Synthesis of Handmade Lemon Soap and Its

Antibacterial Activity Test Against Staphylococcus

aureus and Escherichia coli

Winarto Haryadi
Dept. of Chemistry, Universitas gadjah Mada
Yogyakarta
Lemon (Citrus limon L)
• Lemon is a herbal plant of rutaceae family that contains alkaloids
• The main alkaloids compound is saponin which is active as
antibacterial, anticancer and antifungal (Mohanapriya et al, 2013).
• Lemon juice can inhibit the growth of pathogens that cause diarrhea,
in this case, the Enterotoxin Escherichia coli (ETEC) bacteria (Ekawati,
2019)
 Lemon soap
• Lemon soap is made from saponification of vegetable oils and strong
bases with the addition of lemon juice.
• Saponification reaction :
 Lemon soap as an antibacterial
• Alkaloids in the lemon soap are predicted active as antibacterial.
• The in vitro antibacterial activity can be investigated by Kirby-Bauer
Diffusion methods against Staphylococcus aureus ATCC 25923 (Gram
positive) and Escherichia coli ATCC 11229 (Gram negative)
The making of lemon soap

The mix. of Vegetable oils;

121 g olive, 106,5 g palm, Added 1% of lemon juice,
106,5 coconut and 15 g Stirred until the soap was
stirred until completely
castor oil perfectly formed at room
mixed, poured into the
temperature or
+ mold curing process for 4
saponificanion completed
50 g of NaOH in 135 mL of weeks
water
Antibacterial test
Bacterial isolates (E coli ATCC 11229 and S.
aureus ATCC 25923 were grown on
Nutrient Broth (NB) medium during the 24
h incubation period at 37°C
The bacterial suspension was inoculated on
Mueller Hinton Agar (MHA) plate medium
according to the spread plate method. Blank paper
discs were dipped for 5 min. on various treatments,
and then placed on the surface of MHA plate.

The treatments were performed by the additions of 5% lemon

soap with the concentration of 2.5%, 5%, 7.5%, and 10% for
each. The positive control was chloramphenicol (10 ppm) and the
negative control was aquadest only. The plates were incubated
for 24 h at 37°C and the diameter of the clear zone was
measured using the sliding term at 6, 12, 24 hours.
Handmade lemon soap
Antibacterial activity screening using diffusion
method (Kirby and Bauer Method)
Antibacterial test against E. Coli
Inhibition Diameters

30

25

20
diameter (mm)

15

10

0
0% (- control) 2,5% 5,0% 7,5% 10,0% Klor 10% (+ control)
6 hours 0 28.76 25.11 22.79 16.50 14.27
12 hours 0 7.46 24.96 23.44 15.16 13.38
24 jam 0 7.71 16.54 10.07 12.81 12.30
Concentration (%)

6 hours 12 hours 24 jam

Antibacterial test against S. Aureus
inhibition diameter of antibacterial test of lemon soap against S.aureus

30.00

25.00

20.00
diameter (mm)

15.00

10.00

5.00

0.00
0% 2,5% 5,0% 7,5% 10,0 Klor 10
6 jam 0.00 20.51 24.57 22.59 19.94 26.64
12 jam 0.00 15.21 19.10 18.84 16.28 14.43
24 jam 0.00 13.59 18.86 18.80 15.38 10.86
concentration

6 jam 12 jam 24 jam

conclusion
• Lemon soap is more active as an antibacterial than chloramphenicol
both for Gram positive (S. aureus) and Gram negative (E. coli)
bacteria. At 6 hours incubation, the lemon soap with a concentration
of 2.5% had the highest activity against E.Coli but chloramphenicol
still had a higher activity against S. aureus. At 12 and 24 hours
incubation, 5% soap concentration had the highest activity against
E.coli and S. aureus.