Journal of Plant Science and Phytopathology

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    Submitted: May 17, 2022 | Approved: June 13, 2022 | Published: June 14, 2022

    How to cite this article: Asfaw MD. Chemical composition of olive stems essential oil from Ethiopia. J Plant Sci Phytopathol. 2022; 6: 057-061.

    DOI: 10.29328/journal.jpsp.1001075

    Copyright License: © 2022 Asfaw MD. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

    Keywords: Olive; Essential oil; Dry distillation; Chemical composition; Olea Europaea

    Chemical composition of olive stems essential oil from Ethiopia

    Melese Damtew Asfaw*

    Department of Chemistry, College of Natural and Computational Sciences, Mekdela Amba University, P.O. Box 32, Ethiopia

    *Address for Correspondence: Melese Damtew Asfaw, Department of Chemistry, College of Natural and Computational Sciences, Mekdela Amba University, P.O. Box 32, Ethiopia, E-mail: [email protected]

    Abstract
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    In this article, the chemical compounds, antimicrobial and antioxidant activity of the volatile oil from leaves of Olea Europaea L. cultivar from Ethiopia has been studied. The essential oil was provided with a dry distillation apparatus and analyzed by GC-MS/FID. This analysis leads to the detection of 128 compounds representing 89.4% of the total oil. The major constituents were methyl ester hexadecanoic acid (4.10%), 2,4-dimethoxyphenolAa (4.05%), 2-methoxy-phenol (3.25%), 3,5-dimethoxy-4-hydroxytoluene (3.20%), 2-methoxy-5-methyl phenol (3.19%), 1,2,3-trimethoxy-5-methyl benzene (2.93%), 2-methoxy-4-vinyl phenol (2.70%), 2-hydroxy-3-methyl-2-cyclopenten-1-one (2.60%), trans-Isoeugenol (2.45%) and (E) -2,6-dimethoxy-4- (prop-1-en-1-yl) phenol (2.25%). The composition of essential oils was dominated by phenolic compounds.

    Introduction
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    Traditional medicine has been practiced in almost every culture, and it has spread worldwide and gained popularity [1]. In Ethiopia, knowledge of traditional medicine has been passed down from generation to generation, and about 80 percent of Ethiopians still rely on traditional medicine, especially for medicinal plants [2,3]. Essential oils are a complex mixture of variables commonly present in low concentrations and are essential components used for their flavor and aroma in the food, pharmaceutical, and perfume industries [4].

    Olea Europaea commonly called wild olive is found throughout the Mediterranean, Europe, Africa, Iran, Asia, and Ethiopia and is thought to have a farming history of several 1000 years [5]. It holds historical significance in the religious context and is quoted in Christian and Hebrew Bibles and the Koran [5,6]. The olive tree is rarely eaten as a natural fruit because of its bitter taste but is used as oil or table olive and its wild and cultivated forms are considered an important subject of plant research [5]. O. Europaea has been shown in traditional medicine. It has been known to lower blood sugar, cholesterol, and uric acid. It is also used to treat diabetes, high blood pressure, inflammation, diarrhea, respiratory and urinary tract infections, gastrointestinal diseases, asthma, hemorrhoids, rheumatism, laxative, mouthwash, and vasodilator. Many phenolic compounds, especially secoiridoids and iridoids [7] and their pharmacological functions have been a major focus of scientists for the past decade [8,9]. However, the essential oil of Olea Europaea grown in Ethiopia has never been investigated before. Therefore, the purpose of this study was to determine the chemical composition of the essential oils Olea Europaea growing in Ethiopia through GC / MS analysis and to make comparisons with the literature.

    Experimental

    Description of the study area: Woreilu is one of the 24 administrative districts in the South Wollo Zone of Amhara Region, Ethiopia. It is located at 36° 26' 0" – 39° 43' 0" E longitude and 10° 34' 0" – 10° 60' 0 " N latitude and 492km far from Addis Ababa, Ethiopia, 571 km from Bahir Dar, the capital city of Amhara Region, as well as 91km from Dessie, West of Zonal town. As of the 2007 Ethiopia census, Woreilu town had a population of 14,817 and a 71013-hectare total area. According to the Agricultural and Rural Development office of the Woreda, agro-ecologically, the woreda is classified as “Dega” which accounts for 82% while the remaining 18% is “Woina Dega”. Of the total number of 23 kebeles administrations, 20 are rural. In the Woreda, most Kebeles produce crops in the “Meher” season, six kebeles in both seasons, and only one kebele in the “Belg” seasons. The agro-climatic conditions of the Woreda ranged from moderate to high, with an average altitude of 2730m above sea level. Annual rainfall ranges from 766.2 to 1250 mm. which is usually inadequate (short in duration), poorly distributed, and highly variable in inter and intra seasons.

    Plant material

    The dried olive stems were randomly collected from the local market of Woreillu town, South Wollo district, Ethiopia, in May 2018. The authenticity of the plant material was done in the Department of Biology and Biodiversity Management, Wollo University. The extraction of the Essential oil was employed by a traditional method (dry distillation) which is not previously been published Figure 1.


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    Figure 1: Dry pieces of Olea europaea stem (the author).

    Isolation and characterization

    The dried stems of Olea Europaera were cut (cut) into small pieces (≈20 cm long), weighed, and washed under tap water to remove any foreign material and dried on laboratory benches in a well-ventilated room before EO. About 2.0 kg of small pieces were loaded into a clay pot, after which, the pot was turned into a cooking pot (cooking pot) and well mixed (tightly sealed) with mud so that it would not emit any steam from it (outside). Finally, the packed jar was buried in a hole 50 to 50 inches [50 × 50 cm] in diameter and set fire to it. EO collection started after a temperature of about 30 minutes and lasted for 1 hour until the clay pot became red hot. The hot pot was cooled for 10 minutes as it was in the oven and the flexible EO collected due to evaporation of the stew pot was separated from the charcoal by burning and stored in 250 ml solid glass containers. Finally, EO was refrigerated until it was needed for chemical analysis and bioassays testing.

    Stem EO analysis of Olea ertupaea was performed on a Shimadzu GC-2010 gas chromatograph with flame ionization detector (FID), inserted 25 m x 0.25 mm x 0.25 µm CBP5 capillary column, and using helium as a carrier gas. the oven temperature was set from 60 ºC (after 10 minutes) to 230 ºC at 3 ºC / min and the final temperature was 10 min. GC/MS analysis of stem EO of Olea ertupaea was performed with Agilent 5975N gas chromatograph-mass spectrometer with 30 m x 0.25 mm x 0.25 µm film thickness capillary column of HP5MS, using helium as a carrier gas. The oven temperature system was similar to that used in gas chromatography (GC) analysis.

    The chemical properties of Essential Oils have been identified by comparing their MS with the reference spectra at the National Institute of Standards and Technology (NIST) mass spectrometry data center and by comparing their storage indicators with Kovats' indications in the literature. Quantitative data were obtained electronically at a percentage of the area and peaks combined without the use of a corrective factor [10].

    Results and discussion
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    It was noted that the local oil yield of Olea europaea was 315.5 mL obtained from 6kg of plant material in three abortion groups yielding a yield of 5.19 ± 0.05 (% v/w) (Table 1).

    Table 1: Percentage yield (% v/w) of Olea europaea EO.
    Batch No. Weight of plant material(kg) Volume of Essential oil (mL) Percentage yield(v/w)
    1 2.0 105 5.25
    2 2.0 103 5.15
    3 2.0 103.5 5.18
     Mean ± SD = 5.19 ± 0.05

    The standard deviation of the three-group batch yield (% v/w) was found to be 0.05 equivalents to 0.96% of the standard deviation relative (% RSD). % Of RSD was used as an indicator of the accuracy of the dry distillation immersion process. The RSD% of this study, less than 2%, indicated that the dry distillation procedure was more accurate with less damage [11]. In addition, the distillation method used in this study yielded better results compared to other abortion techniques in classifying higher molecular terpenes such as diterpenes and triterpenes contradicting Birhanu's [12] study, which argued that diterpenes and higher terpenes cannot be detected by steam distillation method as these molecules are very heavy to allow evaporation, so they are rarely found in dissolved essential oils.

    One hundred and twenty-eight compounds comprise 89.4% of the essential oils identified by GC and GC/MS. Its main compounds were methyl ester hexadecanoic acid, 2,4-dimethoxyphenol, 2-methoxy-phenol, 3,5-dimethoxy-4-hydroxytoluene, 2-methoxy-5-methyl phenol, 1,2,3-trimethoxy -5-methyl benzene, 2-methoxy-4-vinyl phenol, 2-hydroxy-3-methyl-2-cyclopenten-1-one, trans-Isoeugenol and (E)-2, 6-dimethoxy-4- (prop-1- en- 1-yl) phenol, respectively (Figure 2, Table 2). To the best of my knowledge, this is the first report on the production of essential oils from Ethiopian Olea europaea. Phenolic compounds (35.49%), non-terpenes (29.23%), terpenes (20.90%), and other compounds (6.37%) dominated fat formation.

    The essential oil of Olea europaea contains compounds of interesting biological properties. Some authors stated that phenolic compounds and their analogs have strong antibacterial, antifungal, antiviral, anti-mutagenic, anti-inflammatory, and antioxidant activities [13,14]. This could well explain the importance of the Olea europaea in the traditional Ethiopian pharmacopeia.


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    Figure 2: The representative ion chromatogram of the major compositions of the stem essential oil of Olea Europaea.

    Table 2: The ten major compounds of the EO of the stem of Olea europaea.

    No Name of compounds Chemical formula Retention time Peaks Area (%) LRI* Class
    1 2-hydroxy-3-methyl-2-cyclopen-1-one C6H8O2 8.62 a 2.60 8.6178 Ketone
    2 2-methoxy phenol C7H8O2 9.35 b 3.25 9.3519 Phenol
    3 2-methoxy-5-methylphenol C8H10O2 10.85 c 3.19 10.851 Phenol
    4 2-methoxy-4-vinylphenol C9H10O2 12.76 d 2.70 12.763 Phenol
    5 2,4-dimethoxyphenol C8H10O3 13.50 e 4.05 13.503 Phenol
    6 trans-isoeugenol C10H12O2 14.48 f 2.45 14.477 Phenol
    7 3,5-dimethoxy-4-hydroxytolune C9H12O3 14.65 g 3.20 9.3519 Phenol
    8 1,2,3­trimethoxy-5-methyl benzene C10H14O3 15.54 h 2.93 15.545 Benzene
    9 (e)-2,6-dimethoxy-4-(prop-1-en-1-yl) phenol C11H14O3 17.02 i 2.25 17.681 Phenol
    10 Methyl ester hexadecanoic acid C17H24O2 18.45 j 4.10 18.44 Fatty acid

    This investigation is different from those found in some oils from Algeria (from leaves) [15] (Palmetic acid, Z-nerolidol, Octacosane), Tunisia (from fruits and stem) [16], (3-ethyl pyridine, (E)-2-decanal, 2-ethylbenzaldehyde, and Nonanal, (E, E) -2,4-decenal, Benzyl alcohol respectively) and South Africa (from leaves) (Iweriebor, et al. 2012) (Nonanal, Phytol, 2-isopropyl-5-methyl-9­methylenebicyclo[4.4.0]dec-1-ene. this variation in compositions and yield of the EO could be due to factors such as plant age, plant part, development stage, growing place, harvesting period, method of extraction, and principally by chemo-type since they influence the plant biosynthetic pathways and consequently the relative proportion of the main characteristic compounds [17] Table 3.

    Table 3: Chemical components of the stem oil of Olea europaea.
    PK Name of compounds LRI Area (%)
    1 ethanedioic acid, bis(1-methyl propyl) ester 4.3603 0.3862
    2 Silver butanoate 4.9696 0.0485
    3 3-Piperidinol, 1,4-dimethyl-, trans- 5.0419 0.1902
    4 Pyrazole, 1,4-dimethyl- 5.2979 0.7987
    5 2-Furanmethanol 5.9539 0.4980
    6 1,6:2,3-Dianhydro-4-O-acetyl-.beta.-d-mannopyranose 6.1198 0.3169
    7 2,4-Pentanedione, 3-methyl- 6.234 0.6179
    8 D-Limonene 6.6053 0.2173
    9 1,3-Cyclopentanedione 7.0359 0.3313
    10 [1,3,4]thiadiazol, 2-amino-5-(2-piperidin-1-ylethyl)- 7.0809 0.6856
    11 2,5-Hexanedione 7.2579 0.0920
    12 2-Furancarboxaldehyde, 5-methyl- 7.4851 0.2218
    13 Piperidine-4-carbonitrile 7.5745 0.5887
    14 2-Cyclopenten-1-one, 3-methyl- 7.7484 0.1425
    15 tetrahydro[2,2']bifuranyl-5-one 7.8526 0.6810
    16 2(5H)-Furanone 7.9479 0.7259
    17 2(5H)-Furanone, 5-methyl- 8.169 0.1052
    18 2H-Pyran, 3,4-dihydro-2-methoxy- 8.2595 0.1147
    19 2-Cyclopenten-1-one, 2-hydroxy-3-methyl- 8.6178 2.5996
    20 2-Furanone, 2,5-dihydro-3,5-dimethyl 8.8347 0.5642
    21 Phenol 9.0642 0.4353
    22 Phenol, 2-methoxy- 9.3519 3.2458
    23 Methyl ethyl cyclopentene 9.5105 0.1684
    24 Cyclohexane, (1-methylethylidene)- 9.6202 0.0984
    25 Phenol, 2-methyl- 9.7876 0.4697
    26 Cyclohexene, 1-methyl-4-(1-methylethyl)-, (R)- 9.873 0.0718
    27 2-Cyclopenten-1-one, 3-ethyl-2-hydroxy- 9.9341 0.4840
    28 Maltol 10.016 0.7629
    29 Naphthalene 10.205 0.5577
    30 Phenol, 3-methyl- 10.326 0.9758
    31 Phenol, 2-methoxy-3-methyl- 10.381 0.3767
    32 Oxirane, 3-hydroxypropyl- 10.424 0.6297
    33 Glycoluril 10.715 0.7149
    34 2-Methoxy-5-methylphenol 10.851 3.1932
    35 2H-Azepin-2-one, hexahydro-1-methyl- 10.931 0.5768
    36 Phenol, 2,4-dimethyl- 10.972 0.8009
    37 3,4-Dimethoxytoluene 11.157 0.4976
    38 Phenol, 2,4,6-trimethyl- 11.262 0.1204
    39 ethanone, 1-cyclohexyl- 11.298 0.0839
    40 Phenol, 2-ethyl- 11.491 0.5437
    41 Phenol, 4-ethyl- 11.55 0.4447
    42 Benzene, 1-(2-butenyl)-2,3-dimethyl- 11.664 0.1810
    43 4-Hydroxy-2,4,5-trimethyl-2,5-cyclohexadien-1-one 11.741 0.7174
    44 2(3H)-Furanone, 5-acetyldihydro- 11.816 0.2873
    45 Phenol, 2,4-dimethyl- 11.896 0.1728
    46 2-Pyridinealdoxime 12.016 2.2458
    47 2,4,6-Cycloheptatrien-1-one, 2-amino- 12.163 0.5262
    48 Acetic acid,1-methyl-3-(1,3,3-trimethyl-bicyclo[4.1.0]hept-2-yl)- 12.282 0.5426
    49 Naphthalene,1,2,3,4,4a,5,6,8a-octahydro-4a,8-dimethyl-2-(1- 12.353 0.1334
    50 4-Hydroxy-3-methyl benzoic acid, methyl ester 12.402 0.2437
    51 1,4:3,6-Dianhydro-.alpha.-d-glucopyranose 12.488 0.7493
    52 Cyclopentane, 2-methyl-1-methylene-3-(1-methylethenyl)- 12.57 0.2661
    53 2,4-Dimethylanisole 12.638 0.3567
    54 2-Methoxy-4-vinylphenol 12.763 2.6978
    55 Pentadecane 12.9 0.5852
    56 4-ethylbenzoic acid, 2-(1-adamantyl)ethyl ester 12.994 0.2205
    57 ethyl Vanillin 13.142 1.7299
    58 Naphthalene, 2,6-dimethyl- 13.204 0.2171
    59 Spirohexane-5-carboxylic acid, 1,1,2,2-tetramethyl-, methyl ester 13.25 0.1113
    60 5-Hydroxymethylfurfural 13.304 0.4006
    61 Catechol 13.361 1.2459
    62 Naphthalene, 2,6-dimethyl- 13.419 0.3924
    63 2,4-Dimethoxyphenol 13.503 4.0507
    64 Benzene, 1,2,3-trimethoxy-5-methyl- 13.562 0.2827
    65 ethanone, 1-(2,5-dimethoxyphenyl)- 13.642 0.3037
    66 Aromandendrene 13.714 0.3760
    67 Naphthalene, 1,2,3,4-tetrahydro-2,2,5,7-tetramethyl- 13.761 0.1581
    68 1,2-Benzenediol, 4-methyl- 13.831 1.6478
    69 Phenol, 3,4-dimethoxy- 13.941 0.9395
    70 2(3H)-Furanone, 3-acetyldihydro-3-methyl- 14.062 0.3197
    71 1,4-Benzenediol, 2,5-dimethyl- 14.109 0.3584
    72 1,7-Octadien-3-one, 2-methyl-6-methylene- 14.181 0.2387
    73 1,2-Benzenediol, 3-methyl- 14.339 1.1046
    74 Citral 14.405 0.3183
    75 trans-Isoeugenol 14.477 2.4466
    76 Methyleugenol 14.553 0.1495
    77 3,5-Dimethoxy-4-hydroxytoluene 14.649 3.2041
    78 Benzaldehyde, 3-hydroxy-4-methoxy- 14.774 1.4052
    79 m-ethylaminophenol 14.927 0.1873
    80 ethanone, 1-(2,3,4-trihydroxyphenyl)- 14.98 0.5293
    81 Benzene, 1-methyl-4-(methylsulfonyl)- 15.065 0.1777
    82 1,3-Benzenediol, 4,5-dimethyl- 15.223 0.7862
    83 Naphthalene, 1,4,6-trimethyl- 15.27 0.5307
    84 3-Acetyl-2,5-dimethyl furan 15.414 0.4634
    85 Benzene, 1,2,3-trimethoxy-5-methyl- 15.545 2.9307
    86 Benzoic acid, 4-hydroxy-3-methoxy-, methyl ester 15.693 0.4183
    87 ethanone, 1-[4-(methylthio)phenyl]- 15.758 0.6582
    88 5-Sec-butylpyrogallol 15.975 0.3256
    89 Benzeneethanol, 4-hydroxy- 16.049 0.2884
    90 Cyclohexanone, 2,5-dimethyl-2-(1-methylethenyl)- 16.119 0.0865
    91 3-tert-Butyl-4-hydroxyanisole 16.217 0.6369
    92 2-Propanone, 1-(4-hydroxy-3-methoxyphenyl)- 16.36 1.0852
    93 5,7-Dimethyl-1,3-diazaadamantan-6-one Hydrazone 16.443 1.9776
    94 1,4-Benzenediol, 2,3,5-trimethyl- 16.556 0.5361
    95 1,6-Dimethyl-4-ethylnaphthalene (Norcadalene) 16.668 0.1184
    96 N',N'''-Bis(6-nitro-4H-pyran-2-ylmethylene)-2,5-pyridinedicarbohydrazide 16.716 0.2386
    97 Dithiocarbonic acid,O-ethyl ester, methylene-S(IV)-trifluoromethyl est 16.813 0.0660
    98 Phenol, 4-(3-hydroxy-1-propenyl)-2-methoxy- 16.872 0.1100
    99 (e)-2,6-Dimethoxy-4-(prop-1-en-1-yl)phenol 17.017 0.9346
    100 tyrosol, acetate 17.151 0.4435
    101 1-Acenaphthylenol, 1,2-dihydro-1-methyl- 17.204 0.2000
    102 1H-Cycloprop[e]azulen-4-ol,decahydro-1,1,4,7-tetramethyl-,[1aR 17.286 0.4625
    103 1,3-Oxathiolane, 2-(4-chlorophenyl)-2-methyl- 17.413 0.5472
    104 5-Methyl-5,8-dihydro-1,4-naphthoquinone 17.518 0.4139
    105 Ketone, methyl 2-methyl-1-cyclohexen-1-yl, semicarbazone 17.592 0.2294
    106 (e)-2,6-Dimethoxy-4-(prop-1-en-1-yl)phenol 17.681 2.2473
    107 Benzenepropanol, 4-hydroxy-3-methoxy- 17.745 0.2394
    108 1,5,9-Undecatriene, 2,6,10-trimethyl-, (Z)- 17.808 0.8683
    109 2,6,10-Dodecatrien-1-ol, 3,7,11-trimethyl- 17.851 0.5235
    110 beta.-D-Mannofuranoside, farnesyl- 17.895 0.2757
    111 Benzaldehyde, 4-hydroxy-3,5-dimethoxy- 18.019 0.6421
    112 1,3,6,10-Cyclotetradecatetraene, 3,7,11-trimethyl-14-(1-methylethyl)-, 18.076 0.3685
    113 1,3,6,10-Cyclotetradecatetraene, 3,7,11-trimethyl-14-(1-methylethyl)-, 18.188 0.6376
    114 tricyclo[4.3.0.0(7,9)]non-3-ene,2,2,5,5,8,8-hexamethyl-, 18.324 0.5180
    115 Hexadecanoic acid, methyl ester 18.44 4.1051
    116 5,6-Azulenedimethanol,1,2,3,3a,8,8a-hexahydro-2,2,8-trimethyl- 18.52 0.3135
    117 Naphthalene, 2,3-dimethoxy- 18.598 1.3904
    118 Methyl 4-hydroxy-3,5-dimethoxybenzoate 18.693 0.4963
    119 Benzaldehyde, 3,4,5-trimethoxy- 18.761 1.1728
    120 1H-Cycloprop[e]azulene, decahydro-1,1,7-trimethyl-4-methylene- 18.96 0.4079
    121 Hexadecanenitrile 19.082 0.5492
    122 4-Hydroxy-2-methoxycinnamaldehyde 19.144 0.1379
    123 Benzenepropanoic acid, 2,5-dimethoxy- 19.215 1.1188
    124 1,3,6,10-Cyclotetradecatetraene, 3,7,11-trimethyl-14-(1-methylethyl)-, 19.3 0.5383
    125 Oxirane,2,2-dimethyl-3-(3,7,12,16,20-pentamethyl-3,7,11,15,19-henei 19.395 0.3530
    126 7H-Furo[3,2-g][1]benzopyran-7-one, 4-hydroxy- 19.492 0.1530
    127 .beta.-Humulene 19.543 0.3816
    128 3-Amino-7-methyl-1,2,4-benzotriazine 1,4-dioxide 19.597 0.0695
      total   89.395
    Conclusion
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    References

    The major components of the essential oil of the examined Olea europaea dry stems are methyl ester hexadecanoic acid, 2,4-dimethoxyphenol, 2-methoxy-phenol, 3,5-dimethoxy-4-hydroxytoluene, 2-methoxy-5-methyl phenol, 1,2,3-trimethoxy -5-methyl benzene, 2-methoxy-4-vinyl phenol. The essential oil of Olea europaea dry stems is a potential source of natural antioxidants and antibacterial compounds which are used for the treatment of various diseases caused by free radicals and microbes.

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    Title Abstract Introduction Result and discussions Conclusion
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