The Effectiveness of Kirinyuh Leaves (Eupatorium odoratum L.) and
Allium chinense Extract against Staphylococcus aureus and
Escherichia coli
Saipul Sihotang
1
, Muhammad Usman
2
and Sartini
2
1
Faculty of Medicine, Universitas Prima Indonesia, Medan, Indonesia
2
Faculty of Science and Technology, Universitas Medan Area, Medan, Indonesia
Keywords: Ki rinyuh Leaves, Phytochemicals, Secondary Metabolites.
Abstract: This study aims to determine secondary metabolite compounds of kirinyuh leaf extract (Euphatorium
odoratum L) and hobo (Allium chinense and their antibacterial properties against Staphylococcus aureus and
Escherichia coli. The research consisted of three steps, the tirst was plant extraction and maceration using
methanol as a solvent. The second, phytochemical screening to confirm secondary metabolites (alkaloids,
flavonoids, terpenoids and steroids) The third, antibacterial test against Staphylococcus aureus and
Echerichia coli at concentrations 0, 1, 5, 10, 15, 20 and 25 %, clyndamicin used as positive control. Data
were analyzed using SPSS Version 23 software and continued statistically by Duncan Multiple Range Test.
Results showed that secondary metabolites such as alkaloids, flavonoids, steroids and saponins were
detected in kirinyuh. Whereas, secondary metabolites of hobo contain flavonoids, steroids and saponins.
Extract concentration 25% was the most effective for kirinyuh in inhibiting Staphylococcus aureus and
Escheriachia coli with the highest inhibition 15.00 to 19.03 mm, whereas, the inhibition of hobo extract
effective in inhibiting S. aureus 12.40 mm to 15.20 mm.
1 INTRODUCTION
The use of plants as traditional medicines become
increasingly widespread. One of the benefits of
medicinal plants as an antimicrobe is no side effect
and potential to reduce infection caused by bacteria,
fungi, and parasites. The used of antibiotic to reduce
diseases caused by pathogenic bacteria can cause
bacterial resistance (Awoyinka et al. 2007).
Therefore, the used medicinal plants as an
alternative to reduce bacterial infection. Medicinal
plants that can be used to treat infectious diseases
are kirinyuh (Euphatorium odoratum L.) and hobo
(Allium chinense).
In Thailand kirinyuh was used as a wound
medicine, coagulant, and as an antiseptic (Irobi,
1997), in Nigeria used as a therapy for malaria
(Rungnapa, 2003), while in Indonesia the plant used
as a medicine for new wounds, fever, cough, and to
stop the disease. bleeding (Purwati, 2003). Even so,
this plant is still very rarely used by the people of
Indonesia because it is considered a nuisance plant
that is difficult to eradicate.
Inya-agha et al. (1987) reported that E. odoratum
leaf extract contains tannins, phenols, saponins, and
its essential oil contains α-pinene, cadinene,
camphore, limonene, β-caryophyllene, cadinol
isomers. Thakong (1999) reported that chloroform
extract from E. odoratum leaves showed high
activity against chloroquin-resistant Plasmodium
falciparum (K1). The compound isolated from the
chloroform extract fraction of E. odoratum leaves
was isosacuranetin, which was inactive against P.
falciparum at a maximum concentration of 5 μg / ml.
The ethanol extract from the leaves of E. odoratum
showed antibacterial activity against Pseudomonas
sp., Escherichia coli, B. thuringensis, Klebsiella sp.
and Streptoccocus faecalis (Irobi, 1997).
Allium chinense is the onion class commonly
used for seasoning and flavoring dishes. Batak
chives or onions contain nutrients that can prevent
cancer and hypertension. This plant contain
compounds as antioxidants, antibiotics, anti-cancer,
and antibacterial agents (Rudi, 2012). The aims of
the present study was to determine composition
secondary metabolites crude extract of kirinyuh
566
Sihotang, S., Usman, M. and Sartini, .
The Effectiveness of Kirinyuh Leaves (Eupatorium odoratum L.) and Allium chinense Extract against Staphylococcus aureus and Escherichia coli.
DOI: 10.5220/0010613500002775
In Proceedings of the 1st International MIPAnet Conference on Science and Mathematics (IMC-SciMath 2019), pages 566-570
ISBN: 978-989-758-556-2
Copyright
c
 2022 by SCITEPRESS – Science and Technology Publications, Lda. All rights reserved

leaves and Allium chinense as anti-microbial against
Staphylococus aureus and Escherichia coli.
2 MATERIALS AND METHOD
2.1 Sample Collection
The materials used in this study were E. odoratum
and Allium chinense which were obtained from
Tanah Karo area (Brastagi), North Sumatera.
Staphylococcus aureus, and Escherichia coli were
obtained from Biology Laboratory, Department of
Biology, Medan Area University.
2.2 Extraction
The procedure used in this study consisted of three
steps, the first was the extraction of kirinyuh
(Euphatorium odoratum L) and hobo (Allium
chinense) by maceration using methanol as solvent.
The second was phytochemical screening by
identifying secondary metabolites (alkaloids,
flavonoids, terpenoids and steroids). The third,
bioactivity test against Staphylococcus aureus and
Eschericia coli using extract concentrations 0, 1, 5,
10, 15 , 20 and 25%. Extraction was started by
maceration using n-hexane and methanol as solvents
for 3 × 24 hours each with a solvent every 24 hours.
The extract was then concentrated using rotavapor.
2.3 Phytochemical Test
Phytochemical screening for alkaloids, flavonoids,
saponins, phenolics, triterpenoids and steroids was
detected using reagents that specific for each
compound. Screening for alkaloids using reagents
such as Meyer, Bouchardat, Wagner and
Drangendorff. Screening for flavonoid was
conducted according to Harbone (1996). The
reagents used consisted of NaOH solution, sulfuric
acid (H
2
SO
4
), aqueous Mg-HCl solution, the
reaction will form a blue violet color, with
concentrated (H
2
SO
4
) will form a yellowish orange
color, and with a dilute Mg-HCl solution with pink
color. The presence of color by the addition of the
reagents indicate the presence of flavonoid.
Screening of triterpenoids and steroids was carried
out by the Lieberman-Burchard adhesion (Harbone,
1996). The presence of triterpenoids was indicated
by the presence of a red color change, while blue or
purple indicates the presence of steroids. Phenolic
compounds were screened using 1% FeCl
3
reagent
(Harbone, 1996). The appearance of blue or purple
blue indicates positive for phenolic. Saponin
compound screening was carried out by using boiled
water in a test tube and then shaking it vigorously
for a while. If a permanent foam is formed for about
15 minutes with the addition of one or two drops of
2 N hydrochloric acid (HCl), it shows a positive test
for saponins (Harborne, 1996).
2.4 Antimicrobial Test
To examine antimicrobial activity, extract of
Euphatorium odoratum leaves and hobo (Allium
cinense) were diluted in sterile distilled water with
concentrations of 1, 5, 10, 15 and 20%. The bacterial
culture suspension was taken from existing cultures
in the laboratory. The bacterial suspension then were
inoculated by spread method in petri dishes (9 cm in
diameter). Five paper discs for each petri dish. Each
of the disc then was dripped for each extract
concentration. Petri dishes were divided 4 parts
based on the concentration. All cultures were
incubated at 37
o
C for 24 hours. The presence of
clear zone around the plant extract were observed.
The diameter of the inhibition zone was determined
using calipers according to the Kirby-bauer of
Susceptibility Testing method in mm (Cappucino &
Sherman, 1999).
2.5 Data Analysis
This study used a completely randomized factorial
design with experimental methods. Data collected
were analyzed using SPSS 23, followed by Duncan
Multiple Range Test (DNMRT) for significant
differences among treatments given.
3 RESULTS AND DISCUSSION
3.1 Phytochemical Screening
Secondary metabolites of kirinyuh leaf can be seen
in Table 1. The metabolites found were flavonoids,
alkaloids, and saponins. Whereas, secondary
metabolites of Allium chinense was shown in Table
2.
The Effectiveness of Kirinyuh Leaves (Eupatorium odoratum L.) and Allium chinense Extract against Staphylococcus aureus and
Escherichia coli
567

Table 1: Phytochemical test of methanol extract of
kirinyuh leaves.
No
Compound
identification
Reactor Result
1 Flavonoid
FeCl
3
1%
+
Mg-HCl
-
NaOH 10%
-
H
2
SO
4
-
2 Alkaloid
Dragendorf
+
Bouchardat
+
Meyer
+
Wagner
+
3
Steroid/
Terpenoid
Salkowsky
+
CeSO
4
1% +
H
2
SO
4
10%
+
4 Saponin
Sample + H
2
O +
HCl 2N
+
Table 2: Phytochemical test of methanol extract of Allium
chinense
No
Compound
identification
Reactor Result
1 Flavonoid
FeCl
3
1%
+
Mg-HCl
+
NaOH 10%
-
H
2
SO
4
-
2 Alkaloid
Dragendorf
-
Bouchardat
-
Meyer
-
Wagner
-
3
Steroid/
Terpenoid
Salkowsky
+
CeSO
4
1% +
H
2
SO
4
10%
+
4 Saponin
Sample + H
2
O +
HCl 2N
+
3.2 Antibacterial Test
Anti-bacterial tests were carried out to determine the
ability of methanolic extract of kirinyuh and hobo in
inhibiting growth of Staphylococcus aureus and
Escherichia coli. The effectivity both of the extracts
was indicated by the presence of a clear zone around
the discs that contain plant extract (Table 3).
Table 3: Diameter of inhibition zone (mm) of methanol
extract of kirinyuh leaves with various concentrations (%).
Treatments Staphylococcus aureus Escherichia coli
(+) Control
16.97
a
19.13
a
(-) Control
0.00
f
0.00
d
1%
2.67
e
6.67
c
5%
5.00
de
6.00
c
10%
6.33
de
11.33
bc
15%
8.33
d
15.33
b
20%
13.00
c
16.67
ab
25%
15.00
ab
19.03
a
Numbers followed by the same letters not significantly different
(0.5) according to Duncan New Multiple Range Test
Table 4: Diameter of inhibition zone (mm) methanol
extract of A. chinense with various concentrations (%).
Treatment Staphylococcus aureus Escherichia coli
(+) Control
16.50
a
17.50
a
(-) Control
0.00
f
0.00
g
1%
5.00
d
9.00
f
5%
7.00
c
11.00
e
10%
8.00
bc
13.00
d
15%
10.00
b
13.40
cd
20%
12.00
b
14.00
bc
25%
12.40
b
14.50
b
Numbers followed by the same letters not significantly different
(0.5) according to Duncan New Multiple Range Test
Table 3 shows that the concentration of 25% is
the best treatment to S. aureus and hobo extracts to
S. aureus and E. coli with inhibition f 15.00 mm and
19.03 mm. Table 4 shows that the concentration of
25% is the best treatment of giving S. aureus and
hobo extracts to Staphylococcus aureus and
Escherichia coli with an average inhibition 12.40
mm to 14.50 mm.
Kirinyuh leaf extract (Figure 1) and hobo (Figure
2) showed positive effect in inhibiting S. aureus and
E. coli. It can be seen that 25% treatment is the best
treatment of kirinyuh leaf extract and hobo against S.
aureus and E. coli with the average inhibition power
data of 15.00 mm and 19.03 mm and 12.40 mm and
15.20 mm. Chemical compounds in kirinyuh leaf
extract and raw hobo such as flavonoids, alkaloids,
steroids and saponins have antibacterial activity.
According to Qin and Sihotang (2020) and
Simanullang et al. (2021) that secondary metabolite
compounds such as flavonoids, alkaloids and
saponins have potential as antimicrobial agents.
IMC-SciMath 2019 - The International MIPAnet Conference on Science and Mathematics (IMC-SciMath)
568

Figure 1: The percentage inhibition of bacterial growth in
kirinyuh leaf extract.
Figure 2: The inhibition zone bacterial growth on hobo
extract.
According to Andriani et al., (2016) Secondary
metabolite compounds such as flavonoids, alkaloids
and saponins have potential as antimicrobial agents.
This was confirmed by Heinrich (2009) who stated
that flavonoid compounds.
Kurnonealoin compounds can cause bacterial
proteins to become inactive and lose their function,
while saponins can dissolve lipids in bacterial cell
membranes, as a result it can reduce lipid tension,
change cell permeability, abnormal cell function and
eventually lysis and cause death (Ismiyati, 2014). S.
aueus and E. coli are pathogenic bacterial isolates
with thick cell walls because they contain a lot of
peptidoglycan and are quite thick (20 -80 nm) and
also contain teichoic acid and lipoteichoic acid
(Heinrich et al. 2009).
This bacterial cell wall arrangement contains
only one layer of plasma membrane, this is what
causes its osmotic pressure to drop dramatically
when given the anthracurnonealoin complex
contained in aloe vera extract. So that the bacterial
cell will have difficulty controlling the respiration
process and ion transport from outside the cell.
Based on the results of data analysis from the One-
Way Anova test, the significance results were
smaller than 0.05, which means that there were
significant differences in the inhibitory power of
various concentrations of kirinyuh leaf extract and
hobo raw on the growth of S. aureus and E. coli
isolates in compared to control (-) and ( +), meaning
that the three concentrations of kirinyuh leaf extract
and hobo have antibacterial effects against S. aureus
and E. coli but not as strong as the control (+).
4 CONCLUSION
Secondary metabolites of kirinyuh and hobo is a
group of flavonoids, alkaloides, steroids and
saponins that potential as antibacterial to inhibit S.
aureus and E. coli.
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