research article chemical composition and...

Research ArticleChemical Composition and Antioxidant and AntimicrobialActivities of Wormwood (Artemisia absinthium L) EssentialOils and Phenolics

Kamel Msaada12 Nidhal Salem1 Olfa Bachrouch1 Slim Bousselmi1

Sonia Tammar1 Abdulkhaleg Alfaify2 Khaldoun Al Sane2 Wided Ben Ammar3

Sana Azeiz1 Adel Haj Brahim1 Majdi Hammami1 Sawsen Selmi1

Ferid Limam1 and Brahim Marzouk1

1Laboratory of Bioactive Substances Biotechnology Center in Borj-Cedria Technopol BP 901 2050 Hammam-Lif Tunisia2Biology Department College of Sciences King Khalid University PO Box 9004 Abha 61413 Saudi Arabia3Unit of Research ldquoNitrogenized Nutrition and Metabolism and Stress Proteinsrdquo UR 9909-20 Department of BiologyFaculty of Science University of Tunis 2092 Tunis Tunisia

Correspondence should be addressed to Kamel Msaada msaada kamelyahoofr

Received 15 December 2014 Revised 15 February 2015 Accepted 16 February 2015

Academic Editor Augusto C Tome

Copyright copy 2015 Kamel Msaada et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

The aimof this studywas to determine the chemical variability of wormwood extracts as affected by the growing region Antioxidantand antimicrobial activities were also investigated The essential oil composition variability of A absinthium L aerial partscollected from four different Tunisian regions was assessed by gas chromatography (GCFID) and by gas chromatography massspectrometry (GCMS) In addition total polyphenols flavonoids and condensed tannins as well as antioxidant antibacterial andantifungal activities ofmethanolic extract and essential oils were undertaken Chromatographic analysis of wormwood essential oilsshowed the predominance of monoterpene hydrocarbons represented mainly by chamazulene RP-HPLC analysis of wormwoodmethanolic extract revealed the predominance of phenolic acids Antiradical activity was region-dependant and the methanolicextract of Bou Salem region has the strongest activity (CI

50= 938 plusmn 082 120583gmL) Concerning the reducing power the methanolic

extract of Bou Salem Jerissa and Boukornine regions was more active than the positive control Obtained results of antimicrobialactivities showed that wormwood essential oil is endowed with important antibacterial activity which was strongly related to theorganoleptic quality of oil which appeared strongly region-dependant A absinthium L EOs investigated are quite interesting froma pharmaceutical standpoint because of their biological activities

1 Introduction

Artemisia absinthium L (Asteraceae) commonly known aswormwood in United Kingdom and absinthe in France is anaromatic perennial small shrub It is locally known as ldquochajretmariemrdquo in Tunisia The herb has always been of a greatbotanical and pharmaceutical interest and is employed infolkmedicine against various paints [1]Wormwood essentialoil has been widely used mainly for its neuroprotective [2]antifungal [3] antimicrobial [4] insecticidal [3] acaricidal[5] anthelmintic [6] antimalarial [7] hepatoprotective [8]

and antidepressant [9] proprieties In addition the herb isused tomake a tea for helping pregnantwomenduring pain oflabor and in treating leukaemia and sclerosis [10] Moreoverthe organic extract of this herb revealed toxic and antifeedanteffect on Leptinotarsa decemlineata [11] It was always usedas a drink in France called absinthe which caused dementiaIn the 18th century alcoholic decoctions of wormwood andother plants were used as all-purpose remedies or ldquocure-allsrdquobut it was not until the beginning of the 19th century that thewormwood-flavored alcoholic extracts and distillates wereseen not only as potent medicines but also as aperitifs and

Hindawi Publishing CorporationJournal of ChemistryVolume 2015 Article ID 804658 12 pageshttpdxdoiorg1011552015804658

2 Journal of Chemistry

the large scale production of absinthe began In additionArtemisia absinthium L was used freshly for the plant speciesas well as for the alcoholic beverage

Aromatic and medicinal plants are an easily accessibleand edible source of natural antioxidants They have theability to protect the organism from damage caused byfree radical-induced oxidative stress such as cancer andcardiovascular and neurodegenerative diseases [12 13]

Various studies have reported previously that reactiveoxygen species (ROS) are highly reactive and cause damageto protein lipids enzymes and DNA [14] In this contextthe use of plants and herbs in food pharmaceutical andcosmetic industries as a source of natural antioxidant andbiologically active compounds [15 16] has attracted a greatdeal of scientific interest In addition Pietta [17] reported thatthe antioxidant effect of plant products is mainly attributed tophenolic compounds

In the food industry a large spectrum of microorgan-ism leads to food spoilage Therefore preservation of foodmaterial from degradation mainly by oxidation processesand microorganism activity during production storage andmarketing is an important stage Also Friedman et al[18] reported the increase of some pathogensrsquo resistance tosynthetic antibiotics which are also uncomfortable to patientsdue to their adverse drug reactions

That is why the use of plants and herbs as a sourceof natural products has attracted the interest of manyresearchers They have been screened for their potential usesas alternative remedies for the treatment of many infectionsas an antioxidant and also as a natural food preservative[19 20]

Many authors have reported composition and antioxidantand antibacterial proprieties of A absinthium L essentialoil and extracts Wright [21] has reported that wormwoodstimulant propriety is caused by bitter substances as artabsin(sesquiterpene lactone) and absinthin (dimer of sesquiter-pene lactone) present in plant extracts In fact Iranian worm-wood essential oil was characterized by the predominance of120573-pinene and 120573-thujone [22] In the same context essentialoil of A absinthium L collected from Morocco is essentiallyrepresented mainly by 120572-thujone (3969) sabinyl acetate(1096) and 120573-thujone (725) [23] Furthermore Martınet al [24] showed that the major compounds of wormwoodfound in the SFE extracts as well as in the hydrodistilledessential oils were Z-epoxyocimene chrysanthenol andchrysanthenyl acetate

To the best of our knowledge there are no reportsdealing with chemotype A absinthium L essential oil inTunisia In our investigation we have studied essential oiland methanolic extracts composition of wild populationof wormwood cultivated in Tunisia and harvested fromfour distinct regions (Jerissa Bou Salem Boukornine andKairouan) Furthermore we have evaluated their antioxidantantibacterial and antifungal activities The purpose of thisstudy was to underline the variability of wormwood essentialoil and methanolic extracts and their biological activities asaffected by the collection site

2 Material and Methods

21 Chemicals All solvents used in the experiments (diethylether acetonitrile and water of high-performance liquidchromatography (HPLC) grade and ethanol methanoland ethyl acetate of analytical grade) were purchasedfrom Merck (Darmstadt Germany) Sodium hydroxide(NaOH) sodium phosphate (Na

2HPO4) sodiummonobasic

phosphate anhydrous (NaH2PO4H2O) sodium carbonate

(Na2CO3) sodium nitrite NaNO

2 butylated hydroxytoluene

(BHT) 120573-carotene linoleic acid ethylenediaminetetraaceticacid (EDTA) 3-(2-pyridyl)-56-bis(4-phenyl-sulphonicacid)-124-triazine (ferrozine) iron(II) chloride tetrahydrate(FeCl2sdot4H2O) iron(II) chloride (FeCl

2) iron(III) chloride

(FeCl3) 11-diphenyl-2-picrylhydrazyl (DPPH) polyvinyl-

polypyrrolidone Folin-Ciocalteu reagent potassium ferri-cyanide (K

3Fe(CN)

6) aluminium chloride (AlCl

3) homolo-

gous series of C6ndashC17

n-alkanes and high-purity standardsof essential oil were purchased from Sigma-Aldrich(Steinheim Germany) Hydrochloric acid (HCl) (36ndash38) and potassium hydroxide (KOH) were provided by JT Baker (Deventer Netherlands) Trifluoroacetic acid (TFA)(999) was purchased from ROMIL Ltd (Cambridge UK)Cyanidin 3-O-glucoside was purchased from Extrasynthese(Genay France) Deionized water was used to prepare allsolutions unless otherwise indicated These solutions werewrapped in aluminium foil and stored at 4∘C All otherchemicals used were of analytical grade

22 Plant Material A absinthium L aerial parts were har-vested randomly at flowering stage from different Tunisianregions The choice of these sites was dictated by the geo-graphical differences of the species studied (Table 1) Theplant material was harvested from four regions (Jerissa BouSalem Boukornine andKairouan)Theharvested aerial partswere identified by Professor Abderrazek Smaoui (Borj CedriaBiotechnology Center Tunisia) according to the Tunisianflora and a voucher specimen was deposited at the herbariumof the Laboratory of Bioactive Substances (BiotechnologyCenter of Borj Cedria) under the number Aab21213 Theharvested material was freeze-dried and then ground to finepowder by an electric mill and conserved in a dessicator atroom temperature (sim25∘C) in darkness for further uses

23 Essential Oil Extraction The leaves stems and flowerswere cut into small pieces and subjected to hydrodistillationtype Clevenger for 90min in accordance with EuropeanPharmacopoeia method [25] Essential oil extractions weredone in triplicate for each A absinthium L collection siteYield percentagewas calculated asmg of essential oil per 100 gof plant dry matter All experiments were done in triplicate

24 Gas Chromatography (GC) Analysis Analysis of Aabsinthium L essential oil volatile compounds by gas chro-matography (GC) was carried out on a Hewlett-Packard6890 gas chromatograph (Palo Alto CA USA) equippedwith a flame ionization detector (FID) and an electronic

Journal of Chemistry 3

Table 1 Geographical and bioclimatic collection sites parameters

Longitude Latitude Elevation (m) Bioclimatic stageBoukornine 9∘471015840564110158401015840E 36∘421015840253310158401015840N 70 Superior semiaridJerissa 8∘341015840521210158401015840E 35∘541015840383410158401015840N 647 Superior semiaridBou Salem 8∘59101584023910158401015840E 36∘38101584085710158401015840N 247 SubhumidKairouan 9∘471015840564110158401015840E 35∘371015840408610158401015840N 191 Superior aridE east N north

pressure control (EPC) injector A polar polyethylene gly-col (PEG) HP-INNOWax and a 5 diphenyl and 95dimethylpolysiloxane apolar HP-5 capillary columns (30m times025mm 025mm film thickness Hewlett-Packard CAUSA) were used The flow of the carrier gas (N

2) was

16mLmin The split ratio was 60 1 The analysis wasperformed using the following temperature program oventemperature kept isothermally at 35∘C for 10min increasedfrom 35 to 205∘C at the rate of 3∘Cmin and kept isothermallyat 205∘C for 10min Injector and detector temperatures wereheld at 250 and 300∘C respectively The individual peakswere identified by retention times and retention indices(relative to C

6ndashC22

n-alkanes) compared with those ofknown compounds Percentage composition of essential oilswas computed from GC-FID peak areas without correctionfactor

25 Gas Chromatography-Mass Spectrometry (GCMS) Anal-ysis Volatile compounds analysis by GCMS was performedon a gas chromatograph HP 5890 (II) interfaced with a HP5972 mass spectrometer (Palo Alto CA USA) with electronimpact ionization (70 eV) A HP-5 MS capillary column(30m times 025mm coated with 5 phenyl methyl silicone and95 dimethylpolysiloxane 025mm film thickness Hewlett-Packard CA USA) was used The column temperature wasprogrammed to rise from 50 to 240∘C at a rate of 5∘CminThe carrier gas was helium with a flow rate of 12mLminsplit ratio was 60 1 Scan time and mass range were 1 s and40ndash300mz respectively

26 Preparation of Plant Extracts Sample extracts wereobtained by stirring 1 g of dry aerial parts (leaves stems andflowers) powder with 10mL of methanol according to themethod of Mau et al [26] Obtained extracts were kept for24 h at +4∘C filtered through a Whatman number 4 filterpaper and freed of solvent under reduced pressure at 45∘Cusing a rotary evaporator and then lyophilized The driedcrude concentrated extracts were stored at minus20∘C until usedfor analyses

27 Total Phenolic Content The total phenolic content wasassayed using the Folin-Ciocalteu reagent following Sin-gletonrsquos method slightly modified by Dewanto et al [27]An aliquot (0125mL) of a suitable diluted methanol samplewas added to 05mL of deionized water and 0125mL ofthe Folin-Ciocalteu reagent The mixture was shaken andallowed to stand for 6min before adding 125mL of 7sodium carbonate (Na

2CO3) solution The solution was then

adjusted with deionized water to a final volume of 3mL andmixed thoroughly After incubation for 90min at 23∘C theabsorbance versus prepared blank was read at 760 nm Totalphenolic contents of leaves stems andflower (three replicatesper treatment) were expressed as milligrams of gallic acidequivalents per gram of dry weight (mg of GAEg of DW)through the calibration curve with gallic acidThe calibrationcurve range was 50ndash400mgmL (1198772 = 099) Triplicatemeasurements were taken for all samples

28 Total Condensed Tannins Content The total tannincontent was measured using the modified vanillin assaydescribed by Sun et al [28] A total of 3mL of 4 methanolvanillin solution and 15mL of concentrated H

2SO4were

added to 50 120583L of suitably diluted sample The mixture waskept for 15min and the absorbance was measured at 500 nmagainst methanol as a blank The amount of total condensedtannins was expressed as milligrams of (+)-catechin equiv-alent per gram of dry weight (mg of CEg of DW) throughthe calibration curve with catechin Triplicate measurementswere taken for all samples

29 Total Flavonoid Content The total flavonoid content wasmeasured according to Dewanto et al [27] A total of 250120583Lof the sample appropriately diluted was mixed with 75120583Lof 5 NaNO

2(sodium nitrite) After 6min 150 120583L of 10

aluminum chloride (AlCl3) and 500 120583L of 1M NaOH were

added to the mixture Finally the mixture was adjusted to25mL with distilled water The absorbance versus preparedblank was read at 510 nm Total flavonoid contents of aerialparts (three replicates per treatment) were expressed asmilligrams of catechin equivalents per gram of dry weight(mg of CEg of DW) through the calibration curve withcatechinThe calibration curve range was 50ndash188 500mgmLTriplicate measurements were taken for all samples

210 Reversed-Phase- (RP-) HPLC Evaluation of MajorAntioxidant Compounds Phenolic compounds analysis wascarried out using an Agilent Technologies 1100 series liquidchromatograph (RP-HPLC) coupled with an UV-Vis multi-wavelength detector The separation was carried out on 250times 46mm 4120583m Hypersil ODS C

18reversed phase column

The mobile phase consisted of acetonitrile (solvent A) andwater with 02 sulphuric acid (solvent B) The flow rate waskept at 05mLmin The gradient programme was as follows15A85B 0ndash12min 40A60B 12ndash14min 60A40B 14ndash18min 80 A20 B 18ndash20min 90 A10 B 20ndash24min and 100 A 24ndash28min The injected volume was


20120583L and peaks were monitored at 280 nm Peaks wereidentified by congruent retention times compared with thoseof authentic standards

211 Antioxidant Activities

2111 DPPHAssay Theelectron donation ability of the aerialparts extracts was measured by bleaching of the purple-coloured solution of 11-diphenyl-2-picrylhydrazyl radical(DPPH) according to the method of Hatano [29] One-halfmL of 02mMDPPHmethanolic solutionwas added to aerialparts extracts of A absinthium L (2mL 10ndash1000 120583gmL)After an incubation period of 30min at room temperaturethe absorbance was read against a blank at 517 nm The inhi-bition percentage of free radical DPPH (IP) was calculatedas follows

IP = [(119860blank minus 119860 sample)

119860blank] times 100 (1)

where 119860blank is the absorbance of the control reaction and119860 sample is the absorbance in the presence of plant extractExtract concentration providing 50 inhibition (IC

50) was

calculated from the regression equation prepared from theconcentration of the extracts and the inhibition percentageBHT was used as a positive control

2112 Reducing Power Themethod of Oyaizu [30] was usedto assess the reducing power of aerial parts extracts of Aabsinthium L These extracts (1mL) were mixed with 25mLof a 02M sodium phosphate buffer (pH = 66) and 25mLof 1 potassium ferricyanide (K

3Fe (CN)

6) and incubated

in a water bath at 50∘C for 20min Then 25mL of 10trichloroacetic acid was added to the mixture that wascentrifuged at 650 g for 10minThe supernatant (25mL) wasthen mixed with 25mL distilled water and 05mL of 01ferric chloride solution The intensity of the blue-green colorwas measured at 700 nmThe extract concentration at whichthe absorbance was 05 for the reducing power (EC

50) was

obtained from the linear regression equation prepared fromthe concentrations of the extracts and the absorbance valuesHigh absorbance indicates high reducing power Ascorbicacid was used as a positive control

212 Screening of Antibacterial and Antifungal ActivitiesAntibacterial activity was analyzed by the disc diffusionmethod [31] against three human pathogenic bacteria includ-ing Staphylococcus aureus 25923 Staphylococcus methicillin-resistant andListeriamonocytogenes All bacteriawere grownon LB plate at 30∘C for 18ndash24 h previous inoculation onto thenutrient agar A loop of bacteria from the agar slant stockwas cultivated in nutrient broth overnight and spread with asterile cotton swap onto Petri dishes containing 10mL of APIsuspension medium and adjusted to the 05 McFarland tur-bidity standards with a Densimat (bioMerieux) Sterile filterpaper discs (6mm in diameter) impregnated with 20 120583L ofplant extract (10mgmL) were placed on the cultured platesAfter 1-2 h at 4∘C the treated Petri dishes were incubated at

37∘C for 18ndash24 h The solvents acetonewater 2 8 and puremethanol without extracts served as negative controls andtetracycline was used as the positive one The antimicrobialactivity was evaluated by measuring the diameter of thegrowth inhibition zone around the discs Each experimentwas carried out in triplicate and the mean diameter of theinhibition zone was recorded

The same agar-disc diffusionmethodwas used for screen-ing the antifungal activity of A absinthium L aerial partsextracts Five yeast strains (Fusarium graminearum Fusariumculmorum Fusarium oxysporum Sclerotinia and Rhizoctoniasolani) were first grown on Sabouraud chloramphenicol agarplate at 30∘C for 18ndash24 h Several colonies of similarmorphol-ogy of the clinical yeast were transferred into API suspensionmedium and adjusted to 2McFarland turbidity standard witha Densimat (bioMerieux) The inocula of the respective yeastwere streaked on to Sabouraud chloramphenicol agar platesat 30∘C using a sterile swab and then dried A sterilized 6mmpaper disc was loaded with 20120583L (10mgmL) of aerial partsextract The treated Petri dishes were placed at 4∘C for 1-2 hand then incubated at 37∘C for 18ndash24 h The inhibition offungal growth was also evaluated by measuring the diameterof the transparent inhibition zone around each disc Percentinhibition (119868) was calculated as 119868 = [(119862 minus 119879)119862] times100 where 119862 is the diameter of the control colonies and119879 is the diameter of the test colonies The average of threemeasurements was taken The susceptibility of the standardwas determined using a disc paper containing Nystatin

213 Data Analysis All analyses were performed in triplicateand the results are expressed as mean values plusmn standarddeviations (SD)The data were subjected to statistical analysisusing statistical program package STATISTICA [32] Theone-way analysis of variance (ANOVA) followed by Duncanmultiple range test was employed and the differences betweenindividual mean values were deemed to be significant at 119875 lt005 In addition a principal component analysis (PCA) wasperformed in order to discriminate between different regionon the basis of their essential oils and phenolic composition

3 Results and Discussion

31 Essential Oil Yield The hydrodistillation extraction of100 g of wormwood dry aerial parts collected from fourdifferent regions yielded a dark blue essential oilThese yieldswere illustrated in Figure 1 The results show that EO yieldvaries significantly (119875 lt 005) according to region factorThereby the optimal yield was observed at the region of BouSalem (146 plusmn 006) followed by the region of Kairouan(112 plusmn 008) and Boukornine and Jerissa regions (110 plusmn004 and 100 plusmn 003 resp) Similar results were obtainedby Orav et al [33] at different European regions and theoil yields were ranged from 01 to 11ww In addition theEO yields detected in the present study were higher thanthose determined by Lopes-Lutz et al [34] in the regionof the central prairies of Alberta (Canada West) that theyfound (05 ww) which justifies the effect of region on theEO yield The impact of other environmental factors on EO


a

bc

bc

0

02

04

06

08

1

12

14

16

18

JeacuterissaBoukornineKairouanBou Salem

Esse

ntia

l oil

yiel

d (

ww

)

Figure 1 Essential oil yield of the aerial parts of A absinthiumL collected from four Tunisian regions Values with differentsuperscripts (andashc) are significantly different at 119875 lt 005

yield was also described by Msaada et al [35] and Msaadaet al [36 37] In addition seasonal and regional variationswere observed in the EO yield of coriander [38] The sameauthors also reported the effect of extraction techniques onthe essential oils yield and composition [39]

32 Variability of Chemical Composition of EO Thehydrodistillation of wormwood aerial parts collected fromfour different Tunisian regions provides an EO with blue-black color This latter was due to the predominance ofchamazulene component which gives the oil with a highorganoleptic quality [33] The results of chromatographicanalysis by gas chromatography (GCFID) and by couplinggas chromatography to mass spectrometry (GCMS) arepresented in Table 2

Essential oils analyzed are divided into eight classes basedon their chemical functional groups A total of 56 compoundswere identified representing 9872 plusmn 1023 9764 plusmn 9789855 plusmn 988 and 9883 plusmn 976 of total volatiles in theregions of Jerissa Boukornine Kairouan and Bou Salemrespectively These different identified compounds vary sig-nificantly (119875 lt 005) from region to another and are highly(119875 lt 0001) affected by the regional factor (Table 2)

The major contribution was attributed to the monoter-pene hydrocarbons fraction which represents 5602 plusmn6214531 plusmn 531 4573 plusmn 522 and 6208 plusmn 749 of all com-pounds detected in Jerissa Boukornine Kairouan and BouSalem regions respectively The latter fraction is dominatedby chamazulene where its maximum rate (3993 plusmn 456)was observed in the Jerissa region followed by the regionsof Bou Salem (3481 plusmn 411) Kairouan (2951 plusmn 326) andBoukornine (2530plusmn312)These results were in agreementwith the literature However Kordali et al [40] showed thatessential oils chemical composition of wormwood collectedin Turkey was dominated by chamazulene (178) followedby nuciferol butanoate (82) nuciferol propionate (51)and caryophyllene oxide (43) In this work 120573-thujonea monoterpene ketone was detected with significant per-centages 2209 plusmn 212 in the region of Kairouan up to

672 plusmn 075 in the region of Bou Salem Meanwhile thechemical composition of essential oils of wormwood growingin the USA shows that the 120573-thujone (175 to 423) and cis-sabinyl acetate (151 to 534) are the major compounds [41]In agreement with our results previous studies have shownthat the rate of thujone in essential oils of wormwood isgreater than 35 [42] In addition in the wormwood samplescollected from different European countries (Greece SpainUkraine France and Italy) 120572-thujone and 120573-thujone weredetected asmajor componentswith percentages ranging from45 to 387 [33]

It should be noted that the sesquiterpene fractionwhich is represented by 120572-humulene the 120574-muurolenethe 120574-curcumene the 120572-copaene the 120573-selinene the 120572-bisabolene the 120574-cadinene the germacrene D the 120572-calacorene caryophyllene oxide and 120572-bisabolol rangesfrom 8 to 14 of total volatiles This slight variation could beexplained by similar environmental and genetic factors Anearlier study showed that the rate of sesquiterpenes detectedwas 22 [34] rate lower than that found in this workMonoterpene alcohols (linalool lavandulol terpinen-4-ol 120572-terpineol nerol sabinol trans-p-cymene-8-ol eugenol bor-neol and geraniol) were highly affected by the region factor(Table 2)

Principal component analysis was carried out in orderto determine the relationship between the different regionsof plant collection on the basis of their essential oil compo-sition A better discrimination was revealed on the three-dimensional visualization of the plotted scores Resultsobtained from the PCA (Figure 2) showed the existence ofone well-defined group represented by the regions JerissaBoukornine and Kairouan suggesting similar compositionsThe Bou Salem region was clearly distinguished from thelatter group both in quality and in quantity

In conclusion the Artemisia absinthium L plants col-lected from four Tunisian habitats biosynthesized essentialoils of one chemotype chamazulene

33 Total Polyphenols Flavonoids and Condensed TanninsContents Total phenol assay is used as a routing assay toassess phenolic antioxidants due to its simplicity and repro-ducibility Phenolic compounds have the ability to reduceFolin-Ciocalteu reagent and the reaction may be followed bymeasuring a change in the color of the solution from intenseyellow to blue spectrophotometrically Quantitative and qual-itative changes of the antioxidant compounds with functionto region effect hold great significance from both dietary andnutritional points of view while phenolic compounds areused as alternatives to synthetic drugs and were shown to besafe for use in processed foods [43]

Phenolic quantification revealed a wide range of aerialpartsrsquo polyphenol contents as function of the region as shownin Figure 3The total polyphenol contents varied significantly(119875 le 005) from one region to another and reached themaximum in the region of Kairouan (9989plusmn330mgGAEgDW) followed by the region of Bou Salem Boukornine andJerissa where their levels were 8370 plusmn 131 7205 plusmn 183 and4939 plusmn 220mg GAEg DM respectively Monica et al [44]


Table 2 ANOVA analysis qualitative and quantitative essential oil composition (peak area plusmn SD) of A absinthium L aerial parts

Compoundlowast RRIa RRIb Collection region df 119865 119875

Jerissa Boukornine Kairouan Bou SalemTricyclene 1014 927 100 plusmn 011a 002 plusmn 000d 074 plusmn 006b 023 plusmn 003c 3 96733 0004lowastlowastlowast

120572-Thujene 1032 939 005 plusmn 001c 005 plusmn 000c 328 plusmn 032a 023 plusmn 002b 3 1007777 0000lowastlowastlowast

120572-Pinene 1035 931 010 plusmn 001c 002 plusmn 000b 062 plusmn 005a 020 plusmn 003b 3 6581 0000lowastlowastlowast

Camphene 1076 950 006 plusmn 001c 003 plusmn 000d 066 plusmn 007a 024 plusmn 003b 3 16301 0000lowastlowastlowast

120573-Pinene 1118 980 120 plusmn 010a 025 plusmn 002c 002 plusmn 000d 027 plusmn 003b 3 25196 0000lowastlowastlowast

Sabinene 1132 976 028 plusmn 003c 187 plusmn 019a 002 plusmn 000d 167 plusmn 015b 3 338383 0000lowastlowastlowast

Myrcene 1176 988 050 plusmn 004b 005 plusmn 001c 152 plusmn 014a 002 plusmn 000d 3 55519 0000lowastlowastlowast

120572-Terpinene 1188 1018 003 plusmn 000c 044 plusmn 005a 008 plusmn 001b 003 plusmn 000c 3 24778 0000lowastlowastlowast

Limonene 1203 1026 010 plusmn 001d 095 plusmn 008a 014 plusmn 001c 040 plusmn 003b 3 3949 0000lowastlowastlowast

18-Cineole 1213 1033 003 plusmn 000c 003 plusmn 000c 035 plusmn 004a 008 plusmn 001b 3 8314 0000lowastlowastlowast

120573-Phellandrene 1218 1031 010 plusmn 002b 004 plusmn 000d 006 plusmn 001c 026 plusmn 003a 3 2544 0000lowastlowastlowast

Z-120573-Ocimene 1246 1040 020 plusmn 003c 159 plusmn 016a 026 plusmn 003b 005 plusmn 000d 3 53884 0000lowastlowastlowast

E-120573-Ocimene 1266 1050 008 plusmn 001d 010 plusmn 002c 140 plusmn 016a 043 plusmn 005b 3 10586 0000lowastlowastlowast

120574-Terpinene 1267 1062 010 plusmn 002b 007 plusmn 001c 018 plusmn 002a 004 plusmn 000d 3 2718 0000lowastlowastlowast

p-Cymene 1280 1026 008 plusmn 001c 012 plusmn 001b 022 plusmn 003a 004 plusmn 000d 3 4475 0000lowastlowastlowast

Terpinolene 1290 1088 020 plusmn 003b 013 plusmn 001c 012 plusmn 001cd 062 plusmn 005a 3 5794 0000lowastlowastlowast

120573-Thujone 1430 1089 1716 plusmn 156c 2102 plusmn 232b 2209 plusmn 212a 672 plusmn 075d 3 5900699 0000lowastlowastlowast

120572-Thujone 1430 1089 020 plusmn 002b 007 plusmn 001c 028 plusmn 003a 004 plusmn 000d 3 1352 0001lowastlowastlowast

trans-Linalool oxide 1450 1088 003 plusmn 000c 002 plusmn 000d 604 plusmn 077a 258 plusmn 025b 3 1187662 0000lowastlowastlowast

trans-Sabinene hydrate 1474 1053 1207 plusmn 132c 1258 plusmn 130b 532 plusmn 056d 2178 plusmn 212a 3 2427412 0000lowastlowastlowast

cis-Linalool oxide 1478 1074 005 plusmn 000c 013 plusmn 001b 005 plusmn 001c 186 plusmn 016a 3 637193 0000lowastlowastlowast

120572-Copaene 1497 1379 050 plusmn 004b 407 plusmn 044a 033 plusmn 003c 022 plusmn 003d 3 356191 0000lowastlowastlowast

Camphor 1532 1143 002 plusmn 000d 004 plusmn 000c 015 plusmn 001a 012 plusmn 001b 3 1373 0001lowastlowastlowast

Linalool 1553 1088 200 plusmn 023a 080 plusmn 007b 009 plusmn 001d 015 plusmn 002c 3 932 0005lowastlowast

cis-Sabinene hydrate 1556 1082 002 plusmn 000d 182 plusmn 015a 180 plusmn 020b 080 plusmn 007c 3 44183 0000lowastlowastlowast

Linalyl acetate 1565 1239 050 plusmn 006b 458 plusmn 051a 008 plusmn 001d 021 plusmn 003c 3 530855 0000lowastlowastlowast

Bornyl acetate 1590 1270 110 plusmn 010a 004 plusmn 000c 036 plusmn 004b 002 plusmn 000d 3 26807 0000lowastlowastlowast

Terpinene-4-ol 1611 1176 300 plusmn 041a 015 plusmn 001c 005 plusmn 000d 110 plusmn 012b 3 2216 0000lowastlowastlowast

cis-Dihydrocarvone 1645 1193 048 plusmn 005b 094 plusmn 008a 011 plusmn 001c 003 plusmn 000d 3 9429 0000lowastlowastlowast

cis-Verbenol 1654 1132 234 plusmn 024b 002 plusmn 000d 031 plusmn 004c 357 plusmn 044a 3 1560473 0000lowastlowastlowast

Sabinyl acetate 1658 1291 029 plusmn 003b 501 plusmn 045a 011 plusmn 001c 008 plusmn 001d 3 1009581 0000lowastlowastlowast

trans-Sabinol 1666 1120 029 plusmn 003b 004 plusmn 000d 006 plusmn 001c 099 plusmn 008a 3 158213 0000lowastlowastlowast

Lavandulol 1677 1150 271 plusmn 031c 028 plusmn 003d 361 plusmn 034b 554 plusmn 061a 3 2600236 0000lowastlowastlowast

120572-Humulene 1687 1454 008 plusmn 001b 062 plusmn 005a 003 plusmn 000c 001 plusmn 000d 3 169933 0000lowastlowastlowast

Neral 1694 1240 001 plusmn 000d 026 plusmn 003c 268 plusmn 027a 028 plusmn 003b 3 1110653 0000lowastlowastlowast

120574-Muurolene 1692 1474 028 plusmn 003b 109 plusmn 011a 015 plusmn 002c 005 plusmn 000d 3 87132 0000lowastlowastlowast

120574-Curcumene 1704 1475 100 plusmn 012c 293 plusmn 032a 057 plusmn 006d 195 plusmn 022b 3 5219 0000lowastlowastlowast

120572-Terpinyl acetate 1705 1344 017 plusmn 001a tr 014 plusmn 001b 003 plusmn 000c 3 9111 0000lowastlowastlowast

120572-Terpineol 1706 1189 019 plusmn 002b 014 plusmn 001c 005 plusmn 001d 038 plusmn 004a 3 9312 0000lowastlowastlowast

Borneol 1719 1165 052 plusmn 006a 046 plusmn 005b 023 plusmn 003c tr 3 39735 0000lowastlowastlowast

Germacrene D 1726 1480 004 plusmn 000d 010 plusmn 001c 158 plusmn 017a 052 plusmn 006b 3 164756 0000lowastlowastlowast

Neryl acetate 1732 1356 020 plusmn 002c 026 plusmn 003b 003 plusmn 000d 051 plusmn 006a 3 4273 0000lowastlowastlowast

120573-Selinene 1742 1486 472 plusmn 051c 450 plusmn 050d 529 plusmn 062b 667 plusmn 074a 3 4548 0000lowastlowastlowast

120572-Bisabolene 1746 1494 088 plusmn 009b 011 plusmn 001c 125 plusmn 015a 002 plusmn 000d 3 1532 0001lowastlowastlowast

Carvone 1751 1242 200 plusmn 031a 014 plusmn 001c 046 plusmn 005b 005 plusmn 000d 3 988 0004lowastlowast

Geranyl acetate 1765 1383 008 plusmn 001b 061 plusmn 007a 003 plusmn 000c 002 plusmn 000d 3 2429 0000lowastlowastlowast

120574-Cadinene 1776 1526 040 plusmn 005c 112 plusmn 015b 127 plusmn 013a 005 plusmn 000d 3 9838 0000lowastlowastlowast

Nerol 1797 1228 010 plusmn 001c 011 plusmn 001c 033 plusmn 004b 193 plusmn 031a 3 211879 0000lowastlowastlowast

Geraniol 1857 1255 065 plusmn 007a 003 plusmn 000d 004 plusmn 000c 020 plusmn 002b 3 7796 0000lowastlowastlowast

p-Cymene-8-ol 1864 1183 tr 096 plusmn 008b 370 plusmn 041a tr 3 363671 0000lowastlowastlowast


Table 2 Continued


Jerissa Boukornine Kairouan Bou Salem120572-Calacorene 1920 1527 011 plusmn 001d 016 plusmn 002b 012 plusmn 001c 052 plusmn 006a 3 3553 0000lowastlowastlowast

Caryophyllene oxide 2008 1578 005 plusmn 000b 006 plusmn 001a 006 plusmn 000a tr 3 165 0000lowastlowastlowast

cis-Nerolidol 2050 1553 027 plusmn 003a 006 plusmn 001c 007 plusmn 001b 001 plusmn 000d 3 9288 0000lowastlowastlowast

Eugenol 2192 1356 015 plusmn 002d 124 plusmn 014a 032 plusmn 004b 020 plusmn 003c 3 21806 0000lowastlowastlowast

120572-Bisabolol 2232 1673 003 plusmn 000b 002 plusmn 000c 015 plusmn 002a 001 plusmn 000d 3 1907 0000lowastlowastlowast

Chamazulene 2430 1719 3993 plusmn 456a 2530 plusmn 312d 2951 plusmn 326c 3481 plusmn 411b 3 532471 0000lowastlowastlowast

Chemical classesMonoterpene hydrocarbons 5602 plusmn 621b 4531 plusmn 531d 4573 plusmn 522c 6208 plusmn 749a 3 2632256 0000lowastlowastlowast

Aromatic hydrocarbons 008 plusmn 001c 012 plusmn 001b 022 plusmn 003a 004 plusmn 000d 3 956342 0000lowastlowastlowast

Monoterpene alcohols 1222 plusmn 133b 429 plusmn 052d 886 plusmn 096c 1407 plusmn 167a 3 91267 0000lowastlowastlowast

Monoterpene esters 234 plusmn 031b 1005 plusmn 123a 075 plusmn 008d 087 plusmn 009c 3 56701 0000lowastlowastlowast

Monoterpene ketones 1986 plusmn 213c 2221 plusmn 211b 2309 plusmn 310a 696 plusmn 741d 3 15273 0000lowastlowastlowast

Monoterpene aldehydes 001 plusmn 000d 026 plusmn 003c 268 plusmn 245a 028 plusmn 003b 3 436002 0000lowastlowastlowast

Monoterpene ethers 011 plusmn 001d 018 plusmn 002c 644 plusmn 077a 452 plusmn 051b 3 426516 0000lowastlowastlowast

Sesquiterpenes 809 plusmn 099d 1478 plusmn 166a 1080 plusmn 145b 1002 plusmn 178c 3 123594 0000lowastlowastlowast

Total identified 9872 plusmn 1023b 9764 plusmn 978d 9855 plusmn 988c 9883 plusmn 976a 3 458123 0000lowastlowastlowast

RRI relative retention index a HP-INNOWax b HP-5 lowastCompounds in order of elution on HP-INNOWax values of volatile essential oil percentages are theaverage three determinations (119899 = 3)These values with different letters (andashd) are significantly different at 119875 lt 005 nd not detected NS not significant lowastlowast119875 lt001 lowastlowastlowast119875 lt 0001df degree of freedom 119865 Fisher value and 119875 probability

minus10minus08

minus06minus04

minus04

minus02

minus02

00

025035

045055

065075

085095

00

02

02

04

04

06

06

08

Axe 1(18

48)

Axe 2 (1104)

Axe

3(692

) Jeacuterissa

Boukornine

Kairouan

Bou Salem

Figure 2 Relative position of the regions of plant collection basedon their essential oils composition (Table 2) in the space defined bythe three principal components

showed that total polyphenol content of wormwood aerialparts collected in Romania was 9820mgg DW In additionEbrahimzadeh et al [45] reported that the total polyphenolcontent of Iranian wormwood aerial parts was 194 plusmn 97mgEAGg of extract which was significantly of greater valuein comparison with our results However a lower contentin total polyphenols (979mg EAGg extract) content wasdetermined by Sengul et al [46] Other herbs with high levels

of phenolics were Polygonum aviculare (112mg GAE100 gDW) and Valeriana officinalis (111mg GAE100 g DW) [47]In general phenolic compounds are known for their rolein the prevention of some diseases due to their richness inantioxidants and their antiradical effect protecting cellularcomponents against damage induced by free radicals Dueto the diversity of their chemical structures they are likelyto have very different antioxidant capacity [48] Polyphenoliccompounds are commonly found in both edible and inedibleplants and they have been reported to have multiple bio-logical effects including antioxidant activity [47] The basicflavonoids structure is the flavan nucleus which consistsof 15 carbon atoms arranged in three rings (C

6ndashC3ndashC6)

labeled A B and C Various classes of flavonoid differ in thelevel of oxidation and saturation of ring C while individualcompounds within a class differ in the substitution patternof rings A and B The differences in the structure andsubstitution will influence the phenoxyl radical stability andthereby the antioxidant properties of the flavonoids [49]

As for total polyphenols total flavonoids differ signif-icantly (119875 le 005) depending on the region but with amaximum in Kairouan region (12640 plusmn 232mg CEg DW)(Figure 3)

Our results are in agreement with those reported byWojdyło et al [47] in that the most common flavonoidsare mainly distributed in Compositae family On the otherhand A absinthium L aerial parts have been reported tocontain flavonoids [50ndash52] thymol and carvacrol as well asother phenolic compounds [53]These pharmacophores havebeen shown to possess potent antioxidant and free radicalscavenging activity [54] In the present study high levels ofphenolic and flavonoid contents were estimated For total


a

a

a

b

b

bc

c

c

dd d

0

20

40

60

80

100

120

140

160

180

TPP mg GAEg DW TF mg CEg DW TCT mg CEg DW

JeacuterissaBoukornineKairouan

Bou Salem

Figure 3 Total polyphenols (TPP) total flavonoids (TF) andtotal condensed tannin (TCT) contents of different regions of Aabsinthium L aerial parts GAE gallic acid equivalents CE catechinequivalents The letters (andashd) indicate significant differences (119875 lt005)

condensed tannins the maximum level was detected in theregion Boukornine (15235 plusmn 459mg CEg DW) followedby Jerissa (8350 plusmn 669mg CEg DW) Bou Salem (7394 plusmn219mgCEgDW) andKairouan (3719plusmn116mgCEgDW)regions (Figure 3)

34 Individual Phenolic Compounds For a clear understand-ing of the regional effect on metabolic changes especiallyon phenolics it is necessary to investigate the profiles ofindividual phenolic compounds in A absinthium L Hencechanges of these phenolic compounds were different at fourTunisian regions (Table 3) The obtained results show that allthe identified compounds were highly significant (119875 lt 0001)by the region effect Phenolic acidsmdashtannic gallic hemihy-drate chlorogenic caffeic vanillic syringic ferulicp-coumaric rosmarinic and trans-cinnamic acidsmdashrepre-sent themajor fraction of the polyphenols analyzed in regionsof Bou Salem (3496 plusmn 356) Kairouan (4557 plusmn 523)and Jerissa (5833 plusmn 621) while the flavones dominatedat Boukornine region (3861 plusmn 413) In the regions ofBou Salem Kairouan and Boukornine the flavone was thepredominant fraction (1944 plusmn 236 1188 plusmn 211 and3247 plusmn 423 resp) while in the region of Jerissa tannicacid was the predominant compound (3792 plusmn 402) Thequantification of total polyphenols of wormwood aerialparts by the Folin-Ciocalteu method gave different resultsin all studied regions In the region of Bou Salem totalpolyphenols measured by Folin-Ciocalteu method wereestimated to be 837mg GAEg DW and 2803 plusmn 312mggGAE DW by HPLC in Kairouan were 99 89mg GAEgDW by Folin-Ciocalteu and 1591 plusmn 165mg GAEg byHPLC in Boukornine were 7205mg GAEg DW by Folin-Ciocalteu and 4823 plusmn 521mg GAEg DM by HPLC andin Jerissa were 4939mg GAEg DW by Folin-Ciocalteu and8808plusmn759mgGAEg DWbyHPLCThese variations in thequantification showed a significant difference depending on

minus02

00

02

04

06

08

Axe

3(446

)

Jeacuterissa

Boukornine

Kairouan

Bou Salem

025

040

055

070

085

100

Axe 1(24

45)

minus04minus02

0002

0406

0810

12

Axe 2 (951)

Figure 4 Principal component analysis of different regions basedon the phenolic composition of A absinthium L (Table 3)

the method used An earlier study showed that the analysisby liquid chromatography (HPLC) revealed the presence ofother flavonoid glycosides (isoquercitrin quercetin-3-O-120573-D-glucoside quercetin-3-O-rhamnoglucoside isorham-netin-3-O-rhamnoglucoside and isorhamnetin-3-glucoside)[50] which are characterized by important antioxidantactivity Gallic acid widely used as an additive to preventfood spoilage is renowned for its anticarcinogenic anti-inflammatory and antimutagenic activities [55]

A principal component analysis (PCA) was performed inorder to discriminate between different regions of collectingof absinthe based on the phenolic composition This analysisrevealed the existence of one group composed by Boukor-nine Kairouan and Bou Salem regions But Jerissa is clearlydistinct from this group (Figure 4)

A absinthium L can be considered a good source ofnaturally occurring antioxidant compounds which have anapplication in food industry due to its phenolic composition

35 DPPH Test As indicated in Table 4 the methanolextracts are highly influenced by the regional effect (antiradi-cal activitywas region-dependent)Methanolic extract of BouSalem region shows the highest antioxidant activity (938 plusmn082mgmL) which was stronger than that of the positivecontrol BHT (1077 plusmn 298 120583gmL) The lower activity wasdetected in the region of Boukornine (4426 plusmn 192mgmL)Mahmoudi et al [9] have reported an antiradical of worm-wood aerial parts activity with an IC

50of 612 plusmn 306 120583gmL

this value was lower than our results Meanwhile Sengul etal [46] showed that the antiradical activity of the methanolextracts of the aerial parts of wormwood collected in Turkeywas 7178 as compared to the positive control (BHA =200mgL measuring 9321) Different results were foundby Wojdyło et al [47] who reported a highly significantpositive correlation (119877 = 08352 119875 lt 005) between


Table3Analysis

ofvaria

nce(ANOVA

)and

phenoliccompo

sitionin

(w

w)a

ndqu

antity(120583ggD

W)o

fmethano

lextractso

fthe

aeria

lpartsof

wormwoo

d(A

absinthium

L)

Com

poun

dCollectionregion

df119865

119875

BouSalem

Kairo

uan

Jeris

saBo

ukornine

120583ggD

W

120583ggD

W

120583ggD

W

120583ggD

W

Phenolicacids

Tann

icacid

037plusmn004

D13

2plusmn011

d073plusmn008

C459plusmn051

b334plusmn345

A3792plusmn402

a19

9plusmn201

B413plusmn044

c3

2028659

000

0lowastlowastlowast

Gallic

acid

038plusmn004

D13

6plusmn012

c041plusmn003

C258plusmn026

b475plusmn055

A539plusmn061

a063plusmn007

B13

1plusmn011

d3

2925813

000

0lowastlowastlowast

Chlorogenica

cidhemihydrate

336plusmn042

A1199plusmn13

2a14

9plusmn016

C93

7plusmn098

b233plusmn031

B265plusmn031

c081plusmn009

D16

8plusmn017

d3

898916

2000

0lowastlowastlowast

Caffeicacid

ndnd

152plusmn016

A95

5plusmn099

a14

3plusmn013

B16

2plusmn014

bnd

nd3

8639266

000

0lowastlowastlowast

Vanillica

cid

047plusmn005

B16

8plusmn015

b094plusmn008

A591plusmn066

and

Nd

ndnd

31867878

000

0lowastlowastlowast

Syrin

gica

cid

ndnd

ndnd

126plusmn014

143plusmn012

ndnd

3681633

000

0lowastlowastlowast

Ferulic

acid

035plusmn004

D12

5plusmn011

d068plusmn007

B427plusmn044

a117plusmn015

A13

3plusmn010

c065plusmn007

C13

5plusmn012

b3

417574

000

0lowastlowastlowast

p-Cou

maricacid

306plusmn041

A1092plusmn12

3a055plusmn006

D346plusmn041

b217plusmn023

B246plusmn025

c059plusmn006

C12

2plusmn011

d3

4768092

000

0lowastlowastlowast

Rosm

arinicacid

091plusmn008

C325plusmn041

a033plusmn004

D207plusmn022

d252plusmn026

A286plusmn030

b13

0plusmn014

B270plusmn028

c3

2175

6000

0lowastlowastlowast

trans-C

innamicacid

090plusmn008

C321plusmn034

c060plusmn005

D377plusmn036

a235plusmn025

A267plusmn024

d16

9plusmn017

B350plusmn036

b3

1060

0003lowastlowast

Total

980plusmn088

B34

96plusmn356

c72

5plusmn081

D45

57plusmn523

b5138plusmn436

A58

33plusmn621

a76

6plusmn080

C1588plusmn16

6dFlavanols

Catechin

026plusmn003

C093plusmn008

c050plusmn004

B314plusmn044

b457plusmn050

A519plusmn056

and

nd3

589952

000

0lowastlowastlowast

Epicatechin

067plusmn008

239plusmn024

ndnd

ndNd

ndnd

31713630

000

0lowastlowastlowast

Total

093plusmn008

B332plusmn035

b050plusmn006

C314plusmn029

c457plusmn052

A519plusmn062

and

ndFlavanon

esNaringenin

120plusmn011

C428plusmn051

b115plusmn012

D72

3plusmn077

a211plusmn022

A240plusmn025

d13

1plusmn014

B272plusmn028

c3

3614

2000

0lowastlowastlowast

Total

120plusmn010

C428plusmn045

b11

5plusmn010

D72

3plusmn077

a211plusmn023

A240plusmn026

d13

1plusmn012

B272plusmn028

c

Flavon

ols

Myristin

087plusmn009

C310plusmn033

d059plusmn006

D371plusmn036

c345plusmn041

A392plusmn041

b304plusmn040

B630plusmn074

a3

23583

000

0lowastlowastlowast

Quercetin

dihydrate

094plusmn008

C33

5plusmn035

a033plusmn004

D207plusmn022

b10

6plusmn010

B12

0plusmn013

c16

1plusmn017

A334plusmn036

a3

29686

000

0lowastlowastlowast

Camph

erol

057plusmn006

D203plusmn025

c12

6plusmn013

C79

2plusmn084

a13

5plusmn014

B15

3plusmn016

d15

5plusmn016

A321plusmn034

b3

4442309

000

0lowastlowastlowast

Total

238plusmn024

C849plusmn099

c218plusmn022

D1370plusmn14

5a586plusmn066

B665plusmn076

d620plusmn065

A1286plusmn14

2bFlavon

esAp

igenin

140plusmn013

C499plusmn052

b041plusmn005

D258plusmn026

d278plusmn029

B316plusmn041

c296plusmn033

A614plusmn072

a3

129479

6000

0lowastlowastlowast

Flavon

e545plusmn061

C1944plusmn236

b18

9plusmn017

D1188plusmn211

c601plusmn059

B682plusmn073

d1566plusmn17

2A3247plusmn423

a3

5334547

000

0lowastlowastlowast

Total

738plusmn088

C26

33plusmn277

b230plusmn022

D1446plusmn16

6c879plusmn099

B998plusmn12

3d1862plusmn201

A38

61plusmn

413

a

Cou

marins

Cou

marin

459plusmn051

B1638plusmn17

8b16

7plusmn017

C1050plusmn15

6d1334plusmn114A

1515plusmn17

9c1331plusmn15

1A2760plusmn286

a3

61887

000

0lowastlowastlowast

Total

459plusmn055

B1638plusmn18

8b16

7plusmn018

C1050plusmn12

3d1334plusmn16

2A1515plusmn16

2c1331plusmn14

3A2760plusmn297

a

Total

2803plusmn312

C100plusmn92

3a1591plusmn

165D

100plusmn846

a88

08plusmn759A

100plusmn98

7a48

23plusmn521

B100plusmn899

a

Thev

alueso

fthe

levelsandpercentageso

fpheno

liccompo

unds

representthe

averageo

fthree

replicates

(119899=3)n

dno

tdetectedLette

rs(andashd

(lower

case)betweenpercentagesa

ndAndash

D(capita

lletters)b

etween

amou

nts)indicatesig

nificantd

ifferencesa

t119875lt005lowastlowastlowastSign

ificant

effectat119875lt0001dfdegree

offre

edom

119865Fish

ervalueand119875probability


Table 4 DPPH test (IC50 120583gmL) and reducing power (EC50 120583gmL) of methanol extracts of wormwood aerial parts

Bou Salem Kairouan Jerissa Boukornine BHT Ascorbic acid df 119865 119875

DPPH (IC50 120583gmL) 938 plusmn 082d 1899 plusmn 038c 3174 plusmn 123b 4426 plusmn 192a 1077 plusmn 298 3 5646 0000lowastlowastlowast

Reducing power(EC50 120583gmL) 236 plusmn 015b 403 plusmn 045d 246 plusmn 046a 216 plusmn 005c 420 plusmn 011 3 127 0348NS

IC50 and EC50 values represent the mean of three replicates (119899 = 3) letters (andashd) indicate significant differences at 119875 lt 005 lowastlowastlowastSignificant at 119875 lt 0001 NSnot significant df degree of freedom 119865 Fisher value and 119875 probability

Table 5 Antibacterial (IZ mm) and antifungal (119868) activities of wormwood essential oil

Boukornine Jerissa Kairouan Bou Salem df 119865 119875 Tetracycline NystatinBacteria

Staphylococcus aureus25923 2066 plusmn 261b 2066 plusmn 065b 25 plusmn 113a 18 plusmn 113c 3 205 0184NS 25 plusmn 256

Staphylococcusmethicillin-resistant 1500 plusmn 113b 1433 plusmn 065c 16 plusmn 130a 1433 plusmn 065c 3 021 0879NS 25 plusmn 254

ListeriamonocytogenesATCC 19195 1866 plusmn 235b 1733 plusmn 172c 2000 plusmn 195a 2000 plusmn 113a 3 053 0674NS 24 plusmn 242

FungiFusarium graminearum 1466 plusmn 057c 2365 plusmn 057a 1111 plusmn 012d 1666 plusmn 057b 3 1857 0000lowastlowastlowast 25 plusmn 231Fusarium culmorum 4523 plusmn 311b 2365 plusmn 057d 4206 plusmn 057c 4682 plusmn 057a 3 3277 0000lowastlowastlowast 25 plusmn 214Fusarium oxysporum 1904 plusmn 057c 2758 plusmn 255a 1794 plusmn 057d 2528 plusmn 057b 3 779 0009lowastlowast 20 plusmn 167Sclerotinia nd 2361 plusmn 212 nd nd 3 18581 0000lowastlowastlowast 22 plusmn 201Rhizoctonia solani nd 2539 plusmn 057 nd nd 3 7735 0000lowastlowastlowast 20 plusmn 178Results are the mean of three replications The diameter of disc was 6mm Values with different superscripts (andashd) are significantly different at 119875 lt 005 ndnot detected NS not significant df degree of freedom 119865 Fisher value and 119875 probability lowastlowast119875 lt 001 lowastlowastlowast119875 lt 0001

the antiradical activity of Lamiaceae and total polyphenolsdemonstrating the importance of these compounds in theantioxidant activity of extracts of spices and their significantcontribution to the total antioxidant activity

36 Reducing Power The ability of methanol extracts ofwormwood aerial parts collected from four regions to reducethe metal ions was evaluated by determining the CE

50corre-

sponding to the concentration of the extract to an absorbanceof 05 The ascorbic acid has been used as a positive controlThe results in Table 4 showed that the methanol extracts ofwormwood aerial parts were characterized by EC

50values

lower than that of the positive control ascorbic acid (EC50=

42plusmn011 120583gmL) In addition statistical analysis showed thatthere is no effect of region on the activities of the methanolextracts

37 Antibacterial Activity The test results of the antibacterialeffect are summarized in Table 5 The results show thatthe diameter of the inhibition zone is not affected by theregion factor The essential oils of wormwood at differentregions showed an interesting antibacterial activity Indeedthe highest activity was observed against Staphylococcusaureus strain with a diameter of inhibition equal to 25 plusmn113mm for the essential oil of Kairouan this strain had alower sensitivity to the EO of Bou Salem region (IZ = 18 plusmn113mm) Essential oils of Boukornine and Jerissa regionsshow a moderately antibacterial activity (IZ = 2066mm) Inagreement with our results the EO of wormwood aerial parts

growing in Canada has a high activity against Staphylococcusaureus (IZ = 25 plusmn 14mm) which was greater than thepositive control (methicillin activity IZ = 18 plusmn 10mm) [34]Essential oils of Bou Salem and Kairouan have the sameactivity against Listeria monocytogenes (IZ = 20mm) but theEO of Jerissa and Boukornine is less active (IZ = 18mm)These results show that wormwood essential oil was endowedwith a significant antibacterial activity that is closely relatedto the organoleptic quality of the oil which in turn dependsstrongly on the collection region of plant material

38 Antifungal Activity The results of the antifungal activityof essential oils of absinthe are summarized in Table 5Wormwood EOs had a significant inhibitory activity againstthe three phytopathogenic F graminearum F culmorum andF oxysporum while only EO of Jerissa proved to be activeagainst Sclerotinia (2361 plusmn 212) and Rhizoctonia solani(2539 plusmn 057) As for the antibacterial activity antifungalactivity could be attributed to the major component ofessential oil of wormwood chamazulene [56] Meanwhile ithas been shown that the chamazulene possesses significantantifungal activity [57] The antimicrobial effect of EOs ofabsinthe gives them an important role in the fields of foodcosmetics and pharmaceutical industries In addition Aabsinthium L EO from France containing (Z)-epoxyocimeneand chrysanthenyl acetate as major components inhibitedthe growth of both the yeasts Candida albicans and Sac-charomyces cerevisiae var chevalieri [4] Wormwood EOfrom a Turkish population whose main components are


camphor 18-cineole and chamazulene has been describedas fungicidal against 34 species of fungi including F solaniand F oxysporum [3] Furthermore A absinthium EO fromUruguay rich in thujone showed antifungal effects againstAlternaria sp and Botrytis cinerea [58] However furtherstudies are needed to identify the compound(s) responsiblefor the antifungal effects of A absinthium L EOs

Conflict of Interests

The authors declare that there is no conflict of interests

References

[1] K S Bora and A Sharma ldquoThe genus Artemisia a comprehen-sive reviewrdquo Pharmaceutical Biology vol 49 no 1 pp 101ndash1092011

[2] K S Bora andA Sharma ldquoPhytochemical and pharmacologicalpotential of Artemisia absinthium Linn and Artemisia asiaticaNakai a reviewrdquo Journal of Pharmacy Research vol 3 no 2 pp325ndash328 2010

[3] S Kordali R Kotan A Mavi A Cakir A Ala and AYildirim ldquoDetermination of the chemical composition andantioxidant activity of the essential oil of Artemisia dracunculusand of the antifungal and antibacterial activities of TurkishArtemisia absinthium A dracunculus Artemisia santonicumand Artemisia spicigera essential oilsrdquo Journal of Agriculturaland Food Chemistry vol 53 no 24 pp 9452ndash9458 2005

[4] F Juteau I Jerkovic VMasotti et al ldquoComposition and antimi-crobial activity of the essential oil of Artemisia absinthium fromCroatia and Francerdquo Planta Medica vol 69 no 2 pp 158ndash1612003

[5] H Chiasson A Belanger N Bostanian C Vincent and APoliquin ldquoAcaricidal properties of Artemisia absinthium andTanacetum vulgare (Asteraceae) essential oils obtained by threemethods of extractionrdquo Journal of Economic Entomology vol94 no 1 pp 167ndash171 2001

[6] K A Tariq M Z Chishti F Ahmad and A S ShawlldquoAnthelmintic activity of extracts of Artemisia absinthiumagainst ovine nematodesrdquo Veterinary Parasitology vol 160 no1-2 pp 83ndash88 2009

[7] S Irshad M Butt and Y Hira ldquoIn-vitro antibacterial activityof twomedicinal plants neem (Azadirachta indica) and pepper-mintrdquo International Research Journal of Pharmaceuticals vol 1no 1 pp 9ndash14 2011

[8] A-U H Gilani and K H Janbaz ldquoPreventive and curativeeffects of Artemisia absinthium on acetaminophen and CCl4-induced hepatotoxicityrdquo General Pharmacology vol 26 no 2pp 309ndash315 1995

[9] MMahmoudiM A Ebrahimzadeh F Ansaroudi S F Nabaviand S M Nabavi ldquoAntidepressant and antioxidant activities ofArtemisia absinthium L at flowering stagerdquo African Journal ofBiotechnology vol 8 no 24 pp 7170ndash7175 2009

[10] J M Canadanovic-Brunet S M Djilas G S Cetkovic andV T Tumbas ldquoFree-radical scavenging activity of wormwood(Artemisia absinthium L) extractsrdquo Journal of the Science of Foodand Agriculture vol 85 no 2 pp 265ndash272 2005

[11] O Erturk and U Uslu ldquoAntifeedant growth and toxic effectsof some plant extracts on Leptinotarsa decemlineata (Say)(Coleoptera Chrysomelidae)rdquo Fresenius Environmental Bul-letin vol 16 no 6 pp 601ndash607 2007

[12] T Jiratanan and R H Liut ldquoAntioxidant activity of processedtable beets (Beta vulgaris var conditiva) and green beans(Phaseolus vulgaris L)rdquo Journal of Agricultural and Food Chem-istry vol 52 no 9 pp 2659ndash2670 2004

[13] J N Losso F Shahidi and D Bagchi Anti-Angiogenic Func-tional and Medicinal Foods Taylor amp Francis Boca Raton FlaUSA 2007

[14] B N Singh B R Singh R L Singh et al ldquoOxidative DNAdam-age protective activity antioxidant and anti-quorum sensingpotentials of Moringa oleiferardquo Food and Chemical Toxicologyvol 47 no 6 pp 1109ndash1116 2009

[15] A Basile S Sorbo B Conte et al ldquoAntioxidant activity inextracts from Leptodictyum riparium (Bryophyta) stressed byheavy metals heat shock and salinityrdquo Plant Biosystems vol145 no 1 pp 77ndash80 2011

[16] M Minutolo I Caruso G Caruso P Chiaiese and A ErricoldquoEstablishment of Aster sedifolius and Aster caucasicus calluscultures as a potential source of antioxidantsrdquo Plant Biosystemsvol 146 no 1 pp 41ndash46 2012

[17] P-G Pietta ldquoFlavonoids as antioxidantsrdquo Journal of NaturalProducts vol 63 no 7 pp 1035ndash1042 2000

[18] M Friedman P R Henika and R E Mandrell ldquoBactericidalactivities of plant essential oils and some of their isolatedconstituents against Campylobacter jejuni Escherichia coli Lis-teria monocytogenes and Salmonella entericardquo Journal of FoodProtection vol 65 no 10 pp 1545ndash1560 2002

[19] K M Schuenzel and M A Harrison ldquoMicrobial antagonistsof foodborne pathogens on fresh minimally processed vegeta-blesrdquo Journal of Food Protection vol 65 no 12 pp 1909ndash19152002

[20] M Sokmen J Serkedjieva D Daferera et al ldquoIn vitro antiox-idant antimicrobial and antiviral activities of the essential oiland various extracts from herbal parts and callus cultures ofOriganum acutidensrdquo Journal of Agricultural and Food Chem-istry vol 52 no 11 pp 3309ndash3312 2004

[21] CWWright Artemisia Taylor amp Francis New York NY USA2002

[22] A Rezaeinodehi and S Khangholi ldquoChemical composition ofthe essential oil of Artemisia absinthium growing wild in IranrdquoPakistan Journal of Biological Sciences vol 11 no 6 pp 946ndash9492008

[23] E Derwich Z Benziane and A Boukir ldquoChemical composi-tions and insectisidal activity of essential oils of three plantsArtemisia SP Artemisia herba-alba Artemisia absinthium andArtemisia Pontica (Morocco)rdquo Electronic Journal of Environ-mental Agricultural and Food Chemistry vol 8 no 11 pp 1202ndash1211 2009

[24] L Martın A M Mainar A Gonzalez-Coloma J Burilloand J S Urieta ldquoSupercritical fluid extraction of wormwood(Artemisia absinthium L)rdquo Journal of Supercritical Fluids vol56 no 1 pp 64ndash71 2011

[25] Council of Europe European Pharmacopoeia Council ofEurope Strasbourg France 3rd edition 1997

[26] J-L Mau C-N Chang S-J Huang and C-C Chen ldquoAntiox-idant properties of methanolic extracts from Grifola frondosaMorchella esculenta and Termitomyces albuminosus myceliardquoFood Chemistry vol 87 no 1 pp 111ndash118 2004

[27] V Dewanto W Xianzhong K K Adom and R H LiuldquoThermal processing enhances the nutritional value of tomatoesby increasing total antioxidant activityrdquo Journal of Agriculturaland Food Chemistry vol 50 no 10 pp 3010ndash3014 2002


[28] B Sun J M Ricardo-da-Silva and I Spranger ldquoCritical factorsof vanillin assay for catechins and proanthocyanidinsrdquo Journalof Agricultural and Food Chemistry vol 46 no 10 pp 4267ndash4274 1998

[29] T Hatano H Kagawa T Yasuhara and T Okuda ldquoTwonew flavonoids and other constituents in licorice root theirrelative astringency and radical scavenging effectsrdquo Chemicaland Pharmaceutical Bulletin vol 36 no 6 pp 2090ndash2097 1988

[30] M Oyaizu ldquoStudies on products of browning reaction antiox-idative activity of products of browning reactionrdquoThe JapaneseJournal of Nutrition vol 44 no 6 pp 307ndash315 1986

[31] J L Rıos and M C Recio ldquoMedicinal plants and antimicrobialactivityrdquo Journal of Ethnopharmacology vol 100 no 1-2 pp 80ndash84 2005

[32] Statsoft STATISTICA for Windows (Computer Program Elec-tronic 703 Manual) Statsoft Inc Tulsa Okla USA 1998

[33] A Orav A Raal E Arak M Muurisepp and T KailasldquoComposition of the essential oil of Artemisia absinthium Lof different geographical originrdquo Proceedings of the EstonianAcademy of Sciences Chemistry vol 55 no 3 pp 155ndash165 2006

[34] D Lopes-Lutz D S Alviano C S Alviano and P PKolodziejczyk ldquoScreening of chemical composition antimi-crobial and antioxidant activities of Artemisia essential oilsrdquoPhytochemistry vol 69 no 8 pp 1732ndash1738 2008

[35] KMsaada KHosniM B Taarit T ChahedM EKchouk andBMarzouk ldquoChanges on essential oil composition of coriander(Coriandrum sativum L) fruits during three stages of maturityrdquoFood Chemistry vol 102 no 4 pp 1131ndash1134 2007

[36] K Msaada M B Taarit K Hosni M Hammami and BMarzouk ldquoRegional and maturational effects on essential oilsyields and composition of coriander (Coriandrum sativum L)fruitsrdquo Scientia Horticulturae vol 122 no 1 pp 116ndash124 2009

[37] KMsaada KHosniM B Taarit O Ouchikh and BMarzoukldquoVariations in essential oil composition during maturation ofcoriander (Coriandrum sativum l) Fruitsrdquo Journal of FoodBiochemistry vol 33 no 5 pp 603ndash612 2009

[38] K Masada K Hosni M Ben Taarit M Hammami and BMarzouk ldquoEffects of crop season and maturity stages on theyield and composition of essential oil of coriander (Coriandrumsativum L) fruitrdquo Medicinal and Aromatic Plant Science andBiotechnology vol 6 no 1 pp 115ndash122 2012

[39] K Msaada M B Taarit K Hosni et al ldquoComparison of differ-ent extraction methods for the determination of essential oilsand related compounds from coriander (Coriandrum sativumL)rdquo Acta Chimica Slovenica vol 59 no 4 pp 803ndash813 2012

[40] S Kordali A Cakir AMavi H Kilic and A Yildirim ldquoScreen-ing of chemical composition and antifungal and antioxidantactivities of the essential oils from three Turkish Artemisiaspeciesrdquo Journal of Agricultural and Food Chemistry vol 53 no5 pp 1408ndash1416 2005

[41] B M Lawrence ldquoProgress in essential oilsrdquo Perfumer amp Fla-vorist vol 17 pp 39ndash42 1992

[42] W C Evans Trease and EvansPharmacognosy Saunders Edin-burgh UK 15th edition 2000

[43] N Salem KMsaada G Hamdaoui F Limam and BMarzoukldquoVariation in phenolic composition and antioxidant activityduring flower development of safflower (Carthamus tinctoriusL)rdquo Journal of Agricultural and Food Chemistry vol 59 no 9pp 4455ndash4463 2011

[44] H Monica A Moisuc F Radu S Dragan and I GergenldquoTotal polyphenols content determination in complexmatrix of

medicinal plants fromRomania by nir spectroscopyrdquo Bulletin ofthe University of Agricultural Sciences and Veterinary MedicineCluj-Napoca Agriculture vol 65 no 1 pp 123ndash128 2008

[45] M A Ebrahimzadeh S F Nabavi S M Nabavi and FPourmorad ldquoNitric oxide radical scavenging potential of someElburz medicinal plantsrdquo African Journal of Biotechnology vol9 no 32 pp 5212ndash5217 2010

[46] M Sengul S Ercisli H Yildiz N Gungor A Kavaz andB Cetina ldquoAntioxidant antimicrobial activity and total phe-nolic content within the aerial parts of artemisia absinthumArtemisia santonicum and Saponaria officinalisrdquo Iranian Journalof Pharmaceutical Research vol 10 no 1 pp 49ndash56 2011

[47] A Wojdyło J Oszmianski and R Czemerys ldquoAntioxidantactivity and phenolic compounds in 32 selected herbsrdquo FoodChemistry vol 105 no 3 pp 940ndash949 2007

[48] A Dapkevicius T A Van Beek G P Lelyveld et al ldquoIsolationand structure elucidation of radical scavengers from Thymusvulgaris leavesrdquo Journal of Natural Products vol 65 no 6 pp892ndash896 2002

[49] K Aaby E Hvattum and G Skrede ldquoAnalysis of flavonoidsand other phenolic compounds using high-performance liquidchromatography with coulometric array detection relationshipto antioxidant activityrdquo Journal of Agricultural and Food Chem-istry vol 52 no 15 pp 4595ndash4603 2004

[50] G-Q Zheng ldquoCytotoxic terpenoids and flavonoids fromArtemisia annuardquo Planta Medica vol 60 no 1 pp 54ndash57 1994

[51] C A Rice-Evans N J Miller and G Paganga ldquoStructure-antioxidant activity relationships of flavonoids and phenolicacidsrdquo Free Radical Biology andMedicine vol 20 no 7 pp 933ndash956 1996

[52] H-G Lee H Kim W-K Oh et al ldquoTetramethoxy hydrox-yflavone p7F downregulates inflammatory mediators via theinhibition of nuclear factor120581BrdquoAnnals of theNewYorkAcademyof Sciences vol 1030 pp 555ndash568 2004

[53] S Kordali I Aslan O Calmasur and A Cakir ldquoToxicityof essential oils isolated from three Artemisia species andsome of their major components to granary weevil Sitophilusgranarius (L) (Coleoptera Curculionidae)rdquo Industrial Cropsand Products vol 23 no 2 pp 162ndash170 2006

[54] A Braca G Fico I Morelli F de Simone F Tome and N DeTommasi ldquoAntioxidant and free radical scavenging activity offlavonol glycosides from different Aconitum speciesrdquo Journal ofEthnopharmacology vol 86 no 1 pp 63ndash67 2003

[55] Y-Y Soong and P J Barlow ldquoQuantification of gallic acidand ellagic acid from longan (Dimocarpus longan Lour) seedand mango (Mangifera indica L) kernel and their effects onantioxidant activityrdquo Food Chemistry vol 97 no 3 pp 524ndash5302006

[56] B Bozin N Mimica-Dukic M Bogavac et al ldquoChemical com-position antioxidant and antibacterial properties of AchilleacollinaBecker exHeimerl sl andA pannonica Scheele essentialoilsrdquoMolecules vol 13 no 9 pp 2058ndash2068 2008

[57] B Kedzia ldquoAntimicroorganisms activity of oilChamomillae andits componentsrdquo Herba Polonica vol 37 no 1 pp 29ndash38 1991

[58] M L Umpierrez M E Lagreca R Cabrera G Grille and CRossini ldquoEssential oils from Asteraceae as potential biocontroltools for tomato pests and diseasesrdquo Phytochemistry Reviewsvol 11 no 4 pp 339ndash350 2012

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy


Carbohydrate Chemistry

International Journal of


Journal of

Chemistry


Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of


Chromatography Research International


Applied ChemistryJournal of



Theoretical ChemistryJournal of


Journal of

Spectroscopy

Analytical ChemistryInternational Journal of


Journal of


Quantum Chemistry


Organic Chemistry International

ElectrochemistryInternational Journal of



CatalystsJournal of


the large scale production of absinthe began In additionArtemisia absinthium L was used freshly for the plant speciesas well as for the alcoholic beverage

Aromatic and medicinal plants are an easily accessibleand edible source of natural antioxidants They have theability to protect the organism from damage caused byfree radical-induced oxidative stress such as cancer andcardiovascular and neurodegenerative diseases [12 13]

Various studies have reported previously that reactiveoxygen species (ROS) are highly reactive and cause damageto protein lipids enzymes and DNA [14] In this contextthe use of plants and herbs in food pharmaceutical andcosmetic industries as a source of natural antioxidant andbiologically active compounds [15 16] has attracted a greatdeal of scientific interest In addition Pietta [17] reported thatthe antioxidant effect of plant products is mainly attributed tophenolic compounds

In the food industry a large spectrum of microorgan-ism leads to food spoilage Therefore preservation of foodmaterial from degradation mainly by oxidation processesand microorganism activity during production storage andmarketing is an important stage Also Friedman et al[18] reported the increase of some pathogensrsquo resistance tosynthetic antibiotics which are also uncomfortable to patientsdue to their adverse drug reactions

That is why the use of plants and herbs as a sourceof natural products has attracted the interest of manyresearchers They have been screened for their potential usesas alternative remedies for the treatment of many infectionsas an antioxidant and also as a natural food preservative[19 20]

Many authors have reported composition and antioxidantand antibacterial proprieties of A absinthium L essentialoil and extracts Wright [21] has reported that wormwoodstimulant propriety is caused by bitter substances as artabsin(sesquiterpene lactone) and absinthin (dimer of sesquiter-pene lactone) present in plant extracts In fact Iranian worm-wood essential oil was characterized by the predominance of120573-pinene and 120573-thujone [22] In the same context essentialoil of A absinthium L collected from Morocco is essentiallyrepresented mainly by 120572-thujone (3969) sabinyl acetate(1096) and 120573-thujone (725) [23] Furthermore Martınet al [24] showed that the major compounds of wormwoodfound in the SFE extracts as well as in the hydrodistilledessential oils were Z-epoxyocimene chrysanthenol andchrysanthenyl acetate

To the best of our knowledge there are no reportsdealing with chemotype A absinthium L essential oil inTunisia In our investigation we have studied essential oiland methanolic extracts composition of wild populationof wormwood cultivated in Tunisia and harvested fromfour distinct regions (Jerissa Bou Salem Boukornine andKairouan) Furthermore we have evaluated their antioxidantantibacterial and antifungal activities The purpose of thisstudy was to underline the variability of wormwood essentialoil and methanolic extracts and their biological activities asaffected by the collection site

2 Material and Methods

21 Chemicals All solvents used in the experiments (diethylether acetonitrile and water of high-performance liquidchromatography (HPLC) grade and ethanol methanoland ethyl acetate of analytical grade) were purchasedfrom Merck (Darmstadt Germany) Sodium hydroxide(NaOH) sodium phosphate (Na

2HPO4) sodiummonobasic

phosphate anhydrous (NaH2PO4H2O) sodium carbonate

(Na2CO3) sodium nitrite NaNO

2 butylated hydroxytoluene

(BHT) 120573-carotene linoleic acid ethylenediaminetetraaceticacid (EDTA) 3-(2-pyridyl)-56-bis(4-phenyl-sulphonicacid)-124-triazine (ferrozine) iron(II) chloride tetrahydrate(FeCl2sdot4H2O) iron(II) chloride (FeCl

2) iron(III) chloride

(FeCl3) 11-diphenyl-2-picrylhydrazyl (DPPH) polyvinyl-

polypyrrolidone Folin-Ciocalteu reagent potassium ferri-cyanide (K

3Fe(CN)

6) aluminium chloride (AlCl

3) homolo-

gous series of C6ndashC17

n-alkanes and high-purity standardsof essential oil were purchased from Sigma-Aldrich(Steinheim Germany) Hydrochloric acid (HCl) (36ndash38) and potassium hydroxide (KOH) were provided by JT Baker (Deventer Netherlands) Trifluoroacetic acid (TFA)(999) was purchased from ROMIL Ltd (Cambridge UK)Cyanidin 3-O-glucoside was purchased from Extrasynthese(Genay France) Deionized water was used to prepare allsolutions unless otherwise indicated These solutions werewrapped in aluminium foil and stored at 4∘C All otherchemicals used were of analytical grade

22 Plant Material A absinthium L aerial parts were har-vested randomly at flowering stage from different Tunisianregions The choice of these sites was dictated by the geo-graphical differences of the species studied (Table 1) Theplant material was harvested from four regions (Jerissa BouSalem Boukornine andKairouan)Theharvested aerial partswere identified by Professor Abderrazek Smaoui (Borj CedriaBiotechnology Center Tunisia) according to the Tunisianflora and a voucher specimen was deposited at the herbariumof the Laboratory of Bioactive Substances (BiotechnologyCenter of Borj Cedria) under the number Aab21213 Theharvested material was freeze-dried and then ground to finepowder by an electric mill and conserved in a dessicator atroom temperature (sim25∘C) in darkness for further uses

23 Essential Oil Extraction The leaves stems and flowerswere cut into small pieces and subjected to hydrodistillationtype Clevenger for 90min in accordance with EuropeanPharmacopoeia method [25] Essential oil extractions weredone in triplicate for each A absinthium L collection siteYield percentagewas calculated asmg of essential oil per 100 gof plant dry matter All experiments were done in triplicate

24 Gas Chromatography (GC) Analysis Analysis of Aabsinthium L essential oil volatile compounds by gas chro-matography (GC) was carried out on a Hewlett-Packard6890 gas chromatograph (Palo Alto CA USA) equippedwith a flame ionization detector (FID) and an electronic





2) was


6ndashC22








2SO4were














119860blank] times 100 (1)


50) was



3Fe (CN)

6) and incubated


50) was









a

bc

bc

0

02

04

06

08

1

12

14

16

18


Esse

ntia

l oil

yiel

d (

ww

)


































































Table 2 Continued


















minus10minus08

minus06minus04

minus04

minus02

minus02

00

025035

045055

065075

085095

00

02

02

04

04

06

06

08

Axe 1(18

48)

Axe 2 (1104)

Axe

3(692

) Jeacuterissa

Boukornine

Kairouan

Bou Salem




6ndashC3ndashC6)





a

a

a

b

b

bc

c

c

dd d

0

20

40

60

80

100

120

140

160

180



Bou Salem




minus02

00

02

04

06

08

Axe

3(446

)

Jeacuterissa

Boukornine

Kairouan

Bou Salem

025

040

055

070

085

100

Axe 1(24

45)

minus04minus02

0002

0406

0810

12

Axe 2 (951)






50of 612 plusmn 306 120583gmL



Table3Analysis

ofvaria

nce(ANOVA

)and

phenoliccompo

sitionin

(w

w)a

ndqu

antity(120583ggD

W)o

fmethano

lextractso

fthe

aeria

lpartsof

wormwoo

d(A

absinthium

L)

Com

poun

dCollectionregion

df119865

119875

BouSalem

Kairo

uan

Jeris

saBo

ukornine

120583ggD

W

120583ggD

W

120583ggD

W

120583ggD

W

Phenolicacids

Tann

icacid

037plusmn004

D13

2plusmn011

d073plusmn008

C459plusmn051

b334plusmn345

A3792plusmn402

a19

9plusmn201

B413plusmn044

c3

2028659

000

0lowastlowastlowast

Gallic

acid

038plusmn004

D13

6plusmn012

c041plusmn003

C258plusmn026

b475plusmn055

A539plusmn061

a063plusmn007

B13

1plusmn011

d3

2925813

000

0lowastlowastlowast

Chlorogenica

cidhemihydrate

336plusmn042

A1199plusmn13

2a14

9plusmn016

C93

7plusmn098

b233plusmn031

B265plusmn031

c081plusmn009

D16

8plusmn017

d3

898916

2000

0lowastlowastlowast

Caffeicacid

ndnd

152plusmn016

A95

5plusmn099

a14

3plusmn013

B16

2plusmn014

bnd

nd3

8639266

000

0lowastlowastlowast

Vanillica

cid

047plusmn005

B16

8plusmn015

b094plusmn008

A591plusmn066

and

Nd

ndnd

31867878

000

0lowastlowastlowast

Syrin

gica

cid

ndnd

ndnd

126plusmn014

143plusmn012

ndnd

3681633

000

0lowastlowastlowast

Ferulic

acid

035plusmn004

D12

5plusmn011

d068plusmn007

B427plusmn044

a117plusmn015

A13

3plusmn010

c065plusmn007

C13

5plusmn012

b3

417574

000

0lowastlowastlowast

p-Cou

maricacid

306plusmn041

A1092plusmn12

3a055plusmn006

D346plusmn041

b217plusmn023

B246plusmn025

c059plusmn006

C12

2plusmn011

d3

4768092

000

0lowastlowastlowast

Rosm

arinicacid

091plusmn008

C325plusmn041

a033plusmn004

D207plusmn022

d252plusmn026

A286plusmn030

b13

0plusmn014

B270plusmn028

c3

2175

6000

0lowastlowastlowast

trans-C

innamicacid

090plusmn008

C321plusmn034

c060plusmn005

D377plusmn036

a235plusmn025

A267plusmn024

d16

9plusmn017

B350plusmn036

b3

1060

0003lowastlowast

Total

980plusmn088

B34

96plusmn356

c72

5plusmn081

D45

57plusmn523

b5138plusmn436

A58

33plusmn621

a76

6plusmn080

C1588plusmn16

6dFlavanols

Catechin

026plusmn003

C093plusmn008

c050plusmn004

B314plusmn044

b457plusmn050

A519plusmn056

and

nd3

589952

000

0lowastlowastlowast

Epicatechin

067plusmn008

239plusmn024

ndnd

ndNd

ndnd

31713630

000

0lowastlowastlowast

Total

093plusmn008

B332plusmn035

b050plusmn006

C314plusmn029

c457plusmn052

A519plusmn062

and

ndFlavanon

esNaringenin

120plusmn011

C428plusmn051

b115plusmn012

D72

3plusmn077

a211plusmn022

A240plusmn025

d13

1plusmn014

B272plusmn028

c3

3614

2000

0lowastlowastlowast

Total

120plusmn010

C428plusmn045

b11

5plusmn010

D72

3plusmn077

a211plusmn023

A240plusmn026

d13

1plusmn012

B272plusmn028

c

Flavon

ols

Myristin

087plusmn009

C310plusmn033

d059plusmn006

D371plusmn036

c345plusmn041

A392plusmn041

b304plusmn040

B630plusmn074

a3

23583

000

0lowastlowastlowast

Quercetin

dihydrate

094plusmn008

C33

5plusmn035

a033plusmn004

D207plusmn022

b10

6plusmn010

B12

0plusmn013

c16

1plusmn017

A334plusmn036

a3

29686

000

0lowastlowastlowast

Camph

erol

057plusmn006

D203plusmn025

c12

6plusmn013

C79

2plusmn084

a13

5plusmn014

B15

3plusmn016

d15

5plusmn016

A321plusmn034

b3

4442309

000

0lowastlowastlowast

Total

238plusmn024

C849plusmn099

c218plusmn022

D1370plusmn14

5a586plusmn066

B665plusmn076

d620plusmn065

A1286plusmn14

2bFlavon

esAp

igenin

140plusmn013

C499plusmn052

b041plusmn005

D258plusmn026

d278plusmn029

B316plusmn041

c296plusmn033

A614plusmn072

a3

129479

6000

0lowastlowastlowast

Flavon

e545plusmn061

C1944plusmn236

b18

9plusmn017

D1188plusmn211

c601plusmn059

B682plusmn073

d1566plusmn17

2A3247plusmn423

a3

5334547

000

0lowastlowastlowast

Total

738plusmn088

C26

33plusmn277

b230plusmn022

D1446plusmn16

6c879plusmn099

B998plusmn12

3d1862plusmn201

A38

61plusmn

413

a

Cou

marins

Cou

marin

459plusmn051

B1638plusmn17

8b16

7plusmn017

C1050plusmn15

6d1334plusmn114A

1515plusmn17

9c1331plusmn15

1A2760plusmn286

a3

61887

000

0lowastlowastlowast

Total

459plusmn055

B1638plusmn18

8b16

7plusmn018

C1050plusmn12

3d1334plusmn16

2A1515plusmn16

2c1331plusmn14

3A2760plusmn297

a

Total

2803plusmn312

C100plusmn92

3a1591plusmn

165D

100plusmn846

a88

08plusmn759A

100plusmn98

7a48

23plusmn521

B100plusmn899

a

Thev

alueso

fthe


fpheno

liccompo

unds

representthe

averageo

fthree

replicates

(119899=3)n

dno

tdetectedLette

rs(andashd

(lower


ndAndash

D(capita

lletters)b

etween

amou

nts)indicatesig

nificantd

ifferencesa


ificant


offre

edom

119865Fish
















50corre-


50values










References






































































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of





2) was


6ndashC22








2SO4were














119860blank] times 100 (1)


50) was



3Fe (CN)

6) and incubated


50) was









a

bc

bc

0

02

04

06

08

1

12

14

16

18


Esse

ntia

l oil

yiel

d (

ww

)


































































Table 2 Continued


















minus10minus08

minus06minus04

minus04

minus02

minus02

00

025035

045055

065075

085095

00

02

02

04

04

06

06

08

Axe 1(18

48)

Axe 2 (1104)

Axe

3(692

) Jeacuterissa

Boukornine

Kairouan

Bou Salem




6ndashC3ndashC6)





a

a

a

b

b

bc

c

c

dd d

0

20

40

60

80

100

120

140

160

180



Bou Salem




minus02

00

02

04

06

08

Axe

3(446

)

Jeacuterissa

Boukornine

Kairouan

Bou Salem

025

040

055

070

085

100

Axe 1(24

45)

minus04minus02

0002

0406

0810

12

Axe 2 (951)






50of 612 plusmn 306 120583gmL



Table3Analysis

ofvaria

nce(ANOVA

)and

phenoliccompo

sitionin

(w

w)a

ndqu

antity(120583ggD

W)o

fmethano

lextractso

fthe

aeria

lpartsof

wormwoo

d(A

absinthium

L)

Com

poun

dCollectionregion

df119865

119875

BouSalem

Kairo

uan

Jeris

saBo

ukornine

120583ggD

W

120583ggD

W

120583ggD

W

120583ggD

W

Phenolicacids

Tann

icacid

037plusmn004

D13

2plusmn011

d073plusmn008

C459plusmn051

b334plusmn345

A3792plusmn402

a19

9plusmn201

B413plusmn044

c3

2028659

000

0lowastlowastlowast

Gallic

acid

038plusmn004

D13

6plusmn012

c041plusmn003

C258plusmn026

b475plusmn055

A539plusmn061

a063plusmn007

B13

1plusmn011

d3

2925813

000

0lowastlowastlowast

Chlorogenica

cidhemihydrate

336plusmn042

A1199plusmn13

2a14

9plusmn016

C93

7plusmn098

b233plusmn031

B265plusmn031

c081plusmn009

D16

8plusmn017

d3

898916

2000

0lowastlowastlowast

Caffeicacid

ndnd

152plusmn016

A95

5plusmn099

a14

3plusmn013

B16

2plusmn014

bnd

nd3

8639266

000

0lowastlowastlowast

Vanillica

cid

047plusmn005

B16

8plusmn015

b094plusmn008

A591plusmn066

and

Nd

ndnd

31867878

000

0lowastlowastlowast

Syrin

gica

cid

ndnd

ndnd

126plusmn014

143plusmn012

ndnd

3681633

000

0lowastlowastlowast

Ferulic

acid

035plusmn004

D12

5plusmn011

d068plusmn007

B427plusmn044

a117plusmn015

A13

3plusmn010

c065plusmn007

C13

5plusmn012

b3

417574

000

0lowastlowastlowast

p-Cou

maricacid

306plusmn041

A1092plusmn12

3a055plusmn006

D346plusmn041

b217plusmn023

B246plusmn025

c059plusmn006

C12

2plusmn011

d3

4768092

000

0lowastlowastlowast

Rosm

arinicacid

091plusmn008

C325plusmn041

a033plusmn004

D207plusmn022

d252plusmn026

A286plusmn030

b13

0plusmn014

B270plusmn028

c3

2175

6000

0lowastlowastlowast

trans-C

innamicacid

090plusmn008

C321plusmn034

c060plusmn005

D377plusmn036

a235plusmn025

A267plusmn024

d16

9plusmn017

B350plusmn036

b3

1060

0003lowastlowast

Total

980plusmn088

B34

96plusmn356

c72

5plusmn081

D45

57plusmn523

b5138plusmn436

A58

33plusmn621

a76

6plusmn080

C1588plusmn16

6dFlavanols

Catechin

026plusmn003

C093plusmn008

c050plusmn004

B314plusmn044

b457plusmn050

A519plusmn056

and

nd3

589952

000

0lowastlowastlowast

Epicatechin

067plusmn008

239plusmn024

ndnd

ndNd

ndnd

31713630

000

0lowastlowastlowast

Total

093plusmn008

B332plusmn035

b050plusmn006

C314plusmn029

c457plusmn052

A519plusmn062

and

ndFlavanon

esNaringenin

120plusmn011

C428plusmn051

b115plusmn012

D72

3plusmn077

a211plusmn022

A240plusmn025

d13

1plusmn014

B272plusmn028

c3

3614

2000

0lowastlowastlowast

Total

120plusmn010

C428plusmn045

b11

5plusmn010

D72

3plusmn077

a211plusmn023

A240plusmn026

d13

1plusmn012

B272plusmn028

c

Flavon

ols

Myristin

087plusmn009

C310plusmn033

d059plusmn006

D371plusmn036

c345plusmn041

A392plusmn041

b304plusmn040

B630plusmn074

a3

23583

000

0lowastlowastlowast

Quercetin

dihydrate

094plusmn008

C33

5plusmn035

a033plusmn004

D207plusmn022

b10

6plusmn010

B12

0plusmn013

c16

1plusmn017

A334plusmn036

a3

29686

000

0lowastlowastlowast

Camph

erol

057plusmn006

D203plusmn025

c12

6plusmn013

C79

2plusmn084

a13

5plusmn014

B15

3plusmn016

d15

5plusmn016

A321plusmn034

b3

4442309

000

0lowastlowastlowast

Total

238plusmn024

C849plusmn099

c218plusmn022

D1370plusmn14

5a586plusmn066

B665plusmn076

d620plusmn065

A1286plusmn14

2bFlavon

esAp

igenin

140plusmn013

C499plusmn052

b041plusmn005

D258plusmn026

d278plusmn029

B316plusmn041

c296plusmn033

A614plusmn072

a3

129479

6000

0lowastlowastlowast

Flavon

e545plusmn061

C1944plusmn236

b18

9plusmn017

D1188plusmn211

c601plusmn059

B682plusmn073

d1566plusmn17

2A3247plusmn423

a3

5334547

000

0lowastlowastlowast

Total

738plusmn088

C26

33plusmn277

b230plusmn022

D1446plusmn16

6c879plusmn099

B998plusmn12

3d1862plusmn201

A38

61plusmn

413

a

Cou

marins

Cou

marin

459plusmn051

B1638plusmn17

8b16

7plusmn017

C1050plusmn15

6d1334plusmn114A

1515plusmn17

9c1331plusmn15

1A2760plusmn286

a3

61887

000

0lowastlowastlowast

Total

459plusmn055

B1638plusmn18

8b16

7plusmn018

C1050plusmn12

3d1334plusmn16

2A1515plusmn16

2c1331plusmn14

3A2760plusmn297

a

Total

2803plusmn312

C100plusmn92

3a1591plusmn

165D

100plusmn846

a88

08plusmn759A

100plusmn98

7a48

23plusmn521

B100plusmn899

a

Thev

alueso

fthe


fpheno

liccompo

unds

representthe

averageo

fthree

replicates

(119899=3)n

dno

tdetectedLette

rs(andashd

(lower


ndAndash

D(capita

lletters)b

etween

amou

nts)indicatesig

nificantd

ifferencesa


ificant


offre

edom

119865Fish
















50corre-


50values










References






































































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of






119860blank] times 100 (1)


50) was



3Fe (CN)

6) and incubated


50) was









a

bc

bc

0

02

04

06

08

1

12

14

16

18


Esse

ntia

l oil

yiel

d (

ww

)


































































Table 2 Continued


















minus10minus08

minus06minus04

minus04

minus02

minus02

00

025035

045055

065075

085095

00

02

02

04

04

06

06

08

Axe 1(18

48)

Axe 2 (1104)

Axe

3(692

) Jeacuterissa

Boukornine

Kairouan

Bou Salem




6ndashC3ndashC6)





a

a

a

b

b

bc

c

c

dd d

0

20

40

60

80

100

120

140

160

180



Bou Salem




minus02

00

02

04

06

08

Axe

3(446

)

Jeacuterissa

Boukornine

Kairouan

Bou Salem

025

040

055

070

085

100

Axe 1(24

45)

minus04minus02

0002

0406

0810

12

Axe 2 (951)






50of 612 plusmn 306 120583gmL



Table3Analysis

ofvaria

nce(ANOVA

)and

phenoliccompo

sitionin

(w

w)a

ndqu

antity(120583ggD

W)o

fmethano

lextractso

fthe

aeria

lpartsof

wormwoo

d(A

absinthium

L)

Com

poun

dCollectionregion

df119865

119875

BouSalem

Kairo

uan

Jeris

saBo

ukornine

120583ggD

W

120583ggD

W

120583ggD

W

120583ggD

W

Phenolicacids

Tann

icacid

037plusmn004

D13

2plusmn011

d073plusmn008

C459plusmn051

b334plusmn345

A3792plusmn402

a19

9plusmn201

B413plusmn044

c3

2028659

000

0lowastlowastlowast

Gallic

acid

038plusmn004

D13

6plusmn012

c041plusmn003

C258plusmn026

b475plusmn055

A539plusmn061

a063plusmn007

B13

1plusmn011

d3

2925813

000

0lowastlowastlowast

Chlorogenica

cidhemihydrate

336plusmn042

A1199plusmn13

2a14

9plusmn016

C93

7plusmn098

b233plusmn031

B265plusmn031

c081plusmn009

D16

8plusmn017

d3

898916

2000

0lowastlowastlowast

Caffeicacid

ndnd

152plusmn016

A95

5plusmn099

a14

3plusmn013

B16

2plusmn014

bnd

nd3

8639266

000

0lowastlowastlowast

Vanillica

cid

047plusmn005

B16

8plusmn015

b094plusmn008

A591plusmn066

and

Nd

ndnd

31867878

000

0lowastlowastlowast

Syrin

gica

cid

ndnd

ndnd

126plusmn014

143plusmn012

ndnd

3681633

000

0lowastlowastlowast

Ferulic

acid

035plusmn004

D12

5plusmn011

d068plusmn007

B427plusmn044

a117plusmn015

A13

3plusmn010

c065plusmn007

C13

5plusmn012

b3

417574

000

0lowastlowastlowast

p-Cou

maricacid

306plusmn041

A1092plusmn12

3a055plusmn006

D346plusmn041

b217plusmn023

B246plusmn025

c059plusmn006

C12

2plusmn011

d3

4768092

000

0lowastlowastlowast

Rosm

arinicacid

091plusmn008

C325plusmn041

a033plusmn004

D207plusmn022

d252plusmn026

A286plusmn030

b13

0plusmn014

B270plusmn028

c3

2175

6000

0lowastlowastlowast

trans-C

innamicacid

090plusmn008

C321plusmn034

c060plusmn005

D377plusmn036

a235plusmn025

A267plusmn024

d16

9plusmn017

B350plusmn036

b3

1060

0003lowastlowast

Total

980plusmn088

B34

96plusmn356

c72

5plusmn081

D45

57plusmn523

b5138plusmn436

A58

33plusmn621

a76

6plusmn080

C1588plusmn16

6dFlavanols

Catechin

026plusmn003

C093plusmn008

c050plusmn004

B314plusmn044

b457plusmn050

A519plusmn056

and

nd3

589952

000

0lowastlowastlowast

Epicatechin

067plusmn008

239plusmn024

ndnd

ndNd

ndnd

31713630

000

0lowastlowastlowast

Total

093plusmn008

B332plusmn035

b050plusmn006

C314plusmn029

c457plusmn052

A519plusmn062

and

ndFlavanon

esNaringenin

120plusmn011

C428plusmn051

b115plusmn012

D72

3plusmn077

a211plusmn022

A240plusmn025

d13

1plusmn014

B272plusmn028

c3

3614

2000

0lowastlowastlowast

Total

120plusmn010

C428plusmn045

b11

5plusmn010

D72

3plusmn077

a211plusmn023

A240plusmn026

d13

1plusmn012

B272plusmn028

c

Flavon

ols

Myristin

087plusmn009

C310plusmn033

d059plusmn006

D371plusmn036

c345plusmn041

A392plusmn041

b304plusmn040

B630plusmn074

a3

23583

000

0lowastlowastlowast

Quercetin

dihydrate

094plusmn008

C33

5plusmn035

a033plusmn004

D207plusmn022

b10

6plusmn010

B12

0plusmn013

c16

1plusmn017

A334plusmn036

a3

29686

000

0lowastlowastlowast

Camph

erol

057plusmn006

D203plusmn025

c12

6plusmn013

C79

2plusmn084

a13

5plusmn014

B15

3plusmn016

d15

5plusmn016

A321plusmn034

b3

4442309

000

0lowastlowastlowast

Total

238plusmn024

C849plusmn099

c218plusmn022

D1370plusmn14

5a586plusmn066

B665plusmn076

d620plusmn065

A1286plusmn14

2bFlavon

esAp

igenin

140plusmn013

C499plusmn052

b041plusmn005

D258plusmn026

d278plusmn029

B316plusmn041

c296plusmn033

A614plusmn072

a3

129479

6000

0lowastlowastlowast

Flavon

e545plusmn061

C1944plusmn236

b18

9plusmn017

D1188plusmn211

c601plusmn059

B682plusmn073

d1566plusmn17

2A3247plusmn423

a3

5334547

000

0lowastlowastlowast

Total

738plusmn088

C26

33plusmn277

b230plusmn022

D1446plusmn16

6c879plusmn099

B998plusmn12

3d1862plusmn201

A38

61plusmn

413

a

Cou

marins

Cou

marin

459plusmn051

B1638plusmn17

8b16

7plusmn017

C1050plusmn15

6d1334plusmn114A

1515plusmn17

9c1331plusmn15

1A2760plusmn286

a3

61887

000

0lowastlowastlowast

Total

459plusmn055

B1638plusmn18

8b16

7plusmn018

C1050plusmn12

3d1334plusmn16

2A1515plusmn16

2c1331plusmn14

3A2760plusmn297

a

Total

2803plusmn312

C100plusmn92

3a1591plusmn

165D

100plusmn846

a88

08plusmn759A

100plusmn98

7a48

23plusmn521

B100plusmn899

a

Thev

alueso

fthe


fpheno

liccompo

unds

representthe

averageo

fthree

replicates

(119899=3)n

dno

tdetectedLette

rs(andashd

(lower


ndAndash

D(capita

lletters)b

etween

amou

nts)indicatesig

nificantd

ifferencesa


ificant


offre

edom

119865Fish
















50corre-


50values










References






































































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


a

bc

bc

0

02

04

06

08

1

12

14

16

18


Esse

ntia

l oil

yiel

d (

ww

)


































































Table 2 Continued


















minus10minus08

minus06minus04

minus04

minus02

minus02

00

025035

045055

065075

085095

00

02

02

04

04

06

06

08

Axe 1(18

48)

Axe 2 (1104)

Axe

3(692

) Jeacuterissa

Boukornine

Kairouan

Bou Salem




6ndashC3ndashC6)





a

a

a

b

b

bc

c

c

dd d

0

20

40

60

80

100

120

140

160

180



Bou Salem




minus02

00

02

04

06

08

Axe

3(446

)

Jeacuterissa

Boukornine

Kairouan

Bou Salem

025

040

055

070

085

100

Axe 1(24

45)

minus04minus02

0002

0406

0810

12

Axe 2 (951)






50of 612 plusmn 306 120583gmL



Table3Analysis

ofvaria

nce(ANOVA

)and

phenoliccompo

sitionin

(w

w)a

ndqu

antity(120583ggD

W)o

fmethano

lextractso

fthe

aeria

lpartsof

wormwoo

d(A

absinthium

L)

Com

poun

dCollectionregion

df119865

119875

BouSalem

Kairo

uan

Jeris

saBo

ukornine

120583ggD

W

120583ggD

W

120583ggD

W

120583ggD

W

Phenolicacids

Tann

icacid

037plusmn004

D13

2plusmn011

d073plusmn008

C459plusmn051

b334plusmn345

A3792plusmn402

a19

9plusmn201

B413plusmn044

c3

2028659

000

0lowastlowastlowast

Gallic

acid

038plusmn004

D13

6plusmn012

c041plusmn003

C258plusmn026

b475plusmn055

A539plusmn061

a063plusmn007

B13

1plusmn011

d3

2925813

000

0lowastlowastlowast

Chlorogenica

cidhemihydrate

336plusmn042

A1199plusmn13

2a14

9plusmn016

C93

7plusmn098

b233plusmn031

B265plusmn031

c081plusmn009

D16

8plusmn017

d3

898916

2000

0lowastlowastlowast

Caffeicacid

ndnd

152plusmn016

A95

5plusmn099

a14

3plusmn013

B16

2plusmn014

bnd

nd3

8639266

000

0lowastlowastlowast

Vanillica

cid

047plusmn005

B16

8plusmn015

b094plusmn008

A591plusmn066

and

Nd

ndnd

31867878

000

0lowastlowastlowast

Syrin

gica

cid

ndnd

ndnd

126plusmn014

143plusmn012

ndnd

3681633

000

0lowastlowastlowast

Ferulic

acid

035plusmn004

D12

5plusmn011

d068plusmn007

B427plusmn044

a117plusmn015

A13

3plusmn010

c065plusmn007

C13

5plusmn012

b3

417574

000

0lowastlowastlowast

p-Cou

maricacid

306plusmn041

A1092plusmn12

3a055plusmn006

D346plusmn041

b217plusmn023

B246plusmn025

c059plusmn006

C12

2plusmn011

d3

4768092

000

0lowastlowastlowast

Rosm

arinicacid

091plusmn008

C325plusmn041

a033plusmn004

D207plusmn022

d252plusmn026

A286plusmn030

b13

0plusmn014

B270plusmn028

c3

2175

6000

0lowastlowastlowast

trans-C

innamicacid

090plusmn008

C321plusmn034

c060plusmn005

D377plusmn036

a235plusmn025

A267plusmn024

d16

9plusmn017

B350plusmn036

b3

1060

0003lowastlowast

Total

980plusmn088

B34

96plusmn356

c72

5plusmn081

D45

57plusmn523

b5138plusmn436

A58

33plusmn621

a76

6plusmn080

C1588plusmn16

6dFlavanols

Catechin

026plusmn003

C093plusmn008

c050plusmn004

B314plusmn044

b457plusmn050

A519plusmn056

and

nd3

589952

000

0lowastlowastlowast

Epicatechin

067plusmn008

239plusmn024

ndnd

ndNd

ndnd

31713630

000

0lowastlowastlowast

Total

093plusmn008

B332plusmn035

b050plusmn006

C314plusmn029

c457plusmn052

A519plusmn062

and

ndFlavanon

esNaringenin

120plusmn011

C428plusmn051

b115plusmn012

D72

3plusmn077

a211plusmn022

A240plusmn025

d13

1plusmn014

B272plusmn028

c3

3614

2000

0lowastlowastlowast

Total

120plusmn010

C428plusmn045

b11

5plusmn010

D72

3plusmn077

a211plusmn023

A240plusmn026

d13

1plusmn012

B272plusmn028

c

Flavon

ols

Myristin

087plusmn009

C310plusmn033

d059plusmn006

D371plusmn036

c345plusmn041

A392plusmn041

b304plusmn040

B630plusmn074

a3

23583

000

0lowastlowastlowast

Quercetin

dihydrate

094plusmn008

C33

5plusmn035

a033plusmn004

D207plusmn022

b10

6plusmn010

B12

0plusmn013

c16

1plusmn017

A334plusmn036

a3

29686

000

0lowastlowastlowast

Camph

erol

057plusmn006

D203plusmn025

c12

6plusmn013

C79

2plusmn084

a13

5plusmn014

B15

3plusmn016

d15

5plusmn016

A321plusmn034

b3

4442309

000

0lowastlowastlowast

Total

238plusmn024

C849plusmn099

c218plusmn022

D1370plusmn14

5a586plusmn066

B665plusmn076

d620plusmn065

A1286plusmn14

2bFlavon

esAp

igenin

140plusmn013

C499plusmn052

b041plusmn005

D258plusmn026

d278plusmn029

B316plusmn041

c296plusmn033

A614plusmn072

a3

129479

6000

0lowastlowastlowast

Flavon

e545plusmn061

C1944plusmn236

b18

9plusmn017

D1188plusmn211

c601plusmn059

B682plusmn073

d1566plusmn17

2A3247plusmn423

a3

5334547

000

0lowastlowastlowast

Total

738plusmn088

C26

33plusmn277

b230plusmn022

D1446plusmn16

6c879plusmn099

B998plusmn12

3d1862plusmn201

A38

61plusmn

413

a

Cou

marins

Cou

marin

459plusmn051

B1638plusmn17

8b16

7plusmn017

C1050plusmn15

6d1334plusmn114A

1515plusmn17

9c1331plusmn15

1A2760plusmn286

a3

61887

000

0lowastlowastlowast

Total

459plusmn055

B1638plusmn18

8b16

7plusmn018

C1050plusmn12

3d1334plusmn16

2A1515plusmn16

2c1331plusmn14

3A2760plusmn297

a

Total

2803plusmn312

C100plusmn92

3a1591plusmn

165D

100plusmn846

a88

08plusmn759A

100plusmn98

7a48

23plusmn521

B100plusmn899

a

Thev

alueso

fthe


fpheno

liccompo

unds

representthe

averageo

fthree

replicates

(119899=3)n

dno

tdetectedLette

rs(andashd

(lower


ndAndash

D(capita

lletters)b

etween

amou

nts)indicatesig

nificantd

ifferencesa


ificant


offre

edom

119865Fish
















50corre-


50values










References






































































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of























































Table 2 Continued


















minus10minus08

minus06minus04

minus04

minus02

minus02

00

025035

045055

065075

085095

00

02

02

04

04

06

06

08

Axe 1(18

48)

Axe 2 (1104)

Axe

3(692

) Jeacuterissa

Boukornine

Kairouan

Bou Salem




6ndashC3ndashC6)





a

a

a

b

b

bc

c

c

dd d

0

20

40

60

80

100

120

140

160

180



Bou Salem




minus02

00

02

04

06

08

Axe

3(446

)

Jeacuterissa

Boukornine

Kairouan

Bou Salem

025

040

055

070

085

100

Axe 1(24

45)

minus04minus02

0002

0406

0810

12

Axe 2 (951)






50of 612 plusmn 306 120583gmL



Table3Analysis

ofvaria

nce(ANOVA

)and

phenoliccompo

sitionin

(w

w)a

ndqu

antity(120583ggD

W)o

fmethano

lextractso

fthe

aeria

lpartsof

wormwoo

d(A

absinthium

L)

Com

poun

dCollectionregion

df119865

119875

BouSalem

Kairo

uan

Jeris

saBo

ukornine

120583ggD

W

120583ggD

W

120583ggD

W

120583ggD

W

Phenolicacids

Tann

icacid

037plusmn004

D13

2plusmn011

d073plusmn008

C459plusmn051

b334plusmn345

A3792plusmn402

a19

9plusmn201

B413plusmn044

c3

2028659

000

0lowastlowastlowast

Gallic

acid

038plusmn004

D13

6plusmn012

c041plusmn003

C258plusmn026

b475plusmn055

A539plusmn061

a063plusmn007

B13

1plusmn011

d3

2925813

000

0lowastlowastlowast

Chlorogenica

cidhemihydrate

336plusmn042

A1199plusmn13

2a14

9plusmn016

C93

7plusmn098

b233plusmn031

B265plusmn031

c081plusmn009

D16

8plusmn017

d3

898916

2000

0lowastlowastlowast

Caffeicacid

ndnd

152plusmn016

A95

5plusmn099

a14

3plusmn013

B16

2plusmn014

bnd

nd3

8639266

000

0lowastlowastlowast

Vanillica

cid

047plusmn005

B16

8plusmn015

b094plusmn008

A591plusmn066

and

Nd

ndnd

31867878

000

0lowastlowastlowast

Syrin

gica

cid

ndnd

ndnd

126plusmn014

143plusmn012

ndnd

3681633

000

0lowastlowastlowast

Ferulic

acid

035plusmn004

D12

5plusmn011

d068plusmn007

B427plusmn044

a117plusmn015

A13

3plusmn010

c065plusmn007

C13

5plusmn012

b3

417574

000

0lowastlowastlowast

p-Cou

maricacid

306plusmn041

A1092plusmn12

3a055plusmn006

D346plusmn041

b217plusmn023

B246plusmn025

c059plusmn006

C12

2plusmn011

d3

4768092

000

0lowastlowastlowast

Rosm

arinicacid

091plusmn008

C325plusmn041

a033plusmn004

D207plusmn022

d252plusmn026

A286plusmn030

b13

0plusmn014

B270plusmn028

c3

2175

6000

0lowastlowastlowast

trans-C

innamicacid

090plusmn008

C321plusmn034

c060plusmn005

D377plusmn036

a235plusmn025

A267plusmn024

d16

9plusmn017

B350plusmn036

b3

1060

0003lowastlowast

Total

980plusmn088

B34

96plusmn356

c72

5plusmn081

D45

57plusmn523

b5138plusmn436

A58

33plusmn621

a76

6plusmn080

C1588plusmn16

6dFlavanols

Catechin

026plusmn003

C093plusmn008

c050plusmn004

B314plusmn044

b457plusmn050

A519plusmn056

and

nd3

589952

000

0lowastlowastlowast

Epicatechin

067plusmn008

239plusmn024

ndnd

ndNd

ndnd

31713630

000

0lowastlowastlowast

Total

093plusmn008

B332plusmn035

b050plusmn006

C314plusmn029

c457plusmn052

A519plusmn062

and

ndFlavanon

esNaringenin

120plusmn011

C428plusmn051

b115plusmn012

D72

3plusmn077

a211plusmn022

A240plusmn025

d13

1plusmn014

B272plusmn028

c3

3614

2000

0lowastlowastlowast

Total

120plusmn010

C428plusmn045

b11

5plusmn010

D72

3plusmn077

a211plusmn023

A240plusmn026

d13

1plusmn012

B272plusmn028

c

Flavon

ols

Myristin

087plusmn009

C310plusmn033

d059plusmn006

D371plusmn036

c345plusmn041

A392plusmn041

b304plusmn040

B630plusmn074

a3

23583

000

0lowastlowastlowast

Quercetin

dihydrate

094plusmn008

C33

5plusmn035

a033plusmn004

D207plusmn022

b10

6plusmn010

B12

0plusmn013

c16

1plusmn017

A334plusmn036

a3

29686

000

0lowastlowastlowast

Camph

erol

057plusmn006

D203plusmn025

c12

6plusmn013

C79

2plusmn084

a13

5plusmn014

B15

3plusmn016

d15

5plusmn016

A321plusmn034

b3

4442309

000

0lowastlowastlowast

Total

238plusmn024

C849plusmn099

c218plusmn022

D1370plusmn14

5a586plusmn066

B665plusmn076

d620plusmn065

A1286plusmn14

2bFlavon

esAp

igenin

140plusmn013

C499plusmn052

b041plusmn005

D258plusmn026

d278plusmn029

B316plusmn041

c296plusmn033

A614plusmn072

a3

129479

6000

0lowastlowastlowast

Flavon

e545plusmn061

C1944plusmn236

b18

9plusmn017

D1188plusmn211

c601plusmn059

B682plusmn073

d1566plusmn17

2A3247plusmn423

a3

5334547

000

0lowastlowastlowast

Total

738plusmn088

C26

33plusmn277

b230plusmn022

D1446plusmn16

6c879plusmn099

B998plusmn12

3d1862plusmn201

A38

61plusmn

413

a

Cou

marins

Cou

marin

459plusmn051

B1638plusmn17

8b16

7plusmn017

C1050plusmn15

6d1334plusmn114A

1515plusmn17

9c1331plusmn15

1A2760plusmn286

a3

61887

000

0lowastlowastlowast

Total

459plusmn055

B1638plusmn18

8b16

7plusmn018

C1050plusmn12

3d1334plusmn16

2A1515plusmn16

2c1331plusmn14

3A2760plusmn297

a

Total

2803plusmn312

C100plusmn92

3a1591plusmn

165D

100plusmn846

a88

08plusmn759A

100plusmn98

7a48

23plusmn521

B100plusmn899

a

Thev

alueso

fthe


fpheno

liccompo

unds

representthe

averageo

fthree

replicates

(119899=3)n

dno

tdetectedLette

rs(andashd

(lower


ndAndash

D(capita

lletters)b

etween

amou

nts)indicatesig

nificantd

ifferencesa


ificant


offre

edom

119865Fish
















50corre-


50values










References






































































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


a

a

a

b

b

bc

c

c

dd d

0

20

40

60

80

100

120

140

160

180



Bou Salem




minus02

00

02

04

06

08

Axe

3(446

)

Jeacuterissa

Boukornine

Kairouan

Bou Salem

025

040

055

070

085

100

Axe 1(24

45)

minus04minus02

0002

0406

0810

12

Axe 2 (951)






50of 612 plusmn 306 120583gmL



Table3Analysis

ofvaria

nce(ANOVA

)and

phenoliccompo

sitionin

(w

w)a

ndqu

antity(120583ggD

W)o

fmethano

lextractso

fthe

aeria

lpartsof

wormwoo

d(A

absinthium

L)

Com

poun

dCollectionregion

df119865

119875

BouSalem

Kairo

uan

Jeris

saBo

ukornine

120583ggD

W

120583ggD

W

120583ggD

W

120583ggD

W

Phenolicacids

Tann

icacid

037plusmn004

D13

2plusmn011

d073plusmn008

C459plusmn051

b334plusmn345

A3792plusmn402

a19

9plusmn201

B413plusmn044

c3

2028659

000

0lowastlowastlowast

Gallic

acid

038plusmn004

D13

6plusmn012

c041plusmn003

C258plusmn026

b475plusmn055

A539plusmn061

a063plusmn007

B13

1plusmn011

d3

2925813

000

0lowastlowastlowast

Chlorogenica

cidhemihydrate

336plusmn042

A1199plusmn13

2a14

9plusmn016

C93

7plusmn098

b233plusmn031

B265plusmn031

c081plusmn009

D16

8plusmn017

d3

898916

2000

0lowastlowastlowast

Caffeicacid

ndnd

152plusmn016

A95

5plusmn099

a14

3plusmn013

B16

2plusmn014

bnd

nd3

8639266

000

0lowastlowastlowast

Vanillica

cid

047plusmn005

B16

8plusmn015

b094plusmn008

A591plusmn066

and

Nd

ndnd

31867878

000

0lowastlowastlowast

Syrin

gica

cid

ndnd

ndnd

126plusmn014

143plusmn012

ndnd

3681633

000

0lowastlowastlowast

Ferulic

acid

035plusmn004

D12

5plusmn011

d068plusmn007

B427plusmn044

a117plusmn015

A13

3plusmn010

c065plusmn007

C13

5plusmn012

b3

417574

000

0lowastlowastlowast

p-Cou

maricacid

306plusmn041

A1092plusmn12

3a055plusmn006

D346plusmn041

b217plusmn023

B246plusmn025

c059plusmn006

C12

2plusmn011

d3

4768092

000

0lowastlowastlowast

Rosm

arinicacid

091plusmn008

C325plusmn041

a033plusmn004

D207plusmn022

d252plusmn026

A286plusmn030

b13

0plusmn014

B270plusmn028

c3

2175

6000

0lowastlowastlowast

trans-C

innamicacid

090plusmn008

C321plusmn034

c060plusmn005

D377plusmn036

a235plusmn025

A267plusmn024

d16

9plusmn017

B350plusmn036

b3

1060

0003lowastlowast

Total

980plusmn088

B34

96plusmn356

c72

5plusmn081

D45

57plusmn523

b5138plusmn436

A58

33plusmn621

a76

6plusmn080

C1588plusmn16

6dFlavanols

Catechin

026plusmn003

C093plusmn008

c050plusmn004

B314plusmn044

b457plusmn050

A519plusmn056

and

nd3

589952

000

0lowastlowastlowast

Epicatechin

067plusmn008

239plusmn024

ndnd

ndNd

ndnd

31713630

000

0lowastlowastlowast

Total

093plusmn008

B332plusmn035

b050plusmn006

C314plusmn029

c457plusmn052

A519plusmn062

and

ndFlavanon

esNaringenin

120plusmn011

C428plusmn051

b115plusmn012

D72

3plusmn077

a211plusmn022

A240plusmn025

d13

1plusmn014

B272plusmn028

c3

3614

2000

0lowastlowastlowast

Total

120plusmn010

C428plusmn045

b11

5plusmn010

D72

3plusmn077

a211plusmn023

A240plusmn026

d13

1plusmn012

B272plusmn028

c

Flavon

ols

Myristin

087plusmn009

C310plusmn033

d059plusmn006

D371plusmn036

c345plusmn041

A392plusmn041

b304plusmn040

B630plusmn074

a3

23583

000

0lowastlowastlowast

Quercetin

dihydrate

094plusmn008

C33

5plusmn035

a033plusmn004

D207plusmn022

b10

6plusmn010

B12

0plusmn013

c16

1plusmn017

A334plusmn036

a3

29686

000

0lowastlowastlowast

Camph

erol

057plusmn006

D203plusmn025

c12

6plusmn013

C79

2plusmn084

a13

5plusmn014

B15

3plusmn016

d15

5plusmn016

A321plusmn034

b3

4442309

000

0lowastlowastlowast

Total

238plusmn024

C849plusmn099

c218plusmn022

D1370plusmn14

5a586plusmn066

B665plusmn076

d620plusmn065

A1286plusmn14

2bFlavon

esAp

igenin

140plusmn013

C499plusmn052

b041plusmn005

D258plusmn026

d278plusmn029

B316plusmn041

c296plusmn033

A614plusmn072

a3

129479

6000

0lowastlowastlowast

Flavon

e545plusmn061

C1944plusmn236

b18

9plusmn017

D1188plusmn211

c601plusmn059

B682plusmn073

d1566plusmn17

2A3247plusmn423

a3

5334547

000

0lowastlowastlowast

Total

738plusmn088

C26

33plusmn277

b230plusmn022

D1446plusmn16

6c879plusmn099

B998plusmn12

3d1862plusmn201

A38

61plusmn

413

a

Cou

marins

Cou

marin

459plusmn051

B1638plusmn17

8b16

7plusmn017

C1050plusmn15

6d1334plusmn114A

1515plusmn17

9c1331plusmn15

1A2760plusmn286

a3

61887

000

0lowastlowastlowast

Total

459plusmn055

B1638plusmn18

8b16

7plusmn018

C1050plusmn12

3d1334plusmn16

2A1515plusmn16

2c1331plusmn14

3A2760plusmn297

a

Total

2803plusmn312

C100plusmn92

3a1591plusmn

165D

100plusmn846

a88

08plusmn759A

100plusmn98

7a48

23plusmn521

B100plusmn899

a

Thev

alueso

fthe


fpheno

liccompo

unds

representthe

averageo

fthree

replicates

(119899=3)n

dno

tdetectedLette

rs(andashd

(lower


ndAndash

D(capita

lletters)b

etween

amou

nts)indicatesig

nificantd

ifferencesa


ificant


offre

edom

119865Fish
















50corre-


50values










References






































































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


Table3Analysis

ofvaria

nce(ANOVA

)and

phenoliccompo

sitionin

(w

w)a

ndqu

antity(120583ggD

W)o

fmethano

lextractso

fthe

aeria

lpartsof

wormwoo

d(A

absinthium

L)

Com

poun

dCollectionregion

df119865

119875

BouSalem

Kairo

uan

Jeris

saBo

ukornine

120583ggD

W

120583ggD

W

120583ggD

W

120583ggD

W

Phenolicacids

Tann

icacid

037plusmn004

D13

2plusmn011

d073plusmn008

C459plusmn051

b334plusmn345

A3792plusmn402

a19

9plusmn201

B413plusmn044

c3

2028659

000

0lowastlowastlowast

Gallic

acid

038plusmn004

D13

6plusmn012

c041plusmn003

C258plusmn026

b475plusmn055

A539plusmn061

a063plusmn007

B13

1plusmn011

d3

2925813

000

0lowastlowastlowast

Chlorogenica

cidhemihydrate

336plusmn042

A1199plusmn13

2a14

9plusmn016

C93

7plusmn098

b233plusmn031

B265plusmn031

c081plusmn009

D16

8plusmn017

d3

898916

2000

0lowastlowastlowast

Caffeicacid

ndnd

152plusmn016

A95

5plusmn099

a14

3plusmn013

B16

2plusmn014

bnd

nd3

8639266

000

0lowastlowastlowast

Vanillica

cid

047plusmn005

B16

8plusmn015

b094plusmn008

A591plusmn066

and

Nd

ndnd

31867878

000

0lowastlowastlowast

Syrin

gica

cid

ndnd

ndnd

126plusmn014

143plusmn012

ndnd

3681633

000

0lowastlowastlowast

Ferulic

acid

035plusmn004

D12

5plusmn011

d068plusmn007

B427plusmn044

a117plusmn015

A13

3plusmn010

c065plusmn007

C13

5plusmn012

b3

417574

000

0lowastlowastlowast

p-Cou

maricacid

306plusmn041

A1092plusmn12

3a055plusmn006

D346plusmn041

b217plusmn023

B246plusmn025

c059plusmn006

C12

2plusmn011

d3

4768092

000

0lowastlowastlowast

Rosm

arinicacid

091plusmn008

C325plusmn041

a033plusmn004

D207plusmn022

d252plusmn026

A286plusmn030

b13

0plusmn014

B270plusmn028

c3

2175

6000

0lowastlowastlowast

trans-C

innamicacid

090plusmn008

C321plusmn034

c060plusmn005

D377plusmn036

a235plusmn025

A267plusmn024

d16

9plusmn017

B350plusmn036

b3

1060

0003lowastlowast

Total

980plusmn088

B34

96plusmn356

c72

5plusmn081

D45

57plusmn523

b5138plusmn436

A58

33plusmn621

a76

6plusmn080

C1588plusmn16

6dFlavanols

Catechin

026plusmn003

C093plusmn008

c050plusmn004

B314plusmn044

b457plusmn050

A519plusmn056

and

nd3

589952

000

0lowastlowastlowast

Epicatechin

067plusmn008

239plusmn024

ndnd

ndNd

ndnd

31713630

000

0lowastlowastlowast

Total

093plusmn008

B332plusmn035

b050plusmn006

C314plusmn029

c457plusmn052

A519plusmn062

and

ndFlavanon

esNaringenin

120plusmn011

C428plusmn051

b115plusmn012

D72

3plusmn077

a211plusmn022

A240plusmn025

d13

1plusmn014

B272plusmn028

c3

3614

2000

0lowastlowastlowast

Total

120plusmn010

C428plusmn045

b11

5plusmn010

D72

3plusmn077

a211plusmn023

A240plusmn026

d13

1plusmn012

B272plusmn028

c

Flavon

ols

Myristin

087plusmn009

C310plusmn033

d059plusmn006

D371plusmn036

c345plusmn041

A392plusmn041

b304plusmn040

B630plusmn074

a3

23583

000

0lowastlowastlowast

Quercetin

dihydrate

094plusmn008

C33

5plusmn035

a033plusmn004

D207plusmn022

b10

6plusmn010

B12

0plusmn013

c16

1plusmn017

A334plusmn036

a3

29686

000

0lowastlowastlowast

Camph

erol

057plusmn006

D203plusmn025

c12

6plusmn013

C79

2plusmn084

a13

5plusmn014

B15

3plusmn016

d15

5plusmn016

A321plusmn034

b3

4442309

000

0lowastlowastlowast

Total

238plusmn024

C849plusmn099

c218plusmn022

D1370plusmn14

5a586plusmn066

B665plusmn076

d620plusmn065

A1286plusmn14

2bFlavon

esAp

igenin

140plusmn013

C499plusmn052

b041plusmn005

D258plusmn026

d278plusmn029

B316plusmn041

c296plusmn033

A614plusmn072

a3

129479

6000

0lowastlowastlowast

Flavon

e545plusmn061

C1944plusmn236

b18

9plusmn017

D1188plusmn211

c601plusmn059

B682plusmn073

d1566plusmn17

2A3247plusmn423

a3

5334547

000

0lowastlowastlowast

Total

738plusmn088

C26

33plusmn277

b230plusmn022

D1446plusmn16

6c879plusmn099

B998plusmn12

3d1862plusmn201

A38

61plusmn

413

a

Cou

marins

Cou

marin

459plusmn051

B1638plusmn17

8b16

7plusmn017

C1050plusmn15

6d1334plusmn114A

1515plusmn17

9c1331plusmn15

1A2760plusmn286

a3

61887

000

0lowastlowastlowast

Total

459plusmn055

B1638plusmn18

8b16

7plusmn018

C1050plusmn12

3d1334plusmn16

2A1515plusmn16

2c1331plusmn14

3A2760plusmn297

a

Total

2803plusmn312

C100plusmn92

3a1591plusmn

165D

100plusmn846

a88

08plusmn759A

100plusmn98

7a48

23plusmn521

B100plusmn899

a

Thev

alueso

fthe


fpheno

liccompo

unds

representthe

averageo

fthree

replicates

(119899=3)n

dno

tdetectedLette

rs(andashd

(lower


ndAndash

D(capita

lletters)b

etween

amou

nts)indicatesig

nificantd

ifferencesa


ificant


offre

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Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of





References






































































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of











































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of

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