefore, the decreasing tendency in the conductivity in all
conditions presented in Fig. 5(a–h) results from the decreased amount of
H+ and CH3COO− from acetic acid dissociation.
The experimental parameters including the sensitivity (slope of each
calibration plot) and linearity (the value of R2) characterized from the
obtained each linear calibration are shown in Table 1. Considering each
concentration of acetic acid, the slope of linear calibration increased,
indicating that the sensitivity of ethanol detection increased with
T. Somboon et al. LWT 154 (2022) 112593

Fig. 5. The linear relation between the conductivity and ethanol concentration (0–99.9 %v/v) under different concentrations of acetic acid: (a) 30, (b) 20, (c) 10, (d)
5, (e) 1, (f) 0.5, (g) 0.1, and (h) 0.05 %v/v.

intra-day and inter-day precision were less than 2.16% and 3.45%,
Table 1
respectively. These results confirmed a high repeatability of the pro­
Experimental parameters obtained from the calibration plot.
posed method for the ethanol quantification.
[CH3COOH] (%v/v) Linear equation Correlation coefficient (R2) The analytical performance for quantifying ethanol content was also
0.05 y= − 0.6634x + 106.50 0.9954 evaluated in term of the limits of detection (LOD) and quantification
0.10 y= − 0.8248x + 126.41 0.9842 (LOQ). The experiment was performed for seven replicates with 1.0%v/
0.50 y= 1.8652x + 312.42 0.9966
−
v ethanol concentration. The LOD and LOQ were calculated based on
1.00 y= − 2.9342x + 476.41 0.9918
5.00 y= − 6.1914x + 995.33 0.9910 3SD and 10SD, respectively, where SD is the standard deviation of the
10.00 y= − 8.0527x + 1337.10 0.9947 calculated concentration of ethanol (Sansuk et al., 2020). The calculated
20.00 y= − 10.0303x + 1624.69 0.9973 value of SD was 0.21, and then the obtained LOD and LOQ were 0.63 and
30.00 y= − 11.3790x + 1826.30 0.9872 2.10%v/v, respectively. Considering the ethanol content of alcoholic
drinks commercially available in Thailand, the range of ethanol con­
increasing the amount of acetic acid. At 30%v/v acetic acid, the highest centration is about 5.0–40.0 %v/v. The LOD obtained from the proposed
sensitivity was achieved. However, the regression coefficient at this method was much lower than the concentration specified in commercial
condition (R2 = 0.9872) was lower than that at 20%v/v acetic acid (R2 alcoholic drinks. Therefore, the method had a potential to be employed
= 0.9973). Therefore, the obtained calibration at 20%v/v acetic acid as an alternative strategy for the determination of ethanol.
(Fig. 5(b)) was chosen. At this condition, the obtained linear relation
was y = − 10.0303x + 1624.69, where y is the electrical conductivity in 3.5. Analysis of distilled alcoholic samples
μS/cm and x is the concentration of ethanol in %v/v. This linear rela­
tionship would be employed for the determination of the ethanol con­ The method was applied to quantify the ethanol content in eight
tent in real alcoholic drinks. alcoholic drinks including whisky and vodka. In this work, the distilled
samples were chosen in order to avoid the interference from the sample
since beer or wine usually contains other electrolytes, for example,
3.4. Precision and detection limit of the presented method
tartaric and malic acids in wine, or carbonic acid in beer (Mato et al.,
2005). The ethanol content in all samples was analyzed without sample
To validate the presented procedure, the intra-day and inter-day
dilution. The experimental procedure was similar as described in Section
experiments were performed at three levels of ethanol concentration
2.3. By using the obtained electrical conductivity and calibration (Sec­
(20, 40 and 60 %v/v). The intra-day precision was done in triplicate (n
tion 3.2), the ethanol concentrations in whisky and vodka samples were
= 3) in a day, while the inter-day precision was tested over three
successfully determined and summarized in Table 3. The ethanol con­
consecutive days (n = 3 × 3). As presented in Table 2, the %RSDs of
tents were found as follows; 38.44 %v/v for whisky 1, 39.55 %v/v for
whisky 2, 34.05 %v/v for whisky 3, 34.35 %v/v for whisky 4, 38.52 %
Table 2 v/v for vodka 1, 41.34 %v/v for vodka 2, 38.34 %v/v for vodka 3, and
Precision of the proposed method for the ethanol determination. 40.81 %v/v for vodka 4, with the percentage errors in the range of
[EtOH]std Intra-day (n = 3) Inter-day (n = 3 × 3) − 4.07 to 4.15, when compared to the concentrations labeled on the
(%v/v) κavg (μS/cm) a
SD %RSD κavg (μS/cm)a SD %RSD sample bottles.
20 1447.33 17.90 1.24 1456.11 30.83 2.12
40 1195.67 24.19 2.02 1230.00 30.02 2.44 3.6. Validation of the presented method
60 1017.67 21.96 2.16 1020.56 35.25 3.45
a
average conductivity of solution. In order to validate the proposed method, the ethanol concentration

5
T. Somboon et al. LWT 154 (2022) 112593

Table 3
Performance of the proposed method for the ethanol analysis in commercial alcoholic drinks compared with the GC technique.
Alcoholic [EtOH]label (%v/ The proposed method The GC technique
drinks v) a
κsolution (μS/cm) [EtOH]sam % Ratio of peak Dilution [EtOH]dil (%v/ [EtOH]sam (%v/ %
(%v/v) error area factor v) v) error

Whisky 1 40.0 1239.10 ± 38.44 3.89 1.558 ± 0.003 8.00 4.88 39.03 2.41
17.23
Whisky 2 38.0 1228.03 ± 39.55 − 4.07 1.576 ± 0.062 7.59 4.95 37.59 1.07
15.24
Whisky 3 35.0 1283.12 ± 34.05 2.70 1.586 ± 0.011 7.00 4.99 34.94 0.16
19.21
Whisky 4 35.0 1280.15 ± 34.35 1.86 1.584 ± 0.038 7.00 4.98 34.87 0.36
24.51
Vodka 1 40.0 1238.35 ± 38.52 3.71 1.613 ± 0.005 8.00 5.10 40.82 − 2.06
21.63
Vodka 2 40.0 1210.04 ± 41.34 − 3.35 1.552 ± 0.009 8.00 4.85 38.82 2.94
14.12
Vodka 3 40.0 1240.13 ± 38.34 4.15 1.549 ± 0.014 8.00 4.84 38.73 3.17
18.97
Vodka 4 40.0 1215.31 ± 40.81 − 2.04 1.591 ± 0.035 8.00 5.01 40.10 − 0.25
23.56
a
Refers to the average conductivity of the sample solution (n = 3).

in all samples was also determined by GC-FID (Sansuk et al., 2020;
Somboon & Sansuk, 2018). The working ethanol standard solutions in
the range of 1.0–10.0 %v/v were prepared and mixed with isopropanol
internal standard as described in Section 2.3. The ratio of the peak area
of standard ethanol to internal standard (isopropanol) was linearly
increased with ethanol concentrations. The linear equation was
expressed as follows: y = 0.2458x – 0.3590 with R2 of 0.9975, where x
and y are ethanol concentration and the ratio of peak area, respectively.
As mentioned in the experimental section, both whisky and vodka
samples were diluted to 5.0 %v/v before analysis with GC. Thus, the
dilution factor was considered for ethanol determination. By using the
dilution factor and the obtained calibration, the ethanol concentration
in all samples was determined and summarized in Table 2. The results
showed that the percentage error was in the range of − 2.06 – 2.94%,
when compared to the labeled contents on the liquor bottles.
To further validate the performance of the presented method for
ethanol determination in practical application, experiments with spiked
samples were performed. Whisky 4 and vodka 4 were chosen as repre­
sentative samples. Both samples were spiked with 5, 10 and 20 %v/v
ethanol and then were analyzed by using the calibration obtained from
the proposed method and GC. The experimental results as summarized
in Table 4 showed that the recovery obtained by the proposed method
was 92.4–116.7, and the %RSD was 1.52–3.39. The GC results presented
that the recovery was 98.5–107.6, and the %RSD was 0.92–2.85. These
results suggested that the analysis of ethanol by the proposed method
had little effect from the sample interferences, but the precision of
ethanol detection was lower, compared to the GC technique. However,
the advantage of the proposed method was the performance without
sample dilution that can reduce the experimental step. Validated with
the GC, the results confirm that the presented assay can be applied to
determine the ethanol concentration in distilled alcoholic drinks.
Moreover, the statistical analysis of the t-test calculation was per­
formed in order to compare both methods as presented in ESI 1.
Considering a 95% confidence level, the calculation results showed that
the calculated t value was 0.11, which was lower than the critical value
(2.36; n = 7) (Skoog et al., 2004). This result confirmed that there were
no significant differences between the proposed method and the GC
technique. Hence, the presented approach based on the conductivity
measurement in the esterification of acetic acid and ethanol provided a
high capability for the ethanol detection in distilled alcoholic drinks.
4. Conclusion
In summary, the conductometry approach was successfully applied
for the quantitative analysis of a non-electrolytic ethanol solution. The
presented strategy was based on the measurement of electrical con­
ductivity of H+ and CH3COO− generated from the dissociation of acetic
acid left in the uncatalyzed esterification reaction. The conductivity was
employed as a key parameter to access the ethanol content in two types
of distilled alcoholic samples (whisky and vodka). The calibration be­
tween the electrical conductivity and ethanol concentrations was suc­
cessfully employed to quantify the ethanol content in real samples. This
proposed method provided simplicity, reliability, and cost-effectiveness.
Moreover, it could be considered as a green detection method, since the
uncatalyzed esterification reaction produces less strong acid waste.
Therefore, the method could be employed as an alternative method for
the determination of ethanol in real alcoholic samples.

Table 4
Recoveries of ethanol determination in spiked samples obtained from the presented method.
Presented method The GC technique

Alcoholic Sample Spike (%v/v) found (%v/v) Detected (%v/v) Recoverya (%, n = 3) RSD found detected Recoverya (%, n = 3) RSD

(%, n = 3) (%v/ (%v/v) (%, n = 3)

v)

5 34.35 40.18 ± 0.78 116.7 1.95 34.87 40.42 ± 1.06 107.6 2.65
Whisky 4 10 34.35 45.52 ± 0.72 111.7 1.59 34.87 45.06 ± 1.28 101.9 2.85
20 34.35 55.01 ± 1.27 103.3 2.31 34.87 54.96 ± 0.51 100.5 0.92
5 40.81 46.31 ± 0.70 103.5 1.52 40.81 46.07 ± 1.09 105.2 2.36
Vodka 4 10 40.81 50.30 ± 1.70 94.9 3.39 40.81 51.31 ± 1.01 105.0 1.96
20 80.81 59.29 ± 1.28 92.4 2.16 40.81 60.51 ± 1.37 98.5 2.26
a
Recovery (%) = (Amount detected – Amount found) × 100/Spiked amount.

6
T. Somboon et al.
Declaration of competing interest
The authors declare that there is no conflict of interest. This article
does not contain any studies with human or animal subjects.
Acknowledgements
This research project is supported by Rajamangala University of
Technology Isan. Contract No. ENG 30/63. It is also partially supported
by Thailand Science Research and Innovation (TSRI). Contract No.
FRB630010/0174-P6-07. The authors thank Hathaikan Dawangpa and
Primprapa Nasuho for their helps in the sample preparation for the GC
analysis.
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