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PHYTOCEMICAL ANALYSIS AND ANTI OKSIDANT ACTIVITY OF THE WOOD ETHANOLIC EXTRACTS OF SIRIH HUTAN (Pipe aduncum)
Erwin1
Department of Chemistry, Faculty of Mathematic and Natural Sciences, Mulawarman University, Jalan Barong Tongkok 4, Samarinda 75119, Indonesia
Abstrak. Sirih hutan (Piper aduncum) tumbuh secara merata di hutan hujan tropis Kalimantan Timur. Tumbuhan ini digunakan sebagai obat gatal-gatal oleh masyarakat setempat. Penelitian ini bertujuan untuk menentukan kandungan kimia dan mengevaluasi potensi aktivitas antioksidan dari ekstrak kayu hutan Sirih. Berdasarkan hasil GC-MS dari ekstrak kayu Sirih hutan terdiri dari 11 senyawa. Sedangkan daya hambat terhadap peredaman radikal bebas DPPH dari ekstrak etanol adalah IC50 = 259,74 ppm.
Kata kunci: Sirih hutan, aktivitas antioksidan, dan GC-MS
Abstract. Sirih hutan (Piper aduncum) grew evenly in the tropical rain forests of East Kalimantan; it was used as a drug itching by local people. The present study is aimed to determine the chemical content and evaluate the free radical scavenging potential antioxidant activity of the extract of Sirih hutan wood. Based on the results of the GC-MS of the wood extract of Sirih hutan consists of 11 compounds. The inhibition of DPPH scavenging free radicalst of extract in ethanol preparations is IC50 259.74 ppm.
Keyword: Sirih hutan, antioxidant activity, and GC-MS
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INTRODUCTION
Piperaceae family consists of about 40 genera and piper is one of the main genus comprising about 700 species distributed throughout the tropical and subtropical regions of the world (Ramirez et al, 2013 and Sumathykutty, 1999) Some conducted to find potential pharmaceuticals or pesticides, and over 90% of the literature focuses on compounds that are cytotoxic, antifungal, antitumor, fragrant, or otherwise useful to humans (Dyer and Aparna Palmer, 2004; Lopez et al, 2002; Danelutte et al, 2003 ).
Piper aduncum widely spread in Borneo (Kalimantan), especially in Malinau District (Padmanaba and Sheil, 2014). P. aduncum, locally known Siri hutan, is a shrubs plant. It was potential as active against Aedes aegypti (Sudrajat, 2010); Marques, A.M. and Kaplan, M.A.C., 2015), fever medicine (Munawaroh et al, 2009), cytotoxic and antibacterial (Orjala, 1994). In Indonesia medicine traditional system, it was used as itching drug.The purpose of the present study was to investigate chemical compounds, toxicity and antioxidant activity from woods and bark ethanol extractsof Piper aduncum.
MATERIALS AND METHODS.
Equipment
The equipment used in this study is a set of distillation equipment, analytical
balance, blender, glass beaker, Erlenmeyer, measuring cups, funnels the stand, clamps, separating funnel, test tubes, pipette volume, pipette, micropipette 100-1000 ml size, flask, rod stirrer, light bulbs, hot plate. Concentrating the extract under reduced pressure using a rotary evaporator. determination of chemical constituents of extract using a Shimadzu GCMS QP2010 instrument models.
Material
The wood of Sirih hutan (Piper aduncum) was collected from Sempaja Samarinda city, East Kalimantan, Indonesia. The plant was identified by staff at Physiology Laboratory Mulwarman University, Samarinda. Ethanol was distilled before use in the maceration. Other materials used in this study is Whatman filtered and concentrated to under pressure using rotary evaporator. Total extract of wood bark was found 1 gram.
13 split. Flow control mode: Liner Velocity,
pressure: 80.2 kPa dengan total flow: 265.7 mL/min., Column Flow: 1.31 mL/min, Linear Velocity: 41.7 cm/sec, Purge Flow: 3.0 mL/min, Split Ratio: 200.0.
Antioxidant Assay
Determination of the antioxidant activity of Sirih hutan extracts was conducted using DPPH free radical scavenging. 1 mL of the extract (10; 25; 50; 75 and 100 ppm), then put into a test tube, respectively. Each tube was inserted 1 mL of DPPH 0.024 mg/mL, homogenized and left for 30 minutes in a dark place. All tubes were incubated in an incubator at 37 °C for 30 minutes before the absorbance was
measured using a UV-vis
spectrophotometer at wavelength maxima (λ) 515 nm. Vitamin C (0.01%; 0.02%; 0.03%; 0.04% and 0.05%) was used as a positive control. The antioxidant activity of the extract and vitamin C was determined by the formula:
%AA= Control Absorbance – Sample Absorbance x 100% Control Absorbance
RESULT AND DISCUSSION
The chromatograph of the Ethanolic extract of wood of Sirih hutan showed 11 compounds (see table 1). Five major
components namely Acetic acid (28.80%), followed by Nitrogen oxide (17.89%), 2-butanone (15.37%), monomethyl ester (CAS) Oxalic acid (10.68%), and 1, 2-Cyclopentanedione (8:38%). Then other minor components, namely, Ehanol (CAS) Ethyl alcohol (6.73%), 1-hydroxy-2-propanone (4:34%), 2,6-dimethoxy- Phenol (2:34%), propanoic acid, 2-oxo-, methyl ester (0.74% 0, and the last toloena (0.73%). Some minor components of the ethanolic extract have interesting biological activity based on of previous report. Compound 9 (1.2-Cyclopentanedione) is the precursor to form a 3-metyl-1,2-cycloptentanedione (3-MCP). 3-MCP may have a potential to minimize inflammation (Choi e al, 2007). Compound 10 (2,6-dimethoxy- Phenol) is widely used as a substrate (Wan et al, 2008), and it is a phenolic compound that is extensively used for the measurement of laccase activity, but is Often not exploited for its potential as an antioxidant compound (Adelakun et al, 2015). While the compound 11 (Dillapioda) known to have a very high gastroprotective activity (Marques, 2015). Dillapiolle combined with miristicin will show a synergistic effect on the antibacterial activity (Brazao et al, 2014)
Table 1. Chemical compounds of ethanolic exract by GC-MS analysis results
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4 1.417 15.37 methyl ethyl ketone 72 EtCOMe
5 1.521 28.80 Acetic acid 60 CH3COOH
6 1.757 4.34 1-hydroxy-2-propanone 74 CH3COCH2O
H
7 2.259 0.73 toloena 92 C6H5CH3
8 2.352 0.74 Propanoic acid, 2-oxo-,methyl ester
102 MeCOC(O)O
Me 9 3.750 8.38 1,2-Cyclopentanedione 98
10 10.705 2.34 2,6-dimethoxy- Phenol 154
11 14.900 4.00 Dillapiole 222
The antioxidant activity of ethanolic extract of Sirih hutan wood was measured by using DPPH radical scavenging method. DPPH radical will bond one atom of compound (in extracts) into a
1,1-diphenyl-2-picrylhydrazyl. This reaction occurs was characterized by a color change of purple to yellow. The following is the reaction mechanism DPPH radical scavenging by phenolic compound.
N N
+
O
OH H
NH N
+ O
O H
+ DPPH
-H-DPPH
NH N
+
O
O
15 The result of free radical DPPH
scavenging assay of ethanol extract of wood of sirih hutan with using vitamin C as a
positive control can be seen in table 2 below:
Table 2. The results of antioxidant activity IIC50 with DPPH radical scavenging method
No Sample IC50 (ppm)
1 Vitamin C 12.30
2 Ethanolic extract of Sirih hutan wood 259.74
IC50 value of the ethanolic extract of the wood of Sirih hutan was 259.74 ppm, it showed that the extract had a weak antioxidant properties (Jun et al, 2003). Weak antioxidant properties of the extract is supported by the results of the GC-MS analysis where only 10 and 11 compounds that have the potential as an antioxidant activity with a 6.34% total peak area.
CONCLUSSION
The content of the chemical compounds ethanolic extract of wood of
Sirih hutan based on GC-MS spectrum results indicate there are 11 compounds. Compound 11 (2,6-Dimethoxy-phenol) and 12 (Dillapiole) may be an antioxidant with a total peak area is 6.34%. So based on test results DPPH radical scavenging antioxidants are classified as weak.
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