antioxidant activity ofessential oils from

a

ianizantisqerilshyddiem. Rse

species implicated in the plant network that contributes to sand were efcient against Listeria monocytogenes and Echerichia coli.[25]

Research Article

Accepted: 24 February 2013 Published online in Wiley Online Librarydune edication and restoration.[5,6] E. maritimum has also beenreported to exhibit different therapeutic uses in folk medicine.[7]

Several studies concerning the chemical compositions ofsolvent extracts from the Eryngium genus report triterpenesaponines,[8,9] acetylenic compounds[10] and polyphenols[11] as themain constituents. Essential oils of more than 20 Eryngium specieshave already been studied and their chemical compositionsare generally dominated by hydrocarbon sesquiterpenes suchas germacrene D,[12,13] bicyclogermacrene,[14] g-muurolene[14] andtrans-caryophyllene[15,16] or by non-terpenic oxygenated compoundssuch as trimethylbenzaldehyde and (E)-2-dodecenal.[1722]

To our knowledge, only two poster communications deal withthe chemical composition of E. maritimum essential oils. Both

The discovery of new natural antioxidants has been a chemicalchallenge in the last decade. Furthermore, toxic and/or mutageniceffects of many synthetic antioxidant components havesuggested plant antioxidants as an interesting alternative. It iscommonly assumed that almost all phenols can function as

* Correspondence to: Alain Muselli, Universit de Corse, UMR-CNRS 6134,Laboratoire Chimie des Produits Naturels, Campus Grimaldi, BP 52, 20250Corte, France. E-mail: muselli@univ-corse.fr

a Universit de Corse, UMR CNRS 6134 SPE, Laboratoire Chimie des Produitsessential oil, were tested for antioxidant properties using the DPPH and ABTS radical-scavenging activity tests. No meaningfulactivity could be attributed to sesquiterpene aldehydes and the corresponding alcohols but the total essential oil and theoxygenated fraction of E.maritimum both demonstrated strong antioxidant properties. Copyright 2013 JohnWiley & Sons, Ltd.

Keywords: Eryngium maritimum L.; essential oil; antioxidant activities; chemical variability; aldehyde sesquiterpenes

IntroductionThe genus Eryngium belongs to the Apiaceae family and includesaround 250 species that are widespread throughout the world.[1]

Among them, several Eryngium species have been used as orna-mental plants, condiments[2] or in traditional medicine.[3,4]

Eryngium maritimum L., usually named sea holly in England orPanicaut des mers in France, is a perennial plant (3060 cm high)with mauve owers (blossoming time, JuneSeptember), growingwild on the sandy beaches of western Europe, the Mediterraneanbasin and the Black Sea.[1] The plant is one of the typical dune

germacrene D (43.1%) and 9-muurolene-15-aldehyde (22.4%)as main components of the essential oil from the aerial parts,and g-guaiene (40.2%), trimethylbenzaldehyde (24.5%) andgermacrene D (10.6%) as main components of the essentialoil from the roots.[23] The second study reports spathulenol, 1,5-epoxysalvial-4(14)-ene, a-amorphene and caryophellene oxideas main compounds of the essential oil from the aerial parts.[24]

Previous laboratory investigations of E. maritimum essential oilallowed the isolation of a known sesquiterpene (4bH-muurol-9-en-15-al) and three new oxygenated sesquiterpenes (4bH-cadin-9-en-15-al, 4bH-muurol-9-en-15-ol and 4bH-cadin-9-en-15-ol) whichChemical variability andEryngium maritimum L.Corsica and SardiniaFlorent Darriet,a Stphane Andreani,Jean Costaa and Alain Musellia*

ABSTRACT: The chemical compositions of Corsican and Sardincolumn chromatography, gas chromatography with ame ionin electron impact mode. Sixty-three compounds were idegermacrene D (13.745.9%), three uncommon oxygenated seen-15-ol (2.214.3%) and 4bH-muurol-9-en-15-al (4.39.3%), wfrom the plant aerial parts. Relative to these, essential o2,4,5-trimethylbenzaldehyde (39.8%), 2,3,6-trimethylbenzaldeof Corsican and Sardinian E. maritimum essential oils was stuisland of origin of the sample essential oils and their chemical coamounts of hydrocarbon terpenes than Sardinian samplesE.maritimum essential oils exhibited original compositions with

Received: 19 June 2012, Revised: 4 February 2013,

(wileyonlinelibrary.com) DOI 10.1002/ffj.3160studies were incomplete in terms of sample origins, full chemicalcomposition and relative percentages of the components; onlythe main oil components were reported. The rst study reports

Flavour Fragr. J. 2013 Copyright 2013 JohnMarie-Ccile De Cian,b

Eryngiummaritimum L. essential oils were investigated usingtion detection and gas chromatographymass spectrometryed accounting for 85.895.7% of the total amount. Withuiterpenes, 4bH-cadin-9-en-15-al (18.427.6%), 4bH-cadin-9-e identied asmain components of the essential oils obtainedfrom the roots differed drastically with high contents ofe (29.0%) and a-muurolene (23.5%). The chemical variabilityd using statistical analysis. A direct correlation between thepositions was assessed. Corsican essential oils exhibited higherelative to other Eryngium species, Corsican and Sardiniansquiterpene aldehydes. These compounds, together with totalNaturels, BP 52, 20250, Corte, France

b Universit de Corse, UMR-CNRS 6134 SPE, Laboratoire de GntiqueMolculaire, BP 52, 20250, Corte, France

Wiley & Sons, Ltd.

antioxidants of lipid peroxidation, while the oxygenated terpeneshad a high hydrogen-donating capacity towards radicals.[11]

The aim of the present work was to obtain a better insightinto the nature of the E. maritimum volatiles by: (1) using columnchromatography (CC), gas chromatography (GC) and gas chroma-tographymass spectrometry (GC-MS) analysis of the essential oilsobtained from aerial parts and roots; (2) investigating intra-speciesvariations in the essential oils from six Corsican and six Sardiniansample locations and comparing the oils with those fromliterature data; and (3) determining the antioxidant activities byusing the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,20-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS)methodswith the entireoil and the separated uncommon oxygenated sesquiterpenes.

Materials and Methods

Chemicals

For measurement of response factors (RFs), the chemicals usedwere: neo-allo-ocimene, a-pinene, b-pinene, g-terpinene, limonene,

University of Corsica, Corte, France. The fresh plant material washydro-distilled (5 h) using a Clevenger-type apparatus accordingto the method recommended in the European Pharmacopoeia,[26]

and the yield of essential oil was 0.060.13% based on freshweight.

Oil Fractionation

Sample C1 (2 g) was submitted to column chromatography (CC)on a silica gel (column diameter 2 cm, 30 g of 200500 mm silica,height 11 cm, ow 6 mL/min). A hydrocarbon fraction (HF; 599mg) was obtained by elution with n-pentane and an oxygenatedfraction (OF; 1250 mg) was obtained by using pure diethyl oxide.The hydrocarbon fraction (500 mg) was further chromatographedon silica gel impregnated with silver nitrate (column diameter 1 cm,20 g of 63200 mm silica with 5 g of silver powder, height 22.5 cm,ow 2.5 mL/min), leading to 12 fractions by elution with n-pentane [HF1 (40 mg), HF2 (30 mg), HF3 (50 mg), HF4 (70 mg), HF5(70 mg), HF6 (60 mg), HF7 (40 mg), HF8 (30 mg), HF9 (30 mg), HF10

and carefully added to a suspension of aluminium lithium hydride(100mg) in dry diethyl ether (60mL) at 0C. Themixture was stirred

al

41 100 5900 N, 9 270 5200 E40 490 4800 N, 8 330 2400 E

F. Darriet et al.Plant Material and Isolation of the Essential Oil

Aerial parts of Eryngium maritimum L. were collected in full bloom(June 2009) from six stations in Corsica (C1C6) and six stations inSardinia (S1S6). The sample numbers, the localities of harvest andthe GPS coordinates are given in Table 1. For the study of thechemical composition of the essential oil from E. maritimumseparated organs, a supplementary harvest of stems, owers,leaves and roots was obtained from Quercioni location (C1). Avoucher specimen was deposited in the herbarium of the

Table 1. Data relative to the different samples of plant materi

No.a Localityb

C1 QuercioniC2 PiniaC3 MaranaC4 OstriconiC5 Golfe de VentilegneC6 CalzarelloS1 La CalettaS2 Isula di CapreraS3 Marina di SorsoS4 Golfo di S EnaS5 PraxisS6 CagliariaNumbers associated with the samples.bLocalities of the harvests.b-caryophyllene, a-humulene, aromadendrene, nerol, lavandulol,(E)-hex-3-en-1-ol, cedrol, globulol, pentyl acetate, lavandulylacetate, trans-myrtenyl acetate, cedryl acetate, artemisia ketone,camphor, jasmone, isoborneol methyl ether, carvacrol methylether, caryophyllene oxide, (E)-2-hexenal, (E,E)-2,4-decadienaland (E)-2-decenal. Authentic chemical samples were obtained fromSigmaAldrich (Saint-Quentin Fallavier, France) and Fluka (Saint-Quentin Fallavier, France) in the highest available purity. For theantioxidant screening, methanol, DPPH and ascorbic acid werepurchased from SigmaAldrich.Copyright 2013 Johnwileyonlinelibrary.com/journal/ffj39 480 5400 N, 8 330 000 E39 70 000 N, 9 310 1200 E39 120 3100 N, 9 50 1400 Eat room temperature and then reuxed for 3 h. The reaction mix-ture was hydrolysed by the addition of 15% sodium hydroxide so-lution (1 mL) and cold water. The organic layer was separated,washedwithwater to neutrality, dried over sodium sulfate and con-centrated under vacuum. After purication on CC the alcohol-richfraction (ROF1; 250 mg) contained 4bH-cadin-9-en-15-ol (63.8%)and 4bH-muurol-9-en-15-ol (33.5%) as major components.

GPS coordinates

41 560 4600 N, 9 240 4100 E42 10 2100 N, 9 280 3000 E42 390 1400 N, 9 270 1000 E42 390 3600 N, 9 30 3400 E41 280 3200 N, 9 40 4400 E41 590 100 N, 9 260 900 E40 350 3600 N, 9 450 2100 E(20 mg), HF11 (20 mg) and HF12 (20 mg)].The oxygenated fraction (1202 mg) was further

chromatographed on silica gel (column diameter 1 cm, 60 g of63200 mm silica, height 46.5 cm, ow 2 mL/min) with agradient pentanediethyl oxide (95/5) leading to six fractions[OF1 (300 mg), OF2 (180 mg), OF3 (130 mg), OF4 (130 mg),OF5 (200 mg) and OF6 (240 mg)].

Reduction of Oxygenated Fraction 1

Fraction OF1 (300 mg) was dissolved in dry diethyl ether (40 mL)Flavour Fragr. J. 2013Wiley & Sons, Ltd.

Mode

the polar column. For each sample, two reconstructed ionic

(Jenway, Bibby Scientic Limited, Staffordshire, UK). Inhibition of

azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical (ABTS+)

Eryngium maritimum, essential oils, antioxidant activityComponent quantication of the E. maritimum essential oils wascarried out using peak normalization, including RFs, with an inter-nal standard, and expressed as normalized % abundances. Thecomplexity of the oil, together with the lack of standards, makesdetermination of the RFs of all the components unrealistic. So, weused a solution introduced by Costa et al.[31] based on groupingthe essential oil components by their functional groups and thenby their chemical class. To calculate the RF of a compound forwhich a standard is not available, with another one, it is essentialthat the two compounds have the same empirical formula. RFsand calibration curves were determined by diluting each stan-dard (see chemicals) in dichloromethane, at ve concentrations,with each specimen containing tridecane (nal concentration0.7 g/100 g) as internal standard (IS); analyses were performedin triplicate. The response factors were calculated by using theequation RF = {C /[(A /A )]}C , where C ischromatograms were provided and they have been investigatedconsecutively. Other GC conditions were the same as describedabove. Ion source temperature, 150C; energy ionization, 70 eV;electron ionization mass spectra were acquired with a mass rangeof 35350 amu during a scan time 1 s. Oil injected volume, 0.1 mL;fraction injected volume, 0.2 mL.

Component Identication

Identication of individual components was based on: (1) acomparison of calculated retention indices, on polar and apolarcolumns, with those of authentic compounds or literaturedata;[27,28] and (2) on computer matching with commercial massspectral libraries and comparison of mass spectra with those ofour own library of authentic compounds or literature data.[2730]

Component QuanticationThe essential oils and the fractions obtained by CC wereinvestigated using a PerkinElmer TurboMass quadrupoledetector, directly coupled to a PerkinElmer AutoSystem XL(Walton, MA, USA) equipped with two fused-silica capillarycolumns (60 m 0.22 mm, lm thickness 0.25 mm), Rtx-1(polydimethylsiloxane) and Rtx-Wax (polyethylene glycol). Bothcolumns were used with the same MS detector. Oil analyseswere consecutively carried out on the apolar and then onGas Chromatography with Flame Ionization Detection

Gas chromatography with ame ionization detection (GC-FID)analyses were carried out using a PerkinElmer Clarus 600 GCapparatus (Walton, MA, USA) equipped with a single injectorand two ame ionization detectors. The apparatus was usedfor simultaneous sampling to two fused-silica capillarycolumns (60 m 0.22 mm, lm thickness 0.25 mm) withdifferent stationary phases: Rtx-1 (polydimethylsiloxane) andRtx-Wax (polyethylene glycol). The temperature programwas: 60230C at 2C/min and then held isothermal at230C for 30 min. The carrier gas was helium (1 mL/min).The injector and detector temperatures were held at 280C.Split injection was conducted with a ratio split of 1:80.Injected volume was 0.1 mL.

Gas ChromatographyMass Spectrometry in Electron Impactanalyte abs,analyte abs,IS IS analyte

Flavour Fragr. J. 2013 Copyright 2013 Johnwas evaluated according to the method of Re et al.[35] with mod-ications. ABTS+ was produced by reaction of equal volumes of2.4 mM ABTS solution and 2.4 mM potassium persulfate for 16 h,in the dark and at room temperature. The stock solution wasthen diluted in 0.2 M phosphate-buffered solution to achievean absorbance of 0.80 0.05 at 734 nm. Then 900 ml of dilutedABTS+ solution was mixed with 0.110 mL of sample dissolvedin 100 mL of methanol. The absorbance at 734 nm was taken 1min after mixing. Inhibition of free-radical oxidation wasexpressed in % and calculated according to the same equationas used for DPPH scavenging assay.the free radical, DPPH (I%) was calculated using the equation: I%=100 (AbAs)/Ab, where Ab is the absorbance of the controlreaction and As the absorbance of the sample. The sample concen-tration providing 50% inhibition (IC50) was calculated from thegraph of inhibition percentage against sample concentration.Tests were carried out in triplicate. Ascorbic acidwas used as a pos-itive control.

The 2,20-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical-scavenging assay

The ability of total essential oil and fractions to bleach the 2,20-the concentration of the standard compound, Aabs,analyte is itsabsolute peak area, Aabs,IS is the tridecane absolute peak areaand CIS is its concentration (0.7 g/100 g). The average RFsobtained for each standard compound within a chemical classare used as a correction factor specic for each chemical class.This procedure gave RFs relative to tridecane (1.01 for monoterpenehydrocarbons, 1.0 for sesquiterpene hydrocarbons, 1.34 for alcohols,1.55 for esters, 1.31 for ketones, 1.24 for ethers, 1.59 for oxides and1.40 for aldehydes) and allows the determination of the normalized% abundances using the methodology reported by Bicchi et al.[32]

Statistical Analysis

In order to study the chemical variations in E.maritimum essentialoils, a standardized data matrix was established from thechemical composition of the 12 Corsican and Sardinian samplesstudied in this present work. The data matrix displayed wasanalysed using Principal component analysis (PCA) and hierarchi-cal ascending classication[33] with the aid of XLSTAT software(Version 2012.2.04; Addinsoft, Paris, France). PCA was made witha Pearson matrix and hierarchical ascending classication wasmade with a Euclidian matrix and Ward aggregation.

Antioxidant Screening

The 2,2-diphenyl-1-picrylhydrazyl radical-scavenging assay

The hydrogen atoms or electron-donating ability of the totalessential oil and corresponding fractions were measured fromthe bleaching of purple-coloured methanol solution of DPPHas described by Sharifar et al.[34] with some modications. A100 mL aliquot of methanol containing amounts of E. maritimumsamples ranging from 100 ng to 10 mg was added to 1 mL of a0.2 mM methanol solution of DPPH. Ascorbic acid (125 mg/mLnal concentrations) was used as the antioxidant standard. Themixture was homogenized and incubated for 25 min at room tem-perature in the dark. The absorbance was measured at 515 nmagainst a blank using a 6405 UVvisible spectrophotometerWiley & Sons, Ltd. wileyonlinelibrary.com/journal/ffj

The total essential oil was dominated by 17 oxygenated sesqui-

F. Darriet et al.terpenes (49.8%) and 26 sesquiterpenes hydrocarbon (42.2%),whereas the 15 other compounds represent only 2.5% of the oil.The main components were germacrene D, 34 (33.9%), 4bH-cadin-9-en-15-al, 56 (26.1%), 4bH-cadin-9-en-15-ol, 58 (9.5%) and4bH-muurol-9-en-15-al, 55 (5.2%). The essential oil was character-ized by a large variety of sesquiterpene skeletons: acyclic(nerolidane), cyclic (bisabolane, bergamotane, germacrane andhumulane), bi-cyclic (bicyclogermacrane, caryophyllane, cadinaneand muurolane) and tri-cyclic (cubebane, copaane, bourbonane,ylangane and aromadendrane).

In addition, leaves, stems, owers and roots of E. maritimumharvested from the same location (C1; Quercioni, Corsica) affordedessential oils with similar yields (0.060.09%). Integrated analysis ofthe four sample essential oils allowed identication of 58compounds, which always accounted formore than 90.5%of the es-sential oils (Table 2). Essential oils from the leaves, stems, owers andfull aerial parts were qualitatively similar and exhibited only few dif-ferences in the percentages of their main components. Relative tothe essential oil from the aerial parts, the essential oil fromE. maritimum roots displayed the simplest gas chromatogram.Only eight compounds, which accounted for 97.4% of the oil,were identied, and among them 2,4,5-trimethylbenzaldehyde, 14(39.8%), 2,3,6-trimethylbenzaldehyde, 15 (29.0%) and a-muurolene,38 (23.5%) were the threemain components. Corsican root essentialoil was quite different to the root essential oil reported by Kubeczkaet al.[23] which contained trimethylbenzaldehyde (isomer notreported, 24.5%) and the sesquiterpene hydrocarbons g-guaiene(40.2%) and germacrene D (10.6%) as the main components.

Chemical Variability of E. maritimum Essential Oils and theComparison with Literature Data

Aerial parts of 12 E. maritimum oil samples, six from Corsica(C1C6) and six from Sardinia (S1S6), were hydrodistilledto afford essential oils with moderate yields: 0.060.13% offresh material. The essential oils were analysed to obtain abetter understanding of their chemical variability. GC-FIDand GC/MS-EI analyses of the 12 essential oils showed somequantitative differences between sample essential oils fromboth islands and the occurrence of ve additionalcompounds, denoted 10, 20, 70, 120 and 580, in the Sardiniansample of essential oils (Table 3).

The standardized essential oil matrix was statistically analysedemploying hierarchical ascending classication and principalcomponent analysis. The dendrogram and plot establishedResults and Discussion

Chemical Composition of Corsican E.maritimum Essential Oil

Due to the wide distribution of Eryngium maritimum along thesandy beaches of east Corsica, the station of Quercioni (C1)was chosen for detailed analysis. Preliminary analysis of the sam-ple of essential oil from the aerial parts of E. maritimum allowedthe identication of only 38 compounds, which accounted for94.5% of the C1 sample oil. Investigation of all CC fractions andsub-fractions by GC-FID and GC/MS-EI led to the identicationof 58 components which accounted for 94.5% of the sampleoil (Table 2). The chemical composition of both hydrocarbonand oxygenated CC fractions (HF and OF, respectively) arereported in Table 2. Oil components included ve monoter-penes, 43 sesquiterpenes and 10 non-terpenic compounds.Copyright 2013 Johnwileyonlinelibrary.com/journal/ffjusing the rst two axes, which accounted for 44.43% and20.79% of the total variance, suggest the existence of twoclusters (Figure 1 and Figure 2). Figure 2 shows the distributionof the discriminating volatile compounds germacrene D, 34,4bH-cadin-9-en-15-ol, 58, aromadendrene oxide, 50, and thedistribution of oil samples. The F1 axis is negatively correlatedwith oxygenated sesquiterpenes and positively correlated withsesquiterpene hydrocarbons.

Cluster I and cluster II include the Corsican and Sardinian sampleessential oils, respectively. The essential oils from both islands dif-fered by the relative amounts of their main components, especiallygermacrene D, 34, 4bH-cadin-9-en-15-ol, 58, and aromadendreneoxide, 50, as seen on PCA. The main components of SardinianE. maritimum samples were 4bH-cadin-9-en-15-al, 56 (19.726.1%),4bH-muurol-9-en-15-al, 55 (6.39.2%), 4bH-cadin-9-en-15-ol, 58(9.014.3%), germacrene D, 34 (13.723.8%) and aromadendreneoxide, 50 (1.25.0%). The main components of Corsican E.maritimum samples were germacrene D, 34 (33.145.9%), 4bH-cadin-9-en-15-al, 56 (18.526.1%), 4bH-cadin-9-en-15-ol, 58(5.29.5%) and 4bH-muurol-9-en-15-al,- 55 (5.2-8.3%). Sampleoils of both clusters were discriminated by the proportions of (1)terpene hydrocarbons, which were lower in the Sardinian sampleessential oils (21.638.2%) than in the Corsican sample essentialoils (39.056.9%); and (2) oxygenated terpenes, which were higherin Sardinian sample essential oils (62.647.5%) than in Corsicansample essential oils (38.649.9%). It is noteworthy that CorsicanE. maritimum oil was quite similar to that reported by Kubeczkaet al.[23] except for 4bH-cadin-9-en-15-al, 56, and 4bH-cadin-9-en-15-ol, 58, which were natural compounds identied for the rsttime in E. maritimum in our previous work.[25] In addition, theCorsican oil sample was radically different to those reported byAslan and Kartal[24] in which spathulenol, 1,5-epoxysalvial-4(14)-ene, a-amorphene and caryophellene oxide were identied asthe main components.

Relative to essential oils from other Eryngium species previouslystudied,[1223,3656] samples of E. maritimum studied here showsimilarity with E. campestre (Egyptian sample)[40] and E. duriaei[43]

in which sesquiterpene aldehydes are present in large amounts.They differed from the others. Eleven essential oil sampleswith high amounts of non-terpenic compounds, especiallytrimethylbenzaldehyde, linear aldehydes [such as (E)-2-dodecenal]and linear acids are reported in the literature. These Eryngiumsample essential oils were distributed in four species: E. foetidum(seven samples),[17,4550] E. corniculatum (two samples),[21]

E. creticum[39] and E. caucasicum (one sample).[41] Thirty-three essen-tial oil samples from 18 various species are dominated by terpenichydrocarbon compounds such as phyllocladene, a-pinene andgermacrene D. It is the most represented essential oil pattern inthe Eryngium genus in terms of species. The 17 species weredistributed as: E. billardieri,[37] E. glaciale (two samples),[15]

E. pandanifolium (two samples),[44] E. bourgatii (two samples),[14]

E. serbicum,[51] E. campestre (Turkish and eight Spanish sam-ples),[39,56] E. yuccifolium (two samples),[12] E. caeruleum,[42]

E. thorifolium,[39] E. paludosum,[54] E. vesiculosum,[44] E. aqualifolium(two samples),[36] E. expansum,[44] E. rosulatum,[55] E. amethystinum(three samples),[13] E. planum (steam+ leaf sample)[52] andE. caucasicum (one sample).[41] Finally, seven samples of essentialoils which comprise ve species distributed as E. bungei,[38]

E. palmatum,[51] E. rostratum (two samples),[44] E. paniculatum,[53]

E. foetidum (Cuban sample)[20] and E. planum (one sample)[52] aredominated by terpene alcohols or oxides, such as spathulenol,carotol or aromadendrene oxide.Flavour Fragr. J. 2013Wiley & Sons, Ltd.

Table

2.Che

mical

compo

sitio

nsof

CorsicanEryn

gium

maritimum

essentialo

ils(C1locatio

n,Que

rcioni),co

lumnch

romatog

raph

y(CC)fractio

nsan

dsepa

ratedorga

ns

No.

aCom

poun

dLR

IbRI

Ac

RIPd

RFse

Totalo

il,f

aeria

lparts

CCfractio

nsf,g

Sepa

ratedorga

nsf

Iden

ticatio

nh

HF

OF

Flow

ers

Stem

sLeaves

Roots

1a-Pine

ne93

693

099

41.01

0.1

0.6

0.1

0.6

tr

RI,M

S2

6-Methy

lhep

t-5-en

e-2-on

e97

296

715

701.31

tr

0.2

trtr

tr

RI,M

S3

2-Pe

ntyl-furan

981

975

1201

1.59

tr

trtr

trtr

RI,M

S4

Octan

al98

197

712

901.40

tr

trtr

trtr

RI,M

S5

Myrcene

987

983

1130

1.01

0.2

0.4

0.2

tr0.1

RI,M

S6

1,2,3-Trim

ethy

lben

zene

1011

1006

1294

1.01

tr0.1

tr

0.1

tr

RI,M

S,Re

f7

Limon

ene

1025

1019

1166

1.01

0.1

0.1

0.1

trtr

RI,M

S8

cis-Ve

rben

ol11

3211

2616

181.34

tr

trtr

tr

RI,M

S9

tran

s-Ve

rben

ol11

3211

3016

371.34

0.1

0.1

tr

0.1

RI,M

S10

(E)-Non

-2-ena

l11

3911

4413

941.40

0.1

0.1

tr0.1

tr

RI,M

S11

4-Methy

lacetoph

enon

e11

5611

5317

311.31

0.1

0.1

trtr

0.1

RI,M

S,Re

f12

Decan

al11

8011

8214

981.40

tr

trtr

0.1

tr

RI,M

S13

2,4,6-Trim

ethy

lben

zaldeh

yde

1280

1281

1827

1.40

tr

tr

tr0.1

RI,M

S14

2,4,5-Trim

ethy

lben

zaldeh

yde

1305

1297

1846

1.40

0.3

0.5

0.6

0.1

39

.8RI,M

S15

2,3,6-Trim

ethy

lben

zaldeh

yde

1314

1318

1935

1.40

1.5

1.9

2.8

0.9

tr29

.0RI,M

S16

a-Cub

eben

e13

5513

4614

521.0

tr0.2

0.1

trtr

trRI,M

S17

(Z)-b-Dam

asceno

ne13

4313

4918

201.31

0.1

0.2

0.1

0.1

tr

RI,M

S18

a-Cop

aene

1371

1373

1447

1.0

0.6

2.5

0.1

0.7

0.5

RI,M

S19

a-Ylan

gene

1372

1374

1476

1.0

tr0.1

0.1

tr

RI,M

S20

b-Bo

urbo

nene

1374

1375

1474

1.0

0.4

1.5

0.6

trtr

RI,M

S21

b-Elem

ene

1384

1385

1555

1.0

0.9

0.9

0.9

1.0

0.7

0.3

RI,M

S22

b-Pa

tcho

ulen

e13

8813

9414

751.0

tr0.1

tr

tr

RI,M

S23

b-Gurjune

ne14

0414

0515

911.0

trtr

tr

RI,M

S24

a-Gurjune

ne14

1314

1115

241.0

0.2

0.2

1.2

0.5

tr

RI,M

S25

cis-a-Be

rgam

oten

e14

1414

1614

801.0

1.0

4.3

1.0

1.8

0.7

RI,M

S26

b-Ylan

gene

1420

1420

1562

1.0

0.6

2.1

0.2

0.3

0.7

RI,M

S27

a-Se

squiph

elland

rene

1428

1430

1765

1.0

0.1

0.4

0.8

0.1

0.2

RI,M

S28

b-Cop

aene

1430

1431

1581

1.0

tr2.5

0.1

trtr

RI,M

S29

tran

s-a-Be

rgam

oten

e14

3414

3515

801.0

0.1

tr

0.1

0.1

0.1

RI,M

S30

Aromad

endren

e14

4314

4016

111.0

0.1

1.1

0.1

0.1

RI,M

S31

a-Hum

ulen

e14

5514

4916

651.0

0.1

1.3

0.6

trtr

RI,M

S32

g-Muu

rolene

1473

1468

1681

1.0

0.4

2.6

0.2

tr0.5

RI,M

S33

a-Curcu

men

e14

7414

7116

821.0

trtr

0.2

tr

RI,M

S34

GermacreneD

1479

1478

1659

1.0

33.9

47.7

32

.142

.532

.22.0

RI,MS

35b-Se

linen

e14

8614

8317

121.0

tr0.4

0.2

RI,M

S36

4-Ep

icub

ebol

1490

1487

1870

1.34

tr

0.6

tr0.1

RI,M

S37

Bicyclog

ermacrene

1494

1491

1979

1.0

0.3

1.9

3.0

2.2

tr

RI,M

S38

a-Muu

rolene

1496

1494

1719

1.0

1.1

2.6

tr1.2

23.5

RI,M

S39

b-Bisabo

lene

1503

1498

1720

1.0

1.1

1.7

0.2

0.6

1.5

RI,M

S

Eryngium maritimum, essential oils, antioxidant activity

Flavour Fragr. J. 2013 Copyright 2013 John Wiley & Sons, Ltd. wileyonlinelibrary.com/journal/ff40g-Cad

inen

e15

0715

0517

201.0

0.1

3.2

0.4

0.1

0.1

RI,M

Sj

Table

2.(Con

tinue

d)

No.

aCom

poun

dLR

IbRI

Ac

RIPd

RFse

Totalo

il,f

aeria

lparts

CCfractio

nsf,g

Sepa

ratedorga

nsf

Iden

ticatio

nh

HF

OF

Flow

ers

Stem

sLeaves

Roots

41Cub

ebol

1514

1509

1924

1.34

0.3

0.4

tr

0.4

RI,M

S42

d-Cad

inen

e15

2015

1317

001.0

1.2

13.4

2.5

0.9

0.6

RI,M

S43

Cad

ina-1,4-dien

e15

2315

2617

631.0

tr0.5

0.1

tr

RI,M

S44

(E)-Nerolidol

1553

1549

2037

1.34

0.5

1.1

2.1

0.4

0.2

RI,M

S45

Spathu

leno

l15

6915

6121

191.34

0.5

1

0.7

1.1

0.2

RI,M

S46

4a-Hyd

roxyge

rmacra-1,5-diene

1571

1565

2296

1.34

1.1

2.6

0.1

0.2

1.2

RI,M

S,Re

f47

Caryo

phyllene

oxide

1578

1570

1950

1.59

0.5

1.2

0.1

0.2

0.7

RI,M

S48

4b-Hyd

roxyge

rmacra-1,5-diene

1580

1571

2042

1.34

tr

0.5

0.2

trtr

RI,M

S,Re

f49

Muu

rola-4,10-dien

-8a-ol

1594

1597

2165

1.34

0.3

0.8

0.7

0.2

0.3

RI,M

S,Re

f50

Aromad

endren

eox

ide

1623

1618

2002

1.59

0.4

1.1

1.3

0.5

0.1

RI,M

S51

t-Muu

rolol

1633

1634

2143

1.34

0.8

2.4

0.6

0.4

1.0

RI,M

S52

t-Cad

inol

1633

1638

2163

1.34

0.4

0.9

1.8

0.5

0.2

RI,M

S53

a-Cad

inol

1633

1639

2227

1.34

1.5

3.2

1.5

2.3

1.3

RI,M

S54

Eude

sma-4,7-dien

e-1b

-ol

1671

1669

2354

1.34

2.2

5.6

trtr

3.0

1.6

RI,M

S,Re

f55

4bH-M

uurol-9-en

-15-al

16

8421

631.40

5.2

16

.35.2

9.3

4.3

RI,MS

564 b

H-Cad

in-9-en-15-al

16

8421

731.40

26.1

34

.718

.420

.327

.6

RI,MS

574b

H-M

uurol-9

-en-15

-ol

17

3424

221.34

0.4

0.6

0.2

0.3

0.4

RI,M

S58

4bH-Cad

in-9-en-15-ol

17

4224

521.34

9.5

20

.94.5

2.2

10.5

1.2

RI,MS

Totalide

ntied

f94

.594

.694

.190

.590

.990

.997

.4Hyd

rocarbon

compo

unds

42.6

92.3

49

.051

.639

.225

.8Oxyge

natedco

mpo

unds

51.9

2.3

94.1

41.5

39.3

51.7

71.6

Hyd

rocarbon

mon

oterpe

nes

0.4

1.1

0.4

0.6

0.1

Oxyge

natedmon

oterpe

nes

0.1

0.1

tr

0.1

Hyd

rocarbon

sesquiterpen

es42

.291

.2

48.6

51.0

39.1

25.8

Oxyge

natedsesquiterpen

es49

.82.3

91.2

38.1

38.0

51.5

2.8

Other

oxyg

enated

compo

unds

2.0

2.8

3.4

1.2

0.1

68.8

Other

Hyd

rocarbon

compo

unds

tr0.1

tr

0.1

Yields

%(v/dw)

0.08

0.09

0.08

0.06

0.09

a Order

ofelutionisgive

non

theap

olar

column(Rtx-1).Th

emainco

mpo

nentsareshow

nin

bold

type

.bRe

tentionindicesfrom

literatureon

theap

olar

column:

RIA,rep

ortedfrom

literature.[27,28]

c Reten

tionindiceson

theRtx-1ap

olar

column.

dRe

tentionindiceson

theRtx-wax

polarco

lumn.

eRe

spon

sefactors(RFs).Fo

rthecalculationmod

e,seetext.

f Percentag

esaregive

non

theap

olar

columnexcept

forco

mpo

unds

with

thesameRI

A(percentag

esaregive

non

thepo

larco

lumn);tr,pe

rcen

tage

sbe

low

0.05

.gFractio

nsob

tained

byco

lumnch

romatog

raph

y:HF,

hydroc

arbo

nfractio

n,OF,ox

ygen

ated

fractio

n(see

expe

rimen

tal).

hRI,reten

tionindices;MS,

massspectrain

electron

icim

pact

mod

e;Re

f,co

mpo

unds

iden

tied

from

commercial

data

librarie

s.[27]

F. Darriet et al.Flavour Fragr. J. 2013Copyright 2013 John Wiley & Sons, Ltd.wileyonlinelibrary.com/journal/ffj

Table

3.Che

mical

compo

sitio

nsof

Eryn

gium

maritimum

essentialo

ilsfrom

Corsica

andSardinia

No.

aCom

poun

dLR

IbRI

Ac

RIPd

RFse

Corsicansamples

Sardiniansamples

Iden

ticatio

ng

C1

C2

C3

C4

C5

C6

S1S2

S3S4

S5S6

10Santolinatrie

ne90

990

310

181.01

tr

0.2

RI,M

S1

a-Pine

ne93

693

099

41.01

0.1

0.5

0.4

tr0.4

0.1

0.1

0.2

0.4

0.4

0.2

1.3

RI,M

S2

6-Methy

lhep

t-5-en

e-2-on

e97

296

715

701.31

tr

0.1

tr0.1

0.1

0.1

0.2

0.1

0.2

tr0.2

RI,M

S20

Sabine

ne97

397

310

981.01

trtr

0.1

0.2

tr0.2

RI,M

S3

2-Pe

ntyl-furan

e98

197

512

011.59

trtr

0.2

trtr

trtr

trtr

trtr

0.2

RI,M

S4

Octan

al98

197

712

901.40

trtr

10.1

tr0.1

trtr

1.8

RI,M

S5

Myrcene

987

983

1130

1.01

0.2

0.6

trtr

0.7

tr0.1

0.2

tr0.1

0.2

trRI,M

S6

1,2,3-Trim

ethy

lben

zene

1011

010

0612

941.01

trtr

0.1

trtr

tr0.5

0.2

0.1

tr0.1

0.4

RI,M

S,Re

f7

Limon

ene

1025

1019

1166

1.01

0.1

0.1

0.2

tr0.1

tr1.0

0.2

0.1

0.9

0.4

0.2

RI,M

S70

Cam

phor

1123

1123

1517

1.31

0.6

RI,M

S8

cis-Ve

rben

ol11

3211

2616

181.34

trtr

trtr

trtr

trtr

0.1

tr0.1

0.2

RI,M

S9

tran

s-Ve

rben

ol11

3211

3016

371.34

0.1

tr0.1

trtr

tr0.2

tr0.2

0.3

0.1

1.2

RI,M

S10

(E)-Non

-2-ena

l11

3911

4413

941.40

0.1

tr0.1

0.2

tr0.1

0.2

0.1

0.2

tr0.1

0.5

RI,M

S11

Methy

l-4-acetoph

enon

e11

5611

5317

311.31

0.1

tr0.2

tr0.1

0.1

0.2

0.2

0.2

0.1

0.2

0.7

RI,M

S,Re

f12

Decan

al11

8011

8214

981.40

tr0.1

0.1

trtr

trtr

tr0.1

trtr

0.2

RI,M

S12

0tran

s-Chrysan

then

ylacetate

1238

1234

1494

1.55

0.1

1.7

2.1

1.5

0.4

0.4

RI,M

S13

2,4,6-Trim

ethy

lben

zaldeh

yde

1280

1281

1827

1.40

trtr

trtr

trtr

0.1

0.2

0.4

0.1

0.3

trRI,M

S14

2,4,5-Trim

ethy

lben

zaldeh

yde

1305

1297

1846

1.40

0.3

0.3

0.4

0.5

0.2

0.3

0.2

0.3

0.5

0.2

0.5

1.2

RI,M

S15

2,3,6-Trim

ethy

lben

zaldeh

yde

1314

1318

1935

1.40

1.5

2.0

1.4

2.6

1.0

1.1

2.8

1.8

2.0

2.5

1.9

6.2

RI,M

S16

a-Cub

eben

e13

5513

4614

521.0

trtr

trtr

trtr

trtr

trtr

trtr

RI,M

S17

(Z)-b-Dam

asceno

ne13

4313

4918

201.31

0.1

tr0.1

0.1

tr0.1

0.1

0.1

0.1

0.2

0.1

trRI,M

S18

a-Cop

aene

1371

1373

1447

1.0

0.6

0.5

0.8

0.6

0.9

0.8

0.7

0.4

0.8

0.4

0.9

0.7

RI,M

S19

a-Ylan

gene

1372

1374

1476

1.0

trtr

trtr

trtr

trtr

trtr

trtr

RI,M

S20

b-Bo

urbo

nene

1374

1375

1474

1.0

0.4

0.1

0.2

0.1

0.1

0.2

0.2

0.3

0.3

0.1

0.4

RI,M

S21

b-Elem

ene

1384

1385

1555

1.0

0.9

0.9

1.1

1.0

1.3

1.3

0.5

0.4

1.2

0.9

1.0

0.8

RI,M

S22

b-Pa

tcho

ulen

e13

8813

9414

751.0

trtr

trtr

tr0.6

trtr

RI,M

S23

b-Gurjune

ne14

0414

0515

911.0

tr

trtr

0.1

0.1

tr

0.1

RI,M

S24

a-Gurjune

ne14

1314

1115

241.0

0.2

tr0.6

0.7

0.7

0.6

0.9

1.1

0.8

10.9

RI,M

S25

cis-a-Be

rgam

oten

e14

1414

1614

801.0

1.0

tr1.7

0.7

1.8

1.0

2.4

d1.2

1.2

1.0

1.5

RI,M

S26

b-Ylan

gene

1420

1420

1562

1.0

0.6

tr1.0

0.2

0.7

0.9

0.1

0.2

0.2

0.9

RI,M

S27

a-Se

squiph

elland

rene

1428

1430

1765

1.0

0.1

tr0.1

0.8

tr1.0

0.2

0.1

0.2

0.1

0.1

0.3

RI,M

S28

b-Cop

aene

1430

1431

1581

1.0

tr

0.2

tr0.5

tr

trRI,M

S29

tran

s-a-Be

rgam

oten

e14

3414

3515

801.0

0.1

0.2

tr0.2

0.1

0.1

RI,M

S30

Aromad

endren

e14

4314

4016

111.0

0.1

trtr

0.2

0.2

0.2

0.1

0.2

0.4

0.4

0.2

RI,M

S31

a-Hum

ulen

e14

5514

4916

651.0

0.1

tr0.3

0.2

0.3

0.3

0.4

0.1

0.1

0.1

0.1

0.2

RI,M

S32

g-Muu

rolene

1474

1468

1681

1.0

0.4

0.1

0.2

0.1

trtr

0.1

0.3

0.3

0.1

trtr

RI,M

S33

a-Curcu

men

e14

7314

7116

821.0

trtr

trtr

trtr

0.1

0.5

0.2

0.4

RI,M

S34

Germacrene

D14

7914

7816

591.0

33.9

33.1

32.2

43.5

45.9

41.4

13.7

14.6

16.6

14.3

23.8

18.4

RI,M

S35

b-Se

linen

e14

8614

8317

121.0

trtr

0.2

trtr

0.3

0.2

0.2

0.1

0.2

0.1

trRI,M

S36

4-Ep

icub

ebol

1490

1487

1870

1.34

trtr

tr0.3

0.2

tr

0.3

trRI,M

S

Eryngium maritimum, essential oils, antioxidant activity

Flavour Fragr. J. 2013 Copyright 2013 John Wiley & Sons, Ltd. wileyonlinelibrary.com/journal/ffj

Table

3.(Con

tinue

d)

No.

aCom

poun

dLR

IbRI

Ac

RIPd

RFse

Corsicansamples

Sardiniansamples

Iden

ticatio

ng

C1

C2

C3

C4

C5

C6

S1S2

S3S4

S5S6

37Bicyclog

ermacrene

1494

1491

1979

1.0

0.3

1.5

1.8

2.3

2.3

2.3

0.2

0.6

1.2

0.9

1.2

0.8

RI,M

S38

a-Muu

rolene

1496

1494

1719

1.0

1.1

0.2

0.1

0.1

0.1

0.1

0.4

0.4

0.3

0.2

trRI,M

S39

b-Bisabo

lene

1503

1498

1720

1.0

1.1

0.1

0.1

tr0.1

0.1

0.2

0.1

0.2

0.1

0.2

RI,M

S40

g-Cad

inen

e15

0715

0517

201.0

0.1

0.4

0.4

0.4

0.3

0.4

0.3

0.3

0.3

0.4

0.2

0.3

RI,M

S41

Cub

ebol

1514

1509

1924

1.34

0.3

tr

tr

0.1

tr0.1

0.2

tr

RI,M

S42

d-Cad

inen

e15

2015

1317

001.0

1.2

1.2

1.4

1.6

1.2

1.5

0.5

0.6

1.0

0.8

0.7

1.1

RI,M

S43

Cad

ina-1,4-dien

e15

2315

2617

631.0

tr

0.1

tr

0.2

0.1

0.3

0.2

0.2

0.1

RI,M

S44

(E)-Nerolidol

1553

1549

2037

1.34

0.5

0.2

0.8

0.5

tr0.4

0.7

0.5

1.0

0.7

0.6

1.5

RI,M

S45

Spathu

leno

l15

6915

6121

191.34

0.5

1.5

0.3

0.1

0.1

1.1

0.9

1.3

1.7

1.5

1.2

0.1

RI,M

S46

4a-Hyd

roxyge

rmacra-1,5-diene

1571

1565

2296

1.34

1.1

0.3

2.2

0.6

0.9

0.6

1.9

1.1

0.7

0.5

0.4

3.6

RI,M

S,Re

f47

Caryo

phyllene

oxide

1578

1570

1950

1.59

0.5

0.2

1.4

0.1

0.4

0.2

0.8

0.6

0.5

0.3

0.3

2.1

RI,M

S48

4b-Hyd

roxyge

rmacra-1,5-diene

1580

1571

2042

1.34

tr0.2

trtr

tr

0.4

tr0.4

0.2

0.2

RI,M

S,Re

f49

Muu

rola-4,10-dien

-8a-ol

1594

1597

2165

1.34

0.3

0.7

0.7

0.4

0.1

0.4

0.4

0.8

0.7

0.4

0.2

0.1

RI,M

S,Re

f50

Aromad

endren

eox

ide

1623

1618

2002

1.59

0.4

tr0.6

0.2

tr0.4

2.9

5.0

1.2

1.9

2.0

1.2

RI,M

S51

t-Muu

rolol

1633

1634

2143

1.34

0.8

1.2

0.5

0.8

tr0.6

1.8

5.8

1.0

1.6

2.1

0.6

RI,M

S52

t-Cad

inol

1633

1638

2163

1.34

0.4

0.6

1.2

1.3

tr0.3

2.3

1.3

2.0

2.2

0.8

0.9

RI,M

S53

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s.[27]

F. Darriet et al.

Flavour Fragr. J. 2013Copyright 2013 John Wiley & Sons, Ltd.wileyonlinelibrary.com/journal/ffj

bicyclo[4.4.0]decene aldehydes have not been reported inother species of Eryngium genus essential oils and it would beinteresting to check the presence of these compounds in theE. maritimum essential oils from different origins in order toconsider them as chemical markers of the species.

Antioxidant Activity of E. maritimum Essential Oil andFractions

Antioxidant properties of E. maritimum essential oil and bothhydrocarbon and oxygenated fractions obtained by columnchromatography were tested regarding their scavenging activitieson DPPH and ABTS radicals. Total essential oil showed a strong

Eryngium maritimum, essential oils, antioxidant activityFigure 1. Dendrogram of essential oil chemical compositions of Corsicanand Sardinian Eryngium maritimum samplesAs a concluding remark in this section, it is noticeable thatquantitative chemical variability between Corsican and SardinianE. maritimum samples oil reported here is probably linked to theharvest area. Moreover, it would be interesting to examinethe genetic diversity of both Corsican and Sardinian populationsin order to dene the origin of the essential oil variability.Compared to other species, E. maritimum sample essential oilswere clearly discriminated by the higher amount of sesquiter-pene aldehydes, such as 4bH-cadin-9-en-15-al, 56 (18.526.1%)and 4bH-muurol-9-en-15-al, 55 (5.29.2%). These uncommon

Figure 2. Principal component analysis of essential oil chemical compo-sitions of Corsican and Sardinian Eryngium maritimum samples

Table 4. Antioxidant activities of total essential oil and fractions oABTS radical-scavenging activity

Compoundused in an-tioxidantscreening

IC50 v

EO OF OF1

DPPH 5.9 0.1 8.8 0.1 111 3 31ABTS 0.7 0.03 1.34 0.02 29.9 0.8Results are given as mean SD (n= 3).EO, essential oil; OF, oxygenated fraction; OF1, fraction with uncom5758; HF, hydrocarbon fraction; AA, ascorbic acid (standard).

Flavour Fragr. J. 2013 Copyright 2013 John.3 219 38 9 347.7 35.2 6.1 0.04465.8 137.8 137.3 123.2 2.66 0.01

mon aldehydes 5556; ROF1, fraction with uncommon alcoholsantioxidant activity, since IC50 values regarding DPPH and ABTSradical-scavenging abilities, respectively 5.9 0.1 mg/mL and0.7 0.03 mg/mL, were equivalent or even lower than thoseregistered for ascorbic acid, used as the antioxidant reference(6.1 0.04 mg/mL for DPPH and 2.66 0.01 mg/mL for ABTS radicals,Table 4). Total essential oil was thus separated into two fractions,containing oxygenated or hydrocarbon compounds. The bestradical-scavenging effect was observed for the oxygenatedfraction, with IC50 values of 8.8 0.01 mg/mL against DPPHand 1.34 0.02 mg/mL against the ABTS radical, similar to thosefound for ascorbic acid and total essential oil. No signicantantioxidant activity could be detected in the hydrocarbon fraction,since IC50 values of this fraction were more than 50-fold higherthan those found for ascorbic acid, even though IC50 valuesregarding the ABTS radical were greater than those observedfor the DPPH radical (Table 4 and Figure 3). Considering theseresults, we could infer that the antioxidant properties exhibitedby E. maritimum essential oil are carried out by compounds of theoxygenated fraction. According to Table 1, the oxygenated fractionwas dominated by two aldehydes and their corresponding alcohols(4bH-cadin-9-en-15-al, 56, 4bH-muurol-9-en-15-al, 55, 4bH-cadin-9-en-15-ol, 58 and 4bH-muurol-9-en-15-ol, 57) representing,respectively, 34.7%, 16.3%, 20.9% and 0.6% of the fraction. Inorder to selectively examine the antioxidant properties of thesefour uncommon compounds, DPPH and ABTS radical-scavengingactivity tests were also performed on: (1) the sesquiterpenealdehyde-rich fraction (OF1) with 55 (60.8%) and 56 (31.9%)obtained by CC, and (2) sesquiterpene alcohol-rich fraction(ROF1) with 57 (63.8%) and 58 (33.5%) obtained by reduction ofOF1 (see experimental) (Table 4 and Figure 3). The aldehyde-richfraction (OF1) showed moderate antioxidant activity on the ABTSradical (IC50 = 29.9 0.8 mg/mL) but its effect on the DPPH radicalwas less (IC50 = 111.3 31.3 mg/mL). The corresponding alcohol-

f Eryngium maritimum in DPPH radical-scavenging activity and

alue (mg/mL) SEMROF1 HF AAWiley & Sons, Ltd. wileyonlinelibrary.com/journal/ffj

OF1, fraction with uncommon aldehydes, 5556; ROF1, fraction with

F. Darriet et al.rich fraction (ROF1) exhibited no signicant antioxidant properties,considering that the IC50 values were more than 30-fold higherthan those found for ascorbic acid (Table 4 and Figure 3).

The antioxidant properties of E. maritimum essential oil wasattributed to some oxygenated compounds contained in theoxygenated fraction but they do not seem to be directly correlatedwith the main sesquiterpenes, 5558. On the one hand, theintermediate antioxidant activity found for the aldehydes maincompounds and the absence of activity found for their relatedprimary alcohols when analysed separately could mean thatantioxidant activity requires synergic associations between thesemain compounds and other (minor) compounds of the oxygen-ated fraction. On the other hand, this fraction also exhibited sev-eral tertiary alcohols, such as eudesma-4,7-diene-1b-ol, 54 (5.6%),a-cadinol, 53 (3.2%), 4a-hydroxygermacra-1,5-diene, 46 (2.6%)t-muurolol, 51 (2.4%) and spathulenol, 45 (1%) that could, evenif in lower proportions, participate in the antioxidant activityof E. maritimum essential oil. Indeed, antioxidant propertieshave already been reported for essential oils displaying similarsesquiterpene composition[57] and the difference of the skeletonbetween tertiary and primary alcohols could explain their reactivity

uncommon alcohols, 5758; HF, hydrocarbon fractionFigure 3. Antioxidant activities of total essential oil and fractions ofE. maritimum. Black bars, DPPH radical-scavenging activity; grey bars,ABTS radical-scavenging activity. Data are given as mean SD (n=3).AA, ascorbic acid (standard); EO, essential oil; OF, oxygenated fraction;toward the ABTS and DPPH radicals.

Acknowledgements

The authors are indebted to the Agence Economique deDveloppement de la Corse (ADEC-CTC) and EuropeanCommunity (PIC INTERREG IIIA) for partial nancial support.

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Eryngium maritimum, essential oils, antioxidant activityFlavour Fragr. J. 2013 Copyright 2013 John Wiley & Sons, Ltd. wileyonlinelibrary.com/journal/ffj