Industrial Crops and Products 76 (2015) 809816

Contents lists available at ScienceDirect

Industrial Crops and Products

journa l homepage: www.e lsev ier .com/ locate / indcrop

Chemical composition and yield of essential oils f(Sature tr

Sayadeh louMohamma Flavarjan Brab Shahrekord B ry, DeShahrekord, Ira

a r t i c l

Article history:Received 1 April 2015Received in revised form 26 July 2015Accepted 28 July 2015

Keywords:Essential oilChemical comMicrowave-asInnovative tecMicrowave-asSatureja bachti

ic herdetermine the effect of extraction methods on qualitative and quantitative characteristics of the essentialoil of Bakhtiari savory was extracted by different methods. Conventional methods [hydrodistillation bytwo Clevenger-type apparatus: British Pharmacopeia (HDBP) and Research Institute of Forests and Range-lands, Iran (HDRIFR), and traditional steam and water distillation (TSWD)], an innovative technique steamdistillation (SDinnov), microwave-assisted steam hydro-diffusion (MSHD 400 and 800 W), and microwave-assisted hydro-diffusion (MHD 400 and 800 W) were used to extract essential oil from the aerial parts of

1. Introdu

The genaceae contawidely distSatureja L. isthe mount2008). A toora of IraBurge and S(Ghasemi-PMarzeh-e-quently use

CorresponE-mail add

(A. Ghasemi Pi

http://dx.doi.o0926-6690/ positionsisted hydro-diffusionhnique steam distillationsisted steam hydro-diffusionarica

savory and their results were compared. The essential oils of all samples were analyzed using GCFID,GC/MS, and for comparison, a sample was analyzed using head space solid-phase microextraction (HS-SPME). The highest essential oil yields were obtained from two methods, including HDBP, and MSHD400 W. Signicant differences occurred among the major constituents in essential oils from extractionmethods, including thymol, carvacrol, -cymene, and -terpinene. The highest amounts of thymol andcarvacrol were obtained from MHD (800 W) method in comparison other methods. The highest content ofmonoterpene hydrocarbons was obtained from HS-SPME, whereas the highest percentage of oxygenatedmonoterpenes was achieved from MHD (800 W) method. In conclusion, extraction of the essential oil fromBakhtiari savory with microwave-assisted hydro-diffusion (MHD) was better in terms of energy saving,extraction time, oxygenated fraction (thymol and carvacrol), and product quality.

2015 Elsevier B.V. All rights reserved.

ction

us Satureja L. (savory) belonging to the family Lami-ins 200 species of herbs and shrubs, often aromatic,ributed in the Mediterranean area (Cronquist, 1988).

represented by 14 species commonly observed amongains in the southwestern part of Iran (Mozaffarian,tal of nine Satureja species have been reported inn (Mozaffarian, 2008), two of which, S. bachtiarica. kallarica Jamzad are distributed in southwestern Iranirbalouti et al., 2014a). S. bachtiarica (Persian name:Koohi) is well-known aromatic plant which is fre-d as spices and as traditional medicinal herbs in Iran. S.

ding author. Fax: +98 381 336 1012.resses: ghasemi@iaushk.ac.ir, aghasemipir@psis.umass.edurbalouti).

bachtiarica is a perennial herb, which it distributed in Zagros moun-tain range, southwestern Iran (Ghasemi-Pirbalouti et al., 2013a).Infusions and decoctions of aerial parts of S. bachtiarica are used forthe treatment of colds, and also are used as analgesic and antisep-tic by the indigenous people of Bakhtiar va Chaharmahal province,southwestern Iran (Ghasemi-Pirbalouti et al., 2014b). S. bachtiar-ica contains various biologically active components, especiallyvolatile compounds and monoterpenes (Sedkon and Jamzad,2000). The essential oil isolated from the aerial parts of S. bachtiaricahas been shown to have biological and pharmacological activi-ties, including antibacterial (Ghasemi-Pirbalouti and Dadfar, 2013;Ghasemi-Pirbalouti et al., 2014b), antifungal (Ghasemi-Pirbaloutiet al., 2011a,b), antioxidant (Ghasemi-Pirbalouti et al., 2014c), andimmunomodulatory effects (Ghasemi-Pirbalouti et al., 2011c).

The conventional methods for the essential oil extraction ofherb materials, including hydrodistillation and steam distillationhave some disadvantages. Losses of some volatile constitutes, lowextraction efciency, degradation of unsaturated or ester com-

rg/10.1016/j.indcrop.2015.07.0682015 Elsevier B.V. All rights reserved.ja bachtiarica Bunge.) under different ex

Mansoreh Memarzadeha, Abdollah Ghasemi Pirbaad AdibNejada

nch, Islamic Azad University, Department of Essential Oil Chemistry, Isfahan, Iranranch, Islamic Azad University, Research Center for Medicinal Plants & EthnoVeterinan

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

Bakhtiari savory is a perennial aromatrom Bakhtiari savoryaction methods

tib,,

partment of Medicinal Plants, PO Box: 166,

b distributed in Zagros mountain range, southwestern Iran. To

810 S.M. Memarzadeh et al. / Industrial Crops and Products 76 (2015) 809816

pounds through thermal or hydrolytic effects and toxic solventresidue in the extract may be encountered using these extrac-tion methods (Tigrine-Kordjani et al., 2006; Prino-Issartier et al.,2013). In addition, these methods have distinct drawbacks suchas time-conlarge volumopment of drugs indusenvironmenfor minimizdemand to demand thproducts (Tthe extractithe oil yieldsuch as miction, and u(Kaufmannmicrowaveas microwahydrodiffusmicrowavedistillation et al., 2006)2007) beingof advantagequipment via mild incof which co(Prino-Issamicrowavecesses and used in che(Stashenkoand Rezaeiutilized a mnents from knowledge of savory spthe ability ooils from Band the mooil quality,

2. Materia

2.1. Plant m

The aeriabachtiarica southwestelevel in Julyand voucheium of ReseChaharmahwere dried dark conditfood procespieces of de

2.2. Chemic

Homoloheim, Germcalculation

(Na2SO4) for drying extracted oil was purchased from Merck Co.(Darmstadt, Germany).

2.3. Extraction procedures

exted

ds, thxtraial oi

Hydr00 g oleven

(FigDRIFno my antion w

Tradi aeril ste

steaal betus (n. Thulph

Steam essted bsign

steaal betus ws driis. Ea

Microtus aomesed foen soask ccrowe theed baterpara0 W

extrlecte

interied

vialsrform

Micro00 g for 3

The enseted eand 8nt tosuming and labor intensive operations, handling ofes of hazardous solvents (Khajeh, 2011). The devel-new extraction techniques for the food and herbaltries has received a lot of attention lately due to thetal restrictions, reducing waste water, and the needing the energy costs. In addition, there is a constant

improve the quality of essential oils because consumersis quality in their food, pharmaceutical or perfumeryigrine-Kordjani et al., 2006). However, in order to reduceon time, and the operation costs and possibly improve

and the quality of the essential oil, new approachesrowave-assisted extraction, supercritical uid extrac-ltrasound-assisted extraction have also been sought

and Christen, 2002). Driven by these goals, advances in extraction have resulted in a number of techniques suchve steam distillation (Farhat et al., 2011), microwaveion and gravity (Vian et al., 2008; Bousbia et al., 2009),

steam diffusion (Farhat et al., 2009), microwave dryor microwave accelerated distillation (Tigrine-Kordjani

and solvent free microwave extraction (Lucchesi et al., used. Microwave-mediated processes bring a numberes to essential oil extraction thanks to their reducedsize, ease-of-use, speed, ability to control a processrements in heating and low solvent consumption, allntribute to reducing environmental impact and costsrtier et al., 2013). Over the years procedures based on

extraction have replaced some of the conventional pro-other thermal extraction techniques that have beenmical laboratories for decades. Some recently studies

et al., 2004; Tigrine-Kordjani et al., 2006; Golmakani, 2008; Prino-Issartier et al., 2013) have successfully

icrowave oven for the extraction of active compo-medicinal plants/herbs. However, to the best of authorsno work has been published on the microwave-assistedecies. In this study, we present a comparative study off a number of different methods to extract the essentialkhtiari savory (Satureja bachtiarica Bunge.) in order tost advantageous in term of extraction kinetics, essentialand quantity.

l and methods

aterial

l parts (up to 5 cm, 100 g) of Bakhtiari savory (SaturejaBunge.) were collected from Chaharmahal va Bakhtiari,rn Iran (3217N and 5033E) about 2200 m above sea

2012. Plant identities were conrmed by Dr. Shrimardir specimen (no. 1999) has been placed in the Herbar-arch Center for Agricultural and Natural Resources ofal va Bakhtiari, Iran. The fresh aerial of S. bachtiaricainside for ve days at room temperature (30 5 C) inions, and the ground to ne a powder using Endecottsor and passed through a 16 mesh sieve to remove largebris.

als

gous series of C5 C24 n-alkanes (SigmaAldrich, Steine-any) were used for identication of all constituents byof the retention indexes. Anhydrous sodium sulphate

Theperformmethomicroeessent

2.3.1. A 1

two C(HDBP)Iran (H(until dried bextrac

2.3.2. The

ditionaby themateriappara240 midrous s

2.3.3. The

extracthat deby themateriapparaoil waanalys

2.3.4. appara

A dmodihad bea 1 L the mi(outsidproducplant mtype apand 80cient tothe col10 minthen damberwas pe

2.3.5. A 1

water water.a condextracat 400 sufcieraction of the essential oils from S. bachtiarica wasusing different methods, including three conventionalree innovative techniques, and head space solid-phase

ction method. All the results are reported in grams ofls per 100 g of dried savory aerial parts.

odistillation using two Clevenger apparatusf S. bachtiarica was submitted to hydrodistillation usingger-type apparatus, including British Pharmacopeia. 1a) and Research Institute of Forests and Rangelands,R) (Fig. 1b) and extracted with 1 L of water for 240 minore essential oil was obtained). The essential oil washydrous sulphate and stored at 4 C until analysis. Eachas performed at least three times.

tional steam and water distillation procedureal parts of S. bachtiarica (100 g) were submitted to tra-am and water distillation (TSWD). The vapor producedm generator passed through the essential oil rich plantfore being condensed into a receiving Clevenger-typeBritish Pharmacopeia approach) with 1 L of water fore essential oil was obtained, and then dried under anhy-ate. Each extraction was performed at least three times.

distillation apparatus and procedureential oil from the aerial parts of S. bachtiarica wasy an innovative technique steam distillation (SDinnov)ed by Ghasemi Pirbalouti (Fig. 1c). The vapor producedm generator passed through the essential oil rich plantfore being condensed into a receiving Clevenger-typeith 1 L of water for 240 min. The collected essential

ed under anhydrous sulphate and stored at 4 C untilch extraction was performed at least three times.

wave-assisted steam hydro-diffusion (MSHD)nd proceduretic microwave oven (MG-4012 WM/00, LG, Korea) wasr MHD and MSHD operation. 100 g of S. bachtiarica thataked in 1L of distilled water for 30 min were placed inontaining deionized water. The ask was setup withinave oven cavity and a condenser was used on the top

oven) to collect the extracted essential oils. The vapory the microwave passed through the essential oil richial before being condensed into a receiving Clevenger-tus (Fig. 1d). The microwave oven was operated at 400

power level for a period of 30 min. This period was suf-act all the essential oils from the sample. During 30 min,d essential oils were decanted from the condensate inrvals. To remove water, the extracted essential oils wereover anhydrous sodium sulfate, weighed and stored in

at 4 C until they were used for analysis. Each extractioned at least three times.

wave-assisted hydro-diffusion (MHD)of S. bachtiarica that had been soaked in 1 L of distilled0 min were placed in a 1 L ask containing deionizedask was setup within the microwave oven cavity andr was used on the top (outside the oven) to collect thessential oils (Fig. 1d). The microwave oven was operated00 W power level for a period of 30 min. This period was

extract all the essential oils from the sample. During

S.M. Memarzadeh et al. / Industrial Crops and Products 76 (2015) 809816 811

Fig. 1. (a) HDBP, (b) HDRIFR, (c) SDinnov, (d) MSHD and MHD.

812 S.M. Memarzadeh et al. / Industrial Crops and Products 76 (2015) 809816

30 min, the collected essential oils were decanted from the con-densate in 10 min intervals. The essential oil was obtained, driedunder anhydrous sodium sulphate and stored at 4 C until analysis.The extractions were performed at least three times, and the meanvalues were

2.3.6. HeadThe sam

in a coolerdays after czene/carboxcross linkepounds. Thto the headvial) for extinserted intspectrometthe metabo250 C for 6lary column0.25 m lm

2.4. Identi

Chemicagas chromaThe GC ana(Agilent Tector and a acolumn (30phenyl, 95%gas (N2) waprogrammeature was sa ratio split neat.

GCMS performed coupled toquadrupoleCA, USA). A(30 m 0.2tionary phaow rate. T4 C/min raatures werespectra werThe ion sou250 and 15

Chemicabased on thomologoumass spectrand stored iogy) and WpercentageFID respons

2.5. Statisti

The dataSPSS (19.0),and extractcans multip

3. Results and discussion

3.1. Effect of extraction methods on the essential oil yield

esse (S. b, yellsigni1) w

obtaioccusimil37 resper the

Extrith Hicro

me del2011ed th

of gation but 75aveher sultave

, 200ndicae effefect

in ti

fect o

idetion ts shrea. extral, caroundrvacecurshe mtractmicrhe vonvei-Pi

A cohersplanplan

his st diffial oiene (and d, whrvacr

indirol (3 reported.

space solid-phase microextraction (HS-SPME)ples were stored in sealed plastic bags and placed

on ice until analysis by HS-SPME no more than veollection. The SPME device coated with divinyl ben-en/poly dimethyl siloxane (30 m and 20 mm length,

d, bipolar) was used for extraction of volatile com-e ber was extended through the needle and exposed

space above (2 g) of sample contained in a (30 mL glassraction 60 min at 40 C. The ber was then retracted ando the injector of the gas chromatograph coupled to masser equipped with a quadrupole mass analyzer, wherelites were thermally desorbed in the splitless mode at

min, and transferred directly to the analytical capil- (HP-5MS 5% phenyl methyl silicone, 30 m 0.25 mm,

thicknesses).

cation of the oil constituents

l composition of the essential oils was determined bytography (GC) and mass spectrophotometry (GC/MS).lysis was done on an Agilent Technologies 7890 GChnologies, Santa Clara, CA) equipped with a single injec-me ionization detector (FID). An apolar HP-5 capillary

m 0.25 mm, 0.25 m lm thicknesses) coated with 5% methyl polysiloxane were used. The ow of the carriers 0.8 mL/min. Initial column temperature was 60 C andd to increase at 4 C/min to 280 C. The injector temper-et at 280 and 300 C. Split injection was conducted withof 1:40. Samples of the essential oil of 0.1 L were injected

analyses of the samples of the essential oil wereon an Agilent Technologies 7890 gas chromatograph

Agilent 5975C mass selective detector (MSD) and EI mass analyzer (Agilent Technologies, Palo Alto,

HP5MS 5% column (coated with methyl silicone)5 mm, 0.25 m lm thicknesses) was used as the sta-se. Helium was used as the carrier gas at 0.8 mL/minhe temperature was programmed from 60 to 280 C atmp rate. The injector and the GCMS interface temper-

maintained at 290 C and 300 C, respectively. Masse recorded at 70 eV. Mass range was from m/z 50550.rce and the detector temperatures were maintained at0 C, respectively.l compositions of the essential oils were identiedheir retention indices (determined with reference tos series of C5C24 n-alkanes), by comparison of theira with those reported in the literature (Adams, 2007)n NIST 08 (National Institute of Standards and Technol-illey (ChemStation data system) libraries. The peak areas were computed from HP-5 column without the use ofe factors.

cal analyses

was statistically analyzed using one-way ANOVA by and comparison of the means of the main constituentsion yields for the essential oils was evaluated by Dun-le range test (p 0.05).

All savorya clearhad a (p 0.0basis) yields show and 1.times, ence fo2008).that wwith mondaryrapidlyet al., indicatglandsextracin abomicrowat a higrm remicrowRezaeistudy ipositivitive efsaving

3.2. Ef

Theextracponenpeak aferent thymowere fand cabon prwere toils exphase with tusing cGhasem2013).researcof the of the 2015).

In tnicanessent-cymfrom methoand caresultscarvacntial oils extracted from the aerial parts of Bakhtiariachtiarica) by different extraction methods producedow liquid. Results indicated that the extraction methodcant effect on essential oil yield of Bakhtiari savoryith the highest essential oil yields (v/w on dry weightned by HDBP and MSHD 400 W. The lowest essential oilrred by HDRIFR and MHD 400 W (Fig. 2). HDBP and MSHDar oil yield: they reach maximal yields of 1.4 0.010.06 mL/100 g dry plant, in 4 h and 30 min extractionctively. HD is an approved method that is used as refer-

quantication of essential oils (Golmakani and Rezaei,action with MSHD started at much earlier time thanD. This is due to the more efcient heat ow involvedwaves. During microwave assisted extraction of sec-tabolites, such as essential oil from plants, microwaveivers energy to extractant and plant matrix (Zhang). Results of a study by Golmakani and Rezaei (2008)at microwave-assisted hydrodistillation destroyed therden thyme (Thynus vulgaris L.) leaves in 30 min, but they conventional hydrodistillation destroyed the glands

min. Unlike the classical conductive heating methods,s can heat the entire sample almost simultaneously andrate (Kaufmann and Christen, 2002). These results con-s from the literature, which indicate that the use ofs allows extractions to be accelerated (Golmakani and8; Prino-Issartier et al., 2013). In addition, a result thisted other microwave-assisted methods, does not have act on gives a maximum yield (Fig. 2), whereas has a pos-

on extraction kinetics. These results mean a substantialme.

f extraction methods on the essential oil composition

ntities of the extracted essential oils from differentmethods are shown in Table 1 . In total, the 49 com-own in Table 1 consist of about 96100% of total GCThe main components of the essential oils from dif-ction methods were -cymene, -terpinene, linalool,vacrol, and -caryophyllene (Table 1). All essential oils

to be rich in the active monoterpene phenols (thymolrol) and their corresponding monoterpene hydrocar-ors (-cymene and -terpinene). Thymol and carvacrolost abundant components in S. bachtiarica essential

ed using extraction methods expect head space solid-oextraction (HS-SPME) method. This is in agreementalues previously reported for the same species, butntional extraction method (Sedkon and Jamzad, 2000;rbalouti et al., 2013a; Ghasemi-Pirbalouti and Dadfar,mparison of our results with the previous report by

suggests few differences in the volatile compositiont material could be attributed to the geographic origint and to the methods of extraction (Mirhosseini et al.,

tudy, results of analysis of variance indicated that sig-erences occurred among the major constituents in thels from extraction methods. The highest percentages of32.7 0.49%) and -terpinene (23.54 1.09%) obtainedhead space solid-phase microextraction (HS-SPME)ereas the lowest percentages of thymol (6.69 0.31%)ol (4.80 0.08%) obtained from this method. In addition,cated that the percent of thymol (34.53 1.48%) and1.66 0.61%) in the MHD (800 W) method were higher

S.M.

Mem

arzadeh et

al. /

Industrial Crops

and Products

76 (2015)

809816

813

Table 1Effect of different extraction methods on chemical compositions of essential oils from the aerial parts of Bakhtiari savory (Satureja bachtiarica Bunge.).

Components aRI-cal aRI-lit HDBP TSWD SDinnov HDRIFR MHD400 W MHD800 W MSHD400 W MSHD800 W HS-SPME ANOVA

Monoterpene hydrocarbons1 -Thujene 927 931 0.286 0.038b 0.250 0.046 0.187 0.051 0.223 0.011 0.170 0.014 0.095 0.007 0.143 0.030 0.130 0.036 p > 0.052 -Pinene 934 936 1.073 0.231 1.110 0.227 0.930 0.199 0.853 0.085 0.750 0.042 0.420 0.042 0.610 0.121 0.540 0.144 7.19 0.13 p < 0.013 Camphene 949 953 0.870 0.130 0.850 0.142 0.813 0.167 0.763 0.085 0.705 0.021 0.400 0.056 0.623 0.105 0.597 0.145 5.04 0.29 p < 0.014 -Pinene 979 980 0.133 0.023 0.130 0.020 0.107 0.023 0.120 0.010 0.110 0.000 0.060 0.000 0.090 0.010 0.093 0.021 0.85 0.03 p > 0.055 Myrcene 991 991 0.690 0.103 0.700 0.085 0.587 0.110 0.597 0.060 0.575 0.021 0.340 0.000 0.503 0.023 0.510 0.132 1.80 0.05 p < 0.016 -Phellandrene 1006 1005 0.177 0.029 0.187 0.050 0.130 0.026 0.133 0.015 0.120 0.000 0.080 0.000 0.113 0.006 0.113 0.029 0.50 0.00 p > 0.057 3-Carene 1012 1011 0.043 0.006 0.043 0.006 0.037 0.006 0.037 0.006 0.035 0.007 0.030 0.000 0.033 0.011 p > 0.058 -Terpinene 1018 1018 1.857 0.22b 1.757 0.24b 1.470 0.27bc 1.523 0.16bc 1.425 0.0bc 0.920 0.0d 1.290 0.043cd 1.290 0.27 cd 2.99 0.65a p < 0.019 -Cymene 1027 1026 14.043 1.75b 14.570 1.81b 13.880 2.06b 12.533 1.29b 13.975 0.11b 9.120 0.78c 12.806 0.38bc 12.887 2.61bc 32.7 3.98a p < 0.0510 Limonene 1029 1031 0.503 0.136 0.507 0.075 0.413 0.072 0.427 0.040 0.445 0.007 0.270 0.000 0.380 0.026 0.387 0.051 p > 0.0511 cis--Ocimene 1036 1040 0.133 0.055 0.113 0.030 0.077 0.015 0.077 0.015 0.070 0.000 0.045 0.007 0.070 0.010 0.080 0.017 p > 0.0512 trans--Ocimene 1047 1050 0.183 0.251 0.493 0.690 0.057 0.015 0.070 0.010 0.065 0.007 0.040 0.000 0.053 0.006 0.063 0.011 0.17 0.00 p < 0.0513 -Terpinene 1059 1062 12.653 1.18b 12.257 1.52b 10.163 1.8c 10.573 0.69bc 9.970 0.04c 7.210 0.38d 9.350 0.23c 9.747 1.41c 23.54 5.56a p < 0.0114 -Terpinolene 1089 1088 0.217 0.021 0.220 0.020 0.183 0.021 0.170 0.030 0.180 0.000 0.115 0.007 0.170 0.010 0.177 0.006 0.54 0.00 p < 0.0115 E,Z-Alloocimene 1131 1129 0.41 0.03 p < 0.01

32.861 4.17b 33.187 4.97b 29.034 4.83bc 28.099 2.49bc 28.595 0.27bc 19.115 1.278d 26.231 1cd 26.647 4.89cd 75.73 9.45a p < 0.01Oxygenated monoterpenesAlcohols16 Octan-3-ol 996 993 0.037 0.006 0.033 0.006 p < 0.0517 trans-Sabinene hydrate 1068 1068 0.067 0.021 0.237 0.038 0.247 0.060 0.203 0.075 0.250 0.000 0.160 0.000 0.303 0.045 0.310 0.010 1.16 0.43 p < 0.0118 Linalool 1101 1098 2.717 0.185b 3.05 0.151ab 3.040 0.451ab 3.080 0.170ab 2.820 0.014b 1.855 0.162c 3.377 0.319a 3.357 0.262a 5.49 1.11 p < 0.0519 Borneol 1167 1165 0.560 0.138 3.600 0.466 3.900 0.404 4.060 0.400 3.370 0.014 2.785 0.289 3.950 0.23 3.737 0.274 3.71 0.78 p < 0.0520 Terpinene-4-ol 1179 1177 0.560 0.020 0.447 0.049 0.420 0.046 0.540 0.132 0.365 0.035 0.275 0.007 0.430 0.026 0.427 0.061 p < 0.0521 -Cymene-8-ol 1187 1183 0.053 0.006 0.053 0.006 0.050 0.014 0.050 0.010 0.043 0.006 p > 0.0522 -Terpineol 1192 1189 0.130 0.017 0.127 0.025 0.140 0.010 0.147 0.023 0.150 0.042 0.100 0.000 0.127 0.006 0.113 0.032 p > 0.0523 cis-Piperitol 1181 1193 0.35 0.06 p < 0.0124 Geraniol 1257 1255 0.127 0.032 0.080 0.026 0.127 0.025 0.223 0.118 0.095 0.021 0.085 0.007 0.113 0.025 0.140 0.053 p > 0.05Phenol25 Thymol 1296 1290 28.610 2.99bc 26.806 2.07c 28.510 2.38bc 28.063 1.52bc 30.285 0.13b 34.525 1.48a 31.190 0.53b 30.990 1.88b 6.69 1.11d p < 0.0126 Carvacrol 1306 1298 24.980 1.07b 24.203 0.61b 24.827 2.75b 26.087 2.55b 26.885 0.95b 31.660 0.61a 27.300 0.52b 26.993 1.93b 4.80 0.45c p < 0.01Aldehydes27 trans-Chrysanthemal 1151 0.060 0.000 0.060 0.000 0.067 0.006 0.050 0.000 0.063 0.006 p < 0.05

814

S.M.

Mem

arzadeh et

al. /

Industrial Crops

and Products

76 (2015)

809816

Table 1 (Continued)

Components aRI-cal aRI-lit HDBP TSWD SDinnov HDRIFR MHD400 W MHD800 W MSHD400 W MSHD800 W HS-SPME ANOVA

28 Geranial 1276 1270 0.037 0.006 0.060 0.017 p < 0.05Ketone29 Camphor 1147 1143 0.047 0.006 0.043 0.006 p < 0.0530 Carvone 1246 1242 0.130 0.028 p < 0.05Esters31 Thymyl acetate 1355 1355 0.293 0.089 0.367 0.095 0.510 0.050 0.583 0.055 0.185 0.205 0.505 0.063 0.370 0.020 0.413 0.107 p > 0.0532 Carvacryl acetate 1374 1371 0.187 0.580 0.257 0.070 0.377 0.032 0.447 0.049 0.255 0.035 0.410 0.028 0.267 0.021 0.310 0.087 p > 0.05Ether33 Thymyl methyl ether 1244 1235 0.060 0.010 0.050 0.017 0.037 0.015 p > 0.0534 1,8-Cineole 1033 1033 0.08 0.030 0.073 0.023 0.160 0.084 0.067 0.006 0.063 0.011 p < 0.05Epoxides35 cis-Linalool oxide 1073 1074 0.033 0.006 p < 0.05

58.363 5.17b 59.307 3.63b 62.271 6.23b 63.547 5.1b 65.050 1.58b 72.360 2.64a 67.737 1.8c 67.059 4.75c 22.2 3.11d p < 0.01Sesquiterpene hydrocarbons36 -Caryophyllene 1422 1418 2.713 0.37b 3.870 0.51a 4.390 0.23a 3.023 0.14 b 2.355 0.19b 3.060 0.81b 2.250 0.02b 2.617 0.85b 2.08 0.22b p < 0.0137 Aromadendrene 1442 1439 0.097 0.015 0.147 0.025 0.167 0.006 0.075 0.007 0.105 0.035 0.073 0.006 0.093 0.032 p > 0.0538 Alloaromadendrene 1464 1461 0.077 0.011 0.087 0.032 p > 0.0539 -Humulene 1456 1452 0.243 0.071 0.340 0.026 0.203 0.011 0.155 0.007 0.230 0.042 0.150 0.000 0.220 0.091 p > 0.0540 -Bisabolene 1510 1509 0.160 0.036 0.253 0.060 0.310 0.035 0.200 0.026 0.130 0.014 0.210 0.070 0.120 0.000 0.157 0.073 p > 0.0541 cis--Bisabolene 1544 153642 -Bourbonene 1387 1384 0.065 0.049 p > 0.0543 -Gurjunene 1412 1409 0.043 0.006 0.073 0.011 0.080 0.000 0.047 0.006 0.040 0.000 0.050 0.014 0.030 0.000 0.043 0.015 p > 0.0544 Germacrene-d 1484 1480 0.065 0.049 p < 0.0545 -Cadinene 1526 1524 0.050 0.010 0.073 0.011 0.080 0.010 0.053 0.006 0.075 0.035 0.080 0.042 0.037 0.006 0.027 0.011 p > 0.05

3.063 0.44abc 4.659 0.68a 5.444 0.32a 3.613 0.22ab 2.960 0.35d 3.735 1.01bc 2.66 0.03cd 3.157 1.07cd 2.08 0.41d p < 0.01Oxygenated sesquiterpenesAlcohol46 (+) Spathulenol 1581 1576 0.083 0.035 0.083 0.038 0.160 0.030 0.223 0.144 0.090 0.000 0.290 0.113 0.107 0.035 0.120 0.070 p < 0.05Epoxide47 Caryophyllene oxide 1587 1581 0.467 0.150 0.590 0.219 1.090 0.166 1.283 0.710 0.595 0.007 1.595 0.445 0.713 0.175 0.810 0.410 p < 0.01

0.550 0.19c 0.673 0.26c 1.250 0.19abc 1.506 0.85 ab 0.685 0.01c 1.885 0.56a 0.820 0.21bc 0.93 0.48bc p < 0.05Other48 Tricyclene 923 926 0.033 0.006 0.033 0.006 0.037 0.006 0.030 0.000 0.030 0.00 0.023 0.006 p > 0.0549 Leden 1498 1490 0.137 0.011 0.143 0.006 0.127 0.055 0.095 0.035 0.105 0.035 0.063 0.006 0.080 0.036 p < 0.05Total 94.87 98 98.18 96.92 97.41 97.2 97.53 97.97 99.99

a RI (cal): retention indices determined on HP-5MS capillary column and RI (lit): retention indices of literature values.b Mean with different letter in a row are statistically signicant at 5% level probability), according to Duncans multiple range test. Values of major compounds are given as means SD.

S.M. Memarzadeh et al. / Industrial Crops and Products 76 (2015) 809816 815

Fig. 2. Compa thodsMeans with co est.

RIFR

than other mventional hand minimiMHD also ptillation suinvolve in acan be introoils.

Our resucould signitial oils fromfrom lossesof new comtion, and/orhighest conHS-SPME mof monoterassisted mehydrocarbohigh tempebe seen thaSDinnov, andmethods shrison of essential oil yield of savory (v/w on dry weight basis) by the extraction memmon letters are not signicant (p 0.05), according to Duncans multiple range t

HD7

8

9

10MSHD400 MHD400 MSHD800 MH D800

0

1

2

3

4

5

6

40 40 40 40 24 0

Ener

gy (K

wh)

Differe nt extracti on metho ds-ti m

Fig. 3. Energy consumption in different extract

ethods (Table 1). The MHD is more selective than con-ydrodistillation. Furthermore, the MHD is less tediouszes the risk of compound degradation due to heat. Theresents some advantages over conventional hydrodis-ch as extraction rate. Therefore, microwave does notny deterioration of the extracted components and itduced as a safe method for the extraction of essential

lts demonstrated that different extraction methodscantly change (p < 0.01) the chemical proles of essen-

the aerial parts of S. bachtiarica. These variations occur or increase in oil constituents due to the formationpounds by oxidation, glycoside hydrolysis, esterica-

other processer (Ghasemi-Pirbalouti et al., 2013b). Thetent of monoterpene hydrocarbons was obtained fromethod (75.73 9.45%), whereas the lowest percentagespene hydrocarbons were achieved from microwave-thods (Table 1). This results show that the monoterpenens had more susceptible than other chemical groups torature during the extraction process. In Table 1, it cant the essential oils obtained from HDBP, HDRIFR, TSWD,

MHD (400 W), MSHD (400 W), and MSHD (800 W)are similar compositions, except for the MHD (800 W)

and HS-SPM-cymene, ods. These dcompoundsnents, incluin agreemeet al., 2010of oxygenaplants (Roswere higheCompoundin various pounds likemoment wiextracted mhave low d(Bousbia et

3.3. Cost an

The reduproposed MHydrodistil.

HDBP SWDBP

SDinnov e240 240 240

e (min)

ion methods.

E methods. In MHD (800 W) method, the amounts of-terpinene, and linalool were lower than other meth-ifferences are probably due to the degradation of these

of monoterpene hydrocarbons into phenol compo-ding thymol and carvacrol (Table 1). This result wasnt with many other studies (Bousbia et al., 2009; Okoh) reported on other plant species, which the amountsted compounds in the essential oils of the aromaticmarinus ofcinalis L.) isolated by microwave-assistedr in comparison with conventional hydrodistillation.s with high and low dipolar moments could be extractedproportions by microwave extraction. Organic com-

oxygenated monoterpenes that have a high dipolarll interact more vigorously with microwaves and can beore easily in contrast with aromatic compounds whichipolar moments, such as monoterpene hydrocarbons

al., 2009).

d energy

ced cost of extraction is clearly advantageous for theHD and MSHD methods in terms of time and energy.lation and steam distillation using HDBP, HDRIFR, TSWD,

816 S.M. Memarzadeh et al. / Industrial Crops and Products 76 (2015) 809816

and SDinnov required an extraction time of 60 min for heating 1 L ofwater and 100 g of savory to the extraction temperature, followedby evaporation of water and essential oil for 180 min. The MAD andMSHD methods required heating for 5 min only and evaporation for25 min of the in situ water and essential oil of savory. The energyrequired to perform the different extraction methods shown inFig. 3. The power consumption was determined with a Wattmeterat the microwave generator entrance and the electrical heaterpower supply. In addition, microwave-assisted hydro-diffusion isa very clean method, which avoids residue generation and the useof a large quantity of water and voluminous extraction vessels.

4. Conclus

Extractioment of plaphenylpropextraction iable compoor large scawhich gave(MHD) met240 min fotion methoessential oiferent methand innovaistics analypower conextraction tIndeed, it g(240 min fotion methodconventionGenerally, emicrowaveenergy savicarvacrol), a

Acknowled

The authhis technicasupported bversity, Iran

References

Adams, R.P., 20Chromato

Bousbia, N., ViComparisohydrodisti355362.

Cronquist, A., York. Bota

Farhat, A., FabChemat, Ffrom oranChem. 125

Farhat, A., Ginies, C., Romdhane, M., Chemat, F., 2009. Eco-friendly and cleanerprocess for isolation of essential oil using microwave energy: experimentaland theoretical study. J. Chromatogr. A 1216, 50775085.

Ghasemi Pirbalouti, A., Nikobin Broujeni, V., Momeni, M., Malekpoor, F., Hamedi,B., 2011a. Antibacterial activity of Iranian medicinal plants againstStreptococcus iniae isolated from rainbow trout (Oncorhynchus mykiss). Arch.Biol. Sci. 63, 5966.

Ghasemi Pirbalouti, A., Hamedi, B., Abdizadeh, R., Malekpoor, F., 2011b. Antifungalactivity of the essential oil of Iranian medicinal plants. J. Med. Plant. Res. 5,50895093.

Ghasemi Pirbalouti, A., Pirali, E., Pishkar, G.H., Jalali, S.M.A., Reyesi, M., JafarianDehkordi, M., Hamedi, B., 2011c. The essential oils of some medicinal plants onthe immune system and growth of rainbow trout (Oncorhynchus mykiss). J.Herb. Drugs 2, 149155.

Ghasemi Pirbalouti, A., Dadfar, S., 2013. Chemical constituents and antibacterialactivity of essential oil of Satureja bachtiarica (Lamiaceae). Acta Pol. Pharm. 70,

938.i Pirbacts of dtiari si Pirbaitative

Chemi Pirbapositi282i Pirbaical

s agai207i Pirbaity, tots useani, Modistiactionn, B.,ucts: ochemM., 20

Saturrimeni, M.E.owav

techneini, Ffferenacteri10.108rian, Aish F, pp. 5.O., Sabacterydrod. 120

ssartiential osoundmato, F., Jaes. 12ko, E.Eaction

(Mill.oxidanKordjaseful t147.A., Fergravit7..F., Ya

abolite. 22, 6ion

n is one of the crucial steps for research and develop-nt secondary metabolites, such as avonoids, quinones,anoids, terpenoids and alkaloids. Microwave assisteds one of the important techniques for extracting valu-unds from herbs, and it is quite adaptable on a smallle (i.e., on a laboratory or industry scale). The method

the best results was the microwave hydro-diffusionhod which gave reduced extraction time (30 min againstr conventional hydro-distillation and steam distilla-ds) and gave no differences in essential oil yield. Thels from savory have been extracted using seven dif-ods and a comparison of the different conventionaltive techniques has been carried out. The character-zed were extraction time, essential oil composition,sumption and yield. Results identied the optimalechnique as being microwave hydro-diffusion (MHD).ave the maximum yield (1.4% v/w) in only 30 minr conventional hydro-distillation and steam distilla-s) and consumed 0.30.5 kWh (against 6.68.3 kWh for

al hydro-distillation and steam distillation methods).xtraction of the essential oil from S. bachtiarica with

-assisted hydro-diffusion (MHD) was better in terms ofng, extraction time, oxygenated fraction (thymol andnd product quality.

gments

ors would like to acknowledge Mr. Behzad Hamedi forl assistance in essential oil extraction. This work wasy Deputy of Research & Technology, Islamic Azad Uni-.

07. Identication of Essential Oil Components by Gasgraphy/Mass Spectrometry, 4th Ed. Allured Publishing Corporation.an, M.A., Ferhat, M.A., Petitcolas, E., Meklati, B.Y., Chemat, F., 2009.n of two isolation methods for essential oil from rosemary leaves:llation and microwave hydrodiffusion and gravity. Food Chem. 114,

1988. The Evolution and Classication of Flowering Plants, The Newnical Garden, New York, USA.iano-Tixier, A.S., El Maataoui, M., Maingonnat, J.F., Romdhane, M.,., 2011. Microwave steam diffusion for extraction of essential oilge peel: kinetic data, extracts global yield and mechanism. Food, 255261.

933Ghasem

EffeBakh

GhasemqualFood

Ghasemcom26, 2

GhasemChemherb2063

Ghasemactivplan

Golmakhydrextr

KaufmanprodPhyt

Khajeh, fromexpe

Lucchesmicrnew

Mirhossof dicharorg/

MozaffaEnglIran

Okoh, Oantiby hChem

Prino-IesseultraChro

SedkonOil R

Stashenextralbaanti

Tigrine-an u141

Vian, M.and 141

Zhang, HmetTechlouti, A., Oraie, M., Pouriamehrc, M., Solaymani Babadi, E., 2013a.rying methods on qualitative and quantitative of the essential oil ofavory (Satureja bachtiarica Bunge.). Ind. Crops Prod. 46, 324327.louti, A., Mahdad, E., Craker, L., 2013b. Effects of drying methods on

and quantitative properties of essential oil of two basil landraces.. 141, 24402449.louti, A., Vosoghi, N., Craker, L., Shirmardi, H.A., 2014a. Chemicalon of the essential oil of Satureja kallarica Jamzad. J. Essent. Oil Res.31.louti, A., Neshat, S.H., Rahimi, E., Hamedi, B., Malekpoor, F., 2014b.composition and antibacterial activity of essential oils of Iraniannst Staphylococcus aureus isolated from milk. Int. J. Food Prop. 17,1.louti, A., Siahpoosh, A., Setayesh, M., Craker, L., 2014c. Antioxidanttal phenolic and avonoid contents of some medicinal and aromaticd as herbal teas and condiments in Iran. J. Med. Food 17, 11511157..T., Rezaei, K., 2008. Comparison of microwave-assistedllation with the traditional hydrodistillation method in the

of essential oils from Thymus vulgaris L. Food Chem. 109, 925930. Christen, P., 2002. Recent extraction techniques for naturalmicrowave-assisted extraction and pressurised solvent extraction.. Anal. 13, 105113.

11. Optimization of process variables for essential oil componentseja hortensis by supercritical uid extraction using Box-Behnkental design. J. Supercrit. Fluids 55, 944948., Smadja, J., Bradshaw, S., Louw, W., Chemat, F., 2007. Solvent freee extraction of Elletaria cardamomum L.: a multivariate study of aique for the extraction of essential oil. J. Food Eng. 79, 10791086.., Rahimmalek, M., Ghasemi Pirbalouti, A., Taghipoor, M., 2015. Effectt drying treatments on essential oil yield, composition and colorstics of Kelussia odoratissima Mozaff. J. Essent. Oil Res., http://dx.doi.0/10412905.2015.1015691., 2008. A Pictorial Dictionary of Botany Botanical Taxonomy Latin rench Germany Persian/Complied. Farahang Moaser, Tehran,22.dimenko, A.P., Afolayan, A.J., 2010. Comparative evaluation of theial activities of the essential oils of Rosmarinus ofcinalis L. obtainedistillation and solvent free microwave extraction methods. Food, 308312.r, S., Ginies, C., Cravotto, G., Chemat, F., 2013. A comparison ofils obtained from Lavandin via different extraction processes:, microwave, turbo hydrodistillation, steam and hydrodistillation. J.

gr. A 1305, 4147.mzad, Z., 2000. Essential oil of Satureja bachtiarica Bunge. J. Essent., 545546.., Jaramillo, B.E., Martnez, J.R., 2004. Comparison of different

methods for the analysis of volatile secondary metabolites of Lippia) NE Brown, grown in Colombia, and evaluation of its in vitrot activity. J. Chromatogr. A 1, 93103.ni, N., Meklati, B.Y., Chemat, F., 2006. Microwave dry distillation asool for extraction of edible essential oils. Int. J. Aromather. 16,

nandez, X., Visinoni, F., Chemat, F., 2008. Microwave hydrodiffusiony, a new technique for extraction of essential oils. J. Chromatogr. A,

ng, X.H., Wang, Y., 2011. Microwave assisted extraction of secondarys from plants: current status and future directions. Trends Food Sci.72688.

Chemical composition and yield of essential oils from Bakhtiari savory (Satureja bachtiarica Bunge.) under different extra...1 Introduction2 Material and methods2.1 Plant material2.2 Chemicals2.3 Extraction procedures2.3.1 Hydrodistillation using two Clevenger apparatus2.3.2 Traditional steam and water distillation procedure2.3.3 Steam distillation apparatus and procedure2.3.4 Microwave-assisted steam hydro-diffusion (MSHD) apparatus and procedure2.3.5 Microwave-assisted hydro-diffusion (MHD)2.3.6 Head space solid-phase microextraction (HS-SPME)

2.4 Identification of the oil constituents2.5 Statistical analyses

3 Results and discussion3.1 Effect of extraction methods on the essential oil yield3.2 Effect of extraction methods on the essential oil composition3.3 Cost and energy

4 ConclusionAcknowledgmentsReferences