Separation and Purification Technology 170 (2016) 224–233

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Separation and Purification Technology

journal homepage: www.elsevier.com/locate/seppur

Insights on the influence of microwave irradiation on the extraction

of flavonoids from Terminalia chebula
R. Yedhu Krishnan a,b, M. Neelesh Chandran a, Vellingiri Vadivel a,c, K.S. Rajan a,b,⇑
a
School of Chemical & Biotechnology, SASTRA University, Thanjavur 613401, India
b
Centre for Nanotechnology & Advanced Biomaterials (CeNTAB), SASTRA University, Thanjavur 613401, India
c
Centre for Advanced Research in Indian System of Medicine (CARISM), SASTRA University, Thanjavur 613401, India

a r t i c l e i n f o a b s t r a c t

Article history: The aqueous extract of Terminalia chebula Retz. plays an important role in traditional medicinal formula-
Received 1 April 2016 tions. Experiments on microwave irradiation during aqueous extraction of flavonoids from T. chebula
Received in revised form 20 June 2016 reveal the influence of microwaves on solid-liquid equilibrium, leading to higher equilibrium flavonoid
Accepted 22 June 2016
concentration in the liquid phase at a fixed flavonoid concentration in the solid phase. As a result, micro-
Available online 23 June 2016
wave assisted extraction (MAE) provided 14.17% enhancement in equilibrium yield, compared to that
obtained in conventional extraction at a temperature of 100 °C and a solvent-to-feed ratio of 40 mL/g.
Keywords:
The effective diffusion coefficient at 40 mL/g increased from 0.68
1011 to 1.31
1011 m2/s due to
Terminalia chebula
Flavonoids
microwave irradiation. Our data conclusively indicates that microwave assisted extraction provides
Microwave assisted extraction higher yield, equilibrium concentration and effective diffusion coefficient. The study has also revealed
Solid-liquid equilibrium the possibility of utilization of MAE for short duration extraction with lower solvent and energy con-
Thermodynamics sumption. The extractions were found to be endothermic and spontaneous, with the spontaneity
improved due to microwave irradiation.
Ó 2016 Elsevier B.V. All rights reserved.

1. Introduction by oxidative stress [6]. They also possess significant antioxidant

Medicinal plants occupy an unequivocal position as the most
important source of new drugs. Increased interest remains in the
extraction and separation of bioactive compounds like flavonoids
and other polyphenols from medicinal plants. Terminalia chebula
Retz. (T. chebula) is a medicinal tree from the Combretaceae family
[1], found abundantly throughout India and other parts of South-
east Asia. It is a rich source of bioactive compounds and hence is
utilized in traditional medicinal systems. The dried fruit of T. che-
bula is one of the constituents of the Ayurvedic medicinal formula-
tion Triphala, the other being Terminalia bellerica and Phyllanthus
emblica. T. chebula finds therapeutic applications due to its laxative,
anti-bacterial, cardiotonic, diuretic, hyperlipidemic and anti-
cancer activity [2–4]. The fruits of T. chebula possess significant
anti-fertility efficacy making it a potential herbal contraceptive [5].
Flavonoids and other polyphenolic compounds possess
therapeutic applications against a wide variety of diseases caused
⇑ Corresponding author at: Seshasayee Paper & Boards Chair Professor in
Chemical Engineering, Centre for Nanotechnology & Advanced Biomaterials
(CeNTAB), School of Chemical & Biotechnology, SASTRA University, Thanjavur
613401, India.
E-mail address: ksrajan@chem.sastra.edu (K.S. Rajan).
activity. Flavonoids have the ability to reduce the free radical
steady state concentrations in the human body [7]. They have
the ability to inhibit specific enzymes and to stimulate some hor-
mones and neurotransmitters [8].
Conventional extraction (CE) techniques like soxhlet and other
related techniques have many constraints. They require a longer
extraction time [9]. The prolonged extraction time in CE processes
leads to loss of flavonoids due to oxidation [10]. This may be
averted by making use of microwave-assisted extraction (MAE),
which provides absolute superiority over the conventional extrac-
tion techniques [11,12]. The main processes occurring during
microwave exposure are selective and targeted heating of materi-
als based on their dielectric constants, followed by ionic conduc-
tion and dipole rotation of the molecules [13]. Ionic conduction
is the electrophoretic migration of ions in the presence of electro-
magnetic field [13]. The friction produced as a result of the resis-
tance of the solution to this migration of ions heats the solution.
The alignment and re-alignment of the dipoles with an applied
field is called dipole rotation [14]. The heat is produced as a result
of this forced molecular randomization of the dipoles at a rate of
4.9
109 times per second [15].
Improved methods of extraction from natural sources are
always cardinal as the extracts find significant applications in

http://dx.doi.org/10.1016/j.seppur.2016.06.039
1383-5866/Ó 2016 Elsevier B.V. All rights reserved.
 R. Yedhu Krishnan et al. / Separation and Purification Technology 170 (2016) 224–233
Nomenclature
A surface area, m2
Bi Biot number
Ch concentration of flavonoids in solvent at time h,
mg QE mL1
CL equilibrium concentration of flavonoids in solvent,
mg QE mL1
CS concentration of flavonoids in solid phase at time h,
mg QE/g
CS equilibrium concentration of flavonoids in solid,
mg QE/g
Da activation energy for diffusion, kJ mol1
De effective diffusion coefficient of flavonoids, m2/s
Dp size of particle, m
D0 pre-exponential constant, m2/s
E energy consumption for microwave irradiation, kW h
Ea activation energy for extraction, kJ mol1
h initial extraction rate, mg QE(mL1) min1
k extraction rate constant, (mL) mg QE1 min1
k0 temperature independent factor, (mL) mg QE1 min1
Ke extraction equilibrium constant
perfumery, textiles, pharmaceuticals, cosmetics and food indus-
try. MAE in its current form is a robust method, being a combina-
tion of microwave and traditional solvent extraction method. MAE
was found to curtail extraction time, solvent consumption and
improve the yields of solanesol from potato leaves and stems
[16] and for the extraction of polyphenols from Clinacanthus nutans
Lindau leaves and stems [17]. MAE has also been used for the
extraction of various bioactive compounds like pectin from
Citrullus lanatus fruit rinds [18], alkaloids from Stephania sinica
[19], essential oil from oregano [20], flavonoids from cocoa leaves
[21], etc.
The choice of solvent in MAE hinges on the solubility of com-
pounds chosen for extraction and on the dielectric properties of
the solvent. Most of the reported works on MAE have utilized
ethanol or methanol or their aqueous solutions as the extraction
solvent [22,23]. However, there are only a few reports on the use
of water alone as the solvent for MAE of active principles from
plants, such as those of Yang et al. [24], Wang et al. [25], Shao
et al. [26] and Torun et al. [27] for extraction of compounds from
Geranium sibiricum Linne, Potentilla anserine rhizomes, Perilla
frutescens leaves and sage leaves respectively. The study of MAE
of compounds from T. chebula using water alone as the solvent
assumes importance, due to the fact that the water extract of
Triphala is used in the preparation of Lauha Bhasma, a herbo-
metallic drug [28] belonging to the Ayurvedic system.
Though the advantages of microwave irradiation during extrac-
tion are well realized, certain questions remain unanswered with
respect to the role of microwave in the intensification of extraction.
The present work aims to address the influence of microwave irra-
diation during extraction (microwave assisted extraction) in the
context of solid-liquid equilibrium, identification of appropriate
conditions for enhanced yield at short-duration microwave expo-
sure at the lowest energy consumption and the modulation of acti-
vation energy for diffusion. For this purpose, experiments were
carried out with T. chebula as the solid phase (feed) and water as
the extraction solvent, embracing a broad array of operating condi-
tions. The data on solid-liquid equilibrium under microwave irradi-
ation, lowest energy consumption for short-duration extraction
and the related discussions are the present work’s unique contri-
butions to literature.
225
KT mass transfer coefficient, m s1
R gas constant, J mol1 K1
Rp radius of T. chebula fruit particle, m
S solvent-to-feed ratio, mL/g
T temperature, °C or K
VL volume of solvent, m3
Yh total flavonoids transferred from T. chebula fruit at time
h, mg QE/g
YL total flavonoids transferred from T. chebula fruit at
equilibrium, mg QE/g
Ymax total flavonoids transferred from T. chebula fruit
determined after exhaustive extraction using multiple
solvents, mg QE/g
Greek letters
h time, min or s
c Temperature extraction coefficient
DG° change in Gibbs free energy, kJ mol1
DH° change in enthalpy, kJ mol1
DS° change in entropy, J mol1 K1
2. Experimental
2.1. Materials
Dried fruits of T. chebula were commercially acquired, and Cen-
tre for Advanced Research in Indian System of Medicine (CARISM),
SASTRA University verified the same. The fruits were milled in a
blender and sieved with a rotary sieve shaker to separate particles
of sizes in the range of BSS 200/+300 mesh (mean particle size –
64 lm). For the determination of total flavonoid content, alu-
minium nitrate and potassium acetate were procured from Hi
Media Ltd., India. Flavonoid standard quercetin was acquired from
Sigma-Aldrich Company, USA. DPPH for the test of antioxidant
activity was purchased from SRL Chemicals, India. Solvents such
as chloroform and methanol were purchased from Merck, Germany.
2.2. Extraction procedures
2.2.1. Microwave assisted extraction (MAE)
Microwave irradiation during extraction (Microwave assisted
extraction – MAE) was carried out using a modified domestic
microwave oven (R-219T (S)/(W), SHARP, Japan). The internal
dimensions of 22-L oven were 319 mm (W)
211 mm (H)

336 mm (D). The outer casing of the oven was drilled for accommo-
dating a reflux condenser on to the top and a magnetic stirrer at
the bottom. The reflux condenser averted any solvent loss during
microwave irradiation, and the magnetic stirrer ensured ideal mix-
ing of the solvent and feed. The temperature control was achieved
using a combination of digital temperature indicator and controller
(Microsensors, India) with a ‘J’ type thermocouple. The duration of
on-off cycles was controlled to achieve the desired temperature.
An energy meter (Visiontek, India) was connected to the micro-
wave unit and the magnetic stirrer. Hence, the energy meter read-
ings account for total energy consumption for microwave
irradiation and that for the operation of magnetic stirrer. The
energy meter readings (ET, kW h) were acquired over the entire
period of MAE for all the experimental conditions. The energy con-
sumption for the supply of microwave (E, kW h) was obtained from
the difference between the energy consumption during MAE with
stirring (ET, kW h) and the energy consumption for stirring alone
226 R. Yedhu Krishnan et al. / Separation and Purification Technology 170 (2016) 224–233

(ES, kW h). For this purpose, the energy required for stirring at each
solvent-to-feed ratio was recorded for the duration of extraction,
however, in the absence of microwave irradiation. 5 g of dried
fruits of T. chebula with different volumes of solvent ranging from
25 to 200 mL was exposed to various durations of microwave irra-
diation. The solvent used for MAE of flavonoids from T. chebula was
water. The samples were recovered at various time points. A sche-
matic diagram of the experimental setup is shown in Fig. 1.
2.4. Experimental design for solid-liquid equilibrium studies
Microwave extraction experiments were carried to determine
the solute concentration in solid phase and liquid phase at equilib-
rium. For this purpose, MAE experiments were accomplished
under isothermal conditions with varying mass of feed and the vol-
ume of solvent covering a wide range of solvent-to-feed ratio (3–
50 mL/g). These experiments were also carried out covering a tem-
perature range of 40–100 °C.

2.2.2. Conventional extraction (CE)

2.5. Analytical method
For comparison, the conventional extraction of flavonoids from
T. chebula was performed using a conventional heat-reflux extrac-
2.5.1. Determination of total flavonoid content
tion with temperature control. Also, an exhaustive extraction of
The total flavonoid content in T. chebula dried fruits was deter-
flavonoids from T. chebula was performed for determining the
mined using aluminium nitrate assay [29,30] and expressed as mg
maximum possible flavonoid content that could be extracted from
quercetin equivalents (QE) per millilitre of extract (mg QE mL1).
T. chebula. A 12 h extraction each with chloroform, methanol and
Quercetin standard curve is shown in Fig. 2. Briefly, diluted sample
water was performed in series with the plant material being recov-
extract (500 lL) was treated with 10 % aluminium nitrate (100 lL),
ered after each stage.

2.3. Experimental design for kinetics and thermodynamics

A full factorial design with two factors viz. solvent-to-feed ratio

(5, 10, 20, 40 mL/g with the mass of solid maintained constant at
5 g and temperature (40, 60, 80, 100 °C) were used for the experi-
mental design to study extraction kinetics and thermodynamics. A
total of 24 experimental trials were performed in triplicates. The
energy consumed by the microwave oven at each experimental
condition was recorded. This full factorial study enabled the deter-
mination of (i) kinetic parameters such as diffusion coefficient and
mass transfer coefficient as a function of temperature; (ii) equilib-
rium parameters such as the equilibrium concentration and equi-
librium yield as a function of temperature and solvent-to-feed
ratio (iii) thermodynamic parameters such as change in enthalpy
and change in Gibb’s free energy. Fig. 2. Quercetin standard curve.

Fig. 1. Experimental setup for microwave assisted extraction.

 R. Yedhu Krishnan et al. / Separation and Purification Technology 170 (2016) 224–233
1 M potassium acetate (100 lL) and 4.3 mL of distilled water. This
mix was incubated for 40 min at room temperature, and the absor-
bance of the sample was noted at 415 nm using a UV–visible spec-
trophotometer (Lambda 25, Perkin Elmer, USA). The assays were
performed in triplicates, and the results reported as mean ± stan-
dard error of the mean.
2.5.2. Test for antioxidant activity
The DPPH assay was carried out to study the antioxidant capac-
ity of aqueous extracts of T. chebula [31]. The dried extracts were
dissolved in distilled water to a concentration range of 0.04 mg/
mL to 10 mg/mL. 100 lL extract and 3.9 mL of DPPH solution
(5 mg in 100 mL methanol) were treated together. The absorbance
was measured at 515 nm (Lambda 25, Perkin Elmer, USA) against
distilled water after 30 min incubation in the dark. Similarly, a con-
trol solution was prepared with distilled water instead of extract
solution. The DPPH radical scavenging activity of the extracts
was determined by using the following equation:
DPPH radical scav enging activ ity ð%Þ
 
absorbance of sample
¼ 1
100
absorbance of control
2.6. Scanning electron microscopy (SEM)
A field emission scanning electron microscope (JSM 6701F,
JEOL, Japan) was used to study the effect of microwave irradiation
on the surface morphology of dried T. chebula fruit particles. The
samples were rendered conductive through deposition of a layer
of gold by sputtering before imaging under high vacuum.
2.7. Gas chromatography–mass spectrometry (GC–MS)
The dried extracts were reconstituted in ethanol for GC–MS. The
extracts were analysed in a gas chromatograph with mass selective
detector in the electron ionization mode (70 eV) (Perkin Elmer
Clarus 500) using a capillary column Elite-5 (30 m
250 lm),
helium as carrier gas at a flow rate of 1 mL/min and split ratio of
1:10. The oven temperature programmed at 60 °C initially was
increased at a ramp rate of 6 °C/min to 150 °C and held at this tem-
perature for 2 min. Further, the temperature was increased at a
ramp rate of 4 °C/min to 280 °C and held for 5 min. The comparison
of chemical compositions with the mass spectral library (NIST
2005) endorsed the identification of compounds.
Fig. 3. Variation of flavonoid concentrations in solvent and solid phases at solvent-to-feed ratio of 40 mL/g with respect to time for MAE carried out at 80 °C.
227
3. Results and discussion
3.1. Kinetics of MAE
A second-order model can be used to explain the kinetics of
extraction (flavonoid concentration, Ch vs time, h data), in terms
of extraction rate constant (k) and equilibrium flavonoid concen-
tration in the liquid phase (CL ) as follows [30,32]:
dC h
2
¼ k C L  C h ð1Þ
dh
Solving and subsequently reducing Eq. (1), its final form can be
represented as:
h 1 h
¼ þ ð2Þ
C h h C L
In Eq. (2). ‘h’ is the initial extraction rate. The flavonoid concentra-
tion in the solid phase at any time (CS) and at equilibrium (CS ) can
be calculated from the maximum flavonoid content in the solid
(Ymax) as follows:
C S ¼ Y max  C h  S & C S ¼ Y max  C L  S ð3Þ
The value of Ymax for T. chebula, as determined by exhaustive extrac-
tion is 50.01 mg QE/g.
Fig. 3 shows the variation of flavonoid concentration in solvent
phase and the solid phase with time, for MAE carried out at 80 °C
with a solvent to feed ratio of 40 mL/g. It can be noted from
Fig. 3 that the flavonoid concentration in solvent phase at different
periods of extraction increased. On the other hand, the flavonoid
concentration in the solid phase decreased with time. This obser-
vation may be attributed to the dissolution and diffusion of solute
from the solid to the solvent phase.
3.2. Conditions at equilibria
Solvent-to-feed ratio is an important variable that influences
the extraction of solute from a solid matrix. The equilibrium con-
centration of the solute in the extract (CL ) was obtained from the
extraction kinetics using Eq. (2). The influence of solvent-to-feed
ratio on the equilibrium concentration is shown in Fig. 4, from
which it is evident that the equilibrium concentration decreased
with increase in solvent-to-feed ratio non-linearly. This observa-
tion could be attributed to the increased extract volume at higher
solvent-to-feed ratios.
228 R. Yedhu Krishnan et al. / Separation and Purification Technology 170 (2016) 224–233
Fig. 4. Effect of solvent-to-feed ratio on equilibrium concentration of flavonoids in solvent phase and the concentration of solute remaining unextracted in the solid phase
under equilibrium conditions for MAE at temperature of 80 °C.
In the context of solute recovery from a solid matrix, the influ-
ence of solvent-to-feed ratio is reflected in the concentration of
solute remaining unextracted in the solid phase under equilibrium
conditions. It is evident from Fig. 4 that the concentration of solute
remaining unextracted in the solid phase decreased with increase
in the solvent-to-feed ratio. Lower the concentration of solute
unextracted remaining in the solid phase, higher is the solute
recovery. This indicates that higher solute recoveries are achiev-
able when extraction is carried out at higher solvent-to-feed ratios.
It is to be noted that the solute concentration remaining unex-
tracted did not decrease linearly with increase in the solvent-to-
feed ratio as evident from the rapid reduction in unextracted solute
concentration till the solvent-to-feed ratio of 30.
It is well established that longer contact time between solid and
liquid is likely to result in equilibrium during which the chemical
potential of solute in the solid phase equals that in the liquid
phase. The chemical potentials are directly related to solute con-
centrations. Accordingly, the equilibrium relationship may be
expressed in terms of solute concentrations in the solid phase
and solvent phase respectively. Such a plot is shown in Fig. 5 for
MAE of flavonoids from T. chebula at 80 °C. The data for CE at
80 °C is also shown for comparison. It may be observed from
Fig. 5 that the equilibrium data is non-linear for the range of equi-
librium concentrations investigated here. This observation may be
Fig. 5. Relationship between equilibrium concentration of flavonoids in solvent
phase and solid phase.
interpreted by drawing an analogy with equilibrium data for solute
extraction from a liquid using another immiscible liquid (liquid-
liquid extraction). The equilibrium relationship for liquid-liquid
extraction is linear for dilute solutions, while the same for concen-
trated solutions is non-linear. In the present study, the non-
linearity of equilibrium data for MAE of flavonoid from T. chebula
could be attributed to the relatively higher concentration of solute
in the liquid phase (maximum of 0.5 wt/vol% in the extract). It is
pertinent to mention that equilibrium data for solute recovery
using MAE have not been reported. While comparing the equilib-
rium data for MAE with those of CE, it may be observed that the
equilibrium data in MAE are shifted to the right. In other words,
for the same concentration of solute remaining in the solid phase
at equilibrium, the solute concentration in the extract (liquid
phase) prepared by MAE is higher than that prepared using CE.
For example at the equilibrium solute concentration of 16.05 mg/
g in the solid phase, the corresponding equilibrium concentrations
in CE and MAE were 0.85 mg/mL and 1.15 mg/mL respectively.
It is known that the microwave irradiation improves the kinet-
ics of extraction [33]. However, the capability of microwave expo-
sure to influence the solid-liquid equilibria is not clearly
demonstrated. A plausible explanation for the influence of micro-
wave irradiation on solid-liquid equilibria can be offered by micro-
wave’s capability to unlock solute species hitherto unavailable for
contact with the solvent. To test this hypothesis, conventional
extractions were carried out for 2 h at chosen solvent-to-feed ratio
and temperature, within which solid-liquid equilibria had been
fairly established. This was followed by exposure to microwave
for a short duration. The results of such experiments are shown
in Fig. 6a and b. It is evident from Fig. 6a and b that the flavonoid
concentration in the extract was increased following short dura-
tion exposure to microwave. For instance, the equilibrium concen-
tration in the extract prepared with the solvent-to-feed ratio of
5 mL/g and T = 40 °C by conventional extraction was found to be
4.688 mg QE/g. The flavonoid concentration in the extract was
increased to 5.1819 mg QE/g after microwave exposure (Fig. 6a).
The increase in flavonoid concentration due to microwave expo-
sure post establishment of solid-liquid equilibrium in conventional
extraction was observed for other solvent-to-feed ratios and tem-
peratures also (Fig. 6a and b).
The influence of temperature on the equilibrium yield of flavo-
noids is shown in Fig. 7, from which it is evident that the equilib-
rium flavonoid yield increased with extraction temperature in
 R. Yedhu Krishnan et al. / Separation and Purification Technology 170 (2016) 224–233
Fig. 6. Influence of microwave irradiation on the solid-liquid equilibria (a) at
solvent-to-feed ratio of 5 mL/g (b) at solvent-to-feed ratio of 40 mL/g.
Fig. 7. Effect of temperature on equilibrium yield of flavonoids.
MAE. This could be attributed to the fact that increase in tempera-
ture favours forward reaction, which in this case, is the solute
transfer from solid to liquid phase. The increase in flavonoid yield
is reflected as the proportionate increase in the equilibrium con-
centration of flavonoids also.
The temperature extraction coefficient (c) can be used as a
quantitative measure to represent the increase in flavonoid yield
for every 10 °C rise in temperature [34], as shown in Eq. (4).
Y L ¼ Y L0  c10
T
ð4Þ
229
Fig. 8. Effect of solvent-to-feed ratio on temperature extraction coefficient.
The temperature extraction coefficients have been determined
to be greater than one inline with increased flavonoid yield at
higher temperatures. The temperature extraction coefficients in
MAE seem to be dependent on the solvent-to-feed ratio also
(Fig. 8). Higher temperature extraction coefficients were observed
at higher solvent-to-feed ratios. This suggests that the effect of
temperature on the flavonoid yield is amplified at higher solvent-
to-feed ratios. This observation could be attributed to two effects:
(i) The equilibrium concentrations are normally lower at higher
solvent-to-feed ratios and hence more solute can be further
accommodated at higher solvent-to-feed ratios (ii) The microwave
energy supplied for extraction at a fixed temperature increases
with the increase in the solvent-to-feed ratio and hence the role
of microwave irradiation may be amplified at higher solvent-to-
feed ratios. There is a small difference in temperature extraction
coefficient between the MAE and CE (c = 1.05 for MAE and
c = 1.04 for CE). This small difference could be attributed to the
second effect as discussed above. The higher temperature extrac-
tion coefficients are beneficial when solute extraction from solid
matrix need to be carried out under conditions of limited solvent
supply.
3.3. Influence of energy consumption on cumulative yield
It may be recollected that the flavonoid concentration in the
extract was measured as a function of time (2–120 min). However,
the majority of extraction was completed within about 20–30 min
for all the extraction conditions. The flavonoid concentration at
any time (Ch) was used to determine cumulative yield (Yh), which
is the product of solvent-to-feed ratio (S) and Ch. The terms cumula-
tive yield (Yh) and energy consumption for microwave irradiation (E)
refer to the flavonoid yield obtained and energy consumption
recorded during ‘h’ min of microwave assisted extraction beginning
at h = 0. For a fixed solvent-to-feed ratio, the variation of cumulative
yield (Yh) with energy consumption for microwave irradiation (E) for
different extraction temperatures and extraction times appear to fall
on the same curve (Fig. 9). In essence, it is prudent to conclude that
the solvent-to-feed ratio (S) and energy consumption for microwave
irradiation (E) influence the cumulative yield. Therefore, for a fixed
solvent-to-feed ratio, nearly same cumulative yield is expected for
fixed energy consumption for microwave irradiation (E), irrespec-
tive of extraction temperature and time. The following Yh-S-E rela-
tionship has been developed from the present experimental data,
encompassing cumulative yield range of 25.3896 mg QE/g to
42.2779 mg QE/g, energy consumption for microwave irradiation
230 R. Yedhu Krishnan et al. / Separation and Purification Technology 170 (2016) 224–233

Fig. 9. Variation of cumulative flavonoid yield with energy consumption.

range of 2.981
103 kW h to 3.840
102 kW h and the solvent-
to-feed ratio range of 5–40 mL/g.
Y h ¼ 21:582e7:8019Eh S0:074428
Eq. (5) predicts the present experimental data with R2 of 0.9300,
RSD of 4.82% and the Average absolute relative deviation (AARD)
of 2.26%.
Another noteworthy feature of Yh-E-S relationship for extrac-
tion times of 2–30 min is the stronger dependency of cumulative
yield on energy consumed for microwave energy irradiation (expo-
nential increase) than on the solvent-to-feed ratio (power law
model with a low exponent), with no obvious intermediate ‘E’
and ‘S’ values at which the yield is maximised. The microwave
energy absorbed by the solvent is used for its temperature change
(sensible heat) and phase change (latent heat of vaporisation). At
comparable values of the amount of energy consumed for
microwave irradiation, the temperature rise is higher at lower
solvent-to-feed ratio owing to the lower volume of solvent. Hence,
it is possible that a short duration exposure to microwave irradia-
tion may bring out a large temperature change under conditions of
lower solvent-to-feed ratio. Temperature influences both the
extraction rate constant and the equilibrium concentration and
hence its influence on initial rate are higher. This could result in
ð5Þ higher yield for short duration microwave exposure at lower
solvent-to-feed ratio (S) and lower energy consumption for micro-
wave irradiation (E) than those at higher values of S and E. The
effect of solvent-to-feed ratio (S) and energy consumption for
microwave irradiation (E) on the cumulative yield after 1 min of
MAE is shown as contour plot in Fig. 10. It is evident that similar
or higher cumulative yield can be obtained even at certain lower
solvent-to-feed ratio and lower energy consumption. For instance,
23.7232 mg QE/g of flavonoid yield was obtained in one minute
using the solvent-to-feed ratio of 5 mL/g and energy consumption
of 6.141
103 kW h, while 23.2565 mg QE/g of flavonoid yield
was obtained at the same time using the solvent-to-feed ratio of
10 mL/g and energy consumption of 7.041
103 kW h. The MAE
of 1 min at the solvent-to-feed ratio of 20 mL/g consumed
7.441
103 kW h of energy to produce flavonoid yield of
23.3458 mg QE/g. Hence, short duration microwave assisted
extraction represents an intensified extraction with higher flavo-
noid yields obtained at lower solvent and energy consumption.

Fig. 10. Contour plot for cumulative yield after 1 min of MAE.
 R. Yedhu Krishnan et al. / Separation and Purification Technology 170 (2016) 224–233 231

3.4. Antioxidant activity
The extracts obtained after microwave heating and conven-
tional heating at the temperature of 100 °C and the solvent-to-
feed ratio of 40 mL/g were dried to study the antioxidant activity.
The study also helped in identifying the influence of microwave
irradiation on the antioxidant activity of T. chebula fruits. Fig. 11
shows the % DPPH inhibition at various concentrations. It can be
observed from Fig. 11 that the antioxidant activity of the extracts
prepared by both microwave and conventional routes were analo-
gous. This substantiates the fact that microwave irradiation did not
cause the deterioration of extracts.
3.5. Diffusion coefficient and mass transfer coefficient
Table 1 shows the equations utilized for calculating effective
diffusion coefficient (De), mass transfer coefficient (KT), Biot
number (Bi), activation energy for extraction (Ea), activation energy
for diffusion (Da), extraction equilibrium constant (Ke), change in
enthalpy (DH°), change in entropy (DS°) and change in Gibbs free
energy (DG°).
The diffusion coefficient and mass transfer coefficient for the
MAE and CE are shown in Table 2. It is evident from Table 2 that
the mass transfer coefficients were elevated in MAE in comparison
to that in CE. The Biot number for all extraction experiments (both
MAE and CE) was calculated to be greater than 50. Hence, there
was sufficient mixing between the feed and solvent phases leading
to negligible external mass transfer resistance, and therefore, the
internal transfer was rate-limiting [37].
The activation energy (Ea) for MAE of flavonoids from T. chebula
was found to be 13.46 kJ/mol while that for CE was found to be
13.54 kJ/mol. This shows both the microwave and conventional
extraction processes are governed by diffusion [41].
The reduction in diffusional resistance is evident from higher
effective diffusion coefficient (De) for MAE (De = 0.68
1011 m2/s
at T = 40 °C) when compared to that of CE (De = 0.47
1011 m2/s
at T = 40 °C). The activation energy for diffusion (Da) in MAE
(4.51 kJ/mol) was reduced substantially when compared to the acti-
vation energy in CE (9.87 kJ/mol). The reduction in the activation
energy for diffusion indicates that the diffusion of solute through
the plant (solid) matrix under microwave irradiation is much easier
than that through the plant matrix unexposed to microwave irradi-
ation. It is understood that the higher density of pores and cracks
facilitate solute diffusion. The scanning electron micrographs of
the spent solid obtained after MAE and CE reveal a higher pore den-
sity in the former (Fig. 12), plausibly explaining the ease of diffusion
through plant matrix exposed to microwave irradiation and hence
the reduced activation energy. This confirms that the microwave
irradiation augmented the mass transfer of the solute from solid
to liquid phase.

3.6. Change in Gibb’s free energy

Fig. 11. Comparison of antioxidant activity of aqueous extracts of T. chebula.
Table 3 gives the change in Gibbs free energy, change in
enthalpy and change in entropy for MAE and CE.
Table 1 The higher values of extraction equilibrium constant in the case
Equations used for parameter estimation. of MAE across all temperatures show the positive influence of
Parameter Equation used References microwave irradiation. DH° was observed to be positive indicating
    2   2 the MAE to be endothermic. Positive values of DS° signify the
ep h
Effective diffusion coefficient þ ln p6 [35]
YL
ln Y  Y ¼ DRp
(De) Lh
2
increased disorder due to the mass transfer from solid to solvent
Mass transfer coefficient (KT) C
ln C  C
L
¼ KVTLA h [36] phase. They also indicate the solid-liquid extraction process to be
L h

Biot number Dp K T [37] irreversible. The spontaneity and feasibility of the extraction sys-
Bi ¼
tem were confirmed from the negative values of DG°. The more
De
Ea
Activation energy for extraction k ¼ k0  e RT [38]
(Ea) negative values of DG° at higher temperatures specify the
Activation energy for diffusion De ¼ D0  e
Da
RT [39] increased favourability of the extraction at higher temperatures.
(Da) In comparison to CE, DH° values were higher in MAE indicating
Extraction equilibrium constant Y L [40]
Ke ¼ higher energy absorption and transformation of energy into heat
ðY max Y L Þ
Change in enthalpy (DH°) and ln K e ¼  DRT H
þ DRS

[34] in the presence of microwave irradiation [34]. DS° values were also
change in entropy (DS°) observed to be higher in the case of MAE due to a greater degree of
  
Change in Gibbs free energy DG ¼ DH  T DS [34]
extraction achieved under the influence of microwave irradiation.
(DG°)
The DG° values were more negative for MAE than CE at the same

Table 2
Comparison of effective diffusion coefficient, mass transfer coefficient and Biot number for extraction of flavonoids from T. chebula at solvent-to-feed ratio of 40 mL/g.

T (°C) MAE CE
De
1011 (m2/s) KT
105 (m/s) Bi De
1011 (m2/s) KT
105 (m/s) Bi
40 0.68 ± 0.01 1.13 ± 0.20 123.34 ± 6.08 0.47 ± 0.06 0.89 ± 0.00 120.46 ± 1.02
60 0.80 ± 0.05 2.07 ± 0.29 167.84 ± 9.75 0.54 ± 0.03 1.42 ± 0.00 166.08 ± 3.52
80 1.07 ± 0.08 2.82 ± 0.21 183.97 ± 12.50 0.77 ± 0.05 1.89 ± 0.00 170.98 ± 15.04
100 1.31 ± 0.02 3.38 ± 0.11 230.60 ± 2.16 0.83 ± 0.02 2.74 ± 0.02 210.46 ± 7.96
232 R. Yedhu Krishnan et al. / Separation and Purification Technology 170 (2016) 224–233

Table 4
Chemical compositions of T. chebula aqueous extracts by MAE and CE.

Compounds % Peak area in

extract prepared
using
MAE CE
4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl- 1.3797 0.6909
Benzene, 1,10 -(1-methylethylidene)bis[4-methoxy- 26.7876 20.4099
1,2,3-Benzenetriol 70.7389 78.2529
Other minor compounds 1.0938 0.6463

temperatures showing the positive influence of microwave irradi-

ation in achieving higher spontaneity.

3.7. Chemical composition through GC–MS

The chemical composition of aqueous extracts of T. chebula

fruits obtained by both microwave assisted and conventional
extractions at the solvent-to-feed ratio of 40 mL/g and temperature
of 100 °C was studied using GC–MS (Table 4). The presence of fla-
vonoid fraction 4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-
methyl- was observed in both the extracts. The % peak areas for
the flavonoid fraction in the extract were 1.3797% for MAE and
0.6909% for CE. The major fraction present in both the extracts
was observed to be 1,2,3-Benzenetriol (>70%). The study corrobo-
rates that the chemical composition of the extract was indepen-
dent of the method of extraction.

4. Conclusion

In this work, the influence of microwave irradiation on the

Fig. 12. SEM micrograph images of (a) Raw T. chebula, (b) T. chebula after
microwave assisted extraction and (c) T. chebula after conventional extraction.
extraction of flavonoids from T. chebula has been investigated.
Microwave-assisted extraction (MAE) using the solvent-to-feed
ratio of 20 mL/g yielded 23.35 mg QE/g of flavonoid in one minute
of extraction with the 7.44
103 kW h of energy consumed for
microwave irradiation while 32.72 mg QE/g of flavonoid yield
was obtained at solvent-to-feed ratio of 5 mL/g and energy con-
sumption of 6.14
103 kW h. Hence, the short duration micro-
wave irradiation can be used to achieve a targeted yield using
lower solvent and energy consumption. The thermodynamic
parameters revealed the MAE to be endothermic, irreversible, fea-
sible and spontaneous. The study of activation energy for extrac-
tion indicated that the extraction was governed by diffusion. A
larger number of pores and cracks were formed in the solid phase
after microwave irradiation, as visualised using scanning electron
microscope. The lowering of the activation energy for diffusion
during microwave irradiation is attributed to the formation of
pores and cracks. The effective diffusion coefficient, mass transfer
coefficient and temperature extraction coefficient were higher in
the case of MAE compared to convention extraction (CE). The DPPH
assay revealed that the antioxidant activity of extracts from MAE
and CE were comparable. GC–MS data showed that microwave
irradiation did not contribute to change in the chemical composi-
tion of the extract.

Table 3
Thermodynamic parameters for MAE and CE of flavonoids from T. chebula.

T (K) MAE CE
Ke DH° (kJ/mol) DS° (J/molK) DG° (kJ/mol) Ke DH° (kJ/mol) DS° (J/molK) DG° (kJ/mol)
313 1.71 17.12 58.36 1.14 1.27 11.91 40.01 0.61
333 2.11 2.31 1.65 1.42
353 2.81 3.48 2.10 2.21
373 5.15 4.64 2.67 3.01
 R. Yedhu Krishnan et al. / Separation and Purification Technology 170 (2016) 224–233
Acknowledgements
This work is supported by (i) SRF Grant No.
09/1095/0007/2014-EMR-I, Council of Scientific & Industrial
Research (CSIR), India (ii) Grant No. SR/FST/ETI-331/2013, FIST,
Department of Science & Technology (DST), India (iii) SASTRA
University.
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