Ashitaba as an Alternative to Commercial Hand Sanitizer
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 their respective media by placing 0.2ml of each. The inoculated agar was then poured to individual sterile plates. D. Extraction from Leaves
The extraction was done through the use of a blender. Five leaves of the same size were placed in it. They were then crushed until the yellowish sap was produced. Preparation of a filter paper or cloth was the next step. Pour the extract into the paper or cloth. Squeeze tightly. The filtered extract was put in reagent bottles and kept refrigerated until use. E. Determination of the Activity of the Extract on the Chosen Bacteria
The antibacterial property of leaves extracts obtained from Angelica keskei koidzumi was determined using Kirby-Bauer disk method. Using this method, a culture medium, nutrient agar, was uniformly and aseptically inoculated with the test organism and then filter paper discs soaked in 10 ug/mL of ashitaba extract are placed on the medium. Incubation was done for 2 days at 37 0C. The zones of inhibition were then measured. F. Formulation of Hand Sanitizer
Fifty milliliters of water was poured in the jar. One milliliter of xanthan gum powder was prepared. This was then sprinkled over the water little by little and at the same time, the mixture was whisked
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 their respective media by placing 0.2ml of each. The inoculated agar was then poured to individual sterile plates. D. Extraction from Leaves
The extraction was done through the use of a blender. Five leaves of the same size were placed in it. They were then crushed until the yellowish sap was produced. Preparation of a filter paper or cloth was the next step. Pour the extract into the paper or cloth. Squeeze tightly. The filtered extract was put in reagent bottles and kept refrigerated until use. E. Determination of the Activity of the Extract on the Chosen Bacteria
The antibacterial property of leaves extracts obtained from Angelica keskei koidzumi was determined using Kirby-Bauer disk method. Using this method, a culture medium, nutrient agar, was uniformly and aseptically inoculated with the test organism and then filter paper discs soaked in 10 ug/mL of ashitaba extract are placed on the medium. Incubation was done for 2 days at 37 0C. The zones of inhibition were then measured. F. Formulation of Hand Sanitizer
Fifty milliliters of water was poured in the jar. One milliliter of xanthan gum powder was prepared. This was then sprinkled over the water little by little and at the same time, the mixture was whisked