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Amarillo and Ginger Extract as Mosquito-Killer

An Investigatory Project

Of

Jan Cedric M. Hernandez

Bansud National High School Regional Science High School Pag-asa, Bansud, Oriental Mindoro

Table of Contents I. Problem and its Settings a. Background of the Study b. General Objective c. Specific Objective d. Statement of the Problem e. Hypothesis f. Conceptual Framework g. Significance of the Study h. Scope and Limitations i. Definition of Terms II. Review of Related Literature and Study a. Review of Related Literature b. Review of Related Study III. Materials and Methods a. Research Method b. Materials c. Equipment/Apparatus and Utensils

d. Procedure e. Training of the Panelist f. Evaluation of the Products g. Statistical Treatment IV. Bibliography

Chapter I PROBLEM AND ITS SETTINGS Background of the Study Nowadays, number of skin diseases was being diagnosed by scientists during these days such as malaria, dengue and yellow fever. These were caused by the increasing population of disease-carrying mosquitoes which was spread all over the world especially in our country. A mosquito bite can cause severe irritation through an allergic reaction to the mosquitos saliva. Many had died because of the mentioned disease. Most malaria cases in the country or even in the world occur in rural areas and especially in forested, swampy, hilly and mountainous regions. The victims of the said disease were the poor people and some are the indigenous. Many studies have been aiming only to repel mosquitoes but repellant drives mosquitoes away only for a short time. Aside from eliminating their breeding places, one solution to the mentioned problems above is to kill mosquitoes especially those disease-carrying mosquitoes in our home and backyard. These are some compounds that can kill mosquitoes. In order to help reduce the spread of mosquitoes, the researcher made an investigatory project that aims to produce a mosquito-killer. It was made readily from available natural ingredients with easy procedure so that people especially in rural areas could help decrease the population of mosquitoes in their place. Through the use of amarillo and ginger extract, diseasecarrying mosquitoes will die through inexpensive materials, without much effort and with more comfortable.

General Objectives This study determined the effectiveness of amarillo and ginger extract as mosquito-killer. Specific Objectives Specifically , this study aimed to: 1. Determine the active components of ginger and amarillo that might kill mosquitoes 2. Know the significant difference between the experimental product amarillo and ginger extract and the commercial product in terms of: 2.1. Number of mosquitoes killed 2.2. Odor 3. Evaluate if the of amarillo and ginger extract, when boiled will release a chemical that can kill mosquitoes. 4. Make a mosquito-killer biologically and naturally. 5. Lessen the number of cases of disease cause by insects. 6. Determine the effect of ginger and amarillo in killing mosquitoes. 7. Know if the said mosquito-killer is cheaper than the commercial ones. 8. Determine if the experimental product can affect the health or can kill the person who accidentally took it.

Statement of the Problem This research study aims to produce a substance that can kill mosquitoes.

Specifically this research investigation intends to answer the following questions: 1. What is the active components of ginger and amarillo that might kill mosquitoes? 2. Is there significant difference between the experimental product amarillo and ginger extract and the commercial product in terms of: 2.1. Number of mosquitoes killed? 2.2. Odor 3. Can the of amarillo and ginger extract, when boiled will release a chemical that can kill mosquitoes? 4. Can the researcher make a mosquito-killer biologically and naturally? 5. Can the researcher lessen the number of cases of disease cause by insects? 6. How effective is the ginger and amarillo extract in killing mosquitoes? 7. Does the said mosquito-killer is cheaper than the commercial ones? 8. Will this experimental product can affect the health or can kill the person who accidentally took it?

Statement of Hypothesis In the study Amarillo and Ginger Extracts as Mosquito-Killer, the following was hypothesized: 1. Boiling amarillo and ginger extract in water, can kill and decrease the number of mosquitoes.

2. There is significant difference between the experimental product amarillo and ginger extract and the commercial product in terms of: 2.1. Number of mosquitoes killed 2.2. Odor 3. Side effects can occur if one accidentally takes the amarillo and ginger extract. 4. The experimental product amarillo and ginger extract is cheaper than the commercial ones because the materials used to produce that experimental product is cheap and are abundant in the Philippines and other tropical countries. Conceptual Framework

Independent Variables

Dependent Variable
Formulation of the product Amount of heat Time of application Commercial Product Volume of the extracts Number of mosquitoes killed Odor

Intervening Variable Temperature of the Surroundings State of the Environment

Significance of the Study Mosquito diseases are one of the major health problems in the world especially in the Philippines. Most people think that protecting one family from harm is expensive and difficult to achieve. Some knows that it need many high-quality mosquito repellant and mosquito-killer to kill and control the number of mosquitoes but the real is we can achieve is just how one resourcefully utilizes the cheap, the natural abundance of nature and the untiring quest to alternatives and remedies in killing mosquitoes. This study will promote awareness in lessening the number of mosquitoes all over the world especially in the Philippines without spending much money. Specifically this will bring benefits to the following: Government: This would provide the government a new avenue in lessening the number of mosquitoes. The materials that will be used are organic in which its availability is easily utilized and therefore they will spend less amount of money in the treatment for the abundance of mosquitoes in the society Residents/People This study raises the awareness level of residents in our country especially the awareness from the diseases that they may encounter. They would also be able to foresee the importance of being resourceful of the things in our surroundings.

Babies/Children This would provide these children and babies protection from diseases carried by mosquitoes such as malaria and dengue. The materials of these are organic so these childrens health is not affected. Squatters This study would provide squatters from diseases such as malaria, dengue and yellow fever that they may encounter due to their surroundings. They can save more money than paying money if they might get sick. They can sleep earlier and more comfortable, too because there is less mosquito that might bite them. Natives This study introduces the new easy way in lessening the number of mosquitoes in place where almost most natives live like in mountains. This study could raise their awareness in the disease that they might have encounter and they could know how to prevent it by using or making amarillo and ginger extracts. Researcher This study provide the most information than other person that will read this research. This will increase the awareness and experienced in life of the researcher. After this research was proved, the researcher can make of amarillo and ginger extract anywhere, and the researcher can save some money. This researcher can benefit in this research the best.

Scope and Limitations This science investigatory project was focused in identifying the effect of amarillo and ginger extract as mosquito-killer. Three experimental formulations were prepared. The product was tested on mosquitoes, and its rate of killing, time after the product to effect, number of mosquitoes and odor were observed. Moreover the significant differences among the three formulations and the control group were measured. The aspects looked into were the availability and effectivity of the ingredients to be used in the experiment, the methods and strategies and proposed solution to mosquito-killer industry problems. The experimentation was conducted from September 2012 to March 2013 at Alcadesma, Bansud, Oriental Mindoro. Data processing and analysis were done in Learning Resource Center Laboratory of Bansud National High School-Regional Science High School for Region IVMIMAROPA. Other variable like intervening such as temperature and state of the environment were beyond in the scope. Definition of Terms The following terms are operationally defined: Amarillo. This refers to a rather course, erect, glabrous branched, rank-smelling annual herb, 0.4 to 1 centimeters high. Leaves were 4 to 7 centimeters long, deeply pinnatified with linear-lanceolate segments (Stuart & Santiago, 2012). In this research

study, the word amarillo was defined as the plant that was in the study that provides the extracts needed to kill mosquitoes. In this study it is also called marigold and ahito. Formulations. This term refers to a substance prepared according to a formula (Memidex, 2013). As used in the study, formulations refer to the quantity of two liquid objects prepared as one. In this study, there are 3 formulations and these formulations were often called as treatment. These refers to Treatment 1, Treatment 2 and Treatment 3. Mosquito. This term was defines as any numerous dipterous insects of the family Culicidae; the females which suck the blood of animals and humans; some species transmit certain diseases like malaria and yellow fever (Farlex Inc., 2013). In this study, mosquito refers to the type of the insect that will be suspected to be killed by the extracts of amarillo and ginger. Organic. This was refer a of pertaining to, or of the nature of animals and plants (Websters Comprehensively Dictionary, 1958). In this study, this word mean not using any kind of chemical but instead using only ingredients that can be found in nature. In this study, these determined the amarillo and ginger. Temperature. This refers to the condition as regard heat or cold(Websters Comprehensively Dictionary, 1958 ). In this study, it refest to the degree of coldness and hotness of the environment.

Chapter II REVIEW OF RELATED LITERATURE, STUDIES AND LEGAL BASES

Review of Related Literature In addition to being looking attractive of marigold, the flowers are also edible, and have natural pesticidal properties that allow them to resist insect infestation. Some gardeners take advantage of this property to garden organically, protecting more sensitive species with a barrier of this plants. Any plant in the genus Tagetes can be considered a marigold, along with several other blooming plants in the Asteraceae family. (Smith & Harris, 2013) It is significant to the researchers study for the reason that it gives more information to the background of the marigold. Like the information given that marigold has natural pesticidal properties, through that the researcher can hypothesize that the study doing by the researcher can be effective or was truly true. According to Tvedten , marigolds are whole plant extracts of this species of marigold are extremely toxic to adults and larvae of mosquitoes, e.g., Aedes aegypti (L.). Caution: The extracts may also kill or negatively effect non-target aquatic insects and other organisms. (Stephen Tvedten) It is significant to the researchers study for the reason that it can warn the researcher or people that may do the experimentation for marigold that can gave them risk or danger. The researcher and the reader can be convinced more through the information.

According to Santiago and Stuart in 2012, there are studies that proved that ahito or marigold had many benefits or uses. The following are the effects that marigold can benefit to us in different topics: Larvicidal: Study of T patula essential oil on fourth instar larvae of mosquito species showed greatest activity against A aegypti, followed by An. stephansi and C quinquefasciatus. Results were compared to synthetic insecticide, malathion. Insecticidal: Study of crude extracts and fractions of flower of T erecta was tested for insecticidal activity against stored product insect pest, Trilobium castaneum. The chloroform extract showed highest toxicity against both larvae and adults of T castaneum. It is significant to the researchers study for the reason that it gives information what type of mosquito can the product of the study can kill. It also gives what amarillo or marigold can kill.

According to Khazdesignz (2004), ginger contains 0.25 - 3.3% volatile oil, pungent principles (gingerols and shogaols), about 6 - 8% lipids composed of triglycerides, phosphatidic acid, lecithins, free fatty acids (lamic, palmitic, stearic, oleic, linoleic, etc.), proteins (about 9%), starch (up to 50%), vitamins (especially niacin and A), minerals, amino acids, resins, among others.

Studies conducted in lab animals found that both gingerols and shogaols have analgesic (shogaols), sedative, anti-pyretic, anti-bacterial and gastro-intestinal motility effects. Other investigations show [6]-gingerol to have anti-oxidative, anti-inflammatory and anti-tumour

promotional activities.9, 11 The anti-inflammatory properties of gingerols have been linked to the structure, which is said to be similar to capsaicin, a known pain killer/reliever. Gingerols have been found to inhibit the enzyme cyclooxygenase (COX) that causes inflammation.I2 [6]shogaol was also found to exhibit anti-inflammatory activities. Ginger has also been found to reduce cholesterol levels by promoting its excretion and impairing its absorption. It was indicated ~hat this cholagogic effect (If ginger was due mainly to [6]- and [10]- gingerols.9, 13 Gingerols have been reported to have potent cardiotonic and thermogenic activities. It is significant to the researchers study for the reason that it can give the researcher and the reader prior knowledge about what is a ginger, what are its active properties and what its other function. It also gives the major component of ginger that can help the researcher in determining the component of ginger that can kill mosquitoes. Review of Related Studies According to Puengphian and Sirichote reseach (2007), ginger is one of the most widely used herbs that contains several interesting bioactive constituents and possesses health promoting properties. [6]-Gingerol, a major pungent ingredient of ginger, also has potent antioxidant activity. Monitoring of [6]-gingerol content during the drying process, ginger extraction with supercritical CO2 and bioactive properties analysis of extracts were performed. Fresh mature ginger rhizomes with 94.17 0.16% moisture content were dried using a rotary air dryer at 55 2C for 11 hours to achieve a moisture content of 11.54 0.29%. After the drying process, [6]gingerol content of the ginger rhizomes were reduced from 21.15 0.13 to18.81 0.15 mg/g dry weight basis. Dried ginger was then pulverized to coarse powder, approximately 0.5 mm in diameter, prior to extraction. The supercritical CO2 extraction of ginger was undertaken with

two conditions of 200 bar at 35C and 230 bar at 40C. The results showed that the extracts from both conditions had [6]-gingerol contents of 238.94 0.79 and 170.50 0.45 mg/g extract, total phenolic contents of 183.96 1.25 and 126.04 0.72 mg gallic acid/g extract, respectively. In addition, the ginger extracts showed antioxidant activities using DPPH (1,1-Diphenyl-2picrylhydrazyl) radical scavenging assay, compared with BHT standard, expressed as EC50, that were 13.09 1.77 and 26.68 1.76 g/ml, respectively. The antioxidant activities using ABTS (2,2-azinobis [3-ethylbenzothiazoline-6-sulfonic acid]) radical cation scavenging assay were 813.33 6.67 and 724.44 7.70 mol Trolox/g extract,respectively. It is significant to the researchers study for the reason that through this information, the readers can have additional knowledge about some components of the amarillo and ginger extract. In Babus study (2007), essential oil of Tagetes minuta, a kind of amarillo was distilled in vacuo in a mini distillation apparatus to study and compare the qualitative and quantitative characteristics vis--vis oil distilled by conventional distillation technique at normal temperature and pressure (NTP), which revealed that higher oil yield (1.56%) was obtained in the oil distilled by conventional distillation technique at NTP and decreased gradually with increase in vacuum to 150 mmHg (1.16%), 200 mmHg (1.07%) and 225 mmHg (0.91%). Concentration of monoterpene hydrocarbons (MH) was found maximum (55.3%) in the oil distilled at NTP and decreased with increase in vacuum to 150 mmHg (35.2%), 200 mmHg (31.4%) and 225 mmHg (27.5%). On the contrary, oxygenated monoterpenes (OM) were found minimum (22.9%) in the NTP distilled oil but increased with increase in vacuum to 150 mmHg (55.4%), 200 mmHg (61.3%) and 225 mmHg (65.3%). Sequential evolution of chemical compounds was also studied by distilling the oil at 225 mmHg vacuum and found that the lipophilic effect plays an important

role in distillation of MHs. Due to the high concentration of these unsaturated MHs, the oil gets polymerized easily when exposed to unfavorable conditions and this can be minimized by reducing the concentration of these compounds by distilling the plant material under vacuum thus increasing the shelf life of the oil. The physicochemical properties of the oil were also evaluated. Vacuum distilled oil samples possessed better solubility in alcohol than NTP distilled oils. The olfactory evaluation of the samples revealed that the oil distilled at NTP possessed powerful, peculiar and disagreeable odour with burnt note, whereas the oils distilled in vacuo has pleasant odour varied from powerful fruity odour without any burning note to near true-to-nature odour of the flowers with increase in vacuum. Due to these improved characteristics, the in vacuo distilled oil of T. minuta is recommended for exploring its application in high grade perfumes, flavours and other formulations. It is significant to the researchers study for the reason that it gives another idea aside from making extracts of amarillo and ginger is, we can also make it in the form of oil and through the ideas and knowledge contribute by that review of related study, we do not have to experiment what formulation is the best for the product. In Peng, et al. study (2012), twenty-nine phenolic compounds were isolated from the root bark of fresh (Yunnan) ginger and their structures fully characterized. Selected compounds were divided into structural categories and twelve compounds subjected to in-vitro assays including DPPH radical scavenging, xanthine-oxidase inhibition, monoamine oxidase inhibition, rat-brain homogenate lipid peroxidation, and rat pheochromocytoma PC12 cell and primary liver cell viability to determine their antioxidant and cytoprotective properties. Isolated compounds were also tested against nine human tumor cell lines to characterize anticancer potency. Several diarylheptanoids and epoxidic diarylheptanoids were effective DPPH radical scavengers and

moderately effective at inhibiting xanthine oxidase. An enonedione analog of 6-shogaol (compound 2) was isolated and identified to be most effective at protecting PC12 cells from H2O2-induced damage. Almost all tested compounds inhibited lipid peroxidation. Three compounds, 6-shogaol, 10-gingerol and an enone-diarylheptanoid analog of curcumin (compound 6) were identified to be cytotoxic in cell lines tested, with KB and HL60 cells most susceptible to 6-shogaol and the curcumin analog with IC50 < 10 M. QSAR analysis revealed cytotoxicity was related to compound lipophilicity and chemical reactivity. In conclusion, we observed distinct compounds in fresh ginger to have biological activities relevant in diseases associated with reactive oxygen species. It is significant to the researchers study for the reason that it proves that ginger has the active component being called 6-shogaol or simple shogaol. It can give better and more ideas about ginger and what it is composed of.

Chapter III MATERIALS AND METHODS Research Method Experimental method was used in the study, particularly, parallel-group design. This design was must appropriate because three or more groups were used at the same time with only single variable was manipulated or changed. The experimental group varied while the parallel group served as control for comparative purposes. In this study, there were three formulations, treatment 1 (T1), treatment 2 (T2), treatment 3 (T3) and a control group. Treatment 1 was made up 25 mL of amarillo flower extract and 25 mL of ginger extract. Treatment 2 was made up 30 mL of amarillo flower extract and 20 mL of ginger extract. Treatment 3 was made up 20 mL of amarillo flower extract and 30 mL of ginger extract. Materials The materials used in the study were kg peeled ginger root, kg amarillo flower and water, about 500 mL. Equipment and Utensils The equipment and utensils used in the preparation of amarillo and ginger extract were mortar, pestle, 2 clean plastic containers, grater, tripod, alcohol lamp, match and evaporating dish.

Procedure In order to do the amarillo and ginger extract, gathering and preparing the materials and equipment were the first thing that the researcher had done. The researcher divided the whole procedure into two: the pounding process and the boiling process. The first things in the first process done were to get the grater and kg peeled ginger root. Using the grater, the researcher grated all the peeled ginger root until all were finished, put it in a plastic container and covered it with its lid. Then the researcher used the mortar and pestle to pound the kg of amarillo flowers; added continuously the pieces of amarillo flower in the mortar; small amount in the first time and then when it was more compact to each other, the researcher added again some amarillo flower and about 2.5 mL water. After all the amarillo flowers was pounded, the researcher placed it into another container, and also place its lid. Then wash the mortar and pestle and let it dry. After pounding the amarillo flowers, the researcher got the ginger and placed it into the mortar and had done the same process in pounding the amarillo. After it was done, the researcher placed the amarillo extracts and ginger extracts in the evaporating dish according to the formulations or treatment found in the research method. Then the researcher had done the same set-up in the picture below for the boiling process.

Evaporating Dish

Tripod Alcohol Lamp

Figure 1: Set-up for the Boiling Process

After preparing the set-up, the researcher lighted the candle or alcohol lamp using matchstick. Lastly the researcher let the extracts evaporate. Training of the Panelist The panelists were chosen by picking randomly from the whole population fron the different barangays of Bansud, Oriental Mindoro. After choosing the panelist, the researcher trained them. The only attribute that the panelist will evaluate is the products odor, the researcher explained that the panelists should rate all the products by smelling through it but before that the researcher asked them that the products that they will evaluate is placed into resealable plastics, labeled A,B, C and D. The researcher explained briefly that in testing the product, the panelists should rate each product according to the likert scale presented in the Evaluation of the Product that 5 is the highest score the product can have in each attribute and 1 is the lowest. The researcher also explained that the researcher will provide the extracts in testing the products. Evaluation of the Products The trained panelists were asked to evaluate the acceptability of the different attributes of the products using a 5-point Likert scale. Generally, the acceptability was also asked.

Odor In order to test whether there is a significant different between the experimental products and commercial product, the researcher conducted or survey with 30 respondents by smelling the product and signing a rating scale 1 to 5 based on the following scale:

Scale 5 4 3 2 1

Odor Pleasant Odor Odorless Foul Odor Slightly Foul Odor Strong Foul Odor

Table 1: Scale According to Odor Effectiveness In order to test the effectiveness of the product, the effect of the products according to the number of mosquitoes killed, were counted and recorded in the table. Statistical Treatment The statistical techniques used in the analyzing and interpreting the data were weighted arithmetic mean and the F-test or two-way analysis of variance (ANOVA). Mean was used to

determine the acceptability of the quantity attributes of the three formulations and the experimental group. The formula is as follows:

where stands for the weighted arithmetic mean; f, frequency; x, weight; , sum of the product of f and x; and , sum of the frequency. F-test or two-way ANOVA was used to determine the significant difference on the acceptability of the quantity attributes of the three formulations and the experimental group. The formula is as follows:

where for error.

stands for F-value for samples;

, mean square for samples; and

, mean square

where for error.

stands for F-value for panelists;

, mean square for panelists; and

, mean square

In this connection, the researcher opted for 5 percent level of probability to determine the degree of significance of the findings with 99 percent reliability.

Bibliography

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