Int J Curr Pharm Res, Vol 12, Issue 4, 139-142Original Article


GAS CHROMATOGRAPHY–MASS SPECTROMETRY ANALYSIS OF BIOACTIVE COMPOUNDS IN ESSENTIAL OILS OF LEAF OF EUODIA SUAVEOLENS SCHEFF

BOY RAHARDJO SIDHARTA1*, PATRICIUS KIANTO ATMODJO1

1Faculty of Biotechnology, Universitas Atma Jaya Yogyakarta, Yogyakarta, Indonesia
Email: [email protected]

Received: 20 Mar 2020, Revised and Accepted: 22 May 2020


ABSTRACT

Objective: Essential oils extracted from the leaf of Euodia suaveolens have been reported. However, there is no published data on comprehensive report on the chemical constituents of the essential oils and the methods utilized to extract the essential oils from the plant. This research aimed to reveal and determine the chemical constituents of essential oils from the leaf of E. suaveolens.

Methods: Essential oils of leaf of E. suaveolens were extracted by steam distillation and were analyzed utilizing Gas Chromatography-Mass Spectrometry (GC-MS) methods.

Results: The GC-MS analysis revealed the presence of twenty-five different chemical constituents from the essential oils. The main chemical constituents of essential oils extracted from E. suaveolens’ leaves were as follows menthofuran (50.38 %), p-mentha-1,8-diene (14.34 %), limonen (10.99 %), evodone (5.55 %), α-curcumene (4.65 %), globulol (1.88 %), longipinenepoxide (1.66 %), and linalool (1.40 %). This present research found three compounds, namely p-mentha-1,8-diene, globulol, and longipinenepoxide that have never been reported by any researchers working with this plant.

Conclusion: The results showed that E. suaveolens contains essential oils that are potential to be explored further and utilized as medicinal products against some ailments.

Keywords: GC-MS analysis, Euodia suaveolens, Bioactive compounds, Essential oils


INTRODUCTION

Indonesia has huge plant biodiversity that has been utilized by their ancestor in their daily life activities (ethnopharmaceutica). Many plant species have become the source of medicinal herbs and one of it is zodia (local name; Euodia suaveolens Scheff. is its scientific name, which is synonym with Euodia hortensis Forst.). The plant is originally growth in the West Papua and its vicinities such as Papua New Guinea, Samoa, Tonga, and Niue [1, 2].

Commonly, the plant was introduced to other places in Indonesia as ornamental and garden decorations. In some areas such as Sepik, Papua New Guinea scrapping of leaf added with water was drank to relief cold; in Solomon island crushed leaves were used to treat boils, roots and leaves were mixed with Areca catechu nuts and lime as malaria treatment; in Pacific islands infusion of barks or leaves were utilized to relief fever, leaves were chewed to treat toothache and remedial bathing; leaf decoction was a favorite treatment to cure stomachache, fever, and menstrual illness; decoction of barks was more potent to relief stomach ulcers, reduce menstrual and childbirth pains; leaves and in florescenses were worn as a personal decoration for social occasions; the woods were used as house construction [1].

Euodia suaveolens has been very popular as mosquito repellent in Indonesia, specifically against Aedes aegypti, mosquito species that became the vector of dengue fever [3, 4]. Some research topics were utilizing a combination of the plant extract with other herb extracts for instance lemon grass (Cymbopogon citratus) [5] or rosemary (Rosmarinus officinalis) [6]. E. suaveolens has been tested to eradicate red flour beetle (Tribolium castaneum) pest [7] and has also been determined its toxicity utilizing Brine Shrimp Lethality Test (BSLT) [8]. Many research activities on E. suaveolens were done to find bioactive compounds and produce several antimosquito formulations, namely lotion, gel, electric mat, etc [3]. Some other researchers have been trying to prove its bioactive compounds as antibacterial l [9, 10].

Research on essential oils has been increasingly done by many researchers, since the chemical constituents in the essential oils showed important aspects to human life [11–13]. Some chemical constituents of the essential oils extracted from plants have been utilized not only as fragrances, flavors, and perfumes but also as antibacterial, antifungal, antioxidant, antitumor, anticancer, anti-inflammatory, etc [14-18]. However, published data on the chemical constituents of the essential oils from the leaf of E. suaveolens are dearth. Therefore, this research aimed to reveal the chemical constituents of the essential oils from the leaf of Euodia suaveolens and to determine their potencies as pharmaceutical resources.

MATERIALS AND METHODS

Preparation of materials

Leaves of E. suaveolens were collected from Sleman district, Yogyakarta province, Indonesia. The plant was authenticated and identified by plant taxonomist at Faculty of Biotechnology, Universitas Atma Jaya Yogyakarta and was deposited as herbarium at the Faculty of Biotechnology, Universitas Atma Jaya Yogyakarta. The leaves were selected based on good quality such as green in color, no insect bites, clean (no dirt), and fresh conditions.

Experimental procedures

Fresh E. suaveolens leaves sample (10 kg) were collected, washed thoroughly, decanted to reduce the water, and put into distillation apparatus and were steamed for four hours to extract its essential oils. Ten kg of the leaf sample was put into a biomass holding chamber of the distillation unit. The water steam was produced by heating the water in a vessel and the steam flows into a chamber where the leaf sample was put. The steam passed through the leaf samples and created vapor, which then condensed further and flowed into a receiver vessel. The essential oils layer was separated and kept in a clean bottle [11, 19, 20].

The essential oils were then analyzed utilizing GCMS-QP2010S Shimadzu (Shimadzu Ltd., Japan) equipment with the following conditions: column type Rtx 5, column length: 30 m, film: 0.25 μm, carrier gas: Helium, ionizer: EI 70 Ev, column oven temperature: 50.0 °C, injection temperature: 300.0 °C, injection mode: split, flow control mode: pressure, pressure: 13.0 kPa, total flow: 79.3 ml/min, column flow: 0.55 ml/min, linear velocity: 26.8 cm/sec, purge flow: 3.0 ml/min, split ratio: 139.0, ion source temperature: 250.00 °C, interface temperature: 300.00 °C, solvent cut time: 3.00 min, detector gain mode: Absolute, detector gain: 1.50 kV, threshold: 0. The chemical constituents of the essential oil were identified by comparing the results of the chromatogram and retention time reference from Wiley mass spectra library (Wiley229. LIB) [21-23].

RESULTS AND DISCUSSION

GC-MS chromatogram of the main chemical constituents of essential oils extracted from E. suaveolens’ leaves were as follows menthofuran (50.38 %), p-mentha-1,8-diene (14.34 %), limonen (10.99 %), evodone (5.55 %), α-curcumene (4.65 %), globulol (1.88 %), longipinenepoxide (1.66 %), and linalool (1.40 %). GC-MS analysis found 25 chemical constituents from the essential oils sample examined (table 1). Some of the chemical constituents identified were reported by earlier researchers to have bioactive applications such as antibacterial, antifungal, antitumor, anticancer, anti-inflammatory, etc. (table 2). These chemical constituents showed high potencies to be explored further as pharmaceutical products.

The chemical constituents revealed in table 1 were also found in the essential oils of Euodia sauveolens (and its synonym Euodia hortensis) in some researchs, but with some degree of differences. Both similarities and differences were found in term of chemical species and percentage area of the chemical constituents reported (table 3). Menthofuran has the highest percentage area and similar findings were reported by some researchers [9, 10, 24]. Other chemical constituents that were revealed by some researchers were limonen, evodone, α-curcumene, and linalool [1, 9, 10, 24, 25]. But this present research found three compounds namely p-mentha-1,8-diene, globulol, and longipinenepoxide that have never been reported by any researchers working with this plant. Steam distillation was suggested to be able to extract more chemical constituents from the plant as has been reported by some researchers [11, 20]. Further research on these three compounds will be interesting to be done, specifically to determine their bioactivities in health care applications.

The differences in the percentage area of the chromatogram reported by some researchers was due to the differences in the research method as well as the analysis method applied [13]. Some researchers noticed that the differences might probably be caused by the difference of plant origins (geography), environment conditions (ecology), growth conditions (physiology) [9], and plant species collected (genetics) [1, 2].

Table 1: Chemical compounds identified from the essential oils of E. suaveolens

No. Name of compounds RT Formula Molecular weight
1. 1,6-Octadiene, 7-methyl-3-methylene-(CAS) 2-Methyl-6-Methylene-2,7-Octadiene 13.111 C10H16 136
2. Cyclohexene, 1-methyl-4-(1-methylethenyl)-(CAS) 1-P-Mentha-1,8-Diene 14.598 C10H16 136
3. Benzofuran, 4,5,6,7-tetrahydro-3,6-dimethyl-(CAS) Menthofuran 19.454 C10H14O 150
4. 1-Cyclohexene-1-methanol, 4-(1-methylethenyl)-(CAS) Perillol 20.472 C10H16O 152
5. Evodone 24.951 C10H12O2 164
6. Bicyclo[3.2.1]octan-3-one 25.118 C8H12O 124
7. Tricyclo[4.4.0.0(2,7)]dec-3-ene, 1,3-dimethyl-8-(1-methylethyl)-, stereoisomer (CAS) Tricyclo[4.4.0.0(2,7)]dec-3-ene, 1,3-dimethyl-8-(1-methylethyl)-, st (CAS) Copaene 25.823 C15H24 204
8. Limonen-10-yl acetate 26.740 C12H18O2 194
9. Cyclohexanol, 2-methyl-5-(1-methylethenyl)-, acetate, (1. alpha.,2. beta.,5. alpha.)-(CAS) Dihydrocarveol acetate 26.958 C12H20O2 196
10. Bicyclo[7.2.0]undec-4-ene, 4,11,11-trimethyl-8-methylene-, [1R-(1R*,4E,9S*)]-(CAS) l-Caryophyllene 27.136 C15H24 204
11. Trans-Carvyl Acetate 27.358 C12H18O2 194
12. beta.-Caryophyllene 28.098 C15H24 204
13. 1,3,6,10-Dodecatetraene, 3,7,11-trimethyl-(CAS). alpha.-Farnesene 28.550 C15H24 204
14. Benzene, 1-(1,5-dimethyl-4-hexenyl)-4-methyl-(CAS) ar-Curcumene 28.668 C15H22 202
15. 3a,7-Methano-3aH-cyclopentacyclooctene, 1,4,5,6,7,8,9,9a-octahydro-1,1,7-trimethyl-, [3aR-(3a. alpha.,7. alpha.,9a. beta.)]-(CAS) Cloven 28.998 C15H24 204
16. Longipinenepoxide 29.303 C15H24O 220
17. 1-Cyclohexene-1-carboxaldehyde, 2,6,6-trimethyl-(CAS) 1-Formyl-2,6,6-trimethyl-1-cyclohexene 31.159 C10H16O 152
18. 1,4,8-Cycloundecatriene, 2,6,6,9-tetramethyl-, (E,E,E)-(CAS) 4,7,10-Cycloundecatriene, 1,1,4,8-Tetramethyl-, ALL-CIS 31.300 C15H24 204
19. 1,6-Octadien-3-ol, 3,7-dimethyl-(CAS) Linalool 31.491 C10H18O 154
20. Benzene, 1-(1,5-dimethyl-4-hexenyl)-4-methyl-(CAS) ar-Curcumene 31.592 C15H22 202
21. Aromadendrenepoxide-(II) 31.814 C15H24O 220
22. 7-Oxabicyclo 4.1.0 heptane, 1-methyl-4-(1-methylethenyl)- 32.173 C10H16O 152
23. Globulol 32.552 C15H26O 222
24. 2,6-Octadien-1-ol, 3,7-dimethyl-, (E)-(CAS) Guaniol 33.907 C10H18O 154
25. 3,5-Octadiene, 4,5-diethyl-3,6-dimethyl-(CAS) 47.681 C14H26 194

Note: RT= Retention time

Table 2: Bioactivity of chemical constituents identified from essential oils of E. suaveolens leaf

No. Compound name Bioactivity Reference
1. Menthofuran antioxidant [12]
2. p-mentha-1,8-diene insect repellent, flavoring ingredients [12]
3. Limonen antitumor, anticancer, antioxidant, anti-inflammatory, anti-stress [16]
4. Evodone insect (mosquito) repellent [9, 24]
5. α-Curcumene flavoring ingredients, perfumes and fragrances, insect repellent [18]
6. Globulol antimicrobial, antifungal, antibacterial [15]
7. Linalool cosmetics, perfumes and fragrances, biocides (disinfectants, pest control) [14]

Table 3: Comparison of chemical constituents of essential oils extracted from E. suaveolens leaf

No. Compound name Method, plant, country Researcher (s)
1. menthofuran, evodone (4-ketomenthofuran), limonene steam distillation, GLC analyzed, E. hortensis, Fiji [24]
2. prenilated acetophenon, monoterpene furano (evodone), caryophyllene, α-copaen, ar-curcumene method not mentioned, E. hortensis, Papua New Guinea [1]
3. evodone, menthofuran, limonene, curcumene, fonenol steam distillation, GC-MS analyzed, E. suaveolens, Indonesia [10]
4. limonene Ethanol maceration and hydrodistillation, E. suaveolens, Indonesia [25]
5. menthofuran, evodone, linalool, citronellol, α-(2) gurjunene hydrodistillation, GCMS analyzed, E. hortensis, Fiji [9]
6. linalool steam distillation and TLC analyzed, E. suaveolens, Indonesia [4]
7. menthofuran, p-mentha-1,8-diene, limonen, evodone, α-curcumene, globulol, longipinenepoxide, linalool steam distillation, GCMS analyzed, E. suaveolens, Indonesia Sidharta and Atmodjo (this present research)

CONCLUSION

This present research found twenty-five chemical constituents of essential oils identified by GC-MS from the leaf of E. suaveolens, which is higher in numbers of compounds compares to earlier research findings. The results also showed that E. suaveolens contains essential oils that are potential to be explored and utilized as medication to some ailments. These findings give strong support in term of scientific bases to the practices done by the ancestors (ethnopharmaceutical) to the plant. Therefore, continuous research on the essential oils of the plant will bring more results and more chances to utilize as pharmaceutical products that will give benefits to more peoples.

ACKNOWLEDGMENT

The authors thanked to Ms. Christiana Asmaranti Kirana Putri and Ms. Sri Wahyuni, our research assistants, for the laboratory works.

ABBREVIATIONS

cm= centimeter, Ev= electron volt, kg= kilogram, kPa= kilo Pascal, kV= kilovolt, m= meter, min= minute, ml= millilitre, sec= second, μm= micrometer

FUNDING

Research and Community Service Office Universitas Atma Jaya Yogyakarta partially funding the research.

AUTHORS CONTRIBUTIONS

Both authors have contributed equally.

CONFLICT OF INTERESTS

Authors declared no conflict of interests.

REFERENCES

  1. Djamal J. Euodia hortensis. JR forster, JG Forster. In: Lemmens RHMJ and Bunyapraphatsara N. (Editors). Plant Resources of South-East Asia, No. 12, Medicinal and Poisonous Plants 3, Backhuys Publishers, Leiden; 2003. p. 196-7.

  2. Hassler M. World plants: synonymic checklists of the vascular plants of the world. In: Roskov Y, Ower G, Orrell T, Nicolson D, Bailly N, Kirk PM, Bourgoin T, DeWalt RE, Decock W, Nieukerken E van, Zarucchi J, Penev L. (Editors). Species 2000 and ITIS Catalogue of Life, Annual Checklist, Naturalis, Leiden, the Netherlands; 2019.

  3. Simaremare EV, Sinaga DI, Agustini V. Sabun zodia sebagai Repellent terhadap nyamuk Aedes aegypti. Pharm J Indones 2017;3:11-6.

  4. Lestari FD, Simaremare ES. Uji potensi minyak atsiri daun zodia (Evodia suaveolens Scheff.) sebagai insektisida nyamuk Aedes aegypti L dengan metode elektrik. Pharmacy 2017;14:1-10.

  5. Mirawati P, Simaremare ES, Pratiwi RD. Uji efektivitas repellent sediaan lotion kombinasi minyak atsiri daun zodia (Evodia suaveolens Scheff.) dan minyak atsiri batang serai (Cymbopogon citratus) terhadap nyamuk Aedes aegypti L. Pharmacy 2018;15:1-15.

  6. Widawati M, Santi M. The effectiveness of fixative addition on zodia (Evodia suaveolens S.) and rosemary (Rosmarinus officinalis L.) gel against Aedes aegypti. Health Sci Indones 2013;4:103-6.

  7. Cameron RR, Arinafril, Mulawarman. Uji bioaktivitas ekstrak daun zodea (Evodia suaveolens Sheff) terhadap hama gudang Tribolium castaneum (Coleoptera: Tenebrionidae) herbst. E J Agroekoteknologi Tropika 2016;5:222-31.

  8. Lestari MS, Himawan T, Abadi AL, Retnowati R. Toxicity and phytochemistry test of methanol extract of several plants from papua using Brine Shrimp Lethality Test (BSLT). J Chem Pharm Res 2015;7:866-72.

  9. Chand R, Jokhan A, Gopalan R. Bioactivity of selected essential oils from medicinal plants found in Fiji against the spiralling whiteflies (Aleurodicus dispersus Russell). Adv Hort Sci 2017;30:165-74.

  10. Maryuni AE. Isolasi dan identifikasi senyawa antibakteri minyak atsiri daun zodia (Evodia sp). Sekolah Pascasarjana, Institut Pertanian Bogor, Bogor. Thesis; 2008.

  11. Bozovic M, Navarra A, Garzoli S, Pepi F, Ragno R. Esential oils extraction: a 24 h steam distillation systematic methodology. Nat Prod Res 2017;31:2387-96.

  12. Tisserand R, Young R. Essential oil profiles. http://dx.doi.org/10.1016/B978-0-443-06241-4.00013-8,2014.

  13. Lahlou M. Methods to study the phytochemistry and bioactivity of essential oils. Phytother Res 2004;18:435–48.

  14. Ung CY, White JML, White IR, Banerjee P, McFadden JP. Patch testing with the European baseline series fragrance markers. Br J Dermatol 2017;178:776-80.

  15. Tan ML, Zhou LG, Huang YF, Wang Y, Hao XJ, Wang JG. Antimicrobial activity of globulol isolated from the fruits of Eucalyptus globulus Labill. J Nat Prod Res 2008;22:569-75.

  16. Aggarwala BB, Shishodia S. Molecular targets of dietary agents for prevention and therapy of cancer. Biochem Pharm 2006;71:1397-421.

  17. Alviano WS, Mendonca Filho RR, Alviano DS, Bizzo HR, Souto Padron T, Rodrigues ML, et al. Antimicrobial activity of Croton cajucara benth linalool-rich essential oil on artificial biofilms and planktonic microorganisms. Oral Microbiol Immunol 2005;20:101–5.

  18. Antonious GF, Kochhar TS. Zingiberene and curcumene in wild tomato. J Environ Sci Health Part B 2003;38:489-500.

  19. Filly A, Fabiano Tixier AS, Louis C, Fernandez X, Chemat F. Water as a green solvent combined with different techniques for extraction of essential oil from lavender flowers. C R Chimie 2016;19:707-17.

  20. Ozek G, Ozek T, Iscan G, Baser KHC, Hamzaoglu E, Duran A. Comparison of hydrodistillation and microdistillation methods for the analysis of fruit volatiles of Prangos pabularia lindl., and evaluation of its antimicrobial activity. South Afr J Bot 2007;73:563–9.

  21. Amelia B, Saepudin E, Cahyana AH, Rahayu DU, Sulistyoningrum AS, Haib J. GC-MS analysis of clove (Syzygium aromaticum) bud essential oil from Java and Manado. AIP Conf Proc 2017;1862:030082.

  22. Shettima AY, Karumi Y, Sodipo OA, Usman H, Tijjani MA. Gas chromatography-mass spectrometry (GC-MS) analysis of bioactive components of ethyl acetate root extract of Guiera senegalensis J F Gmel. J Appl Pharm Sci 2013;3:146-50.

  23. Delazar A, Reid RG, Sarker SD. GC-MS analysis of essential oil of the oleoresin from Pistacia atlantica var mutica. Chem Nat Compounds 2004;40:24–7.

  24. Brophy JJ, Rahmani M, Toia RF, Croft KD, Lassak EV. The volatile oils of Euodia hortensis forma hortensis. Flav Frag J 1985;1:17-20.

  25. Handayani PA, Nurcahyanti H. Ekstraksi minyak atsiri daun zodia (Evodia suaveolens) dengan metode maserasi dan distilasi air. Jurnal Bahan Alam Terbarukan 2015;4:1-7.