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The Chemical Compositions and Antibacterial Activity of Eucalyptus Radiata Leaf Essential Oil

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The Chemical Compositions and Antibacterial Activity of Eucalyptus Radiata Leaf Essential Oil Archives of Pharmacology and Pharmaceutical Sciences Research Article: RD-PHA-10003 The Chemical Compositions and Antibacterial Activity of Eucalyptus Radiata Leaf Essential Oil Asare Ebenezer Aquisman*,1,2 Droepenu Eric Kwabena1,2 1Graduate School of Nuclear and Allied Sciences, University of Ghana, Ghana 2Faculty of Resource Science and Technology, University Malaysia Sarawak, Malaysia Abstract The aim of this present study was to assess the yield, chemical constituent and antibacterial activity of Eucalyptus radiata leaf essential oil. Hydrodistillation method was used to obtain the E. radiata essential oil. Higher yields were obtained in mature leaf of E. radiata plant as compared to young leaf. Gas chromatography coupled with mass spectrometry (GC/SM) was used to analyse the chemical composition of the essential oil. The major compounds identified were, 1,8-cineole (i.e. 65.9% in young leaf oil sample and 62.8% in mature leaf oil sample), α-terpineol (10.3% in young leaf oil sample and 11.1% in mature leaf oil sample) and limonene (9.3% in young leaf oil sample and 10.9% in mature leaf oil sample). In terms of commercial feasibility, the E. radiata essential oil leaf collected in Odoben, Central part of Ghana satisfies the requirement. The E. radiata essential oil leaf was found to have potential antimicrobial activity against all tested bacterial {Staphylococcus aureus, (Gram +ve), Escherichia coli (Gram–ve), Acinetobacter baumanni (Gram –ve), Klebsielia pneumonia (Gram +ve) and Exiguobacterium aquaticum (Gram +ve)}. Keywords: Eucalyptus radiata, Antibacterial activity, Major compounds, Hydrodistillation, GC-MS Introduction etc. are few examples. Consequently, additional study of Research on essential oil is wide and ascending. Nevertheless, prospects of the essential oils of the species of Myrtaceae among the countless essential oils studied, species affiliating family is necessitated.1,3,4 Especially; commercial, to the Myrtaceae family are most well received in terms of pharmaceutical purpose, and medicinal essential oils for characteristics. Essential oils from the family of Myrtaceae example Eucalyptus essential oil.2,5 The genus Eucalyptus are used in soaps, toothpastes and body creams according from the family of Myrtaceae has been planted for countless to Gillian DM & Luqman S, et al.1,2 The essential oil of of years as suggested by Luqman S & Ali B, et al.2,6 One tea tree (M. altrnifolia); nutmeg (M. fragrans); thyme (T. of the medicinal Eucalyptus essential oils understudied Vulgaris); clove (S. aromaticum); blue gum (E. globulus) is Eucalyptus rdiata. It is essential aromatic oil, used for different kinds of infectious conditions alone or in a mixture Citation: Asare Ebenezer Aquisman, Droepenu Eric Kwabena (2019) of other essential oils. It uses as anti-infective make it an The Chemical Compositions and Antibacterial Activity of Eucalyptus indicator of antimicrobial potential of the essential oil and Radiata Leaf Essential Oil, JArchives of Pharmacology and Pharmaceu- tical Sciences (APPS). RD-PHA-10003 its compounds. Nevertheless, the chemical composition and antimicrobial activities of Eucalyptus r. growing in Received Date: 28 June 2019; Accepted Date: 08 July 2019; Ghana is yet to be examined. A starting point of scientific Published Date: 11 July 2019 proof needs to be set out in order to better comprehend its *Corresponding author: Asare Ebenezer Aquisman, Graduate School therapeutic features, anti-infective ability and champion its of Nuclear and Allied Sciences, University of Ghana, Ghana, Faculty real use in medicinal practices. Therefore, this present study of Resource Science and Technology, University Malaysia Sarawak, was sketched to assess yield, chemical composition and Malaysia, Email [email protected] antimicrobial activity of Eucalyptus radiata leaf essential © 2019 - ReDelve. All Rights Reserved oil. Arch Pharmaco and Phar Sci(APPS)2019| volume 1 | issue 1 1 The Chemical Compositions and Antibacterial Activity of Eucalyptus Radiata Leaf Essential Oil Materials and methods auto – sampler at 24.79 psi and an inlet temperature of 250°C. Chromatographic separation was achieved by a BPX-5 capillary Plant collection column (29.5m×0.25mm i.d., 0.2μm film thickness) with a splitless The E. radiate fresh leaves were collected from a cultivated site injector and mass spectrometer detector. Oven temperature was -1 in Agona Odoben in the Central part of Ghana. Material samples programmed from 60°C to 2400C (5 min hold), at 6°C min , and -1 (Leaves) were identified and harvested under the assistance of local from 240°C to 300°C (15 min hold), at 6°C min rate. Helium was -1 farmer, Mr. Kojo Saah. To avoid loss of any volatile compounds used as the carrier gas (0.98ml min ). Spectra were achieved on all the plant material were processed within 32 hours. electron impact at 70eV, scanning from 35 to 550m/z. Each peak area of the gas chromatography components represented the % Preparation of Eucalptus radita leaf essential oil (percentage) composition of each individual component as a % (percentage) of all the peak zones (areas) obtained from electronic According to1,7 Eucalyptus is locally prepared in the form of poultices or infusions, which can be ingested as teas or used through integration computations using flame ionization detector (FID, inhalation. Infusions are usually for respiratory diseases whereas 250°C). 42 n-Alkanes were used as reference point in the poultices are for topical application. The medicinal application of measurement of relative retention indices (RRI). Eucalyptus essential oil is known for respiratory conditions. In Preparation of test samples this present study, the essential oil was obtained, as explained by Van Vuuren SF et al.8 using a Clevenger – type apparatus which Eucalyptus radiata essential oil extract was tested using disc was subject to hydrodistillation. Young and mature leaves were diffusion method on nutrient agar medium.9 This study was distilled independently throughout the extraction period. In short, performed by preparing serial diluted concentrations of 300mg/ a known amount of 500g of weighed fresh leaf plant material was mL, 150mg/mL, 75mg/mL, 35.5mg/mL, 17.75mg/mL and placed into 5000L round bottom flasks. About 850L of distilled 8.75mg/mL. was added into each round bottom flasks. Each round bottom flasks with a mixture of water and plant material was heated to Preparation of media (Bacteria broth) boiling point at a temperature of 100°C with the help of heating The bacteria of interest for the activity of the E. radiata essential mantle for 3 hours, 30 minutes. The heated mixture of plant and oils were obtained from the stock culture provided by Virology water produces water vapour and oil. The water helps in carrying Laboratory, UNIMAS (Universiti Malaysia Sarawak). Five out the volatile oils as well as prevention of overheating. The bacterial were cultured using Tryptone Soya broth (TSB) and water vapour and volatile oil mixture was condensed to liquid Tryptone Soya agar (TSA). The media (TSB and TSA) was form via the condensing tube. The water and essential oil mixture prepared based on the manufacturer’s instructions. A mass of run from the condenser and collected in the receiving tube of 2.50g of the dried broth was dissolved in 200.0mL distilled water, the Clevenger apparatus. Essential oil is immiscible in water, and then autoclaved at a temperature of 121°C for 18 minutes. following in a separation of the two liquids in the collecting tube. The bacterial was sub-cultured in a 10.0mL of broth, each in This permitted for easy segregation of the hydrosol and essential recommended glass vial bottle for 24.0 hours in an incubator with oil during collection. a shaker at a temperature of 37°C (normal body temperature). The essential oil yield was calculated using Equation 1, Turbidity (Optical Density/OD) of the bacterial broth after 24.0 hours was computed by UV Mini Spectrophotometer, model Percentage yield = Weight of essential oil/Weight of plant material * 100 1240 of Shimadzu brand at wavelength 575nm. The turbidity in Equation 1 the media was used to confirm the sterility of the media. It was compared to the nutrient broth standard until the desired Optical The assessed essential oil yield was recorded and stored in tightly Density of 0.6-0.9 was obtained. sealed amber bottles, away from light at 3°C for further analysis. Plate inoculation Chemical composition analysis The inoculation of the bacteria was performed by using Biohazard The essential oils were determined using Shimadzu Gas cabinent.9,10 1mL of the prepared broth was streaked over the entire Chromatography/Mass Spectrometer (GC/MS) QP 2010. A agar plate surface in four different directions using sterile cotton volume of 1µL was injected using a split ratio (200:1) with an bud. Before applying the test samples, the agar plate was then Arch Pharmaco and Phar Sci(APPS)2019| volume 1 | issue 1 The Chemical Compositions and Antibacterial Activity of Eucalyptus Radiata Leaf Essential Oil left for 5-10 minutes. A 10μl volume of test extract (Eucalyyptus Compound Young Mature r. essential oil) of concentrations 300mg/mL, 150mg/mL, 75mg/ α – Pinene 0.9±0.02 1.4±0.01 mL, 35.5mg/mL , 17.75mg/mL and 8.75mg/mL were each pupated onto the discs (6.0mm diameter) and gently pressed α – Thujene 2.4±0.01 1.9±0.02 onto the agar plate using sterile forceps for 12.0 minutes at 25°C β – Pinene 0.8±0.01 1.1±0.02 (room temperature). A disc pupated with methanol and 30.0μg of Sabinene 1.2±0.01 1.1±0.03 tetracycline as standard antibacterial agent were used as negative Mycrene 1.2±0.02 1.4±0.01 and positive controls respectively. Each of the test samples were tested in triplicate for the bacterium used. The plate samples were α – Terpinene 1.3±0.01 1.0±0.01 then incubated at a normal body temperature (37°C) for 24 hours Limonene* 9.3±0.02 10.9±0.07 before the inhibition zone of each sample disc investigated.
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