The Effect of Enzymatic Pre-Treatment in Agarwood Oil Extraction

Astri Fadhilah Abdul Rahim, Atikah Kadri, Nor Masliyah Harun


Agarwood oil was extracted from a pre-treated agarwood chip soaked in a specific enzyme concentration of 1 %, 3 % and 5 % respectively. The effect of soaking time was also investigated by applying soaking period in the range of 3 to 14 days. Hydrodistillation was carried out using the Clevenger-type apparatus to obtain the essential oil of agarwood chips. Subsequently the quality of the oil and the identification of its oil compounds were analyzed using GC-MS. The morphology of agarwood chips prior to hydrodistillation was characterized using scanning electron microscope (SEM). The SEM results of enzymatic pre-treated sample showed an obvious change in the cell morphology as compared to the sample with standard soaking of 6 days. The highest yield of oil with the amount of 0.125 % is produced from the sample pre-treated with 3 % enzyme concentration as compared to 0.065 % of oil yield from the non-pre-treated sample employing similar soaking days. The GC-MS analysis revealed a total of 335 compounds from all samples of agarwood oil and similar compounds were found in the standard soaking and enzymatic pre-treated samples. Caryophyllene, Gurjunene, and Alloaromadendrene that contributes to the unique odour of agarwood oil are found mostly in all samples. Thus, the enzymatic pre-treatment indicates a comparable yield and oil quality and it is achievable even with a reduced soaking time of 6 days as compared to standard 14 days.


agarwood; essential oil; enzymatic pre-treatment; GC-MS; hydrodistillation extraction.

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S. Najafian, “Storage conditions affect the essential oil composition of cultivated Balm Mint Herb (Lamiaceae) in Iran,” Ind. Crops Prod., vol. 52, pp. 575–581, 2014.

D.-Y. Zhang, X.-H. Yao, M.-H. Duan, F.-Y. Wei, G.-H. Wu, and L. Li, “Variation of essential oil content and antioxidant activity of Lonicera species in different sites of China,” Ind. Crops Prod., vol. 77, pp. 772–779, 2015.

D. P. De Sousa, “Bioactive Essential Oils and Cancer,” Bioact. Essent. Oils Cancer, pp. 1–292, 2015.

P. Costa, B. Medronho, S. Gonçalves, and A. Romano, “Cyclodextrins enhance the antioxidant activity of essential oils from three lamiaceae species,” Ind. Crops Prod., vol. 70, pp. 341–346, 2015.

M. R. Haji Che Daud and H. Hamdan, “Production and Chemical Compounds of Gaharu Extract by Using Supercritical Fluid Extraction (SFE) of Unsoaked Gaharu at Different Pressure,” Adv. Mater. Res., vol. 1115, pp. 337–340, 2015.

S. K. N. Afiffudden, H. Alwi, and K. H. K. Hamid, “Determination of 4’-hydroxyacetanilide in leaves extract of Aquilaria malaccencis by high pressure liquid chromatograph,” Procedia - Soc. Behav. Sci., vol. 195, pp. 2726–2733, 2015.

I. H. Burkill, W. Birtwistle, F. William Foxworthy, J. B. Scrivenor, and J. Gilbert Watson, A dictionary of the economic products of the Malay Peninsula. Published on behalf of the governments of Malaysia and Singapore by the Ministry of Agriculture and cooperatives. 1966.

T. Nakanishi et al., “Three Fragrant Sesquiterpenes of Agarwood*,” vol. 23, no. 9, pp. 2066–2067, 1984.

J. Alam et al., “An insight of pharmacognostic study and phytopharmacology of Aquilaria agallocha,” J. Appl. Pharm. Sci., vol. 5, no. 8, pp. 173–181, 2015.

D. T. Ahmaed, M. Mahmood, A. M. Masaad, and S. N. Tajuddin, “Investigation of Agarwood Compounds in Aquilaria malaccensis & Aquilaria Rostrata Chipwood by Using Solid Phase Microextraction,” Biomed. J. Sci. Tech. Res., vol. 1, no. 6, pp. 1–8, 2017.

M. Mahfud, A. Chandra K. F., L. Qadariyah, and P. Prihatini, “Distillation Assisted by Microwave for Extracting Essential Oil from Java Cananga Flowers,” Mod. Appl. Sci., vol. 9, no. 7, pp. 199–205, 2015.

M.-T. T. Golmakani and M. Moayyedi, “Comparison of microwave-assisted hydrodistillation and solvent-less microwave extraction of essential oil from dry and fresh Citruslimon (Eureka variety) peel,” J. Essent. Oil Res., vol. 28, no. 4, pp. 272–282, 2016.

M. Puri, D. Sharma, and C. J. Barrow, “Enzyme-assisted extraction of bioactives from plants,” Trends Biotechnol., vol. 30, no. 1, pp. 37–44, 2012.

D. Rodrigues et al., “Impact of enzyme- and ultrasound-assisted extraction methods on biological properties of red, brown, and green seaweeds from the Central West Coast of Portugal,” J. Agric. Food Chem., vol. 63, no. 12, pp. 3177–3188, 2015.

K. C. Baby and T. V. Ranganathan, “Effect of enzyme pre-treatment on extraction yield and quality of cardamom ( Elettaria cardamomum maton .) Volatile oil,” Ind. Crop. Prod., vol. 89, pp. 200–206, 2016.

C. P. Passos, S. Yilmaz, C. M. Silva, and M. A. Coimbra, “Enhancement of grape seed oil extraction using a cell wall degrading enzyme cocktail,” Food Chem., vol. 115, no. 1, pp. 48–53, 2009.

J. Jiao et al., “Enzyme-assisted microwave hydro-distillation essential oil from Fructus forsythia, chemical constituents, and its antimicrobial and antioxidant activities,” vol. 134, pp. 235–243, 2012.

J. Chandran, K. Parukutty, P. Amma, and N. Menon, “Effect of Enzyme Assisted Extraction on Quality and Yield of Volatile Oil from Black Pepper and Cardamom,” vol. 21, no. 6, pp. 1611–1617, 2012.

A. Boulila et al., “Enzyme-assisted extraction of bioactive compounds from bay leaves (Laurus nobilis L.),” Ind. Crops Prod., vol. 74, pp. 485–493, 2015.

N. Yoswathana, “Extraction of agarwood (Aquilaria crassna) oil by using supercritical carbon dioxide extraction and enzyme pretreatment on hydrodistillation,” J. Food, Agric. Environ., vol. 11, no. 2, pp. 1055–1059, 2013.

V. D. Dimaki, G. Iatrou, and F. N. Lamari, “Effect of acidic and enzymatic pretreatment on the analysis of mountain tea ( Sideritis spp.) volatiles via distillation and ultrasound-assisted extraction,” J. Chromatogr. A, 2017.

V. A. Jok, N. C. Radzi, and K. H. K. Hamid, “Agarwood Oil Yield As A Result of Changes in Cell Morphology Due To Soaking Process,” Procedia - Soc. Behav. Sci., vol. 195, pp. 2443–2450, 2015.

M. Bayram and A. Kaya, “Changes in properties of soaking water during production of soy-bulgur,” J. Food Eng., vol. 61, pp. 221–230, 2004.

V. A. Jok, M. Arif, F. Zainudin, N. C. Radzi, and K. H. Ku, “Compounds Identification in Agar Wood Soaking Water by Using GC-MS,” vol. 7, no. 5, pp. 340–343, 2016.

Y. Chen, R. R. Sharma-Shivappa, D. Keshwani, and C. Chen, “Potential of Agricultural Residues and Hay for Bioethanol Production,” pp. 276–290, 2007.

J. S. Van Dyk and B. I. Pletschke, “A review of lignocellulose bioconversion using enzymatic hydrolysis and synergistic cooperation between enzymes — Factors affecting enzymes, conversion and synergy,” Biotechnol. Adv., vol. 30, no. 6, pp. 1458–1480, 2012.

P. Pripdeevech, W. Khummueng, and S.-K. Park, “Identification of Odor-active Components of Agarwood Essential Oils from Thailand by Solid Phase Microextraction-GC/MS and GC-O,” J. Essent. Oil Res., vol. 23, no. 4, pp. 46–53, 2011.

H. Chen, Y. Yang, J. Xue, J. Wei, Z. Zhang, and H. Chen, “Comparison of compositions and antimicrobial activities of essential oils from chemically stimulated agarwood, wild agarwood and healthy Aquilaria sinensis (Lour.) Gilg trees,” Molecules, vol. 16, no. 6, pp. 4884–4896, 2011.

Y. Z. .-Y. Hashim, N. I. Ismail, and P. Abbas, “Analysis of Chemical Compounds of Agarwood Oil From Different Species By Gas Chromatography Mass Spectrometry ( Gems ),” IIUM Eng. J., vol. 15, no. 1, pp. 55–60, 2014.

T. Acree and H. Arn, “Flavornet and human odor space, gas chromatography - olfactometry (GCO) of natural products,” 1984. [Online]. Available:

S. N. Tajuddin, C. M. Aizal, and M. M. Yusoff, “Resolution of Complex Sesquiterpene Hydrocarbons in Aquilaria malaccensis Volatile Oils Using Gas Chromatography Technique,” in Agarwood: Science behind the Fragrance, R. MOHAMED, and Ed. Singapore: Springer Singapore, pp. 103–124, 2016.

K. Hosni, I. Hassen, H. Chaâbane, M. Jemli, and S. Dallali, “Enzyme-assisted extraction of essential oils from thyme (Thymus capitatus L.) and rosemary ( Rosmarinus officinalis L .): Impact on yield , chemical composition and antimicrobial activity,” Ind. Crop. Prod., vol. 47, pp. 291–299, 2013.

N. A. Mat Yusoff, S. N. Tajuddin, A. Hisyam, and N. A. Mohd Omar, “Agarwood Essential Oil : Study on Optimum Parameter and Chemical Compounds of Hydrodistillation Extraction,” J. Appl. Sci. Agric., vol. 10, no. 5, pp. 1–5, 2015.

N. F. K and K. H. Ku Hamid, “Effects of soaking on yield and quality of agarwood oil,” J. Trop. For. Sci., vol. 24, no. 4, pp. 557–564, 2012.



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