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

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

Article history:
In this study, the essential oil of aerial parts species of a plant called Spearmint was extracted by CO2
Received 15 February 2012
to optimize the results of the supercritical extraction process. In order to achieve maximum total
Received in revised form 18 March 2012
Accepted 19 March 2012 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.
Keywords:
The opti-
mizing conditions for SCE and SF-CO2 concentration are: 90 bar, 45 C, 500 m, 5 ml/s, 120 min and 90
bar,
Supercritical extraction
35 C, 250 m, 1 ml/s, 30 min. SCE total yield and concentration are 2.032% and 0.033 mg/ml. In sim-
Spearmint
CO2
Total yield ilar and optimal conditions of extraction, supplementary tests were done so resulted and experimental
Essential oil and Taguchi method 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 a cross breeding species resulted from M. longifolia and rotundifo-


lia. Menthe is one of the herbs that are so important because of its
Lamiaceae family is one of the biggest herbal families that essence and pharmaceutical compounds.
have geographical and universal dispersion. This type is grown Because of its benets and economical values the menthe is one
in all around the world apart from Antarctic and northern region. of the most important herb and many researchers have focused
These families consist of about 200 genuses and between two or their attention on it in the way they can increase its yield and
5000 species of fragrant plant and short shrubs having a large they can know the important factors involving the quality and the
diversity [13]. Respectively, morphological characteristics of Lam quan- tity of the extraction essence. The green menthe or the
families includes quadrangle stems, simple leaves arranged in edible one that is the other species of this family, has more usage
vertical direc- tion that have the reciprocal and alternative [7,913].This family and this herbal species of mint is the rich
forms, with ve-part sepal and two or four stamens ocymun source of poly phenolic compounds and, therefore have a strong
bacilicum, rosmarinum ofcinalis, Thymus vulgaris, Origanum antioxidant prop- erties used in alimentation industry that
vulgare and Menthe belong to the same family [4]. prevent the oxidation. This material can be a suitable substitute
The menthe or mentha genus, that has a vast diversity and for articial and industrial antioxidants like butylated hydroxyl
importance, is one of the most important one in this family used toluene (BHT), and butylated hydroxyl anisole(BHA). Because of
by human being over 2000 years that is grown in all of 5 conti- the harmful effect of articial and industrial antioxidants on
nents having remarkable and commercial importance. Menthe human beings health; the natural alternative is more
has thirteen hybrid named and composed of 19 geographical emphasized [1416].
diversity [57]. This menthe genus has 2530 particular species The recent studies show that the Lamiaceae is a potential
growing in Europe, Asia, and South Africa [8]. Some of the resource for natural antioxidants. This property depends on some
important species of this genus are Mentha spicata L., piperita L., compounds such as avonoids, diterpenes, phenolic and phenolic
aquatica L., pulegium L., etc. that grow in all around the world. The acid [1722]. The antimicrobial property of both species of menthe
M. spicata L. or spearmint species or with the abbreviated name herbal essence has been proven and has a vast industrial, phar-
mint is the same green men- the or edible menthe that is used maceutical, and cosmetic use and its consumption is increasing in
more frequently. Spearmint is the world [2327]. The menthe is largely used in the conventional
medicine because of its antispasmodic, antiseptic, and emmena-
gogue effects. The spearmint essential oil is used for the different
Corresponding author. Tel.: +98 7429122461; fax: +98 7422224015.
kind of chewing gums, beverages, cosmetic products, perfumery,
E-mail address: ansari karamat2006@yahoo.com (K. Ansari).
toothpastes and mouth washes [28,29].

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


vone, 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 proper-
ties, 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 Fig. 1. Plant genus of M. spicata L (spearmint).
high temperature, are not suitable for the valuable compounds
that are sensitive to tem- Table 1
perature and can be resulted in reduction or loss of product.
The supercritical extraction methods that is completely new than Average diameter particles size and weight samples.

these methods has not the problems of earlier and conventional Sample no. Mean particle size ( m) Weight (g)
techniques. Because of its unique properties such as high sol-
1 250 164.15
ubility, low viscosity of supercritical uid and ne selectivity 2 500 263.33
property that present a vast use in alimentation and 3 710 105.42
pharmaceuti- cal industries and the extraction of natural and 4 1000 111.08
herbal raw materials [3439]. Sum 643.98
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 super- 2.1. Sample preparation
critical uids has no free residue in product having
environmentally friendly character that is highly suitable for We have grinded the dried herbal samples of spearmint near
the supercritical extraction. Despite of multiple problems and to the period of tests performance in one suitable mill carefully
because of the simple and facility of use, the manner of and with patience in a way that the temperature of the samples
extraction with sol- vent is common yet [40,41]. So far the in order to preventing the increase of temperature. Each sort of
extraction of the herbal essence of M. spicata L. has been temperatures increase of the samples results reduce of quantity
reported in conventional common manner such as and quality of extractive essential oil. We have prepared the
distillation and the extraction with different chemical powder using suitable sieves and shaking machines in four sizes
solvents and in some cases the supercrit- ical extraction of and the granulated samples were stored in 4 glass container. In
herbal essence has been included. In some cases of the Table 1, the mean particles size of samples and the amount of
supercritical extraction, the co-solvent or mod- iers (co- their weight have obtained. To regulate the time of grinding, it can
solvent) has been used for increasing the yield of extraction achieve the approximated equality of weight in samples that is
[17,23,25,27,36,4245]. so important for all the tests.
In some published articles and most review papers related
to supercritical extraction of the different herbal species in 2.2. Material and reagent
which, the extraction of herbal essence of M. spicata L. has
been dis- cussed using mathematical modeling, the extraction of Some of the consumable materials are necessary for all the
edible an herbal menthe has been also studied [19,24,45]. In supercritical tests. Carbon dioxides storages with 99.9% of purity
some of pub- lished articles related to the supercritical have been prepared in Rohams factory. Sealed dark bottles with
extraction of this herbal species of spearmint, the design of caps in little volume (1 ml) for preserving products essence, spe-
experiment has been done and we have calculated the maximum cial tissues for the sacs in which the powder of tests have been
yield of the herbal essence and the optimal condition of the reserved, chemical materials and required solvents such as ethanol
extraction. In spite of the fact that researches done in this area, and acetone all have been purchased from the market.
it seems optimization of supercriti- cal extraction of spearmint
essential oil has been performed using Taguchi testing method 2.3. Design of experiments
was not done yet and this is the rst report announcing
[17,25,27,36]. Fig. 1 shows this valuable herbal plant. 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
2. Materials and method
to reduce the number of tests, it is necessary to design of the
experiment (DOE). In general, DOE can reduce the costs and
Herbal samples of M. spicata L. were cut and collected as the
economize the time. Also, DOE can be done in different manners.
aerial part and near to the period of owering at the end of
Generally In the process of the supercritical extraction of the
summer from the farms of south west of Iran in Kohgiluye and
herbal essence, the most important factors concerning the total
Buyerahmad province. The samples were very fresh and fragrant
yield of extraction and the concentration of the essence in the
and dried in shadow along with dry weather with low moisture
supercritical uid are: pres- sure and the temperature of the
and have been kept in suitable container and lace before the
extraction, mean particles size of feed powder, ow rate or the
beginning of the tests. The time of drying lasted about 2 weeks.
velocity of supercritical uid and the time of the test or dynamic
The wet samples had more than 80 percent humidity.
time.
Table 2
different units. In this research, the concentration of essential oil
Variable parameter range in SCE for spearmint essential oil.
in supercritical uid CO2 has been expressed as g/L or mg/ml. To
No. Parameters Unit Variable range
achieve the maximum yield of extraction process and maximum
1 Pressure (P) bar 90 < P < 300 density of supercritical uid are the most important goal. The total

2 Temperature (T) C 35 < T < 60 yield resulted number that is in fraction and percent is very little
3 Mean particle size (Di.) m 250 < Di. < 5000
and can be dened as mg/g (milligrams of extractive essence
4 Flow rate (QCO2 ) ml/s 1 < QCO2 <
5 Dynamic time (D.t.) min 30 < D.t. < 120 divide on gram consumed food) [17,25,4951].

Table 3
Experimental levels of the factors used in Taguchi method for SCE of spearmint
2.5. SCE process optimization and statically analysis
essential oil.
Generally, in optimization, it came to be considered the
Factors (parameters) Factors levels achieve- ment of goal with nearest, best and cheapest route or
1 2 3 4 maximum
and minimum of one special product in the process. For the
harmful minimum range of production or its deletion was the
products,
Pressure (bar) 90 100 140 170
Temperature ( C) 35 40 45 50
rst goal. In order to produce valuable and expensive materials,
Mean particle Size ( m) 250 500 710 1000 maxi-
mum range of production with low cost has been the rst regard.
Flow rate (ml/s) 1 3 5 8 In clinical researches such as supercritical extraction in which the

Dynamic time (min) 30 50 90 120


costs of tests are high, regarding the high pressure that is out of
criti- cal domain and about which the safety considerations are
Almost in the all of former researches on the supercritical exclusive and some of the special tests whose performance
extrac- tion and such as researches done on the spearmint demand much attention, it is necessary to apply all of the safety
essential oil supercritical extraction, we have used the same ve considerations in order to optimize the process of this tests that
parameters or some of them. Of course for this special purpose, may be in some cases the only choice and issue. The
most of stud- ies have been done based on three parametric tests. optimization can be done in different manners like the design of
According to the researches on this herbal species with the experiment. In some cases such as complete randomized design
characteristics of the our laboratorial pilot and estimate/error test (CRD) and response surface methodology (RSM), when we have
in different condi- tions, we have determined the domain of the tests with multiple param- eters, the calculations will be
parameters changes, that summarized in Table 2 [17,25,27,36,43]. more complex and time-consuming [17,25,36,52,53].
In this experimental research, in order to design of the exper- The use of Taguchi method in order to design and optimize the
iment, we have used Taguchi method because of facility and supercritical tests besides simplicity has reliable, good and exact
simplicity of use. This method is so precise and reliable, having results. According to the parameters and the levels of change (5
different use in science and engineering. When the number of factors, each one in 4 level) in this manner we have as below:
ef- cient parameters is more than three, the use of Taguchi In order to carry out the tests, we have used orthogonal array
method is suitable for designing the tests because of economy in (L16).Every time, we change laboratory conditions, considering
time, cost and depreciation of laboratory devices. Furthermore, six- teen times of test we can calculate total yield of extraction
the result of this method, highly depend on experience and process and supercritical uids concentration. To avoid different
further test, ensur- ing better result. Also, if necessary, it can errors, it can repeat each run two or several times.
repeat the test in different array and conditions in order to get
better results [4648].
(1) An optimum condition of supercritical extraction is determined
According to this research, in Table 3, the efcient factors on
for achieving maximum yield of extraction and supercritical
quantity and quality of extractive essence in supercritical man-
uids concentration.
ner and total yield of extraction process consist of: pressure and
(2) Optimum response or nal yield of extraction and
temperature of the extractions utensil, mean particles size of feed
supercritical uids concentration are estimated and
powder, ow rate of supercritical period, and dynamic time, each
calculated.
one is seen in four different levels. L16 array has been prepared
(3) To ensure the calculated and assessed results with the same
based on this table.
optimal conditions in item no. 2, we have done conrmation
experiment.
2.4. Total yield extraction and supercritical uid concentration

Total yield of supercritical extraction has been dened in dif- The determination of optimum conditions and the optimiza-
ferent manners. In researches works that most insist more on tion of supercritical extraction and the calculation of maximum
extractions modeling and equilibrium state, the yield is calculated response in total yields extraction and supercritical uid concen-
by dividing the amount mass of extractive essence in supercritical tration have been done with use of qualitek-4 software and the
process on maximum mass of extractable essence or the amount analysis of variance or ANOVA (items no. 2 and 3). Also, it can
of primary mass in feed powder. Any report in which determine the effect of different parameters on optimum response
experimental aspects of process and the optimization of of extraction. The use of this software and ANOVA provides the
supercritical extrac- tion are discussed, denes total yield with conditions in which it is possible to reexamine or conrmation
dividing the mass of extractive essence on the mass of consumed experiment and become aware of the accuracy of optimum
feed. In both of these manners, this ratio is expressed as amount.
fractional or percent number. In this study we have used second In the case of contradiction with estimated and calculated
manner and the yield is dened as the gram amount of produced amounts and in order to reduce or delete the errors, it can change
essence on the gram amount of consumed feed powder. the conditions of the test, in other words it is possible to change
The concentration of supercritical uid that shows the amount the levels of parameters or revise the test and get to better
of dissolved herbal essence in the supercritical uid in different results. On the other hand, this test presents (Taguchi method)
laboratory conditions has always the same criteria and can just different advantages that help to identify and delete the most
have efcient ones; it is an important subject [54,55].
2.6. Static and dynamic times in uids circulation pumps, temperature control system (TIC),
SCE electric heater of water

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 multi-
purpose 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 pres-
sure tolerance arrives up to 400 bar and through a good control;
operation of the supercritical extraction can be performed in tem-
perature 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
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 nec-
essary to test it for preventing CO2 leakage. To remove eventual
leakages, it should increase step by step interior pressure of sys-
tem 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 temper-
ature of extractor arrives at 35 C, the needle valve is closed on
the exterior of extractions extractor, this operation increase grad-
ually 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 pres-
sure 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 other words if we increase the pressure, the yield decreases,
super- critical ow rate has no effect as well, after that the factor
Based on experiments designing in the framework of Taguchi of mean particles size of powder has positive effect resulting the
method and according to array and orthogonal L16, and increase of yield, in the case of increased feed powder diameter.
considering the change of condition and different parameters Both last parameters; temperature and dynamic time have an
such as pressure and time of extraction, supercritical ow rate, equal effect on supercritical extractions total yield while heat
mean particles size of feed powder and dynamic time, sixteen temperature has negative and dynamic time has positive effects.
times of test and just one repetition, we have done in sum 32 By the way, max- imum amount of resulted extractions total
main tests. We have sub- sequent results in this section. The yield is seven times more than minimum of total yields
study of different parameters, similar pressure, temperature, extraction. According to be obtained results and old studies,
dynamic time, etc. on supercritical extraction essential oil of M. total yield extraction increased with pressure, which was due to
spicata L. (Spearmint) can be reported by another paper that will increase of SC-CO2 density at high pressure. But however, an
publish. increase in the pressure level, led to an unexpected reduction in
the extraction yield. These unexpected results probably can be
3.1. SCE total yield results related to the reduced diffusion rates of the extracted essence
from the plant matrix to the supercritical uid CO2 . In this way,
At the end of each run, the total yield of supercritical extrac- also the density of CO2 at constant pressure is reduced with
tion is resulted by dividing the amount of produced essence mass increasing temperature and leading to reduce the solvent power
on the mass of consumed feed powder of spearmint herbal of supercritical CO2 . Temperature effect on process of
species that is expressed as a percentile, summarized in Table 4. supercritical extraction is more complex. The isobaric increase of
As we can see in this table, in the case of change in conditions, the temperature diminishes the solvent density but also increase the
amount of total yield extraction is different and is varied of 0.25 solute vapor pressure. In critical point vicinity, small change in
1.82%. The run number (3) has the highest yield and in run 16 or temperature produces large variations in solvent density. Super-
the last run, we have the lowest total yield of extraction. In both critical extraction process had more inuence on the yield than
of the runs, just the parameter of supercritical ow rate is xing CO2 ow rate. However at higher pressures, the extraction yield
and the other factors of test conditions are different. Among the increase with dynamic extraction time and extraction rate is
different factors or conditions of these two runs with the higher [17,22,41].
minimum or maximum nal yield, the pressure has the most
difference and the heat tem- perature and dynamic time each one
3.2. SCE total concentration results
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
In addition total yields extraction, by doing any test, it is possi-
test, the static mode time is xed on
ble to calculate supercritical Fluid density by dividing the amount
5060 min or 1 h. Addition of a static step in to the extraction
grams of menthes essence on the amount grams of supercritical
con- dition did not have any effect on the recovery of essence oil
uid of CO2 . If we can preserve the average of supercritical uid
from plant genus of M. spicata L.
concentration in maximum in laboratory condition, we can gain
On the other hand, it can say that the parameter of pressure
the more herbal essence if we continue the extraction process
has the most negative effect in the yield of supercritical
and
extraction, in
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 90 35 250 1 30 5060 0.48


2 90 40 500 3 50 5060 1.26
3 90 45 710 5 90 5060 1.82
4 90 50 1000 8 120 5060 0.61
5 100 35 500 5 120 5060 1.60
6 100 40 250 8 90 5060 0.42
7 100 45 1000 1 50 5060 0.79
8 100 50 710 3 30 5060 0.33
9 140 35 710 8 50 5060 0.97
10 140 40 1000 5 30 5060 0.43
11 140 45 250 3 120 5060 0.58
12 140 50 500 1 90 5060 0.76
13 170 35 1000 3 90 5060 0.37
14 170 40 710 1 120 5060 0.65
15 170 45 500 8 30 5060 0.94
16 170 50 250 5 50 5060 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 90 35 250 1 30 5060 0.033


2 90 40 500 3 50 5060 0.013
3 90 45 710 5 90 5060 0.006
4 90 50 1000 8 120 5060 0.001
5 100 35 500 5 120 5060 0.004
6 100 40 250 8 90 5060 0.001
7 100 45 1000 1 50 5060 0.023
8 100 50 710 3 30 5060 0.005
9 140 35 710 8 50 5060 0.004
10 140 40 1000 5 30 5060 0.004
11 140 45 250 3 120 5060 0.003
12 140 50 500 1 90 5060 0.014
13 170 35 1000 3 90 5060 0.002
14 170 40 710 1 120 5060 0.008
15 170 45 500 8 30 5060 0.007
16 170 50 250 5 50 5060 0.002

this question can be considered so important regarding the other hand, the more we have the tests, the more we have
economical and semi-industrial pilot point of view. the accumulation and augmentation of errors.
We have the total concentration of supercritical uid of CO2 for To decrease the number of tests and errors in the framework
all of the 16 runs of essential oil spearmint supercritical extraction of Taguchi method or orthogonal designing, the number of tests
test in Table 5. It is seen carefully in this table that the runs (4 was reduced to 16 that considering once repetition the number
and of required tests was 32. Based on the result of Table 4, the opti-
6) have the lowest amount of density (0.001 mg/ml) and the run mization related to Taguchi method has been done with qualitek-
no.1 has the maximum amount of concentration (0.033 mg/ml). 4 software, and the maximum yield of supercritical extraction
The difference of maximum and minimum of amount of density is was calculated. Also, the optimum conditions for achieving the
thirty three times. maxi- mum total yield of supercritical extraction summarized in
One of the remarkable points in this table is that, with the Table 6. In optimum laboratory condition, the maximum total
decrease of dynamic time and supercritical uids velocity, total yield of supercritical extraction of menthe herbal essence is
concentration of uid increases and, then it returns to 2.032%. with to pay attention to the optimum condition (Table 6)
equilibrium state. In the short times and the low velocity of the pressure is 90 bar and low level, and dynamic time of 120
supercritical uid of CO2 , the extraction process is not far from of min to become equal high range amount of dynamic time (Table
counterpoise and is near to the saturation. When the time of the 2), we could pre- dict that increase in the dynamic time above 120
test lasts, the concen- tration of supercritical uid decrease, thus, min, the total yield extraction of M. spicata L. essential oil is
the reduction of total higher.
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.
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 No. Factors Level L. description Static time

mean particles size of feed powder, the ow rate of supercritical 1 Pressure (bar) 1 90 5060
uid, and dynamic time are the most important parameters in the 2 Temperature ( C) 3 45
3 Mean particle Size ( m) 2 500
supercritical extraction being efcient on the nal yield of extrac-
4 Flow rate (ml/s) 3 5
tion. In order to examine the effects of these ve factors, each time. On
5 Dynamic time (min) 4 120
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
Expected result (total yield) at optimum condition = % 2.032
Table 7
3.6. Comparison results and old researches
Optimum condition and performance for total SCE concentration of spearmint
essential oil (S.t. 5060 min).
Before this research, experiment designing and optimization of
No. Factors Level L. description
the supercritical extraction of the spearmint edible herbal
1 Pressure (bar) 1 90 essence has been done in different manner like: (A) full factorial
2 Temperature ( C) 1 35 complete randomized design (FFCRD) with Minitab V.14
3 Mean particle Size ( m) 1 250
4 Flow rate (ml/s) 1
[17,25,36], (B) one- way analysis variance with Software statistic a
5 Dynamic time (min) 1 30 (Stat Soft Inc. USA) [43], (C) Response surface methodology
(RSM) and face-centered central composite design (FC-CCD)
Expected result (total yield) at optimum condition = 0.033 mg/ml
with software Minitab V.14. (Minitab Inc., PA, USA) [27].
The use of Taguchi method for experiment designing and opti-
mization of supercritical extraction has not been reported so far
and it seems to be for the rst time. Simultaneously, in this
3.4. Optimization the total concentration in SCE of spearmint study, we have considered the effect of ve different and
essential oil important parame- ters on supercritical extraction each one in four
levels, while in the other and old studies, we had studied the effect
With the same method of optimization for the total yield of of three parameters in three levels. Although in earlier researches
supercritical extraction, explained in 3-3, doing 16 times of tests or in the other manner of optimization, the maximum amount of
and its repetition, it can optimize supercritical concentration of
supercritical extractions yield had been reported more or the
CO2 for extracting M. spicata L. and regarding the optimum
same as this research, but according to results of reexaminations
condition, the maximum concentration of herbal essence in in optimum condition and the conrmation of predicted and
supercritical CO2 is calculated. It can observe these results in Table calculated results, the obtained amounts in the framework of
7. The maximum concentration of supercritical uid CO2 is Taguchi method are more reliable and have empirical and
0.033 mg/ml. Table 7 shows in the optimum condition for total scientic authenticity. On the other hand; the examination and
essential oil concentra- tion in supercritical uid CO2 at the optimization of supercritical uid CO2 con-
supercritical extraction process, all parameters of pressure, centration in supercritical extraction process has been reported
temperature, mean particle size, ow for
rate and dynamic time are at least or rst level (90 bar, 35 C, the rst time in this study.
250 m, 1 ml/s, 30 min). with to give a clear description, if the
examination conditions modied and again design of experiments 4. Conclusion
(DOE) performed, perhaps for decrease of amount of some factors
such as dynamic time, mean particle size, ow rate and pres- Applying Taguchi method for the rst time, in this laborato-
sure, the concentration of essential oil in supercritical uid CO2 rial research, supercritical extraction of edible spearmint herbal
increase. essence was done in order to achieve the maximum of extractions
total yield and supercritical uid concentration. We have deter-
mined effect of ve efcient parameters in the process of
3.5. Retesting in optimal conditions for conrm of results extraction regarding optimum conditions. The optimum
(conrmation experiment) conditions of extrac- tion for the maximum yield of extraction
are 2.032%: pressure
As we have remembered, in the framework of Taguchi method of 90 bar, the temperature of 45 C, medium size of feed powder
test and (Qualitek-4) software, besides the determination and the 500 m, supercritical uid ow of 5 ml/s, dynamic time of 120 min
prediction of optimum condition, it is possible to calculate the and in order to access the maximum concentration (0.033 mg/ml)
max- imum yield of extraction and the maximum of of supercritical uid of CO2 are pressure of 90 bar, temperature of
supercritical total concentration. One of the advantages of this 35 C, medium range of feed powder 250 m, supercritical ow
method is its ability of reexamination and conrmation in the rate 1 ml/s and dynamic time 30 min. In other words, according
same optimum and given conditions. On the other hand, in the to predicted and calculated results that have been conrmed in
case of some contradictions in the results of the conrmatory reexaminations with optimal conditions, and based on supercriti-
tests with the estimation calculated amount or prediction amount cal extraction, it can extract more than 2 kg of valuable and costly
of Qualitek-4 software, it can change and rectify the parameters essential oil from each 100 kg of dried shrubs of spearmint.
of test, specially its levels, in order to attain better results. It
should say that the exibility of this method is so high. Acknowledgment
In this research, two tests with the same optimum conditions
determined in Tables 6 and 7, have been done and the results Hereby we appreciate all those who cooperated with us at dif-
show that the amounts of 2.032% for optimum amount of ferent stages of this experimental research and study, particularly
supercritical extractions yield, and 0.033 mg/ml for the total library and laboratory staff of Sharif, Kashan and Shiraz Univer-
optimum concen- tration supercritical uid CO2 are authentic. sity, as well as the great helps of Gachsaran Gas and Oil Production
In other words, in optimum conditions: pressure 90 bar, tem- Company (GS NISOC).
perature 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 Appendix A. Supplementary data
based on herbal essence of spearmint that is equal to 2.032%.
Also, consid- Supplementary data associated with this article can be found,
ering the optimum conditions: pressure 90 bar, temperature 35 in the online version, at doi:10.1016/j.supu.2012.03.011.

C,
mean particles size of feed power 250 m, supercritical uid ow
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