ADVINCULA, Trisha Joy D. REBELLO, Kate DE LEON ,Christine DELA CRUZ, Patricia 9- St. Andrew Mr.

John Mark L. Doria

INVESTIGATORY PROJECT Papaya (Carica papaya) Antibacterial Agent ABSTRACT: This study determined the efficacy of Carica papaya as an antibacterial agent. The two sets of experiment were tested using contaminated slides (marked A & B). The products on experiments on A & B are used to test if it could kill germs on contaminated slides (marked A & B). It was noted that almost 85% of the germs on slide A has been killed with product A while almost 65 % of the germs on slide B has been killed with product B. The conclusion of the results is that the mixture of papaya extract and water has antibacterial properties. Adding aloe gel and alcohol made the product better and convenient in usage. Further test was done and showed that commercialized hand sanitizer killed almost 80% of germs on the glass slide thereby proving that adding papaya extracts added in the antibacterial function of the hand sanitizer ingredients. Chapter I I. INTRODUCTION: Today, one of the major issues in our society are sicknesses and diseases like influenza, conjunctivitis, sore throat and other diseases that affects our daily routine and oftentimes inconvenient to us. We would always find the best way to protect ourselves. Department of Health and even TV commercials would advertise or remind us that frequent washing of hands with soap and water would prevent us from having such diseases, but water and soap is not always available especially if we are outside our houses. Alcohol is proven and tested to kill germs, bacteria and other harmful micro-organisms but can dry hands, and commercial sanitizers are readily available in the market, but how sure are we that this will not cause harm to us, so we decided to

make an eco-friendly and effective hand sanitizer that can be an alternative to the commercial hand sanitizer. II. STATEMENT OF THE PROBLEM: 1. Will the papaya fruit extracts be a good antibacterial agent when incorporated with hand sanitizer ingredients? 2. Will the solution do the antibacterial functions? 3. Is the product competent to the commercial hand sanitizers? III. FORMULATION OF THE HYPOTHESIS: 1. The finished product might spoil due to incorporation of the papaya fruit extracts. 2. The product might be too aqueous because of the amount of alcohol. 3. The product might be too sticky due to the aloe Vera gel. IV. SIGNIFICANCE OF THE STUDY: This study promotes awareness on hygiene and sanitation. It will also give confidence to the people who have undergone this study by letting them know how to make topical antibacterial solutions in absence of commercial products. It could give them knowledge that they could share to other people to prevent spread of diseases. V. SCOPE AND LIMITATIONS This study of the Carica papaya fruit extract focuses mainly on the determining its antibacterial properties. Such study includes researches, gathering of materials and performing experiments. Evaluation and conclusion of results is also needed to come up with conclusions and recommendations. The limitations of this study are on deriving exact values such as the exact formulation of the solution, determining the types of bacteria it could kill, quantity of bacteria that could be killed per volume of the solution and how long the product protect the user once it is applied. VI.DEFINITION OF TERMS: INFLUENZA- a highly contagious virus infection that affects the respiratory system.

CONJUNCTIVITIS- inflammation of the conjunctiva, which becomes red and swollen and Produces a watery or pus-containing discharge. VISCOSITY- the property to flow in a fluid or semi-fluid. EXTRACT-a preparation containing the pharmacologically active element of a substance. Chapter II Review of Related Literatures We know the hand sanitizers are good, but are we right? Is it good or not? What we know is good for our health. Do you know we must look at the label of it as told by a professor to read my article? The inscription "Check the bottle for active ingredients. It might say ethyl alcohol, ethanol, isopropanol, or some other variation, and those are all fine. According to a professor only 40% of element of hand sanitizer was the gels ingredients. The hand sanitizer with 60% Alcohol can not kill the germs. The alcohol or hand sanitizers will not kill the germs such a chance so we are still getting sick. Others use high quality alcohol based gels instead of soap and water on their Hands. For we are able to avoid carrying disease germs better we wash hands with soap. The hand sanitizer is as ever from soap and water? We all know that anyway better hand washing. The alcohol based gels are proven resistance to germs temporary substitute when soap and water are not available. In a study at Boston families with children who use or use hand sanitizer within 5 months, gastrointestinal stop the spread of infections by 59%. In another study families use hand sanitizer about 5 times a day will stop the risk for colds by 20% in as the families who scrubbed less often. For sure we can make sure we have the hand sanitizer that we have used and least 60% alcohol for it to be effective. Hand sanitizers can kill 99.9% germs. According to the study the hand sanitizers are deadly for germs. As of hand sanitizers, soap can be effective. According to other fatality rates do not necessarily need the manufacturers to kill 99.9% of germs. Regulations do not require them to disclose. Which they exterminate bugs, just that the products are effective against a representative sample of microbes. Yet by killing other more common bugs. They can claim 99.9% effectiveness. The FDA warns about not buying hand sanitizers "that claim to prevent infection from MRSA, E. coli, salmonella, flu, or other bacteria or viruses." Although the FDA like that saying not to use or buy said they still can still use hand sanitizer when there is no water available. "FDA has not approved any products claiming to Prevent infection from MRSA, E. coli, Salmonella, or H1N1 flu, Which a consumer can just walk into a store and buy," Do not

count on hand sanitizers for 100% protection from anything . Do wash your hands often. And when you cannot wash your hands, use hand sanitizers do. Even the FDA agrees they get rid of a lot of the germs that are on your hands The CDC advice specifically says alcohol-based hand sanitizer help protected against MRSA and other germs. During flu season, the CDC warns Americans to continually Prevent Flu by using hand sanitizers when soap and water are not around.

1. Antibacterial substance from Carica papaya fruit extract. J Nat Prod. 1982 Mar-Apr;45(2):123-7. Emeruwa AC.

Ripe and unripe Carica papaya fruits (epicarp, endocarp, seeds and leaves) were extracted separately and purified. All the extracts except that of leaves produced very significant antibacterial activity on Staphylococcus aureus, Bacillus cereus, Escherichia coli, Pseudomonas aeruginosa and Shigella flexneri. The MIC of the substance was small (0.2-0.3 mg/ml) for gram-positive bacteria and large (1.5-4 mg/ml) for gram-negative bacteria. The substance was bactericidal and showed properties of a protein. Other proteins previously found in C. papaya did not show antibacterial activity.

2. Chemical Profile of Unripe Pulp of Carica papaya O.I. Oloyede Department of Biochemistry, University of Ado-Ekiti, Ekiti State, Nigeria.

Unripe pulp of Carica papaya was screened to test for the presence of certain phytochemicals. Chemical composition of the pulp were determined. Phytochemical screening of mature unripe pulp of Carica papaya (dry weight) showed the presence of saponins and cardenolides while chemical analysis revealed the presence of potassium (223.0mg/100g) as well as sodium, calcium, iron, phosphorus, zinc, copper, magnesium and manganese in considerable quantities. Proximate analysis of the pulp showed that it contained starch (43.28%), sugars (15.15%), crude protein (13.63%), crude fat (1.29%), moisture (10.65%) and fibre (1.88%). All these results indicate that the pulp of mature unripe Carica papaya contains nutrients and mineral elements that may be useful in nutrition. The presence of some phytochemicals like saponins and cardenolides explained the astringent action of the plant encountered in the numerous therapeutic uses.

Materials and Methods Fresh, unripe mature fruits of Carica papaya (Variety T. solo) were obtained from National Horticultural Research Institute (NIHORT), Ibadan, Nigeria. The fruits were peeled, seeds removed and the pulp cut into small pieces, sundried and finely powered with an electric grinder. The powered material was stored in properly sealed bottles at 10 deg.Cel. in the refrigerator. Proximate analysis, mineral composition and phytochemical analysis were carried out on dried samples of unripe pulp of Carica papaya. Proximate analysis: Proximate analysis were carried out according to the procedure of Association of Official

Analytical Chemist (A.O.A.C .,1990). This constitute the class of food present in samples such as carbohydrate, protein, fat, free sugar, starch fibre, ash content and moisture content. Mineral composition: This shows the quality of organic and inorganic acid metals present in the samples. The minerals determined are Sodium, Calcium Iron, Phosphorus, Potassium, Zinc, Copper, Magnesium, Manganese, Cobalt, Cadmium, and Lead. The samples were dry ashed at 550 deg.C. The ash was boiled with 10ml of 20% hydrochloric acid in a beaker and then filtered into a 100ml standard flask. It was made up to the mark with deionized water. The minerals were determined from the resulting solution using Atomic Absorption spectroscopy (Pye unican sp9, Cambridge, UK). Phytochemical analysis: Phytochemical screening procedures carried out were adapted from the previous work on plant analysis (Odebiyi and Sofowora, 1979; Sofowora and Odebiyi, 1978. This analysis determines the biologically active nonnutritive compounds that contribute to the flavor, colour, and other characteristics of plant parts. Examples of these are alkaloids, tannins, 1 . cardiac glycosides (e.g. cardenolides), anthraquinones, saponin etc.

Chapter III: Methodology 1. Materials 3 mini cups aloe Vera gel 2 mini cups 70% Isopropyl Alcohol 3 teaspoon ripe papaya ( crushed) 2 teaspoon papaya seeds 2 teaspoon water Bowl Whisk Mortar and pestle Teaspoon

Sterile bottle Sterile gloves Funnel Cotton 2. Procedure a. Place 3 mini cups of aloe Vera gel and 2 mini cups of Isopropyl Alcohol in a bowl then thoroughly mix. b. Grind 3 teaspoons of ripe papaya pulp and 2 teaspoons of papaya seeds mixed with 2 teaspoons of water in a mortar and pestle then set aside. c. Mix the ingredients all together with continuous stirring then set aside for 1 day in an air tight sterile container. d. Filter the finish product and place it in an air tight sterile dispenser. Make two experiments. Experiment A as the controlled product (properly made according to procedure) and Experiment B as the experimental product (not stored in a tightly closed container), use papaya fruit extracts as the natural ingredient and compare the two experiments according to the following: a. viscosity b. color c. smell d. effectivity To test the effectivity of the products, test them by integrating them (with the aid of a medicine dropper) on glass slides contaminated with dirty hands. Observe each slide through the microscope and note if the bacteria will be killed. Compare also the result using commercialized hand sanitizers. Chapter IV : Data and Result VISCOSITY Viscous watery COLOR colorless colorless SMELL Papaya scent no scent EFFECTIVITY More effective effective

EXPERIMENT A EXPERIMENT B

The product in experiment A is more viscous than the product in experiment B. Product A retained its features from the time it was done. It was observed that the

unsealed experiment (Experiment B) became watery. The ingredients namely alcohol, aloe gel and scent evaporated thereby, leaving only the papaya extracts and water as the main content. CHAPTER V: CONCLUSION AND RECOMMENDATION: Ripe and unripe Carica Papaya extracts especially those from its epicarp, endocarp and seeds have antibacterial properties, that even if it is used singly or combined with other ingredients will still produce its antibacterial effect. The other ingredients is used to improve the consistency, appearance, and smell and served as a preservative to prolong the shelf life of the end product. We therefore recommend the use of Carica papaya as an antibacterial hand sanitizer because it does not only serve its antibacterial effect, but also a moisturizer and natural in nature thereby side effects of using it are lesser.