Volatile Oil Composition of the Berries of Juniperus macrocarpa Sibth. and Sm., Gathered in Spain, The

Journal of Essential Oil Research: JEOR, Jan/Feb 2005 by Velasco-Negueruela, A, Pérez-Alonso, M J, Palá-Paúl, J, Íñigo, A, López, G

Abstract

The steam distilled oil obtained from the berries of Juniperus macrocarpa gathered in the fossil sand dunes of Huelva, southwest Spain, was analyzed by GC and GC/MS in combination with retention indices. The oil contained α-pinene (59.8-70.5%) and myrcene (14.8-15.2%) as major constituents and moderate amounts of γ-muurolene (2.5-5.5%). In addition to these components, minor amounts of β-pinene (2.5-3.4%), limonene (1.1-1.3%), terpinolene (0.9-1.4%), β-caryophyllene (0.4-0.7%), γ-cadinene (0.1-1.1%), δ-cadinene (0.4-1.0%) and α-cadinol (0.2-0.4%) and lower quantities of diterpenes were also identified.

Key Word Index

Juniperus macrocarpa, Cupressaceae, essential oil composition, α-pinene, myrcene.

Introduction

The genus Juniperus L. belongs to the Cupressaceae family, comprising about 60 species (1) found from the northern hemisphere to the mountains of tropical Africa and West Indies. According to Flora lberica (2), two sections can be recognized for the Iberian Peninsula:

section Sabina Spach is composed of Juniperus phoenicea L. ssp. phoenicea;]. phoenicea L. ssp. turbinata (Guss.) Nyman; J. thurifera L. and J. sabina L.

section Juniperus L., comprising Juniperus communie L. ssp. communia; J. communis L. ssp. hemisphaerica (K. Presl) Nyman; J. communis L. ssp. alpina (Suter) Celak. J. oxycedrus L. ssp. oxycedrus; J. oxycedrus L. ssp. hadia (H. Gay) Debeaux; J. oxycedrus L. ssp. macrocarpa (Sm.) Ball and J. namcularis Gand. (=J. oxycedrus L. ssp. transtagana Franco). Recently Adams (3) based on leaf essential oils and RAPDs (Random Amplified Polymorphic DNAs) established the following species status for the Juniperus oxycedrus complex: J. oxycedrus L. J. badia H. Gay; J, macrocarpa Sibth. et Sm. and J. navicularis Gand.

As far as we know there is only one previous report on the oil analysis of the berries of J, macrocarpa. An oil from this taxon (4) gathered in southern Greece (island of Elaphonissos) contained cc-pinene (63.0%) and myrcene (8.8%) as major constituents. In this work we have examined the berry oil volatiles of J. macrocarpa gathered in southwest Spain (Huelva Province, in Atlantic fossil sand dunes) by GC and GC/MS in combination with retention indices.

Experimental

Plant material: The sites of collection, voucher specimen numbers and yields of the berries of J. macrocarpa are listed below:

1. Ginés 6, female plant, 10755GL, 29/3/2001, Huelva (Spain), Punta Umbria, yield 1.5% based on dry weight of sample.

2. Ginés 4, female plant, 10756GL, 29/3/2001, Huelva, Mazagón, Matalascañas, yield 1.0% based on diy weight of sample.

Voucher specimens have been deposited at the Herbarium of the Real Jardín Botánico de Madrid, Spain. Plant material was identified by Ginés López González from the Real Jardín Botánico de Madrid, Spain.

Isolation of volatile constituents: Fresh berries of J. macrocarpa were ground and steam distilled for 8 h in an all glass Clevenger-type apparatus and the plant material was suspended in a chamber above the boiling water. The oils were dried over anhydrous sodium sulfate and stored at 47deg;C in the dark. The exhausted berries were oven dried 48 h at 1007deg;C for the determination of oil yields.

Analyses: Analytical GC was carried out on a Varian 3300 gas Chromatograph fitted with a Silicone DB-I capillary column (50 m × 0.25 mm, film thickness 0.25 µm); carrier gas N2, flow rate 1.5 mL/min, split mode, temperature programmed 60°-240°C at 3°C/min. Injector temperature 250°C, detector used FID, detectortemperature 300°C. Injection volume for all samples was 0.1 µL. Quantitative data were obtained by electronic integration of area percents without the use of correction factors.

GC/MS analyses were carried out on a Hewlett Packard 5890 gas Chromatograph fitted with a phase bonded poly (5% diphenyl 95% dimethylsiloxane) silicone PTE5 capillary column (30 m × 0.25 mm, film thickness 0.25 µm). Carrier gas He, flow rate 1.5 mL/min. Temperature program regimen was 70°C, 2 min and then programmed to 2507deg;C at 2°C/min. Injector temperature 2507deg;C. The Chromatograph was coupled to an HP 5971 A mass selective detector (70 eV).

Component identification: Most constituents were identified by comparing their retention indices with those of authentic standards. The latter were purchased, synthesized or identified in oils of known composition. The fragmentation patterns of mass spectra were compared with those stored in the spectrometer data base using the NBS54K.L and WILEY. L built-in libraries and with those reported in the literature (5-8).

Results and Discussion

The components of the oils, the percentage of each constituent and the retention indices are summarized in Table I. The components are arranged in order of GC elution from the non-polar column. The monoterpenes of the oils of the berries of J. macrocarpa were dominated by α-pinene (59.8-70.5%) and myrcene (14.8-15.2%).The total of morioterpene components was 85.0-94.8%. The most important sesquiterpene component was shown to be γ-muurolene (2.5-5.5%). Sesquiterpene constituents amounted to 4.8-13.7%. Interestingly, most diterpenes found in our oil, such as abietadiene, abietatriene, manoyl oxide and sandaracopimarinal were also detected in the berry oil of J. oxycedrus (9). The total of diterpenes was 0.4-1.3%. According to bibliography, the chemical pattern in the berry oils of J. oxycedrus was to produce pinane hydrocarbons as major components (9,10). In the berry oils of J. badia the main compounds were found to be pinane hydrocarbons, germacrene D and manoyl oxide (11), and in the berry oil of J. macrocarpa gathered in Greece (4) the major components were shown to be α-pinene and myrcene. We have also reported about the oil composition of J. navicularis (12,13) whose major components were α-pinene (27.9-36.1%), α-phellandrene (0.9-11.1%) and limonene β-phellandrene (24.3-40.7%)