PHYTOCEMICAL ANALYSIS AND ANTI OKSIDANT ACTIVITY OF THE WOOD ETHANOLIC EXTRACTS OF SIRIH HUTAN (Pipe aduncum)

  

PHYTOCEMICAL ANALYSIS AND ANTI OKSIDANT ACTIVITY OF THE

WOOD ETHANOLIC EXTRACTS OF ₁ SIRIH HUTAN (Pipe aduncum) Erwin

  

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 IC = 259,74 ppm. ₅₀ 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

  IC ^{50 259.74 ppm.} Keyword: Sirih hutan, antioxidant activity, and GC-MS

  1

  INTRODUCTION

  Extraction

  C, Injection Mode:

  o

  C , injection temp. 280.00

  o

  Ethanolic extract wood of Sirih hutan analyzed by GC-MS using a Shimadzu GC-MS model QP2010 instrument in a system operated dengan electron impact yang digunakan dalam 70 kV dan column Oven temp. 60.0

  GC-MS Analysis

  were macerated with ethanol at room temperature and repeated 4 times, filtered and concentrated to under pressure using rotary evaporator. Total extract of wood bark was found 1 gram.

  Sirih hutan

  149 gram dried powder of wood of

  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 filter paper no. 1, aluminum foil, DPPH (2,2-diphenyl-1-picrylhidrazyl) and Vitamin C.

  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

  Material The wood of Sirih hutan (Piper aduncum ) was collected from Sempaja

  Concentrating the extract under reduced pressure using a rotary evaporator. determination of chemical constituents of extract using a Shimadzu GCMS QP2010 instrument models.

  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.

  Equipment

  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 extracts of Piper aduncum.

  aduncum, locally known Siri hutan, is a

  Borneo (Kalimantan), especially in Malinau District (Padmanaba and Sheil, 2014). P.

  Piper aduncum widely spread in

  300 species of piper are endemic to Southeast Asia, including the East Indian islands and northern Australia. Only two species are native to Africa. Most Piper species grow in wet, warm, low land rain forests. Many species of Piper have been 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 ).

  al , 2013 and Sumathykutty, 1999) Some

MATERIALS AND METHODS.

  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)

  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

  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.

• – Sample Absorbance x 100% Control Absorbance

RESULT AND DISCUSSION

  Table 1. Chemical compounds of ethanolic exract by GC-MS analysis results peak Retention time

  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%).

  Compound Molecular Weight

  Molecular Formula 1 1.314

  17.89 Nitrogen oxide

  44 N2O 2 1.340 10.68 monomethyl ester (CAS)

  Oxalic acid 104 MeOC(O)COO H 3 1.380

  6.73 Ethyl alcohol

  46 EtOH

  Peak area (%)

  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}

  Figura 1. reaction mechanism DPPH radical scavenging by the phenolic compound

• – 526

  Sirih hutan based on GC-MS spectrum results indicate there are 11 compounds.

  64: 555-559 5. Dyer, L.A. and Aparna Palmer, D. N. 2004, Piper : A Model Genus for Studies of Phytochemistry, Ecology, and Evolution, Kluwer Academic/Plenum Publishers, New York Boston, Dordrecht, London, Moscow.

  crassinervium Kunth. Phytochemistry,

  4. Danelutte A, Lago J, Young M, Kato M 2003. Antifungal flavanones and prenylated hydroquinones from Piper

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  2. Brazao, M. A.B., Brazao, F.V., Maia, J.G.S., and Monteiro, M.C., 2014, Antibacterial activity of the Piper aduncum oil and dillapiole, its main constituent, against multidrug-resistant strains, Boletín Latinoamericano y del

   uly 2015

  University of Pretoria, Pretoria 0002, SouthAfrica

  Enzymatic modification of 2,6- dimethoxyphenol for the synthesis of dimers with high antioxidant capacity, Research and Postgraduate Studies ,

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  REFERENCE

  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.

  The content of the chemical compounds ethanolic extract of wood of

  CONCLUSSION

  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.

  50 value of the ethanolic extract of

  IC

  2 Ethanolic extract of Sirih hutan wood 259.74

  12.30

  1 Vitamin C

  50 (ppm)

  IC

  with DPPH radical scavenging method No Sample

  

50

  Table 2. The results of antioxidant activity IIC

  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:

• –1619

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• –2122.

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