Group 1 Research Study 2024 Final Chapters 1 3 Feb 20 2024 1

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NEW LUCENA NATIONAL COMPREHENSIVE HIGH SCHOOL

SPECIAL CURRICULAR PROGRAM IN SCIENCE, TECHNOLOGY AND ENGINEERING
Brgy. Poblacion, New Lucena, Iloilo
CHAPTER 1
INTRODUCTION
1.1 The Problem and its Background
The Allium cepa assay is an efficient test for chemical screening and in situ
monitoring for genotoxicity of environmental contaminants. The test has been used
widely to study genotoxicity of many pesticides, revealing that these compounds can
induce chromosomal aberrations in root meristems of Allium cepa.
Allium cepa, commonly known as red onion is a perennial herb with the stem in the
underground bulb. Onions belong to the Liliace Family. It has a high dietary fiber and
sugar content of about 90% of water. A diet rich in vegetables has been beneficial in
preventing of the more prevalent and relevant diseases nowadays.
Allium cepa root tip bioassay is considered as a simple and reliable test model to
assess the genotoxic potential of chemicals due to its characteristics proliferation rate,
high percent of dividing cells and relatively prominent cells with lesser number of
large sized monocentric chromosomes (2n=16) that stain well (Fiskejo, 1997).
Furthermore, they have easily distinguishable genetic endpoints, such as
chromosomal aberrations (CAs), changes in ploidy and sister chromatid exchanges
(Kumari et al., 2011). CAs have been determined by using a cepa test since 1920,
(Sataphaty and Swammy, 2015).
Tawa-tawa herb or scientifically known as Euphorbia hirta linn plant, common
hairy herb in the Philippines is a pantropical weed. It is a feeder that grows in opened
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grasslands, roadsides and pathways. It is widely used in traditional herbal medicines
across many cultures, particularly for asthma, skin ailment and hypertension. It is
commonly used as treatment for dengue. Scientist studies have shown that tawa-tawa
is rich in bioactive compounds like phenolic and flavonoids which may be
responsible for its anti-dengue properties.
The toxicity of tawa-tawa (Euphorbia hirta linn) boiled crude extract will be
evaluated using Allium cepa root tip test to determine the chromosomes will be
destroyed after the living organisms will consume it continuously. Especially from
this present day, some medical professionals has declared that the tawa-tawa herb
(Euphorbia hirta linn) can affect in reducing the dengue properties.
1.2 Statement of the Problem
The main purpose of the study is to evaluate the toxicity of Tawa_tawa
(Euphorbia hirta linn) on Red onion (Allium cepa) root tip.
1. What is the mean of Red Onion (Allium cepa) root lengths when exposed
with the negative control (distilled water), 100% and 50% concentration of
Tawa-Tawa (Euphorbia hirta) extract?
2. What is the mitotic index of Red Onion (Allium cepa) root tip chromosomes
when exposed with the negative control (distilled water), 100% and 50%
concentration of Tawa-Tawa (Euphorbia hirta) extract?

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3. Is there a significant difference between the root length mean of Red Onion
(Allium cepa) root tip when exposed with the negative control (distilled
water), 100% and 50% concentration of Tawa-Tawa (Euphorbia hirta)
extract?
4. Is there a significant difference between the mitotic index of Red Onion
(Allium cepa) root tip when exposed with the negative control (distilled
water), 100% and 50% concentration of Tawa-Tawa (Euphorbia hirta)
extract?
1.3 Hypothesize
1. There is no significant difference between the root length mean of Red Onion
(Allium cepa) root tip when exposed with the negative control (distilled water), 100% and
50% concentration of Tawa-Tawa (Euphorbia hirta) extract.
2. There is no significant difference between the mitotic index of R ed Onion
(Allium cepa) root tip when exposed with the negative control (distilled water), 100% and
50% concentration of Tawa-Tawa (Euphorbia hirta) extract.

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1.4 Conceptual Framework
Independent Variable Dependent Variable
Tawa-tawa (Euphorbia hirta):
50% Tawa-tawa herb extract + 50%

water Root length mean
100% Tawa-tawa herb extract Mitotic Index
Negative control (distilled water)
Figure 1. Paradigm shows the relationship of Tawa-tawa (Euphorbia hirta) extract and
its negative control (distilled water) as the independent variable and root length mean and
mitotic index as the dependent variable.
1.5 Significance of the Study:
The result of this study may hopefully benefit the following content:
1. Community. This study provides further knowledge in agricultural practices or
ecological considerations in community, potentially influencing on how Tawa-tawa
(Euphorbia hirta) is managed or used in certain environments.

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2. Dengue Patients. The result of this study provides more information and
knowledge on how does the Tawa-tawa extract affects the human body especially the
body organs.
3. Medical Professionals. This study provides knowledge on what is the toxicity
rate of the Tawa-tawa (Euphorbia hirta) toxicity on the human body especially to the
body organs.
4. DOST. They can use this study as their reference and guide for them to arrive at
supporting results and findings on how does the Tawa-tawa (Euphorbia hirta) herb
affects the health of the humans.
5. Future Researchers. This study is beneficial to the future researchers as this may
provide additional literatures involving studies about toxicity of Tawa-tawa (Euphorbia
hirta). It can provide important inputs to inform and develop parallel studies for further
research about related matters involving toxicity of Tawa-tawa (Euphorbia hirta).
1.6 Scope and Delimitation
This study is limited on determining the toxicity of Tawa-tawa (Euphorbia hirta)
extract on Red onion (Allium cepa) root tip chromosomes. This study will be conducted
at Poblacion, New Lucena, Iloilo. The amount of Tawa-tawa will be determined in the
study, the concentration of said treatment is either 50%, and 100% of Tawa-tawa
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(Euphorbia hirta) extract, as well as the negative control (distilled water) on Red onion
(Allium cepa) root tip.
Furthermore, the effect of the said treatments will be verified using the
measurement of root length mean and testing of mitotic index. There will be 3 trials with
3 replicates each treatment. The result of the root length mean and mitotic index will be
included in the statistical analysis.
1.7 Definition of Terms
1. Tawa-tawa (Euphorbia hirta), is abundant in open grasslands and waste
places. It is classified as a weed and is a native of India. This is widely distributed in the
Philippines, from sea level to an altitude of 500 meters. The leaves are hairy and elliptical
with slender dentate margin. The chemical constituents of this plant are the following:
gallic acid, quercetin and a phenolic substance C28H18O15. These substances are
responsible for its medicinal properties. (Wall, 2021)
2. Red Onion (Allium cepa) is highly valued for its therapeutic properties. It has
been used as a food remedy from time immemorial. Research shows that onions may help
guard against many chronic diseases. The purple skinned onion tastes great. Additionally,
it has several health benefits and is part of many home remedies and beauty solutions.
(Essa et al., 2016)

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CHAPTER 2
REVIEW RELATED LITERATURE
2.1 Related Literature
2.1.1 Allium cepa
There are number of toxic chemicals in the environment, they are mostly
discharged by industries into water, air and soil. The continuous use of chemicals, led the
world to establish various chemicals industries. The chemicals enter in our environment
through both natural and anthropogenic ways. Once they enter in our biological process,
it’s really difficult to eliminate them from the environment and disturb various
biochemical processes, leading to fatal results. Numerous potentially mutagenic
chemicals have been studied because they can cause mutagenic, damaging and inheritable
changes in the genetic material. Many thousands of toxic chemicals including
pharmaceuticals products, domestic and industrial wastes, pesticides and petroleum
products are present in the environment and new chemicals are being introduced every
year. No doubt, rapid progress of chemical industry has provided economic and social
benefits but at the same time it has accentuated the environmental and social problems.
Environmental biologists are presently concerned to safeguard the human beings from
exposure to chemicals. (Devi et al., 2020)

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Genotoxicity is to determine the magnitude of gen etic risk to man by an
environmental agents/ chemical under a specified level of exposure. Unfortunately, the
direct assessment in human is not feasible because of ethnic, logistic and practical
considerations. (Detels et al., 2017)
Even the epidemiological approaches used to detect genotoxic and carcinogenic
chemicals have limitations because detection is possible systems. There are many
employing wide variety of organisms ranging from viruses, bacteria, plants and insects to
human cell cultures and intact mammals to evaluate the mutagenicity of environmental
chemicals. In order to identify the harmful effects of substances in different
concentrations and time of exposure, a variety of tests have been employed, such as
cytogenetic tests. These tests are commonly used for biomonitoring the extent of
pollution and to evaluate the effects of toxic and mutagenic substances in the natural
environment. (Nations & Organization, 2019)
Higher plants constitute an important material for genetic tests to monitor
environmental pollutants. However this feature is due to the possibility of assessing
several genetic endpoints range from point mutation to chromosomal aberrations in cells.
Among the higher plant species, the most frequent ones used to evaluate
environmental contamination are Allium cepa, Vicia faba, Zea mays, Tradescantia,
Nicotiana tabacum, Crepis capillaris and Hordeum vulgare. But, still among these
species, Allium cepa (Onion) has been considered an efficient test organism to indicate
the presence of mutagnic chemicals due to its kinetic characteristic of proliferation and
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chromosome suitable for this type of study. A. cepa root chromosomal aberration assay
was described as an efficient test system routinely used to evaluate the genotoxic
potential of chemicals in the environment, due to its sensitivity and good correlation with
mammalian test systems. Thus A. cepa is an efficient test organism for environmental
monitoring, especially in contaminated aquatic environments. (Khan et al., 2021)
2.1.2 Euphorbia hirta
E. hirta, commonly known as tawa-tawa, is a small annual herb common to
tropical countries like the Philippines.1-4 The exraction of the whole plant is used as a
folkloric herbal treatment for dengue hemorrhagic fever in the country.5 A exraction
from the whole plant is given to dengue patients, 1 to 1.5 glasses every hour for 24
hours.6The plant is believed to have platelet increasing property. Different animal studies
showed consistent findings of a significant increase in platelet count.5,7-8 However,
health authorities warn the general public in using this plant for dengue management
because of insufficient efficacy and safety data available to support its clinical use and to
validate the claims and testimonials from patients or healthcare professionals who were
said to experience the clinical benefits of this plant. (Egbuna et al., 2021)
Herbal medicines should be evaluated in terms of quality, efficacy and safety.
While efforts are ongoing to determine the role of E. hirta for dengue, this study focused
on its safety. In particular, the acute oral toxicity of the crude aqueous extract of the
whole plant of E. hirta was determined in this study. The results of this preliminary study
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will then become a part of a database that is essential for the safety assessment of this
plant.
Tawa-tawa is usually abundant throughout the Philippines in waste places and
open grasslands. The plant is an annual hairy herb, usually much-branched from the base.
These branches are simple or forked, ascending or spreading up to 40 centimeters long
and often reddish or purplish. The leaves are opposite, distichous, oblong-elliptic to
oblong-lanceolate, 1 to 2.5 centimeters long, toothed at the margin, and usually botched
with purple in the middle. The plant deserves special attention because of its medicinal
properties. Local tradition credits that this plant can help pa-tients stricken with dengue
fever. While it does not fight with the virus, it promotes the develop-ment of blood
platelets and softens the effect of the viruses which attack the blood. However the
department of health advises the public not to be much dependent on the said herbal
medicine, despite its proven efficacy. This study was conducted to elucidate the structure,
antimicrobial and cytotoxicity of the extract. Twenty grams of air-dried tawa-tawa were
percolated in ethyl acetate of 200 ml for one week, rota-evaporated and purified by wet
column chromatography. The phyto-chemical tests revealed the presence of alkaloids and
tannins. The structure of the constituents was partially elucidated by ultraviolet spectra
and gas chromatography-mass spectra analysis. The λmax was at 420 nm with an
absorbance of 1.233 and GC-mass spectra characterized 100 compounds. The
antimicrobial analysis gave positive inhibition of P. aeruginosa, S. aureus, C. albicans
and T. mentagrophytes with activity index of 0.2, 0.3, 0.4 and 0.2 respectively. The
cytotoxicity test showed that a concentration of 36.7185 grams in water of 350 ml of the
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leaf extract was toxic based on the t-test of the live and dead cells by in-vitro analysis on
the lymphocytes from normal blood cells. (Hanif et al., 2019)
2.2 Related Studies
2.2.1 Allium cepa
Higher plants are recognized as excellent genetic models to detect environmental
mutagens and are frequently used in monitoring studies. Among the plant species, Alium cepa has
been used to evaluate DNA damages, such as chromosome aberrations and disturbances in the
mitotic cycle. Employing the A. cepa as a test system to detect mutagens dates back to the 40s. It
has been used to this day to assess a great number of chemical agents, which contributes to its
increasing application in environmental monitoring. The A. cepa is characterized as a low cost
test. It is easily handled and has advantages over other short-term tests that require previous
preparations of tested samples, as well as the addition of exogenous metabolic system. Higher
plants, even showing low concentrations of oxidase enzymes and a limitation in the substrate
specification in relation to other organism groups, present consistent results that may serve as a
warning to other biological systems, since the target is DNA, common to all organisms. The A.
cepa test also enables the evaluation of different endpoints. Among the endpoints, chromosome
aberrations have been the most used one to detect genotoxicity along the years. The mitotic index
and some nuclear abnormalities are used to evaluate citotoxicity and analyze micronucleus to
verify mutagenicity of different chemicals. Moreover, the A. cepa test system provides important
information to evaluate action mechanisms of an agent about its effects on the genetic material
(clastogenic and/or aneugenic effects). In the face of all the advantages that the A. cepa test
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system offers, it has been widely used to assess the impacts caused by xenobiotics, characterizing
an important tool for environmental monitoring studies, where satisfactory results have been
reported. (Jorgensen & Fath, 2022)
Onions are a very common and rich source of dietary flavonoids, and contain
three diverse and highly valuable phytochemicals in perfect proportion: flavonoids,
fructans, and organosulfur compounds. Phytochemicals have the potential to promote
health benefits in humans and offer protection from a variety of diseases, including
cancer. Apart from the main sulfur and flavonoid constituents, several other constituents
of intact onion, such as lectins (the most abundant proteins in onion), prostaglandins,
fructan, pectin, adenosine, vitamins B1, B2, B6, C, and E, biotin, nicotinic acid, fatty
acids, glycolipids, phospholipids, and essential amino acids, have been studied for their
biological effects over several decades. Several epidemiologic studies have reported that
the antiplatelet activity of onions is considered to be a property of organosulfur
compounds. In particular, a class of α-sulfinyl-disulfides (cepaenes), found in onion
extracts, has demonstrated antithrombotic activity. (Galanakis, 2021)
2.2.2 Euphorbia hirta
Tawa-tawa is usually abundant throughout the Philippines in waste places and
open grasslands. The plant is an annual hairy herb, usually much-branched from the base.
These branches are sim-ple or forked, ascending or spreading up to 40 centimeters long
and often reddish or purplish. The leaves are opposite, distichous, oblong-elliptic to
13
oblong-lanceolate, 1 to 2.5 centimeters long, toothed at the margin, and usually botched
with purple in the middle. The plant deserves special attention because of its medicinal
properties. Local tradition credits that this plant can help pa-tients stricken with dengue
fever. While it does not fight with the virus, it promotes the develop-ment of blood
platelets and softens the effect of the viruses which attack the blood. However the
department of health advises the public not to be much dependent on the said herbal
medicine, despite its proven efficacy. This study was conducted to elucidate the structure,
antimicrobial and cytotoxicity of the extract. Twenty grams of air-dried tawa-tawa were
percolated in ethyl acetate of 200 ml for one week, rota-evaporated and purified by wet
column chromatography. The phyto-chemical tests revealed the presence of alkaloids and
tannins. The structure of the constituents was partially elucidated by ultraviolet spectra
and gas chromatography-mass spectra analysis. The λmax was at 420 nm with an
absorbance of 1.233 and GC-mass spectra characterized 100 compounds. The
antimicrobial analysis gave positive inhibition of P. aeruginosa, S. aureus, C. albicans
and T. mentagrophytes with activity index of 0.2, 0.3, 0.4 and 0.2 respectively. The
cytotoxicity test showed that a concentration of 36.7185 grams in water of 350 ml of the
leaf extract was toxic based on the t-test of the live and dead cells by in-vitro analysis on
the lymphocytes from normal blood cells. (Hanif et al., 2019b)
Euphorbia hirta is an annual herb belongs to Euphorbiaceae family. It has been
found at warm and tropical regions mostly on roadsides and waste land throughout the
world and used as traditional medicines. Tawa-tawa contains lycosidal substance,
terpenoids, tannin, phorbic acid, fatty acids and sterols, Flavonoids including quercitrol,
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quercetin, and its derivatives. The extent or amount of these chemical constituents varies
in different parts of the tawa tawa plant and also depends on soil and climate condition.
Recently published reports showed that tea of tawa tawa plant is helpful as anti-malaria
and anti-dengue. Euphorbia hirta has been used for the remediation of respiratory disease,
some female disease and also other such as dysentery, jaundice, gonorrhoea, pimples,
tumors, digestive problems and children’s infections.
Plants are invaluable sources of pharmaceutical products and use of ethno
pharmacological knowledge enhances the probability of success in new drug finding
effort. Screening active compounds from plants lead to discover of new medicinal drugs
which have efficient protection and treatment roles against various diseases. Euphorbia
hirta Linn., (Euphorbiaceae) locally known as “tawa-tawa” and “maragatas”, a wild
herbaceous plant is very common in the Philippines. The plant has been widely
acknowledged for the treatment of cough, coryza, hay asthma, bronchial infections,
bowel complaints, worm infestations, and kidney stones in traditional medicine. In recent
publications, tawa-tawa extracts are used to treat dengue fever by rural folks in the
Philippines but no scientific studies have been made. Ringworm is an infection by a
fungus that most often affects the hair, nails and superficial layers of the skin. The most
commonly noted fungal types seen in cats and dogs are Microsporum canis,
Trichophyton metagrophytes and Microsporum gypseum. Ringworm fungi can be
transmitted to humans, therefore owners of infected animals should considered
quarantining the pet indoors until the infection is cured. Precautions should be taken
while treating animals in order to prevent human infection and environmental

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contamination. Tawa-tawa had been reported in other countries as antifungal agent. It
was demonstrated that the plant extract has a strong antifungal activity but limited study
has been conducted in the Philippines yet. Hence, this study was conducted to determine
the efficacy of Tawa-tawa (Euphorbia hirta.) crude extract against M. canis, in vitro . (Ul-
Islam & Banday, 2021)
CHAPTER 3
METHODOLOGY
3.1 Research Design
The researchers will be using the experimental design, specifically Complete
Randomized Design (CRD) in conducting the study. Different concentrations of Tawa-
tawa (Euphorbia hirta) will be extracted.
The researchers will use an experimental set-up and will carry it out to
determine the toxicity of Tawa-tawa (Euphorbia hirta) on red onion (Allium cepa) root
tip chromosomes. Three treatments will be prepared where each treatment consists of 3
replicates per trial.
T1R2 T1R3 T2R1

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T2R3 T3R1 T1R1
T1R2 T3R3 T2R2
3.2 Materials
Table1. Presents the different materials used in the study.
Materials Quantity
Tawa-tawa 300 g
Red Onion 27 pcs.
Blotting Paper 50 pcs.
Aceto Carmine 150 mL
Distilled Water 650 mL
3.3 Equipments/Apparatuses and Utensils
Table2. The different Equipment/Apparatuses and Utensils used in the study.
Equipment/Apparatus and Utensils Quantity
Beakers 9 pcs. (50mL)

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Cups 27pcs. (50mL)
Knife 1 pc.
Stirring rod 9 pcs
Container 2 pcs.
Meter Ruler 8 pcs.
Watch Glass 10 pcs.
Test Tubes 27 pcs.
3.4 Plant Identification and Collection
A total of 300g of Tawa-tawa herbs under the same condition will be collected
from Brgy. Poblacion, New Lucena between 6:00-8:00 A.M.. Only herbs without
adhering particles associated with damaged leaves and parts will be used. The herbs will
be washed using running water and drained to be prepared for exraction. The herbs will
be identified by Mr. Cristie S. Sumbilla, T-III, a Biology and Research Teacher, from the
New Lucena National Comprehensive High School.

18
Fig.2. Photograph of Tawa-tawa (Euphorbia hirta) herb
3.5 Plant Exraction
Three hundred grams of fresh whole Tawa-tawa plant (including roots) are
washed thoroughly, then carefully place these into the boiling pot. For every 100 g of
Tawa-tawa, boil for 15 minutes with 500 mL amount of water. Tawa-tawa will be
extracted through exraction. Let the concoction cool, then dispose leaves and stalks after
strain. Pour the liquid into a big container and cool. (Tayone, W. C. et al., 2014)
3.6 Planting and Preparation of Onions
A total of 27 red onions with the average size of medium and same condition will
be utilized in the study. The medium sized plant will be sun-dried for two weeks and the
dried roots present at the base of the onion bulbs will be carefully shaved off with a sharp
knife to expose the fresh meristematic tissues. These bulbs will be placed into a container
of fresh distilled water for the growth of red onions and to protect the primordial cells
from drying up. After a specific amount of time, the bulbs will be removed from the
container and placed on blotting papers to remove excess water. Three replicate bulbs
will be used for each test sample and control (distilled water) and the best two bulbs will
be chosen at the approximate time for examination. The base of each of the bulbs will be
19
suspended on the water sample in the 25 ml test tubes in the dark for 12 days. At the end
of the exposure period, the roots with the best growth will be removed with a forceps and
shall be utilized for chromosomal preparation. (Cabuga Jr et al., 2017)
3.7 Preparation for Treatment
The red onions will be distributed to three concentrations. These will be replicated
three times each concentration. The Tawa-tawa extract as the treatment will be prepared
by getting them ready with their corresponding treatment: 100%, 50%, and 0%. The first
can will be contained with 100% of Tawa-tawa extract as treatment one. In the second
can, 50% of Tawa-tawa extract will be placed as treatment two. The third can will be
contained with 0%, also known as the negative control, distilled water.
3.8 Measurement of the Root Length Mean
The root lengths of onion bulbs from each different treatment (100%, 50%, and
0%) will be measured after 72 hours of the test using a meter ruler. The root length mean
for each concentration will be calculated by adding the root lengths of individual onion
20
bulbs in that concentration and dividing by the number of onion bulbs. (Gessesew, W. S.,
2015)
3.9 Measurement of the Mitotic Index
Analyzing the mitotic index of Allium cepa (onion) typically involves counting
the number of cells undergoing mitosis relative to the total number of cells observed in a
particular sample. This will be measured after 5-10 days of exposure from the treatments.
First, obtain root tips from onion bulbs, as they contain actively dividing cells. Second,
stain the root tips to make the chromosomes visible under a microscope. Then, use a
microscope to observe the prepared slides. Count the number of cells in different stages
of mitosis (prophase, metaphase, anaphase, telophase) and the total number of cells. After
that, calculate the percentage of cells in mitosis by dividing the number of cells in mitosis
by the total number of cells counted, then multiplied by 100. And lastly, record the
observed data and analyze the mitotic index by comparing different samples, treatments,
or conditions of tawa-tawa (Euphorbia hirta) extract. Statistical analysis can help draw
conclusions about factors influencing mitotic activity. (Singh, 2016)
Data Analysis
The data on root length and chromosomal preparations will be presented as mean
values with standard error of the mean (SEM). Statistical analysis will be performed
21
using appropriate statistical software or tools such as T-test to assess the variability in the
data and determine the significance of the results. The values of root length mean and
mitotic index will be calculated by frequency count and mean. Differences between
corresponding controls and exposure treatment groups will be evaluated for statistical
significance, typically at a significance level of 5%. (Bonciu, E. et al., 2018)
Statistical Treatment
The statistical tools that will be used in this study are the frequency count, mean
and T-Test.
Frequency Count. The frequency count will be used for measuring of mitotic
index to determine the number of cells when exposed in different concentrations of
Tawa-tawa extract.
Mean. The mean of the root lengths will be computed to find out the average
value of roots’ growth when classified according to the different concentrations which
they were exposed.

22
T-test. T-test will be used to determine the significant difference among different
root length means and mitotic index when exposed in the different concentration of
Tawa-tawa extract.
REFERENCES
Bonciu, E., Firbas, P., Fontanetti, C. S., Jiang, W., Karaismailoğlu, M. C., Liu, D.,
Menicucci, F., Pesnya, D. S., Popescu, A., Romanovsky, A. V., Schiff, S.,
Ślusarczyk, J., De Souza, C. P., Srivastava, A., Şuţan, A. N., & Papini, A. (2018).
An evaluation for the standardization of theAllium cepatest as cytotoxicity and
genotoxicity assay. Caryologia, 71(3), 191–209.
Devi, P., Singh, P., & Kansal, S. K. (2020). Inorganic pollutants in water. Elsevier.
Essa, M. M., Akbar, M., & Guillemin, G. (2016). The benefits of natural products for
neurodegenerative diseases. Springer.

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Feretti, D., Zerbini, I., Zani, C., Ceretti, E., Moretti, M., & Monarca, S. (2014). Allium
cepachromosome aberration and micronucleus tests applied to study genotoxicity
of extracts from pesticide-treated vegetables and grapes. Food Additives and
Contaminants, 24(6), 561–572. https://doi.org/10.1080/02652030601113602
Gessesew, W. S. (2015). Growth Parameters of Onion (<i>Allium cepa</i>
L. var. Cepa) as Affected by Nitrogen Fertilizer Rates and Intra-row Spacing
Under Irrigation in Gode, South-Eastern Ethiopia. Agriculture, Forestry and
Fisheries, 4(6), 239.
Hanif, M. A., Nawaz, H., Khan, M. M., & Byrne, H. J. (2019). Medicinal plants of South
Asia: Novel Sources for Drug Discovery. Elsevier.
Hassan, G. M., & Yassein, A. a. M. (2014). CYTOGENOTOXICITY EVALUATION
OF WATER CONTAMINATED WITH SOME TEXTILE AZO DYES USING
RAPD MARKERS AND CHROMOSOMAL ABERRATIONS OF ONION
(Allium cepa) ROOT CELLS. Egyptian Journal of Genetics and Cytology, 43(1),
39–57.
Henry, G., & Janssen, W. (2018). Snap beans in the developing world. CIAT.
Khan, Z., Ansari, M. Y. K., & Shahwar, D. (2022). Induced genotoxicity and oxidative
stress in plants. Springer.
Larramendy, M., & Soloneski, S. (2015). Toxicity and hazard of agrochemicals. BoD –
Books on Demand.
Manoharachary, C., Tilak, K., Mallaiah, K., & Kunwar, I. (2016b). Mycology and
Microbiology (A textbook for UG and PG courses). Scientific Publishers.

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Peter, K. V. (2015). Handbook of Herbs and spices. Elsevier.
SAFA guidelines: Sustainability Assessment of Food and Agriculture Systems. (2014).
Food & Agriculture Organization of the UN (FAO).
Shigyo, M., Khar, A., & Abdelrahman, M. (2018). The Allium genomes. Springer.
Singh, R. J. (2016). Plant cytogenetics. CRC Press.
Tayone, W. C., Tayone, J. C., & Hashimoto, M. (2014). Isolation and Structure
Elucidation of Potential Anti-Dengue Metabolites from Tawa-tawa (Euphorbia
hirta Linn.).
Tardiff, R. G., & Rodricks, J. V. (2013). Toxic substances and human risk: Principles of
Data Interpretation. Springer Science & Business Media.
Wall, B. H. (2021). Charles Pettigrew, first bishop-elect of the North Carolina Episcopal
Church. Hassell Street Press.

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NEW LUCENA NATIONAL COMPREHENSIVE HIGH SCHOOL

1.1 The Problem and its Background

induce chromosomal aberrations in root meristems of Allium cepa.

preventing of the more prevalent and relevant diseases nowadays.

Furthermore, they have easily distinguishable genetic endpoints, such as

chromosomal aberrations (CAs), changes in ploidy and sister chromatid exchanges

(Sataphaty and Swammy, 2015).

Tawa-tawa herb or scientifically known as Euphorbia hirta linn plant, common

grasslands, roadsides and pathways. It is widely used in traditional herbal medicines

is rich in bioactive compounds like phenolic and flavonoids which may be

responsible for its anti-dengue properties.

(Euphorbia hirta linn) can affect in reducing the dengue properties.

1.2 Statement of the Problem

The main purpose of the study is to evaluate the toxicity of Tawa_tawa

(Euphorbia hirta linn) on Red onion (Allium cepa) root tip.

Tawa-Tawa (Euphorbia hirta) extract?

concentration of Tawa-Tawa (Euphorbia hirta) extract?

water), 100% and 50% concentration of Tawa-Tawa (Euphorbia hirta)

4. Is there a significant difference between the mitotic index of Red Onion

water), 100% and 50% concentration of Tawa-Tawa (Euphorbia hirta)

50% concentration of Tawa-Tawa (Euphorbia hirta) extract.

2. There is no significant difference between the mitotic index of R ed Onion

50% concentration of Tawa-Tawa (Euphorbia hirta) extract.

1.4 Conceptual Framework

Independent Variable Dependent Variable

Tawa-tawa (Euphorbia hirta):

50% Tawa-tawa herb extract + 50%

100% Tawa-tawa herb extract Mitotic Index

Negative control (distilled water)

1.5 Significance of the Study:

1. Community. This study provides further knowledge in agricultural practices or

ecological considerations in community, potentially influencing on how Tawa-tawa

(Euphorbia hirta) is managed or used in certain environments.

3. Medical Professionals. This study provides knowledge on what is the toxicity

affects the health of the humans.

provide additional literatures involving studies about toxicity of Tawa-tawa (Euphorbia

research about related matters involving toxicity of Tawa-tawa (Euphorbia hirta).

1.6 Scope and Delimitation

This study is limited on determining the toxicity of Tawa-tawa (Euphorbia hirta)

(Allium cepa) root tip.

included in the statistical analysis.

1.7 Definition of Terms

1. Tawa-tawa (Euphorbia hirta), is abundant in open grasslands and waste

responsible for its medicinal properties. (Wall, 2021)

(Essa et al., 2016)

2.1 Related Literature

2.1.1 Allium cepa

biochemical processes, leading to fatal results. Numerous potentially mutagenic

changes in the genetic material. Many thousands of toxic chemicals including

pharmaceuticals products, domestic and industrial wastes, pesticides and petroleum

exposure to chemicals. (Devi et al., 2020)

Genotoxicity is to determine the magnitude of gen etic risk to man by an

environmental agents/ chemical under a specified level of exposure. Unfortunately, the

considerations. (Detels et al., 2017)

Even the epidemiological approaches used to detect genotoxic and carcinogenic

chemicals. In order to identify the harmful effects of substances in different

environment. (Nations & Organization, 2019)

Higher plants constitute an important material for genetic tests to monitor

environmental pollutants. However this feature is due to the possibility of assessing

monitoring, especially in contaminated aquatic environments. (Khan et al., 2021)

2.1.2 Euphorbia hirta

E. hirta, commonly known as tawa-tawa, is a small annual herb common to