Article

Antibacterial Activity of Combination of Betel Leaf Extract and Star Fruit Using

Hydroextraction Method

Dara Pranidya Tilarso^1*, Afidatul Muadifah², Windu Handaru³, Putri Indah Pratiwi⁴, Mursyidah

Lathifatul Khusna⁵, Indra Lasmana Tarigan⁶

^1,2,3,4,5Department of Pharmacy, STIKES Karya Putra Bangsa, Tulungagung Indonesia

⁶Department of Chemistry, Faculty of Science and Technology, Universitas Jambi, Indonesia

Abstract

Antibacterial compounds are bioactive substances capable of inhibiting bacterial growth by disrupting the

metabolism and cellular processes of pathogenic microorganisms. Natural plant-derived compounds have been

widely explored for their antibacterial properties, with star fruit (Averrhoa bilimbi L.) and betel leaf (Piper betle)

recognized for their potent antimicrobial effects. This study aimed to evaluate the antibacterial activity of a

combination of star fruit and betel leaf extracts against Escherichia coli and Staphylococcus aureus, two clinically

significant bacterial pathogens. The extraction process was performed using hydro-extraction at different

temperatures (40°C, 50°C, 60°C, and 90°C) to determine the optimal conditions for bioactive compound yield.

Antibacterial activity was assessed using the disc diffusion method, measuring inhibition zones to indicate of

bacterial susceptibility. The results demonstrated that the optimal inhibitory effect occurred at 50°C, producing an

inhibition zone of 19.75 mm for Staphylococcus aureus and 11.75 mm for Escherichia coli. These findings suggest that

temperature plays a critical role in maximizing the antibacterial potential of plant extracts. The study highlights the

potential application of star fruit and betel leaf extracts as natural antibacterial agents, particularly against Gram-

positive and Gram-negative bacteria. Further research is recommended to explore the mechanism of action,

phytochemical composition, and potential synergy of these extracts in antimicrobial formulations.

Keywords: Averrhoa bilimbi L; Hydroextraction, Piper betle.

Graphical Abstract

Piper betle

Averrhoa bilimbi L.

Hydroextraction Method

*

Corresponding author

Email addresses: dptilarso@stikes-kartrasa.ac.id

DOI: https://doi.org/10.22437/chp.v6i4.21736

(https://creativecommons.org/licenses/by/4.0 )

168

Chempublish Journal, 6(4) 2022, 168-176

Introduction

Antibacterial substances are compounds that

hinder bacterial growth by interfering with the

The combination of extracts allows for synergistic

activity between compounds.

The extraction technique is the process of

transferring a substance or solute from the

original solution or solid into a certain solvent [9].

Extraction techniques by maceration, soxhlet,

and hydrodistillation can produce very effective

antibacterials but require very expensive

solvents. Therefore, the hydro extraction method

was chosen using hot water by boiling and

steaming because this method is considered

more economical. Based on this, research is

needed to compare the antibacterial activity of a

combination of green betel leaf extract and

starfruit extract using the hydro-extraction

technique. The resulting extract is expected to be

more effective and cost-efficient as an antiseptic

in inhibiting the growth of Staphylococcus aureus

and Escherichia coli.

metabolism

of

harmful

microbes

[1].

Antibacterial substances derived from synthetic

materials can prevent bacterial infections, but

many have side effects such as irritation. This

problem encourages the shift in the use of

antibacterial substances from synthetic materials

to natural materials. Natural material extracts

with antibacterial content can be formulated into

antiseptic preparations through hand sanitizers

or hand soap. Plants with antibacterial properties

include starfruit (Averrhoa bilimbi L.) and betel

leaves (Piper betle). These plants are useful as

traditional medicines that our ancestors have

long used. Starfruit is effective in treating various

conditions,

diabetes,

including

coughs,

rheumatism, canker sores, mumps, toothaches,

acne, bleeding gums, diarrhea, and high blood

pressure [2]. Starfruit contains compounds such

as saponins, flavonoids, steroids/triterpenoids,

Material and Methods

Materials and Instrumentations

and

tannins.

These

compounds

exhibit

antibacterial activity by inhibiting protein

synthesis [3]. According to a study by Aifianti

(2014), revealed that starfruit extract displays

antibacterial activity against Staphylococcus

aureus and Escherichia coli bacteria at a 10%

concentration. An extract concentration of 10%

or the lowest concentration can inhibit bacterial

growth and produce an inhibition zone diameter

[4]. Meanwhile, betel leaves are efficacious for

treating vaginal discharge, eliminating bad

breath, treating wounds, stopping bleeding

gums, mouth ulcers, and eliminating body odor

[5]. The chemical content found in betel plants is

saponin, flavonoids, polyphenols, and essential

oils [6]. Saponin compounds exhibit antibacterial

effects by damaging the cytoplasmic membrane,

Piper betle and Averrhoa bilimbi extracts, test

bacteria Staphylococcus aureus and Escherichia

coli. The test media including Nutrient Agar (NA)

and Nutrient Broth (NB). Additional materials are

distilled

Dragendorff's

water

(H₂O),

reagent,

Mayer's

reagent,

concentrated

hydrochloric acid (HCl P), Iron (III) Chloride (FeCl₃),

ethyl acetate (C₄H₈O₂), anhydrous acetic acid,

concentrated sulfuric acid (H₂SO₄), hot water,

hydrochloric acid (HCl). Other tools are knives,

trays, cloths, stoves, pans, beaker glasses,

thermometers, petri dishes, measuring cups,

droppers, test tubes, test tube racks, analytical

scales, spirit lamps, dropper plates, erlenmeyer

flasks

which

results

in

cell

death.

Flavonoid

Methods

compounds, meanwhile, work by causing the

denaturation of bacterial cell proteins [7]. Betle

leaf extract at a 40% concentration can inhibit

Staphylococcus aureus bacteria, producing an

inhibition zone with a diameter of 17.33 mm [8].

Based on the content of compounds in the two

plants that have antibacterial activity, both plants

have the potential to be used as natural

antiseptics. The combination of these two

extracts is anticipated to offer enhanced

effectiveness in suppressing bacterial growth.

Sample Preparation and Extraction. The plant

materials used in this study consisted of green

betle leaves (Piper betle) and starfruit (Averrhoa

bilimbi L.), sourced from Bendiljati Wetan Village,

Tulungagung, and identified at Materia Medica

Batu, Malang. For extract preparation, several

fresh yellow starfruits and green betel leaves

were selected, washed with clean water, and then

allowed to dry at room temperature. Once dried,

the samples were cut into small pieces of

169

Chempublish Journal, 6(4) 2022, 168-176

approximately 0.5 cm and weighed a total of 100

g. The hydro extraction method was employed,

which involved boiling at temperatures of 40°C,

50°C, and 60°C, as well as steaming at 90°C [10].

For the boiling method, 100 ml of water is

prepared and heated in a water bath, with

temperature settings of 40°C, 50°C, and 60°C.

Then, 100 g of the cut samples are added to the

heated water and left for 30 min. Afterwards, the

mixture is filtered using plastic gauze for the first

filtration, followed by a second filtration using

filter paper. Treatment with the steaming

method, prepare enough water in a steamer that

has been perforated on the cover and insert a

thermometer in the perforated part for

temperature control, insert 100 g of sample, heat

on the stove at a temperature of 90ºC, wait

approximately 30 min until the extract is

obtained then filtered with filter paper [11].

Extract

Analysis

using

Liquid

Chromatography – High Resolution Mass

Spectrometry (LC-HRMS). The combination

extract of green betel leaf (Piper betle) and

starfruit (Averrhoa bilimbi L.) was analyzed using

a Liquid Chromatography system. The solvents

used were A = 0.1% Formic Acid in Water and B =

0.1% Formic Acid in Acetonitrile. The analytical

column employed was Hypersil GOLD aQ, with

dimensions of 50 x 1 mm and a particle size of 1.9

µm. The analytical flow rate was set at 40 µL/min,

and the flow gradient followed the pattern shown

in Table 1. The operating time was 30 min, with

the column oven temperature maintained at

30°C. HRMS analysis was conducted with a full

scan at a resolution of 70,000, with MS2 data

captured at a resolution of 17,500. The total

running time was 30 min, and the analysis was

performed in both positive and/or negative

polarity.

Phytochemical screening. Flavonoids: One gram

of the sample extract is placed in a test tube,

followed by the addition of concentrated HCl and

heating in a water bath for 15 minu. The

formation of a red or yellow color indicates a

positive result for flavonoids, including flavones,

chalcones, and aurones [12]. Tannins: Two grams

of the extract are dissolved in ethanol until fully

submerged. Subsequently, 1 mL of the solution is

transferred into a test tube, and 2–3 drops of 1%

FeCl₃ solution are added. The presence of

tannins is confirmed by the appearance of a

bluish-black or green color [13]. Alkaloids: The

Table 1. Gradient Elution

No Time

Flow Rate%B

(µL/min)

Curve

1

2

3

4

5

6

7

8

9

0.000

Equilibration

5.0

0.000

40.000

5

New Row

0.000

Run

2.000

40.000

5.0

5

15.000

22.000

25.000

25.100

60.0

95.0

5.0

identification

conducted

of

using

alkaloid

Mayer's

compounds

reagent

is

and

Dragendorff's reagent. Terpenoids: Two grams of

the sample extract are introduced into a test

tube, followed by the addition of 2 mL of ethyl

acetate, then shaken. The ethyl acetate layer is

separated, transferred onto a dropper plate, and

left to dry. Once dried, 2 drops of anhydrous

acetic acid and 1 drop of concentrated sulfuric

acid are added. The formation of a red or yellow

color indicates a positive result for terpenoids,

whereas a green color signifies the presence of

steroids [12]. Saponin: Half a gram of the

extracted sample is mixed with 0.5 mL of hot

water and shaken for 1 minute. If foam appears,

1N HCl is added. The extract is considered

positive for saponins if the foam remains stable

for 10 min at a height of 1–3 cm. [14].

10 30.000

11 New Row

12 30.000

5.0

Stop Run

Antibacterial Activity. The antibacterial test

begins with the preparation of nutrient agar

media. To prepare the media, 2 g of agar powder

are weighed and dissolved in 100 mL of distilled

water. The solution is stirred until fully

homogeneous and then sterilized in an autoclave

at 121°C for 15 min. After sterilization, extract

comparisons between starfruit and green betle

leaves are made at a ratio of 20:10 mg/mL, with

170

Chempublish Journal, 6(4) 2022, 168-176

chloramphenicol serving as the positive control

and distilled water as the negative control. Next,

S. aureus and E. coli bacteria are evenly inoculated

on each agar plate. After inoculation, 10 μL of the

extract and 10 μL of antibiotics are applied to the

designated disc paper on the plates. After the

installation of the disc paper is complete, the

bacteria are incubated at 37°C for 24 hr, then the

diameter of the inhibition zone is observed and

measured. The diameter of the inhibition zone is

measured using a calliper, and the results are

then compared with the positive controls,

negative controls, and classification values of the

inhibition zone diameter.

(Table 1). These findings suggest that the

combination may possess potential medicinal

and antioxidant properties, whereas saponins,

steroids, and terpenoids were not detected.

Profiling

Liquid

Chromatography

–

High

Resolution Mass Spectrometry (LC-HRMS)

Liquid Chromatography-High Resolution Mass

Spectrometry (LC-HRMS) analysis revealed that

the combination extract of green betel leaf (Piper

betle L.) and star fruit (Averrhoa bilimbi L.)

contains bioactive compounds with potential

antibacterial properties (Table 3). Among the

identified compounds, apigenin, a flavonoid, was

detected at a retention time of 0.870 min, with a

molecular weight of 271.05991 g/mol (Table 3).

Results and Discussions

The sample was extracted using distilled water

because distilled water is neutral, has no

antibacterial activity, is not easily evaporated and

is polar [15]. The method used is the

hydroextraction method because this method is

more effective and efficient so that it can save

production costs. In addition, this method is also

easier to apply by the community [10].

Flavonoids, including apigenin, are known for

their antibacterial activity, primarily through

interactions with extracellular and soluble

proteins, leading to bacterial cell membrane

disruption

intracellular

and

components

subsequent

[16].

leakage

Specifically,

of

apigenin has been reported to inhibit the growth

of Staphylococcus aureus and Escherichia coli, two

Table 2. Secondary metabolites of Combination

Extract

clinically significant bacterial

[16-17]. The

antibacterial mechanism of apigenin is linked to

its ability to inhibit the glucosyltransferase

enzyme, which plays a critical role in bacterial

Secondary Metabolite

Result

Alkaloids

Flavonoids

Tannins

+

-

adhesion

and

biofilm

formation,

thereby

reducing bacterial colonization and persistence

[18].

Saponins

Steroids

-

These findings align with previous studies

highlighting flavonoids as promising natural

antibacterial agents that could serve as

alternatives or complementary treatments to

Terpenoids

-

Note: (+) contains coumpounds; (-) does not contain

compounds

The

phytochemical

screening

test

of

a

conventional

antibiotics,

particularly

in

combination of green betel leaf extract and star

fruit in a 2:1 ratio was conducted by observing

the color changes produced by phytochemical

reagents in the test tube, which indicated the

presence of specific compounds in the extract.

The results showed that the combination

contained flavonoids, alkaloids, and tannins

addressing antibiotic-resistant bacterial strains

[19]. The presence of apigenin in the combination

extract of Piper betle L. and Averrhoa bilimbi L.

further underscores the therapeutic potential of

plant-based

bioactive

compounds

in

the

development of novel antibacterial formulations.

Table 3. Antibacterial Active Compounds in Combination Extracts

171

Chempublish Journal, 6(4) 2022, 168-176

Molecular

Weight

(g/mol)

Retention

Time (min)

Name

Formula

Area (max)

m/z

Apigenin

Piperine

Betaine

C₁₅H₁₀O₅

C₁₇H₁₉NO₃

C₅H₁₁NO₂

271.05991

286.14337

118.0863

0.870

35,328,318.48

4,570,066,672.75

101,427,056.68

99.2

98.7

98.2

14.602

20.442

Figure 1. Total Ion Chromatogram

surface of Staphylococcus aureus bacterial cells

[20].

Figure 2. Single chromatogram of Apigenin from

LC analysis

Figure 4. Single chromatogram of Piperine from

LC analysis

Figure 3, Apigenin Mass Spectrum

Figure 5. Piperine Mass Spectrum

The antibacterial mechanism of piperine against

Staphylococcus aureus is as a protein A inhibitor.

Protein A is a protein that is only found on the

172

Chempublish Journal, 6(4) 2022, 168-176

Betaine compound in the combination extract of

betel leaf and starfruit can be eluted at a

retention time of 20.442 min (Figure 6) with a

molecular weight of 118.0863 (Figure 6). Betaine

compound is included in the Tannin compound

group. The antibacterial mechanism of tannin

has an antibacterial effect by implementing

inhibition of protein synthesis. The antibacterial

effect of tannins occurs through interactions with

cell membranes, enzyme inactivation, and the

disruption of genetic material. The mechanism

by which tannins act as antibacterials involves

inhibiting the reverse transcription enzyme and

DNA topoisomerase, preventing bacterial cell

formation [16].

Antibacterial Test of Combination of Green Betel

Leaf Extract and Starfruit Fruit

The antibacterial activity of the combined green

betle leaf and starfruit extracts was evaluated

using the disc diffusion method. The test plates

were incubated at 37°C for 24 hr. To assess the

optimum inhibitory effect, the diameter of the

clear zone surrounding the disc was measured

using a caliper. The results, illustrating the

formation of inhibition zones around the disc due

to the extract combination, are shown in Figure 8

A

B

Figure 6. Single chromatogram of Betaine from

Figure 8. Inhibition of combination of betle leaf

extract and starfruit. (A) Staphylococcus aureus,

(B) Escherichia coli.

LC analysis

Figure 7. Betaine Mass Spectrum

Table 4. Diameter of inhibition zone against Staphylococcus aureus and Escherichia coli

Diameter Clear Zone (mm)

Treatment

S. aureus

R3

E. coli

R1

R2

R1

R2

R3

Mean

40°C

50°C

60°C

90°C

K+

5

24.5

14.75

8

10

9

17

22.5

19.5

20.5

19.75

16.25

14.5

35.5

0

19.75

16.25

14.4

29.5

0

13

15.5

8

9

8

11

13

11.75

8

11.75

8

8.5

27.5 25.5

33

0

5.5 28

22.17

0

K-

0

R= replication; K+= Positive control (Kloramfenikol); K- = Negative control (aquades)

The results of the antibacterial activity test of the

combined extract demonstrated optimal effects

at 50°C, producing an inhibition zone of 19.75

mm against Staphylococcus aureus, indicating a

strong antibacterial response. Additionally, at

60°C, the extract exhibited optimal activity

against Escherichia coli, forming an inhibition

173

Chempublish Journal, 6(4) 2022, 168-176

zone of 11.75 mm, which signifies a moderate

inhibitory effect on bacterial growth. (Table 4).

alkaloid- and tannin-rich plant extracts continue

to be explored as potential alternatives or

adjuvants

to

conventional

antibacterial

The phytochemical screening of the starfruit and

betel leaf extract combination confirmed the

presence of alkaloids and tannins. Alkaloids are

bioactive compounds that act as antibacterial

compounds just like bioactive phenol, flavonoid,

and tannin compounds. The mechanism is by

damaging cell metabolism so that bacterial

growth is inhibited.

treatments [23-24]. Tannins interact with

polypeptides in the bacterial cell wall, disrupting

its proper formation. This structural instability

makes bacteria susceptible to lysis due to

osmotic or physical pressure, ultimately leading

to cell death [4].

The study conducted indicates the combination

of starfruit extract and betel leaf effectively

inhibits the growth of Staphylococcus aureus in

the strong category and Escherichia coli in the

moderate category. However, the inhibition zone

diameter for both bacteria remains smaller than

that of the positive control. Staphylococcus

aureus is classified as a gram-positive bacterium,

whereas Escherichia coli is gram negative. The

cell wall of gram-positive bacteria contains a

higher concentration of peptidoglycan and fewer

lipids, along with polysaccharides. Peptidoglycan

is composed of amino acids and sugars, while

teichoic acid, a water-soluble polymer, facilitates

the transport of positive ions. The polar nature of

the gram-positive bacterial cell wall suggests that

it is more permeable to polar compounds. Since

alkaloids and tannins are polar, they can more

readily penetrate the polar peptidoglycan layer

compared to the non-polar lipid layer found in

gram negative bacteria. This leads to a stronger

inhibitory effect on gram positive bacteria than

on gram negative bacteria [22].

Natural plant-derived compounds, particularly

alkaloids and tannins, have been widely studied

for

Alkaloids

their

potent

exert their

antibacterial

properties.

effects

antibacterial

primarily by disrupting the peptidoglycan

structure of bacterial cell walls, an essential

component for maintaining cell integrity and

shape.

By

interfering

with

peptidoglycan

synthesis, alkaloids hinder proper cell wall

formation, leading to increased cell permeability,

osmotic instability, and ultimately bacterial cell

death. In addition to targeting the cell wall, some

alkaloids have been reported to interfere with

bacterial nucleic acid synthesis and inhibit key

metabolic enzymes, further contributing to their

bactericidal activity [20].

Similarly, tannins play a significant role in

antibacterial defense by inhibiting essential

bacterial enzymes, such as reverse transcriptase

and DNA topoisomerase, which are critical for

bacterial replication and transcription [21]. By

blocking DNA synthesis and replication, tannins

Conclusion

effectively

prevent

bacterial

growth

and

proliferation. Moreover, tannins interact with

bacterial cell membranes, leading to protein

The analysis of active compounds in the

combination extract confirmed the presence of

alkaloids and tannins. Antibacterial testing

precipitation,

enzyme

inactivation,

and

disruption of nutrient transport systems, all of

which contribute to bacterial cell death [22].

demonstrated

exhibited optimal inhibition at 50°C, with an

inhibition zone measuring 19.75 mm, indicating a

that

Staphylococcus

aureus

These antibacterial mechanisms highlight the

therapeutic potential of alkaloids and tannins as

natural antimicrobial agents, particularly in the

fight against antibiotic-resistant bacteria. Recent

studies suggest that combining alkaloids and

tannins with conventional antibiotics may

strong

antibacterial

effect.

Meanwhile,

Escherichia coli showed peak activity at 60°C,

forming an inhibition zone of 11.75 mm, which

suggests a moderate antibacterial response.

Further studies are recommended to explore the

antibacterial

combination through in vivo testing.

properties

of

this

extract

enhance antimicrobial efficacy, providing

a

promising strategy to combat multidrug-

resistant bacterial infections. As research in

phytochemistry and pharmacology advances,

174

Chempublish Journal, 6(4) 2022, 168-176

Bakteri Staphylococcus aureus secara In

Acknowledgement

vitro.

Jurnal

MIPA,

2(2),

128.

Thank you to the Directorate of Research and

Community Service (DRPM) for funding this

research with contract number 159 / E5 /

https://doi.org/10.35799/jm.2.2.2013.3121

Dewi, L.K., Sarosa, A.H., Kartikowati, C.W.,

Hayati, N., Parasu, R., Amalia, E. (2021).

Pengaruh Jenis Pelarut Terhadap Daya

Antibakteri Hasil Ekstraksi Daun Sirih Hijau

(Piper betle L.) pada Aktivitas Staphylococcus

Epidermidis. Journal of Innovation and

[5]

P6.02.00.PT

/

2022

from

the

APBN

Implementation List (DIPA) of the Deputy for

Strengthening Research and Development of the

Ministry of Research and Technology/National

Research and Innovation Agency in 2022.

Applied

Technology.

Vol.

7

(1).

OI: http://dx.doi.org/10.21776/ub.jiat.2021

.007.01.6

Author Contributions

[6]

[7]

[8]

Noventi, W. R.-4272-2-P. pdfa., & Carolia, N.

(2016). Potensi Ekstrak Daun Sirih Hijau (

Piper betle L .) sebagai Alternatif Terapi

Acne vulgaris. Studi Pendidikan Dokter

Fakultas Kedokteran Universitas Lampung,

Vol. 5(1), Hal. 140.

Conceptualization, DPT and AM.; Methodology,

WH, PIP, and MLK; Software, DPT, WH, and AM.;

Validation: DPT and AM; Formal Analysis, DPT

and AM.; Investigation, WH, PIP, and MLK;

Resources, DPT and AM.; Data Curation, AM and

AM; Writing – Original Draft Preparation, DPT,

AM, and ILT; Writing – Review & Editing, DPT, AM

and ILT and ML; Visualization: DPT and AM.;

Supervision, DPT and AM; Project Administration,

DPT.

Triyani, M. A., Pengestuti, D., Khotijah, S. L.,

Susilaningrum, D. F., & Ujilestari, T. (2021).

Aktivitas

Antibakteri

Hand

Sanitizer

Berbahan Ekstrak Daun Sirih dan Ekstrak

Jeruk Nipis. NECTAR: Jurnal Pendidikan

Biologi, 2(1), 16-23

Bagus, I., Suyasa, O., Bekti, H. S., Rinawati,

L. P., & Laksmi, L. P. (2022). Daya Hambat

Ekstrak Daun Sirih dan Daun Legundi

Conflic of Interest

There are no significant conflicts that interfere

with the progress of this research, from data

collection to processing results and conclusions.

Terhadap

Pertumbuhan

Bakteri

Staphylococcus aureus. 1(5). The Journal Of

Muhammadiyah

Technologist.

Medical

Laboratory

References

[1]

[2]

[3]

[4]

Rustanti, E., Jannah, A., & Fasya, A. G.

(2013). Uji Aktivitas Antibakteri Senyawa

Katekin Dari Daun Teh (Cameliasinensis

L.Var

Micrococcusluteus.

[9]

Aji, A., & Bahri, S. (2017). Pengaruh Waktu

Ekstraksi Dan Konsentrasi HCl untuk

Pembuatan Pektin dari Kulit Jeruk Bali

(Citrus maxima). Jurnal Teknologi Kimia

Unimal, 6(1), 33–44.

Assamica)

Terhadap

Bakteri

Alchemy,

2(2).

https://doi.org/10.18860/al.v0i0.2886

[10] Rahayu, N. W. S., Prasetyo, E. N., &

Isdiantoni. (2016). Hindroekstraksi Daun

Ketapang (Terminalia catappa L.) sebagai

Pengendali Penyakit Ice-ice pada Budidaya

Kappaphycus alvarezii. Jurnal Sains Dan

Seni Its, 1–8.

Maryam, S., Juniasti, S., & Kosman, R.

(2015). Uji Aktivitas Antibakteri Ekstrak

Etanol Buah Belimbing Wuluh (Averrhoa

Bilimbi L.) Asal Kota Watampone. Jurnal

Ilmiah

As-Syifaa,

7(1),

60–69.

https://doi.org/10.33096/jifa.v7i1.21

[11] Indrayati, A., Farmasi, F., Buana, U.,

Ferdyani, S., Yuniarto, P. F., (2020). Uji

Aktivitas Antibakteri Sediaan Gel Ekstrak

Etanol Buah Belimbing Wuluh (Averrhoa

Karawang, P.,

Formulasi

&

Pudding, S. (2019).

,

Uji Stabilitas Fisik Dan

Kompatibilitas Produk Kosmetik Anti-Aging

Dalam Sediaan Serum. Jurnal Ilmiah

Farmasi, 4, 2.

Bilimbi

Linn)

terhadap

Bakteri

Staphylococcus aureus. Jurnal Kesehatan

Mahasiswa UNIK, 2(1), 30–42.

Ngajow, M., Abidjulu, J., & Kamu, V. S.

(2013). Pengaruh Antibakteri Ekstrak Kulit

Batang Matoa (Pometia pinnata) terhadap

[11] Muthmainnah. (2017). Skrining Fitokimia

Senyawa Metabolit Sekunder Dari Ekstrak

Etanol Buah Delima (Punica Granatum L.)

175

Chempublish Journal, 6(4) 2022, 168-176

Dengan Metode Uji Warna. Media Farmasi,

13(3), 1576–1580.

[12] Harborne, J.B. (2006). Metode Fitokimia:

[20] Yan Y, Li X, Zhang C, Lv L, Gao B, Li M.

Research

Progress

on

Antibacterial

Activities and Mechanisms of Natural

Penuntun Cara Modern Menganalisis

Alkaloids: A Review. Antibiotics (Basel).

Tumbuhan

(alih

bahasa:

Kosasih

2021

Mar

19;10(3):318.

doi:

Padmawinata & Iwang Soediro). Bandung :

https://doi.org/10.3390/antibiotics100303

Penerbit ITB.

18 .

[13] Dewi, N. R. K., Kuncoro, H., & Rijai, L. (2015).

Potensi Sitotoksik Ekstrak Air Daun Sirih

Hitam (Piper sp.). Jurnal sains dan

Kesehatan, 1(1), 11-15.

[21] Vaou N, Stavropoulou E, Voidarou C,

Tsigalou C, Bezirtzoglou E. Towards

Advances in Medicinal Plant Antimicrobial

Activity: A Review Study on Challenges and

Future Perspectives. Microorganisms. 2021

[14] Dewantoro,

Z.,

Widodo,

Y.L.A.,

Ciptaningtyas,

Pemberian Ekstrak daun Belimbing Wuluh

(Averrhoa

Pertumbuhan

aureus secara In Vitro. Jurnal Kedokteran

Diponegoro, 6(2); 886-892

R.

(2017).

Pengaruh

Sep

27;9(10):2041.

doi:

https://doi.org/10.3390/microorganisms91

02041

bilimbi

Bakteri

L).

Terhadap

Staphylococcus

[22] Redondo LM, Chacana PA, Dominguez JE,

Fernandez Miyakawa ME. Perspectives in

the use of tannins as alternative to

antimicrobial growth promoter factors in

[15] Parama, P. W., Sukrama, I. D. M., &

Handoko, S. A. (2019). Uji efektifitas

antibakteri ekstrak buah jeruk nipis (Citrus

poultry.

27;5:118.

Front

Microbiol.

2014

Mar

doi:

aurantifolia)

Streptococcus mutans in vitro. Bali dental

journal, 3(1), 45-52.

terhadap

pertumbuhan

https://doi.org/10.3389/fmicb.2014.00118

[23] Sari, D. R. A. P., Yustiantara, P. S., Paramita,

N. L. P. V., & Wirasuta, I. M. A. G. (2014). Uji

Aktivitas Antibakteri Ekstrak Etanol Buah

Lada Hitam (Piper nigrum L.) Terhadap

Bakteri Propionibacterium acnes. Jurnal

Farmasi Udayana, 3(2), 279831.

[16] Nuria, M.C., A. Faizatun., dan Sumantri.

2009. Uji Antibakteri Ekstrak Etanol Daun

Jarak Pagar (Jatropha cuircas L) terhadap

Bakteri Staphylococcus aureus ATCC 25923,

Escherichia coli ATCC 25922, dan Salmonella

typhi ATCC 1408. Jurnal Ilmu –Ilmu

Pertanian. 5: 26 –37.

[24] Tarigan, I., Muadifah, A., & Latief, M. (2023).

Non-Ribosomal

Inhibition

of

The

Antibacterial Compound from Ethanol

[17] Isnarianti, R., Wahyudi, I. A., & Puspita, R. M.

(2013). Muntingia calabura L leaves extract

inhibits glucosyltransferase activity of

Streptococcus mutans. Journal of Dentistry

Indonesia, 20(3), 59-63.

Extract of Gnetii gnemon. Life Science, 12(2),

174-185.

https://doi.org/10.15294/lifesci.v12i2.6718

3

[18] Matilla-Cuenca, L., Gil, C., Cuesta, S. et al.

Antibiofilm activity of flavonoids on

staphylococcal biofilms through targeting

BAP amyloids. Sci Rep 10, 18968 (2020).

https://doi.org/10.1038/s41598-020-

75929-2

[19] Shamsudin NF, Ahmed QU, Mahmood S, Ali

Shah SA, Khatib A, Mukhtar S, Alsharif MA,

Parveen H, Zakaria ZA. Antibacterial Effects

of Flavonoids and Their Structure-Activity

Relationship

Interpretation.

9;27(4):1149.

Study:

Molecules.

A

Comparative

2022

Feb

doi:

https://doi.org/10.3390/molecules270411

49

176