J.

of Supercritical Fluids 67 (2012) 123130

Contents lists available at SciVerse ScienceDirect

The Journal of Supercritical Fluids

j o u r n a l h o m e p a g e : w w w. e l s e v i e r.c o m / l o c a t e / s u p f l u

Optimization of supercritical carbon dioxide extraction of essential oil from

spearmint (Mentha spicata L.) leaves by using Taguchi methodology
Karamat Ansari a, , Iraj Goodarznia b
a
b

Chemical Engineering Gachsaran Gas and Oil Production Company (GS NISOC), Gachsaran, Iran
Chemical & Petroleum Engineering Department, Sharif University of Technology, Tehran, Iran

a r t i c l e

i n f o

Article history:
Received 15 February 2012
Received in revised form 18 March 2012
Accepted 19 March 2012
Keywords:

Supercritical extraction
Spearmint
Total yield
Essential oil and Taguchi method

a b s t r a c t
In this study, the essential oil of aerial parts species of a plant called Spearmint was extracted by CO2
to optimize the results of the supercritical extraction process. In order to achieve maximum total
yield extraction and SF-CO2 concentration, tests were done in a laboratorial pilot considering the
Taguchi method under following condition: pressure. 90,100,140,170 bar, temperature. of 35, 40, 45, 50
C, mean particles size: 250,500,710,1000 m, ow rate 1,3,5,8 ml/s and dynamic time 30,50,90,120 min.
The optimizing conditions for SCE and SF-CO2 concentration are: 90 bar, 45 C, 500 m, 5 ml/s, 120 min and 90
bar,
35 C, 250 m, 1 ml/s, 30 min. SCE total yield and
concentration are 2.032% and 0.033 mg/ml. In simCO2
ilar and optimal conditions of extraction, supplementary tests were done so resulted and experimental
amount of the calculated sum are in accordance with total yield extraction and SF-CO2 concentration.
This is the rst report announcing optimization of spearmint SCE by use of Taguchi method.
2012 Elsevier B.V. All rights reserved.

1. Introduction
Lamiaceae family is one of the biggest herbal families that
have geographical and universal dispersion. This type is grown
in all around the world apart from Antarctic and northern region.
These families consist of about 200 genuses and between two or
5000 species of fragrant plant and short shrubs having a large
diversity [13]. Respectively, morphological characteristics of Lam
families includes quadrangle stems, simple leaves arranged in
vertical direc- tion that have the reciprocal and alternative
forms, with ve-part sepal and two or four stamens ocymun
bacilicum, rosmarinum ofcinalis, Thymus vulgaris, Origanum
vulgare and Menthe belong to the same family [4].
The menthe or mentha genus, that has a vast diversity and
importance, is one of the most important one in this family used
by human being over 2000 years that is grown in all of 5 continents having remarkable and commercial importance. Menthe
has thirteen hybrid named and composed of 19 geographical
diversity [57]. This menthe genus has 2530 particular species
growing in Europe, Asia, and South Africa [8]. Some of the
important species of this genus are Mentha spicata L., piperita L.,
aquatica L., pulegium L., etc. that grow in all around the world. The
M. spicata L. or spearmint species or with the abbreviated name
mint is the same green men- the or edible menthe that is used
more frequently. Spearmint is

Corresponding author. Tel.: +98 7429122461; fax: +98 7422224015.

E-mail address: ansari karamat2006@yahoo.com (K. Ansari).

0896-8446/$ see front matter 2012 Elsevier B.V. All rights reserved.

a cross breeding species resulted from M. longifolia and rotundifolia. Menthe is one of the herbs that are so important because of its
essence and pharmaceutical compounds.
Because of its benets and economical values the menthe is one
of the most important herb and many researchers have focused
their attention on it in the way they can increase its yield and
they can know the important factors involving the quality and the
quan- tity of the extraction essence. The green menthe or the
edible one that is the other species of this family, has more usage
[7,913].This family and this herbal species of mint is the rich
source of poly phenolic compounds and, therefore have a strong
antioxidant prop- erties used in alimentation industry that
prevent the oxidation. This material can be a suitable substitute
for articial and industrial antioxidants like butylated hydroxyl
toluene (BHT), and butylated hydroxyl anisole(BHA). Because of
the harmful effect of articial and industrial antioxidants on
human beings health; the natural alternative is more
emphasized [1416].
The recent studies show that the Lamiaceae is a potential
resource for natural antioxidants. This property depends on some
compounds such as avonoids, diterpenes, phenolic and phenolic
acid [1722]. The antimicrobial property of both species of menthe
herbal essence has been proven and has a vast industrial, pharmaceutical, and cosmetic use and its consumption is increasing in
the world [2327]. The menthe is largely used in the conventional
medicine because of its antispasmodic, antiseptic, and emmenagogue effects. The spearmint essential oil is used for the different
kind of chewing gums, beverages, cosmetic products, perfumery,
toothpastes and mouth washes [28,29].

doi:10.1016/j.supu.2012.03.011

124

K. Ansari, I. Goodarznia / J. of Supercritical Fluids 67 (2012) 123130

This valuable essence has important compounds such as carvone, Methone and d-limonene; itself is a strong insecticide and
that has mutagenic activity. Also this essence has anti-fungal,
antiviral, treat fever, diuretic, stimulant, and bronchitis properties,
which are suitable for cramp, gastritis, headaches,
indigestion, nausea and lots of other cases [22,23,3033].
So far different kinds of techniques have been used for the
extraction of the herbal essence and the isolation of the
important essence. Distillation manners, steam distillation,
hydro distilla- tion, and organic solvent extraction that consist
of varied set of maceration percolations and soxhlet techniques
used since long time conventionally. Conventional Sechelt
extraction is a stan- dard common technique that has a lot of
problems such as long period of extraction, high consumption of
chemical solvent, prob- lem of solvent separation, toxic solvent
and high cost of energy. The manners of distillation used in
high temperature, are not suitable for the valuable compounds
that are sensitive to temperature and can be resulted in reduction or loss of product.
The supercritical extraction methods that is completely new than
these methods has not the problems of earlier and conventional
techniques. Because of its unique properties such as high solubility, low viscosity of supercritical uid and ne selectivity
property that present a vast use
in alimentation and
pharmaceuti- cal industries and the extraction of natural and
herbal raw materials [3439].
CO2 with critical condition C(0.47 mg/ml), T(31.18 C),
P(73.8 bar) has many interesting properties such as safety,
inammability, and inexpensiveness which like the other supercritical uids has no free residue in product having
environmentally friendly character that is highly suitable for
the supercritical extraction. Despite of multiple problems and
because of the simple and facility of use, the manner of
extraction with sol- vent is common yet [40,41]. So far the
extraction of the herbal essence of M. spicata L. has been
reported
in
conventional common manner
such as
distillation and the
extraction with different
chemical
solvents and in some cases the supercrit- ical extraction of
herbal essence has been included. In some cases of the
supercritical extraction, the co-solvent or mod- iers (cosolvent) has been used for increasing the yield of extraction
[17,23,25,27,36,4245].
In some published articles and most review papers related
to supercritical extraction of the different herbal species in
which, the extraction of herbal essence of M. spicata L. has
been dis- cussed using mathematical modeling, the extraction of
edible an herbal menthe has been also studied [19,24,45]. In
some of pub- lished articles related to the supercritical
extraction of this herbal species of spearmint, the design of
experiment has been done and we have calculated the maximum
yield of the herbal essence and the optimal condition of the
extraction. In spite of the fact that researches done in this area,
it seems optimization of supercriti- cal extraction of spearmint
essential oil has been performed using Taguchi testing method
was not done yet
and this is the rst report announcing
[17,25,27,36]. Fig. 1 shows this valuable herbal plant.

2. Materials and method

Herbal samples of M. spicata L. were cut and collected as the
aerial part and near to the period of owering at the end of
summer from the farms of south west of Iran in Kohgiluye and
Buyerahmad province. The samples were very fresh and fragrant
and dried in shadow along with dry weather with low moisture
and have been kept in suitable container and lace before the
beginning of the tests. The time of drying lasted about 2 weeks.
The wet samples had more than 80 percent humidity.

Fig. 1. Plant genus of M. spicata L (spearmint).

Table 1
Average diameter particles size and weight samples.
Sample no.

Mean particle size ( m)

Weight (g)

1
2
3
4

250
500
710
1000

164.15
263.33
105.42
111.08

643.98

Sum

2.1. Sample preparation

We have grinded the dried herbal samples of spearmint near
to the period of tests performance in one suitable mill carefully
and with patience in a way that the temperature of the samples
in order to preventing the increase of temperature. Each sort of
temperatures increase of the samples results reduce of quantity
and quality of extractive essential oil. We have prepared the
powder using suitable sieves and shaking machines in four sizes
and the granulated samples were stored in 4 glass container. In
Table 1, the mean particles size of samples and the amount of
their weight have obtained. To regulate the time of grinding, it can
achieve the approximated equality of weight in samples that is
so important for all the tests.
2.2. Material and reagent
Some of the consumable materials are necessary for all the
supercritical tests. Carbon dioxides storages with 99.9% of purity
have been prepared in Rohams factory. Sealed dark bottles with
caps in little volume (1 ml) for preserving products essence, special tissues for the sacs in which the powder of tests have been
reserved, chemical materials and required solvents such as ethanol
and acetone all have been purchased from the market.
2.3. Design of experiments
Supercritical extraction depends on different factors and
despite of its multiple advantages, also because of the difculties
and test performance complexities in high pressure and in order
to reduce the number of tests, it is necessary to design of the
experiment (DOE). In general, DOE can reduce the costs and
economize the time. Also, DOE can be done in different manners.
Generally In the process of the supercritical extraction of the
herbal essence, the most important factors concerning the total
yield of extraction and the concentration of the essence in the
supercritical uid are: pres- sure and the temperature of the
extraction, mean particles size of feed powder, ow rate or the
velocity of supercritical uid and the time of the test or dynamic
time.

Table 2
Variable parameter range in SCE for spearmint essential oil.
No.

Parameters

Unit

Variable range

1
2
3
4
5

Pressure (P)
Temperature (T)
Mean particle size (Di.)
Flow rate (QCO2 )
Dynamic time (D.t.)

bar

C
m
ml/s
min

90 < P < 300

35 < T < 60
250 < Di. < 5000
1 < QCO2 <
30 < D.t. < 120

Table 3
Experimental levels of the factors used in Taguchi method for SCE of spearmint
essential oil.
Factors (parameters)

Pressure (bar)
Temperature ( C)
Mean particle Size ( m)
Flow rate (ml/s)
Dynamic time (min)

Factors levels
1

90
35
250
1

100
40
500
3

140
45
710
5

4
170
50
1000
8

30

50

90

120

Almost in the all of former researches on the supercritical

extrac- tion and such as researches done on the spearmint
essential oil supercritical extraction, we have used the same ve
parameters or some of them. Of course for this special purpose,
most of stud- ies have been done based on three parametric tests.
According to the researches on this herbal species with the
characteristics of the our laboratorial pilot and estimate/error test
in different condi- tions, we have determined the domain of
parameters changes, that summarized in Table 2 [17,25,27,36,43].
In this experimental research, in order to design of the experiment, we have used Taguchi method because of facility and
simplicity of use. This method is so precise and reliable, having
different use in science and engineering. When the number of
ef- cient parameters is more than three, the use of Taguchi
method is suitable for designing the tests because of economy in
time, cost and depreciation of laboratory devices. Furthermore,
the result of this method, highly depend on experience and
further test, ensur- ing better result. Also, if necessary, it can
repeat the test in different array and conditions in order to get
better results [4648].
According to this research, in Table 3, the efcient factors on
quantity and quality of extractive essence in supercritical manner and total yield of extraction process consist of: pressure and
temperature of the extractions utensil, mean particles size of feed
powder, ow rate of supercritical period, and dynamic time, each
one is seen in four different levels. L16 array has been prepared
based on this table.
2.4. Total yield extraction and supercritical uid concentration
Total yield of supercritical extraction has been dened in different manners. In researches works that most insist more on
extractions modeling and equilibrium state, the yield is calculated
by dividing the amount mass of extractive essence in supercritical
process on maximum mass of extractable essence or the amount
of primary mass in feed powder. Any report in which
experimental aspects of
process and the optimization of
supercritical extrac- tion are discussed, denes total yield with
dividing the mass of extractive essence on the mass of consumed
feed. In both of these manners, this ratio is expressed as
fractional or percent number. In this study we have used second
manner and the yield is dened as the gram amount of produced
essence on the gram amount of consumed feed powder.
The concentration of supercritical uid that shows the amount
of dissolved herbal essence in the supercritical uid in different
laboratory conditions has always the same criteria and can just
have

different units. In this research, the concentration of essential oil

in supercritical uid CO2 has been expressed as g/L or mg/ml. To
achieve the maximum yield of extraction process and maximum
density of supercritical uid are the most important goal. The total
yield resulted number that is in fraction and percent is very little
and can be dened as mg/g (milligrams of extractive essence
divide on gram consumed food) [17,25,4951].

2.5. SCE process optimization and statically analysis

Generally, in optimization, it came to be considered the
achieve- ment of goal with nearest, best and cheapest route or
maximum
and
minimum of one special product in the process. For the
harmful minimum range of production or its deletion was the
products,
rst goal. In order to produce valuable and expensive materials,
maximum range of production with low cost has been the rst regard.
In clinical researches such as supercritical extraction in which the
costs of tests are high, regarding the high pressure that is out of
criti- cal domain and about which the safety considerations are
exclusive and some of the special tests whose performance
demand much attention, it is necessary to apply all of the safety
considerations in order to optimize the process of this tests that
may be in some cases the only choice and issue. The
optimization can be done in different manners like the design of
experiment. In some cases such as complete randomized design
(CRD) and response surface methodology (RSM), when we have
the tests with multiple param- eters, the calculations will be
more complex and time-consuming [17,25,36,52,53].
The use of Taguchi method in order to design and optimize the
supercritical tests besides simplicity has reliable, good and exact
results. According to the parameters and the levels of change (5
factors, each one in 4 level) in this manner we have as below:
In order to carry out the tests, we have used orthogonal array
(L16).Every time, we change laboratory conditions, considering
six- teen times of test we can calculate total yield of extraction
process and supercritical uids concentration. To avoid different
errors, it can repeat each run two or several times.

(1) An optimum condition of supercritical extraction is determined

for achieving maximum yield of extraction and supercritical
uids concentration.
(2)
Optimum response or nal yield of extraction and
supercritical uids concentration are
estimated and
calculated.
(3) To ensure the calculated and assessed results with the same
optimal conditions in item no. 2, we have done conrmation
experiment.

The determination of optimum conditions and the optimization of supercritical extraction and the calculation of maximum
response in total yields extraction and supercritical uid concentration have been done with use of qualitek-4 software and the
analysis of variance or ANOVA (items no. 2 and 3). Also, it can
determine the effect of different parameters on optimum response
of extraction. The use of this software and ANOVA provides the
conditions in which it is possible to reexamine or conrmation
experiment and become aware of the accuracy of optimum
amount.
In the case of contradiction with estimated and calculated
amounts and in order to reduce or delete the errors, it can change
the conditions of the test, in other words it is possible to change
the levels of parameters or revise the test and get to better
results. On the other hand, this test presents (Taguchi method)
different advantages that help to identify and delete the most
efcient ones; it is an important subject [54,55].

2.6. Static
SCE

and

dynamic times in

It can dene static time as a time in which the conditions of

the test has been regulated and there were no uid ow rate in
the system. In static time, solute component or components
penetrate into the supercritical uid. This solubility can be
equilibrated and then does not increase, if selected time were
correct. In this time we have no production of essence. The
extraction of essence has done in dynamic time; itself is the time
of supercritical extraction test. The effect of suitable static time in
the period of static has two economical results:
(1) The increase of total yield of supercritical herbal essence
extrac- tion.
(2) The reduction of supercritical uid consumed CO2 volume.
The increase of static time more than required amount has no
effect on nal yield extraction. In previous researches that have
reported the extraction of herbal essence, it can select the static
time until 30 min, but for more condence and that the tests
achieve the static time or even pass it, for all the tests, based on
static time and temporal conditions, the time can be considered
until 50 min or an hour [17,25,36].
2.7. The SCE apparatus
For all the tests in this research, we have used a pilot whose
details is found in the references [56,57].This pilot has a multipurpose system with diverse functions of SCE. The extractions
container is cylindrical whose approximate volume is 35 ml in
which it can use 15 g of alimentary powder. The ability of pressure tolerance arrives up to 400 bar and through a good control;
operation of the supercritical extraction can be performed in temperature of 0100 C. The pump of pilot that is reciprocating, and
is
Haskel (USA Company) type. The other parts of pilot consist of
shell and tube extractor, CO2 uid dryer, tubing, etc. whose
materials are stainless steel 316.
This system works
discontinuously and extrac- tive essence is gathered in Pyrex
glass receiver. However, this pilot has many functions in different
elds of supercritical extraction experiments.
We have supplied CO2 gas storage in resources, with maximum
pressure of 80 Bar and the purity of 99/9% from Tehran Roham
Gas factory. The measurement of velocity and the consumed
volume of supercritical uid in this pilot have been done using
simple and precise instruments such as: wet test meter and
bubble ow meter
with high precision (2.831 105 m3 and 1 ml/s). In Fig. 2 we have
schematic shape of supercritical extractive pilot.
2.8. Experimental procedure
In order to do supercritical tests and considering high range of
pressure, the question of security in laboratory is very important
and because of low production of extractive essence, it should pay
attention to all steps of test avoiding and reducing eventual
errors. To extract herbal essence of edible menthe spearmint,
explained in Section 2.7, the instruction and the manner of test
are as below. At rst all the required materials should have been
prepared.
2.8.1. Cleaning and pilot equipment pretest
Before doing any test, especially the rst one, it should clean
the extractor and receiver with water, washing liquid and
chemical solvents such as acetone and alcohol. The CO2 gas
bomb should be full and have sufcient amount of gas pressure.
All of pilots Tools such as high pressure pump, cold and hot

uids circulation pumps, temperature

electric heater of water

control system (TIC),

resource, etc. should be turned on and off in order to ensure its

correct function.
2.8.2. Set up providing, leakage test and loading feed powder
After cleaning and testing the primary tools of system, it is necessary to test it for preventing CO2 leakage. To remove eventual
leakages, it should increase step by step interior pressure of system of several psi to several bars and it should consider manner
perfect works. According to L16 array, the pilot is prepared for the
rst run in 90 bar, 35 C, the mean particles size of feed powder in
sample no.1 (250 m), the ow of supercritical uid in 1 ml/s and
dynamic time of 30 min. Rotatory circulation pump of extractor
shell is turned on and the temperature control system is regulated
on 35 C. Now, we take apart feed extractor and we should ll it
with feed powders. The amount of consumed feed for the rst run
was 12.43 g and for the weight of feed and product, we have used
a digital balance with high precision. All the pressure and
temper- ature indicators, measurement system have been
regulated and calibrated and check tests before experiments
performance.
2.8.3. Internal purging system
By opening and closing each part of pilot, the air penetrates
into the system. It should evacuate this volume of air in each run.
The air with CO2 reduces the total yield of supercritical extraction
and itself causes error in the test. With low pressure gas of CO2
that is applied several times, the air is evacuated of the end of
pilot route. The operation of air evacuation in the system for the
rst run is more important and need more time rather than the
next tests.
2.8.4. Pressurizing and static time
After evacuating systems interior air and when the temperature of extractor arrives at 35 C, the needle valve is closed on
the exterior of extractions extractor, this operation increase gradually the pressure of test, with high pressure pump it can arrive
at favorite pressure. Here, we have static time and with adjusting
the pressure on 90 bar, the static time stares practically. Static time
is held at 50 min or an hour and in this time period, if the pressure decreases, it can retrieve this problem with a high pressure
pump that controls the temperature on 35 C. In this step of static
time, equilibrium is done in the extractions utensil and The S.t.
herbal essences molecules with supercritical uid of CO2 arrive at
saturation, then the mass transfer is stopped.
2.8.5. Essential oil production and dynamic time
At the end of static phase, dynamic time begins with opening
of exterior needle valve of extractor. In this step, we have essence
extraction. This step is the most difcult one in supercritical
extrac- tion test where pressure, extractions temperature and
ow rate should be controlled and preserved in favorite amounts.
The uid ow rate in run (1) is 1 ml/s. It should keep warm the
route with special mechanism in
order to
prevent the
obstruction of needle valve and formation of dry ice phenomenon.
The solution of super- critical uid with herbal essence of
spearmint penetrated enters the receiver after the needle valve.
In this extraction container, herbal essence is collected and CO2
gas pass through a pressure indicator, wet test meter and a bubble
ow meter and is vented. The end of dynamic time closes the
former and next valves of extractor, the conned pressure is
evacuated slowly. We have the same ve operations for the next
tests. The gained yellow essence is collected carefully, since it is a
little and very sensitive, after weighting is stored in the special
container of
which is a sealed dark bottle. Before analyzing, the bottles are kept
in a cold place without sunlight like fridge. At the end of rst run
and after getting the amount of produced essence and consumed
CO2 , it can calculate total yield of extraction and supercritical
uids concentration.

Fig. 2. Pilot plant of supercritical uids (CO2 ) extraction.

3. Results and discussion

Based on experiments designing in the framework of Taguchi
method and according to array and orthogonal L16, and
considering the change of condition and different parameters
such as pressure and time of extraction, supercritical ow rate,
mean particles size of feed powder and dynamic time, sixteen
times of test and just one repetition, we have done in sum 32
main tests. We have sub- sequent results in this section. The
study of different parameters, similar pressure, temperature,
dynamic time, etc. on supercritical extraction essential oil of M.
spicata L. (Spearmint) can be reported by another paper that will
publish.
3.1. SCE total yield results
At the end of each run, the total yield of supercritical extraction is resulted by dividing the amount of produced essence mass
on the mass of consumed feed powder of spearmint herbal
species that is expressed as a percentile, summarized in Table 4.
As we can see in this table, in the case of change in conditions, the
amount of total yield extraction is different and is varied of 0.25
1.82%. The run number (3) has the highest yield and in run 16 or
the last run, we have the lowest total yield of extraction. In both
of the runs, just the parameter of supercritical ow rate is xing
and the other factors of test conditions are different. Among the
different factors or conditions of these two runs with the
minimum or maximum nal yield, the pressure has the most
difference and the heat tem- perature and dynamic time each one
has one level of difference and the factor of feed powder has two
level of difference. From Table 4, one can observe that for each
test, the static mode time is xed on
5060 min or 1 h. Addition of a static step in to the extraction
con- dition did not have any effect on the recovery of essence oil
from plant genus of M. spicata L.
On the other hand, it can say that the parameter of pressure
has the most negative effect in the yield of supercritical
extraction, in

other words if we increase the pressure, the yield decreases,

super- critical ow rate has no effect as well, after that the factor
of mean particles size of powder has positive effect resulting the
increase of yield, in the case of increased feed powder diameter.
Both last parameters; temperature and dynamic time have an
equal effect on supercritical extractions total yield while heat
temperature has negative and dynamic time has positive effects.
By the way, max- imum amount of resulted extractions total
yield is seven times more than minimum of total yields
extraction. According to be obtained results and old studies,
total yield extraction increased with pressure, which was due to
increase of SC-CO2 density at high pressure. But however, an
increase in the pressure level, led to an unexpected reduction in
the extraction yield. These unexpected results probably can be
related to the reduced diffusion rates of the extracted essence
from the plant matrix to the supercritical uid CO2 . In this way,
also the density of CO2 at constant pressure is reduced with
increasing temperature and leading to reduce the solvent power
of supercritical CO2 . Temperature effect on process of
supercritical extraction is more complex. The isobaric increase of
temperature diminishes the solvent density but also increase the
solute vapor pressure. In critical point vicinity, small change in
temperature produces large variations in solvent density. Supercritical extraction process had more inuence on the yield than
CO2 ow rate. However at higher pressures, the extraction yield
increase with dynamic extraction time and extraction rate is
higher [17,22,41].
3.2. SCE total concentration results
In addition total yields extraction, by doing any test, it is possible to calculate supercritical Fluid density by dividing the amount
grams of menthes essence on the amount grams of supercritical
uid of CO2 . If we can preserve the average of supercritical uid
concentration in maximum in laboratory condition, we can gain
the more herbal essence if we continue the extraction process
and

Table 4
L16 array and the results of SCE total yield of spearmint essential oil.
Run no.

P (bar)

T ( C)

Di. ( m)

QCO2 (ml/s)

D.t. (min)

S.t. (min)

Yield (%)

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16

90
90
90
90
100
100
100
100
140
140
140
140
170
170
170
170

35
40
45
50
35
40
45
50
35
40
45
50
35
40
45
50

250
500
710
1000
500
250
1000
710
710
1000
250
500
1000
710
500
250

1
3
5
8
5
8
1
3
8
5
3
1
3
1
8
5

30
50
90
120
120
90
50
30
50
30
120
90
90
120
30
50

5060
5060
5060
5060
5060
5060
5060
5060
5060
5060
5060
5060
5060
5060
5060
5060

0.48
1.26
1.82
0.61
1.60
0.42
0.79
0.33
0.97
0.43
0.58
0.76
0.37
0.65
0.94
0.25

Table 5
L16 array and the results of SCE total concentration of spearmint essential oil.
Run no.

P (bar)

T ( C)

Di. ( m)

QCO2 (ml/s)

D.t. (min)

S.t. (min)

C (mg/ml)

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16

90
90
90
90
100
100
100
100
140
140
140
140
170
170
170
170

35
40
45
50
35
40
45
50
35
40
45
50
35
40
45
50

250
500
710
1000
500
250
1000
710
710
1000
250
500
1000
710
500
250

1
3
5
8
5
8
1
3
8
5
3
1
3
1
8
5

30
50
90
120
120
90
50
30
50
30
120
90
90
120
30
50

5060
5060
5060
5060
5060
5060
5060
5060
5060
5060
5060
5060
5060
5060
5060
5060

0.033
0.013
0.006
0.001
0.004
0.001
0.023
0.005
0.004
0.004
0.003
0.014
0.002
0.008
0.007
0.002

this question can be considered so important regarding

economical and semi-industrial pilot point of view.
We have the total concentration of supercritical uid of CO2 for
all of the 16 runs of essential oil spearmint supercritical extraction
test in Table 5. It is seen carefully in this table that the runs (4
and
6) have the lowest amount of density (0.001 mg/ml) and the run
no.1 has the maximum amount of concentration (0.033 mg/ml).
The difference of maximum and minimum of amount of density is
thirty three times.
One of the remarkable points in this table is that, with the
decrease of dynamic time and supercritical uids velocity, total
concentration of uid increases and, then it returns to
equilibrium state. In the short times and the low velocity of
supercritical uid of CO2 , the extraction process is not far from of
counterpoise and is near to the saturation. When the time of the
test lasts, the concen- tration of supercritical uid decrease, thus,
the reduction of total
concentration is happened. When the uid ow rate is more
than consumed feed powder in the test, the extraction process is
far from the equilibrium and supercritical uid concentration cant
be high.

the other hand, the more we have the tests, the more we have
the accumulation and augmentation of errors.
To decrease the number of tests and errors in the framework
of Taguchi method or orthogonal designing, the number of tests
was reduced to 16 that considering once repetition the number
of required tests was 32. Based on the result of Table 4, the optimization related to Taguchi method has been done with qualitek4 software, and the maximum yield of supercritical extraction
was calculated. Also, the optimum conditions for achieving the
maxi- mum total yield of supercritical extraction summarized in
Table 6. In optimum laboratory condition, the maximum total
yield of supercritical extraction of menthe herbal essence is
2.032%. with to pay attention to the optimum condition (Table 6)
the pressure is 90 bar and low level, and dynamic time of 120
min to become equal high range amount of dynamic time (Table
2), we could pre- dict that increase in the dynamic time above 120
min, the total yield extraction of M. spicata L. essential oil is
higher.

Table 6
Optimum condition and performance for total SCE yield of spearmint essential oil.

3.3. Optimization the total yield in SCE of spearmint essential oil

Indeed, the pressure and the temperature of extraction, the
mean particles size of feed powder, the ow rate of supercritical
uid, and dynamic time are the most important parameters in the
supercritical extraction being efcient on the nal yield of extraction. In order to examine the effects of these ve factors, each
in four different levels on the yield of extraction, generally we need
one to 1024(45 ) tests that are so costly and need to very long

No.
1
2
3
4
time.
5

Factors
Pressure (bar)
Temperature ( C)
Mean particle Size ( m)
Flow rate (ml/s)
On
Dynamic time (min)

Level

L. description

Static time

1
3
2
3
4

90
45
500
5
120

5060

Expected result (total yield) at optimum condition = % 2.032

Table 7
Optimum condition and performance for total SCE concentration of spearmint
essential oil (S.t. 5060 min).
No.

Factors

Level

L. description

1
2
3
4
5

Pressure (bar)
Temperature ( C)
Mean particle Size ( m)
Flow rate (ml/s)
Dynamic time (min)

1
1
1

90
35
250
1
30

Expected result (total yield) at optimum condition = 0.033 mg/ml

3.4. Optimization the total concentration in SCE of spearmint

essential oil
With the same method of optimization for the total yield of
supercritical extraction, explained in 3-3, doing 16 times of tests
and its repetition, it can optimize supercritical concentration of
CO2 for extracting M. spicata L. and regarding the optimum
condition, the maximum concentration of herbal essence in
supercritical CO2 is calculated. It can observe these results in Table
7. The maximum concentration of supercritical uid CO2 is
0.033 mg/ml. Table 7 shows in the optimum condition for total
essential oil concentra- tion in supercritical uid CO2 at
supercritical extraction process, all parameters of pressure,
temperature, mean particle size, ow
rate and dynamic time are at least or rst level (90 bar, 35 C,
250 m, 1 ml/s, 30 min). with to give a clear description, if the
examination conditions modied and again design of experiments
(DOE) performed, perhaps for decrease of amount of some factors
such as dynamic time, mean particle size, ow rate and pressure, the concentration of essential oil in supercritical uid CO2
increase.

3.5. Retesting in optimal conditions for conrm of results

(conrmation experiment)
As we have remembered, in the framework of Taguchi method
test and (Qualitek-4) software, besides the determination and the
prediction of optimum condition, it is possible to calculate the
max- imum yield of extraction and the maximum of
supercritical total concentration. One of the advantages of this
method is its ability of reexamination and conrmation in the
same optimum and given conditions. On the other hand, in the
case of some contradictions in the results of the conrmatory
tests with the estimation calculated amount or prediction amount
of Qualitek-4 software, it can change and rectify the parameters
of test, specially its levels, in order to attain better results. It
should say that the exibility of this method is so high.
In this research, two tests with the same optimum conditions
determined in Tables 6 and 7, have been done and the results
show that the amounts of 2.032% for optimum amount of
supercritical extractions yield, and 0.033 mg/ml for the total
optimum concen- tration supercritical uid CO2 are authentic.
In other words, in optimum conditions: pressure 90 bar, temperature 45 C, mean particles size of feed powder 500 m,
supercritical uid ow rate 5 ml/s, and dynamic time 120 min,
we have the maximum total yield of supercritical extraction
based on herbal essence of spearmint that is equal to 2.032%.
Also, considering the optimum conditions: pressure 90 bar, temperature 35

C,
mean particles size of feed power 250 m, supercritical uid ow
rate of 1 ml/s and dynamic time of 30 min, the maximum amount
of optimum supercritical uid concentration is equal to 0.033
mg/ml which is practically authentic. In this laboratory condition,
super- critical uids concentration has the maximum amount.

3.6. Comparison results and old researches

Before this research, experiment designing and optimization of
the supercritical extraction of the spearmint edible herbal
essence has been done in different manner like: (A) full factorial
complete randomized design (FFCRD) with Minitab V.14
[17,25,36], (B) one- way analysis variance with Software statistic a
(Stat Soft Inc. USA) [43], (C) Response surface methodology
(RSM) and face-centered central composite design (FC-CCD)
with software Minitab V.14. (Minitab Inc., PA, USA) [27].
The use of Taguchi method for experiment designing and optimization of supercritical extraction has not been reported so far
and it seems to be for the rst time. Simultaneously, in this
study, we have considered the effect of ve different and
important parame- ters on supercritical extraction each one in four
levels, while in the other and old studies, we had studied the effect
of three parameters in three levels. Although in earlier researches
or in the other manner of optimization, the maximum amount of
supercritical extractions yield had been reported more or the
same as this research, but according to results of reexaminations
in optimum condition and the conrmation of predicted and
calculated results, the obtained amounts in the framework of
Taguchi method are more reliable and have empirical and
scientic authenticity. On the other hand; the examination and
the optimization of supercritical uid CO2 concentration in supercritical extraction process has been reported
for
the rst time in this study.
4. Conclusion
Applying Taguchi method for the rst time, in this laboratorial research, supercritical extraction of edible spearmint herbal
essence was done in order to achieve the maximum of extractions
total yield and supercritical uid concentration. We have determined effect of ve efcient parameters in the process of
extraction regarding optimum conditions. The optimum
conditions of extrac- tion for the maximum yield of extraction
are 2.032%: pressure
of 90 bar, the temperature of 45 C, medium size of feed powder
500 m, supercritical uid ow of 5 ml/s, dynamic time of 120 min
and in order to access the maximum concentration (0.033 mg/ml)
of supercritical uid of CO2 are pressure of 90 bar, temperature of
35 C, medium range of feed powder 250 m, supercritical ow
rate 1 ml/s and dynamic time 30 min. In other words, according
to predicted and calculated results that have been conrmed in
reexaminations with optimal conditions, and based on supercritical extraction, it can extract more than 2 kg of valuable and costly
essential oil from each 100 kg of dried shrubs of spearmint.
Acknowledgment
Hereby we appreciate all those who cooperated with us at different stages of this experimental research and study, particularly
library and laboratory staff of Sharif, Kashan and Shiraz University, as well as the great helps of Gachsaran Gas and Oil Production
Company (GS NISOC).
Appendix A. Supplementary data
Supplementary data associated with this article can be found,
in the online version, at doi:10.1016/j.supu.2012.03.011.
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