Nur
Patria Tjahjani1*, Candra Aini Roro Rias2
1,2Akademi Kesehatan 17
Agustus 1945, Semarang, Indonesia
Email: nurpatriacahyani@gmail.com
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KEYWORDS |
ABSTRACT |
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Escherichia
coli, Bay Leaf Extract, Secang Wood Extract |
Bay leaves (Syzygium polyanthum) and a second wood
(Caesalpinia sappan L) are widely known to the public as natural antiseptics
for prevention and treatment, including treating digesting diseases, one of
which is diarrhea caused by Escherichia coli. Bay leaves and Secang wood can
be used as antiseptics because they contain the active substance tannin. This
research aims to determine the difference in the antibacterial effectiveness
of 96% ethanol extract of bay leaves (Syzygium polyanthum) and 96% ethanol
extract of second wood (Caesalpinia sappan L) on growth Escherichia coli
bacteria. The type of research is experimental. The method used in this
research is the Kirby-Bauer diffusion method. This method observes the clear
zone produced from a blank disk that has been treated with bay leaf extract
and second wood extract at a concentration of 30%, 60%, and 90% for
incubation at 37⁰C for 24 hours. From the research results, it can be
concluded that the ethanol extract of bay leaves with a concentration of 30%,
60%, and 90% with an inhibition zone diameter of 0 mm, 7,6 mm, and 9,6 mm.
The second wood at a concentration of 30%, 60%, and 90% with an inhibition
zone diameter of 10 mm, 11,3 mm, and 14 mm. The statistical test shows that
the data is normally distributed with a parametric test ANOVA, p =0,00, which
means that there is no significant difference in the diameter of the
inhibition zone between the ethanol extract of bay leaves and the ethanol
extract of second wood. |
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DOI: 10.58860/ijsh.v3i10.257 |
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Corresponding Author: Nur Patria Tjahjani*
Email: nurpatriacahyani@gmail.com
INTRODUCTION
Diarrhea is a
clinical condition characterized by increased frequency and decreased stool
consistency when compared to normal individuals. In patients with diarrhea, the
frequency of defecation occurs more than three times within 24 hours with a
mushy or liquid stool consistency
Along with
the development of science and technology, people now prefer to utilize natural
remedies rather than chemical-based remedies. The utilization of plants as
medicinal plants is chosen because these materials have relatively low side
effects. In addition, medicinal plants are much more affordable than chemical
treatment.
Indonesia is
rich in traditional medicinal plants. Traditional medicinal plants are known or
believed to have properties as raw materials for traditional medicine. One of
the traditional medicinal plants that is often used by the community is bay
leaf. This bay leaf can also be used as an antibacterial for diarrhea,
hypertension, diabetes, and gout. Bay leaves contain substances that are
efficacious as antimicrobials, namely tannins, essential oils, flavonoids,
alkaloids, and saponins
Sappan wood
is one of the plants that is currently not fully utilized by the community.
Sappan wood has properties in treating several diseases. One of them is
antidiarrheal
Escherichia
coli is a rod-shaped, gram-negative bacterium that is a normal inhabitant in
the digestive tract of humans and animals. However, Escherichia coli is also
one of the bacteria that cause diarrhea
On sappan
wood extract granules (Caesalpinia sappan L.) and temu mango rhizomes (Curcuma
mango Val & Zijp.) as antibacterial shows sappan wood extract granules with
concentrations of 12.5%, 25%, 50% and 100% can inhibit Escherichia coli,
Salmonella sp and Shigella sp bacteria. Research by Andri Wahyu Saputri, et al
Based on the
description above, the researcher is interested in conducting research on
differences in the inhibitory concentration of ethanol extracts of bay leaves
and second wood against the growth of Escherichia coli bacteria at
concentrations of 30%, 60%, and 90%.
This study
aims to address the gap in research by comparing the inhibitory concentrations
of ethanol extracts of bay leaves and sappan wood on Escherichia coli at
specific concentrations of 30%, 60%, and 90%. This comparative analysis
provides insight into the efficacy of these traditional plants at different
extract concentrations, contributing novel findings on optimal concentrations
for antibacterial activity. This research could expand options for affordable,
plant-based diarrhea treatments, enhancing the scientific basis for traditional
Indonesian medicine.
METHOD
This study is an experimental study to see the
difference in each concentration of ethanol extract of bay leaves (Syzygium
polyanthum) and ethanol extract of second wood (Caesalpinia sappan L.) against
the growth of Escherichia coli bacteria. This research was conducted at the
Chemistry Laboratory and Microbiology Laboratory of the Health Academy on 17
August 1945 Semarang. Bay leaves, and sappan wood are sold at Dami Sariwana
Herbal and Simplisia Store. Bay leaves, and secang wood that have become 600
grams of simplisia are sold at Toko Jamu and Simplisia Dami Sariwana.
The data collected are primary data obtained from
the measurement of the inhibition zone and each concentration of bay leaf
extract and secang wood with the Kirby-Bauer disc method against Escherichia
coli bacteria in the laboratory.
Data Analysis
Data from the
study of the effect of bay leaf extract and secang wood on Escherichia coli
were analyzed using the SPSS program to see if there were significant
differences in the effectiveness of each test disc containing negative control,
various concentrations of bay leaf extract, secang wood and positive control in
inhibiting the growth of Escherichia coli.
From the test
results, data is obtained, which is made with a table diagram, and if the data
obtained is normally distributed, then a statistical test is carried out with
the ANOVA test and continued with the T-test. If the data is not normally
distributed, then use a non-parametric test, namely the Mann-Whitney test.
RESULT AND DISCUSSION
The results of
maceration of bay leaves (Syzygium polyanthum) and secang wood (Caesalpinia
sappan L.) with 96% ethanol solution obtained yields the results can be seen in
the table below:
Table 1.
Extract Weighing Results
and Yield Salam Leaf and Secang Wood Using 96% Ethanol Solvent
|
Simplisia
Name |
Simplisia
Weight (gram) |
Extract
Weight (gram) |
Yield
Value (%) |
|
Bay Leaves |
300,0 |
27,09 |
9,03 |
|
Secang
Wood |
300,0 |
25,33 |
8,44 |
The test
results on the activity test of 96% ethanol extract of bay leaves (Syzygium
polyanthum) and 96% ethanol extract of secang wood (Caesalpinia sappan L.)
against the growth of Escherichia coli bacteria that have been grown in Petri
dishes using Mueller Hinton Agar media apparently show different abilities in
each concentration given can be seen in the table below:
Table 2.
Results of Measurement of
Inhibition Zone Diameter of 96% Ethanol Extract of Salam Leaf Against
Escherichia coli Bacteria
|
Extract
Type |
Concentration
(%) |
Inhibition
Zone Diameter (mm) |
|
Bay
Leaves |
30 |
0,00 |
|
|
60 |
7,60 |
|
|
90 |
9,60 |
|
|
Control
+ |
25,00 |
|
|
Control
- |
0 |
Table 3.
Measurement Result of
Inhibition Zone Diameter of 96% Ethanol Extract of Secang Wood Against
Escherichia coli Bacteria
|
Extract
Type |
Concentration
(%) |
Inhibition
Zone Diameter (mm) |
|
Secang
Wood |
30 |
10,00 |
|
|
60 |
11,30 |
|
|
90 |
14,00 |
|
|
Control
+ |
25,00 |
|
|
Control
- |
0 |
Diagram of Measurement Results
of Inhibition Zone Diameter of Salam Leaf and Secang Wood
Data Analysis
Research data
in the form of concentrations and differences in bay leaves and secang wood
against Escherichia coli growth were analyzed descriptively using average
values, then presented in tables. Furthermore, it was analyzed statistically,
and it was shown that the data was normally distributed using the Saphiro-Wilk
test, which was continued with the ANOVA test. The significance value in this
study uses the variables analyzed at a value of p> 0.05.
The results of
the ANOVA test on the three concentration groups of bay leaf ethanol extract
and second wood ethanol extract against Escherichia coli bacterial growth
showed no difference in the inhibition zone between bay leaf ethanol extract
and second wood ethanol extract against Escherichia coli bacterial growth with
the analyzed significance value of p = 0.00 <0.05.
Hypothesis
Hₒ: There is no
difference in the zone of inhibition between the ethanol extract of bay leaves
and ethanol extract of secang wood against the growth of Escherichia coli
bacteria.
H1: There is a difference in
the inhibition zone between the ethanol extract of bay leaves and ethanol
extract of secang wood against the growth of Escherichia coli bacteria.
The results of
the study using the Kirby-Bauer Diffusion method showed that 96% ethanol
extract of secang wood proved to be stronger in inhibiting the growth of
Escherichia coli bacteria than 96% ethanol extract of bay leaves in various
concentrations, which can be seen in Figure 3 Diagram of the Inhibition Zone
Diameter Measurement Results of Bay Leaves and Secang Wood.
The greater
the concentration, the greater the inhibition against the growth of Escherichia
coli bacteria. Bay leaf extracts with concentrations of 30%, 60%, and 90% with
inhibition zone diameters of 0 mm, 7.6 mm, and 9.6 mm, while sappan wood at
concentrations of 30%, 60% and 90% with inhibition zone diameters of 10 mm;
11.3 mm and 14 mm indicate that both bay leaf extracts and sappan wood have
antibacterial abilities but there are differences in the inhibition zones
produced in bay leaf extracts with a concentration of 30% cannot inhibit
Escherichia coli bacteria because there is no clear zone around the paper disk.
There is a
difference in the ability to inhibit the growth of Escherichia coli bacteria
between young bay leaves and old bay leaves. The results of research by Mufti
Kharismawati, et al.
Research
conducted by Andri Wahyu Saputri et al.
Stated that
bay leaves contain tannin, alkaloid, flavonoid, and saponin compounds that can
inhibit Escherichia coli bacteria. Research conducted by Indrias Cahyayuliani,
et al.
The
phytochemical screening of bay leaves conducted by Arum Samudra
Tannins are
active secondary metabolite compounds that are known to have several properties,
including antidiarrheal, astringent, antibacterial, and antioxidant
This research
is in line with research conducted, which proves that secang wood extract
(Caesalpinia sappan L.) is able to inhibit the growth of Escherichia coli
bacteria at a concentration of 12.5%; 25%; 50% and 100% with an inhibition zone
diameter of 8 mm; 8 mm; 13.3 mm and 16.2 mm. The results of Susanto's research,
stated that the content of tannin phytochemical compounds in ethanol extract of
secang wood was 10.42%, and in water extract of secang wood was 5.95%.
Sappan wood
plants have the chemical content of tannins, brazilin, alkaloids, saponins,
flavonoids, phenyl propane, and terpenoids. Sappan wood contains phenolic
compounds such as flavonoids, which have free radical-catching antioxidant
activity. Anthocyanins are one of the flavonoid groups in secang wood.
Anthocyanin compounds are compounds that are good for health because they have
antioxidant activity
Escherichia
coli is an opportunistic germ that is widely found in the human colon, and as a
normal colon flora, the nature of Escherichia coli can cause primary infection
in the colon so that it can cause diarrheal disease
The active
compounds in bay leaves, besides tannins, also have flavonoids as antibacterial
active compounds with the mechanism of action of forming complex compounds with
extracellular and soluble proteins so as to damage the bacterial cell membrane,
followed by the release of intracellular compounds. The structure of the cell
wall and bacterial cytoplasmic membrane becomes unstable because the structure
of the bacterial cell protein is damaged. As a result, the cell permeability
function is disrupted, and the bacterial cell will experience lysis, which
results in the death of the bacterial cell.
Sappan wood
contains saponins that increase membrane permeability, which causes cell
hemolysis; if saponins interact with bacterial cells, the bacteria will be
lysed
This study
uses a simple extraction method, namely maceration using simplisia made by
soaking because this process does not use the heating stage, which causes the
active substance content to be damaged, simple equipment, costs are not too
expensive and easy to work on. In principle, extraction is to dissolve and
withdraw compounds using the right solvent
Maceration of
bay leaves and secang wood using 96% ethanol. Ethanol 96% has a boiling point
of 78.4⁰C; ethanol has the ability to distill with a wide polarity
ranging from non-polar to polar compounds. The selection of 96% ethanol solvent
is based on the compounds to be taken.
Ethanol is also non-toxic and harmless, and it has a high polarity, so
it is easy to dissolve resin compounds, fats, oils, fatty acids, carbohydrates
and other organic compounds
The extract
obtained is concentrated with a water bath until a thick extract is obtained,
which is greenish brown in bay leaves and reddish brown in secang wood. Then,
the yield calculation is carried out so that the average percent yield is
obtained, namely for bay leaves at 9.03% and the average percent yield of
secang wood is 8.44%, it can be seen in Table 4 that the percentage of yield
indicates the maximization of the solvent used to extract. The higher yield
value produced indicates the value of the extract produced more and more
The method of
determining the inhibition zone used in this study is the Kirby-Bauer disc
diffusion method. This method uses filter paper discs containing antibiotics
with certain levels placed on agar plates planted with bacteria to be examined
and then incubated. There is a zone of inhibition around the antibiotic disk,
so the bacteria examined are sensitive to the antibiotic.
From the
results of direct observation, the inhibition test of ethanol extract from bay
leaves and ethanol extract from secang wood showed a different inhibition zone,
namely the inhibition zone of secang wood is greater than bay leaves, but both
can inhibit the growth of Escherichia coli bacteria.
Statistical
tests showed that the data were normally distributed with parametric tests,
namely ANOVA Test p = 0.00, which means that there was no significant
difference in the diameter of the inhibition zone between bay leaf ethanol
extract and secang wood ethanol extract. Bay leaf ethanol extract is able to
inhibit Escherichia coli bacteria at a concentration of 60%, while secang wood
ethanol extract at a concentration of 30% is able to inhibit Escherichia coli
bacteria.
So far, people
only know diarrhea capsules derived from guava leaves. This research can be
developed into a diarrhea medicine capsule preparation derived from ethanol
extracts of bay leaves and secang wood made into dry granules as in the
original research.
CONCLUSION
The diameter
of the inhibition zone of 96% ethanol extract of bay leaves at a concentration
of 30% was 0 mm; 60% was 7.60 mm, and at a concentration of 90% was 9.60 mm. The
diameter of the inhibition zone of 96% ethanol extract of secang wood at 30%
concentration was 10.00 mm; 60% was 11.30 mm, and at 90% concentration was
14.00 mm. There is no significant difference in the effectiveness of 96%
ethanol extract of bay leaves and 96% ethanol extract of secang wood against
Escherichia coli bacteria based on the diameter of the inhibition zone.
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