Gil L, Tadesse W, Tolosana E & Lpez R, Eds. Eucalyptus Species Management, History, Status and Trends in Ethiopia.

Proceedings from the Congress held in Addis Ababa. September 15th-17th, 2010

Essential Oil Yield from Leaves of Eucalyptus grandis and Eucalpyus saligna and their Commercial Applications
Sisay Feleke Forestry Research Center, Forest Products Utilization Research Office, P.O.B 2322, Addis Ababa, Ethiopia. Tell: +251 114402180, Fax: +251 114423686, e-mail: sisayfeleke@yahoo.co.uk

Abstract
Over 50 Eucalyptus species were introduced to Ethiopia for various purposes especially for fuel wood and construction. Additionally they have industrial uses in the production of particle and fiberboards, which are the main and major raw material consuming part of these species. The Eucalyptus species are rich with essential oils, which are used in pharmaceutical, perfumery, and various industries. The use of Eucalyptus oil for pharmaceutical industry depends on the oil chemical composition and the species as well, oils rich with cineol greater than 80% and in perfumery industry the oil to b rich with citronellal. In Ethiopia, by hydrodistillation mechanism, the essential oil produced at pilot scale from Eucalyptus globulus and Eucalyptus citrodora are used in the detergent factories. The oil production from these species made from waste or felled big trees. The production of essential oils from biological material depends on the extraction techniques, plant species, and geographical location, age of materials (maturity), parts of the plant, season, and climatic factors. The constituents of the oils are mainly monoterpene and sesquiterpene hydrocarbons, oxygenated compounds derived from these hydrocarbons (alcohols, aldehydes, esters, ethers, ketones, phenols, and oxides). In the production of Eucalyptus oil the major raw material is leaf and therefore the biomass production of the leaf per hectare to be maximized. Keywords: essential oil, Eucalyptus, monoterpens

Introduction
The genus Eucalyptus consists of over 500 species of trees. Eucalyptus varies in form from low shrubs and multi-stemmed trees less than 10 m in height to large single-stemmed trees more than 60 m tall. The production of lignotubers is a characteristic of many eucalyptuses and other tree species and this generally makes them respond to coppicing. The ability to grow Eucalyptus under a coppice system of management makes it suitable for its widespread all over the world. Thus, Eucalyptus grows under a wide range of climatic and edaphic conditions and this is one reason for the wide distribution of Eucalyptus into different countries in the world. Non-timber forest products cover all the biological material (other than industrial round wood and derived products) that may be extracted from the forest and can be utilized by the household, marketed, or have cultural or religious significance. For

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Gil L, Tadesse W, Tolosana E & Lpez R, Eds. Eucalyptus Species Management, History, Status and Trends in Ethiopia. Proceedings from the Congress held in Addis Ababa. September 15th-17th, 2010

the sake of information gap regarding to other use of Eucalyptus species with in the country this paper is limited to the volatile oil of this species. Over 50 species of Eucalyptus tree species were introduced to Ethiopia for various purposes. E. globulus and E. camaldulensis are two of the most widely planted species were in the country (Davidson 1995). Together with genus Pinus, the two genera account for the larger part of exotic plantations of which the Eucalyptus plantations account over 58% (EFAP 1994). Among the introduced Eucalyptus species, 11 of them are well adapted to the Ethiopian climate and 6 of them: E. camaldulensis, E. globulus, E. citrodora, E. saligna, E. grandsi, and E. triticornis are widely distributed over the country (Davidson 1995, Friis 1995).Like pines, eucalypts are grown for timber, fuel wood, construction, and pole. Besides the timber value of Eucalyptus, it has different values from honeybee flower to chemical production i.e. essential oil. The leaves of many Eucalyptus species are well known for their oil production and known as eucalyptol oil (Sheila and Mude 1995). Eucalyptus oils are obtained by steam distillation from the leaves of Eucalyptus and have aromas characteristic of the particular species. In most cases, the distillation of essential oil from Eucalyptus species come either from branches trimmed from the stems of eucalypts that are felled for timber, pole or fuel wood production. Steam for the distillation purpose is either generated in separate boiler in the case of large volume distiller or inbuilt within the tank in the case of small distiller (Yagodin et al. 1999). Oils produced from young leaves may vary from old ones in their physicochemical characteristics (Yagodin et al. 1999). Even trees within the same provenance may produce oils that are quite different to each other. In conifer species of the temperate zone, with an increase of tree age the volatile oil content drops in Pinus sibirica at the age of 20 -30 years has 3-3.5%, of 30-50 years ca 2% and when between 50-100 years 1.7%. Eucalyptus species vary morphologically from site to site and the oil production is heritable character (Davis 1989). The synthesis of essential oil components start in the young and growing buds of tree (Yagodin et al 1999). Yagodin et al (1999) recommend storage of leafy material for the production of biologically active compounds; this includes the essential oils as well, in summer time to be processed within 24hr and in wintertime in temperate regions up to five days for conifer species. Yields of oil from leaf vary somewhat between species but on a commercial scale are of the order of 1 percent on a "fresh" basis. Moreover, the production of an essential oil influenced by abiotic factors likes sunlight, soil character, and sloppiness, rooting system (Yagodin et al. 1999). Of more relevance to the economics of production is the yield of oil per hectare and this is dependent on the biomass production as well as the oil yield from the leaf. There may also be marked differences in oil yield and quality within a species according to the provenance origin of the seed (Haq et al. 2007). Volatile oils are composed of different groups of compounds: acids, alcohols, aldehydes, ethers, and ketons with various structures grouped in to monoterpens, sesquiterpens, and diterpens. The Eucalyptus oils are classified according to their

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Gil L, Tadesse W, Tolosana E & Lpez R, Eds. Eucalyptus Species Management, History, Status and Trends in Ethiopia. Proceedings from the Congress held in Addis Ababa. September 15th-17th, 2010

composition and main end-use to medicinal, perfumery and industrial. Of these, the most important in terms of volume of oil production and trade is the medicinal type, characterized by high cineole content in the oil and the perfumery oils that produced from Eucalyptus citriodora are in the greatest volume. The perfumery oil differs from the medicinal oils in containing citronellal, as the major constituent. The oil is employed in whole form for fragrance purposes, usually in the lower soaps, perfumes, and disinfectants. Eucalyptus oil for medicinal use: This type of oil is rich with cineol (not less than 70% of the oil) and free from some unwanted compositions like phellanderene and valeric acid. Cineol rich eucalypt oil mainly used in the treatment of bronchial infections. Cineole is the compound that gives the Eucalyptus oil its characteristic odour, accompanied by small amounts of other components that vary from species to species. The Eucalyptus oil for perfumery is the lemon-scented oil of Eucalyptus citriodora (citronellal variant) and the somewhat rose scented geranyl acetate rich leaf. In Ethiopia, the Eucalyptus citriodora oil distilled at the former Essential Oil Research Center now renamed to Wendo Genet Agricultural Research Center. The objective of this paper is demonstrates the essential oil yield of the two Eucalyptus species with respect to the age of leaves.
Table 1. Oil yield of some Eucalyptus species Species Major component (%) Oil yield on fresh weight basis (%)

Medicinal oils E. camaldulensis cineole, 10-90 0.3-2.8 E. globulus cineole, 60-85 0.7-2.4 E. tereticornis 0.9-1.0 cineole, 45-45 Perfumery and flavoring oils E. citriodora citronellal, 65-80 0.5-2.0 Source: Erich (1989) and Ermias et al, (2000).

Methodology
Munesa Shashemene Forest enterprise (renamed as Arsi Forest enterprise) is located south of Addis Ababa at 240 km. The altitudinal gradients range from 1900 to 2700 m (http://www.munessa-forest.uni-halle.de/location/). The area has a bimodal nature rainfall, with unreliable minor rainy season from March to May and a major rainy season from July to September. The mean annual precipitation is 1,075 mm and the mean annual temperature is 15C (Anonymous 2010). The forest has an estimated area ca 22,000 ha with over 6000 ha plantation forest and natural forest of over 15000 ha. The Eucalyptus plantation the enterprise has estimated to 1640 ha of Eucalyptus globulus, E. grandis, and E. saligna (Anonymous 1990).

Site description

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Gil L, Tadesse W, Tolosana E & Lpez R, Eds. Eucalyptus Species Management, History, Status and Trends in Ethiopia. Proceedings from the Congress held in Addis Ababa. September 15th-17th, 2010

Sample collection and method of analysis Samples of Eucalyptus grandis and Eucalyptus saligna leaves were collected from Munesa Shashemene Forest enterprise (renamed as Arsi Forest Enterprise) ages of 1, 4, and 25 years Eucalyptus stands. Collected fresh leaves were stored in a deep freezer until the oil distillation. The essential oil of collected samples was distilled by Clevenger type apparatus for 3hr. the oil yields (w/w) are given in table 1. The oils were analyzed by Hewlett-Packard 6890 GC series equipped with FID and capillary columns. The column temperature was programmed at 50 210 C at a rate of 3 C/min. The injector and detector temperature were 220 C and 270 C, respectively. The major components of the oils were identified by comparison of the retention time with those authenticate standards.

Results and Discussion

The leaves of Eucalyptus species at the distillation time have moisture contents of 45-50%. The oil yield varies from 1.1 to 2.13% presented in the table 2. The obtained oil was light yellowish. Results obtained from the analysis of the two Eucalyptus species shows that the major components of these species was pinene, the obtained result was completely in agreement with Ermias et al. (2000) for the same species.
Table 2. Eucalyptus leaf oil yield of different ages.
Characteristics Moisture content Age 2 E. grandis E. saligna 59.54 58 Age 4 E. grandis E. saligna 58.29 59.52 Age 25 E. grandis E. saligna 53.82 54.27 2.13 1.4645 +6.72 1.1 1.4720 +5.02

Oil yield, % 1.54 1.51 2 1.1 * Refractive index n22 1.4645 1.4720 1.4645 1.4720 d * Rotation angel +6.72 +5.02 +6.72 +5.02 Note: * the refractive index and rotation angle were measured for combined oil

Results obtained from the study of the two Eucalyptus species namely E. grandis and E. saligna -pinene was identified as a major component 58.6% and 32.6% respectively. The essential oils of the two studied species were rich with pinene; they can be used as industrial raw material the chromatographic analysis shows that the volatile oil obtained from the E. grandis is poor in composition and rich in 1, 8-cineole in contrary to reports of Ermias et al (2000). In the leaves of E. grandis and E. saligna oil yield between the three ages there is no yield variation. The obtained result is in contrary to the conifer species and theoretical statements of Yagodin et al (1999). The production of essential oil depends on the planting density and the yield of the species. The oil yield varies from report to report that for E. citriodora; Ermias et al (1999) stated 1% whereas Getnet (2004) reported 4%. Both authors sample sit collection was the same but detail-sampling parameters were not clear and factors

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Gil L, Tadesse W, Tolosana E & Lpez R, Eds. Eucalyptus Species Management, History, Status and Trends in Ethiopia. Proceedings from the Congress held in Addis Ababa. September 15th-17th, 2010

for such a variation is not clear. The oil yield depends on the geographical location, climatic factors, age (maturity of plants), parts of the plant etc. The country is importing essential oils of various plant sources from different countries, which are estimated over 36million Birr for almost 22000 tonnes of oils only for three months from May to August 2007. The CIF costs of 1kg essential oils vary from 161- 446 Birr (Ethiopian Custom Authority 2010). Among the import of essential oils, the majority of it, over 95%, to be consumed in the nonalcoholic or flavoured food additives. Even though the classification of import goods were not by plant source some of the products can be produced with in the country. One of the possibilities is that the production of Eucalyptus oil from the existing tree stands during the clear felling for poles and posts. Currently there are a number of private companies engaged in the production of essential oils from different biomass; however, these producers have no their own plantation farm. These may create a problem in sustainability of the products. The price of Eucalyptus citriodora oil is 513Birr per kg and 613Birr for Cympogogen citrates.

Conclusion
The oil of E. grandis and E. saligna can be used as industrial solvent as its major component is pinene. Report of Ermias et al (2000) that Eucalyptus species like E. globulus, E. viminalis, E. triticornis, E. camaldulensis rich with cineole, and E. urophylla and E. citriodora with citronella. Therefore the oil of these species can be used in either medicinal or perfumery with the fact that by controlling unwanted composing compounds in the oil for targeted purpose. The huge resource of the Eucalyptus species in the country and its use is mainly restricted to the wood only the leaf remains in the fields. Therefore, in such cases the waste material can be used in the production of essential oils. If intended in the essential oil production there should be taken further some investigation including
leaf biomass yield and oil yield per hectare per year of selected species like E. globulus, E. triticornis, and E. citriodora; selecting and establishing plantations of high yielding species/provenances; Improving the efficiency of distillation process/units: the efficiency of distiller is improved through design either to reduce energy consumption or to oil yield; and determining the most appropriate time of harvesting age and harvesting time the present result showed us that there is no variation in oil yield between the studied ages but on the other hand Yagodin et al (1999) on his report on Russian conifer species the yield affected by age.

Acknowledgments

The author would like to thank the Munesa-Shashemene Forest industry enterprise (Arsi Forest enterprise) for allowing me to collect fresh sample leaves. I would like to thank also Prof. Ermias Dagne for GC-MS analysis. I also thank Ato Getachew Desalegn for his comments and the anonymous reviewers for their valuable comment and editorial works.

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Gil L, Tadesse W, Tolosana E & Lpez R, Eds. Eucalyptus Species Management, History, Status and Trends in Ethiopia. Proceedings from the Congress held in Addis Ababa. September 15th-17th, 2010

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