Industrial Crops and Products 77 (2015) 920931

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Industrial Crops and Products

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

Optimization of evaporative extraction of natural emulsier cum

surfactant from Sapindus mukorossiCharacterization and cost
analysis
Abhirup Basu, Soham Basu 1 , Sujaya Bandyopadhyay, Ranjana Chowdhury
Chemical Engineering Department, Jadavpur University, Kolkata 700032, India

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

Article history: In the recent world, biosurfactants have gained much research interest because they are eco-friendly
Received 26 June 2015 and non-toxic in nature. The detergency of a naturally available biological surfactant (soap nut) has been
Received in revised form compared with that of a surface-active petroleum waste-derivative (2-hydroxybiphenyl) (2-HBP)1 and
28 September 2015
a widely used commercial surfactant (Ezee). Surface tension (ST),2 critical micelle concentration (CMC)3
Accepted 3 October 2015
and emulsication index (E-24 index)4 are the three principal properties that have been investigated.
Moreover, E-24 index and yield for soap nut have been optimized as a function of extraction time and
Keywords:
temperature by central composite design (CCD)5 in response surface methodology (RSM)6 using Design
Soap nut
Bio-emulsier cum surfactant
Expert Version 8.0.7.1 (StatEase, USA). The optimized emulsication index is observed to be 88.042%
Evaporative extraction at 100 C and 12.02 min, which is higher than that of 2-hydroxybiphenyl (64%) and Ezee (79.6%). The
Optimization maximum achievable yield of the soap nut extract at this temperature and time is 31.21% (g extract/g
Response surface methodology (RSM) of raw pulp of soap nut). The E-24 index of soap nut extract under optimum conditions of evaporative
Cost analysis extraction routeis comparable to the E-24 index of extract from Soxhlet extractor.It has been observed
that optimally extracted soap nut solution has a CMC of 23 kg/m3 , whereas the values of CMC of 2-HBP
and Ezee are 0.2 kg/m3 and 250 kg/m3 respectively. A cost analysis based on the production cost of soap
nut extract and the market price of 2-HBP and Ezee suggests that the price of surfactants varies inversely
with their CMC value and hence, lesser amount of costlier surfactant is required for the reduction of
surface tension in the same extent. The soap nut extract is the best product among the three for the
purpose of emulsication, particularly suitable in food and pharmaceutical industries.
2015 Elsevier B.V. All rights reserved.

1. Introduction consist of two parts a hydrophilic (water-loving) head and a

hydrophobic tail. The hydrophobic part is non-ionic and consists of
Surfactants stabilize emulsions, reduce surface tension and a long hydrocarbon chain while the hydrophilic part is ionic and
are responsible for the detergency action exhibited by soaps and consists of polar groups such as -COO Na+ . When a surfactant
detergents. These are amphiphilic compounds, comprising of both is dissolved in water, the molecules tend to cluster towards the
hydrophilic and hydrophobic moieties and having the ability to surface such that the hydrophobic tails of the surfactant molecules
concentrate on the air water interface. All surfactant molecules are directed away while the polar parts are directed towards water
mass (Muntaha and Khan, 2015). Now, when a piece of fabric
stained with oil/grease is immersed in the surfactant solution fol-
lowed by an adequate degree of agitation, the surfactant molecules
Corresponding author. Fax: +91 033 2414 6378.
tend to aggregate around the oil molecules such that their non-
E-mail addresses: basuabhirup93@gmail.com (A. Basu),
sbasu.juchem@gmail.com (S. Basu), chatterjeesujaya@yahoo.co.in polar hydrophobic part points towards the oil while the polar part
(S. Bandyopadhyay), ranjana.juchem@gmail.com (R. Chowdhury). is directed towards the water. This exerts a net force on the grease
1
Equal contribution to 1st author. or oil due to which they are dislodged and can be washed away
1
2-hydroxybiphenyl 2-HBP.
2
easily.
Surface Tension ST.
3
Critical Micelle Concentration CMC. Synthetic surfactants cause environmental and toxicological
4
Emulsication Index E-24 index. problems as they release carcinogenic toxins in the environment
5
Central composite design CCD. (Reddy et al., 2012). In this respect, natural biosurfactants provide
6
Response surface methodology RSM.

http://dx.doi.org/10.1016/j.indcrop.2015.10.006
0926-6690/ 2015 Elsevier B.V. All rights reserved.
 A. Basu et al. / Industrial Crops and Products 77 (2015) 920931
the added advantage of being less toxic, more biodegradable, higher
foaming capacity, highly selective and specically active at extreme
temperatures, pH and salinity (Samanta et al., 2013).
Soap nuts or reetha (Scientic Name: Sapindus mukorossi) are
obtained from the genus of shrubs Sapindus, commonly referred
to as the Chinese. Sapindus actually comprises of about 512
species of shrubs in the Lychee family, Sapindaceae that consists
of around 2000 species (Azhar et al., 1993). Soap nuts consist of
saponins (Ghagi et al., 2011; Mahar et al., 2011), in the range
of 610% (Kommalapati et al., 1998), which is responsible for
imparting surface active properties of the extract (Chandran et al.,
2013). Saponins are classied as glycosides consisting of nonpo-
lar aglycones called sapogenin and one or more monosaccharides
(Zhou et al., 2013). The main structure of saponin in the peri-
carp shell of reetha fruit is shown in Fig. 1 (Huang et al., 2008).
The strong surface active property of saponins is imparted by the
presence of both polar (sugar) and non-polar (steroid or triter-
pene) groups. The growing demand for saponins in the world
market is attributed to their ecological adaptability (Fiechter, 1992;
Desai and Banat, 1997), their biological and environmental sta-
bility. Soap nut extract nds extensive use in detergents (Cheeke,
2000), soil-washing (Zhou et al., 2013), pharmaceutical industries
(Robbers et al., 1996), enhanced oil recovery (El-Batanoney et al.,
1999; Gong et al., 2005; Chhetri et al., 2009) and environmental
remediation (Urum and Pekdemir, 2004). Moreover, soapnuts have
some medicinal properties such as anti-inammatory (Ankamwar
et al., 2005) and anti-microbial activities (Huang et al., 2007). Their
wide application in Ayurvedic industries is to treat diseases like
eczema, psoriasis, and for removing freckles (Awen et al., 2010).
Although soap nuts have been traditionally in use as a detergent
for decades, no effort has so far been reported on the optimization
of parameters for extraction of surface active saponin rich chem-
ical from the fruit. However, the identication of the optimum
extraction condition is necessary for its successful commercial
application.
In the present study, it has been attempted to nd different sur-
factant properties of soap nut, which is a natural biosurfactant, and
has been compared against those of a biosurfactant, namely, 2-HBP,
produced through biodesulfurization (Kim et al., 2004) of diesel
and a commercial surfactant (Ezee). 2-HBP has been selected for
comparison because it is a biosurfactant or hydrotrope which has
a strong potential to be used as a surface active agent in detergent
industry, and also for enhanced oil recovery (Bandyopadhyay et al.,
2013). It is also expected that with the popularization of biodesul-
furization of diesel producing 2-HBP as a byproduct, this chemical
will also enter the surfactant market in the form of detergent and
will compete with conventional detergents and the bio-detergents
like reetha extract etc.
The properties that have been considered to parameterize sur-
factant activity are surface tension, critical micelle concentration
and emulsication index. The emulsication index of the soap nut
extract is found to be dependent on two independent variables,
i.e. extraction time and extraction temperature and therefore
requires optimization. The conventional methods of optimization
based on one factor at a time require a lot of experiments to be
carried out (Appavoo et al., 2014). Response Surface Methodology
(Yuan et al., 2008; Hasan et al., 2010), a statistical method, is a exi-
ble method used for multi variable optimization (Bidin et al., 2009;
Aziz et al., 2014). Basically the optimization process using CCD in
RSM involves conducting the statistically designed experiments,
tting the experimental data in a mathematical model and check-
ing the adequacy of the model (Sahu et al., 2009). Thus, under the
present study, RSM has been used to optimize extraction time and
temperature for extraction of surface active liquid from soap nut.
The economic viability of the optimally operated extraction process
of soap nut has also been tested through cost analysis against the
921
commercially available counterparts, namely 2-HBP and a deter-
gent, Ezee.
2. Materials and methods
2.1. Materials used
Soapnuts (reetha), the deadly fruits of the Chinese Soap-
berry Tree, produced from agricultural elds were purchased
from local markets. Parafn liquid (LOBA Chemie Pvt., Ltd.), 2-
hydroxy-biphenyl (SigmaAldrich), commercial surfactant, Ezee,
(M/s Godrej Ltd.) were used in the study.
2.2. Evaporative extraction of surfactant from soapnut using
heating mantle arrangement
The evaporative extraction may be described as the leaching
(Treybal, 1980) process in which the solvent, i.e., water, is directly
in contact with the solid (soap nut) from which the solute, i.e. the
saponin is to be extracted with simultaneous concentration of the
extract through the evaporation of water.Since the shell of the fruits
acts as the source of surfactants, soapnuts were crushed in a mortar
and the seeds were removed. 3.5 g of pulp in 100 ml of distilled
water was taken in a round bottom ask and heated in a heating
mantle. The mouth was sealed with cotton to restrict the passage
of vapour and a thermometer was inserted inside the ask. The
temperature was allowed to rise in the ask and after the desired
temperature was achieved, the solution was heated under steady
state for the required time. The step was carried out for different
extraction time and temperature. However, this is not exactly true,
as the temperature controller facilitates alternate cycles of heating,
such that the overall temperature oscillates about a mean value. The
solution was cooled and ltered to remove the remaining pulp from
the solution and the ltrate was analyzed using FTIR spectroscopy
and was tested for the determination of surface tension and the
E-24 index.
2.3. Extraction of surfactant using soxhlet apparatus
The previously used heating mantle arrangement suffered a
huge solvent loss during saponin extraction resulting in the reduc-
tion of efciency. Therefore, to mitigate these losses, Soxhlet
extractor was used. The Soxhlet extraction is also a leaching pro-
cess which allows the indirect contact of the condensate of solvent
vapour with the solid (kept in a thimble) and the condensate of the
vapour was returned repeatedly to the solvent tank. The driving
force of mass transfer is maintained at the same level because the
fresh condensate of solvent is always used for extraction (De Castro
and Garcia-Ayuso, 1998). This was equipped with a reboiler at the
bottom and a condenser at the top to condense the vapour and drip
it down to the thimble. The thimble was stuffed with 29.2 g of soap
pulp. 200 ml of distilled water was taken in the still pot to start
the extraction process. Extraction temperature was xed at 100 C
(boiling point of water). During each cycle, a portion of the non-
volatile compound, i.e., soap pulp was dissolved in the solvent until
the solution in the distillation ask became concentrated. Finally,
the extract was allowed to cool and an E-24 test was carried out
with this concentrated solution.
2.4. Determination of emulsication index (E-24 index)
E24 index was determined following a standard protocol
(Cooper and Goldenberg, 1987). 2 ml of extract corresponding to
different extraction time and temperature was mixed with 2 ml
of parafn liquid in different test tubes. The tubes were placed in
922 A. Basu et al. / Industrial Crops and Products 77 (2015) 920931
Fig. 1. Main structures of Sapindus saponins extracted from the fruit pericarps of Sapindus mukorossi Gaertn.
a vortex (REMI CM 101Cyclo mixer) and spun for 2 min for vig-
orous mixing. The resulting mixture,which was allowed to stand
overnight, separated into an emulsied layer (hydrophobic phase)
and the remaining aqueous layer (hydrophilic phase). Finally emul-
sication index (E-24) was measured as the ratio of the height of
emulsied layer to the total height (Lawrance et al., 2014; Khopade
et al., 2012).
Height of emulsied layer
Emulsication Index(E 24) = 100
Total height of the solution
(1)
2.5. Determination of surface tension and critical micelle
concentration
The surface tension using soap nut extract, 2-HBP solution
and Ezee (commercial surfactant) solution were measured using
Leconde Du Nouy Tensiometer (Ring method) where the tension
required to lift the platinum ring out of the surface of the water
was measured (Sarma et al., 2012).
Fmax
Surface tension() =
(L cos())
where Fmax is the force exerted on the platinum ring, L is the char-
acteristic length of the ring and  is the contact angle between the
platinum ring and solution.
Generally,  = 0 for materials with optimum wetting properties
like platinum. At rst the ring was lowered so that it just touched
the surface of the solution taken in a beaker. Then the ring was
pulled upwards so that the liquid lm produced in the ring was
stretched. This tension created a force which was proportional to
the surface tension.
By plotting the surface tension curve using the values of surface
tension measured at different surfactant concentrations, Critical
Micelle Concentration (CMC) was determined with the onset of
constancy in surface tension inspite of change in concentration.
2.6. FTIR analysis
The aqueous soap nut extract was analyzed using an FT-IR
instrument (JEOL-JSMC 6700 F) and the FTIR spectra were gen-
erated under transmittance mode (4000500 cm1 ). A thin liquid
membrane of the soap nut extract was formed by dripping several
drops of the soap nut extract onto an aperture plate placed at the
bottom and sandwiching it under another plate placed on top. The
screws were tightened adequately, thereby forming a liquid cell.
Then infrared radiation was passed at the mentioned wavelength
through the liquid cell which absorbed radiation corresponding to
specic frequencies, thereby generating the required spectra (Shi-
madzu, liquid cell method). (http://www.shimadzu.com/an/ftir/
support/ftirtalk/talk9/intro.html)
(2)
3. Results and discussion
3.1. FTIR analysis (fourier transform infrared spectroscopy)
The FTIR spectra (Fig. 2) reveals the presence of alcoholic -OH
due to O H stretching which is conrmed by the presence of broad
peak at 3370 cm1 . Another peak is obtained at 1050 cm1 which
 A. Basu et al. / Industrial Crops and Products 77 (2015) 920931
Fig. 2. FTIR spectrum of soap nut extract.
is due to the C O stretch of carbinol. C O stretching at 1650 cm1
conrms the presence of carboxylic groups. However, some weak
peaks at 1450 cm1 are obtained which are due to C H bending of
alkanes. Intense peaks were obtained at 2100 cm1 due to the pres-
ence alkynes. The peaks are similar to those of saponin, as reported
in the literature (Sharma and Paliwal, 2013; Deore and Khadabadi,
2010). This is a qualitative indication of presence of saponin. The
present study is concentrated on the production of soapnut extract
to be used as a bio-emulsier-cum-surfactant. Therefore, the puri-
cation of saponin is out of the scope of this study.
Fig. 3. E-24 index of soap nut extract vs extraction time.
923
3.2. E-24 index
In Fig. 3, emulsication index of soap nut extract has been
plotted against extraction time using extraction temperature as
a parameter. The E-24 index of soap nut extract increases with
extraction temperature in all cases and nally becomes constant
while E-24 is found to be decreasing with time after a certain limit.
This may be due to the fact that with the increase of temperature,
the structure of soap nut is opened up and hence the extraction
becomes easier. On the other hand, at a particular temperature,
it is expected that extraction increases with increase of exposure
time. However, an overexposure to a particular extraction condi-
tion for a long time may become futile due to transfer of all saponins
to the extract phase. Overexposure may also be detrimental for
extraction due to denaturation of surface active component in the
heating mantle experiment. Fig. 3 clearly reects these phenomena
by showing the constancy in the E-24 versus time plot at tem-
peratures of 90 C and 100 C beyond 10 min (81.82%) and 7.5 min
(82.61%) respectively and decreasing trend of E-24 beyond 12 min
at 60 C, 70 C and 80 C respectively.
3.3. Statistical analysis of E-24 value and yield
In this study, the effects of time and temperature, which were
chosen as the independent variables, on the production of surfac-
tant from soap nut have been investigated and the emulsication
index and yield are optimized using Central Composite Design
(CCD) in Design Expert 8.0.7.1. The maximum and minimum values
are designated as +1 and 1 levels respectively. The axial points are
located at (, 0,0), (0, , 0) and (0,0, ) where is chosen to
be 1 which makes the design face-centered(Sugashini and Begum
2013). The +1 and -1 levels and the values of the two independent
variables are shown in Table 1.
The total no. of experiments to be carried out (Ferreira et al.,
2004) is calculated from the Eq. (3)
N = 2x + 2x + N0 (3)
924 A. Basu et al. / Industrial Crops and Products 77 (2015) 920931

Here N corresponds to the total no. of experiments, x is the no.
of independent variables and N0 is the no. of center points (N0 = 5).
A total of 13 runs was carried out which are presented in Table 2.
Generally the dependency of independent variables on the out-
put is given by
y = f (x1 , x2 , x3 ) + e
where y is the output, l is the power of the output, f is the function
of input variables x1 , x2 , x3 and e is the error. To nd the correct
transformation, Box-Cox plot (Fig. 4) has been used for the E-24
index model where the natural logarithm of the residual sum of
squares has been plotted against .  is chosen such that the value
of the residual is minimized, which is depicted by the green line
(Kobya et al., 2014). Since the  value does not fall in 95% condence
level, so the  is recommended to be 1 by the software.
The polynomial quality is determined by the determination
coefcients, R2 and Radj 2 which are given in Eq. (5) and (6) (zer
et al., 2009).
SSresidual
R2 = 1
SSmodel SSresidual
SSresidual /DFresidual
Radj 2 = 1
(SSmodel + SSresidual )/(DFmodel + DFresidual )
In these equations, DF is the degree of freedom and SS is
the sum of squares. The values of R2 and Radj 2 are presented in
Tables 3a and 3b for E-24 index and yield of soap nut extract
respectively. Since we aim at maximizing the values of R2 and
Radj 2 , it clearly indicates that quadratic equation is the best t
for both cases,since the corresponding R2 = 0.9769, 0.9734 and
Radj 2 = 0.9604, 0.9544 for E-24 index and yield model. Although R2
and Radj 2 of the quadratic model is highest among all the models in
case of yield, Rpred 2 (0.751) is much less than Radj 2 which may intro-
duce a large block effect in the model. As a result, a linear function
of time and temperature is considered to be the best representation
of yield.
The experimental data were tted using multiple regression to
the quadratic equation which is given by
2
Y = a0 + a1 X1 + a2 X2 + a12 X1 X2 + a11 X1 + a22 X2 + e 2
Table 1
Independent variables of the experiment and their corresponding levels.
Units 1 level +1 level alpha
Time Minutes 5 15 5 15
Temperature Centigrade 60 100 60
where X1 and X2 are the independent variables, Y is the output
variable, a0 , ai and aii are intercept, linear and quadratic con-
stant coefcients respectively, aij is regression 2-factor interaction
between X1 and X2 and e is the residual part (Stamenkovic et al.,
2013).
To analyze the effects and interactions, analysis of variance
(4) (ANOVA) was done for both of the models which are shown in
Tables 4a and 4b. In ANOVA, any factor with p-value less than 0.05
have signicant contribution to the response with 95% condence
level (Roosta et al., 2014). The E-24 index model F-value of 59.24
and p-value less than 0.0001 imply that the model is signicant.
Also the linear terms like A and B (A: time and B: temperature) and
the quadratic term A2 have signicant contribution to emulsi-
cation index. But the 2 factor interaction (AB) and the quadratic
term B2 are insignicant since their p-value is greater than 0.1.
This shows that extraction time has a greater signicance on the
model rather than extraction temperature, which justies the fact
that there may be some denaturation of soap nut after a certain
time limit. The F-value of the model increases to 77.93 and the
predicted correlation co-efcient (Rpred 2 ) increases from 0.7689
(5) to 0.8997 upon excluding the insignicant terms (Fermoso et al.,
2010).
Adeq Precision measures the signal to noise ratio, which is
(6)
desired to be greater than 4 (Abbasi et al., 2013). This model has a
ratio of 24.474 which indicates an adequate signal. The predicted
response values of the model are well in agreement with the actual
values which is shown in Fig. 5(a) and therefore determines a good
t of quadratic model. Fig. 5(b) depicts the residual vs predicted
response plot where the residuals are scattered randomly indicat-
ing that the variance is constant for all experiments (Idris et al.,
2006; Kamankesh et al., 2013).
The nal model equation correlating E-24 index for saponin
extraction as a function of time and temperature, is obtained as
follows
EmulsicationIndex = 78.71+9.15A + 9.61B + 1.44AB 13.11A2
2.42B2 (8)
where A and B corresponds to time (min) and temperature ( C)
(7) respectively.
In yield, model F-value of 17.62 and p-value equal to 0.0005
imply that the model is signicant. Also time and temperature sig-
nicantly affect the yield of extract as their p-value are less than
0.05. Adequate precision in this case is achieved to be 14.15. The
+alpha predicted vs actual plot (Fig. 5c) and residual vs predicted response
plot (Fig. 5d) for yield conrms the linear t to be quite satisfac-
100 tory. The yield of soap nut extract can be represented as a function
of time and temperature as follows

Table 2 Yield = 0.21 + 1.002A + 0.19B (9)

RSM design and the experimental valuesof yield and emulsication index of
extracted soap nut solution.
3.4. Optimization of emulsication index and yield of soap nut
Run Time (mins) Temperature Emulsication Yield of soap nut extract
( C) index (%) extract (gm extract/gm
raw pulp)
For the optimization of emulsication index and yield simulta-
1 5 80 55.00 19.14 neously, 3-D surface plot has been done with time and temperature
2 10 80 78.95 27.48
as the parameters. The E-24 index plot (Fig. 6a) gives a concave
3 10 100 81.82 30.09
4 10 80 78.95 27.48 plot, suggesting that at a particular time and temperature, the E-
5 10 80 78.98 27.48 24 value attains a maximum and then decreases slightly. In case of
6 5 100 64.65 23.78 yield, the plot (Fig. 6b) shows that the yield of the extract increases
7 5 60 44.83 14.72 with increase in time and temperature.Contour plots (Fig. 6c and
8 15 100 85 34.26
9 10 80 78.95 27.48
d) depict the lines where E-24 and yield are constant throughout
10 10 80 78 27.48 with respect to different time and temperature.
11 15 80 75 26.10 Now the optimum extraction and the E-24 index are achieved
12 10 60 69.56 22.84 at different values of time and temperature. Thus a compro-
13 15 60 59.40 20.50
mise between the values is achieved by the desirability function
 A. Basu et al. / Industrial Crops and Products 77 (2015) 920931 925

Fig. 4. Box-cox plot for power transforms for selecting  of the E-24 index model.

Table 3a
Model summary statistics to predict the best t of the E-24 index model.

Source Standard deviation R-squared Adjusted R-squared Predicted R-squared Comments

Linear 8.38 0.6010 0.5212 0.2117 Suggested

2FI 8.78 0.6057 0.4743 0.6739
Quadratic 2.41 0.9769 0.9604 0.7689

Table 3b
Model summary statistics to predict the best t of the yield model.

Source Standard deviation R-squared Adjusted R-squared Predicted R-squared Comments

Linear 2.61 0.7789 0.7347 0.5730 Suggested

2FI 2.64 0.7968 0.7291 0.2918
Quadratic 1.08 0.9734 0.9544 0.7510

Table 4a
Analysis of variance (ANOVA) for predicting the model of E-24 index vs time and temperature.

Source Sum of squares df Mean square F value p-Value Prob > F Comments

Model 1718.51 5 343.70 59.24 <0.0001 Signicant

A-time 502.70 1 502.70 86.64 <0.0001 Signicant
B-temperature 554.50 1 554.50 95.57 <0.0001 Signicant
AB 8.35 1 8.35 1.44 0.2693
A2 475.04 1 475.04 81.87 <0.0001 Signicant
B2 16.24 1 16.24 2.80 0.1382
Residual 40.62 7 5.80
Lack of t 40.62 3 13.54
Pure error 0.000 4 0.000

Table 4b
Analysis of variance (ANOVA) for predicting the model of yield vs time and temperature.

Source Sum of squares df Mean square F value p-Value Prob > F Comments

Model 240.69 2 120.34 17.62 0.0005 Signicant

A-time 150.73 1 150.73 22.07 0.0008 Signicant
B-temperature 89.96 1 89.96 13.17 0.0046 Signicant
Residual 68.31 10 6.83
Lack of t 68.31 6 11.38
Pure error 0 4 0
926 A. Basu et al. / Industrial Crops and Products 77 (2015) 920931
Fig. 5. Plot of predicted vs actual response (a), residuals vs predicted response (b) for E-24 index, predicted vs actual response (c), residuals vs predicted response (d) for
yield.
(Derringer 1980; Zhu et al., 2014). The responses are rst con-
verted to a desirability function (dk ) which scales from 0 to 1. If
the response is outside the acceptable region, dk is equal to 0 while
any fully desirable response sets dk as 1.
The individual desirability (dk ) is dened by:

0
wk whenyk ykmin
yk ykmin
dk = whenykmin yk ykmax
ykmax ykmin
1 whenyk ykmax
where yk is the value of the response, ykmin is the minimum accept-
able value of yk and ykmax is the maximum acceptable value, wk is
the weight assigned to the response. Here ykmin and ykmax are taken
as the minimum and maximum experimental values of the E-24
index respectively.
The overall desirability (D) is taken as the geometric mean of the
individual desirabilities which is given by
v1 v2 v 1/n
D = (d1 d2 ....... dn n )
where vk represents the importance of desirability dk and 0vi 1.
n indicates the total no. of individual desirabilities. Thus using the
desirability function, maximum E-24 value that can be attained, is
found out to be 88.042% at 100 C and 12.02 min with a desirability
of 0.783 as indicated in Fig. 7. On the other hand, the optimum con-
dition for yield (35%) has been observed to be at 15 min and 100 C
when E-24 index decreases to 83%. From the experimental study
and the optimization study it is clear that there is no betterment
of the value of E-24 with the increase of time beyond 12 min. The
saponin may be affected by the over-exposure to high temperature
environment for a long time due to heat sensitivity. To avoid the
detrimental effects of overheating and long processing time on the
properties of target biomolecule, i.e., saponin, the optimum condi-
tions for E-24 has been used as the most desired process condition
where the yield is 31.21%. This condition has also been used during
the cost analysis.
(10)
3.5. Soxhlet extraction of saponin from soap nut
The Soxhlet extractor produces a soap solution that approxi-
mates the maximum attainable concentration of saponin in water
under optimum conditions. Also, the apparatus is so designed as
to prevent losses due to vapour leakage. A mass loss of around
3.2% of the initial mass is however, encountered. The E-24 index
using this setup, is observed to be 87.23%, which is almost equal to
the optimized E-24 value of extract via evaporative extraction. The
(11) experiment thus proves that a reasonably high value of E-24 indi-
cating a high surfactant activity, can be attained within a reasonable
time span and extraction temperature.
3.6. Surface tension and CMC
The surface tension has been observed to decrease with increase
in concentration of surfactant in all cases (Fig. 8) until the criti-
cal micelle concentration is reached. The CMC values have been
 A. Basu et al. / Industrial Crops and Products 77 (2015) 920931 927

Fig. 6. 3-D Surface Plot (a) of E-24 index and (b) of yield vs extraction time and temperature, contour plot (c) of E-24 index and (d) of yield vs extraction time and temperature
for soap nut solution.

Fig. 7. Optimum extraction of soap nut as a function of time and temperature.

928 A. Basu et al. / Industrial Crops and Products 77 (2015) 920931

Fig. 9. Comparative study of E-24 index of commercial detergent (Ezee), 2-HBP

solution and soap nut extract.

obtained as 23 kg/m3 (2.3%) for optimally extracted saponin rich

soap nut solution (Fig. 8a), 0.2 kg/m3 for 2-HBP (Fig. 8b) and
250 kg/m3 (25%) for Ezee (Fig. 8c). The corresponding surface ten-
sion values are measured to be 40 mN/m, 30 mN/m and 18 mN/m
for soap nut, 2-HBP and Ezee respectively at their critical micelle
concentration.
Ghagi et al. (2011) and Sarma et al. (2012) reported the CMC
value of 17 kg/m3 for soapnut extract corresponding to a surface
tension of 38 mN/m. Although the surface tension obtained at CMC
is comparable, the value of CMC in the present case is higher than
the reported one. In those research studies, no extraction proce-
dure is followed. Since the present work is based on the extract
of soap nut and not raw one, as reported in both previous works,
there is a difference in the CMC value. In another research article
Balakrishnan et al. (2006) reported the CMC value of 0.45% (w/w)
for pure sapindus saponin using conductivity meter. These differ-
ences may be due to the fact that the extract in the present study is
not constituted purely of saponin. The surface tension may vary due
to interference of different chemical compounds which are present
in the extract with saponin.

3.7. Comparison of E-24 value with conventional surfactants

An E-24 index test was carried out for the aqueous solution of
commercial surfactant (Ezee) and 2-HBP which is expected to enter
the surfactant market in the form of detergent as competitors of
conventional detergents and the bio-detergent like reetha extract
etc. with the popularization of biodesulfurization of diesel produc-
ing 2-HBP as a byproduct (Bandyopadhyay et al., 2013). The values
have been determined to be 79.6% and 64% respectively. Therefore
the E-24 index of optimally extracted soap nut solution is higher
than both the commercial and waste-derived surfactant under the
study (Fig. 9). It may therefore be expected that there is a prospect
for commercialization of extract of soap nut, derived using simple
device, for substitution of the commercial detergent.
Fig. 8. Surface tension plot of (a) Soap nut extract, (b) Pure 2-HBP in acetone solu-
tion, (c) Commercial Detergent (Ezee) solution.
3.8. Cost analysis

As the commercialization of any product depends on its produc-

tion cost to attain a competitive position with respect to similar
products in the market, therefore, a cost analysis of production of
saponin rich extract from reetha has been done based on the data
 A. Basu et al. / Industrial Crops and Products 77 (2015) 920931 929

Fig. 10. Comparative study of annual cost of production of three surfactants extracted from Ezee, soap nut and 2-HBP.

of laboratory experiments. The data on cost, energy and operating
schedule of small scale plant are provided in Table 5.
From the cost calculations, it is determined that 32.5 kg of
saponin rich extract is produced annually with an expenditure of
Rs. 31,700 approximately. The detailed calculation has been pro-
vided in the supplementary section. The costs of same quantities
of 2-HBP and Ezee in the market are Rs. 107,200 and Rs. 6500
respectively. Fig. 10 also depicts the comparative cost analysis of
soap nut extract with 2-HBP and Ezee. From the cost analysis, it
is clear that although saponin rich reetha extract is cheaper than
2-HBP, it is costlier than the liquid detergent, Ezee. However, from
the perspective of E-24 value, soap nut extract leads the list of all
surfactants. The CMC value (23 kg/m3 ) of the extract is much less
than that of Ezee (250 kg/m3 ) and is relatively larger than that

Fig. 11. Annual cost of production vs CMC plot for all surfactants.
930 A. Basu et al. / Industrial Crops and Products 77 (2015) 920931
Table 5
Cost analysis of extraction of surfactant from soap nut in small scale.
Fixed cost
Cost price of each heating mantle
Recurring cost
Grid electricity tariff (Calcutta electricity supply
corporation)( /kWh)
Energy data
No. of heaters in operation
No. of heaters in stand-by mode
Average rating of heating mantle
Schedule of plant operation
Working hours per day
Working days per year
Miscellaneous cost (cost of glassware etc)
of 2-HBP. Coincidentally, the CMC value of the surfactants varies
inversely with their cost under the present study (Fig. 11) which is
represented by the Eq. (11).
 0.1125
Annual cost = 152, 300 CMC 75, 380
where annual cost is expressed in Rupees and CMC is expressed
in kg/m3 . Hence for the reduction of surface tension in the same
extent, the more expensive surfactant has to be utilized in much
lesser quantity. In this way, the cost and quality of the surfactants
may be balanced in detergent industry and for enhancement
of oil recovery. Moreover, as 2-HBP is derived from petroleum
based waste products, therefore there may be certain chemical
constituents which may restrict its application in certain specic
areas like food, pharmaceuticals etc. However, the leading position
of optimized reetha extract with respect to E-24 value and its
natural origin should make it an attractive product to be used for
emulsication in food and pharmaceutical industries.
4. Conclusion
The evaporative extraction of saponin has been optimized for
the rst time using statistical tool, namely RSM. Optimum values
of extraction temperature and time have been determined to be
100 C and 12.02 min respectively corresponding to an E-24 value
of 88.042%. The functionality of E-24 simultaneously on extrac-
tion time and temperature has been correlated mathematically for
the rst time where the extraction time has a signicant contribu-
tion in the model equation rather than temperature. The extract
obtained using the Soxhlet apparatus also exhibited the same E-24
value as the optimally extracted solution using thermal arrange-
ment indicating the similar concentration of saponin in it. The CMC
value (23 kg/m3 ) of saponin extract obtained using heating mantle
arrangement is comparable to those observed by other investi-
gators. When compared to the standard surfactants, it has been
observed that although the CMC value of the optimal extract is
higher than 2-HBP (0.2 kg/m3 ), it is much lower than the commer-
cial detergent, Ezee (250 kg/m3 ). The E-24 value of saponin extract
is highest among those of all the three surfactants under study.
A cost analysis of extraction of natural surfactant from reetha has
been performed. The production cost of reetha extract is higher
than Ezee but much lower than 2-HBP. The cost of a surfactant
inversely varies with its CMC value following a correlation. Thus
the requirement of reetha extract is much less than that of Ezee for
the reduction of surface tension upto the same extent and much
more than that of 2-HBP. It is expected that using the research
outcomes of the present study, saponin rich extract may be eco-
nomically produced from abundantly available Indian reetha seeds
using either optimum conditions of evaporative extraction or in a
Soxhlet extractor in commercial scale and it will serve prominently
as a surfactant in detergent industry as a unique emulsier well
as in food and pharmaceutical industries where the use of petro-
derived emulsier is impossible due to health issues. However, the
extraction processes studied under the present study will be par-
1900 ticularly attractive for small scale industries because of their simple
methodology requiring very little training (De Castro and Garcia-
6.77 Ayuso, 1998). For further sophistication in large scale industries,
processes using ultrasound, microwave (Wu et al., 2015) and super-
critical uid extraction may be utilized after rigorous energy, cost
33100 W
and environmental analyses.
10 270 Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
Rs. 5000
the online version, at http://dx.doi.org/10.1016/j.indcrop.2015.10.
006.
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