Combination Test Chinese Leaves Extract (Leucaena leucocephala
folium) and Aloe Vera Inhibiting Growth Escherichia coli
Putra Rahmadea Utami1, Chairani1, Hendra Yudha1
1

Department of Medical Laboratory
Technology, Fakultas of Health
Science, Universitas Perintis
Indonesia, Padang, Indonesia
Correspondence:
Putra Rahmadea Utami,
Universitas Perintis Indonesia,
Jl. Adinegoro KM 17, Simpang
Kalumpang, Lubuk Buaya, Padang,
West Sumatera, Indonesia
Zip Code : 25586
Email:
putrarahmadeautami123@gmail.com
Received: June 10, 2020
Revised: Juni 29, 2020
Accepted: August 20, 2020

Abstract
Chinese petai (Leucaena leucocephala folium) and aloe vera
(Aloe vera L.) have medicinal properties among the
plants. The objective of this study was to determine the
number of inhibitory zones produced by the ethanol extract
of Chinese petai and aloe vera on the growth of Escherichia
coli. The research method was In-vitro Experimental
Laboratory research design with the Kirby Bauer method.
The samples used was Chinese petai and aloe vera with pure
strains of E. coli. One Way ANOVA was used to compare
the differences in inhibition of Chinese petai and aloe vera
on the growth of E. coli. The results of a combination of
Chinese petai and aloe vera extract test showed that there
were significant differences in the concentration of 25 g, 50
g, 75 g, and 100 g (p <0.05). The ethanol extract of Chinese
petai and aloe vera can inhibit the growth of E. coli. From
the results of this study found that there was an interaction
on the combination of ethanol extract of Chinese petai and
aloe vera inhibiting the growth of E. coli with the most
effective concentration being 100 g/mL. This study can find
out the benefits of petai cina and aloe vera also the public
will know the benefits and efficacy of Chinese petai and aloe
vera leaves in medicine.
Keywords
Leucaena leucocephala folium, Aloe vera L., Escherichia
coli, inhibition, combination

calcium, phosphorus, iron, fatty acids, fibre,

INTRODUCTION
(Leucaena

vitamin A and other vitamins. Besides that,

leucocephala folium) and aloe vera (Aloe

Chinese petai leaves also contain various

vera) are one of the traditional medicines that

flavonoids that become anti-inflammatory

have the potential to be developed and

and

utilized by the people of Indonesia (1).

stimulate skin cell growth while the alkaloids

Chinese petai contains alkaloids, saponins,

function

lectins, tannins, mimosin, leucanin, protein,

beneficial properties, including tannin, amino

Chinese

petai

leaf

antioxidants.

as

Lectins-contained

antimicrobials.

Aloe

to

vera

60

Putra Rahmadea Utami, et al.

acids, anthraquinone which is a phenolic

closed seed plants, can grow in dry areas and

compound

also in cold climates (7,8).

that

acts

as

a

laxative,

antimicrobial agent and has a strong

E.

coli

belongs

to

the

family

analgesic effect. It also has campesterol,

Enterobacteriaceae. This bacterium is a

sitosterol and lupeol compounds. These

Gram-negative bacterium, in the form of a

compounds act as an anti-inflammatory and

short stem, has a flagellum, measuring 0.4 to

antibacterial agent (1,2).

0.7 μm x 1.4 μm. E. coli grows well in almost

Escherichia coli is the bacteria that most

all breeding media can ferment lactose and is

often cause urinary tract infections and

micro-aerophilic. These bacteria become

nosocomial infections. E. coli can cause

pathogens when outside intestinal tissue.

primary intestinal infection and also used to

Clinically frequently infected sites are the

assess whether a water supply for household

urinary tract, bile duct, and other places in the

use is good or not.

E. coli is a bacteria

abdominal cavity and any anatomic site (e.g.

commensal which can pathogenic, acting as a

prostate gland, lung, bone) can be the site of

primary cause of morbidity and mortality

disease (9). E. coli is a gut flora in the

throughout the world (1,3,4).

digestive tract that can turn into pathogenic

Chinese lamtoro and petai plants come

opportunists and cause diseases such as

from tropical America that has a deep root

diarrhoea. This bacterium has various

system and resistant to dry conditions.

pathogenic

Chinese petai contains alkaloids, saponins,

Enteropathogenic Escherichia coli (EPEC),

flavonoids, lectins, calcium, phosphorus,

Enterotoxigenic Escherichia coli (ETEC),

iron, tannins, mimosin, leukanin, proteins

Enteroinvansive Escherichia coli (EIEC) and

vitamin A and vitamin B (6). The leaves

Enterohemorrhagic Escherichia coli (EHEC)

contain 22% protein (4). Seeds, leaves and all

(10,11).

parts of the plant can use to treat several

strains,

including

Combination therapy uses to broaden the

diseases, such as diabetes, fracture, intestinal

spectrum

worms,

menstruation,

toxicity, slow down the process of antibiotic

inflammation of the kidneys (nephritis) and

resistance and reduce the dose of the drug and

insomnia.

has

ulcer,

late

of

antimicrobials,

considerable

antimicrobial

minimize

activity.

Aloe vera is a traditional medicinal plant

Recently, there has been no information

that has many benefits and benefits. It is a

about the effective concentration of the

cactus-like plant originating from Africa,

combination of extracts of Chinese petai

which belongs to the Liliaceae family and

leaves and aloe inhibiting the growth of the

Ina J Med Lab Sci Tech 2020; 2(2): 60-67

61

bacteria E. coli. The purpose of this study

Putra Rahmadea Utami, et al.

was to analyze the interaction effects of

physiological NaCl (in packaging), steril

ethanol extracts of Chinese petai leaves and

empty disks, Aquades as negative controls

aloe vera leaves in inhibiting the growth of E.

and antibiotic as the positive control. While,

coli (4,5).

the culture media used were MHA (Muller
Hilton Agar), NB (Nutrient Broth), Endo

MATERIALS AND METHODS
This research conducted using the

Solution.

experimental

The extraction method used to extract

laboratory method. The research method

Chinese petai plants and aloe vera is a cold

involves making ethanol

method

completely

randomized

extract

from

extraction

method,

namely

Simplicia (Crude drugs) by maceration using

maceration. Chinese petai leaves that turned

96% ethanol solvent. It also examines the

into Simplicia powder are added with 70%

interaction between ethanol extracts of

ethanol solvent until submerged. Maceration

Chinese petai leaves and aloe vera stem on

poured and squeezed. The pulp is macerated

the inhibitory power of E. coli and in vitro by

again with a new solution until submerged.

measuring the inhibitory by diffusion method

The maceration process carried out three

according to Kirby Bauer. The principle of

times then it was evaporated on a rotary

the Kirby Bauer method is the inhibition of

evaporator until a thick extract of Chinese

the growth of microorganisms, the diameter

petai leaf obtained then lastly weighed to

of the zone of inhibition of bacterial growth

determine its weight.

shows the sensitivity of the bacteria, the

Extraction of Chinese Petai Leaves and

wider the zone diameter formed by bacteria,

Aloe Vera

the better it is to inhibit bacterial growth.

62

Agar, Nutrient Agar, and Mc Farland

Aloe vera leaves peeled first to get as

The tools used in this study were Petri

much as 1000 grams of aloe vera gel mashed

dishes, tissue cotton, test tubes, wire mesh,

with a blender then soaked with 96% ethanol

syringes, matches, bunsen burner analytical

solvent until aloe vera is submerged

scales, slide callipers, microscopes, sterile

completely, after that it is allowed to stand for

stick cotton, Erlenmeyer, beaker glass,

2-3 days in a closed jar. Then, the liquid

parchment

ovens,

extract was filtered with gauze filter and

incubators, label paper, measuring pipettes,

collected the extract in a bottle. Steering of all

water baths, paper cutters, rotary evaporators.

filtrates obtained from the immersion.

The materials used in this study are pure

Concentrated with a rotary evaporator until

strains of E. coli ethanol extracts of Chinese

thick extract is obtained (1).

petai

leaves

paper,

and

tweezers,

aloe

vera,

sterile
Ina J Med Lab Sci Tech 2020; 2(2): 60-67

Putra Rahmadea Utami, et al.

Extract Determination
The

extract

yield

RESULTS
calculated

by

Inhibition test of the ethanol extract of

comparing the weight of the ethanol extract

Chinese Petai Leaves and Aloe Vera with the

obtained with the initial sample weight. The

disc diffusion method that carried out with

yield obtains through this formula: (extract

various concentrations of the extract.

weight obtained/initial sample weight) x
100%.

Table 1. Results of Inhibitory Tests of Chinese Petai Leaf Extract Against Escherichia coli
Extract Concentration
(mg/mL)
25
50
75
100
Control (+) Chloramphenicol

Extract
Repetition (mm)
1
2
3
0
0
0
0
0
0
6
6
7
7
8
8
30
29
30

X̄

SD

0
0
6.3
7.7
29.7

± 0.
± 0.
± 0.58
± 0.58
± 0.58

P. sig

0.00

Table 2. Results of Inhibitory Aloe Vera Leaf Extract Against Escherichia coli Bacteria (mm)
Extract Concentration
(mg/mL)
25
50
75
100
Control (+) Chloramphenicol

Extract
Repetition (mm)
1
2
3
8
10
9
10
11
10
12
12
11
14
15
14
30
28.5
30

X̄

SD

9
10.3
11.7
14.3
29.5

1
0.58
0.58
0.58
0.87

P. Sig

0.00

Table 3. Test Results of Combination Inhibition of Chinese Petai Leaf Extract and Aloe Vera
on Escherichia coli (mm)
Extract Concentration
(mg / ml)

Control (+) Chloramphenicol

Ina J Med Lab Sci Tech 2020; 2(2): 60-67

X̄

SD

6.3
8.3
10.3
11.7
27.2

0.58
0.58
0.58
0.58
1.26

P. Sig

0.00

63

25
50
75
100

Extract
Repetition (mm)
1
2
3
6
7
6
8
9
8
10
11
10
12
12
11
27
28.5
26

Putra Rahmadea Utami, et al.

Fig 1. The results of the test for the inhibition of aloe vera against Escherichia coli with a
concentration of 4 samples. Each sample size was carried out 3 times. In the middle is the (+)
chloramphenicol control

Based on Table 1, the inhibitory test

it increases following the increase of the

results of Chinese Petai leaf extract against E.

extract concentrations. A positive control

coli (mm) bacteria with a concentration of 75

using

mg/mL formed inhibition zone 6.3 ± 0.58

obtained the broadest inhibition zone (29.5 ±

mm. Moreover, the amount of 100 mg/mL

0.87 mm) compared to others.

extract created an inhibition zone of 7.7 ±
0.58

mm.

A

positive

control

Based

the Chloramphenicol antibiotic

on

Table

3,

the

extract

using

concentration of 25 mg/ml formed an

the Chloramphenicol antibiotic obtained the

inhibition zone 6.3 ± 0.58 mm, its increases

broadest inhibition zone (29.7 ± 0.58 mm)

following the increase of the extract

compared to other concentrations.

concentrations. A positive control using

Based on Table 2, the inhibitory test

the Chloramphenicol antibiotic obtained the

results of Aloe Vera extract against E.

broadest inhibition zone (27.2 ± 1.26 mm)

coli (mm) bacteria with a concentration of 25

compared to others.

mg/mL formed inhibition zones 9± 1 mm and

DISCUSSION

64

Interaction of the combination of ethanol

close together. The test to identify the
synergistic

effect

of

antibacterial

extract of Chinese petai leaves and aloe vera

combinations, discs used and treated with a

against E. coli bacteria showed in the test of

single antimicrobial agent each, then both are

direct mixing of the two antibacterial agents.

placed at a distance equal to the average

It formed the inhibitory zone when placed

number of the radius of the zone of inhibition
Ina J Med Lab Sci Tech 2020; 2(2): 60-67

Putra Rahmadea Utami, et al.

of the antimicrobial agent when tested

cytoplasmic membrane in bacteria that cause

separately (12,4).

leakage of metabolites that inactivate the

The interaction showed the formation of

bacterial

enzyme

systems.

Damaged

a bridge at or near the intersection of two

cytoplasmic membranes can prevent the

inhibitory zones or no longitudinal inhibitory

entry of nutrients needed by the nutrients the

zones formed. The combination is said to be

bacteria produce energy. It causes bacteria to

synergistic if it created like a bridge at or near

experience growth retardation and even cause

the intersection point of two inhibitory zones,

bacterial death (6).

or an obstacle to growth which is the

Aloe vera contains active substances

combined effect of the two antimicrobial

such

as

anthraquinone,

tannins

and

agents (13).

flavonoids.

Anthraquinone

binds

with

Based on observations of the results of a

nucleic acids and forms a complex that can

combination of inhibitory test of Chinese

damage the acid (DNA and RNA) of

Petai leaf extract and aloe vera on E. coli

inhibited bacteria. Tannin as antibacterial

(mm) bacteria, the concentration was 25

works by inactivating adhesin so that bacteria

mg/mL producing a zone of inhibition 6.3 ±

cannot stick to host epithelial cells. Aloe vera

0.58 mm, a concentration of 50 mg/mL

also contains flavonoids, which will cause

resulted in an inhibition zone of 8, 3 ± 0.58

lysis and inhibit the formation of cell walls.

mm, concentration of 75 mg/mL produces

This mechanism causes aloe vera to kill or

inhibition

mm,

inhibit the formation of bacteria (15,16,17).

concentration of 100 mg/mL with inhibition

Flavonoids are efficient to inhibit the

zone

10.3

±

0.58

zone 11.7 ± 0.58 mm. Positive control using

growth

the antibiotic Chloramphenicol obtained the

Flavonoids are polar compounds which are

biggest inhibition zone compared to other

easy to penetrate the peptidoglycan layer.

concentrations of 27.2 ± 1.26 mm.

Furthermore, the Gram-positive cell wall

According to Davis stout in A'lana (14),

contains

of

Gram-positive

polysaccharides

bacteria.

(acidic

the strength criteria for the combination of

polysaccharides) is a water-soluble polymer.

Chinese

vera

This cell wall functions as the media of

concentrations of 25 mg/mL and 50 gr/mL

positive ions transport to get in and out. This

categorized as moderate, at a concentration of

solubility indicates that the Gram-positive

75 mg/mL and 100 mg/mL classified as

cell wall is polar. Flavonoids disrupt cell wall

strong criteria.

function. This results in cell lysis (3,20,21).

petai

leaves

and

aloe

The mechanism of bacterial inhibition of

growth of bacteria E. coli by damaging the

tannin compounds is by protein reproduction,

Ina J Med Lab Sci Tech 2020; 2(2): 60-67

65

Petai China has saponins to inhibit the

Putra Rahmadea Utami, et al.

ie through reaction with cell membranes,

vera inhibited the growth of E. coli. The most

enzyme inactivation and inactivation of

efficient extract concentration was 100

genetic material functions, in addition to

mg/mL. The combination of extracts can

inhibiting the reverse transcriptase enzyme

provide effective results in inhibiting the

and DNA topoisomerase so that bacterial

growth of bacteria, especially E. coli bacteria.

cells cannot be formed. Saponins which can
provide antibacterial activity are avenacin.
Saponin

compounds

cytoplasmic

will

membrane

damage

and

kill

CONFLICT OF INTEREST

the

There are no conflicts of interest.

cells

(20,22,23).

ACKNOWLEDGMENT(S)
This study is supported by Ministry of

CONCLUSIONS
The conclusion of this research is there is
an interaction in the combination of ethanol

Research, Technology and Higher Education
of Indonesia through the research funding for
beginner lecturers in 2019.

extracts of Chinese petai leaves with aloe

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