Division of San Jose City

San Jose City National High School

Calaocan, San Jose City, Nueva Ecija

BIOASSAY OF AIR DRIED AND FRESH LEAVES FROM SALUYOT (Corchorus

olitorius) IN SOAP

An Investigatory Project
Presented to the Board Judges
As Entry for the 2018 School Science & Technology Fair
Life Science- Group Category

Project Proponents:

Martin Lawrence L Asinas

John Edison V. Brillo
Acel Deane D. Ciron

Research Adviser:
Jonnalyn G. Pablo

Consultant:
Hanson T. Villanueva

September 2018

1
 INTRODUCTION

Background of the Study

In the present generation, people are very conscious of their physical appearance. This is

why many companies produce different beauty products, such as soap, lotion, facial cleanser,

and toner, that promise to help make the skin lovely and glowing. But many pollutants in the

environment make the skin unhealthy. One of the indications of an unhealthy skin is having

many bacteria. Soap is an essential cleansing agent in our daily life. Soap (sodium salt of fatty

acid) is the oldest known surfactant and has been used since the Egyptian era (Kaoru, Ainie et al.

1998).

According to Jay Nelz, 2016, Saluyot leaves are rich in vitamins, carotinoids, calcium,

potassium and dietary fibers. It is acclaimed also for its anti-aging benefits. Studies have shown

that this green, leafy vegetable is also rich in the following nutrients beta-carotene which is best

for promoting good eyesight, iron for healthy red blood cells, calcium for strong bones and teeth,

and vitamin C for smooth, clear skin, strong immune cells, and fast wound-healing. Coumarin is

a chemical compound present in many plants especially in saluyot. This substance contains

antifungal and antibacterial property according to Cowan MM, 1999. The Vitamins A, C and E

present in saluyot “sponge up” free radicals, scooping them up before they can commit cellular

sabotage (AbouZeid AHS. 2002). Saluyot contains vitamin E and other antioxidants. It is said to

prevent wrinkles and promote youthful looking skin. (Azuma K et. Al, 2016).

Nowadays, people are finding ways to cleanse skin in a manner that the usage of

chemicals are reduced. This marked the start of producing a soap which is eco-friendly and

natural specifically, soap with saluyot leaves.

2
Statement of the Problem
This study aims to determine the antibacterial property of Saluyot (Corchorus olitorius)

leaves in soap.

Specifically:

1. It aims to produce a soap containing saluyot that will inhibit the growth of bacteria S.

aureus, E. coli and C. albicans.

Hypothesis

Ho: There is no significant difference between treatments containing saluyot in soap in terms

of its zone inhibition of bacteria.

Significance of the Study

Saluyot can grow almost anywhere in the Philippines. It is a hardy plant that is resistant

to pests and requires a little care. It can be found in the wild as well in farms. The abundance of

saluyot is the reason why the researchers chose it. It is cheap and easy to acquire.

This study not only will help lessen bacteria around your body but will also decrease

waste in the surroundings. Knowing its benefits, the researcher came up with an idea to make an

alternative soap which is cheaper and not harmful to the environment.

Making soap out of Saluyot (Corchorus olitorius) aims to determine the capability and

effectiveness of this plant as a component in antibacterial body soap. In this project, the

researchers are determining how it kills the bacteria which is present on the human’s skin. On the

other hand, the leaves of the saluyot will be the one responsible for cleansing the skin.

3
Scope and Limitation of the Study

This research was conducted to determine the antibacterial property of Saluyot

(Cochorus olitorius) which was mixed in soap. The researchers tested gram - positive

microorganisms (e.g. Staphylococus aureus, Candida albicans) and a gram - negative bacteria

(e.g. Escherichia coli). This study only covered the use of air-dried and fresh saluyot leaves

against these pathogens and only aims to determine which treatment was effective. The

proponents excluded the use of other chemicals and the product is only intended for the use as

antibacterial bath soap. The researchers conducted this study from January- September 2018 at

the laboratory room of San Jose City National High School.

METHODOLOGY

Preparation of Saluyot leaves and Oils

The researchers obtained the saluyot leaves at zone 6, barangay A. Pascual, San Jose City

Nueva Ecija. The procedure started with the gathering of 100g saluyot leaves. Saluyot leaves

were dried in room temperature (20-25ᵒC) for 25 days. Fresh leaves were rinsed using distilled

water which is appropriate to use because it has no contaminants and chemicals. Both fresh and

air-dried leaves were crushed using a mortar and pestle and was weighed using a weighing scale.

Olive oil, palm oil, coconut oil, and eucalyptus oil were measured using a graduated cylinder.

4
After measuring, olive oil (547.11ml), palm oil (473.18ml), coconut oil (207.02ml), and

eucalyptus oil (29.57ml) were mixed in a bowl for saponification.

Saponification

First, the lye solution was prepared by mixing the weighed dry lye with the amount of

water. Hard water was not used for cold process soap making because the unwanted substance

within the water could take part in the reaction (Burts S. 2004). The dry lye was measured in a

small container with a lid. After mixing the components the lye was poured into the water not the

water into the lye. The lye and water 3:1 (473.18ml:171.53ml) was mixed till the lye was

dissolved.

The lye was stirred constantly until the temperature dropped at 35ᵒC. After the desired

temperatures met, the lye was slowly poured into the oils and fats mixture while stirred

vigorously. The lye solution was transferred into a bowl to make it easier to slowly pour the

chemical into the oils. The solution was stirred and rather briskly in order for all the necessary

molecules to interact. After trace is reached, other ingredients were added including essential oils

or fragrances and other various nutrient. The solution was then transferred to a molder for the

molding of soaps and the mixing of crushed air-dry and fresh leaves. Soap were stored in room

temperature for 4 weeks for the solidification.

Testing the antibacterial effect

Determining antimicrobial properties of the saluyot leaves in soap was performed in the

laboratory of Saint Mary’s University in Bayombong, Nueva Vizcaya using the agar disc

diffusion technique using antibiotic assay discs. The consultant, Mr. Hanson T. Villanueva

5
conducted a test against two Gram-positive microorganisms (e.g. Staphylococcus aureus and

Candida albicans) and a Gram negative bacteria (e.g. Escherichia coli). Plates was incubated at

37°C for 24 to 48 hours. Disc diffusion method was used to screen the antibacterial activity. In

vitro, antibacterial activity was screened by using nutrient Agar (NA). The NA Plates was

prepared by pouring 15-20ml of Nutrient Agar media into sterile petriplates. The plates were

allowed solidifying for 5 minutes. The inoculum suspensions were swabbed uniformly and

allowed to dry for 5-10 minutes. The extracts were loaded on 6 mm sterile disc. The load disc

was placed into the surface of medium and the compound is allowed to diffuse for 5 minutes and

the plates will keep for incubation at 37ᵒC for 24 hours. At the end of incubation, inhibition

zones formed around the disc and was measured using digtal vernier caliper. The test was

performed in triplicate.

Flow Chart

Preparation of Saluyot leaves

Saponification

RESULTS AND DISCUSSION

Testing the antibacterial effect
6
 Figure 1. Flow
RESULTS ANDChart of the study
DISCUSSION

Table 1. Zone of Inhibition (obtained in mm) Soap A

Test Zone of Mean

Organism Inhibition
S. aureus 6mm 6mm 6mm 6mm

E. coli 12.3mm 9.8mm 10.7mm 10.9mm

C. albicans 6mm 6mm 6mm 6mm

Table 1 shows the result for the zone of inhibition of S. Aureus, E. Col and C. Albicans

using the air-dried saluyot leaves in soap. The mean zone inhibition for S. Aureus has a value of

6mm, E. Coli with 10.9mm, and C. Albicans with 6mm respectively. Statistically the means

values are not significantly different from each other, thus T1 containing air-dried saluyot leaves

is the most active in inhibiting the growth E. Coli active and is partially active on S. Aureus. It

does not inhibit the growth of C. Albicans.

Table 2. Zone of Inhibition (obtained in mm) Soap B

Test Zone of Mean

Organism Inhibition
S. aureus 13.4mm 10mm 10.7mm 11.4mm

E. coli 15.6mm 14.4mm 12.3mm 14.1mm

C. albicans 6mm 6mm 6mm 6mm

7
 Table 2 shows the results for the zone of inhibition of S. Aureus, E. Coli and C. Albicans

using fresh leaves in soap. The mean zone of inhibition for S. Aureus has a value of 11.4mm, E.

Coli with 14.1mm and C. Albicans with 6mm respectively. Statistically the mean values are not

significantly different from each other, thus T1 containing fresh leaves is partially active in the

inhibition of E. Coli. It is inactive in both S. Aureus and C. Albicans.

Test Zone of Mean

Organism Inhibition
S. aureus 6mm 6mm 6mm 6mm

E. coli 12.3mm 9.8mm 10.7mm 10.9mm

C. albicans 6mm 6mm 6mm 6mm

Table 3. Zone of Inhibition (obtained in mm) Soap C

Table 3 shows the results for the inhibition of S. Aureus, E. Coli, and c. Albicans

using the soap with no saluyot leaves. The mean zone of inhibition for S. Aureus has a value of

6mm, E. Coli with 10.9mm, and C. Albicans wit 6mm respectively. Statistically the means

values are not significantly different from each other, thus T3 with no leaves is partially active in

the inhibition of E.Coli. It is inactive in both S. Aureus and C. Albicans.

8
 Treatments/test S. aureus E. coli C. albicans Mean

organism
Treatment 1

(Soap A-air 11.4mm 14.1mm 6mm 10.5mma

dried)
Treatment 2

(Soap B-fresh) 6mm 10.9mm 6mm 7.63mmb

Treatment 3

(Soap C- 6mm 10mm 6mm 7.33mmc

control)
Table 4. Mean per Treatment of Soap’s Zone of Inhibition (obtained in mm) Soap

Statistically, table 4 shows that treatment 1 (air-dry) obtained the highest mean,

Treatment 2 (Fresh leaves) has the second highest mean, and Treatment 3 (Control) has the

lowest mean among the treatments. Treatment 1 containing air-dried Saluyot leaves got the

highest mean value, this can be attributed to the fact that coumarin compounds found in the

leaves were much more concentrated due to the isolation of other compounds which were

obtained from air-drying process (J Mǖller, A Heindl – Frontis,2006).

SUMMARY, CONCLUSIONS, AND RECOMMENDATIONS

9
 This chapter describes the purpose of the study, summarizes the results and findings of

the study, and offers some recommendations for future research regarding said topic.

Summary

The three treatments which included soap with saluyot air-dried leaves, fresh leaves, and

absence of leaves were subjected to disc diffusion method as eradicant against the Escherichia

coli, Staphylococcus aureus, and Candida albicans bacteria. As for Staphylococcus aureus, the

highest measure of zone of inhibition obtained was the Treatment 1 (Air-dried). However,

Treatment 2 (Fresh leaves) didn’t inhibit the growth of the bacteria, followed by Treatment 3

(Control), which also didn’t show a zone of inhibition. Similar to the Staphylococcus aureus

case, Treatment 1 (Air-dried) exhibited the widest diameter of the zone of inhibition against

Escherichia coli. However, Treatment 2 (Fresh leaves) and Treatment 3 (Control) partially

showed a zone of inhibition against the same bacteria. As for Candida albicans, none of the three

treatments exhibited a zone of inhibition. Treatment 1 (Air-dried) obtained the highest mean,

Treatment 2 (Fresh leaves) obtained the second highest and Treatment 3 (Control) has the lowest

mean among the three treatments. T1 containing air-dried leaves possessed the greatest

antibacterial property against E. coli and S. aureus even greater than the soap containing fresh

leaves and the control. The remaining treatments showed zone of inhibition against pathogens

but not as effective as the treatment containing air-dried leaves. T1 possessed greater

antibacterial property against Escherichia coli among other bacteria. The classification of T1

(Air-dried) using the disc diffusion method was susceptible against S. aureus and was

immediately susceptible against E. coli. T2 (Fresh leaves) and T3 (Control) was partially

susceptible against E. coli. Both T2 and T3 was resistant to S. aureus. All treatments were

resistant to C. albicans.

10
 Conclusion

Based on the results of the study, it can be concluded that only T1 (Air-dried) has a great

effect in inhibiting the growth of pathogens Escherichia coli, Staphylococcus aureus and is

resistant to Candida albicans. While the soap containing fresh leaves and the control soap have

antibacterial efficacy towards E. coli but are resistant to S. aureus and C. albicans.

Recommendations

1. Use different parts of saluyot for comparison of strength in anti-bacterial inhibition

towards pathogens.

2. The different treatments should undergo testing conducted by Bureau of Food and Drug

Administration (BFAD) so that the safety of the soaps towards skin irritations and other

side effects will be approved.

3. Use commercially available soap for comparison between treatments in terms of strength

in anti-bacterial inhibition towards pathogens.

4. Determine the antibacterial efficacy of Saluyot in different ointment including liquid

soap, lotion and cream.

11
 BIBLIOGRAPHY

Burts S. 2004. Essential oils: their antibacterial properties and potential applications in foods- a

review. International Journal of Food Microbiology. 94: 223-253.s

Cowan MM. 1999. Plant Products as Antimicrobial Agents. Clinical Microbiology Reviews. 12:

564-582

Tulio Jr AZ, Ose K, Chachin K, Ueda Y. 2002. Effects of storage temperatures on the

postharvest quality of jute leaves (Corchorus olitorius L.). Postharvest Biology and Technology.

26:329-338

AbouZeid AHS. 2002. Stress metabolities from Corchorus olitorius leaves in response to

certain stress agents. Food Chemistry. 76: 187-195

Kaoru, T. (1998). Surface activity: Principles, phenomena and application, p. 21-22. Academic

Press, San Diego.

J Mǖller, A Heindl – Frontis,2006- library. wur .nl. Drying of medicinal plants.

Jay Nelz, 2016. Top Herbal Plant: Benefits of Saluyot (Corchorus olitorius)

Azuma K, Nakayama K, Koshioka M, Ippoushi K, Yamaguchi Y, Kohata K, Yamauchi Y, Ito H,

Higashi H. Phenolic antioxidants from the leaves of Corchorus olitorius L. J Agric Food Chem

1999; 47:3963-3966.

12
APPENDICES

Appendix
Appendix Figure Figure 2. of
1. Preparation Weighing of lye
Materials

Appendix Figure 4. Weighing of water

Appendix Figure 3. Weighing of oil

13
Appendix Figure 5. Rinsing
Appendixof Figure
Saluyot6. Pounding of Air-Dried
leaves

Appendix
Appendix Figure 7. Pounding Figure 8. Mixing the lye
of Fresh
Leaves

14
Appendix Figure 9. Mixing the oils Appendix Figure 10. Mixing the water

Appendix Figure
Appendix Figure 12.
11. Putting
Mixing solution
the solution
in a
molder

15
Appendix Figure 13. Adding the air-dried
and fresh saluyot leaves to the lye solution

Appendix Figure 14. Actual soap product

16
I approve this research plan.
Teacher’s Name: JONNALYN G. PABLO

Teacher’s Signature:

17