Chemical Engineering and Processing 54 (2012) 3741

Contents lists available at SciVerse ScienceDirect

Chemical Engineering and Processing:

Process Intensication
journal homepage: www.elsevier.com/locate/cep

Extraction of glycyrrhizic acid from licorice root using ultrasound: Process

intensication studies
Trupti W. Charpe, Virendra K. Rathod
Chemical Engineering Department, Institute of Chemical Technology, Matunga, Mumbai 400019, India

a r t i c l e

i n f o

Article history:
Received 13 August 2011
Received in revised form 11 January 2012
Accepted 17 January 2012
Available online 9 February 2012
Keywords:
Licorice
Glycyrrhizic acid
Ultrasound assisted extraction
Batch extraction
Extraction kinetics

a b s t r a c t
This work deals with the intensication of extraction process of glycyrrhizic acid (GA) from licorice using
ultrasound. Various process parameters such as extraction time, solvent to solute ratio, extraction temperature and ultrasound frequency which affect the extraction yield are optimized. The maximum yield
of 36.4 mg of GA/g of licorice is obtained at optimized parameters of 10 min time, 30:1 solvent to solute
ratio, 40 C temperature and 25 kHz frequency. The yield obtained with ultrasound assisted extraction
(UAE) is compared with that of Soxhlet and batch extraction of GA. It is observed that the ultrasound
assisted extraction not only gives higher yield but also reduces the extraction time as compared to conventional method. Temperature has no effect on ultrasound assisted extraction while higher frequency
of ultrasound gives slightly better extraction yield. Further the kinetics of this extraction process is also
investigated based on the second order rate equation available in the literature. Extraction rate constant,
initial extraction rate and equilibrium concentration for different solvent to solute ratio and temperatures are predicted. Model validation is done successfully by plotting experimental and predicted values
of concentration of GA in extract.
2012 Elsevier B.V. All rights reserved.

1. Introduction
Licorice is extensively used in herbal medicines worldwide
due to its anti-inammatory, anti-viral, anti-allergic, anti-oxidant,
and anti-cancerous properties. Traditionally, it is used for the
treatment of peptic ulcers, asthma, pharyngitis, malaria, abdominal pain, insomnia, infections. The major active component in
licorice is glycyrrhizic acid (GA) while other important constituents
include avonoids, isoavonoids, chalcones, coumarins, triterpenoids, sterols, starch, sucrose and glucose, lignins, amino acids,
amines, gums, volatile oils [1]. Besides medicinal usage as an ingredient in pharmaceutical preparations, GA is also used extensively
as a sweetener or functional additives in tobacco, food and confectionery products.
The traditional solvent extraction technique for extraction of
glycyrrhizic acid from licorice suffers from various disadvantages such as large solvent requirement, higher extraction time,
lower yield and higher extraction temperature and thus requires
development of an effective and economical extraction method.
Applications of various novel techniques such as microwave
assisted extraction, microwave assisted micellar extraction and
supercritical CO2 extraction for the extraction of GA from licorice

Corresponding author. Tel.: +91 22 33612020; fax: +91 22 33612010.

E-mail addresses: vk.rathod@ictmumbai.edu.in,
virendrakrathod@gmail.com (V.K. Rathod).
0255-2701/$ see front matter 2012 Elsevier B.V. All rights reserved.
doi:10.1016/j.cep.2012.01.002

root have been already reported in the literature [24]. Ultrasound

assisted extraction (UAE) is also a promising technique for the
extraction of natural ingredients. The main advantages of ultrasound assisted extraction include the reduced extraction time and
reduced solvent consumption [5]. In addition, ultrasound assisted
extraction can be carried out at a lower temperature which can
avoid thermal damage to the extracts [6]. Inspite of many advantages of UAE, extraction of GA using ultrasound has not been
reported in literature. Therefore, in the present work optimization of extraction process using ultrasound and its kinetic model
building have been carried out.
Ultrasound assisted extraction of many other ingredients from
natural sources has been carried out such as linalool from citrus
ower, essential oil from garlic, oil from axseed, lycopene from
tomato, hesperidine and other avonoids from citrus peel and it
is reported that by the application of ultrasound the extraction of
these natural ingredients is enhanced [711]. In ultrasound assisted
extraction, the extraction rate is enhanced due to cavitation. The
cavity developed grows in size and then collapse abruptly with
the release of energy at enormous rate thus increasing the local
temperature and pressure. Therefore, greater penetration of solvent into cellular material occurs and this improves the release of
cell content into bulk medium. Similarly, the physical effects such
as liquid circulation and turbulence produced by cavitation help
in increasing the contact surface area between the solvent and targeted compounds by permitting greater penetration of solvent into
the sample matrix.

38

T.W. Charpe, V.K. Rathod / Chemical Engineering and Processing 54 (2012) 3741

The parameters which affect the extraction process are type of

solvent, extraction time, solvent to solute ratio, temperature and
ultrasonic frequency. Different solvents such as water, methanol,
ethanol, ammonia etc. are used by different investigators for the
extraction of GA. But, universal and environmental friendly solvent
water has a better solubility of GA in it, hence it is used as a solvent
in this work and other parameters are optimized. The main objective of this work is to develop the optimized ultrasound assisted
extraction process for glycyrrhizic acid from licorice root which
will give higher yield at lesser time than conventional methods. The
second objective is to understand the kinetics and to develop the
model for this extraction process under different parameters to predict extraction rate constant, initial extraction rate and equilibrium
concentration.
2. Materials and methods
2.1. Material
Licorice powder is purchased from local market. Water used
as a solvent is freshly prepared de-ionized water from Millipore
Milli-Q 50. Acetonitrile and acetic acid used as solvent for high
pressure liquid chromatography were analytical grade purchased
from Hi Media Ltd., Mumbai, India. Standard i.e., glycyrrhizic acid
(mono-ammonium salt hydrate 70%) was purchased from Aldrich
Chemical Company, USA.
2.2. Apparatus
Ultrasound assisted extraction has been carried out in a dual
frequency ultrasound cleaning bath (Model 6.5l200 H, Dakshin,
India) of internal dimensions 230 mm 150 mm 150 mm and
tank capacity 6.5 L approx, with an ultrasonic power of 200 W and
frequencies of 25 kHz and 40 kHz, equipped with heater and digital temperature controller/indicator. A selector switch is provided
on the panel to select one operating frequency at a time. Power
variation is possible by varying input AC voltage through autotransformer.
2.3. Ultrasound assisted extraction
The extraction is carried out in a glass vessel kept in an ultrasound bath. The clearance of the glass vessel from the bottom of
the bath (where traducers are xed) is kept constant throughout
the experiments. The measured quantity of the licorice powder is
taken in a glass vessel and required amount of water (solvent) is
added to it. In order to avoid the loss of solvent, the glass vessel
is covered with lid before it is kept to ultrasound bath. The ultrasound is introduced till 20 min. Samples are withdrawn at specic
time interval, and then ltered to get clear extract which is then
diluted and analyzed using HPLC. Extraction experiments are carried out to study effect of parameters which affect the extraction
yield such as extraction time, solvent to solute ratio, extraction
temperature and ultrasonic frequency. Solvent to solute ratio is varied from 10:1 to 40:1 keeping all other parameters constant. The
bath temperature is varied from 30 C to 60 C using the heater provided with the ultrasound cleaning bath and all other parameters
are kept constant. Frequency is studied at 2 levels, i.e., 25 kHz and
40 kHz.
2.4. Soxhlet extraction
Extraction is carried out by using a conventional Soxhlet apparatus which consists of distillation ask placed in oil bath, thimble
holder and the condenser. The licorice powder is packed in lter

paper and placed in thimble holder which is lled with the condensed fresh solvent from the distillation ask. The extraction of
GA takes place in the thimble chamber. When the liquid reaches the
overow level, the liquid moves through the siphon and unloads it
back into the distillation ask, carrying extracted solutes into the
bulk liquid. In solvent ask, solute is separated from the solvent
by distillation. Solute is left in the ask and fresh solvent passes
back into the solid bed material. The operation is repeated until
complete extraction is achieved.
2.5. Batch extraction
Batch extraction is carried out in a glass reactor of capacity
150 ml equipped with a six bladed (pitched blade) glass turbine for
agitation. The measured quantity of the licorice powder is taken
in a glass reactor and required amount of water is added to it.
Mixture is agitated for 30 min. Samples are withdrawn after every
5 min interval and ltered. The clear liquid samples are then diluted
and analyzed for GA using HPLC. Different parameters affecting the
extraction such as extraction time, solvent to solute ratio, speed of
agitation and extraction temperature are optimized and the nal
extraction experiment at the optimized conditions is carried out to
compare it with ultrasound assisted extraction.
2.6. Kinetic model
Extraction of many natural ingredients and kinetic model building of these extraction processes have been reported in the
literature [12,13]. Ho et al., 2005 reported that the kinetics of leaching of water soluble compounds from sapwood of tilia based on the
assumption of mechanism of a second order leaching is best suitable for solid-liquid extraction process [14]. The same approach
is been used here to develop a model which can predict extraction
rate constant, initial extraction rate and equilibrium concentration.
The second order kinetics rate equation can be written as:
dCt
= k(Ce Ct )2
dt

(1)

where k is second order rate constant (g of licorice/mg of

GA min), Ce is equilibrium concentration of GA at saturation (mg
of GA/g of licorice), Ct is the concentration of GA in extract at any
time t.
The integrated rate law for the second order extraction can be
obtained by using boundary condition t = 0 to t and Ct = 0 to Ct and
then after rearrangement the nal equation can be written as:
Ct =

t
(1/h) + (t/Ce )

(2)

where
h = kCe2

(3)

Extraction rate constant k (g of licorice/mg of GA min), initial

extraction rate h (mg of GA/g of licorice min) and equilibrium concentration Ce (mg/g of licorice) can be determined experimentally
from a graph of t/Ct vs t.
2.7. Analytical method
The glycyrrhizic acid is analyzed using HPLC. The HPLC instrument is of Thermo Scientic make which consists of solvent
degassing unit spectra system SCM 1000, gradient HPLC pump
spectra system P4000 and spectra system UV 2000 as a UV detector.
The column used is C18 of dimensions 100 4.6 mm. The wavelength is set at 254 nm. The mobile phase is 70% methanol and 30%
acidied water (1% acetic acid) and the ow rate is 1 ml/min.

h (mg of GA/ g of licorice min)

Ce (mg of GA/ g of licorice)

35

mg of GA/g of Licorice

30
25
20
15
10
5

39

90

0.08

80

0.07

70

0.06

60

0.05

50
0.04

40

0.03

30
20

0.02

10

0.01

0
0

0
0

10

15

20

10

25

20
30
40
Solvent to solute ratio (R)
h

Time (min)
Fig. 1. Effect of extraction time on extraction of GA (Solvent to solute ratio 30:1,
temperature 30 C, frequency 25 kHz).

3. Results and discussion

3.1. Kinetic of ultrasound assisted extraction
The extraction kinetics of ultrasound assisted extraction of glycyrrhizic acid from licorice root is studied using water as a solvent.
Various process parameters i.e., solvent to solute ratio, temperature
and ultrasound frequency are varied. The change in the amount
of GA extracted per g of licorice with time is shown in the Fig. 1.
Curve obtained follows the same nature as that of classical extraction from plant material. Two different rates of extraction can be
observed. First is the fast rate obtained initially which is followed
by the gradual increase in the concentration of GA in the extract. It
can be seen from the Fig. 1 that the rate of extraction of glycyrrhizic
acid is higher initially upto 4 min. Thereafter there is very gradual
increase in extraction and after 10 min no signicant increase is
been observed in the extraction of GA.
3.2. Kinetic model
Kinetic model based on second order rate equation available in
literature [14] is used. The graphs of t/Ct and t for different solvent to solute ratio and temperature are plotted and the values of
extraction rate constant k, initial extraction rate h and equilibrium
concentration Ce are obtained from the slope and the intercept of
these graphs. These values are reported in Table 1. Then the kinetic
model for the prediction of extraction yield of GA for different solvent to solute ratios and temperatures is obtained.
Values of h, k and Ce are dependent on solvent to solute ratio
(R) and temperature (T). Therefore the graphs of h, k and Ce are
plotted against R to see the variation of these parameters with
different solvent to solute ratio. It can be seen from Fig. 2 that

k (g of licorice/ mg of GA min)

T.W. Charpe, V.K. Rathod / Chemical Engineering and Processing 54 (2012) 3741

Ce

50

Fig. 2. Effect of solvent to solute ratio on initial extraction rate h, equilibrium concentration Ce , and extraction rate constant k.

the initial extraction rate and rate constant increase logarithmically whereas the equilibrium concentration changes linearly with
solvent to solute ratio.
From the graphs, the variation in h, k and Ce with respect to
solvent to solute ratio is written in the form of equation.
h = 29.61 lnR 28.35

(4)

k = 0.017 lnR + 0.048

(5)

Ce = 0.173R + 26.96

(6)

Therefore, Ct based upon solvent to solute ratio is obtained by

substituting above equations in Eq. (2) as:
Ct =

t
(1/(29.61 lnR 28.35)) + (t/(0.173R + 26.96))

(7)

This Eq. (7) can be used to predict the extraction of GA for different solvent to solute ratio (ml/g) at a given time with extraction
temperature of 25 C and ultrasonic frequency of 25 kHz.
Similar analysis is done for effect of variation of temperature. As
there is no change in the extraction rate constant k with temperature, only initial extraction rate h and equilibrium concentration Ce
are plotted against temperature. Fig. 3 shows that with the change
in the temperature, the initial extraction rate h and equilibrium
concentration Ce changes logarithmically.
Hence, equations for variation of h and Ce with temperature are
written from graphs of h and Ce against temperature (T) as:
h = 8.321 lnT + 16.6

(8)

Ce = 2.678 lnT + 28.92

(9)

Substituting these equations in Eq. (2),

Ct =

t
(1/(8.327 lnT + 16.6)) + (t/(2.678 lnT + 28.92))

(10)

Table 1
Extraction rate constant, initial extraction rate and equilibrium concentration for different process variables.
Process variable

Level

Initial extraction rate, h (mg

of GA/g of licorice min)

Extraction rate constant, k (g

of licorice/mg of GA min)

Equilibrium concentration of
GA, Ce (mg of GA/g of licorice)

Solvent to solute ratio (ml/g)

10:1
20:1
30:1
40:1

38.38
62.47
74.18
78.36

0.046
0.062
0.069
0.069

28.71
30.17
32.62
33.68

Temperature ( C)

30
40
50
60

44.75
47.49
49.44
50.37

0.0311
0.0312
0.0318
0.0318

37.91
39.00
39.41
39.80

40

T.W. Charpe, V.K. Rathod / Chemical Engineering and Processing 54 (2012) 3741

45
40

50

mg of GA/g of licorice

h (mg of GA/ g of licorice min)

Ce (mg of GA/ g of licorice)

55

45
40
35

35
30
25

30C

20

40C

15

50C

10

60C

30
20

30

40
50
Temperature (T)
h

60

70

Model

0
0

10

15

20

Time (min)

Ce

Fig. 3. Effect of temperature on initial extraction rate h and equilibrium concentration Ce .

This Eq. (10) can be used to predict the amount of GA in extract

for different temperature ( C) at a given time with solvent to solute
ratio of 30:1 (ml/g) and ultrasonic frequency of 25 kHz.
3.3. Optimization of process parameters and validation of model
3.3.1. Effect of solvent to solute ratio
The solvent to solute ratio is an important parameter affecting
the extraction. Therefore, this ratio is varied from 10:1 to 40:1 and
results are shown in Fig. 4. It is observed that the amount of GA
extracted per gram of licorice increases with an increase in solvent to solute ratio up to certain level and then it level off for the
higher solvent to solute ratio. This is because the higher solvent to
solute ratio provides the larger concentration gradient which favors
the mass transfer. As there is a marginal difference in amounts of
GA extracted at the ratio of 30:1 and 40:1, 30:1 is taken as optimized ratio and used for all further experiments. Then to validate
the kinetic model, values of concentration of GA in extract at different time for different solvent to solute ratios are calculated from the
predicted model and plotted in the graph. Fig. 4 shows that there is
a good agreement between the experimental and predicted values
of concentration of GA in extract.

Fig. 5. Effect of temperature on extraction of GA (solvent to solute ratio: 30:1,

frequency: 25 kHz).

3.3.2. Effect of temperature

With increase in temperature, the solubility of GA in
water increases and it was expected that the extraction would also
increase. Therefore extraction is carried out at different bath temperatures i.e., 30 C, 40 C, 50 C and 60 C and results are shown
in Fig. 5. But it is observed that there is no signicant change in
mg of GA extracted per g of licorice with increase in temperature.
Extraction of GA increases from 30 C to 40 C but after that up to
60 C only a marginal change is obtained. The solubility of GA in
water increases with increase in temperature, so the extraction also
increases. Vapor pressure has a great inuence on the occurrence
and the intensity of acoustic cavitation. When the temperature is
low bubbles are few but they collapse with relatively high intensity
which enhances the cell disruption. On the other hand, when temperature is high, though more bubbles are created, they collapse
with less intensity. Surface tension also decreases with the increase
of temperature which affects the bubble formation and collapse.
Thus, at higher temperature bubbles collapse with less intensity
and reduce the mass transfer enhancement [7]. Thus, the counterbalance of increase in the extraction due increases solubility and
decreased mass transfer due to less intensive cavitation bubble
collapse at higher temperature results in the marginal change in
extraction of GA. Values of concentration of GA at different time
for different temperatures are calculated from model obtained and
plotted in Fig. 5. It can be seen that there is good agreement between
experimental and predicted values.

35

mg of GA/g of licorice

30
25
10:01
20
20:01
15
30:01
10

40:01

Model

3.3.3. Effect of ultrasound frequency

Extraction of licorice is carried out at two different frequencies i.e., 25 kHz and 40 kHz to investigate its effect on extraction.
Results are shown in Fig. 6 and it is observed that amount of GA
extracted per g of licorice powder at 40 kHz is slightly higher than
that at 25 kHz. The increase in the extraction of GA at higher frequency is because of the enhancement in the mass transfer due
to higher cavitation. Similar increase in extraction at higher frequency upto 45 kHz is also reported for extraction of salvianolic
acid B from Salvia miltiorrhiza root [15]. However, in the present
work increase in the extraction yield at higher frequency is very less
and the frequency of 25 kHz has been considered to be an optimum
frequency.

0
0

10

15

20

Time (min)
Fig. 4. Effect of solvent to solute ratio on extraction of GA (temperature: 30 C,
frequency: 25 kHz).

3.4. Comparison of ultrasound assisted extraction with batch

extraction and Soxhlet extraction
Fig. 7 shows the comparison of amount of GA extracted per
gram of licorice by batch extraction and by ultrasound assisted

T.W. Charpe, V.K. Rathod / Chemical Engineering and Processing 54 (2012) 3741

40

mg of GA/g of licorice

35
30
25
20

25 kHz

15
40 kHz

10
5
0
0

10

15

20

Time (min)
Fig. 6. Effect of frequency on extraction of GA (solvent to solute ratio: 30:1, temperature: 30 C).

frequency are investigated. The yield obtained with ultrasound

assisted extraction is compared with that of Soxhlet and batch
extraction. It is observed that the ultrasound assisted extraction requires lesser extraction time as compared to conventional
method. Extraction is increased with increase in solvent to solute
ratio up to 30:1. It is also observed that there is only marginal
increase in the extraction of GA with increase in temperature
and with increase in the frequency of ultrasound there is slight
increase in extraction yield. In ultrasound assisted extraction
with nal optimized conditions i.e., 30:1 solvent to solute ratio,
40 C temperature and 25 kHz frequency 36.4 mg of GA/g of
licorice is obtained. So it can be concluded that the ultrasound
assisted extraction of glycyrrhizic acid from licorice is an effective way for extraction with advantages of lower time, lower
heating requirement and higher extraction yield. Kinetic model
has been developed for the prediction of amount of GA in the
extract at given time for different solvent to solute ratio as well
as temperature. The model is validated by plotting experimental and predicted values of amount of GA in extract with good
agreement.
References

40
35

mg of GA/g of Licorice

41

30
25
20

UAE

15

Stirred Cel

10
5
0
0

10

15

20

25

Time (min)
Fig. 7. Comparison of ultrasound assisted extraction with batch exaction using
stirred extractor.

extraction. Using ultrasound assisted extraction 37.30 mg of GA/g

of licorice is obtained which is higher than 28.77 mg of GA/g of
licorice obtained by using a stirred extractor. Although, 37.63 mg
of GA/g of licorice powder is obtained using the Soxhlet apparatus,
the time taken for this is very high i.e., 5 h.
4. Conclusions
In the present work, intensication of extraction of glycyrrhizic
acid from licorice root using ultrasound is carried out. Process
parameters which affect the extraction yield like extraction time,
solvent to solute ratio, extraction temperature and ultrasound

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