Journal of Applied and Industrial Sciences, 2013, 1 (3): 78-88, ISSN: 2328-4595 (PRINT), ISSN: 2328-4609 (ONLINE)

78

Research Article

Characterization of Methanolic Extracts of

Agarwood Leaves

Khalil, A. S.1; Rahim, A. A.2, Taha, K. K.*3, Abdallah, K. B. 4

1

Department of Chemistry and Industrial Chemistry, College of Applied and Industrial Sciences, University of Bahri, Sudan
2
School of Chemical Sciences, Universiti Sains Malaysia (USM), Penang- Malaysia

*3Department of Chemistry, College of Science, Al-Imam Mohammad Ibn Saud Islamic University, P.O. Box : 5701, Riyadh, KSA
4

Department of Chemical Engineering, College of Engineering, Karary University, Sudan

(Received: June 14, 2013; Accepted: August 21, 2013)

Abstract Tropical Agarwood (Aquilaria) is in danger of

extinction in the wild due to illegal logging. Its resin (Gaharu) is
used for the production of highly valued incense throughout Asia.
The methanolic crude extracts of Agarwood leaves before and
after inoculation process extracted by using maceration into
methanol as solvent were screened for Phytoconstituents, the
extracts revealed the presence of alkaloids, tannins, saponins,
flavonoids and terpenoids.
Gas chromatography-mass spectrometry (GC-MS) analysis of the
plant extracts led to the identification of 14 components from the
two types of Agarwood leaves extracts. It is interesting to see here
that all the identified compounds from agarwood leaves
Methanolic extracts contained oxygen and/or -electrons in their
molecules. Hexadecanoic acid was found to be one of the major
compounds in Agarwood leaves methanolic extracts.
Infrared spectroscopic analysis of the extracts of Agarwood
revealed the presence of O-H, C=O, C-H, C-N, C-O, -CH2 and
CH3 bond stretching.
The present study has proved the usefulness of agarwood
tree for medicinal and anti corrosive purposes. The presence of
phytochemicals indicates its potential as a source of useful drugs.
Index Terms Agarwood, Inoculation, Methanolic extract,
Phytoconstituents

I. INTRODUCTION

nowledge of the chemical constituents of plant is

helpful in the discovery of therapeutic agent as well
as new sources of economic materials like oil and gums.
The most important bioactive constituents of these plants
are alkaloids, tannins, flavonoids and phenolic compounds
[1].
Studies revealed that agarwood has remarkable anticancer
activity [2]. The Benzene extracts of the plant have central
nervous system antidepression activities [3]. Rise in
demand for agarwood resulted in irrational cutting of the
tree trunk for extraction of the chemicals. This has resulted
in the tree becoming endangered. Agarwood leaf is a
Corresponding author E mail: kamaltha1012@yahoo.com

promising potential antidiabetic agent [4]. The extracts of this

plant, which contains numerous naturally environmental
organic compounds, may be utilized as eco-friendly natural
corrosion inhibitors for metals or alloys in aggressive medias.
There is an intensive effort underway to develop new plant
origin corrosion inhibitors for metal subjected to various
environmental conditions. These efforts have been motivated
by the desire to replace toxic inhibitors used for mitigation of
corrosion of various metals and alloys in aqueous solutions.
Plants represent a class of interesting source of organic
compounds currently being explored for use in metal corrosion
protection in most systems, as possible replacement of toxic
synthetic inhibitors.
Aquilaria Malaccensis (Agarwood) is one of 15 tree species in
the Indomalesian genus Aquilaria, family Thymelaeaceae, [5]
and class Magnoliopsida. It is a large evergreen tree growing
over 15-30 m tall and 1.5- 2.5 m in diameter, and has white
flowers [6]. The Leaves are 5-11 cm long and 2-4 cm broad.
Aquilaria species have adapted to live in various habitats,
including those that are rocky, sandy or calcareous, welldrained slopes and ridges and land near swamps. They
typically grow between altitudes of 0-850 m, in locations with
average daily temperatures of 20-22 C [7].
Aquilaria Malaccensis (Agarwood) is widely distributed in
south and south-east Asia. There are differing accounts of the
countries in which it occurs. According to Oldfield, S., et al.
(1998) [8], A. Malaccensis is found in 10 countries:
Bangladesh, Bhutan, India, Indonesia, Iran, Malaysia,
Myanmar, Philippines, Singapore and Thailand.
Agarwood, eaglewood, gaharu, aloeswood, oud, oudh,
kanankoh, kyara, jinkoh, chen xiang, tomention and
kalamabak- these are just a few of the names for this tree.
Agarwood is the most valuable wood in the world with higher
prices and demands nowadays. The widely uses of agarwood
in meditation field, essential oil production and etc makes
agarwood one of the precious things on earth.
There is no detail systematic documentation of presence
and type of phytochemicals in agarwood leaves. Hence the
present study aimed at an evaluation of the presence of

Journal of Applied and Industrial Sciences, 2013, 1 (3): 78-88, ISSN: 2328-4595 (PRINT), ISSN: 2328-4609 (ONLINE)
different phytochemicals along with GC-MS and FTIR
investigations of methanol soluble crude extracts obtained
from before and after inoculation leaves of Aquilaria
Malaccensis (Agarwood).
II. MATERIALS AND METHODS
Collection of the Plant Materials
Two types of fresh leaves of agarwood were collected from
(Main land-Penang, Malaysia). One was before the inoculation
process (MOB) and the other type was after the inoculation
process (MOA). Both new and old growth leaves were
avoided, damaged and diseased ones were excluded, only
recently matured leaves were taken.
Preparation of the Plant Materials
The leaves were dried in the shade for 7 days at room
temperature (28 2C) and ground to a fine powder using
Grinder IKA-WERKE, IKA MF10 Machine and sieved
through a 0.25 m mesh. The powder samples were kept at
room temperature in a covered glass containers to protect them
from humidity and light prior to extraction.
Extraction of Agarwood Leaves Extracts
250g dried powder leaves of each A. Malaccensis (Agarwood)
type
(before and after inoculation) were exhaustively
extracted by macerated in 1.5 L methanol solvent for 2 days at
room temperature (282C). The solvent-containing extract
was then decanted and filtered by vacuum filtration (GAST,
DOA-P504-BN, USA). The extraction of the ground leaves
were further repeated (twice) with methanol (1 L each time).
The filtrate from each extraction was combined and the excess
solvent was evaporated under reduced pressure at 40C using a
rotary
evaporator
(Heidolph-instruments,
Rotavapor,
Germany) to give concentrated crude methanolic extracts,
dried in oven at 50C to give dark green extracts. The weights
of all the extracts were measured after solvent evaporation and
then kept into a glass container prior to use.
The above mentioned processes were done for each type of
leaves separately.
Preliminary Phytochemical Analysis of the Agarwood
Methanolic Crude Extracts
Extracts were tested for the presence of active principles such
as alkaloids, flavonoids, saponins, steroids, terpenoids and
tannins. Following standard procedures were used Alkaloids
Determination
The presence of alkaloid was determined using the Mayer and
Dragendorff tests as described by [9 and 10]. 0.2 g of each
extract of the sample were put into a conical flask, 20 ml of
dilute sulphuric acid in methanol were added and then heated
in water bath to boil for 5 min. The mixture was filtered
(vacuum pump) and the filtrates were separated and treated
with 2 drops of Mayer`s (Potassium mercuric iodide solution)
and Dragendorffs (Potassium bismuth iodide solution)
reagents in test-tubes. Development of creamy and an orange
color respectively indicated positive result.

Flavonoids Determination
i. Ammonium Test: The presence of flavonoids in the samples
was determined using the Harbone and Sofowora methods [9
and 11]. 10 ml of ethyl acetate was added to 0.2 g of the
extract and heated in a water bath for 5 min. The mixture was
cooled filtered and the filtrates used for the test.
About 4 ml of filtrate was shaken with 1 ml of dilute ammonia
solution. The layers were allowed to separate and the yellow
color in the ammonical layer (bottom layer) indicates the
presence of flavonoids.
ii. Shinoda Test: Small pieces of Magnesium ribbon followed
by few drops of concentrated hydrochloric acid were added to
a small amount of methanolic extract of the plant material.
Immediate development of a pink scarlet or crimson red color
was taken as an indication of the presence of flavonoids [9 and
10].
Saponins Determination
The froth test and emulsion test as described by Harbone 2001
were used to determine the presence of saponins. 20 ml of
water was added to 0.25 g of the extract in 100 ml beaker and
boiled, filtered and then the filtrates used for the tests:
i. Froth Test: 5 ml of the filtrate was diluted with 20 ml of
water and shaken vigorously. A stable froth (foam) up on
standing indicates the presence of saponins [11]
ii. Emulsion Test: 2 drops of olive oil was added to the
frothing solution and shaken vigorously the formation of
emulsion indicates the presences of saponins.
Steroids / Terpenoids Determination
i. Liebermann-Burchardt test: 1 ml of methanolic extract of
each sample is boiled with 23 ml of acetic anhydride, and
then cooled; 1 to 2 drops of concentrated sulfuric acid were
added slowly through the wall of the tube. Dark green
coloration of the solution indicates the presence of Steroids
and dark pink or red coloration in the interface indicate the
presence of Terpenoids.
ii. Salkowski test: 1 ml of methanolic extract of each sample is
boiled with 2 ml chloroform, cooled, 1 to 2 drops of
concentrated sulfuric acid were added slowly through the wall
of the tube. Shake well and allow standing for some time, red
color appears at the lower layer indicates the presence of
Steroids and formation of yellow colored lower layer indicates
the presence of triterpenoids.
Tannins Determination
i. Ferric chloride test: About 0.5 g of the methanolic extract
was dissolved in 5 to 10 ml of distilled water and filtered. A
few drops of a 10% ferric chloride solution were added to the
filtrate. A greenish black colour or a precipitate was taken as
an indication of the presence of tannins [9 and 10].
ii. Alkaline reagent test: Test solution with sodium hydroxide
solution gives yellow to red precipitate within short time.

79

Journal of Applied and Industrial Sciences, 2013, 1 (3): 78-88, ISSN: 2328-4595 (PRINT), ISSN: 2328-4609 (ONLINE)

Characterization of Agarwood Methanolic Extracts by

Fourier Transforms Infrared (FTIR)
All extracts were characterized by FTIR spectroscopy (Perkin
Elmer 2000 Model) for identification of the active functional
groups. The FTIR study was carried out by using the Perkin
Elmer System 2000 FTIR instrument. First, the transparent
Pellets (thin disc) were formed by mixing 5mg of the sample
with 100 mg of potassium bromide (KBr) (1:20) using a mould
and press, and compressed under a pressure of 7 ton. The
investigation was performed within the wavelength ranging
from 4000 to 400 cm-1 and the spectrum takes about three
minutes to be recorded. The acquisition of the spectra and
peaks assignment was performed using FTIR software
Spectrum Version 3.02.01 (Perkin Elmer, Inc., Waltham,
MA).
Gas chromatography-Mass Spectrometry (GC-MS) Analysis
Gas ChromatographyMass Spectrometry (GCMS) analysis
was carried out for the all methanolic extracts using Perkin
Elmer, Clarus 600 gas chromatograph combined to Perkin
Elmer, Clarus 600 T Mass Spectrometer. The methanolic
extracts were redissolved in methanol and filtered using 0.45
m pore size whatman polyethersulfone membrane filter. The
analysis was performed according to the method described by
Soetardjo et al, 2007. Semi-standard non polar fused silica
DB-1 capillary column (30 m long * 320 m internal diameter)
served as stationary phase. The column temperature was
programmed to 50C for 6 min, with 10C increase per min to
250C, which was maintained for 30 min. The injector and
detector temperatures were both maintained at 250C. Helium
was used as a carrier gas at a flow rate of 1 ml/min; splitting
ratio was: 10:1. The Mass spectral ionization temperature was
set at 250 C. The mass spectrometer was operated in the
electron impact ionization mode at a voltage of 70 eV. Mass
spectra were taken over the m/z range 28400 atomic mass
unit (amu). The compounds of each plant extracts were
identified by computer searches in commercial libraries of
WILEY and NIST (National Institute of Standards and
Technology).
III. RESULTS AND DISCUSSION
The percentage yield of the extracts was based on the weight
of dried and ground plant materials. Beside the single solvent
extraction, four solvents extraction was used in another study,
namely n-hexane (C6H14), dichloromethane (CH2Cl2), ethyl
acetate (EtOAc) and methanol (CH3OH). The percentage yield
of the methanolic extracts of Agarwood leaves (MOB and
MOA) were shown in Table 1.
Table 1
The physical properties of dry crude methanolic extracts of the
Agarwood leaves
Extract
Physical characteristics
Percentage yield (%)
Color
Odor
Consistency

MOB extract

MOA extract

52.12
Dark green
Leafy smell but
sweet odor
Waxy thick

39.6
Dark green
Leafy smell but
sweet odor
Waxy thick

Preliminary Phytochemical Analysis of the Agarwood

Methanolic Crude Extracts
The qualitative screening of phytochemical compounds in
Agarwod methanolic extracts revealed the presence of
alkaloids, saponins and phenolic compounds (flavonoids,
terpenoids, and tannins). The presence of phytochemicals
indicates its potential as a source of useful corrosion inhibitors.
Factors affecting capture of active phytochemicals are the
plant part being extracted, the types of solvents, the extraction
period and extraction conditions [12 and 13]. For example,
antioxidant in plant material may be water soluble, fat soluble,
insoluble or bound to cell walls, and react at different rates.
Therefore, methanol was chosen in this study as the organic
solvent for its wide solubility properties for low molecular
weight and moderately polar substances, including phenolic
compounds. However, despite these challenges, qualitative
screening of native plants is useful for revealing activity that
may lead to the development of new products for use as
anticorrosion.
The phytochemical compounds of plants have potentially
significant application in human health care as well as in
corrosion field. The presence of each secondary metabolite in
Agarwood leaves extracts gives a justification for the
traditional use of the plant in treating various health problems.
For instance, detection of alkaloids in Agarwood leaves
extracts was of great importance, since significant quantities of
alkaloids could be used as antimalaria, analgeics,
antispasmodic, bactericidal and stimulants, which are all
pharmaceutical properties of the plant. Presence of alkaloids in
Agarwood leaves extracts, justifies the use of the plant to treat
toothache, colic, severe headache, rheumatism and pains
during pregnancy.
On the other hand, flavonoids, which are a group of
polyphenolic compounds, have been reported to have antiinflammatory action, free radical scavenging and inhibition of
hydrolytic and oxidative enzymes [14], anti-allergic, anticancer, anti-inflammatory and anti-viral [15]. Moreover, the
presence of glycosides moieties like saponins, anthraquinones,
cardiac glycosides and flavonoids could inhibit tumor growth
and protect against gastrointestinal infections [16 and 17].
Corrosion inhibition performance of three flavonoids,
apigenin, luteolin-3-methyl ether and quercetin-3,3dimethylether on copper dissolution in 2.0 M HNO3 was
investigated by [18]. 92% Inhibition was observed in some of
these flavonoids.
The inhibitive action of commercial green tea extracts flavonoids monomers of tea catechins on mild steel (MS) in a
1.0 M hydrochloric acid solution was investigated by [19],
84% Inhibition was observed in one of the green tea extracts.
Herbs that have tannins as their components are astringent in
nature, i.e., fasten the healing of wounds and inflamed mucous
membranes and could be used for treating intestinal disorders
such as diarrhea and dysentery exhibiting antibacterial activity
[20]. Furthermore, polyphenols are known to be responsible
for antioxidant and antimicrobial activities [20 and 21]. Recent

80

Journal of Applied and Industrial Sciences, 2013, 1 (3): 78-88, ISSN: 2328-4595 (PRINT), ISSN: 2328-4609 (ONLINE)
epidemiological studies have strongly suggested that
consumption of certain plant materials rich in phenolic
compounds may reduce the risk of chronic diseases related to
oxidative stress on account of their antioxidant activity and
thus promote general health benefits. Therefore, the significant
antibacterial, anticandida and antioxidant activities of
Agarwood plant could be related to the presence of phenolic
compounds.
Catechin,
epigallocatechin
gallate,
epicatechingallate,
epigallocatechin and epicatechin that constitute Mangrove
tannins were investigated in an aerated HCl, the monomers
were found to be mainly cathodic inhibitors and the inhibition
efficiency was dependent on concentration.
Saponin has the property of precipitating and coagulating red
blood cells. Some of the characteristics of saponins include
formation of foams in aqueous solutions, hemolytic activity,
cholesterol binding properties and bitterness [22]. These

properties bestow high medicinal activities on the extracts. It

has also been shown that saponins are active antifungal agents
[23]. This therefore supports the earlier finding that extracts of
the plant used in the present work may be useful in the
chemotherapy of mycotic infections.
Presence of tannins suggests the ability of this plant to play a
major role as antidiarrhoec and antihaemorrhagic agent [24].
The presence of mimosa tannin as a low carbon steel corrosion
inhibitor in sulphuric acid media was tested [25] in
concentrations from 105 to 101 mol/l, at the temperature of
298 K in the solutions of pH 1, 2 and 3. The inhibitor
effectiveness increases with increase in concentration.
Beside the anti corrosion activity of methanol soluble leaf
extracts, the present study indicates the usefulness of
agarwood tree for medicinal purposes. The presence of
phytochemicals indicates its potential source of useful drugs
(further study is recommended).

Table 2
Summary of the phytochemical screening of crude Methanolic Extracts of Agarwood leaves (before and after inoculation)
Extract

Before Inoculation
(MOB)

Test

Observation

Alkaloids test (Mayers

Reagent)

Creamy ppt.

Alkaloids test
(Dragndorffs Reagent)

After Inoculation
(MOA)
Result

Observation

Result

Dense Creamy ppt.

Orange red ppt.

Reddish brown ppt

Triterpenoid / Steroid
(Salkowski-Liberman)
Flavonoids test
(Harbone and Sofowora
methods)
-Ammonium test
Saponin test
(Froth test)

Reddish brown ring in

the interface

+T

Yellow color obtained

in the ammonical layer
(lower layer)

Stable persistent
froth(thick layer)

Saponin test-Emulsion
test

Formation of emulsion

Formation of
emulsion

Greenish black
solution (ppt.)

Dark green solution

Tannin test (Harbone,
(ppt.)
Braemers methods)
+: Positive result (Presence of the phytochemical)
+T: Positive result for triterpenoid

Reddish brown ring

in the interface
Clear yellow color
obtained in the
ammonical layer
(lower layer)
Stable persistent
froth(thin layer)

+T

81

Journal of Applied and Industrial Sciences, 2013, 1 (3): 78-88, ISSN: 2328-4595 (PRINT), ISSN: 2328-4609 (ONLINE)
Table 3
Summary of the phytochemical screening tests on Methanolic crude extracts of Agarwood leaves (Before and After inoculation)

Crude
Extract

Alkaloids
test
(Mayers
Reagent)

Alkaloids test
(Dragndorffs
Reagent)

Flavonoids
test
(Harbone
and
Sofowora
methods)

Triterpenoid /
Steroid
(SalkowskiLiberman)

Saponin
test
(Froth
test)

Saponin test
(Emulsion
test)

Tannin test
(Harbone,
Braemers
methods)

+T

++

++

+++

++

++

++T

++

MOB

MOA

+: Positive result: + Low, ++ Medium, +++: High

+T: Positive result for Triterpenoid Medium, ++T: High
Characterization by Fourier Transform Infrared (FTIR)
analysis
The infrared spectra of all the extracts were taken and
compared. The spectrums did not shows significant changes of
peaks for Agarwood leaves before and after inoculation
process. Figures 1a, 1b show the IR spectrum of MOB, MOA
respectively. Figure 2 shows the infrared spectra for all the
methanolic extracts of Agarwood leaves before and after
inoculation process with its adsorption characteristic region for
OH, C=C and C-O groups (in horizontal line). The spectrums
did not show significant changes of peaks for Agarwood
leaves before and after inoculation process.
FTIR spectrum of methanol only extract before inoculation
(MOB) shows the main characteristic absorption broad band
appeared at 3388 cm-1 assigned to the presence of (OH/NH)
group; the presence of the methyl (CH3) and methylene (
CH2) aliphatic saturated (CH) sharp asymmetric and
symmetric stretching band which are observed at 2924 cm-1
and 2852 cm-1, respectively. The (C=O) stretching band is
observed at 1709 cm-1; it is due to the presence of carboxylic
acid/ketones groups. The band at 1621 cm-1 shows the
presence of aromatic ring (C=C).The peak at 1455 cm-1 is due
to aromatic (CC) stretching band. The peak at 1280 cm-1 is
due to aromatic (COC) band. The (CN) medium stretching
band and/or PH phosphine band were observed at 1076 cm-1.

Absorption band appeared at 827 cm-1 assigned to (SOR)

esters. Furthermore, the adsorption band around ~ 1200900
cm-1 and~ 1100700 cm-1 may represents to the 1,2disubstituted and 1,3-disubstituted benzene ring [26], (Figure
1a).

FTIR spectrum of methanol only extract after inoculation

(MOA) shows (Figure 1b) the main characteristic absorption
broad band appeared at 3384 cm-1 assigned to the presence of
(OH/NH) group; the presence of the methyl (CH3) and
methylene (CH2) aliphatic saturated (CH) sharp asymmetric
and symmetric stretching ba nd which are observed at 2923
cm-1 and 2851 cm-1, respectively. The (C=O) stretching band is
observed at 1708 cm-1; it is due to the presence of carboxylic
acid group. The band at 1618 cm-1 shows the presence of
aromatic ring (C=C).The peak at 1463 cm-1 is due to aromatic
(CC) stretching band. The peak at 1290 cm-1 is due to
aromatic (COC) band. The (CN) medium stretching band
and/or PH phosphine band were observed at 1078 cm-1. The
aromatic bending band occurs at the range of 827-787 cm-1.
Absorption band appeared at 827 cm-1 assigned to (SOR)
esters. Furthermore, the adsorption band around ~ 1200900
cm-1 and~ 1100700 cm-1 may represents to the 1,2disubstituted and 1,3-disubstituted benzene ring [26], (Figure
1b).

82

Journal of Applied and Industrial Sciences, 2013, 1 (3): 78-88, ISSN: 2328-4595 (PRINT), ISSN: 2328-4609 (ONLINE)
66.4
65

60

55
827
787

50

611

%T
45
1167

40

1377
1455

1280

2852
1076

35
1621
3388

2924

29.4
4000.0

3000

2000

1500

1000

500 370.0

cm-1

Fig. 1a: FTIR Spectrum for Agarwood methanolic extract before inoculation (MOB)

76.4

74

72

70

610

%T 68
66
1730

64

1708
2851

1290
14631380

3384

62

2923

1078

1618

60.1
4000.0

3000

2000

1500

1000

400.0

cm-1

Fig. 1b: FTIR Spectrum for Agarwood methanolic extract after inoculation (MOA)
The presence of atheroline alkaloids was supported by using
FTIR studies. It showed the presence of aromatic rings (C=C
stretching bands) between 1621 and 1624 cm-1 and the O-Me
group appeared from 1383 to 1075 cm-1. Moreover, a broad
band of the FTIR spectrum at around 3400 cm-1 showed the
presence of N-H groups.
Gas Chromatography-Mass Spectrometry (GC-MS) Analysis
Fourteen different compounds were identified by GC-MS
analysis of the Agarwood leaves methanolic extracts.
Hexadecanoic acid was found to be one of the major
compounds in Agarwood leaves methanolic extracts. It was
eluted at the same retention time with large abundance.

Hexadecanoic acid was one of the most common saturated

fatty acids found in animals and plants, and was reported to
have potential antibacterial and anti fungal activity. In one
study by Saidana, [27], Hexadecanoic acid had been reported
to exhibit antibacterial activity against Gram-positive and
Gram-negative bacteria comparable to ampicillin and in the
case of Streptococci, greater than that of ampicillin. In another
study by [28], Hexadecanoic acid was considered as the major
antibacterial compound in the ethyl acetate root extract of
Pentanisia prunelloides. Thus, the presence of Hexadecanoic
acid could justify the use of Agarwood leaves methanolic
extracts as antibacterial and anti fungal as well as anti
corrosion especially in that type of corrosion caused by

83

Journal of Applied and Industrial Sciences, 2013, 1 (3): 78-88, ISSN: 2328-4595 (PRINT), ISSN: 2328-4609 (ONLINE)
bacteria in underground pipes. So further study of
Hexadecanoic testing as anti corrosion in microbiological
corrosion is recommended.
The gas chromatograms of the methanolic extracts of A.
Malaccensis leaves are shown in Figures 3a and 3b while the
compounds and their corresponding mass spectra are shown in
Table 5 and 6, the retention times (Rt) were reported in
minutes. The major compounds identified in MOB extract

were 1, 4, 7, 10, 13-Pentaoxacyclopentadecane (Rt 2.52,

13.9%); 1, 4, 7, 10, 13, 16-Hexaaoxacyclooctadecane (Rt 4.44,
8.17%), Acetic acid (Rt 6.2, 6.87%) and Hexadecanoic acid
(Rt 19.82, 4.15%); in MOA were 1, 4, 7, 10, 13Pentaoxacyclopentadecane (Rt 2.52, 20.6%), Hexaethylene
glycol monododecyl ether (Rt 3.8, 21.56%) and 2, 6, 10, 14,
18, 22- tetracosahexaen, 2, 6, 10, 15, 19, 23-hexamethyl- (Rt
21.17, 4.43%);

Table4
The summary of characteristic peaks bands on FTIR spectra for all the methanolic extracts
Wavenumber (cm-1)
Functional groups
(bands)
MOB extract
MOA extract
3388
3384
OH (stretch)
(b),(str)
(b),(str)
CH alkane stretching
2924(str)
2923(str)
C=O stretching
1709(m)
1708(str)
C=C aromatic
1621
1618
stretching
(b), (str)
(b), (str)
Asymmetric
1511(str)
1509(str)
RCOO- stretching
CC stretching
1455(str)
1463(str)
Aromatic hydroxyl
1377(str)
1380(str)
deformation
COC cyclic ether
1280(str)
1290(str)
Aromatic CO
1167(m)
1166(str)
stretching
Aliphatic CO/CN/P1076(str)
1078(str)
H stretching
(Note; (b): broad, (s): strong, (m): medium, (w): weak)
C=C

C-O

OH

Fig2: Fourier transform infrared spectrum. The black spectrum) methanolic extract corrosion inhibitor (MOB) (uninoculated
leaves); and the blue spectrum) methanolic extract corrosion inhibitor (MOA) (inoculated leaves);

84

Journal of Applied and Industrial Sciences, 2013, 1 (3): 78-88, ISSN: 2328-4595 (PRINT), ISSN: 2328-4609 (ONLINE)
Gas chromatography-mass spectrometry (GC-MS) analysis of
plant extracts led to the identification of 14 components from
the two types of Agarwood leaves extracts (Table 7). It is
interesting to see here that all identified compounds from plant
extracts contained oxygen and/or -electrons in their
molecules. Moreover, from the previous studies on the
phytochemical constituents of the plant extracts it was
established that the plant extracts used in this study also
contain a mixture of organic compounds containing O, N or electrons in their molecules. Hence, the corrosion inhibition of
carbon steel through these studied plants may be attributed to
the adsorption of the phytochemicals containing O, N or -

electrons in their molecules as these atoms are regarded as

centers of adsorption onto the metal surface. However, the
highly complex chemical compositions of the plant extracts
make it rather quite difficult to assign the inhibitive effect to a
particular compound present in plants extracts. Having
confirmed the corrosion inhibition effectiveness of these plants
extracts, further detailed investigation for each plant extracts
through inhibitive assay guided isolation using surface
analytical techniques will enable the characterization of the
active compounds in the adsorbed layer and assist in
identifying the most active phytochemicals.

Fig. 3a: Gas chromatogram of the Methanolic extracts of Agarwood leaves before inoculation (MOB)

Fig. 3b: Gas chromatogram of the Methanolic extracts of Agarwood leaves after inoculation (MOA)

85

Journal of Applied and Industrial Sciences, 2013, 1 (3): 78-88, ISSN: 2328-4595 (PRINT), ISSN: 2328-4609 (ONLINE)
Table5:
Results of the GC-MS analysis of Agarwood leaf extract (MOB)
Percent
No.

Rt(min)

of total

Compound

Structure

(%)

19.816

4.153

23.207

3.382

n-Hexadecanoic
acid, methyl ester

9,12,15octadecatrienoic
acid,(Z,Z,Z)-

2,3,Dihydro-3,53

14.632

Dihydroxy-6-

methyl-4H-pyran4-one

Table6:
Results of the GC-MS analysis of Agarwood leaf extract (MOA)
Percent
No.

Rt(min)

of total

Compound

(%)

n-Hexadecanoic
1

19.816

3.876

acid, methyl
ester

17.672

0.629

14.992

0.921

Phytol

1,2,3propanetriol

Structure

86

Journal of Applied and Industrial Sciences, 2013, 1 (3): 78-88, ISSN: 2328-4595 (PRINT), ISSN: 2328-4609 (ONLINE)
Table 7:
Chemical components identified in Agarwood leaves methanolic extracts by GC-MS.
Molecular
No. Component
MOB
MOA
Formula
C5H10O4

C12H16O6

n-hexadecanoic acid

C16H32O2

Glycerine

C3H8O3

Squalene

C12H24O6

2-propanone,1,3 dihydroxy

C3H6O3

1,2,3-propanetriol, monoacetate

Phenyl--D-glucoside

3,7,11,15-Tetramethyl-2-Hexadecen7

C20H40O

1-ol (Phytol)
8

Dodecyl acrylate

C15H28O2

C6H8O4

C15H16O6

2,3-Dihydro-3,5-Dihydroxy-6-methyl9
4H-pyran-4-one
6-ethyl-5-hydroxy-2,3n,710
trimethoxynaphthoquinone

11

9,12,15-Octadecatrienoic acid, (z,z,z)-

C18H30O2

12

Squalene

C30H50

13

Octaethylene glycol monodecyl ether

C28H58O9

14

1-Tetradecanol

C14H30O

(+): present

(): absence

ACKNOWLEDGEMENT
Greatest thanks to Dr. Afidah Abdul Rahim- Universiti Sains
Malaysia (USM), School of Chemical Sciences, PenangMalaysia for providing the leaves of Aquilaria Malaccensis

and for giving a great opportunity to use USM - School of

Chemical Sciences` corrosion labs.

87

Journal of Applied and Industrial Sciences, 2013, 1 (3): 78-88, ISSN: 2328-4595 (PRINT), ISSN: 2328-4609 (ONLINE)

REFERENCES
[1] Satapathy, A. K., Gunasekaran, G., Sahoo, S. C., Kumar Amit,
Rodriques, P. V. (2009). Corrosion inhibition by Justicia gendarussa
plant extract in hydrochloric acid solution. Corrosion Science 51,
2848-2856.
[2] Gunasekera S. P, Kinghorn A. D, Cordell G. A, Farnsworth N. R
(1981). Plant anticancer agents, XIX. Constituents of Aquilaria
Malaccensis. J. Nat. Prod. 44: 569-572.
[3] Okugawa, H.; Ueda R, Matsumoto, K.; Kawanishi K, Kato A
(1993). Effects of agarwood extracts on the central nervous systems
in mice. Planta Med. 59:32-36.
[4] Pranakhon, R.; Pannangpetch, P. and Aromdee C., 2011.
Antihyperglycemic activity of agarwood leaf extracts in STZinduced diabetic rats and glucose uptake enhancement activity in rat
adipocytes. Songklanakarin J. Sci. Technol. 406 33 (4), 405-410.
[5] Mabberley, D. J. (1997). The Plant Book. The Press Syndicate of
the University of Cambridge, UK. 858pp.
[6] Chakrabarty, K., Kumar, A. and Menon, V. (1994). Trade in
Agarwood. TRAFFIC India and WWF-India, New Delhi. 51pp.
[7] Wiriadinata, H. (1995). Gaharu (Aquilariaspp.) Pengembangan
dan Pemanfaatan yang Berkelanjutan. In Lokakarya Pengusahaan
Hasil Hutan Non Kayu (Rotan, Gaharu, dan Tanaman Obat).
Departement Kehutanan. Indonesia-UK Tropical Forest Management
Programme. Surabaya.
[8] Oldfield, S., Lusty C., and MacKinven A. (1998). The Word List
of Threatened Trees. World Conservation Press, Cambridge, UK.
650pp.
[9] Harbone, J. B. (2001). Phytochernical methods. Chapman and
Hall Ltd: London, pp111-113.
[10] Ghani, A. (1998) Medicinal plants of Bangladesh, pp. 7883.
(Dhaka: Asiatic Society of Bangladesh).
[11] Sofowora A., (2005). Medicinal plants and traditional medicine
in Africa. Spectrum Books Ltd: Ibadan. Nigeria, p. 289.
[12]Sreelatha, S. and Padma, P.R. (2009). Antixidant activity and
total phenolic content of Moringa oleifera leaves in two stages of
maturity. Plant Foods for Human Nutrition, 64, 303-311.
[13] Borchardt, J. R., Wyse, D.L., Sheaffer, C.C., and Kauppi, K.L.
(2008). Antioxidant and antimicrobial activity of seed from plants of
the Mississippi river basin. Journal of medicinal Plants Research,
2(4), 081-093.
[14] Sannigrahi, S., Mazuder, U.K., Parida., S., and Jain, S. (2010).
Antioxidant potential of crude extract and different fractions of
Enhydra Fluctuans Lour. Iranian Journal of Pharmaceutical Research,
9(1). 75-78.

[15] Agrawal, P. K. Carbon-13NMR of flavonoids. Elsevier science

publishers, Amsterdam, 1989.
[16] Akinpelu, D. A. and Onakoya, T.M. 2006. Antimicrobial
activities of medicinal plants used in folklore remedies in southwestern. African Journal of Biotechnology. 5 (11), pp. 1078-1081.
[17] Abubakar, E. M., (2009). Antibacterial activity of crude extracts
of Euphorbia hirta against some bacteria associated with enteric
infections. Journal of medicinal Plants Research, 3(7), 498-505.
[18] Al-Qudah, M. A. (2010). Inhibition of Copper Corrosion by
Flavonoids in Nitric Acid. E-Journal of Chemistry, 8(1), pp 326-332.
[19] Nofrizal, S.; Afidah A. Rahim, Bahruddin Saad, R. Bothi,
Affaizza M. Shah, and Yahya S., (2011). Elucidation of the
Corrosion Inhibition of Mild Steel in 1.0 M HCl by Catechin
Monomers from Commercial Green Tea Extracts. Metallurgical and
Materials Transactions A. 12 p.
[20] Rasool, R., B.A. Ganai, S. Akbar, A.N. Kamili and A. Masood,
2010. Phytochemical screening of Prunella vulgaris L.: An important
medicinal plant of Kashmir. Pak. J. Pharm. Sci., 23: 399-402.
[21]Zhang, M.; Chen, H.; Li, J.; Pei, Y. & Liang, Y. (2010).
Antioxidant properties of tartary buckwheat extracts as affected by
different thermal processing methods. LWT- Food Science and
Technology, 43, pp.181-185.
[22]Sodipo, O. A., Akiniyi JA, Ogunbamosu J. U (2000). Studies on
certain Characteristics of extracts of bark of pansinystalia macruceras
(K schemp) pierre Exbeille. Glob. J. Pure Applied Science (6), pp
83-87.
[23] Sodipo, O. A, Akanji, M. A., Kolawole F. B., Adutuga O. O.
(1991). Saponin is the active antifungal principle in Garcinia kola,
heckle seed, Bioscience Research Committee (3) p 171.
[24] Asquith, T. N, Butler L. G. (1986). Interaction of condensed
Tannins with selected proteins. Phytochemistry 25 (7) 1591-1593.
[25]Martinez, S. and Stern I. (2002). Applied Surface Science; 199, p
83.
[26] Kim, S., T. (2009). Transformation of Rust by Uncaria gambir,
Master Dissertation, Universiti Sains Malaysia, p 13-46.
[27] Saidana, D., Mahjoub, S., Baoussaada, E., Chariaa J., Mahjoub,
M.A., Cheraif, I., DAAMI, D., Mighri, Z., and Helal, A.N. (2008).
Antibacterial and antifungal activities of the essential oils of two
Saltcedar species from Tunisia. Journal of the American Oil
Chemists. Society, 85(9), 817-826.
[28]Yff, B. T. S., Lindsey, K.L., Taylor, M, B., rasmus, D.G., and
Jager, A, K. (2002). The pharmacological screening of Pentanisia
prumelloides and the isolation of the antibacterial compound palmitic
acid. Journal of Ethnopharmacology, 79(1), 101-107.

88