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Ashitaba as an Alternative to Commercial Hand Sanitizer

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CHAPTER III
METHODOLOGY

This chapter includes all the materials and procedures that were properly followed to gather the needed information and necessary data for the study.

A. Sources of Test Organisms
The bacterial isolates (Staphylococcus aureus, Escherichia coli and Streptococcus) used in the experiment were obtained from Institute of Biological Science, University of the Philippines Los Baños, Laguna. B. Maintenance of Microbial Cultures
Bacterial isolated were subculture on slants of Nutrient Agar (NA). Incubation of the subcultured specimens for bacteria was done for 7-14 days in Isolation Room. C. Media Preparation for Assay
Three tubes with nutrient agar were prepared. The cultures were inoculated onto …show more content…

G. Testing the Effectiveness of the Formulated Hand Sanitizer
Using two sites of the arm, the effectiveness of the hand sanitizer was tested. Swab head was moisten with sterile water and rubbed three times on the surface in rotating motion. This swab head was put in the tube. The tube was then shaken making 50 complete cycles of 15 cm in 10 seconds. A 1:10 dilution of the rinse solution was prepared. On Tripticase soy agar, the plate 1.0 and 0.1mL portions of the resulting suspension was poured. The plates were incubated at for 370C 1-2 days. The number of colonies per mL of the rinsing solution was calculated. Using a specific formula, the microbial load of the 4cm2 surface of the skin was determined.
The second area which was marked was wet and rubbed with sanitizer for 15 seconds. This area was lathered for 60 seconds without soap and rinsed for 15 seconds with sterile water. Some steps are repeated to determine the CFU/cm2 of the second washed area. H. Data Analyses
Extracts from leaves of Ashitaba were applied on bacterial cultures. Detection of antibacterial activity was done after 7 days. The plates were compares in terms of the diameter of zone of inhibition. Hand sanitizer was then formulated using the extract. This was tested on a specific part of the skin. The abundance and type of microbial load of treated and untreated surface of the skin were compared.

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