PLANT (Scorodocarpus borneensis Becc. (OLACACEAE)
Wiwi Winarti1*), Rudy Kartika2, Partomuan Simanjuntak1,3
1Faculty of Pharmacy Pancasila University, Jakarta 12640, Indonesia
2F-MIPA Mulawarman University, Samarinda
3Puslit Biotechnology, LIPI Cibinong 16911
*) email : [email protected].
ABSTRACT
Bawang hutan plant (Scorodocarpus borneensis Becc) is one of Olacaceae family which contains polysulphide compound like as in Allium species plants. The aims of research is to know comparison activities (antioxidant, antibacteri, and toxicity) of ethyl acetate extract of bawang hutan plant.
Extraction is carried out by reflux and maceration with methanol and then portioned to ethyl acetate,n- BuOH and water. All extracts are tested to antioxidant (free radical scavenging method); antimicrobes (diffusion agar method) and toxicity (Brine Shrimp Lethality test). The result showed that ethyl acetate extract of stem bark (reflux and maceration) gave activities as anti oxidant are IC50 39,66 g/mL and 87,08 g/mL, respectively. While, ethyl acetate extracts of leaves (reflux and maceration) had IC50
182,90 ug/mL and 1145 ug/mL. The anti microbe showed ethylacetate of stem bark (reflux) inhibited growth of Staphylococcus aureus, and ethyl acetate extract of stem bark and leaves (all racemarion) inhibited growth of E. coli, and the stem bark of ethyl acetate (reflux) has the highest toxicity than others edtracts (LC50 40.53 ppm).
Key words: : bawang hutan; Scorodocarpus borneensis Becc.; Olacaceae; antioxidants; antimicrobial, toxicity (BSLT)
INTRODUCTION
Bawang hutan (Scorodocarpus borneensis Becc) is one of plant that can be used as drug. The study of literature, in native region of Sumatera and Kalimantan, these plants are widely used as a spice in cooking because the smell similar to garlic, so these plants contain compounds that are similar to polisulphide Allium species. Therefore, the plant is thought to have antioxidant activity and has a toxicity effect. Other parts of the plants that have been reported to be used as traditional medicine is the leaf that is as antidiarrheal, and contain flavonoids. While the results of the research before, it is known that ethanol extract of S. borneensis bark containing flavonoids and methylthiomethyl, methyl disulphide as having antimicrobial activity, and leaves of S. borneensis are suspected has the potential to produce compounds that have activities as an antioxidant, antimicrobial, and has the effect of toxicity. In this study conducted comparison of the activities of ethyl acetate extract (bark and leaves, extraction by reflux and maceration), includes testing of antioxidants with DPPH reagent, antimicrobial test with gelatin diffusion method against Gram-positive bacteria (Staphylococcus aureus) and Gram negative (Escherichia coli), and toxicity test using BSLT (Brine Shrimp Lethality Test). The cytotoxic properties shown in toxicity tests on larval shrimp Artemia salina Leach. with the observed death rate (mortality) of shrimp larvae.
METHODOLOGY Material
Materials research stem bark powder, dry simplicia (chopped) bark and leaves of bawang hutan (Scorodocarpus borneensis Becc.) collected from East Kalimantan
Comparison activities of………..
Chemicals and reagents
95% ethanol, methanol, ethyl acetate, n-butanol, n-hexane, DPPH (1,1-diphenyl-2-pikrilhidrazil), suspension of the test microbes (Staphylococcus aureus, Escherichia coli), Nutrient Agar (NA), to powder, peptone broth, salt water solution (NaCl), eggs Artemia salina Leach., dimethyl sulfoxide (DMSO) 1%, 1% cerium sulfate, silica gel 60, celite 545.
Method
Methanol extracts of stem bark and leaves are obtained by reflux and maceration. Simplicia dry stem bark and leaves 1 kg and 500 g respectively, refluxed at a temperature of 60o-70 ° C by using methanol solvent repeatedly (3 times), each for 3 hours and concentrated. The extracts partitioned to ethyl acetate, n-butanol, and
water. The next phase of ethyl acetate was collected and evaporated to obtain viscous extract of ethyl acetate, and then extracts ethyl acetate tested antioxidant activity with DPPH free radical method of arrest, antimicrobials by agar diffusion method, and toxicity tests with the method BSLT. Then the ethyl acetate extract had the highest activity class of chemical compounds identified by Farnsworth method.
RESULTS AND DISCUSSIONS Preparation of extracts
The weight of methanol extract of the bark is thick extract reflux is obtained 12.30 g (1.230%), and thick leaves extract 14.98 g (2.996%). The results of the manufacture of methanol extract of the bark maceration 5.60 g (1.867%), and the viscous extract 13.85 g (6.925%). Extraction results are presented in Table 1.
Table 1. The results of extraction of the bark and leaves of S.borneensis with the methanol Extraction process Methanol extract Weight, g (%) color
stem bark 12.30 (1.23) black brownish
leaves Reflux
14.98 (2.99) green blackish
stem bark 5.60 (1.867) black brownish
leaves Maceration
13.85 (6.925) green blackish
Partitioning results show that ethyl acetate extract of the stem bark from S. borneensis maceration and reflux are 3.05 g (0.305%) and 1.47 g (0.490%). While the ethyl acetate extract of leaves of 8.75 g (1.750%) and 7.93 g (3.965%). Extraction results are presented in Table 2
Table 2. The results of ethyl acetate extract partition stem bark and leaves of S.borneensis Extractionprocess Ethyl acetate extract Weight, g (%) color
Stem bark 3.05 (0.305) black
leaves Reflux
8.75 (1.750) green blackish
Stem bark 1.47 (0.490) black brownish
leaves Maceration
7.93 (3.965) green blackish
Ethyl Acetate Extract Activity Test Antioxidant test
Testing the antioxidant activity of ethyl acetate extract (stem bark and leaves) by reflux and maceration, with concentrations (100, 50, 25 ug / mL) showed that a sample has antioxidant activity when giving IC50 values < 200 ug / mL. The results of the fourth test extracts and positive control of vitamin are presented in Table 3.
Wiwi Winarti
Table 3. The test results free radical of the ethyl acetate extract & vitamin C
Samples Concentration (µg/mL) Inhibition (%) IC50 (µg/mL) Information
100 81.66
50 68.88
The Ethyl acetate extract of the stem
bark (Reflux) 25 32.79
39.66 most Active
100 56.26
50 32.03
The ethyl acetate extract the stem
bark (Maceration) 25 17.01
87.08 Active
100 26.60
50 12.12
The ethyl acetate extract of leaves
(Reflux ) 25 5.8
182.90 Active
100 4.94
50 3.19
The ethyl acetate extract of leaves
(Maceration) 25 1.64
1145.00 inactive
15 8.83
12 66.49
Vitamin C (positive control)
9 43.29
9.87 -
The results showed that the ethylacetate extract of stem bark (reflux or maceration) and ethyl acetate extracts of leaves (reflux), has a reduction potential of free radicals with IC 50 < 200 ug / mL.
There were significant differences between the the percent inhibition of ethyl acetate extract reflux and maceration, the percent inhibition of the ethyl acetate extract reflux was much larger compared with maceration.
Antimicrobial Test
Table 4. The test results of antimicrobial activities ethyl acetate extracts & chloramphenicol against the bacterium Escherichia coli
DIG (mm)
Microbe Test Samples Concentrati
on (µg/mL) I II III
1500 - - -
1000 - - -
Escherichia coli The ethyl acetate extract of the bark (Reflux)
500 - - -
1500 9 9 9,5
1000 - - -
Escherichia coli The ethyl acetate extract the bark (Maceration)
500 - - -
1500 - - -
1000 - - -
Escherichia coli The ethyl acetate extract of leaves (Reflux)
500 - - -
1500 7,5 7,8 8
1000 - - -
Escherichia coli The ethyl acetate extract of leaves (Maceration)
500 - - -
1500 23 23 23,3
1000 20 20 20,5
Escherichia coli Cloramphenicol (positive control)
500 18 18,6 19
Note DIG : Diameter of Inhibition Growth (-) : No inhibion
Comparison activities of………..
The test results of antimicrobial activity of ethyl acetate extract of the bacterium Escherichia coli are presented in Table 4. These results indicate that only the ethyl acetate extract of the stem bark and leaves of S.borneensis by maceration that have inhibition a concentration of 1500 ug / mL, while the stem bark and leaves extracts by reflux has no inhibition.
The results of antimicrobial activity of ethyl acetate extract against Staphylococcus aureus bacteria are presented in Table 5. The results showed that the ethyl acetate extract of the bark from S.borneensis (reflux) that have inhibition against of bacteria Staphylococcus aureus at concentrations 1500 ug / mL, whereas thee ethyl acetate extract of the leaves (reflux), stem bark and leaves by maceration has no inhibition against S. aureus bacteria.
Table 5. The results of antimicrobial activities ethyl acetate extracts & chloramphenicol against Staphylococcus aureus
IGD (mm) Microbe Test Samples Concentration (µg/mL)
I II III
1500 8 8,5 8,5
1000 - - -
Staphylococcus aureus
The ethyl acetate extract of the bark
(Reflux) 500 - - -
1500 - - -
1000 - - -
Staphylococcus aureus
The ethyl acetate extract the bark
(Maceration) 500 - - -
1500 - - -
1000 - - -
Staphylococcus aureus
The ethyl acetate extract of leaves
(Reflux) 500 - - -
1500 - - -
1000 - - -
Staphylococcus aureus
The ethyl acetate extract of leaves
(Maceration) 500 - - -
1500 27 27.3 27.8
1000 25 25 25.3
Staphylococcus aureus
chloramphenicol
(positive control) 500 22 22.6 23
Toxicity test (BSLT)
Table 6. LC50 value of the results BSLT ethyl acetate extract of the bark and leaves from S. borneensis extraction by maceration and reflux.
Ethyl acetate extract Concentration
(µg/mL) mortality (%) LC50
(µg/mL) information
1000 100.00
100 93.94
Stem bark (reflux)
10 9.38
40.53 most active
1000 100.00
100 69.70
Stem bark (Maceration)
10 7.50
63.67 active
1000 100.00
100 18.75
Leaf (reflux)
10 3.57
15.37 active
1000 97.78
100 40.54
Leaf (Maceration)
10 11.54
100.25 active
Wiwi Winarti
The results of the ethyl acetate extracts showed that the ethyl acetate extract of the stem bark from S. borneensis by (reflux) has the highest activity with LC50 value 40.53 ug / mL, followed by ethyl acetate extract of the stem bark in maceration with LC50 63.67 ug / mL, ethyl acetate extract leaves in the maceration with LC50 100.25 ug / mL and ethyl acetate extracts of S. borneensis leaves by refluxed with LC50 155.37 ug / mL (Table 6).
The observations of ethyl acetate extract has the effect due to its toxicity LC 50 < 1000 ug / mL.
From the above data, all ethyl acetate extract are toxic, and the highest activity are the ethyl acetate extract of the bark from S. borneensis extracted by reflux (LC50 40.53 ppm). The identification of chemical compound group showed that in the powder and ethyl acetate extracts contain flavonoid, steroid, triterpenoid coumarin and sulphide compound.
Table 7. The identification of chemical compound group for stem bark of S. borneensis
No. Chemical compounds Powder symplisia Extract of ethyl acetate (reflux)
1 Alkaloid - -
2 Flavonoid + +
3 Saponin + -
4 Tanin - -
5 Steroid/triterpenoid + +
6 Kuinon - -
7 Minyak atsiri - -
8 Coumarin + +
9 Sulphide + +
Information :
+: Give positive results +: Give negative results CONCLUSION
The result of the research comparison activities of leaves and bark of ethylacetat extract from bawang hutan plant (Scorodocarpus borneensis Becc) are:
1. The stem bark’s extract (reflux and maceration) has the highest antioxidant activity.
2. The stem bark and leaf’s extract (maceration) has inhibition against E. coli.
3. The stem bark’s extract (reflux) has toxicity against Artemisia larvae.
ACKNOWLEDGEMENT
The Authors thanks to Ms. Rinawati for assistance in this research.
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