Antibacterial Activity of Santol (Sandoricum koetjape) Leaf Extract Against

Staphylococcus Aureus

Research Plan

A. Question or Problems being addressed

A certain research from the Bulletin Pharmaceutical Research showed that E.coli E.

aerogenes, Streptococci and Staphylococcus aureus in the percentage of 23.53%, 23.53%,

17.65%, and 35.30% respectively were found on phones. (Verma, D.K., 2015)

But aside from smartphones, Staphylococcus spp can also be found in the human skin

especially in the hands and on our faces which causes pimples build up, Unakal (2019).

According to Otto (2010), staphylococci are the most abundant skin-colonizing bacteria and the

most important causes of nosocomial infections and community-associated skin infections.

The ingestion of these bacteria can be very detrimental to one’s health. Disinfecting

these microorganisms is needed to avoid its growth that may cause harmful diseases.

B. Goals / Expected Outcome / Hypothesis

The main goal of this research is to discover the efficacy of the Santol leaf extract

against Staphylococcus aureus with the significant components found in Santol leaf extract that

are utilizable for the elimination of bacteria. The study aims to determine the effectivity of Santol

leaf extract in dissolving these bacteria, specifically the Staphylococcus aureus, found in skins as

well as contaminated cellular phones

C. Procedures
Collection of Santol Leaves

Santol leaves will be collected from a researcher's residence; the samples will be washed

with water thoroughly and will be air dried for 3 days. It will be powderized using a blender.

Preperation and Extraction of Santol (Crude) Leaf Extract

Powderized santol will be macerated with ethanol for 24-48 hours. The solvent will be

removed through a Rotatary Evaporator.

Inoculum Preparation

An inoculating loop will be used to scrape off enough bacteria sample from its colony

and will be mixed with a 20 mL distilled water. The mixture is called an inoculum. Petri dishes

will be prepared.

Disc Diffusion Assay

The nutrient agar will be added to the petri dishes using the pipette followed by the

inoculum. After that a spreader will be sterilized under the process of flaming, then it will be

cooled and will be used to spread out the bacteria in the dish evenly. The improvised antibiotic

sensitivity discs will be then soaked in the santol leaf extract and will be placed with the petri

dishes. 3 discs will be placed in every petri dish with 3 cm apart. After closing the petri dishes,

the lid of the covers will be sterilized to kill excess microorganisms.

Storing of the Treatments

 After the Disk Diffusion Assay procedure, the Treatments will be then stored in the oven

dryer for about 12 hours so that the bacteria can inhibit the petri dishes.

Gathering of Data According to the Zone of Inhibition

After 12 hours, the petri dishes will be then pulled out and it is ready for measuring.

Using the Vernier Caliper, the zone of inhibition of each treatments will be measured in

millimeters and the data were recorded.

Gram Staining

A smear will be prepared from the given bacterial culture and were placed in two glass

slides. Two slides will be placed on the wire staining screen and will be flooded with methylene

blue for 1 minute. After staining for 1 minute, the methylene blue will be washed with tap water

and the excess water was drained off. After that, the two glass slides with smear will be then

flooded with iodine for another minute. After one minute, these will be washed with tap water

and will be lightly blotted with tissue paper to remove excess water but it will be not completely

dried. The slides will be then tilted and were decolorized with 95%alcohol for about fifteen

seconds. The slides will be washed again with tap water. The smears were then counter-stained

with carbol fuchsin for about 30 seconds and will be washed with tap water. The slides will be

examined under the microscopes and will be identified if it will be gram positive or gram

negative. The structure and color or the bacteria Staphylococcus will be observed.

Proper Disposal

For standard sanitary measures, the researchers will see to it that the used treatments and

used experimental materials will be properly disposed.

D. References

A., A. El-Oqlah, and A. M. Mahasneh. 1992. Herbal Plants in the Traditional Medicine of

Bahrain. Retrieved from //www.plantsforuse.com/species/poaceae-eleusine

Barasa, et al., (2015). Isolation and Characterization of Bacteria from Mobile Phones of Students

and Employees at University of Gondar, Ethiopia. Bulletin of Pharmaceutical Research.

5(3):96-100

Blanco, M. (2016). Santol Sandoricum koetjape (Burm.f.) Merr. LOLLY FRUIT . Retrieved

January 14, 2018 from http://www.stuartxchange.org/Santol.html

Bootle, K.R. (1983). Wood in Australia. McGraw-HILL: Sydney. Retrieved from

https://www.homeaffairs.gov.au/trav/ente/brin/can-i-bring-it-back/Can-I-Bring-It-Back-

Market-Goods-And-Shopping/Can-I-Bring-Wooden-and-Woven-Items-Back

Boshell, P. (2013). Your Mobile Phone is Dirtier than You Think. Retrieved January 13, 2018

http://info.debgroup.com/blog/bid/290652/your-mobile-phone-is-dirtier-than-you-think