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RESEARCH ARTICLE
Year : 2009  |  Volume : 1  |  Issue : 5  |  Page : 270-273 Table of Contents     

Antioxidant activity of Aquilaria malaccensis (thymelaeaceae) leaves


1 Department of Chemical Sciences, Faculty of Science & Technology, Universiti Malaysia Terengganu, Malaysia
2 Institute of Marine Biotechnology, Universiti of Malaysia Terengganu, Malaysia
3 Malaysian Nuclear Agency (MNA), Kajang, Selangor, Malaysia

Date of Submission22-Apr-2009
Date of Decision15-May-2009
Date of Acceptance25-May-2009
Date of Web Publication2-Jan-2010

Correspondence Address:
A.W.N Huda
Department of Chemical Sciences, Faculty of Science & Technology, Universiti Malaysia Terengganu
Malaysia
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Source of Support: None, Conflict of Interest: None


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   Abstract 

The phytochemical and antioxidant activity of Aquilaria malaccensis leaves were investigated. The sequential maceration extraction methods utilizing solvents with different polarities namely hexane, ethyl acetate and methanol yielded the corresponding crude extract. The extracts were subjected to preliminary phytochemical screening and revealed the presence of alkaloids, flavanoids, triterpenoids, steroids and saponins. The phytochemical screening suggests that flavanoids present in this species might provide a great value of antioxidant activity. Preliminary screenings of the free radical scavenging activity on the extracts of the plants with 2, 2-Diphenyl-1-picrylhydrazyl (DPPH) were tested and showed positive result. Quarcetine was used as reference standard. The extracts exhibited strong antioxidant activity radical scavenging activity with IC 50 value of 8.0 Χ 10 2 μg/ml, 1.6 Χ 10 2 μg/ml, 1.4 Χ 10 2 μg/ml, 30.0 μg/ml and 3.33 μg/ ml for hexane, DCM, ethyl acetate, methanol and quarcetine respectively.

Keywords: Aquilaria malaccensis , IC 50 value, antioxidant activity


How to cite this article:
Huda A, Munira M, Fitrya S D, Salmah M. Antioxidant activity of Aquilaria malaccensis (thymelaeaceae) leaves. Phcog Res 2009;1:270-3

How to cite this URL:
Huda A, Munira M, Fitrya S D, Salmah M. Antioxidant activity of Aquilaria malaccensis (thymelaeaceae) leaves. Phcog Res [serial online] 2009 [cited 2017 Mar 29];1:270-3. Available from: http://www.phcogres.com/text.asp?2009/1/5/270/58053


   Introduction Top


Antioxidants play a major role in helping to protect our body from the formation of free radicals and prevent or delay the occurrence of lipid peroxidation. In our body, oxidants and free radicals, which are formed from triplet oxygen, water and unsaturated lipid molecules, can caused oxidative stress in tissue of lungs, heart and cardiovascular system, kidneys, liver, gastrointestinal tract, blood, eye, skin, muscle and brain. Thus, free radical is to be major precursor for the development of various degenerative diseases, such as cancer, atheroschlerosis, gastric ulcer, diabetic, diabetic and others. An extensive study has been carried out to evaluate antioxidant property of compounds originated from terrestrial plant sources (Habsah et al., 2006). Phenolics plant, in particular phenolic acids, tannins and flavanoids are known to be as potent antioxidants (Pratt & Hudson, 1990). In this context, one such plant is Aquilaria Malaccensis. Aquilaria Malaccensis commonly known as gaharu is an aromatic plants.

Aquilaria malaccensis is widely distributed in south and south-east Asia. There are differing accounts of the countries in which it occurs. According to Oldfield et al. (1998), A. malaccensis is found in 10 countries which are Bangladesh, Bhutan, India, Indonesia, Iran, Malaysia, Myanmar, Philippines, Singapore and Thailand. Aquilaria malaccensis is prescribed in traditional East Asian medicine to relieve pain, arrest vomiting by warming the stomach, and to relieve asthma (Anon, 2003). High-grade agarwood powder is prescribed in Chinese medicine and is also used in the production of pharmaceutical tinctures (Van Beek & Phillips, 1999). The phytoconstituents isolated so far from this leaves are kusunol, jinkoh-eremol, jinkohol II, α-agarofuran, (-)-10-epi-γ-eudesmol and oxo-agarospirol, 10-epi-γ-eudesmol (Yoneda et al. 1984) and six new 2-(2-Phenylethyl) chromone compounds (Konishi et al. 2002). According to Karim et al. (2000), A. malaccensis leaves contained sesquiterpene alcohols, which was 10-epi-γ-eudesmol. There are few studies have been done previously on phytochemical screening and antioxidant activity using A. malaccensis extracts. Therefore, this study was focused on the antioxidant activity of leaves extract. Different approached had been use to enhance the research.


   Chemicals Top


2, 2-diphenyl-1-picrylhydrazyl (DPPH) and quercetin were obtained from Organic Laboratory, Department of Chemical Sciences. All the chemicals and reagents used were of analytical grade.


   Plant Materials Top


The fresh leaves of Aquilaria malaccensis were collected from Malaysian Nuclear Agency (MNA), Kajang, Selangor. The sample was then identified by one of the research officer of Malaysian Nuclear Agency.


   Extraction Top


Air dried leaves (977 g) was ground into fine powder and extracted by continuously soaking with hexane, followed by dichloromethane, ethyl acetate and methanol solvent. The crude extracts were concentrated under reduce pressure to give solid residue using rotary evaporator. The yield was found to be 1.78, 2.34, 5.26 and 7.28% w/w with reference to the air dried plant.


   Preliminary Phytochemical Screening Top


Phytochemical screening test was a qualitative method which was conducted to ensure the presence of alkaloids, steroids, triterpenoids and flavonoids in each crude extract. There were four main test which are Alkaloid test (Mayer), Triterpenoid / Steroid test (Liebermann-Burchard), Saponin test and Flavonoid test. The crude of hexane, dichloromethane, ethyl acetate and methanol was subjected into four tests.


   Antioxidant Activity Top


The DPPH free radical scavenging assay was conducted on the hexane, dichloromethane, ethyl acetate and methanol crude extracts using modified method. The crude extracts (10 mg) were dissolved in 10 μl dimethylsulfoxide (DMSO). Then a clear microtiter plate was added with 5 μl DMSO solution and 5 μl of samples were added to the plate. The following concentrations of extract were prepared that is, 10, 5, 2.5, 1.25, 0.625, 0.3125 and 0.15625 mg/ml. Quarcetine was used as standard or positive control. All the solutions were prepared in the microtiter plate with DMSO. DMSO was used as the blank sample of this experiment. The experiments were carried out in replicate. The plate was left to incubate at room temperature for 30 minutes in the dark. The reduction of the DPPH free radical was measured by reading the UV absorbance at 517 nm by ELISA instrument. All tests and analyses were run in triplicate and averaged.

Percentage inhibition was calculated using the following formula:



Abs: absorbance

To obtain the IC 50 value, the stocks of Quercetin (1 mg/ml) and crude extract (10 mg/ml) were diluted (two fold dilution) in 96-well micro plates to varying concentration topping from 1000 μg/ml down to the lowest of 15.6 μg/ml. The free radical-scavenging activities were determined as above (Habsah et al., 2006). A dose response curve was plotted to determine the IC50 values. IC 50 is defined as the concentration sufficient to obtain 50% of a maximum scavenging capacity.


   Results and Discussion Top


Preliminary phytochemical screening

Preliminary phytochemiacal screening in all crude showed the presence of alkaloids, steroids, tritepenoids, saponin and flavonoids. All crude except for ethyl acetate crude contains flavonoids which are generally potent inhibitors of free radicals (Havsteen, 1983). [Table 1] showed the obtained result from phytochemical screening.

Antioxidant Activity-DPPH assay

This screening was done to determine whether there are any antioxidant properties in the crude. [Figure 1] shows the percentage of inhibition H-donor activity of quercetin as measured using DPPH assay with different concentration and [Figure 2] shows the percentage of inhibition H-donor activity of hexane, dichloromethane, ethyl acetate and methanol crude extract as measured using DPPH assay with different concentration.

DPPH assay clearly shows the highest potential of antioxidant properties in each crude extracts of A. malaccensis especially the methanol crude extract. [Table 2] shows the percentage of DPPH Free Radical Scavenging Activity of each crude extracts while [Table 3] shows the percentage of DPPH Free Radical Scavenging Activity of quarcetine as the standard used in this study.

The IC 50 value of each crude extracts is the inhibition concentration to obtain 50% of a maximum scavenging capacity. The DPPH free radical scavenging activity for each crude extract can be simplifies as shown in [Table 4], respectively. It could be observed that all the crude extracts exhibited a positive DPPH free radical scavenging activities. The IC 50 values of DPPH free radical scavenging activity was in decreasing order:

MeOH > DCM > EA > Hexane

The methanol crude possessed the highest antioxidant activity than DCM, EA and Hexane crude extract. Methanol extracts were the most effective DPPH scavengers. Quarcetine is a potent free radical scavenging. So when compared to such pure compounds, IC 50 value of the different crude extract is quite good proving that they are potent DPPH free radical scavenger. This can be attributed to the presence of flavonoids contains in the extract. Therefore, the extended researches are needed for the isolation and identification of the antioxidant compounds.


   Conclusion Top


In conclusion, the preliminary phytochemical screening of the fruits of A. malaccensis indicates the presence of secondary metabolities, having an essential role in medicine. Overall, this study indicates the antioxidant activity of A. malaccensis and provides some idea about phytochemical investigation on A. malaccensis.


   Acknowledgment Top


The authors wish to thank Faculty of Science & Technology, Institute Marine Biotechnology, Universiti Malaysia Terengganu and Malaysian Nuclear Agency (MNA), for laboratory and equipments provided. [9]

 
   References Top

1.Anon, 2003, Review of Significant Trade Aquilaria malaccensis Barden A., Awang A. N, Mulliken T. and Song M., Heart of The Matter: Agarwood Use and Trade and Cities Implementation For Aquilaria malaccensis  Back to cited text no. 1      
2.Mohamad Habsah, Nur Najmiah Wan Abdul Jamil Wan, Abas Faridah, Alwi Afnani, Suryati Mohamad Khamsah, Fitrya Syamsumir Desy & Ch-ing Yee Tee. 2006. Antioxidant activity of crude extracts from selected ma-rine sponges collected off Terengganu Islands. 5 th ANRAP international Seminar-MNPS 22 nd Annual seminar 2006, 8-10 November 2006, Kuala Lumpur, Malaysia.  Back to cited text no. 2      
3.Havsteen B. Flavanoids, a class of natural products of high pharmacologi-cal potency. Biochem Pharmacol. Vol. 32 pp, 1141-1148 (1983).   Back to cited text no. 3      
4.Karim Y., Joulain D., Robertet S.A., Volatile Constituents from The In-fected Wood of Aquilaria malaccensis Benth, Sains Malaysiana, Vol. 29, pp. 197-200, Grasse, France (2000).  Back to cited text no. 4      
5.Konishi T., Konoshima T., Shimeda Y., Kiyosawa S., Six New 2-(2-Phenyleth-yl)chromones from Agarwood, Vol. 50, No. 3, pp. 419-422 (2002).  Back to cited text no. 5      
6.Oldfield S., Lusty C., MacKinven A. (1998). The Word List of Threatened Trees. In: Barden, A., Awang Anak Noorainie, Mulliken T., and Song M. (2000). Heart of the matter: Agarwood use and trade and CITES implementation for Aquilaria malaccensis. TRAFFIC International  Back to cited text no. 6      
7.Pratt, D.E & Hudson, B.J.F. (1990). Natural Antioxidants Not exploited com-mercially- Food Antioxidant. Amsterdam: Elsevier.  Back to cited text no. 7      
8.Van Beek H., Phillips D. Agarwood: Trade and CITES Implementation in South-east Asia. Unpublished report prepared for TRAFFIC Southeast Asia, Ma-laysia (1999).  Back to cited text no. 8      
9.Yoneda K., Nakanishi T., Yamagata E., Nagashima T., Kawasaki I., Yoshi-da T., Mori H., Miura I. Three Fragrant Sesquiterepenes of Agarwood, Phytochemistry, Vol 23, No. 11, pp. 2066-2067 (1984).  Back to cited text no. 9      


    Figures

  [Figure 1], [Figure 2]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4]



 

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  In this article
    Abstract
    Introduction
    Chemicals
    Plant Materials
    Extraction
    Preliminary Phyt...
    Antioxidant Activity
    Results and Disc...
    Conclusion
    Acknowledgment
    References
    Article Figures
    Article Tables

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