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Essential Oil Composition of Myrtle (Myrtus communis) Leaves

M. Mulas & R. A. M. Melis
a a a

Department of Economics and Tree Systems, University of Sassari, Sassari, Italy Available online: 11 Mar 2011

To cite this article: M. Mulas & R. A. M. Melis (2011): Essential Oil Composition of Myrtle (Myrtus communis) Leaves, Journal of Herbs, Spices & Medicinal Plants, 17:1, 21-34 To link to this article: http://dx.doi.org/10.1080/10496475.2011.556986

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Journal of Herbs, Spices & Medicinal Plants, 17:2134, 2011 Copyright Taylor & Francis Group, LLC ISSN: 1049-6475 print/1540-3580 online DOI: 10.1080/10496475.2011.556986

Essential Oil Composition of Myrtle (Myrtus communis) Leaves

M. MULAS and R. A. M. MELIS
Department of Economics and Tree Systems, University of Sassari, Sassari, Italy

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Nine cultivars of myrtle ( Myrtus communis) were analyzed for their leaf essential oil yield and composition during two seasons. The oil yield composition on leaf dry weight basis varied with the year of collection and the season ranging from 0.6 to 10.7 gkg1 in the different cultivars. The oils were analyzed by GCMS for compound identication and by GC for quantitative determinations by comparison with authentic standards. The main components of the essential oils were -pinene (366.8 to 816.8 gkg1 ); limonene (42.8 to 468.8 gkg1 ), 1,8-cineole (32.0 to 230.6 gkg1 ), linalool (0.8 to 110.3 gkg1 ), geranyl-acetate (12.4 to 99.2 gkg1 ). Other minor components were -terpineol (1.6 to 44.4 gkg1 ), terpinolene isomer 1 (0.0 to 42.6 gkg1 ), linalyl-acetate (0.0 to 36.3 gkg1 ), -humulene (0.0 to 19.6 gkg1 ), terpinolene-isomer 2 (0.0 to 14.9 gkg1 ), p-cymene (0.0 to 13.0 gkg1 ) and geraniol (0.0 to 12.4 gkg1 ). The total content of the other minor components was less than 10 gkg1 . All cultivars were suitable for hydro-alcoholic infusion production and, in spite of the minor oil yield, the leaves harvested in winter showed an essential oil composition quite similar to those from spring. KEYWORDS aromatic plants, cultivation, myrtle liqueur, gas chromatography, hydro-alcoholic infusions

Received September 21, 2010. The authors are grateful to the Ministero delle Politiche Agricole e Forestali (special grant IPPO) and to the Regione Autonoma della Sardegna for the nancial support of this research. Address correspondence to M. Mulas, Department of Economics and Tree Systems, University of Sassari, Via De Nicola, 9 07100 Sassari, Italy. E-mail: mmulas@uniss.it 21

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M. Mulas and R. A. M. Melis

INTRODUCTION
Myrtus communis (Myrtaceae) is an autochthonous species of the Mediterranean maquis. In recent years, researchers have focused on this plant species owing to increasing interest in the cultural and economic aspects of aromatic and medicinal plants (7). Myrtle leaves produce an essential oil that belongs chemically to two basic types: the cineoliferum type, rich in terpens (-pinene, limonene) and terpenoid oxides (1,8-cineole); and the myrtenilacetatiferum type, rich in terpenic esters (terpenil acetate, linalyl- acetate, bornyl acetate) and terpenoid oxides (1,8-cineole) (4). The yield and quality of the essential oil seem to depend on the geographical origin and season of collection, but the genotype of the species may also plays a role in its chemical variability (10). At present, the biomass of spontaneous myrtle shrubs is the only source of raw material for oil extraction and other industrial processes, such as the liqueur production from leaf hydro-alcoholic infusion (9). The sustainable use of natural resources and the conservation of myrtle habitat in the Mediterranean maquis is a problem. Steps must be taken to avoid degradation, reduction of genetic variability and, in the worst case, disappearance of the species (8). The increasing demand for raw material from industries that use myrtle leaves to produce a typical liqueur (White myrtle) increases the risk of impoverishing the natural maquis (11). The industrial demand for plant material raises the problems associated with the standardization of the nal product. Industry requires a known and denite quality of myrtle biomass, and this is not achieved with the picking of spontaneous vegetation. For all these reasons, a number of accessions from various locations in Sardinia (Italy) were collected and their characteristics studied. To establish their morphological types (68) and their rooting ability (5), they were grown in uniform eld conditions to eliminate the inuence of the environment, and their fruit and leaf yield were examined for their quality to evaluate their suitability for cultivation (11). After this domestication program, the present work reports the results of the qualitative and quantitative analyses of the essential oils of nine cultivars of Myrtus communis, as inuenced by year and season of sampling, and the correlations between these and the aroma of the corresponding infusions. The main objective of the research was the evaluation of the suitability of the leaf biomass produced by the new crop as raw material for the liqueur industry.

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MATERIAL AND METHODS Plant Material

Myrtle shrubs were grown in an experimental eld in Alghero (northwestern Sardinia, Italy) with planting distance of 3 m between rows and 1 m between plants along the row. Leaves from shrubs at the same state of maturity were picked from nine cultivars. Leaves were harvested from a random sample of 15 plants of each cultivar in Spring (May 2007 and 2008) and in winter (December 2007). The leaves of the young shoots were cut off, put in plastic bags, and frozen at 18 C until the time of oil extraction or infusion of the leaves. A subsample of the fresh material (10 g) was dried at 105 C for 24 h to check the water content.
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Essential Oil
The essential oils were extracted by hydro-distillation for 1 h from 200 g of frozen leaves in an all-glass Clevenger type apparatus using demineralized water (1 L). Three replicate extractions were performed for each cultivar. The oils were then recovered without using any solvent. The oils were stored in dark glass vials in a freezer at 20 C until GCMS and GC analysis.

Chemical Compound Identication

Identication of unknown components was performed in collaboration with the Interdepartmental Centre High Equipments of the University of Modena and Reggio Emilia (Italy) and by comparing their relative retention times and their mass spectra with those of data in GP and NIST libraries. The GC MS apparatus was a VARIAN SSQ 110 (model Advantage) equipped with a ame ionization detector and a fused silica capillary column (quartz crosslinked with 5% of phenyl-methyl-silicone), 25 m 0.30 mm i.d. 1 m lm thickness. The oven temperature program was set to rise from 60 C to 180 C at a rate of 4 C min1 . Injector and detector temperatures were 250 C. Hydrogen was used as the carrier gas (ow rate, 1 mL min1 ), with an air ow rate of 500 mL min1 . An oil sample of 0.3 L was injected in 90/10 split mode.

Analytical Gas Chromatography

The GC analysis was carried out using a Perkin Elmer GC apparatus (model Autosystem XL) equipped with a ame ionization detector and a fused silica capillary column, 60 m 0.25 mm i.d. 0.25 m lm thickness (DB-WAX, J &W Scientic). The oven temperature program was set to rise from 60 C to

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M. Mulas and R. A. M. Melis

200 C at a rate of 4 C min1 . Injector and detector temperatures were 250 C. Helium was used as the carrier gas (ow rate, 1 mL min1 ). An oil sample of 0.5 L was injected in the split mode 50/1. After identication of unknown components by GCMS, quantitative analysis was done by comparing their relative retention times and peak areas with those of the known authentic compounds previously injected in the GC column (all were Aldrich grade, except for -humulene, limonene, myrtenol, bornyl-acetate, camphene and 1,8-cineole, which were Fluka grade) and progressively added to the essential oil sample mixture (internal standard).

Infusions
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Ten g of frozen leaves was put in 80 mL of a hydro-alcoholic solution (70% ethyl alcohol) and stored in glass jars for 1 month at room temperature in the dark. At the end of the infusion time, the infusions were ltered. Ten mL of infusions was put in a glass vial, and 1 g of NaCl and 2 mL of a 1:1 ethylic ether:N-hexane solution was added to extract the avoring components. After agitation for 1 hour, the supernatant was separated with a pipette, and 2 L was injected in the gas chromatograph.

Statistical Analysis
All data presented are the means of three replications. Analysis of variance was performed with MSTAT-C software according to a factorial design to determine the inuence of the main variables (year, season, and cultivar). Duncans Multiple Range Test (p .05) was used to compare the means.

RESULTS AND DISCUSSION

The dry weight and the essential oil yield depended on the genotype (cultivar), on the phenological stage of the plant (season of collecting), and on the year of harvest (Figures 1 and 2). The leaf dry weight ranged from 332.6 gkg1 in cultivar Giuseppina to 411.8 gkg1 in cultivar Daniela in spring 2007; 397.2 gkg1 in Marta to 443.9 gkg1 in Daniela in winter 2007; and 307.2 gkg1 in Giuseppina to 386.7 gkg1 in Marta in spring 2008 (see Figure 1). The oil yield was calculated on the basis of the leaf dry weight. It ranged from 1.6 gkg1 in Angela to 10.7 gkg1 in Carla in spring 2007; 0.6 gkg1 in Marta to 4.2 gkg1 in Carla in spring 2008; and 0.8 gkg1 in Angela to 5.4 gkg1 in Barbara in winter 2007 (see Figure 2).

Essential Oil of Myrtle

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FIGURE 1 Mean dry leaf yield of nine Myrtle (Myrtus communis) cultivars. Mean separation by Duncans Multiple Range Test (p .05). Values followed by the same letters are not different.

It was also found that the myrtle oil composition depended on the phenological stage of plant and on the season of harvest, but the main source of variability seemed to be the cultivar. The main components detected in the essential oils were -pinene (ranging from 482.1 gkg1 in Daniela to 816.8 gkg1 in Tonina in spring 2007, from 406.5 gkg1 in Grazia to 789.8 gkg1 in Tonina in winter 2007, and from 366.8 gkg1 in Daniela to 714.7 gkg1 in Tonina in

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M. Mulas and R. A. M. Melis

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FIGURE 2 Essential oil yield extracted from leaves of nine Myrtle (Myrtus communis) cultivars. Mean separation by Duncans Multiple Range Test (p .05). Values followed by the same letters are not different.

spring 2008); limonene (ranging from 42.8 gkg1 in Tonina to 384.8 gkg1 in Daniela in spring 2007, from 52.9 gkg1 in Tonina to 397.3 gkg1 in Daniela in winter 2007, and from 56.1 gkg1 in Tonina to 468.8 gkg1 in Daniela in spring 2008);, 1,8-cineole (from 34.1 gkg1 in Angela to 221.8 gkg1 in Carla in spring 2007, from 58.1 gkg1 in Marta to 230.6 gkg1 in Barbara in winter 2007, ansd from 35.3 gkg1 in Daniela to

Essential Oil of Myrtle

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208.5 gkg1 in Carla in spring 2008); and linalool but with more than 10 gkg1 only in the cultivars Daniela (30.1 gkg1 in spring 2007, 30.4 gkg1 in winter 2007, 39.0 gkg1 in spring 2008), Carla (14.9 gkg1 in spring 2007, 51.9 gkg1 in winter 2008, 15.2 gkg1 in spring 2007) and, nally, Barbara (110.3 gkg1 in spring 2007, 56.6 gkg1 in spring 2008), geranylacetate (ranging from 12.4 gkg1 in Marta to 51.1 gkg1 in Nadia in spring 2007, 21.5 gkg1 in Marta to 99.2 gkg1 in Angela in winter 2007; 15.1 gkg1 in Daniela to 88.9 gkg1 in Marta in spring 2008) (Tables 14).The other components were terpinolene-isomer 1, present with more than 10 gkg1 in the cultivars Marta, Angela, and Giuseppina (Angela was the cultivar richest in terpinolene-1 with 41.0 gkg1 in spring 2007, 39.2 gkg1 in winter 2007, 38.3 gkg1 in spring 2008, followed by Marta:

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TABLE 1 Concentrations (gkg1 ) of -Pinene, Camphene, -Pinene, Isobutilbutyrate, and Limonene in the Leaf Essential Oil of Nine Myrtle (Myrtus communis) Cultivars Cultivar Spring 2007 Nadia Marta Barbara Daniela Angela Tonina Giuseppina Grazia Carla Winter 2007 Nadia Marta Barbara Daniela Angela Tonina Giuseppina Grazia Carla Spring 2008 Nadia Marta Barbara Daniela Angela Tonina Giuseppina Grazia Carla
Values

-pinene 634.4 751.0 602.5 482.1 718.4 816.8 796.3 503.6 508.2 612.9 707.9 501.7 420.3 515.0 789.8 719.0 406.5 597.5 557.3 394.0 573.0 366.8 537.8 714.7 685.8 394.7 441.6 de bc efg jkl c a ab ijk hij ef c ijk lmn hij ab c mn efg f-i mn e-h n g-j c cd mn klm

Camphene 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.4 0.4 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 a a a a a a a a a a a a a a a a a a a a a a a a a a a

-pinene 5.6 5.4 3.8 3.0 5.8 7.7 8.1 4.7 5.7 5.8 4.9 7.2 3.7 6.8 8.2 8.8 5.9 5.8 5.1 4.6 4.3 2.6 7.0 8.8 6.6 5.9 6.1 d-g d-h hij ij d-g abc ab gh d-g d-g fgh a-d hij b-e ab a d-g d-g e-h ghi ghi j bcd a b-f d-g c-g

Isobutylisobutyrate 4.3 2.4 2.6 4.8 4.5 2.8 3.3 6.6 8.6 1.8 0.0 5.2 3.7 2.6 1.1 1.1 1.8 4.1 5.8 0.0 2.7 7.2 4.0 4.0 0.0 4.6 13.3 d-h g-j ghi c-g d-g f-i e-i bcd b hij j c-f e-h ghi ij ij hij e-h cde j ghi bc e-h e-h j d-g a

Limonene 114.3 130.5 111.5 384.8 81.1 42.8 47.5 345.1 80.8 127.5 147.3 100.6 397.3 106.8 52.9 82.8 389.6 136.5 136.0 221.1 149.6 468.8 96.9 56.1 103.0 401.8 89.2 f-i efg ghi B K L L C K e-h E ijk B g-j L jk B ef ef D E A ijk L h-k B ijk

followed by same letters in each column are not different (Duncans Multiple Range Test, p .05).

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TABLE 2 Concentrations (gkg1 ) of 1,8-Cineole, p-Cymene, Terpinolene I, Terpinolene II, and Linalool in the Leaf Essential Oil of Nine Myrtle (Myrtus communis) Cultivars Cultivar Spring 2007 Nadia Marta Barbara Daniela Angela Tonina Giuseppina Grazia Carla Winter 2007 Nadia Marta Barbara Daniela Angela Tonina Giuseppina Grazia Carla Spring 2008 Nadia Marta Barbara Daniela Angela Tonina Giuseppina Grazia Carla
Values

1,8-cineole 127.5 48.8 99.0 39.8 34.1 49.9 55.9 62.3 221.8 124.0 58.1 230.6 80.0 78.1 59.4 78.9 107.6 126.4 130.6 95.1 110.7 35.3 88.7 86.4 61.3 104.5 208.5 bc ij c-f J j ij hij g-j a bcd g-j a e-i f-i g-j f-i b-f bc b def b-e j efg e-h g-j b-f a

p-cymene 0.0 2.0 2.6 2.0 6.7 1.9 2.0 4.1 2.1 3.5 3.7 10.8 7.7 13.0 4.3 5.4 8.1 9.6 1.1 2.2 5.3 2.5 3.6 1.3 3.8 3.7 4.0 m jkl g-l jkl cd kl jkl efg i-l f-k e-j b c a efg de c b lm h-l def g-l f-k lm e-i e-j e-h

Terpinolene I 2.0 14.7 2.2 0.0 41.0 8.7 9.5 7.6 0.0 1.2 19.1 0.9 0.0 39.2 14.5 15.1 14.1 1.5 9.0 42.6 4.3 1.2 38.3 10.1 26.3 11.9 1.8 i cde i i a gh fg gh i i c i i a cde cd def i g a hi i a efg b d-g i

Terpinolene II 1.1 2.8 6.8 8.8 7.4 0.7 0.6 4.4 9.9 0.7 0.7 5.2 3.0 0.9 0.0 0.0 2.1 2.4 3.4 10.8 13.0 13.8 11.4 4.2 6.1 6.4 14.9 klm i-l efg de efg lm lm g-j cd lm lm f-i i-l lm m m j-m j-m ijk bcd ab a bc hij fgh fgh a

Linalool 6.8 1.2 110.3 30.1 1.3 2.1 2.0 1.8 14.9 3.9 1.8 8.5 30.4 3.3 0.9 0.8 1.2 51.9 8.7 3.5 56.6 39.0 2.4 2.8 1.4 1.2 15.2 def f a c f ef ef f de def f def c def f f f b def def b c def def f f d

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followed by same letters in each column are not different (Duncans Multiple Range Test, p .05).

14.7 gkg1 , 19.1 gkg1 , and 42.6 gkg1 , respectively), -terpineol (with the largest content in Carla in spring 2007, 20.6 gkg1 ; 44.4 gkg1 in Angela in winter 2007; and 35.3 gkg1 in Angela in spring 2008), linalyl-acetate (never found in Daniela and Grazia; the maximum values were 24.7 gkg1 in Carla in spring 2007, 36.3 gkg1 in Angela in winter 2007, and 38.9 gkg1 in Carla in spring 2008). The content of other components was less than 10 gkg1 , with the exception of geraniol in spring 2008 (12.4 gkg1 in Marta), p-cymene in Angela in winter 2007 (13.0 gkg1 ), terpinolene-isomer-2 in spring 2008 in several cultivars, and -humulene (19.6 gkg1 in Marta in spring 2008). Camphene was present in traces (0.4 gkg1 ) in only the winter oils of two cultivars (Tonina and Giuseppina). Among all the compounds detected, -pinene was the prevalent component (>50%) in almost all the cultivars and in particular in Tonina,

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TABLE 3 Content (gkg1 ) of Linalyl-Acetate, Bornil-Acetate, -Humulene, Myrtenil-Acetate, and -Terpineol in the Leaf Essential Oil of Nine Myrtle (Myrtus communis) Cultivars Cultivar Spring 2007 Nadia Marta Barbara Daniela Angela Tonina Giuseppina Grazia Carla Winter 2007 Nadia Marta Barbara Daniela Angela Tonina Giuseppina Grazia Carla Spring 2008 Nadia Marta Barbara Daniela Angela Tonina Giuseppina Grazia Carla
Values

Linalylacetate 19.2 0.0 0.0 0.0 19.6 5.6 6.0 0.0 24.7 17.8 0.0 18.3 0.0 36.3 4.1 4.7 0.0 0.0 33.2 4.7 0.0 0.0 30.8 9.3 0.0 0.0 38.9 de g g g de fg fg g cd e g de g ab fg fg g g ab fg g g bc f g g a

Bornyl-acetate 2.0 0.0 0.0 1.0 0.0 0.0 0.0 0.0 2.2 2.1 0.0 1.7 0.7 0.0 0.0 0.0 0.0 0.0 4.2 0.0 0.0 1.6 0.0 0.0 0.8 0.0 3.9 b c c bc c c c c b b c bc bc c c c c c a c c bc c c bc c a

humulene 0.0 2.3 2.0 1.8 4.6 1.1 0.7 2.9 0.0 1.7 1.6 0.0 1.1 4.9 0.9 1.0 2.2 1.9 2.7 19.6 4.9 3.4 6.4 2.5 4.1 4.0 2.2 i e-h fgh f-i c ghi hi def i f-i f-i i ghi bc ghi ghi fgh fgh d-g a bc c-f b d-h cd cde fgh

Myrtenilacetate 0.0 0.0 1.4 0.0 0.0 0.0 0.0 1.4 0.0 0.7 0.5 0.0 2.2 0.8 0.9 1.6 2.2 2.1 0.0 1.5 1.6 0.9 0.0 0.5 1.3 2.2 0.0 b b ab b b b b ab b ab ab b a ab ab ab a a b ab ab ab b ab ab a b

-terpineol 11.4 1.8 4.3 2.2 9.6 6.8 8.2 1.8 20.6 13.4 1.8 17.8 2.5 44.4 7.4 9.9 2.0 2.9 10.8 4.9 2.7 1.6 35.3 9.9 2.0 2.2 21.8 def g efg fg d-g efg efg g c cde g cd fg a efg d-g fg fg d-g efg fg g b d-g fg fg c

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followed by same letters in each column are not different (Duncans Multiple Range Test, p .05).

Marta, and Giuseppina. The cultivars Daniela and Grazia had the lowest content of -pinene (<50%) but the greatest content of limonene (>35% against percentages <15% in others). Barbara, Carla, and Nadia were the richest in 1,8-cineole. Finally, the linalool content of Barbara was the highest (over 5% in the oils from spring leaves). The chromatograms of the infusion extracts showed a few peaks for the main components of the essential oils of leaves, namely -pinene, limonene, 1,8-cineole and linalool (Table 5). The content of -pinene ranged from 0.22 gkg1 in Daniela to 0.60 gkg1 in Carla in spring 2007, from 0.26 gkg1 in Angela to 0.52 gkg1 in Carla in winter 2007, and from 0.14 gkg1 in Daniela to 0.44 gkg1 in Barbara in spring 2008. The limonene ranged from 0.02 gkg1 in Marta to 0.14 gkg1 in Daniela and was not present in the infusions of Tonina

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TABLE 4 Content (gkg1 ) of Neryl-Acetate, Geranyl-Acetate, Myrtenol, Geraniol, and Metyl-Eugenol in the Leaf Essential Oil of Nine Myrtle (Myrtus communis) Cultivars Cultivar Spring 2007 Nadia Marta Barbara Daniela Angela Tonina Giuseppina Grazia Carla Winter 2007 Nadia Marta Barbara Daniela Angela Tonina Giuseppina Grazia Carla Spring 2008 Nadia Marta Barbara Daniela Angela Tonina Giuseppina Grazia Carla
Values

Nerylacetate 1.3 0.0 0.0 0.0 1.1 0.0 0.0 0.0 1.3 1.8 0.0 1.6 0.4 3.9 0.0 0.0 0.5 0.2 2.5 2.1 0.0 0.5 3.0 1.1 0.0 0.7 2.5 b-f f f f c-f f f f b-f b-f f b-f ef a f f ef f abc b-e f ef ab c-f f def a-d

Geranylacetate 51.1 12.4 18.6 15.1 27.4 31.2 35.8 12.9 55.0 62.0 21.5 59.2 25.1 99.2 28.4 38.9 24.5 31.6 63.0 88.9 23.3 15.1 82.1 52.1 23.4 25.1 76.1 def j hij ij g-j ghi fgh j de cd g-j d g-j a g-j efg g-j ghi cd ab g-j ij b def g-j g-j bc

Myrtenol 0.0 0.7 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.5 0.0 0.3 0.0 0.0 0.0 0.8 0.2 0.0 4.8 0.0 0.6 0.0 0.0 1.2 1.2 0.0 b b b b b b b b b b b b b b b b b b b a b b b b b b b

geraniol 0.0 2.1 3.8 0.0 0.0 0.0 0.0 0.0 1.5 0.0 1.3 1.3 1.2 0.6 0.5 0.4 1.2 2.1 0.0 12.4 2.6 1.4 0.2 1.9 1.2 1.5 1.7 f cd b f f f f f c-f f c-f c-f c-f def def def c-f cd f a bc c-f ef cde c-f c-f c-f

Metyleugenol 2.3 0.6 3.2 2.8 0.0 1.9 1.9 0.3 6.6 4.8 5.2 7.4 8.0 5.4 6.9 9.3 5.7 6.6 1.0 7.6 1.3 2.2 1.5 1.6 4.3 1.6 3.7 i-m lm g-k h-l m i-m i-m lm b-e d-h c-h abc ab c-g a-d a b-f b-e klm abc j-m i-m j-m j-m e-i j-m f-j

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followed by same letters in each column are not different (Duncans Multiple Range Test, p .05).

and Giuseppina in spring 2007; in spring 2008, it was detected only in the infusions of the cultivars Nadia and Grazia with contents of 0.08 gkg1 and 0.04 gkg1 , respectively. In winter 2007, limonene was present in four cultivars: Barbara and Carla (0.04 gkg1 ), Grazia (0.10 gkg1 ), and Daniela (0.16 gkg1 ). The 1,8-cineole ranged from 0.02 gkg1 in Angela to 0.2 gkg1 in Carla in spring 2007, whereas it was not present in the infusions of Daniela. In spring 2008, only Barbara and Carla showed a trace of 1,8-cineole (0.12 gkg1 ); in winter 2007, Marta and Angela were 1,8-cineolefree whereas Carla had the greatest content (0.16 gkg1 ). Linalool was present only in the infusion of Barbara in spring 2007 (0.08 gkg1 ).The presence of aroma compounds in the infusions seemed to be related to their presence in the essential oils of the leaves (Figure 3), although there was not a very close correlation between the quantities of

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TABLE 5 Components of the Flavor Extract of the Hydro-Alcoholic Leaf Infusions of Nine Myrtle (Myrtus communis) Cultivars (gkg1 ) Cultivar Spring 2007 Nadia Marta Barbara Daniela Angela Tonina Giuseppina Grazia Carla Winter 2007 Nadia Marta Barbara Daniela Angela Tonina Giuseppina Grazia Carla Spring 2008 Nadia Marta Barbara Daniela Angela Tonina Giuseppina Grazia Carla
Values

-pinene 0.36 0.38 0.38 0.22 0.30 0.32 0.50 0.32 0.60 0.34 0.32 0.46 0.36 0.26 0.44 0.46 0.34 0.52 0.24 0.24 0.44 0.14 0.16 0.34 0.22 0.20 0.36 abc abc abc abc abc abc abc abc a abc abc abc abc abc abc abc abc abc abc abc abc c bc abc bc bc abc

Limonene 0.06 0.02 0.04 0.14 0.04 0.00 0.00 0.12 0.08 0.00 0.00 0.04 0.16 0.00 0.00 0.00 0.10 0.04 0.08 0.00 0.00 0.00 0.00 0.00 0.00 0.04 0.00 a a a a a a a a a a a a a a a a a a a a a a a a a a a

1,8-cineole 0.08 0.06 0.06 0.00 0.02 0.06 0.04 0.06 0.20 0.08 0.00 0.08 0.04 0.00 0.02 0.06 0.08 0.16 0.00 0.00 0.12 0.00 0.00 0.00 0.00 0.00 0.12 a a a a a a a a a a a a a a a a a a a a a a a a a a a

Linalool 0.00 0.00 0.08 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 b b a b b b b b b b b b b b b b b b b b b b b b b b b

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followed by same letters in each column are not different (Duncans Multiple Range Test, p .05).

the single compound in the oils of each cultivar and the relative quantities in the corresponding infusion. It was found that the aroma of infusions depended on the year of collection and on the phenological stage of plants. Analysis of the essential oils conrms the existence of chemotypes that depend on the genotype of the plant. Each cultivar has a peculiar aromatic prole, but all the cultivars belong to the cineoliferum type (4). The myrtle oils extracted from leaves are rich in -pinene (up to 81.68%), limonene (up to 46.88%), 1,8-cineole (up to 22.18%), and linalool (up to 11.03%). The composition of the oils conrms the differences that exist between Sardinian oils and Spanish and Moroccan oils (13). Our oils are poor in myrtenyl-acetate and on average have greater -pinene content. The oil yields that have been reported for wild plants in Sardinia were about 0.2% to 1.6% on the basis of the dry weight (12). The essential oil

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M. Mulas and R. A. M. Melis

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FIGURE 3 Correlations between the content of avoring compounds in the leaf essential oils and in the hydro-alcoholic infusions obtained from leaves of nine Myrtle (Myrtus communis) cultivars.

Essential Oil of Myrtle

33

yields of the accessions are in the range registered for the wild plants, but it is well known that the total biomass produced had an inuence on the total oil yield. There is a need to establish whether horticultural practices can improve the biomass harvest and thus the absolute oil yield. Angela and Grazia are the cultivars in cultivation with the best biomass production (11), and their harvesting may protably replace the picking of spontaneous vegetation.

CONCLUSIONS
The composition of the essential oils inuences the aroma of the infusions; consequently, the main components of oils (-pinene, limonene, 1,8-cineole, linalool) were fundamental for oil quality. The main components of infusion aroma are in very low percentages because of the dilution of the components in comparison to the essential oils. The presence of -pinene, limonene, 1-8-cineole, and linalool is somewhat correlated to their relative abundance in the essential oils, but the differences among the cultivars do not seem to inuence in any signicant way the nal quality of the infusions and liqueurs. In other words, it seems that any of the cultivars may be used to produce liqueurs because the production technology attenuates the differences in the genotype of the raw materials. Moreover, our observation of differences between spring and winter leaves showed that, in spite of the lower essential oil yield from winter leaves, the oil composition is quite similar to that from spring leaves. This result may ensure a technological value to the high quantity of leaves usually harvested by mechanical harvesters together with myrtle fruits in winter. This by-product may be used for essential oil extraction or directly to produce leaf hydro-alcoholic infusions, which had results comparable to the ones obtained from the leaves harvested in spring.

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REFERENCES
1. Boelens, M. H., and R. Jimenez. 1992. The chemical composition of Spanish Myrtle oils. Part II. J. Essent. Oil Res. 4:349353. 2. Bradesi, P., F. Tomi, J. Casanova, J. Costa, and A. F. Bernardini. 1997. Chemical composition of Myrtle leaf essential oil from Corsica (France). J. Essent. Oil Res. 9:283288. 3. Lawrence, B. M. 1990. Progress in essential oils. Perfumer & Flavorist 15:6369. 4. Lawrence, B. M. 1993. Progress in essential oils. Perfumer & Flavorist 18:5255. 5. Mulas, M., and M. R. Cani. 1996. Variability of rooting ability of softwood cuttings in myrtle germplasm. In Proceedings of the International Symposium on Breeding Research on Medicinal and aromatic Plants. Quedlinburg, Germany, June 30July 4: 191194.

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6. Mulas, M., and M. R. Cani. 1999. Germplasm evaluation of spontaneous myrtle (Myrtus communis L.) for cultivar selection and crop development. JHSM 6(3):3149. 7. Mulas, M., and P. Deidda 1998. Domestication of woody plants from Mediterranean maquis to promote new crops for mountains land. Acta Horticult. 457:295301. 8. Mulas, M., M. R. Cani, and N. Brigaglia. 1998. Characters useful to cultivation in spontaneous populations of Myrtus communis L. Acta Horticult. 457:271278. 9. Mulas, M., M. R. Cani, N. Brigaglia, and P. Deidda. 1999. Study of myrtle (Myrtus communis L.) genetic resources to promote extensive crop as integration of spontaneous harvest. Acta Horticult. 502:8588. 10. Mulas, M., A. H. D. Francesconi, B. Perinu, and A. Fadda. 2002a. Barbara and Daniela: Two cultivars for myrtle berries production. Acta Horticult. 576:169 175. 11. Mulas, M., A. H. D. Francesconi, and B. Perinu. 2002b. Myrtle (Myrtus communis L.) as a new aromatic crop: Cultivar Selection. JHSMP 9(2/3):127131. 12. Pirisino, G., A. Mul, M. D. L. Moretti, and M. Satta. 1996. Studio della resa e della composizione chimica dellolio essenziale di Myrtus communis L. spontaneo di Cuglieri (Sardegna). Rivista italiana EPPOS 19:159169.