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Effect of Organ Type , Drying Methods, and Extraction Time on Yield and Essential Oil Components of Micromeria Barbata L

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Effect of Organ Type , Drying Methods, and Extraction Time on Yield and Essential Oil Components of Micromeria Barbata L Sept. 2014. Vol. 4, No.5 ISSN 2307-2083 International Journal of Research In Medical and Health Sciences © 2013-2014 IJRMHS & K.A.J. All rights reserved http://www.ijsk.org/ijrmhs.html EFFECT OF ORGAN TYPE , DRYING METHODS, AND EXTRACTION TIME ON YIELD AND ESSENTIAL OIL COMPONENTS OF MICROMERIA BARBATA L. BEFORE FLOWERING. SAER ALWAN1*, KHALED EL OMARI 2,3,4, ITAB SUKARIEH1, HYAM SOUFI1, CHARAFEDDINE JAMA3, SAMI ZREIKA1,4. 1Laboratory of Pharmacognosy, Faculty of Pharmacy, Jinan University, Tripoli, Lebanon. 2Health and environment microbiology Laboratory, Azm center for biotechnology research, EDST. Lebanese University, Tripoli, Lebanon. 3Unité Matériaux et Transformations UMET UMR CNRS 8207, Ecole Nationale Supérieure de Chimie de Lille, Université de Lille 1, France 4Faculty of Science – Lebanese University, Tripoli, Lebanon. *Corresponding author: [email protected] ABSTRACT Micromeria barbata of Lamiaceae family is a perennial medicinal and aromatic plant grown in the rocky areas of most east Mediterranean region. This study was conducted to determine the effects of organ type, drying methods, and extraction time on the yield and components of the essential oil extracted from this plant during non flowering period. Three organ types (leaves, stems and mix of both leaves and stems), two drying methods (3 days sun drying and 5 days shade drying), four extraction times (1, 2, 3 and 4 hours) were investigated. The essential oils from each organ type were extracted by hydro-distillation and the chemical components were analyzed by GC/MS. The yield and the components of the oil were significantly affected by these methods. The highest essential oil yield (2.65%) was obtained from leaves that are dried under the sun after 3 hours of extraction. The lowest essential oil yield (0.3%) was obtained from stems, sun drying after 1 hour of extraction. The percentage of pulegone that is toxic was strikingly and significantly lower than the percentage shown in other studies. The significance of the effect of drying on other essential oil components of each part of this plant is also discussed. Key words: Micromeria barbata, organ types, drying methods, extraction time, yield, chemical components. 1-INTRODUCTION of pulegone and limonene (Bakkour et al, 2012). Bakkour et al, also reported that M. Barbata have Micromeria is a genus of flowering plants that belongs antimicrobial and antioxidant activities. to Lamiaceae family. Micromeria is very aromatic, native to rocky areas of S.E. Europe and western Asia Aromatic plants are often dried before extraction to including Lebanon (Dudai et al, 2001; Marinkovic et al, reduce moisture content and allow long time storage in 2002,). Traditionally, the plant is used in alternative a stable condition. Proper drying of medicinal plants is medicine as herbal tea for the treatment of colds, fundamental to the achievement of a high quality abdominal pains, eye infections, high blood pressure, product. It has been showed that drying method had a and heart disorders due to the principal constituents of significant effect on oil content and composition of its essential oil (Al-Hamwi et al, 2011; Telci and aromatic plants (Okoh et al, 2008; Asekun et al, 2006; Ceylan, 2007; Dudai et al, 2001). The chemical Ahmadi et al, 2008). Jamshidi et al. (2004) reported that composition of Micromeria species was studied essential oil percentage and essential oil component of extensively by many researchers (Al-Hamwi et al, 2011; fennel were affected by duration of essential oil Formisano et al, 2007; Telci and Ceylan, 2007; Gulluce extraction. Few studies have been performed on et al, 2004; Dudai et al, 2001; Kirimer et al, 1993). The Micromeria barbata. The objective of the current study essential oil of M. fruticosa ssp Barbata was was to investigate the effect of drying methods, determined in our lab and others to be mainly composed extraction time, and organ type on essential oil chemical components of Micromeria barbata before flowering. 43 Sept. 2014. Vol. 4, No.5 ISSN 2307-2083 International Journal of Research In Medical and Health Sciences © 2013-2014 IJRMHS & K.A.J. All rights reserved http://www.ijsk.org/ijrmhs.html 2-MATERIALS AND METHODS 3-RESULTS AND DISCUSSION Plant Material Results showed that organ type, drying methods, and extraction time had a significant effect on the essential Micromeria barbata was collected from a field in Bekaa oil content of non flowering Micromeria barbata. The Sifrin, Dennieh in north Lebanon during the early highest essential oil percentage (2.65%) was obtained summer 2014 (height above sea level: 1350m, average from leaves that are dried under the sun after 3 hours of annual rainfall: 230mm, and mean annual temperature: extraction time (table 1). The lowest essential oil 18°C). The fresh non flowering plant materials were percentage (0.3%) was obtained from stems, sun drying carefully separated into leaves, stems, and mixed of after 1 hour of extraction. As shown in table 1, further stems and leaves. Two drying methods were increase in the duration of extraction time has no effect investigated: shade drying and sun drying. The first on the yield of essential oil of Micromeria barbata. group of samples (100gr each) was shade dried at room Jamshidi et al (2004) suggested that the maximum temperature for 5 days until reaching a constant weight. essential oil percentage was obtained after 2.5 hour The second group (50gr each) was dried under the sun from Foeniculum vulgare Mill. for 3 days until reaching a constant weight. Plant material was distributed as a thin layer to accelerate In contrast to our results, the reduction of volatile oil drying process. The maximum daily temperature during content of plant by the impact of high temperature which the drying methods were performed was between drying process has been reported by some scientists 30-35°C. (Okoh et al, (2008); Blanco et al, (2002); Sekeroglu et al, (2007)). However, our results showed that high Essential Oil Extraction. temperature exposure of plant material under sun drying in comparison with shade drying did not lead to the The dried plant materials were powdered and used for great loss of volatile oil which explains the increase of extraction by using a hydrodistillation technique during oil yield extracted from sun drying method in our study. 3 hours in an all-glass Clevenger type apparatus that This is supported by Sefidkon et al who reported that separates water from oil. The extracted oil was stored in high temperature drying method of Satureja hortesis L. a dark glass tube and kept under refrigeration between 2 plants had the highest essential oil yield. to 8°C until analysis. Extraction was carried out in triplicates. The relative amounts of each chemical component were determined by the GC-MS analysis (Table 2). The oil Gas Chromatography - Mass spectrometry from the fresh plant was richer in chemical (GC/MS). constituents than the dry plant . Our team reported the identification of a total of 17 chemical constituents from The analysis of the oils was carried out using a fresh plant (Bakkour et al 2012), while 11 constituents Shimadzu QP 2010 plus gas chromatography system were identified in this report. The result supports the connected to a mass selective detector DB-5ms 5% observation by some workers who reported that there phenyl-95% methylsiloxane (30m x 0.25mm internal could be a 50-fold reduction in chemical diameter, 0.25μm film thickness). Helium was the composition when plant materials are dried (Fatemeh carrier gas (constant flow rate, 0.7 ml/min). Oven et al, 2006; Raghavan et al, 1997). These 11 temperature was set at 46°C for 7 min, then components identified in the essential oil of Micromeria programmed until 182°C at a rate of 3°C/min with hold barbata are affected by different drying methods and time of 5 min, then to 280°C at 20°C/min and finally organ type comprising of 98.5 to 99.94 % of total oil maintained at 280°C for 5 min. Split injection was (Table 2). There were significant qualitative and conducted with a split ratio of 1:100 and volume quantitative differences between chemical components injected, 4 μl of the oil. The MS operating parameters of the essential oil according to the drying methods and were as follows: Ionization potential, 70 eV; interface organ types. The most significant common major temperature, 200°C and acquisition mass range; 50-800. components of essential oils were 2-Pinen-7-one (44.8- Relative percentage amounts of the essential oil 69.5%), Piperitone (7.9-24.6%), Pulegone (5.6-11.4%), components were evaluated from the total peak area Limonene (6.4-11.2 %), 1-Oct-3-ol (4.6-7.5%), Β- (TIC) by apparatus software. Identification of Mycrene (3.2-4.9%). Other minor common components components in the volatile oil was based on the were β-Pinene (0.6-2%) and 3-Octanol (0.9-1.5%), and comparison of their mass spectra and retention time finally, α-Pinene and Sabinene have less than 1% each. with literature data and by computer matching with NIST and WILEY libraries as well as by comparison of Some minor components such as β-Pinene had a notable the fragmentation pattern of the mass spectral data with amount exclusive of the organ type, the stem in both sun those reported in the literature. and shade drying method. The amount of β-Pinene in 44 Sept. 2014. Vol. 4, No.5 ISSN 2307-2083 International Journal of Research In Medical and Health Sciences © 2013-2014 IJRMHS & K.A.J. All rights reserved http://www.ijsk.org/ijrmhs.html the stem alone is at least 3 folds higher than the leaves 5- ACKNOWLEDGEMENTS and the mix of both leaves and stems.
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