This document outlines a proposal for a study to test the antimitotic activity of an organic pesticide made from anahaw leaves. The study would involve extracting compounds from anahaw leaves and testing their ability to inhibit cell division in onion root tip cells. Specifically, the roots would be treated with different concentrations of the leaf extract and a control group. The number of dividing and non-dividing cells would then be counted under a microscope to calculate a mitotic index and determine if the extract has antimitotic properties. If shown to be effective against pests while being less harmful to the environment than synthetic pesticides, the anahaw extract could provide a safer alternative for agriculture.

1. LIKHA PROPOSAL
(1) PROJECT PROFILE
PROJECT TITLE: Homemade Organic Pesticide Against Pest
NAME OF PROJECT PROPONENT/S:
Untalan, Kwenn Leen Alhia R.
Dumlao, Auxyllume Zhynmagne P.
REGION:12 DIVISION: COTABATO
SCHOOL: MLANG NATIONAL HIGH SCHOOL
GRADE LEVEL: 9
PROJECT DURATION (Number of months): Five (5) Months
EMAIL: alhiauntalan@gmail.com CONTACT NUMBER: 09203326012
auxyllumed@gmail.com 09706973052
(2) CATEGORY OF RESEARCH
_______ PHYSICAL SCIENCE
____/___ LIFE SCIENCE
_______ ROBOTICS AND INTELLIGENT
MACHINES
_______ MATHEMATICS AND
COMPUTATIONAL SCIENCES
(4) THEME
_______ FOOD SAFETY
_______ WATER CONSERVATION
_______ CYBER SECURITY
_______ TRAFFIC/ROAD
CONGESTION
____ ___ HEALTH
_______ DISASTER MITIGATION
____/___ AGRICULTURE AND
ENVIRONMENT
_______ OTHERS (PLEASE
SPECIFY)
(3) CATEGORY
_____ INDIVIDUAL
___/___ TEAM
(5) EXECUTIVE SUMMARY
Mitosis is a process where a single cell divides into two identical daughter cells for
growth and to replace worn-out cells (Yourgenome, 2017). Errors in mitosis can cause abnormal
cell division, which can lead to the formation of tumors. Scientists trace 100% all cancer types
back to harmful mutations multiplied by mitosis. Traditional treatments and synthetic drugs for
cancer are costly and 25% of cancer patients cannot afford these treatments (Xiaochi, Hai Tao,
Liqun, & Lvguang, 2017). The leaves of Anahaw contain saponins, which is known to inhibit
tumor cell growth by cell cycle arrest, and apoptosis, also tannins, etc. The researchers decided
to conduct a study to test the antimitotic activity of anahaw. The antimitotic activity of anahaw

2. will be tested by immersing the roots of Allium cepa in three different amounts of extracts of
anahaw, distilled water as negative control, and Methotrexate as positive control. Three hours
later, the root tips will be observed under a microscope to record the number of non-dividing
and dividing cells (Shivasharanappa & Londonkar, 2014). The data will be gathered by
measuring the mitotic index.
(6) INTRODUCTION
(6.1) RATIONALE/SIGNIFICANCE
Pesticides cause short-term adverse health effect, called acute effect, as well as
chronic adverse effect that can occur month or years after exposure. Example of acute
health effects include stinging eye, rashes, blisters, blindness, nausea, dizziness,
diarrhea and death.The main purpose of pesticides is chemical developed to repel,
contrl or kill pest. Pest can be insects, weeds, fungi or rodents but pesticides can be
very dangerous to environment. That why there is a organic pesticides. Organic
pesticides have many advantage over synthetic ones and may be more cost-effective as
whole, considering the environmental cost of chemical alternatives. Organic pesticides
are biodegradable, barely leaves residues in the soil and are less likely to harm humans
or animals. The benefits of going organic and natural pesticides. By using organic you
reduce pollution and won’t introduce new harmful chemicals into soil, plants, and
ultimately our food. This study is expected to be beneficial to the following:
Agriculture. This will improve agriculture because organic pesticide is safe
and production for medical purposes will provide a safer and more environmentally
friendly for plants and animals.

3. Future Researchers. This research will contribute to other academics' collective
research and understanding. Other researchers may be able to expand on or draw
inspiration from this study to do further research.
Good Health ad Well Being. As a prevention of pest by controlling pests thereof,
it will help decrease the number of people with diseases caused be insects
(6.2) SCIENTIFIC BASIS/THEORETICAL FRAMEWORK/MATHEMATICAL
THEORY INVOLVED
The disruption of normal cell cycle regulation can result in illnesses like cancer.
When the cell cycle is uncontrolled, cells divide randomly and accumulate genetic
mistakes, which can result in cancerous tumors (MedlinePlus Genetics, 2017). Anahaw
leaves are usually used in landscaping, occasions, bouquets, awards, and medals
(matichub, 2021). It is also commonly found in the Philippines, especially in Luzon
(Benguet, La Union, Cagayan, Pangasinan, Zambales, Pampanga, Laguna, Ouezon,
Camarines, Albay), Negros, Cagayan de Oro and in the provinces of Mindanao
(Growing Anahaw, 2018). Anahaw leaves contain saponins, which are known to inhibit
tumor cell growth by cell cycle arrest and apoptosis, also tannins, etc (Dagalea et al.
2021). As such, the researcher decided to conduct a study to test the effectiveness of
the antimitotic activity of anahaw.
(6.3) OBJECTIVES
GENERAL: Generally, this study aims to determine the antimitotic activity of Organic
pesticide can be effective against pest and less harmful in environment.
SPECIFIC: Specifically, the study aims to:
1. To control pest
2. Determine if it is less harmful to environment.

4. 3. To lessen the disease caused by pest like malaria and dengue.
(7) REVIEW OF RELATED LITERATURE
(8) METHODOLOGY
Research Design
The researcher will implement Post-test only Control Group Design since this
type of experimental design uses variables randomly allocated to an experimental group
and a control group. The experimental group is exposed to a treatment, and both groups
are measured afterwards.
X O
Fig. 2 Schematic diagram of the research design
Collection of Anahaw Leaves
The researcher will be using 20 healthy anahaw leaves with size 1 meter in
diameter. The Anahaw leaves will be collected at Sto. Canibungan, Brgy. Daga, Cadiz
City. Leaves with scars and blemishes will not be collected.
Authentication of Anahaw Leaves
The leaves will be brought and verified at the Bureau of Plant Industry in
Bacolod City.
Preparation of Materials
The thorns of anahaw leaves will be removed then the leaves will be washed,
air-dried for 24 hours and cut into small pieces so that it can fit in a container to be
brought to the Negros Prawn Producers Cooperative Diagnostic and Analytical

5. Laboratory to be powderized for extraction. The researchers intends to conduct this study
at Negros Prawn Producers Cooperative Diagnostic and Analytical Laboratory.
Extraction of Anahaw Leaves
A hundred grams of powder of anahaw leaves will be macerated with ethanol (1g
: 5ml ethanol) in a sealed container for 5 days at room temperature with occasional
shaking. Extract will be filtered through Whatman No.1 filter paper and evaporated to
dryness under vacuum below 50ºC to obtain the extract. The extract obtained will be
kept at 4ºC for future use (Alamgir, & Rahman, 2013).
Quantification of Phytochemicals
Plants contain numerous chemical constituents that are responsible for eliciting
various physiological and therapeutic responses. Phytochemicals will be detected using
phytochemical screening in Negros Prawn Producers Cooperative Analytical and
Diagnostic Laboratory. The extracts of ethanol will be subjected to the various
qualitative tests for the detection of plant's phytochemicals like
alkaloids, glycosides, tannins, carbohydrates, coumarins, saponins, flavonoids, proteins
etc.
Antimitotic Assay in Allium cepa
The A. cepa root tip assay is the most common method for testing compounds
with antimitotic activity. The mitotic division in root tip cells resembles cell division in
normal human and cancer cells. Hence, onion root tip cells can be used for testing of
drugs with anticancer activity (Raheel, Saddiqe, Iram, & Azfal, 2017). Antimitotic
activity will be evaluated using the root of A. cepa roots. A. cepa bulbs will be sprouted

6. on wet stand at room temperature, when the roots will be about 2-3 cm long the
bulbs will be placed on vials containing the concentrations 100% S. rotundifolius
leaves ethanolic extract (10 ml), 75 % S. rotundifolius leaves ethanolic extract and 25%
distilled water (7.5 ml : 2.5 ml), 50% S. rotundifolius leaves ethanolic extract and 50%
distilled water (5 ml : 5 ml), 25% S. rotundifolius leaves ethanolic extract and 75%
distilled water (2.5 ml : 7.5 ml) such that the roots will be
immersed in the extracts. The duration of extract treatments for each bulb will be 3
hours. Three bulbs will be used for each extract. The sprouted roots will also be treated
with distilled water (Control group) and methotrexate (0.1mg/ml, Standard drug).
Microscopic Examination and Determination of Mitotic Index
The root tips (2–3 mm) will be cut and fixed in 45% acetic acid:1 N HCl (9:1).
The tips were transferred to 1 N HCl for softening of cell walls and after 5 min tips were
squashed and stained with 0.1% methylene blue. The researcher will observe the roots,
specifically the root apical meristem where cells are capable of repeated division,
approximately five hundred microns above the tip, under light microscope with 40x
magnification. In all cases cells showing different stages of mitosis were counted
(Raheel, Saddiqe, Iram, & Azfal, 2017).
Data Gathering
The researchers will be able to gather data by measuring the mitotic index using
the formula below taken from the article of Raheel, Saddiqe, Iram, and Azfal (2017):
Mitotic index
MI=P+M+A+T / Total cells, where P is prophase, M is metaphase, A

7. is anaphase and T is telophase.
Analysis of Data
The data will be analyzed using the Analysis of Variance (ANOVA) and
Pearson’s Correlation. ANOVA is used to assess potential differences in a scale-level
dependent variable by a nominal-level variable having 2 or more categories. ANOVA
will be used to determine if there is a significant difference between the varying
concentrations of S. rotundifolius ethanolic leaves extract, negative control, and positive
control in terms of mitotic index. The test statistic Pearson's correlation coefficient
assesses the statistical link, or association, between two continuous variables (Statistics
Solutions). Pearson’s correlation coefficient will be used to determine if there is a
significant relationship between the varying concentrations of S. rotundifolius and the
mitotic index of Allium cepa. In this experiment, Anahaw leaves extract will be used at
different levels.
Disposal Procedures
All non-hazardous waste will be disposed and segregated appropriately in a
garbage bin respectively. All liquid materials that will be used will be disposed in a
separate beaker. All glassware that will be used will be autoclaved. The flasks, beakers,
and test tubes will be washed with disinfectant liquid soap and will be sprayed with
pure H2O. The workplace will be cleaned with a Lysol solution and sterilized for 30
minutes with UV light.
(9) EXPECTED OUTPUTS AND POTENTIAL IMPACTS
The researcher expects that the anahaw (Saribus rotundifolius) ethanolic leaf
extract will have an antimitotic activity on the roots of allium cepas. Also, the

8. researcher aims to make anahaw as a potential alternative treatment for cancer and
tumor and prevention thereof. Thus, this study will have a great significance to the
following:
a. Agriculture - more cultivation and production of Anahaw for medical use
which can also benefit our local farmers for they will have another source
of income.
b. Field of Medicine - this will create an opportunity to study anahaw for future
research in the field of medicine particularly for the prevention of tumor and
cancer.
c. Future researchers - this study will give more information about anahaw that
could be useful for future research.
d. In the field of biotechnology- this study will provide information that will
be useful for the creation of natural medicines.
e. Good Health and Well-Being - This study falls under the 17 Sustainable
Development Goals (SDG) (United Nations, 2015), as a potential cancer
treatment and prevention thereof, it will slow down the increase of number
of people having cancer.
(10) WORKPLAN AND TARGET DELIVERABLES
Activity
I.Proposal
Conduct of background reading Brainstorming of ideas January 3-5
Preparation of proposed study Making research proposal January 7-12
Presentation of proposed study January 17
Activity
II.Pre- Implementation July 2022
Obtaining Laboratory Schedule

9. Canvassing of Materials Listing all materials that is
required in conducting
Collection of Anahaw leaves Will collect 30 Anahaw Leaves
Acquiring red onions 20 red onions
Activity
III. Implementation July –
September
2022
Extraction of Anahaw Leaves 25 ml of etract will be
extracted
Phytochemical screening of
anahaw
Phytochemicals will be
detected using phytochemical
screening
Antimitotic Assay in Allium
cepa
Bulbs will be placed on a wet
stands, 15 onion bulbs in total
and 15 wet stands
Activity
IV. Post-Implementation
Data Gathering and Analysis Measuring of mitotic index
Disposal procedure Waste will be thrown properly
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