Agarwood Resin Research Articles

Growth Performance of Agarwood Resin Producing Gyrinops walla Gaertn under Different Shade Conditions

Gyrinops walla Gaertn is the only agarwood resin producing species naturally growing in Sri Lanka, which thrives well in home gardens and forested areas of the wet zone under different light conditions. Due to the scientific discovery of its ability to produce commercially valuable agarwood resin in this species in 2012, G. walla became popular among many sectors. Since then, growers and investors were searching for necessary information for G. walla plantation establishment and sustainably managing them. Due to lack of such information, the present study aimed at finding the performance of G. walla under different light levels. For this reason, 60 G. walla seedlings were planted in the Rubber Research Institute premises in Agalawatte, Sri Lanka in 2015. After 12 months, six plants were provided with 50% shade, and another six plants were provided with 30% shade. The rest of the plants were kept under full sun. After another 14 months, height and diameter of all plants were measured at monthly intervals. Leaf area and leaf chlorophyll content were measured at three month intervals for six plants in each shade level. Lean nitrogen, phosphorus, potassium, and magnesium contents were analyzed at six month intervals covering both wet and dry periods for six replicates under each shade level. According to the results, height and leaf area of G. walla grown under full sun was significantly lower than those under 50% and 30% shade levels. Stem diameter and leaf chlorophyll contents of G. wall grown under different shade levels were not significant. Further, leaf nitrogen, phosphorus, potassium, and magnesium contents did not significantly vary between wet and dry seasons or between shade levels. The survival rate of the plants after 54 weeks after planting as 88%. The results suggest that, G. walla plantations can be successfully established in the low country wet zone of Sri Lanka, and plants grown initially under shade perform better than those grown under full sun.

The fungal strain promotes rapid agarwood resin production with medicinally accepted agarotetrol level

The fungal strain promotes rapid agarwood resin production with medicinally accepted agarotetrol level

Chemical analysis of different parts from agarwood columns by artificially agarwood-inducing method based on GC–MS and UPLC-TOF-MS

Agarwood is resin-containing wood produced by plants that have been injured. It is widely used in herbal medicine, incense, decorative items, and so on. In this study, we conducted resin area statistical analysis, determined starch particle and reducing sugar contents, and performed multivariate statistical analysis of chemical composition by GC–MS and UPLC-Q-TOF-MS to explore the different components in sections cut from an agarwood column, designated as A1–A4. The results showed that after stimulation by Agar-Bit inducer, the internal phloem parenchyma cells of the column started to form agarwood, and then starch granules were converted into soluble reducing sugars and agarwood resin. Section A1 showed rapid loss of starch granules, resulting in higher contents of reducing sugars and resin. The resin areas of agarwood in the respective sections were different, gradually decreasing on going from A1 to A4. Total numbers of metabolites of 87 and 63 were identified by GC–MS and UPLC-Q-TOF-MS, respectively. Of these, 10 and 16 metabolites with significant differences (variable importance projection >1) were selected through multivariate statistical analysis. These metabolites included chromones, sesquiterpenes, alkanes, and fatty acids. Among them, 6-methoxy-2-(2-phenylethyl)chromone and 6,7-dimethoxy-2-(2-phenylethyl)chromone were significant markers detected by both GC–MS and UPLC-Q-TOF-MS, which may be essential substances responsible for differences in the agarwood-forming capacities of the cut sections. In conclusion, there has been limited research on the different agarwood-forming capacities of agarwood columns. Here, we explored the differences in various sections of agarwood through chemical analysis to provide a more comprehensive and in-depth understanding of its constitution.

Non-Destructive Assessment of Agarwood Resin Formation Status in Aquilaria crassna using Near-Infrared Spectroscopy

Agarwood is a unique and valuable resin synthesized within a few tree species of the tree family Thymelaeaceae primarily as a self-defence mechanism against stress causes such as fungal invasions. Instead, artificial inoculation leads to short-time harvesting practiced in intensive commercial-level cultivations. The extraction of this resin can be done only after the harvesting where accurate estimation of resin yield is crucial for avoiding immature or underdeveloped harvesting. However, the amount of resin accumulated in the trunk cannot be estimated by the outside inspection. As a result, trees are being cut down before they have reached their maximum harvest potential. Therefore, precise detection of fragrant resin formation status inside the tree has become one of the primary challenges in the agarwood production industry. Currently, the methods available are either destructive, inefficient, or inaccurate. As such, this study evaluates the possibility of the use of NIR spectroscopy as a non-destructive and quick detection method for agarwood formations status inside trees in the in-situ mode. Near-infrared (NIR) range (588–1,025 nm) spectra were acquired from resin-formed and non-formed areas from twenty tree trunks of agarwood with five different formation levels. The best agarwood formation areas of logs were assigned as class level 1 and the least was assigned as the class level 5. The spectra were acquired near the opening made for the artificial inoculation process of considered Aquilaria crassna tree trunks. The results of Soft Independent Modelling of Class Analogies showed consistent evidence of differences in the NIR spectral profiles of all levels of agarwood formations. The SIMCA discrimination between the resin developed versus not was achieved with 86% accuracy. The accuracy rates of the degree of the presence of agarwood were obtained as 80%, 100%, 85%, 90%, and 85% respectively, as assigned levels 1 to 5 in the SIMCA models. This research revealed the potential possibility of using NIR technology for the prediction of agarwood resin status inside the agarwood tree in A. crassna before harvesting the tree as a non-destructive and rapid method.
 
 Keywords: NIR spectroscopy, Agarwood, Aquilaria crassna, Non-destructive, Agarwood formation

Investigation of factors affecting the rapid detection of agarwood formation in <em>Aquilaria crassna</em> by near-infrared spectroscopy

Agarwood is a highly valued fragrant resin produced inside a few tree species belonging to the family Thymalaeaceae as a self-defense response to plant stress. The amount of resin developed inside the tree cannot be estimated by outside inspection. Consequently, harvesting trees before they reach their potential yield is a severe drawback to the Agarwood industry. Therefore, developing effective techniques for detecting Agarwood resin status inside the tree species has become a critically important task for the agarwood industry to increase productivity. The present study evaluates the factors affecting Near-Infrared Spectroscopy (NIRS) models when predicting agarwood formation inside A. crassna trunks using NIR spectroscopy. The research used 110 wood specimens obtained from well-grown Agarwood trees in a commercial plantation in Nawimana GS Division, Matara District, Sri Lanka. NIR meter FQA-NIR Gun (588-1100nm) with a custom-made probe was used to acquire NIR reflectance spectra without outside light interference. SIMCA models were built to identify the agar resin-developed wood log areas from the normal wood areas in the tree trunk. SIMCA prediction models were built to investigate three influencing factors, namely present or absent outside tree bark, surface roughness and wood thickness agarwood prediction. Better prediction results were obtained from the bark-removed samples (at the accuracy rates of 97%) to the bark present (85%), smooth wood surfaces (98%) to the rough surface (90%) and 2mm thickness (98%) to the other thickness. The most effective wavelength for the separation of Agarwood present and absent samples was located at 978 nm of NIR. The study has demonstrated the potential possibility of using NIR spectroscopy to identify the agarwood formation in A. crassna in non-destructive and rapid mode.

Barrier zone formation and development in the stems of Aquilaria sinensis (Thymelaeaceae) and the effect on agarwood formation

Summary Barrier and reaction zones are core parts of compartmentalization in trees. However, little is known about the relationship between them. Here, agarwood and barrier zone formation in Aquilaria sinensis trees were studied for the first time using compartmentalization theory. The main results were as follows: (1) Wounding methods had a very important influence on the occurrence of barrier zones in A. sinensis. (2) The barrier zone development process was divided into three phases based on the morphological features. (3) The agarwood resin mainly accreted during the barrier zone lag and division phases; then, after the barrier zone entered the differentiation phase, the resin no longer accreted and even tended to degrade. (4) In the application of the whole-tree agarwood-inducing technique, the barrier zone generally began to appear 10 months after the technique treatment, and the barrier zones were more severe in the lower part of the stem. (5) Considering the agarwood yield, agarwood resin filling degree and ethanol-soluble extract content, the serious barrier zone indeed reduced the agarwood productivity. This study is important for revealing the agarwood and barrier zone formation mechanism in Aquilaria trees. It also provides a good research model for considering the high economic value of agarwood when studying compartmentalization.

Recent Advancement in Agarwood Induction Technology: A Comprehensive Review for the Transformation of Artificial Agar Resin Induction Methods

Agarwood is a high valued resinous wood containing aromatic constituents of sesquiterpenes and phenyl-ethylchromones. They are typically found in the wood tissue of Thymalaeaceae family trees such as Aquilaria and Gyrinops species once they have been physically or chemically damaged or diseased by microbial pathogens. However, the natural occurrence based agarwood production is inadequate to fulfill the worldwide market demand as it never reach to the potential yield. As a result, recent advancements in artificial agarwood induction technology have led to the efficient production of agarwood resin, surpassing conventional methods. These technologies include mechanical, biological, and chemical methods, as well as combinations of these techniques such as fungal inoculation, nailing, drilling, partly trunk pruning, aeration method, agar-wit, C-A kits, Agar-sit, Bottle dipping, ChemJet, Pinhole infusion, Automated transfusion, Agar-Bit and Bamboo stick method. While interdisciplinary approaches have pros and cons, chemical inducers have shown rapid invasion inside particular tree species trunks to produce superior agarwood resin at consistent rate. Therefore, diverse forms of inducers can be utilized to develop this finest fragrance resinous wood in cultivated trees within a short period of time compared to the natural approach and almost similar in the quality compared to the natural agarwood formations. The objective of this review paper was to presents a comprehensive collection of agarwood resin inducing methods and their potential to enhance the total production and the quality of agarwood in the final harvest. The application of these techniques has significant implications to the agarwood industry as it seeks to meet the growing global demand for this highly sought-after and valuable product leading to a high profitable business. Therefore, this review article serves as a valuable collection of resource for the researchers and industry professionals who are ambitious to develop their agarwood industry to a new level.

Endophyte fungi from Agarwood and Mangrove and their applications

Mangroves are woody trees or shrubs salt marsh halophytes that encompass over 75% of the tropical coastline on the planet. Mangroves are characterized by their sophisticated morphological and physiological adaptations as well as their rich synthesis of biologically active secondary metabolites. They also contribute in decreasing CO2 levels in the atmosphere through photosynthesis. On the other hand, agarwood, is an aromatic perfumed precious heartwood of diseased timber of Aquilaria and Gyrinops trees that is used centuries ago till now as incense in religious ceremonies, raw material in the perfume industry and as a medicine. Agarwood formation is usually induced by wounding trunks and branches of Aquilaria trees. Production of natural agarwood resin is inefficient and don’t meet global demand, so different methods have been applied to stimulate biological agarwood induction, and using endophytic fungi is one of the methods. Hence, this review highlights the importance of endophytic fungi in mangroves and agarwood, their existence and some of their important biological activities.

The anatomical structure of the root, stem, and branch of Gyrinops versteegii trees from different growing sites

Abstract. Adimahavira A, Lukmandaru G, Pujiharti R, Prastiwi FW, Nugroho WD. 2023. The anatomical structure of the root, stem, and branch of Gyrinops versteegii trees from different growing sites. Biodiversitas 24: 4590-4598. Gyrinops versteegii (Gilg) Domke is an agarwood (Gaharu)-producing species with a unique color and very expensive fragrance. Even so, studies on the effects of growing sites on the wood anatomical structure of several parts of G. versteegii have not been thoroughly investigated. Therefore, this research aims to investigate the anatomical structure of G. versteegii on the tree's root, stem, and branch at two different growing sites. Three five-year-old G. versteegii trees from Pacitan and Klaten Districts (Indonesia) were used for the experiment and were harvested from the root, stem, and branch. The wood samples were then put in a glutaraldehyde solution for 24 hours and later in an alcohol solution. The samples were sliced in microtome (15-20 µm) and observed in the transverse, tangential, and radial sections. The results showed that the anatomical characteristics, such as vessel diameter, ray height, ray frequency, interxylary phloem proportion, and frequency differed significantly between growing sites. Furthermore, fiber length, vessel diameter, ray height, ray frequency, and interxylary phloem proportion significantly differed among plant parts. In addition, the branch part has the highest interxylary phloem value, indicating the possibility of a high agarwood resin deposit when agarwood stimulation is applied.

Genome-wide investigation of Cytochrome P450 superfamily of Aquilaria agallocha: Association with terpenoids and phenylpropanoids biosynthesis

Agarwood is a dark resinous wood, produced when Aquilaria tree responds to wounding and microbial infection resulting in the accumulation of fragrant metabolites. Sesquiterpenoids and 2-(2-phenylethyl) chromones are the major phytochemicals in agarwood and Cytochrome P450s (CYPs) are one of the important enzymes in the biosynthesis of these fragrant chemicals. Thus, understanding the repertoire of CYP superfamily in Aquilaria can not only give insights into the fundamentals of agarwood formation, but can also provide a tool for the overproduction of the aroma chemicals. Therefore, current study was designed to investigate CYPs of an agarwood producing plant, Aquilaria agallocha. We identified 136 CYP genes from A. agallocha genome (AaCYPs) and classified them into 8 clans and 38 families. The promoter regions had stress and hormone-related cis-regulatory elements which indicate their participation in the stress response. Duplication and synteny analysis revealed segmental and tandem duplicated and evolutionary related CYP members in other plants. Potential members involved in the biosynthesis of sesquiterpenoids and phenylpropanoids were identified and found to be upregulated in methyl jasmonate-induced callus and infected Aquilaria trees by real-time quantitative PCR analyses. This study highlights the possible involvement of AaCYPs in agarwood resin development and their complex regulation during stress exposure.

The induction techniques of resinous agarwood formation: A review

Agarwood is an aromatic, resinous plant that contains volatile, fragrant sesquiterpenes and phenylethylchromones found in wood tissue near damaged or diseased Aquilaria sp. However, natural agarwood production from young, healthy plants is insufficient to meet global market demand. Therefore, this article evaluates the impact of agarwood production methods on the success of the agarwood industry. Recent advances in artificial agarwood induction technology, including mechanical, bio-inoculants, chemical inducers, combinations of biological and chemical inducers, and in vitro plant tissue culture, have resulted in efficiently induced agarwood resin compared with conventional methods. Although interdisciplinary approaches have many advantages and disadvantages, it is encouraging that chemical inducers have a wide distribution in the trunk of Aquilaria sp. Also, able to consistently produce high quality agarwood resin. It can be concluded that different forms of inducers can be used to develop excellent quality agarwood in young plantation trees, equivalent to wild agarwood.

Presence of Actinomycetes in agarwood tissues of Aquilaria crassna: A preliminary study

Agarwood is a valuable resin produced inside certain tree species of the family Thymalaeaceae distributed in the Asian region. Agarwood production occurs as a defense mechanism when the trees are under physical or biological stresses. However, the formation of agarwood resins in significant extractable quantities due to natural stress conditions is rare; therefore, the farmers use various methods to induce its formation artificially. Certain fungal species such as Fusarium and Aspergillus become more popular among them to produce high-quality agarwood. However, studies are rare on using other microbial organisms such as Actinomycetes, which exhibit properties of both bacteria and fungi. Among the agarwood-producing species, Aquilaria crassna is one of Asia's most commonly planted species for agarwood production. This species was introduced to Sri Lanka in 2012 for mid and lower elevations of the wet zone. Due to the lack of studies on agarwood resin formation by non-fungal microbial methods, the present study attempted to identify the presence of Actinomycetes species in agarwood resinous tissues of A. crassna. Agarwood resinous tissue samples were collected from four plantations in the wet zone of the country. Surface sterilized, small sized tissues were placed on starch casein agar medium and incubated at room temperature for ten days. Using the morphological and microscopic characteristics, it was possible to identify Nocardia, Psuedonocardia, and three Streptomyces species with varying abundance. The species level should be confirmed using molecular analysis, and their potential for agarwood resin formation inducement should be tested by re-inoculating to the healthy A. crassna trees.

Programmed cell death might involve in progress of wounding induced agarwood formation in stems of Aquilaria sinensis.

Agarwood, a non-timber fragrant wood, is derived from wounded Aquilaria trees (Thymelaeaceae) and is widely used in traditional medicine, incense and perfume. Agarwood-like substances and programmed cell death (PCD) can be induced by wounding signals in the suspension cells and aerial roots of Aquilaria sinensis. In this study, the dynamic process of wound-induced agarwood formation in stems of A. sinensis was observed, and the occurrence of PCD was synchronously detected using techniques such as 4',6-diamidino-2-phenylindole and dUTP nick-end labeling staining. The results showed that the wounding was a induce signal for agarwood resin formation, meanwhile might induce PCD. Interxylary phloem and xylem ray were the main sites of agarwood resin formation and PCD occurrence. There might be a relationship between the spatiotemporal pattern of PCD and agarwood resin formation: more severe PCD corresponded to a higher rate of resin formation but a shorter resin formation time; conversely, slower PCD progression corresponded to a lower rate of resin formation but a longer resin formation time. Our findings are the first to demonstrate that PCD might occur in the process of wound-induced agarwood formation at the tree level, and the spatiotemporal pattern is closely related to the formation of agarwood resin. This study provides valuable insight for further studies on the relationship between PCD and agarwood formation. HIGHLIGHTS: Programmed cell death (PCD) might occur in the process of wound-induced agarwood formation at the tree level. Interxylary phloem and xylem ray were the main sites of agarwood resin formation and PCD occurrence. Spatiotemporal pattern of PCD might have a strong impact on agarwood resin formation.

Production of Volatile Compounds by a Variety of Fungi in Artificially Inoculated and Naturally Infected Aquilaria malaccensis

Aquilaria malaccensis, the resinous agarwood, is highly valued in the perfumery and medicinal industry. The formation of fragrant agarwood resin inconsistently by various fungi is still not clearly understood. The current study investigatedthe agarwood quality and fungal diversity in artificially inoculated and naturally infected A. malaccensis. The chemical analysis of volatile compounds of agarwood was performed using the Solid Phase Micro Extraction (SPME) method, and the identification of fungi was made through a morphological observation using a light microscope. Gas chromatography analysis revealed the presence of essential compounds related to high-quality agarwood, such as 4-phenyl-2-butanone, β-selinene, α-bulnesene, and agarospirol in both artificially inoculated and naturally infected agarwood but with some differences in the abundance. Further studies onthe fungi associated with agarwood volatile compounds formation showed a total of ten fungal group isolates, which were identified based on morphological and molecular studies. The study revealed that agarwood from both artificial and natural sources were naturally infected with Fusarium, Botryosphaeria, Aspergillus, Schizophyllum, Phanerochaete, Lasiodiplodia, Polyporales, and Ceriporia species. This study has offered a potential opportunity to research further the promising development of fungal strains for artificial inducement of high-quality agarwood formation from A. malaccensis trees.

The effect of crown pruning and induction of Acremonium sp. on agarwood formation in Gyrinops caudata in West Papua, Indonesia

Abstract. Auri A, Faridah E, Sumardi, Hardiwinoto S. 2021. The effect of crown pruning and induction of Acremonium sp. on agarwood formation in Gyrinops caudata in West Papua, Indonesia. Biodiversitas 22: 2604-2611. Agarwood-producing trees have been planted by the community, but have currently not produced agarwood resin. This research examines the crown pruning effect and fungi inoculation effectivity on the formation of agarwood resin in Gyrinops caudata. The environmental modification was also ascertained based on the real environmental situation in natural forests. Furthermore, it was assumed that environmental conditions significantly influenced the growth, G. caudata tree association, and fungi infection. This method involved the tree crown cover modification. Tree crown was categorized into three different classes, i.e. dense crown (no pruning); moderate (pruning 25%), and sparse (pruning 50%). The results showed that inoculating fungi of Acremonium sp. to infection coverage area after three months produced the best results, due to Fcount 79.671. However, crown prunings obtained through heavy, moderate and non-pruning did not show any effect on stem infection spread. In the sixth month, it was observed that the fungi inoculation factor of Acremonium sp. had a significant effect on infection spread formation by Fcount of 168.894. The change in wood internal tissue also had a highly significant effect as depicted by Fcount 461.516. The fragrance level treatment also showed a considerable effect with Fcount 290.385.

Same useful fixative from resin in Vietnam

Today, fixatives from natural raw materials are gradually replacing fixatives synthesized from chemicals. The arm of this research is the study of same useful resin in Vietnam for fixative substance in blending fragrance. We obtained Benzoin resin from Styrax tonkenensis Pierre plant in Ha Giang province, Canarium resin from Canarium Album L. in Dak Nong district, Dak Lak province, and Agarwood pulp of Aquilaria crassna plant in Binh Thuan province Vietnam. The material used in the experiment for the natural fragrance was taken from the project of Vietnam essential oils and related natural products. The method of this process is resin extraction by volatile solvents. The resin is dissolved in alcohol 96% and the distilled alcohol is removed to obtain absolute. The method of assessment of product quality in this study is using the olfactory to assess the odour of samples over time. Benzoin resin, Canarium resin, and Agarwood resin of Vietnam are useful fixatives in blending fragrance. The fixative ability of Benzoin resin absolute is not equal to the ability of Agarwood resin absolute but better than the ability of Canarium resin absolute. Through research and experiment, we can see Benzoin resin, Canarium resin and Agarwood resin are precious. They can be used as a good fixative in aromatherapy. This is a natural resin, a kind of resource available in Vietnam. Therefore, it is recommended for further research, exploitation, and effective use of this resource.

Production of Agarwood Resin in Aquilaria beccariana Using Inducement Technology

Agarwood is a type of resin impregnated wood produced from the wounded Aquilaria trees. This agarwood gives a pleasant fragrant when it is burned. It becomes high-priced and increase demanded in the world due to the depletion of wild agarwood in the forest caused by illegal poaching activities. Agarwood resin can only be produced by injuring caused by lightning or wounded by animals under natural conditions. However, the natural process of resin accumulation is uncertain and time-consuming. Therefore, we developed an agarwood inducement technique that served as the alternative way to induce the agarwood formation in a short time. Three inducement techniques, including the injecting method, knocking method and combination of injecting & knocking method were applied to induce resin formation. In this study, we evaluated the technique for producing agarwood in species Aquilaria beccariana, which is native and only can be found in Borneo Malaysia. For A. beccariana trees treated with the inducement technique, resin formed and spread throughout the cell in the trunk. The evaluation results showed that the agarwood yield per tree reached around 5 to 7 kilogram. Furthermore, this agarwood derived from the induction was found to have a similar quality with the wild agarwood. This indicates the inducement technology had successfully produced agarwood resin in A. beccariana with a grade similar to the wild agarwood.

A REVIEW: AQUILARIA SPECIES AS POTENTIAL OF PHARMACOLOGICAL

One of the plants that is rich in benefits is from the genus Aquilaria spp. which is known to be able to produce agarwood resin. This genus includes 47 species, of which the four main species are Aquilaria malaccensis, Aquilaria subintegra, Aquilaria crassna and Aquilaria sinensis. Aquilaria species are known to have a wide spectrum of pharmacological activity and have been reported in many studies, including activity, anticancer, anti-inflammatory, AChE inhibitor (Acetylcholinesterase), anti-proliferative, and antidiabetic. The aim of this review is to expand information on the pharmacological activity of various Aquilaria species that can support future studies in the medical field of medicine.

Study on Soil Properties towards Formation of High Quality Agarwood Resin in <i>Aquilaria crassna</i>

Agarwood has been used for its fragrance and medicinal properties in Asian culture for centuries. In recent years, agarwood gains its popularity in the west due to its usage in perfume formulation. Unfortunately its supply not meets the augmentation market demand. This is mostly because of depleting nature agarwood reservoir and lack of induction technique that can produce agarwood consistently in the plantation. In this study, we look into a case where artificial inducing technique successfully produced high quality agarwood. To assure its quality, agarwood chip was analysed by Gas Chromatography for its chemical profiles. Discovered compounds were identified as sesquiterpene group which also had been characterized as major agarwood compound listed on previous studies. Few compounds that are detected such as δ-cadinene (0.20%), jinkoh-eremol (22.09%), epi-α-cadinol (4.74%), agarospirol (3.75%) and others. Soil condition that contributes into this agarwood formation; soil analysis on physical properties, chemical properties and nutrients content of the soil have been analyzed. Based on the findings, soil condition is an important factor to successfully induced high quality agarwood.

Evaluation of a Method of Separating Agarwood Absolute from Aquilaria crassna Pulp by Soxhlet and Soaking for Fixative Substance in Blending Fragrance

Agarwood resin is a precious material that has been widely used in industry and life. The aim of this study is the evaluation of a method of separating Aquilaria crassna wood pulp with ethanol by Soxhlet and solvent immersion method for fixative substance in blending fragrance. Agarwood pulp of Aquilaria crassna tree is grown in Binh Thuan province, Vietnam. Our research team has extracted the Agarwood absolute F.1 and F.2 from Aquilaria crassna by Soxhlet extraction and soaking method. The fragrance is diluted 10 times by odorless solvent diethyl phthalate (DEP), then use paper and olfactory of the nose to evaluate the aroma. The yield of absolute Agarwood extracted by the method of Soxhlet is 5.56% weight. The yield of Agarwood absolute extracted by the method of soaking is 4.58% weight. Agarwood absolute F.1 is a fixative substance, which is capable of storing fragrance better than Agarwood absolute F.2. The scent of fragrance Fr.F.2 using Agarwood absolute F.2 is more like a natural scent than fragrance Fr.F.1 with fixative F.1. Separating Agarwood absolute F.1 from Aquilaria crassna pulp by Soxhlet provided yield and fixative capabilities higher than separating Agarwood absolute F.2 from Aquilaria crassna pulp by soaking. The scent of fragrance Fr.F.2 using Agarwood absolute F.2 is more like a natural scent than fragrance Fr.F.1 with Agarwood absolute F.1.