Chromatographia

, Volume 69, Issue 5–6, pp 575–585 | Cite as

Volatile Constituents of Achillea ligustica All. by HS-SPME/GC/GC-MS. Comparison with Essential Oils Obtained by Hydrodistillation from Corsica and Sardinia

  • Alain Muselli
  • Marta Pau
  • Jean-Marie Desjobert
  • Marcia Foddai
  • Marianna Usai
  • Jean Costa
Full Short Communication

Abstract

The volatile components extracted from the headspace (HS) of Achillea ligustica All. samples and their separated organs using solid phase microextraction (SPME) were investigated by gas chromatography and gas chromatography-mass spectrometry. Fiftyseven compounds were identified, the main components were camphor (14.2–29.8%), artemisia ketone (0.3–26.7%), santolina alcohol (0.5–9.4%), camphene (3.0–9.0%) and trans-sabinyl acetate (1.6–5.5%). Moreover, the chemical composition of Corsican and Sardinian A. ligustica oils obtained from flowers and leafy stems harvested in four regions of both islands, were investigated. Two collective oils of A. ligustica were also investigated, comparison between both oils as well as from data literature were reported. A comparison of hydrodistillation and HS-SPME extraction of volatile components in term of isolation time, plant-consuming and chemical composition was discussed. HS-SPME technique was clearly fast in contrast to hydrodistillation (90 min/300 min). HS extraction was performed with a much smaller amount of plant than hydrodistillation. Although the aromatic profiles of HS-fractions and oils showed several quantitative differences HS-SPME can be applied to routine control analysis of aromatic and medicinal plants.

Keywords

Gas chromatography-mass spectrometry Headspace-solid phase microextraction Essential oils Asteraceae Achillea ligustica 

1 Introduction

The genus Achillea (Asteraceae) includes more than 80 species widespread all around the northern hemisphere. Among them, A. ligustica All. is a perennial, pubescent herbaceous plant of the Mediterranean region. The plant is 20–90 cm, high with pinnati-partite leaves, a slightly aromatic scent, and a typically bitter taste. Flowers are arranged in flat-topped clusters, and both disk and ray flowers are small and white [1, 2]. Different species of this genus were widely used in traditional medicine, especially Yarrow (A. millefolium), the most significant species of the genus has been used against inflammations and spasms during gastrointestinal disorders or for their positive effects on wound healing and hemorrhages [3]. In the Sardinian traditional medicine, A. ligustica was used in decoctions as an anthelmintic, against gastric pains and neuralgias, and as an anti-inflammatory on skin diseases [4]. In Corsica, the plant called “Erba Santa” (Holy grass) was used in cataplasms to relieve sprains and insect bites, the leaves had also a reputation for stopping haemorrhages and the plant was added to the fritters of the Holy Friday [5].

Extracts of A. ligustica have been widely studied. Several works reported on the isolation and structure elucidation of piperidine amides, sesquiterpene lactones with rare 5/6/5 skeletons, guainolides and flavonoids [6, 7, 8, 9, 10]. The occurrence of some pharmacologically active compounds in the methanolic extracts of A. ligustica has been reported [11]. The chemical composition of the essential oils of species belonging to the genus Achillea have also been widely studied [12] and five studies reported the chemical composition of A. ligustica oil. An oil sample obtained from the aerial parts of Corsican A. ligustica [13], was characterized by camphor (21%) and santolina alcohol (19%) while eight samples of Sardinian A. ligustica oils exhibited different quantitative compositions, in which santolina alcohol (6%–21%), borneol (3%–20%), sabinol (2%–15%), trans-sabinyl acetate (1%–17%) and α-thujone (1%–25%) were identified as main components [14]. In another study of A. ligustica sample oil, terpineol-4 (19%), carvone (9%) and γ-terpinene (7%) were identified as major compounds in the leaves and linalool (20%) in the flowers [15]. Conversely, the major components of a continental Italian oil extracted from the aerial parts of A. ligustica, were artemisia acetate (44%), 2,7-dimethyl-4,6-octadien-2-ol (16%) and linalool (10%) [16]. Finally, linalool was reported as the main component of the leaves and flowers oils (28% and 71% respectively) from Grecian A. ligustica [17]. The Sardinian oil exhibited low antioxidant and antibacterial activities [14], while the oil from Corsica showed high antimicrobial activity against streptomyces [13].

The emitted volatile fraction plays a fundamental role in the plant’s life because it is an important biosensor diagnostic of the changes that take place in its metabolism. For the study of the plants volatile components, it is therefore necessary to develop analytical methods and techniques suitable both to detect variations in the composition of the emitted volatile fraction and to monitor the dynamics of the reactions of a vegetable organism when stressed [18]. Over the last ten years, there has been a remarkable renewal of interest in headspace sampling in particular after the introduction of high concentration capacity techniques (HCC) in which the recovery of volatiles is mainly based on the sorption approach [19]. Headspace solid phase microextraction (HS-SPME) is a rapid and simple procedure successfully used to sampling the volatile components from aromatic and medicinal plants [20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30]. Three studies reported the volatile components of Achillea millefolium (yarrow) extracted by HS-SPME [28, 29, 30]. The HS-sampling of volatile constituents need the optimization of parameters including fibers, times and temperatures of equilibrium and extraction. As reported in the literature [26], the most effective fiber used for the sampling of volatile compounds from vegetable matrices were generally those consisting of three polymers, a liquid (PDMS) for the less polar components and two solids (DVB and CAR) for the more polar components. Concerning times and temperatures of equilibrium and extraction, various conditions were reported according the plant material [20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30]. In the present work, the temperatures and extraction times used for the HS-sampling were optimized.

The aim of the present work was the characterization of the volatile fraction obtained from full aerial parts and different organs (flowers and leafy stems) of fresh A. ligustica using HS-SPME/GC-FID/GC-MS. The chemical compositions of Corsican and Sardinian essential oils obtained from flowers and leafy stems harvested in both islands were also investigated and a comparison between these and the literature data was reported. Moreover, a comparison of both volatile components extracted by SPME and HD from the same plant material was established. To the best of our knowledge, the components sampling from the vapour phase in equilibrium of A. ligustica using HS-SPME, was reported for the first time.

2 Experimental

2.1 Plant Material and Oil Isolation

The aerial parts of Achillea ligustica were collected at flowering stage, at five Corsican localities (France): Col St-Georges (A), Ghisoni (B), Cervioni (C), Propriano (D) and Corti (E) and four Sardinian localities (Italy): Monte Spada (F), Sant’Anna (G), Monte Limbara (H) and Baddeurbara (I). Voucher specimens were deposited in the Herbarium of the Universities of Corsica and Sassari. The aerial parts were separated in order to analyze separately the volatile components of flowers and leafy stems of each sample. The oils were isolated by hydrodistillation (110–190 g of plant per sample) for 5 h using a Clevenger-type apparatus [31] according to the European Pharmacopoeia and yielded 0.13–0.90% of a blue oil (see Table 1).
Table 1

Chemical composition (%) of Corsican and Sardinian Achillea ligustica oils

 

Components

lRIa

RIa

RIp

Corsican samples

Coll. oil

Flowers

Leafy stems

A

B

C

D

E

A

B

C

D

E

1

Nonene

886

880

930

0.1

0.1

0.1

0.1

0.1

0.1

0.1

0.1

0.1

0.1

0.4

2

Santolinatriene

909

901

1018

1.0

0.4

1.8

1.4

2.2

0.7

0.2

1.1

0.1

0.2

2.2

3

Tricyclene

927

921

1005

0.1

0.1

0.1

0.1

0.1

0.1

0.1

0.1

0.1

0.2

4

α-Tthujene

932

921

1008

0.1

0.1

0.1

0.1

0.1

0.1

0.1

5

α-Pinene

936

931

1008

0.9

1.2

1.3

1.0

1.0

0.9

0.7

1.0

0.1

0.2

2.4

6

Camphene

950

944

1045

1.6

2.6

2.2

2.0

1.3

1.4

1.3

1.7

0.5

3.3

7

Sabinene

973

965

1106

0.7

0.3

1.0

1.1

0.1

1.6

0.3

1.1

0.1

1.6

8

β-Pinene

978

971

1090

3.3

3.3

3.5

1.5

7.2

3.7

0.7

2.4

0.2

2.5

5.2

9

Myrcene

987

975

1143

0.1

0.1

0.2

0.1

0.1

0.2

0.1

0.1

10

Yomogi alcohol

991

984

1390

0.8

0.9

0.6

0.8

1.0

0.3

0.8

0.5

2.3

2.3

0.5

11

α-Phellandrene

1002

998

1142

0.1

0.1

tr

tr

tr

tr

0.1

0.1

0.1

0.1

12

α-Terpinene

1013

1010

1156

0.5

0.7

0.8

0.5

0.3

0.5

0.9

0.3

0.1

0.1

0.9

13

p-Cymene

1015

1013

1245

0.8

1.2

1.1

0.6

1.5

0.3

2.8

2.9

0.4

0.5

0.5

14

Limonene

1023

1021

1180

0.2

0.2

0.1

0.1

0.1

tr

0.2

0.1

0.1

0.1

tr

15

β-Phellandrene#

1023

1021

1186

16

1,8-Cineole

1023

1021

1196

1.5

1.5

1.8

3.0

1.0

3.4

1.3

2.1

3.9

0.3

0.4

17

Santolina alcohol

1029

1021

1385

15.4

3.3

11.5

10.6

30.1

3.8

2.9

10.6

9.4

22.7

10.1

18

cis-β-Ocimene#

1029

1030

1215

0.1

0.1

0.1

0.1

0.1

tr

0.1

tr

0.1

0.1

tr

19

trans-β-Ocimene#

1041

1040

1239

0.1

0.1

0.1

0.1

0.1

tr

0.1

0.1

0.1

tr

20

Artemisia ketone

1044

1046

1325

6.1

13.7

6.9

6.3

7.2

5.9

10.1

4.1

17.7

10.5

3.2

21

γ-Terpinene

1051

1050

1220

0.9

2.1

1.0

1.1

0.6

1.0

2.1

0.6

0.4

0.3

1.7

22

Isolyratol#

1052

1053

1524

23

trans-Sabinene hydrate

1053

1054

1440

0.3

0.4

0.5

0.4

0.1

0.1

0.6

0.9

0.9

0.1

4.2

24

Artemisia alcohol

1073

1069

1478

0.3

0.5

0.6

0.3

0.4

0.2

0.6

0.5

0.8

0.8

0.6

25

Terpinolene

1082

1079

1258

0.9

0.3

0.2

0.2

0.1

0.2

0.5

0.2

0.1

0.1

0.3

26

Linalool

1085

1084

1527

0.8

6.0

3.5

2.6

2.4

11.6

0.4

0.4

0.6

0.2

1.1

27

Filifolone

1086

1085

1419

28

α-Thujone

1089

1087

1402

0.5

0.7

29

β-Thujone

1103

1093

1422

1.0

0.9

1.1

0.1

1.0

0.4

0.6

0.1

0.4

30

Chrysanthenone

1110

1101

1480

31

cis-p-Menth-2-en-1ol

1108

1108

1533

32

Camphor

1123

1123

1490

20.8

26.2

13.5

15.4

8.5

17.2

22.0

12.7

3.9

14.7

17.0

33

trans-Sabinol

1120

1123

1664

4.2

2.5

5.1

7.0

0.1

7.4

0.5

7.1

2.3

0.1

1.5

34

Pinocarvone

1137

1140

1558

0.2

0.5

0.5

0.2

0.1

0.2

0.5

0.6

0.5

0.3

0.4

35

cis-Chrysanthenol

1147

1146

1716

36

Borneol

1150

1153

1664

7.8

8.5

2.9

2.4

2.1

2.6

3.8

2.1

0.7

3.0

1.3

37

Artemisyl acetate

1153

1155

1390

0.4

0.1

0.6

0.6

1.1

tr

0.1

0.6

1.7

1.0

tr

38

Terpinen-4-ol

1164

1164

1570

2.8

3.9

1.8

2.3

0.4

2.7

9.7

4.9

5.2

1.0

3.2

39

Myrtenal

1172

1173

1612

0.6

0.8

0.1

0.1

0.1

0.1

1.0

0.2

0.9

0.4

0.2

40

α-Terpineol

1176

1179

1664

0.2

0.2

0.1

0.4

0.2

0.7

0.6

0.4

0.6

0.5

0.6

41

Myrtenol

1178

1176

1772

42

cis-Piperitol#

1181

1181

1641

43

trans-Piperitol

1193

1191

1709

44

Piperitone

1226

1229

1696

45

cis-Chrysanthenyl acetate

1253

1244

1544

0.5

0.5

0.5

0.7

0.3

0.3

0.5

0.8

0.1

0.5

0.3

46

Bornyl acetate

1270

1271

1548

5.9

4.0

3.6

1.9

2.1

2.1

4.1

2.3

0.8

4.8

2.0

47

Thymol

1267

1272

2145

48

trans-Sabinyl acetate

1278

1274

1622

1.7

0.2

13.5

16.6

1.1

10.2

0.6

16.5

9.5

2.4

7.6

49

cis-Carvyl acetate

1318

1315

1710

0.2

0.1

0.2

0.2

0.1

0.2

0.2

0.1

0.3

0.3

0.2

50

Eugenol

1331

1329

2159

0.3

0.2

0.1

0.1

0.1

0.4

0.1

0.2

0.1

0.1

51

trans-Carvyl acetate

1345

1340

1749

tr

tr

0.1

0.1

tr

0.1

tr

0.1

tr

0.2

0.1

52

α-Cubebene#

1355

1346

1435

tr

tr

tr

0.1

tr

tr

tr

tr

tr

tr

tr

53

α-Longipinene#

1360

1350

1457

tr

tr

tr

tr

tr

tr

tr

tr

tr

tr

54

α-Ylangene#

1376

1369

1458

0.1

tr

tr

0.1

0.1

tr

tr

tr

0.1

tr

55

α-Copaene

1379

1374

1470

0.3

tr

tr

0.1

0.2

tr

tr

0.1

0.3

0.2

tr

56

β-Bourbonene#

1379

1386

1506

57

β-Elemene

1389

1385

1570

tr

tr

0.10

tr

0.1

tr

tr

0.2

0.2

0.1

tr

58

α-Gurjunene

1413

1408

1507

0.1

tr

tr

tr

tr

tr

tr

tr

tr

tr

tr

59

β-Caryophyllene

1421

1424

1578

0.2

0.3

0.10

0.2

0.1

tr

0.1

0.1

0.5

0.1

0.4

60

β-Copaene#

1430

1425

1565

0.1

tr

tr

tr

tr

tr

tr

tr

0.1

tr

tr

61

Aromadendrene

1443

1436

1597

tr

tr

tr

tr

tr

tr

tr

tr

tr

62

trans-β-Farnesene

1446

1448

1629

0.2

0.2

0.1

0.1

0.1

0.1

0.4

0.1

0.2

0.8

0.1

63

Alloaromadendrene

1462

1462

1620

0.4

0.3

0.4

0.1

0.7

0.4

0.6

0.3

0.9

0.5

0.6

64

α-Curcumene

1473

1473

1750

0.3

0.4

tr

0.3

0.3

tr

0.2

0.4

tr

65

γ-Curcumene#

1472

1474

1669

tr

tr

tr

tr

tr

tr

tr

tr

tr

tr

66

γ-Muurolene

1474

1474

1664

67

Germacrene-D

1479

1480

1682

0.9

2.0

1.4

tr

0.1

1.5

8.7

1.6

6.0

0.1

3.2

68

β-Selinene#

1486

1483

1695

0.2

0.1

tr

tr

tr

tr

tr

tr

tr

69

Zingiberene

1489

1489

1698

0.2

tr

tr

0.1

0.1

tr

tr

0.10

tr

70

Bicyclo germacrene

1494

1492

1710

0.1

0.2

0.1

0.2

0.3

0.2

0.6

0.1

0.3

0.2

0.3

71

Ledene

1491

1492

1669

0.2

tr

tr

tr

tr

tr

tr

tr

tr

tr

tr

72

α-Muurolene

1496

1494

1699

0.2

tr

tr

tr

tr

tr

tr

tr

tr

tr

73

trans, trans-α-Farnesene#

1498

1499

1730

0.2

tr

0.1

tr

tr

0.3

tr

0.8

tr

0.3

74

γ-Cadinene

1507

1507

1729

0.1

tr

tr

0.1

tr

0.1

0.1

tr

75

Calamenene

1517

1512

1802

0.10

tr

-

tr

tr

tr

tr

tr

tr

76

δ-Cadinene

1520

1515

1729

0.5

0.4

0.1

0.7

0.4

0.6

0.8

0.3

0.5

0.2

0.9

77

Cadina-1,4-diene#

1523

1523

1763

tr

tr

tr

tr

tr

tr

tr

tr

tr

tr

78

α-Calacorene

1527

1531

1880

0.1

0.1

0.1

tr

tr

0.1

0.2

tr

0.3

0.1

0.2

79

trans-Nerolidol

1533

1546

2031

0.1

tr

80

Elemol

1541

1536

2060

tr

tr

0.2

tr

tr

0.6

tr

0.1

0.2

0.4

81

β-Calacorene#

1566

1548

1913

tr

tr

tr

tr

tr

tr

tr

tr

tr

82

Spathulenol

1572

1560

2101

0.3

0.1

0.1

0.2

0.6

0.2

0.4

0.4

0.4

0.5

0.3

83

Palustrol

1569

1563

1903

tr

tr

0.10

0.1

0.1

tr

0.2

0.2

0.3

0.1

tr

84

Caryophyllene oxide

1578

1570

1960

tr

0.1

0.1

0.1

0.7

0.1

0.3

0.1

0.2

0.5

0.2

85

Viridiflorol

1592

1580

2039

7.9

4.4

5.5

5.3

14.4

8.2

5.4

5.1

13.4

15.0

9.5

86

Ledol

1600

1593

1983

0.6

0.6

0.4

0.4

0.7

0.6

0.4

0.4

0.9

1.7

0.6

87

epi-Cubenol

1623

1614

2032

0.3

0.3

0.5

0.5

0.1

0.6

0.5

0.5

1.0

0.6

0.5

88

γ-Eudesmol

1618

1622

2170

tr

0.1

0.1

0.2

0.1

0.9

0.3

0.3

0.4

0.7

89

τ-Cadinol#

1633

1629

2156

90

τ-Muurolol

1633

1635

2151

91

α-Cadinol#

1643

1637

2225

92

β-Eudesmol

1641

1638

2229

tr

0.4

0.4

0.3

0.2

0.4

0.6

0.8

0.5

0.1

1.0

93

α-Bisabolol

1673

1664

2190

0.6

0.2

0.1

0.6

0.4

0.2

0.3

0.2

0.2

0.9

1.5

94

Chamazulene

1719

1707

2396

0.2

tr

tr

0.1

0.1

tr

tr

tr

0.1

0.1

tr

Monoterpene hydrocarbons

11.1

12.6

13.5

9.9

14.7

10.9

9.9

11.6

1.8

4.7

18.7

Sesquitepene hydrocarbons

5.1

4.9

3.2

2.9

3.3

4.2

12.6

3.3

11.1

3.3

7.0

Oxygenated monoterpenes

71.6

74.7

67.9

73.0

58.6

70.2

60.8

67.5

62.8

66.2

55.0

Oxygenated sesquiterpenes

10.0

6.3

7.5

7.8

17.4

11.9

8.5

7.8

17.4

19.8

14.8

Others

0.4

0.3

0.2

0.2

0.2

0.5

0.2

0.1

0.3

0.2

0.5

Total identified

98.0

98.7

92.2

93.8

94.1

97.6

91.8

90.3

93.2

94.1

95.8

Yields (w/w vs fresh material)

0.89

0.85

0.90

0.79

0.81

0.13

0.18

0.20

0.15

0.21

 

Components

lRIa

RIa

RIp

Sardinian samples

Coll. Oil

Flowers

Leafy stems

F

G

H

I

F

G

H

I

  

1

Nonene

886

880

930

  

2

Santolinatriene

909

901

1018

1.4

5.0

1.3

0.9

1.9

0.6

0.7

  

3

Tricyclene

927

921

1005

 

tr

0.1

0.1

0.1

  

4

α-Tthujene

932

921

1008

tr

0.1

tr

0.1

  

5

α-Pinene

936

931

1008

0.6

0.8

0.1

1.2

0.7

0.4

0.1

1.4

0.4

  

6

Camphene

950

944

1045

0.7

1.1

0.1

1.6

0.4

0.6

0.1

1.5

0.2

  

7

Sabinene

973

965

1106

1.3

1.1

0.3

2.3

1.5

0.9

0.1

2.8

1.1

  

8

β-Pinene

978

971

1090

1.0

0.7

0.1

2.7

0.2

0.6

0.1

3.4

0.3

  

9

Myrcene

987

975

1143

tr

0.1

0.1

0.1

0.1

tr

0.1

0.1

0.1

  

10

Yomogi alcohol

991

984

1390

0.9

0.5

0.2

1.0

0.6

0.5

0.2

1.8

1.7

  

11

α-Phellandrene

1002

998

1142

0.1

0.5

tr

tr

0.1

0.2

tr

tr

tr

  

12

α-Terpinene

1013

1010

1156

0.3

0.5

0.1

0.4

0.5

0.2

0.1

0.4

0.3

  

13

p-Cymene

1015

1013

1245

1.3

1.0

0.7

1.1

1.2

1.5

0.8

1.7

1.2

  

14

Limonene

1023

1021

1180

0.3

2.0

0.1

2.8

1.4

1.0

3.3

0.1

  

15

β-Phellandrene#

1023

1021

1186

1.7

2.0

1.0

1.1

3.0

1.1

0.4

  

16

1,8-Cineole

1023

1021

1196

2.0

2.4

0.8

5.4

2.9

2.9

0.4

6.5

4.4

  

17

Santolina alcohol

1029

1021

1385

9.4

14.9

11.6

8.4

3.2

13.6

9.5

4.3

4.1

  

18

cis-β-Ocimene#

1029

1030

1215

  

19

trans-β-Ocimene#

1041

1040

1239

 

tr

tr

tr

tr

tr

tr

tr

tr

  

20

Artemisia ketone

1044

1046

1325

5.0

0.8

0.70

11.4

3.1

1.1

0.6

13.4

2.4

  

21

γ-Terpinene

1051

1050

1220

0.5

0.7

0.2

1.1

0.8

0.4

1.2

0.5

  

22

Isolyratol#

1052

1053

1524

tr

0.4

0.1

0.2

  

23

trans-Sabinene hydrate

1053

1054

1440

tr

0.1

0.1

0.2

0.2

0.1

0.2

  

24

Artemisia alcohol

1073

1069

1478

0.6

0.4

0.1

0.4

0.4

0.5

0.2

0.8

0.9

  

25

Terpinolene

1082

1079

1258

0.1

0.1

0.1

0.2

0.2

0.1

0.2

0.1

  

26

Linalool

1085

1084

1527

1.5

0.8

7.3

0.2

1.7

2.1

0.6

0.5

  

27

Filifolone

1086

1085

1419

0.1

0.8

3.7

2.7

0.5

0.8

  

28

α-Thujone

1089

1087

1402

1.2

2.3

8.9

0.3

0.2

1.6

3.4

0.4

1.0

  

29

β-Thujone

1103

1093

1422

1.3

1.0

1.6

0.8

2.2

0.8

0.5

1.2

  

30

Chrysanthenone

1110

1101

1480

0.8

0.2

4.0

0.2

1.5

0.3

6.5

tr

1.2

  

31

cis-p-Menth-2-en-1ol

1108

1108

1533

1.8

3.8

0.6

0.2

1.2

0.3

0.9

0.4

2.4

  

32

Camphor

1123

1123

1490

3.2

1.2

9.0

10.2

0.7

1.3

6.9

9.8

0.2

  

33

trans-Sabinol

1120

1123

1664

14.7

12.4

5.9

7.6

20.4

13.2

3.1

4.0

11.8

  

34

Pinocarvone

1137

1140

1558

0.2

0.2

0.4

0.5

0.1

0.2

0.4

0.5

0.1

  

35

cis-Chrysanthenol

1147

1146

1716

0.4

0.2

1.5

0.4

0.8

tr

0.4

tr

0.2

  

36

Borneol

1150

1153

1664

3.7

4.0

4.6

3.8

2.0

4.7

9.5

4.4

1.9

  

37

Artemisyl acetate

1153

1155

1390

0.2

0.6

tr

tr

0.5

0.6

  

38

Terpinen-4-ol

1164

1164

1570

2.3

1.4

2.3

2.5

2.4

1.9

1.9

3.7

2.7

  

39

Myrtenal

1172

1173

1612

tr

0.1

1.1

0.4

0.1

  

40

α-Terpineol

1176

1179

1664

0.8

0.5

0.8

0.2

0.8

1.2

1.3

1.3

  

41

Myrtenol

1178

1176

1772

tr

0.2

0.1

tr

0.1

  

42

cis-Piperitol#

1181

1181

1641

0.4

0.9

0.5

0.1

0.9

0.4

0.6

  

43

trans-Piperitol

1193

1191

1709

0.7

1.3

0.3

0.4

1.5

0.5

0.1

1.0

  

44

Piperitone

1226

1229

1696

2.9

5.0

0.6

0.6

2.6

3.8

1.5

0.8

3.7

  

45

cis-Chrysanthenyl acetate

1253

1244

1544

0.6

0.4

0.1

1.6

0.6

0.1

1.1

  

46

Bornyl acetate

1270

1271

1548

2.3

3.8

4.1

2.6

0.7

4.7

11.4

5.0

1.3

  

47

Thymol

1267

1272

2145

tr

0.1

0.7

tr

0.2

  

48

trans-Sabinyl acetate

1278

1274

1622

18.3

10.3

3.3

8.9

28.1

13.0

4.2

6.2

26.6

  

49

cis-Carvyl acetate

1318

1315

1710

tr

0.1

0.1

0.1

tr

0.1

0.1

  

50

Eugenol

1331

1329

2159

0.1

0.1

0.1

0.2

0.1

0.1

0.1

0.1

  

51

trans-Carvyl acetate

1345

1340

1749

  

52

α-Cubebene#

1355

1346

1435

  

53

α-Longipinene#

1360

1350

1457

  

54

α-Ylangene#

1376

1369

1458

tr

tr

tr

tr

tr

tr

tr

tr

tr

  

55

α-Copaene

1379

1374

1470

0.1

tr

0.1

tr

tr

tr

tr

tr

tr

  

56

β-Bourbonene#

1379

1386

1506

tr

tr

tr

tr

tr

tr

tr

tr

tr

  

57

β-Elemene

1389

1385

1570

  

58

α-Gurjunene

1413

1408

1507

  

59

β-Caryophyllene

1421

1424

1578

0.2

0.2

0.3

0.2

tr

0.4

0.4

0.5

0.3

  

60

β-Copaene#

1430

1425

1565

  

61

Aromadendrene

1443

1436

1597

  

62

trans-β-Farnesene

1446

1448

1629

tr

0.1

0.1

0.1

  

63

Alloaromadendrene

1462

1462

1620

tr

0.2

0.4

0.1

0.2

0.3

0.4

0.4

  

64

α-Curcumene

1473

1473

1750

0.2

0.1

0.3

0.1

0.2

0.1

0.1

0.2

  

65

γ-Curcumene#

1472

1474

1669

  

66

γ-Muurolene

1474

1474

1664

tr

tr

tr

tr

tr

tr

tr

tr

tr

  

67

Germacrene-D

1479

1480

1682

2.3

1.0

2.4

1.2

0.8

5.5

0.9

7.2

7.2

  

68

β-Selinene#

1486

1483

1695

  

69

Zingiberene

1489

1489

1698

  

70

Bicyclo germacrene

1494

1492

1710

tr

0.2

0.1

0.1

0.2

0.5

0.3

0.4

0.6

  

71

Ledene

1491

1492

1669

  

72

α-Muurolene

1496

1494

1699

  

73

trans,trans-α-Farnesene#

1498

1499

1730

  

74

γ-Cadinene

1507

1507

1729

  

75

Calamenene

1517

1512

1802

  

76

δ-Cadinene

1520

1515

1729

tr

0.2

0.5

0.2

0.1

0.4

0.4

0.3

0.4

  

77

Cadina-1,4-diene#

1523

1523

1763

  

78

α-Calacorene

1527

1531

1880

tr

0.3

0.4

0.2

0.2

0.2

0.4

0.2

  

79

trans-Nerolidol

1533

1546

2031

tr

tr

0.5

0.1

tr

  

80

Elemol

1541

1536

2060

  

81

β-Calacorene#

1566

1548

1913

  

82

Spathulenol

1572

1560

2101

tr

tr

0.3

0.2

  

83

Palustrol

1569

1563

1903

  

84

Caryophyllene oxide

1578

1570

1960

0.2

0.8

0.5

0.6

0.1

0.4

2.0

0.6

0.3

  

85

Viridiflorol

1592

1580

2039

2.6

2.8

7.5

1.7

3.1

2.9

12.6

1.7

3.4

  

86

Ledol

1600

1593

1983

0.2

0.5

0.6

0.4

0.2

  

87

epi-Cubenol

1623

1614

2032

0.1

1.2

0.1

0.3

0.1

  

88

γ-Eudesmol

1618

1622

2170

  

89

τ-Cadinol#

1633

1629

2156

0.5

0.9

0.3

0.5

0.5

0.5

0.5

  

90

τ-Muurolol

1633

1635

2151

0.1

0.7

0.3

0.2

0.1

0.5

0.5

  

91

α-Cadinol#

1643

1637

2225

0.1

0.4

0.2

0.1

0.5

0.5

1.7

0.2

0.3

  

92

β-Eudesmol

1641

1638

2229

0.5

0.5

0.5

0.1

1.1

0.6

0.5

  

93

α-Bisabolol

1673

1664

2190

0.2

0.6

0.5

0.5

0.1

1.1

0.2

0.1

  

94

Chamazulene

1719

1707

2396

  

Monoterpene hydrocarbons

9.5

15.7

2.0

16.1

9.3

10.9

1.5

17.9

5.7

  

Sesquitepene hydrocarbons

3.2

2.4

4.8

2.4

1.9

7.7

3.1

8.7

9.5

  

Oxygenated monoterpenes

75.6

69.5

71.7

70.9

81.0

68.4

66.6

65.1

73.4

  

Oxygenated sesquiterpenes

3.7

4.8

13.0

4.4

4.2

3.8

18.8

3.4

4.3

  

Others

0.1

0.1

0.1

0.2

0.1

0.1

0.1

0.1

  

Total identified

92.1

92.4

91.5

94.0

96.5

90.8

90.1

95.2

93.0

  

Yields (w/w vs fresh material)

0.85

0.76

0.89

0.82

0.21

0.18

0.16

0.19

  

Order of elution and percentages of individual components are given on Rtx-1 column. RIa and RIp: retention index on Rtx-1 apolar column and Rtx-Wax polar column, respectively. lRIa: All retention indices on apolar column were reported from literature data [32, 35], except for nonene and β-calacorene [33, 34]. The components with (#) were identified for the first time in the A. ligustica oil. tr: trace (<0.05). Coll. Oil: mixture of samples. Localities of sampling: A = St Georges, B = Ghisoni, C = Cervioni, D = Propriano, E = Corti, F = M. Spada, G = St Ana, H = M. Limbara, I = Baddeurbara

2.2 Oil Fractionation

The collective oil (2.8 g) obtained by the mixture of all Corsican sample oils from aerial parts (flowers and leafy stems) was submitted to flash chromatography (FC, silica gel 200–500 μm, elution with pentane (PE), then diethyl ether (Et2O)) and was separated into two fractions: F1 apolar (0.45 g) and F2 polar (2.19 g). As above, the collective oil (3.5 g) obtained by the mixture of all Sardinian sample oils from aerial parts (flowers and leafy stems) was submitted to flash chromatography and was separated into two fractions: F1 apolar (0.59 g) and F2 polar (2.82 g).

2.3 HS-SPME Conditions

The fresh aerial parts (mixture of whole flowers and leafy stems) and the single organs (flowers and leafy stems separately) of A. ligustica were cut roughly with scissors (1–2 cm long) before subjection to headspace-solid phase microextraction (HS-SPME). The SPME device (Supelco) coated with divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS, 30 μm) was used for extraction of the plant volatiles. Optimization of conditions was carried out using fresh aerial parts of the plant (700 mg in a 20 mL vial) and based on the sum of total peak areas measured on GC-FID. Equilibration time was fixed at 60 min. Temperature and extraction time were selected after nine experiments combining three temperatures (50, 70, 90 °C) and three times (15, 30 and 45 min). After sampling, SPME fibre was inserted into the GC and GC-MS injection ports for desorption of volatile components (5 min), both using the splitless injection mode. Before sampling, each fibre was reconditioned for 5 min in the GC injection port at 260 °C. HS-SPME and subsequent analyses were performed in triplicate. The coefficient of variation (9.6% < CV < 13.4%) calculated on the basis of total area obtained from the FID-signal for the samples indicated that the HS-SPME method produced reliable results. In the same way, the CV of the major compounds was always less than 15%.

2.4 Gas Chromatography

GC analyses were carried out using a Perkin Elmer Autosystem GC apparatus (Walhton, MA, USA) equipped with a single injector and two flame ionization detectors (FID). The apparatus was used for simultaneous sampling to two fused-silica capillary columns (60 m × 0.22 mm, film thickness 0.25 μm) with different stationary phases: Rtx-1 (polydimethylsiloxane) and Rtx-Wax (polyethylene glycol). Temperature program: 60 to 230 °C at 2 °C min−1 and then held isothermal 230 °C (30 min). Carrier gas: helium (1 mL min−1). Injector and detector temperatures were held at 280 °C. Split injection was conducted with a ratio split of 1:80. Injected volume: 0.1 μL. For HS-SPME-GC analysis, only Rtx-1 (polydimethylsiloxane) column was used and volatile components were desorbed in a GC injector with a SPME inlet liner (0.75 mm. I.D., Supelco).

2.5 Gas Chromatography-Mass Spectrometry

The oils and the fractions obtained by CC were investigated using a Perkin Elmer TurboMass quadrupole detector, directly coupled to a Perkin Elmer Autosystem XL equipped with two fused-silica capillary columns (60 m × 0.22 mm, film thickness 0.25 μm), Rtx-1 (polydimethylsiloxane) and Rtx-Wax (polyethylene glycol). Other GC conditions were the same as described above. Ion source temperature: 150 °C; energy ionization: 70 eV; electron ionization mass spectra were acquired with a mass range of 35–350 Da. Oil injected volume: 0.1 μL, fractions injected volume: 0.2 μL. The volatile fractions sampling by HS-SPME were analyzed only on a Rtx-1 capillary column and volatile components were desorbed in a GC injector with a SPME inlet liner (0.75 mm. I.D., Supelco).

2.6 Component Identification

Identification of the components was based (i) on the comparison of their GC retention indices (RI) on non polar and polar columns, determined relative to the retention time of a series of n-alkanes with linear interpolation, with those of authentic compounds or literature data [32, 33, 34, 35, 38]; and (ii) on computer matching with commercial mass spectral libraries [35, 36, 37, 38] and comparison of spectra with those of our personal library. Relative amounts of individual components were calculated on the basis of their GC peak areas on the two capillary Rtx-1 and Rtx-Wax columns, without FID response factor correction.

3 Results and Discussion

3.1 Essential Oil Compositions

The analysis of Corsican and Sardinian A. ligustica collective oils and fractions obtained after flash chromatography allowed the identification of 76 and 72 components, respectively, accounting for more than 90% of the oils. In total, 37 monoterpenes, 37 sesquiterpenes and two non-terpenic compounds were identified in the Corsican oil and 49 monoterpenes, 22 sesquiterpenes and one non-terpenic compound were identified in the Sardinian oil. Their retention indices and their relative percentages are presented in Table 1. Among the 94 individual compounds identified, 90 were performed by comparing their EI-mass spectra and their retention indices with those of our own library. The majority of these compounds were commercial standard components and the few others, were previously identified at the large amount in essential oils or fractions obtained by CC, by comparison with literature spectral data and retention indices and ensured by 13C-NMR. In the present study, only trans-sabinol, cis-chrysanthenyl acetate, trans-sabinyl acetate and cadina-1,4-diene, were identified by comparison of their EI-mass spectra and their retention indices with those of commercial libraries and/or literature data.

These oils were characterized by a large variety of regular terpenic skeletons: acyclic, menthane, bicyclo[3.1.1]heptane, bicyclo[3.1.0]hexane and bicyclo[2.2.1]heptane, farnesane, germacrane, bicyclogermacrane, caryophyllane, cadinane, muurolane, eudesmane, copaane and guaiane but also by two irregular terpenic of non-head-to-tail isoprenoid skeletons: santolinane and artemisane. Camphor (20.8%), santolina alcohol (15.4%), viridiflorol (7.9%), borneol (7.8%) and bornyl acetate (5.9%) were the major components of the Corsican oils, while trans-sabinyl acetate (18.3%), trans-sabinol (14.7%), santolina alcohol (9.4%) and artemisia ketone (5.0%) were identified as main components in the Sardinian oils.

As reported in the literature [13, 14], Corsican and Sardinian A. ligustica oils exhibited quantitative differences on their chemical compositions. These island oils differed also from an another Mediterranean island oil from Sicilia, in which terpinen-4-ol (19%), carvone (9%) and γ-terpinene (7%) were identified as major compounds. The Corsican and Sardinian oils investigated in this study were drastically different from the sample oil obtained from continental Italy in which artemisia acetate (44%), 2,7-dimethyl-4,6-octadien-2-ol (16%) and linalool (10%) were identified [16].

All flower and leafy stem oils were dominated by the high content of monoterpene compounds, especially oxygenated monoterpenes which accounted for always more than 55%. Among them, six irregular monoterpenes which accounted for 8.7–22.6% in the Sardinian oils and 16.9–49.8% in the Corsican oils were identified: santolinatriene, santolina alcohol and isolyratol (santolinyl group), yomogi alcohol, artemisia alcohol and artemisia acetate (artemisyl group). A relative similarity was noted for these oils showing a homogenous qualitative composition, in contrast to a strong percentage variability depending on the origin of the samples. The major components of Corsican oils were santolina alcohol (2.9–30.1%), camphor (3.9–26.2%), artemisia ketone (3.2–17.7%), trans-sabinyl acetate (0.2–16.6%), viridiflorol (4.4–15.0%) and linalool (0.2–11.6%). The Sardinian oils exhibited trans-sabinyl acetate (3.3–28.1%), trans-sabinol (3.1–20.4%), santolina alcohol (3.2–14.9%), artemisia ketone (0.6–13.4%), viridiflorol (1.7–12.6%), bornyl acetate (1.3–11.4%) and camphor (0.2–10.2%) as main components. Moreover, the Corsican and Sardinian flower oils differed from these reported from Greece in which linalool was predominant (78%) [17].

Finally, this study allowed the identification of 22 and 19 compounds not yet reported in the Corsican and Sardinian A. ligustica oils, respectively. Among them, 17 were identified for the first time in A. ligustica oils: β-phellandrene, trans-β-ocimene, cis-β-ocimene, isolyratol, cis-piperitol, α-cubebene, α-longipinene, α-ylangene, β-bourbonene, β-copaene, γ-curcumene, β-selinene, trans-trans-α-farnesene, cadina-1,4-diene, β-calacorene, τ-cadinol, and α-cadinol.

3.2 HS-SPME Analysis of Volatiles Constituents

The A. ligustica volatile compounds of whole aerial parts, flowers and leafy stems of the sample from Corti (Corsica) were investigated using HS-SPME under optimized parameters. The optimization of HS-SPME sampling parameters was carried out using fresh aerial parts based on the sum of total peak areas obtained by GC-FID. The maximum sum of total peak area was acquired for a temperature of 70 °C, an equilibrium time of 60 min and an extraction time of 30 min (Table 2). The sum of total peak area increased according to the raise of the temperature. The GC-RI and GC-MS analysis allowed the identification of 57 components including 13 monoterpene hydrocarbons, 16 sesquiterpene hydrocarbons, 20 oxygenated monoterpenes, six oxygenated sesquiterpenes and two non terpenic compounds (Table 3).
Table 2

Total peak areas (FID. 105 mV) of volatile fraction of Achillea ligustica sampling from different HS-SPME parameters (temperatures and extraction times)

Times

Temperatures (°C)

50

70

90

15 min

175

198

211

30 min

201

274

246

45 min

231

261

204

The equilibrium time was fixed at 60 min

Table 3

Volatile components extracted by HS-SPME (% SPME) and hydrodistillation (% HD) of Achillea ligustica from Corti (Corsica)

 

Components

lRIa

RIa

RIp

% SPME

% HD

Aerial Parts

Flowers

Leafy stems

Aerial Parts

Flowers

Leafy stems

1

Nonene

886

880

930

0.2 ± 0.01

0.2 ± 0.01

0.4 ± 0.02

0.1

0.1

0.4

2

Santolinatriene

909

901

1018

1.0 ± 0.11

1.8 ± 0.11

1.5

0.7

2.2

3

Tricyclene

927

921

1005

0.4 ± 0.01

0.6 ± 0.02

0.1

0.1

0.2

4

α-Thujene

932

921

1008

0.1 ± 0.01

0.1 ± 0.02

0.1

0.1

0.1

5

α-Pinene

936

931

1008

1.7 ± 0.22

3.4 ± 0.31

1.5 ± 0.20

1.7

0.9

2.4

6

Camphene

950

944

1045

4.8 ± 0.59

9.0 ± 1.80

3.0 ± 0.39

3.1

1.4

3.3

7

Sabinene

973

965

1106

0.8 ± 0.11

1.1 ± 0.01

0.6 ± 0.05

1.1

1.6

1.6

8

β-Pinene

978

971

1090

3.6 ± 0.51

4.3 ± 0.81

2.6 ± 0.56

3.9

3.7

5.2

9

Myrcene

987

975

1143

0.1 ± 0.01

0.2 ± 0.01

0.1

0.2

0.1

10

Yomogi alcohol

991

984

1390

0.4 ± 0.01

0.8 ± 0.08

0.7

0.3

0.5

11

α-Phellandrene

1002

998

1142

tr

0.1 ± 0.01

tr

tr

0.1

12

α-Terpinene

1013

1010

1156

0.8 ± 0.10

1.0 ± 0.08

0.3 ± 0.02

0.8

0.5

0.9

13

p-Cymene

1015

1013

1245

0.7 ± 0.08

1.1 ± 0.09

0.2 ± 0.02

0.9

0.3

0.5

14

Limonene

1023

1021

1180

tr

tr

tr

16

1,8-Cineole

1023

1021

1196

1.1 ± 0.15

1.3 ± 0.18

0.3 ± 0.01

2.8

3.4

0.4

17

Santolina alcohol

1029

1021

1385

3.8 ± 0.48

0.5 ± 0.10

9.4 ± 1.26

9.1

3.8

10.1

18

cis-β-Ocimene

1029

1030

1215

tr

tr

tr

19

trans-β-Ocimene

1041

1040

1239

tr

tr

tr

20

Artemisia ketone

1044

1046

1325

20.4 ± 2.89

0.3 ± 0.05

26.7 ± 3.55

7.5

5.9

3.2

21

γ-Terpinene

1051

1050

1220

1.1 ± 0.12

2.3 ± 0.50

0.8 ± 0.02

1.5

1.0

1.7

23

trans-Sabinene hydrate

1053

1054

1440

1.3 ± 0.09

0.5 ± 0.07

4.1 ± 0.51

1.6

0.1

4.2

24

Artemisia alcohol

1073

1069

1478

0.4 ± 0.06

1.2 ± 0.06

0.9

0.2

0.6

25

Terpinolene

1082

1079

1258

0.3 ± 0.01

0.5 ± 0.05

0.2 ± 0.01

0.3

0.2

0.3

26

Linalool

1086

1084

1527

0.9 ± 0.10

1.1 ± 0.20

0.3 ± 0.02

2.9

11.6

1.1

29

β-Thujone

1103

1093

1422

0.6

1.0

0.4

32

Camphor

1123

1123

1490

22.8 ± 2.12

29.8 ± 2.50

14.2 ± 2.10

17.4

17.2

17.0

33

trans-Sabinol

1120

1123

1664

1.5 ± 0.15

1.1 ± 0.12

0.9 ± 0.05

4.9

7.4

1.5

34

Pinocarvone

1137

1140

1558

0.5 ± 0.06

0.4 ± 0.05

0.2 ± 0.01

0.2

0.2

0.4

36

Borneol

1150

1153

1664

5.0 ± 0.53

6.7 ± 0.38

0.5 ± 0.02

3.5

2.6

1.3

37

Artemisyl acetate

1153

1155

1390

0.8 ± 0.11

6.0 ± 0.71

0.1

tr

tr

38

Terpinen-4-ol

1164

1164

1570

3.7 ± 0.31

2.6 ± 0.23

0.1 ± 0.01

2.6

2.7

3.2

39

Myrtenal

1172

1173

1612

0.1 ± 0.01

0.4 ± 0.02

0.9 ± 0.05

0.1

0.1

0.2

40

α-Terpineol

1176

1179

1664

0.6 ± 0.10

0.2 ± 0.01

0.3 ± 0.01

0.8

0.7

0.6

45

cis-Chrysanthenyl acetate

1253

1244

1544

1.1 ± 0.08

2.2 ± 0.15

0.7 ± 0.02

0.5

0.3

0.3

46

Bornyl acetate

1270

1271

1548

2.1 ± 0.05

3.2 ± 0.30

2.1 ± 0.04

2.7

2.1

2.0

48

trans-Sabinyl acetate

1278

1274

1622

3.0 ± 0.28

5.5 ± 0.56

1.6 ± 0.21

6.2

10.2

7.6

49

cis-Carvyl acetate

1318

1315

1710

1.0 ± 0.08

1.7 ± 0.17

0.9 ± 0.02

0.2

0.2

0.2

50

Eugenol

1331

1329

2159

0.3 ± 0.01

tr

0.3 ± 0.02

0.3

0.4

0.1

51

trans-Carvyl acetate

1345

1340

1749

0.5 ± 0.01

1.0 ± 0.05

0.4 ± 0.01

tr

0.1

0.1

52

α-Cubebene

1355

1346

1435

tr

0.1 ± 0.02

tr

tr

tr

tr

53

α-Longipinene

1360

1350

1457

tr

tr

tr

54

α-Ylangene

1376

1369

1458

tr

0.1 ± 0.01

tr

0.1

tr

tr

55

α-Copaene

1379

1374

1470

0.1 ± 0.01

0.4 ± 0.02

0.1 ± 0.01

0.2

tr

tr

57

β-Elemene

1389

1385

1570

0.1 ± 0.01

0.3 ± 0.02

tr

0.1

tr

tr

58

α-Gurjunene

1413

1408

1507

0.1 ± 0.01

0.1 ± 0.01

0.1

tr

tr

59

β-Caryophyllene

1421

1424

1578

0.2

tr

0.4

60

β-Copaene

1430

1425

1565

tr

0.1 ± 0.01

0.1

tr

tr

61

Aromadendrene

1443

1436

1597

tr

0.2 ± 0.01

tr

tr

tr

62

trans-β-Farnesene

1446

1448

1629

0.1

0.1

0.1

63

Alloaromadendrene

1462

1462

1620

1.1 ± 0.12

1.6 ± 0.10

1.4 ± 0.20

0.5

0.4

0.6

64

α-Curcumene

1473

1473

1750

0.1

tr

tr

65

γ-Curcumene

1472

1474

1669

0.4 ± 0.02

0.4 ± 0.11

tr

tr

tr

67

Germacrene-D

1479

1480

1682

1.9 ± 0.21

0.3 ± 0.01

1.6 ± 0.08

2.1

1.5

3.2

68

β-Selinene

1486

1483

1695

tr

tr

tr

69

Zingiberene

1489

1489

1698

tr

tr

tr

70

Bicyclogermacrene

1494

1492

1710

0.3

0.2

0.3

71

Ledene

1491

1492

1669

0.1

tr

tr

72

α-Muurolene

1496

1494

1699

0.7 ± 0.10

0.2 ± 0.01

1.9 ± 0.28

tr

tr

tr

73

trans,trans-α-Farnesene

1498

1499

1730

0.2

0.3

0.3

74

γ-Cadinene

1507

1507

1729

0.1 ± 0.01

0.4 ± 0.05

0.1 ± 0.02

tr

tr

tr

75

Calamenene

1517

1512

1802

0.1 ± 0.01

0.1 ± 0.02

tr

tr

tr

76

δ-Cadinene

1520

1515

1729

3.6 ± 0.51

5.8 ± 0.85

3.2 ± 0.43

0.8

0.6

0.9

77

Cadina-1,4-diene

1523

1523

1763

tr

tr

tr

78

α-Calacorene

1527

1531

1880

0.1 ± 0.01

0.1 ± 0.01

0.1 ± 0.02

0.1

0.1

0.2

80

Elemol

1541

1536

2060

0.5

0.6

0.4

81

β-Calacorene

1566

1548

1913

tr

tr

tr

tr

tr

tr

82

Spathulenol

1572

1560

2101

0.3

0.2

0.3

83

Palustrol

1569

1563

1903

0.2 ± 0.01

0.2 ± 0.01

0.2 ± 0.01

tr

tr

tr

84

Caryophyllene oxide

1578

1570

1960

0.1

0.1

0.2

85

Viridiflorol

1592

1580

2039

2.7 ± 0.31

3.0 ± 0.45

2.3 ± 0.11

7.2

8.2

9.5

86

Ledol

1600

1593

1983

0.2 ± 0.01

0.3 ± 0.01

0.2 ± 0.01

0.5

0.6

0.6

87

epi-Cubenol

1623

1614

2032

1.7 ± 0.22

0.7 ± 0.05

0.1 ± 0.01

0.5

0.6

0.5

88

γ-Eudesmol

1618

1622

2170

0.8

0.9

0.7

92

β-Eudesmol

1641

1638

2229

tr

tr

2.1 ± 0.18

0.9

0.4

1.0

93

α-Bisabolol

1673

1664

2190

tr

tr

tr

1.1

0.2

1.5

94

Chamazulene

1719

1707

2396

tr

tr

tr

Monoterpene hydrocarbons

15.5

23.6

11.1

15.3

10.9

18.7

Sesquitepene hydrocarbons

8.6

10.3

8.6

5.7

4.2

7.0

Oxygenated monoterpenes

66.5

51.9

69.1

65.4

70.2

55.0

Oxygenated sesquiterpenes

4.9

4.3

5.0

12.0

11.9

14.8

Others

0.5

0.3

0.7

0.4

0.5

0.5

Total identified

95.9

90.3

94.5

98.6

97.6

95.8

Yields (w/w vs fresh material)

0.58

0.81

0.21

Total area/104 (expressed in arbitrary units)

7.56 ± 1.02

9.15 ± 0.88

1.42 ± 0.12

Order of elution and percentages (means of three experiments) of individual components are given on Rtx-1 column. RIa and RIp: retention index on Rtx-1 apolar column and Rtx-Wax polar column, respectively. lRIa: Retention indices on apolar column reported from literature [32, 35] except for nonene and β-calacorene [33, 34]. tr: trace (<0.05)

The HS-fractions obtained from fresh plant materials (whole aerial parts, flowers and leafy stems) were rather qualitatively similar but differed from the percentages of their major components (Fig. 1). We noted that the volatile constituents were more abundant in the flowers than in the other parts of the plant. In the HS-fractions obtained from aerial parts and flowers, camphor (22.8 and 29.8%, respectively) was the major component, followed by artemisia ketone (20.4%) and santolina alcohol (3.8%) in whole aerial parts compared to camphene (9.0%) and trans-sabinyl acetate (5.5%) in flowers. Conversely, the leafy stem HS-fraction was characterized by the presence of artemisia ketone (26.7%), camphor (14.2%) santolina alcohol (9.4%) as main components. It is noticeable that the irregular monoterpenes amounted for 26.8% and 45.9% in the HS-fractions obtained from whole aerial parts and leafy stems and only 1.0% in the HS-fraction from flowers.
Fig. 1

GC-chromatograms of volatile components obtained by HS-SPME (I) and hydrodistillation (II) from leafy stems (a), flowers (b) and total aerial parts (c) of A. ligustica from Corti (Corsica)

Concerning the HD-oils from the same plant material, camphor was always the major component (17.0–17.4%) followed by santolina alcohol (9.1%) and artemisia ketone (7.5%) in whole aerial parts, linalool (11.6%) and trans-sabinyl acetate (10.2%) in flowers oil and santolina alcohol (10.1%) and viridiflorol (9.5%) in leafy stem oil. Quantitative differences between HS and HD-analysis were observed for oxygenated sesquiterpenes which were always present in superior amounts in the essential oils. It was difficult to establish a direct correlation between HS and HD-extraction techniques because the first was controlled by an equilibrium step and the latter was based on the quasi-total extraction of plant volatiles. Because the experiments were carried out for the optimization of SPME extraction parameters, it appears that the extraction temperature was the most important parameter for the plant headspace study. The distribution constants of each compound were temperature-dependent: at medium temperature (50 °C) and at high temperature (90 °C), extractions of hydrocarbons monoterpenes and oxygenated sesquiterpenes were improved. The optimal temperature (70 °C) used for the HS-extraction was an analytical compromise based on the maximal amount of volatiles extracted. Concerning the comparison of both techniques in terms of isolation time, HS-SPME was clearly fast (90 min), while 300 min were required for hydrodistillation. In the same way, the amount of plant material used for the headspace analysis was smaller (0.7 g), while the production of A. ligustica oil by hydrodistillation needed the use of 110–190 g of plant material. This might be one of the major reasons which explain the difference of chemical HS and HD data.

This study demonstrates that the HS-SPME analysis allowed to obtain rapidly the fingerprints of plant headspace. Headspace extraction was performed using a smaller amount of plant material and so HS-SPME is well suited for the analysis of volatile compounds of the whole plant and/or separated organs which yielded low or no essential oil. Moreover, the sampling of the plant volatile components by HS-SPME at medium temperatures (70 °C), ensure the extraction of a volatile fraction which is representative of the essential oil. Only quantitative differences of some components can be observed in both aromatic profiles, while qualitatively both aromatic mixtures are rather similar. For these reasons, HS-SPME can be applied to routine control analysis of aromatic and medicinal plants.

Notes

Acknowledgments

The authors are indebted to the Collectivité Territoriale de Corse and the European Community for partial financial support (PIC INTERREG IIIA).

References

  1. 1.
    Gamisans J, Jeanmonod D (1998) In: Asteraceae-I. Compléments au Prodrome de la Flore Corse. Conservatoire et Jardin botaniques, GenèveGoogle Scholar
  2. 2.
    Pignatti S (1982) In: Flora d’Italia; Edagricole, BolognaGoogle Scholar
  3. 3.
    Glasl S, Mucaji P, Werner I, Presser A, Jurenitschi J (2002) Z Naturforsch 57c:978–982Google Scholar
  4. 4.
    Atzei AD (2003) In: Le Piante nella Tradizione Popolare della Sardegna; Carlo Delfino, SassariGoogle Scholar
  5. 5.
    Simonpoli P (1993) In: Arburi, Arbe, Arbigliule, Savoirs Populaires sur les Plantes de Corse; Parc Naturel Régional de la Corse, AjaccioGoogle Scholar
  6. 6.
    Bruno M, Herz W (1988) Phytochemistry 27:1871–1872. doi:10.1016/0031-9422(88)80466-7 CrossRefGoogle Scholar
  7. 7.
    Tzakou O, Couladis M, Verykokidou E, Loukis A (1995) Biochem Syst Ecol 23:569–570. doi:10.1016/0305-1978(95)00044-U CrossRefGoogle Scholar
  8. 8.
    Ahmed A, Gati T, Hussein T-A, All A-T, Tzakou O-A, Couladis M, Mabry T-J, Toth G (2003) Tetrahedron 59:3729–3735. doi:10.1016/S0040-4020(03)00572-6 CrossRefGoogle Scholar
  9. 9.
    Grerger H, Zdero C, Bohlmann F (1984) Phytochemistry 23:1503–1505. doi:10.1016/S0031-9422(00)80494-X CrossRefGoogle Scholar
  10. 10.
    Boudjerda A, Zater H, Benayache S, Chalchat J-C, Gonzales-Platas J, Leon F, Brouard I, Bermejo J, Benayache F (2008) Biochem Syst Ecol 36:461–466. doi:10.1016/j.bse.2007.11.006 CrossRefGoogle Scholar
  11. 11.
    Conforti F, Loizzo M-R, Statti G-A, Menichini F (2005) Biol Pharm Bull 28:1791–1794. doi:10.1248/bpb.28.1791 CrossRefGoogle Scholar
  12. 12.
    Nemeth E (2005) J Essent Oil Res 17:501–512Google Scholar
  13. 13.
    Filippi J-J, Lanfranchi D-A, Prado S, Baldovini N, Meierhenrich U-J (2006) J Agric Food Chem 54:6308–6313. doi:10.1021/jf060752y CrossRefGoogle Scholar
  14. 14.
    Tuberoso C-I-G, Kowalczyk A, Coroneo V, Russo M-T, Dessi S, Cabras P (2005) J Agric Food Chem 53:10148–10153. doi:10.1021/jf0518913 CrossRefGoogle Scholar
  15. 15.
    Bader A, Panizzi L, Cioni PL, Flamini G (2007) Cent Eur J Biol 2:206–212. doi:10.2478/s11535-007-0020-3 CrossRefGoogle Scholar
  16. 16.
    Maffei M, Germano F, Doglia G, Chialva F (1993) J Essent Oil Res 5:61–70Google Scholar
  17. 17.
    Tzakou O, Loukis A, Verykokidou E, Roussis V (1995) J Essent Oil Res 7:549–550Google Scholar
  18. 18.
    Tholl D, Boland W, Hansel A, Loreto F, Rose USR, Schnitzler JP (2006) Plant J 45:540–560. doi:10.1111/j.1365-313X.2005.02612.x CrossRefGoogle Scholar
  19. 19.
    Bicchi C, Cordero C, Liberto E, Rubiolo P, Sgorbini B (2004) J Chromatogr A 1024:217–226. doi:10.1016/j.chroma.2003.10.009 CrossRefGoogle Scholar
  20. 20.
    Bicchi C, Cordero C, Liberto E, Sgorbini B, Rubiolo P (2007) J Chromatogr A 1152:138–149. doi:10.1016/j.chroma.2007.02.011 CrossRefGoogle Scholar
  21. 21.
    Flamini G, Cioni PL, Morelli I (2003) J Chromatogr A 998:229–233. doi:10.1016/S0021-9673(03)00641-1 CrossRefGoogle Scholar
  22. 22.
    Bertoli A, Pistelli L, Morelli I, Fraternale D, Giamperi L, Ricci D (2004) Flavour Fragr J 19:522–525. doi:10.1002/ffj.1382 CrossRefGoogle Scholar
  23. 23.
    Demirci B, Demirci F, Baser KHC (2005) Flavour Fragr J 20:395–398. doi:10.1002/ffj.1426 CrossRefGoogle Scholar
  24. 24.
    Pellati F, Benvenuti S, Yoshizaki F, Bertelli D, Rossi MC (2005) J Chromatogr A 1087:265–273. doi:10.1016/j.chroma.2005.01.060 CrossRefGoogle Scholar
  25. 25.
    Belliardo F, Bicchi C, Cordero C, Liberto E, Rubiolo P, Sgorbini B (2006) J Chromatogr Sci 44:416–429Google Scholar
  26. 26.
    Bicchi C, Drigo S, Rubiolo P (2000) J Chromatogr A 892:469–485. doi:10.1016/S0021-9673(00)00231-4 CrossRefGoogle Scholar
  27. 27.
    Paolini J, Nasica E, Desjobert J-M, Muselli A, Bernardini A-F, Costa J (2007) J Phytochem Anal 19:266–276. doi:10.1002/pca.1047 CrossRefGoogle Scholar
  28. 28.
    Rolhoff J, Skagen EB, Steen AH, Iversen T-H (2000) J Agric Food Chem 48:6205–6209. doi:10.1021/jf000720p CrossRefGoogle Scholar
  29. 29.
    Cornu A, Carnat A-P, Martin B, Coulon J-B, Lamaison J-L, Berdague J-L (2001) J Agric Food Chem 49:203–209. doi:10.1021/jf0008341 CrossRefGoogle Scholar
  30. 30.
    Jaenson TGT, Palsson K, Borg-Karlson A-K (2006) J Med Entomol 43:113–119. doi:10.1603/0022-2585(2006)043[0113:EOEAOO]2.0.CO;2 CrossRefGoogle Scholar
  31. 31.
    Conseil de l’Europe (1996) Pharmacopée Européenne, Maisonneuve S.A.: Sainte RuffineGoogle Scholar
  32. 32.
    Jennings W, Shibamoto T (1980) In: Qualitative analysis of flavour and fragrance volatiles by glass-capillary gas chromatography. Academic Press, New YorkGoogle Scholar
  33. 33.
    Liu K, Rossi P-G, Ferrari B, Berti L, Casanova J, Tomi F (2007) Phytochemistry 68:1698–1705. doi:10.1016/j.phytochem.2007.04.027 CrossRefGoogle Scholar
  34. 34.
    Kalemba D, Thiem B (2004) Flavour Fragr J 19:40–43. doi:10.1002/ffj.1271 CrossRefGoogle Scholar
  35. 35.
    MassFinder (version 3.0) (2006) In: Hochmunth D. Scientific Consulting, Germany Google Scholar
  36. 36.
    National Institute of Standards and Technology (1996) PC version 1.5a of the NIST/EPA/NIH Mass Spectral Library, Perkin Elmer CorporationGoogle Scholar
  37. 37.
    Mc Lafferty FW, Stauffer DB (1989) The Wiley/NBS registry of mass spectra data, vol 1–7. Wiley-Interscience publication, New-YorkGoogle Scholar
  38. 38.
    Adams RP (2001) Identification of essential oils by capillary gas chromatography/mass spectroscopy. Allured, Carol stream, ILGoogle Scholar

Copyright information

© Vieweg+Teubner | GWV Fachverlage GmbH 2009

Authors and Affiliations

  • Alain Muselli
    • 1
  • Marta Pau
    • 2
  • Jean-Marie Desjobert
    • 1
  • Marcia Foddai
    • 2
  • Marianna Usai
    • 2
  • Jean Costa
    • 2
  1. 1.Laboratoire de Chimie des Produits Naturels, CNRS UMR-6134 Université de CorseCortiFrance
  2. 2.Dipartimento di Scienze del FarmacoUniversita di SassariSassariItaly

Personalised recommendations