Phytochemical and tannin content in two

species of agarwood leaves from Mandailing

Natal Regency North Sumatera Province
Cite as: AIP Conference Proceedings 2049, 030009 (2018); https://doi.org/10.1063/1.5082510
Published Online: 14 December 2018

Ridwanti Batubara, T. Ismanelly Hanum, and Surjanto

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AIP Conference Proceedings 2049, 030009 (2018); https://doi.org/10.1063/1.5082510 2049, 030009

© 2018 Author(s).
 Phytochemical and Tannin Content in Two Species of
Agarwood Leaves From Mandailing Natal Regency North
Sumatera Province
, 2, a)
Ridwanti Batubara1 , T Ismanelly Hanum3, 4, b) and Surjanto3, c)

1
Faculty of Forestry, Universitas Sumatera Utara, Jl. Tridharma Ujung No.1, Medan 20155, North Sumatera,
Indonesia
2
Center of Excellence for Natural Resources Based Technology, Mangrove and Bio-Resources Grup, Universitas
Sumatera Utara, Medan 20155, North Sumatra, Indonesia
3
Faculty of Pharmacy, Universitas Sumatera Utara, Jl. Tridharma Ujung No.4, Medan 20155, North Sumatera,
Indonesia
4
Nanomedicine Center of Innovation, Universitas Sumatera Utara, Medan.
a)
Corresponding author: ridwantibb@yahoo.com
b)
isma_nelly@usu.ac.id
c)
yanto_juni@ymail.com

Abstract

Agarwood (Gaharu) leaves have been used as raw material for tea. This study aims to identify the secondary metabollites
and tannin content from two species agarwood (Aquilaria malaccensis Lamk and Wikstroemia tenuiramis Miq) sited in
Mandailing Natal Regency. The leaves were extracted with ethanol for phytochemical and tannin content test. The results
showed that agarwood leaves were positive for alkaloids, flavonoids, steroids/triterpenoids, and tannin. The tannin content
of W. tenuiramis was 4,95 % and A. malaccensis was 5,62 % respectively.
Key words: Agarwood leaves, species, phytochemical, tannin.

INTRODUCTION
The distribution of trees that can produce gaharu in Indonesia are Sumatra, Java, Kalimantan, Sulawesi,
Maluku, Papua and Nusa Tenggara. Agarwood is a resin obtained from microbial infection of trees from
Thymeleacea, Leguminoceae and Euforbiaceae families. Gaharu grows naturally in nature and endemic in North
Sumatra especially in Mandailing Natal Regency. The experiment of agarwood leaves utilization has been done
based on chemical compound content of alkaloids, tannins, saponins, flavonoids and terpenoids [1]. The leaves are
also used as a brew drink that acts as an antioxidant. Exploration results conducted found 2 species of agarwood in
Mandailing Natal Regency, Aquilaria malaccensis Lamk and Wikstroemia tenuiramis Miq, that still same with
agarwood-producing plants and its distribution in Indonesia [2].
Flavonoids, terpenoids and phenol compounds are estimated to have activity as anti free-radical
(antioxidants) [3]. Natural antioxidants are scattered in some parts of plant, such as in wood, bark, roots, fruits,
flowers, seeds, and leaves [4]. Gaharu leaves from Langkat contain a sturdy antioxidant and potencial as raw
material of tea which is rich in antioxidant and have value in more than 3 with enough prefer category in hedonic
test of color, flavor and aroma [5].
Pursuant to that matter hence we conducted a research which aims to see chemical content and tannin
content from two types of gaharu leaves growing in Mandailing Natal. Gaharu leaves will be used as raw materials
of tea rich in antioxidants.

The 3rd International Seminar on Chemistry

AIP Conf. Proc. 2049, 030009-1–030009-5; https://doi.org/10.1063/1.5082510
Published by AIP Publishing. 978-0-7354-1775-5/$30.00

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 RESEARCH METHODS
Location
The research was done from February to May 2018. The sampling site was conducted in Laru Village,
Tambangan District, Mandailing Natal Regency, and Siantona Village, Sorik Marapi Valley District, Mandailing
Natal Regency. For phytochemical screening and tannin testing conducted at Pharmacognosi Laboratory and
Research Laboratory, Faculty of Pharmacy, University of Sumatera Utara.
Procedure
1. Sample Collection
Sampling is done purposively without comparing from other regions. Gaharu leaves, A. malaccensis Lamk.
from Laru village which grow naturally and W. tenuiramis Miq. from Siantona village was used in this research.
2. Raw Material Preparation
Gaharu leaves were cleaned of dirt with flowing water, then distribute on parchment paper until the water
was absorbed. The plant materials were done by unnatural drying dried in the drying cupboard to dry and brittle the
leaves with temperature 40°C-50°C. The purpose was to get the simplicia that was not easily damaged, so it can be
stored for a long time. The dried leaves were grinded using a blender and placed shielded from the sun before
extraction and testing activities.
3. Determination of Water Content
The determination of water content was done by an Azeotropy method (Toluen Distillation) [6].
4. Phytochemical Screening
Phytochemical screening is a qualitative chemical examination of biologically active compounds presenting
within simplicia and plant extracts, organic compounds. Therefore, screening is primarily intended for groups of
organic compounds such as alkaloids, glycosides, flavonoids, steroids/terpenoids, tannins and saponins.
5. Preparation of Ethanol Extract of The Leaves (A. malaccensis Lamk)
The preparation of the extract was carried out maceration with 96% ethanol solvent, as much as 200 g of
simplicia (dust) inserted into a glass vessel, poured with 1500 ml of 96% ethanol, covered the glass, left for 5 days
shielded from light and occasionally stirred. After 5 days, the mixture was filtered. The dregs were washed with
96% ethanol sufficiently to obtain 2000 ml, then removed in a closed vessel and left in place shielded from light for
2 days, then filtered. The maserate was concentrated using a rotary evaporator device at 40 °C until a concentrated
maserate was obtained and then dried using a freeze dryer to obtain a dry extract [6].
6. Tannin Content Test
Testing of tannin content using Lowenthal Procter method [6].

RESULTS AND DISCUSSIONS

Determination of Simplicia Water Content

Determination of water content should be known because it is closely related to the quality of simplicia.
The determination of water content is useful for presuming the durability or resilience of samples in storage with the
result that relate to the quality of the dried leaves. The determination of water content is made to provide a minimum
limit that can still be tolerated in the simplisia.

TABLE 1. Water Content of Gaharu Leaves

Water
No Gaharu Leaves Location
Content (%)
Laru village, Tambangan district, Mandailing
1 7,66
Natal regency (A. Malaccensis)

Siantona village, Lembah Sorik Marapi district,

2. 5,33
Mandailing Natal regency (W. Tenuiramis)

The water content of gaharu leaves from Laru, A. malaccensis and from Siantona, W. tenuiramis, can be
seen in Table 1. The water content of the two types were relatively similar because of the same process. The
determination of water content was useful to estimate the durability of samples in storage as well as to correct the
yield. Water content of simplicia of natural materials usually must be lower than 10% to avoid the bacterial or fungi

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growth, so the simplicia can be stored for a long time. The water content of the simplicia has fulfilled the standard
requirement of the simplicia water content of not exceeding 10% [6].
High water content level can trigger enzymatic reactions as well as microbial growth, especially fungi that
can cause decaying the chemical compounds in simplicia [7]. The water content of A. malaccensis was relatively
lower than W. tenuiramis derived from the Siantona Valley of Sorapi Marapi village (Table 1), although the
difference was only the decimal number.

Determination of Tannin Content

High-low levels of tannin content were influenced by the amount of extract in water (tea) because if tannin
was dissolved into water, the colloid would form, and appeared a sour and bitter taste on gaharu leaves in tea. Value
of tannin content was seen in Table 2.
Tannin is a folifenol compound that can be complex with proteins forming copolymers. Tannin is present in
leaves, immature fruit. Tannin is an active compound of plants belonging to flovonoid group that has a bitter sense
on food [8].
Tannin compounds are belonging to the flavonoid compound [9]. Tanin is a phenol compound that has a
large molecular weight. Tannin is consist of two types of hydrolyzed tannins and condensed tannins.
Tannin A. malaccensis was 5.62% and W. tenuiramis 4.95%. The content of tannins on 2 types of gaharu
leaves was not much different. Tannins and flavonoids in tea are used in ancient medicine to heal burns and L QVHFW¶V
toxins, because tannins and flavonoids are anti-inflammatory and antiseptic.

TABLE 2. Tanin Content

Tannin
No Gaharu Leaves Location
Content (%)
Laru village, Tambangan district,
1 5,62
Mandailing Natal regency (A. Malaccensis)
Siantona village, Lembah Sorik Marapi
2. district, Mandailing Natal regency (W. 4,95
Tenuiramis)

Tannin on Camellia sinensis tea was 1.21% and tannin content of two types of gaharu leaves was higher
than C. sinensis. Fermentation is a process that resulted in chemical changes in tea caused by enzymes. During the
fermentation process, the oxidation of cells released during milling with oxygen happened, in the presence of an
enzyme that acts as a catalyst [10]. The longer the fermentation done on gaharu leaves, the greater the decrease in
tannin content in gaharu leaves. The decrease of tannin was caused by tannin oxidation with polyphenol enzyme
involvement. The high levels of tannin in 2 types in this study due to the absence of fermentation treatment
performed on the leaves, for example by rolling gaharu leaves.

Phytochemical Screening Results

Gaharu leaves research was conducted to increase the benefits of the tree. Phytochemical screening is a
preliminary study to find out secondary metabolites within plants. Phytochemical screening was conducted on fresh
gaharu leaves simplicia dust to obtain information of secondary metabolite compounds class contained in the leaves.
The results of phytochemical screening can be seen in Table 3. The description of the compounds as follows:
1. Alkaloids
Alkaloid compounds were not present in two species of agarwood leaves. This was indicated by the absence of
sediment or turbidity in the experiment using Meyer, Dragendroff, and Bouchardart reagents.
2. Flavonoids
The presence of flavonoid compound was characterized by the yellow colour in simplicia and red colour on
ethanol extracts on the amyl alcohol layer. Flavonoids have many pigments and present throughout the plants.
The flavonoid compounds were a group of the largest phenol compounds found in nature. The phytochemical test
result showed that two species of agarwood leaves contained flavonoids.
3. Tannin
Tannins are generally defined as polyphenol compounds with a high molecular weight (over 1000) and able to
form complexes with proteins. The examined the simplicial two species of agarwood leaves by the addition of
FeCl3 1% to diluted filtrate. This examination resulted blackish-green colour indicating the presence of tannin.

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4. Saponins
Saponin is a strong surface active compound, foaming when shaken in water and at low concentrations often
causing red blood cell haemolysis. The test result showed that two species of agarwood leaves contained saponin.
3. Steroids/Triterpenoids
The phytochemical test result showed that two species of agarwood leaves contained steroid/triterpenoid. The
appearance of red colour after the addition of acetic acid anhydride and concentrated sulfuric acid showed the
presence of steroids/triterpenoids on agarwood leaves.

The results obtained in Table 3 showed that phytochemical screening test did not have any differences in
chemical compound group content conducted on 2 types of gaharu leaves. The similarity of the compounds content
class due to the samples tested were the same leaves that grew naturally. Besides the two studied, Grynops versteegii
also contains secondary metabolite compounds of flavonoids, terpenoids and phenolic compounds [10]. The results
obtained in the phytochemical test showed that gaharu leaves water extract contained phenolic compounds,
flavonoids, and steroids [11].

TABLE 3. Phytochemical Screening Data

Gaharu Types
Compounds
A. malaccensis from Laru W. tenuiramis from Siantona
Alkaloids - -
Flavonoids + +
Tannins + +
Saponins + +
Triterpenoids + +

Gaharu leaves, A. malaccensis Lamk. and W. tenuiramis Miq., have potency as an antioxidant. Flavonoids
generally have potency as antioxidants. The flavonoid compounds act as free-radical capturer because the hydroxyl
group donates hydrogen to free radical [12]. The compounds are able to neutralize free radicals by offering
electrons, so the unpaired atoms get electron pairs and no longer radicals. Their antioxidant effects are indicated by a
variety of mechanisms, including the ability to scavenge free radicals or to activate antioxidant enzymes and inhibit
oxidation [13] . The ability of flavonoids acting as antioxidants is because of its molecular structures [14]. The
leaves of Calophyllum inophyllum contain flavonoid compounds based on phytochemical tests, structurally derived
from flavone substances according to the flavonoid literature, and certain conjugated aromatic systems, which have
antibacterial, anti-inflammatory, anti-allergic, anti-mutagenic, antiviral, anti-neoplastic, anti-thrombotic and
vasodilatory activity [15].
The result of phytochemical screening obtained can give important information about the chemical
compound contained by gaharu leaves. Studying the chemical compounds will facilitate the determination of usage,
especially in the advanced utilization, treatment. Screening technique can help the phytopharmacological measures
of early selection of the plant examination to prove the presence of certain chemical compounds in the plant and can
be attributed to its biological activity [16].
Cytotoxic activity depending on the dose observed in gaharu fraction shows the potential therapeutic
usefulness of this medicinal plant against cervical cancer [17] . A. malaccensis leaves show the ability to increase
glucose uptake by increasing levels of GLUT4 in skeletal muscle. Further experiment is needed to explore this
gaharu leaves as a strong antidiabetic [18].

CONCLUSIONS AND RECOMMENDATIONS

CONCLUSIONS

The conclusion of this study is that two types of gaharu can be used as raw materials, with air content of less than
10% and tannin content below 6%. Both types of gaharu contain alkaloids, flavonoids, steroids / triterpenoids and
tannins.
RECOMMENDATIONS
Nowadays, gaharu is increasingly scarce but has the potential to be exploited more widely, especially the leaves. We
hope to intensify the cultivation of gaharu further.

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 ACKNOWLEDGEMENT
This research is done by financed by: Directorate of Research and Community Dedication of Directorate General of
Research Strengthening and Development of Ministry of Research, Technology and Higher Education 2018 on the
Fundamental Grant Scheme.

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