TECHNOLOGICAL INSTITUTE OF THE PHILIPPINES

COLLEGE OF ENGINEERING AND ARCHITECTURE

CHEMICAL ENGINEERING DEPARTMENT
363 P.CASAL ST., QUIAPO, MANILA

EVALUATION OF CYTOTOXICITY AND GENOTOXICITY

OF CELLULOSE ACETATE FROM CIGARETTES FILTERS
USING ALLIUM CEPA TEST IN HYDROPONICS SYSTEM

Submitted by:

BALINO, Russel Christian M. – CHE31S2

CAPELLAN, Seth Christian D. – CHE31S2
GARCIA, Raffy V. – CHE31S1
PURGANAN, Jerilly B. – CHE31S1

Submitted to:
Engr. Lina Dela Cruz

Date submitted:
SEPTEMBER 3, 2020
 A. INTRODUCTION

A1. BACKGROUND OF THE STUDY (Mr. Balino, Russel Christian)

Cigarette filters (CFs) are the most common litter on earth, with an estimated 4.5 trillion discarded
annually. Apart from being unsightly litter, chemicals can emit leach from consumed cigarette filters and
can lead to severe human health and environment issues. Unsurprisingly, several studies have reported
that these cigarette filters which is made up of cellulose acetate are the most common item retrieved by
clean-up activities in public areas such as beaches and parks.

Cigarette filters are chemically composed of 95% cellulose acetate (CA) taking the form of a
monofilament tow in of the material along with abundant known chemicals and additives which are
hazardous waste when consumed. The cigarette filters are about 30% of the whole cigarette’s length and
usually contains traces of tobacco (Hemamalini et al., 2019). A filter is purposely designed to reduce the
quantity of fine particles, smoke, and tar that can be inhaled when combusted and to reduce the severity of
the smoke and keep the tobacco particles away from the smoker’s mouth.

According to Poppendieck (2020), a used butt or filter, if someone touched a used cigarette filter, it can give
off the equivalent of up to 14% of the nicotine that an actively burning cigarette emits. Harmful compounds
were also identified as eight of the hundreds of chemicals typically emitted from cigarettes, including four
that are on the FDA list of harmful and potentially harmful constituents. These harmful substances can
cause asthma, hay fever and certain reactions such as rashes, vomiting, headache, tight chest, hives and
worsening of eczema.

Among the polymers derived from renewable sources, the cellulose acetate is highly desirable as a raw
material in the production of biodegradable superabsorbent polymers because it is the most abundant
natural polymer on earth. Furthermore, Senna et. al. (n.d.) suggests that cellulose acetate has the
capability of being used as an alternative soil in our modern agriculture, the hydroponics system. Plants in
soil have to rely on many factors such as the physical climate availability of water, presence of pests and
disease organisms, weeds, and soil structure and chemistry for their growth and crop yield. Hence,
hydroponics is widely used to produce many vegetables (tomato, lettuce, cucumber and pepper) and many
other ornamental crops. Soilless cultivation is a method of growing plants using mineral nutrient solutions,
in water, without soil, is supported by using inter medium such as perlite, rockwool, clay pellets, peat moss,
or vermiculite instead of the root system (Fan et al., 2012).

Meric et al. (2011) also stated that soilless cultivation is widely used in order to improve the control of the
growing environment and avoid uncertainties in the water and nutrient status of the soil. This system also
aids in saving irrigation water and fertilizers, thereby appreciably increasing the water use efficiency by the
crop. This study aims to utilize the cigarette filter wastes and involve it into the hydroponic system as an
alternative inert medium in growing healthy vegetable plants using allium cepa bioassay.

On this matter, bioassays with plants have been considered highly sensitive and simple for the monitoring
of chemical compound cytotoxic effects. Allium cepa (onion) has been indicated as an effective test
organism for evaluating toxicity at the cellular level (Caritá & Marin-Morales, 2008). Due to its kinetic
properties of proliferation and for having large chromosomes and few in number (2n = 16), Allium cepa test
has been made convenient. It facilitates their analysis in the detection of micronuclei and mitotic spindle
abnormalities as well as the verification of changes in the cell division rate (mitotic index). The mutagenic
activity of cigarette filters used and rejected by the factories will be subjected to Allium cepa bioassay in a
hydroponic system. This study aims to evaluate the genotoxic and cytotoxic effects of the filters to Allium
cepa L. Root meristem cells.

A2. SIGNIFICANCE OF THE STUDY (Mr. Capellan, Seth Christian)

This research study creates an innovative system of a new alternative inert medium derived from
cigarette filter wastes. Research studies like this develop the value of resourcefulness and open-
mindedness of the researchers and also homeowners who have less space area to plant but is interested
in growing plants. If proven effective, this will serve as a guide in production of simple and safe
hydroponics system at home. Furthermore, this study will help lessen the wastes produced annually and
will evaluate the genotoxicity and cytotoxicity of the filters.

A3. SCOPE AND DELIMITATION OF THE STUDY (Mr. Capellan, Seth Christian)

This research study entitled “Evaluation of Cytotoxicity and Genotoxicity of Cellulose Acetate from
Cigarette filters using Allium cepa Test in Hydroponics System” is limited only to the cigarette filter’s ability
to grow good quality vegetables including the toxicity effects depending on the composition of the filter in
the study . Also, the potential of the plants to be edible is not examined in this study. Other related matter
such as the fertilizers and are no longer honored in this study.

B. THEORETICAL FRAMEWORK

B1. REVIEW OF RELATED LITERATURES (Mr. Garcia, Raffy & Ms. Purganan, Jerilly)

B1.1 TOBACCO SUPPLY CHAIN

From tobacco cultivation, curing, processing, cigarette manufacturing and distribution, to use and
final disposal, the tobacco industry’s supply chain is global and extensive, but fragmented across regions,
and at every stage a range of resources and materials are required (World Health Organization, 2018).

Figure 1. Global cigarette consumption and consumption supply chain

Source: World Health Organization, 2018

Tobacco cultivation takes place across over 125 countries. It involves such activities like seedling
production, management of soil, irrigation, utilization of agrochemicals, harvesting, and burning of residues
of crops. Upon harvest, tobacco leaves undergo curing in order to reduce moisture content and ensure
duration and capacity of preservation. Majority of the produced tobacco for cigarette production are
subjected to flue-curing.
The process of flue-curing involves the drying of the green tobacco within closed buildings along with
furnaces driving the heat into the barn. The most common sources of energy for this type of process are
wood or coal and these are the main inputs for the initial stage for the production of tobacco. After curing, it
follows grading and blending of the tobacco leaf, the removal of non-tobacco constitutes, stemming of raw
materials, additional drying stages, packing, and temporary storage. It ensures ultimately the total control of
moisture content deterioration. In addition, water is also one of the main inputs being utilized at the
processing and storage at the facilities.

In the span of two to three years, the processed tobacco will be shipped to the manufacturing companies
where it is reorganized into cigarettes. The manufacturing process of cigarette involves the input of
secondary materials such as cigarette filters, cigarette papers, tipping papers, packaging materials,
flavorings, and adhesives; and requires large amount of considerable and plausible amount of energies.
Distribution of manufactured and process tobacco includes the customers within the domestic and
international companies, with the use of proper and appropriate transportation, directly to warehouses and
outlets.

In addition to huge amounts of manufactured trillions of cigarettes sticks, the outputs of the chain supply
may include tons of waste and leachate which considers the hazardous waste and the post-consumer
wastes which thereof include cigarette filters which contributes on one of the top tons of greenhouse gas
emissions (World Health Organization, 2018).

B1.2 COMPOSITION OF CIGARETTES

Marinello, et al. (2019) described the cigarette as a small cylinder of finely cut tobacco leaves rolled
in a thin paper. Commercially made cigarettes in modern generations are composed of the following
elements shown in Figure 1.
 Tobacco blend dried and cut into a millimeter strip;
 Rolling the paper to cover the cylindrical shape of tobacco, and adhesives;
 A filter composed mostly of cellulose acetate; and
 Tipping paper, usually an ornamental paper, to cover the filter which is the cellulose
acetate part.

Figure 2. Components of Cigarette

Source: Marinello et al., 2019

B1.3 CHARACTERIZATION OF CIGARETTE FILTERS

 Cigarette filters (CF) are chemically composed of 95% cellulose acetate (CA) taking the form of a
monofilament tow in of the material along with abundant known chemicals and additives which are
hazardous waste when consumed. The cigarette filters are about 30% of the whole cigarette’s length and
usually contains traces of tobacco (Hemamalini et al., 2019). A filter is purposely designed to reduce the
quantity of fine particles, smoke, and tar that can be inhaled when combusted and to reduce the severity of
the smoke and keep the tobacco particles away from the smoker’s mouth.

The filter of cigarettes is composed of poorly degradable fine-sized white fibers amassed together with
approximately 12,000 per filter and with approximately 30,000 dernier which are distinguished by a cross-
section of Y-shape wherein it is not perpendicular to the flow within the angle ranging from 18° to 49° (Du
et al., 2015). These fibers are synthetic plastic which look like cotton (Marinello et al., 2019).

Cellulose acetate is treated with the plasticizer during the manufacture of cigarettes that generally improves
the proportionality of its mechanical properties to its mechanical performance. Plasticizers are known to in
terms of acetic (glycerol triacetate), trimethylene glycol diacetate (TEGDA), and its mixtures. Filters are also
contaminated with adhesives, flavors, and fragrances in addition to plasticizers, to enhance its market
performance. On the other hand, adhesives are composed of polyvinyl acetate, ethylene vinyl acetate,
cellulose acetate, and mixtures thereof. In addition, the flavors and fragrances are additives that can be
absorbed by the filter material in different constituents in the form of nicotine and menthol (Schiraldi, 1996).

Cellulose acetate is not easily biodegradable although it is a photodegradable kind of polymer. The basic
degradation or decomposition of cellulose acetate is an initial deacetylation process by means of chemical
hydrolysis and acetyl esterases, enzymes that are commonly found in microorganisms. The backbone of
the cellulose is degraded then with cellulose. The intertwined network of fibers with the plasticizers made
the process of disintegration and decomposition very slow and difficult to overcome. Nowadays,
sustainable disposal methodology for this waste are not yet known (Benavente et al., 2019).

B1.4 TOXICITY OF CIGARETTE FILTERS

A filter is purposely designed to reduce the quantity of fine particles, smoke, and tar that can be
inhaled when combusted and to reduce the severity of the smoke and keep the tobacco particles away
from the smoker’s mouth. Also, its purpose is to reduce the harshness of the smoke and to keep the flakes
of tobacco out of the smoker’s or inhaler’s mouth. The filter captures and retains these toxic substances
(Marinello et al., 2019). Thus, the toxicity of CFs stems from the thousands of carcinogenic agents trapped
after usage. This can leach into the environment and eventually enter into the food web (Abu-Danso et al.,
2019).

B1.5 STATISTICS REGARDING ENVIRONMENTAL IMPACT OF CIGARETTES

Cigarette filters are considered one of the most constituents that litters un urban areas with an
estimation of 4.5 trillion of which 22 to 46% of it are visible litters. On the other hand, it is found that the
environmental impact of the CG waste is directly related to its chemical composition with an approximately
130 chemicals have been reported to be present in CBs. However, many more chemicals (40,000–
100,000) have been identified in cigarette smoke, some of which could potentially be retained in CBs.

This waste is made of nearly 95% microscopic-sized fibrous CA. The remaining 5% constitutes of the toxic
substances which include heavy metals such as iron, manganese, strontium, aluminum, lead, copper,
chromium, cadmium, and etc. Plasticizers are applied to bond the fibers. One cigarette filter soaked in a
liter of water for 96 hours is the Lethal Concentration 50 for test fish exposed to cigarette filters’ leachates
(Benavente et al., 2019).

According to the World Health Organization (2018), the tobacco industry produces six trillion cigarettes that
are consumed by one billion smokers worldwide annually. In addition to the six trillion cigarette sticks, the
outputs of the supply chain included 25 million tons of solid waste (including hazardous waste and the post-
consumer waste such as CBs) on top of the millions of tons of greenhouse gas emissions.

The non-tobacco elements of cigarettes such as filters, cigarette paper, and packaging all carry a burden
on the environment, especially when one considers the vast quantities used in the manufacturing of 6
trillion cigarettes annually. More than 1 Mt of filters and about 2.15 Mt of packaging are estimated to be
used by the tobacco industry in a year (this excludes cardboard boxes for shipping). Furthermore, all the
post-consumer waste that these items become and which has to be treated or, most importantly, ends up
contaminating the environment, considerably aggravates tobacco’s environmental footprint. Table 2.1 lists
the annual environment impact of global tobacco use and disposal.

Table 1. Total annual environment impact of global tobacco uses and disposal

Impact Category Unit Amount in Millions

Climate change kg CO2 eq 870
Terrestrial acidification kg SO2 eq 2.9
Freshwater eutrophication kg P eq 0.3
Marine eutrophication kg N eq 1.0
Human toxicity kg 1,4-DB eq 534
Terrestrial ecotoxicity kg 1,4-DB eq 6
Freshwater ecotoxicity kg 1,4-DB eq 29
Marine ecotoxicity kg 1,4-DB eq 24

Source: World Health Organization, 2018

B1.6 POTENTIAL RECYCLING SOLUTIONS FOR CIGARETTE FILTERS

According to the literature review conducted by Marinello et al. (2019), the following are potential
recycling solutions for used CFs:

 Polarz et al. (2002) designed the modification of cigarette filters into micrometer and nanometer
porous carbon structures. CB’s derived from smoked or unsmoked cigarettes were reorganized
into non-graphitic carbon structures. The resulting property of material displayed a well-defined
mesoporosity, with unique characteristics from the fiber of CA used as a progenitor and with good
potential application in the field of gas purification.

 Escobar & Sanz (2017) analyzed the performance of smoked and unused CFs as absorbing
materials. Each sample was obtained by collecting 10 filters inside an impedance tube. The
samples differed in the type of filter used and their physical characteristics (length and diameter).
All samples exhibit a significant and promising sound absorption in which it can be compared to
 some of the materials like foam and glass wool, which makes it a great potential for commercial
purposes.

 Teixeira et al. (2017) used CFs and waste from filter production in the cigarette industry for
cellulose pulp production. With the help of their treatment technology, they obtained a pulp from the
cellulose and a viscous dark liquor. The pulp can be utilized in the paper industry while the dark
liquor must undergo treatment because of its highly resistant organic content.

 Mohajerani et al. (2017) studied the possibility of incorporating cigarette filters (CFs) into asphalt
concrete for pavement construction, encapsulating CBs with different classes of bitumen and
paraffin wax. The chemical translocation from the CB’s to the environment is prevented by the
Encapsulation. Comparing the results obtained in the two cases with the control asphalt concrete
sample (no CBs), using mechanical and volumetric properties, including stability, flow, resilient
modulus, bulk density, maximum density, air voids, and voids in mineral aggregates as variables,
the proposed solutions were found to satisfy the requirements for light, medium and heavy traffic
conditions for bitumen samplers, while paraffin wax only satisfied the light traffic conditions for road
pavements. The proponents of this study identified the solution as a relevant and useful
contribution in decreasing the urban heat effect in the urban environments due to the reduction of
bulk density which is the increase in porosity and the reduction of thermal conductivity.

 Xiong et al. (2018) studied the use of CBs as a precursor for the development of supercapacitors,
which are one of the best potential energy storage systems thanks to higher energy and power
densities than conventional capacitors. The development of new application for the energy storage
of CBs produced carbon with a large specific capacitance and remarkable retention of capacitance
thorough the combination of CB’s and polypyrrole (PPy). PPy is a conducting polymer used as an
electrode material for supercapacitors. The remarkable potential storage performance obtained by
the proponents is due to the effect of synergy of the porous backbone structure which are derived
from CBs and the pseudo-capacitance behavior of PPy-derived nitrogen functionality.

B1.7 CELLULOSE ACETATE

Cellulose acetates (CAs) are the key esters of cellulose, that are obtained by the reaction of acetic
anhydride and acetic acid to cellulose in the presence of sulfuric acid solution. The most renowned form of
CA fiber contains an acetate group with approximately 2.00 - 2.25 of evert 3 hydroxyls. The cellulose
diacetate formed is known as the secondary acetate or simply as acetate. Cas can be used for different
applications depending on the process that will be applied (Fischer et al., 2008).

CAs are one of the first polymeric membrane material utilized for aqueous base separation specifically. CA
membranes exhibits remarkable toughness, high hydrophilicity, good desalination property, high flux
potential, and relatively low-cost material. Thus, CAs are being applied widely into different unit operations
and separation processes such as reverse osmosis, gas separation, microfiltration, and ultrafiltration. The
aforementioned separation techniques and technologies have been used in the domains of water
treatments, wastewater treatment, pharmacy and medicinal industry, and food processing (Bai et al., 2012).
B1.8 EXTRACTION OF CELLULOSE ACETATE FROM DISCARDED CIGARETTE FILTERS

In a study conducted by Benavente et al. (2019), three aqueous solutions, 5% NaCl (w/v), 5%
NaAc (w/v) and 0.02% H2SO4 (w/v), were used to wash three samples of ten discarded cigarette filters
(DCFs). Each sample was extracted with its corresponding solution.

Figure 3. Concentration of metals in aqueous solutions after extraction of DCFs

Source: Benavente et al., 2019

After that, each sample was extracted using absolute ethanol, diethyl ether, and hexane, successively.
Then, the cleaned butts were suspended in acetone and distilled water was added until a white precipitate
of cellulose acetate was formed.

Table 2. Organic compounds extracted from DCFs by absolute ethanol

Ethanol Extract
Compound
after NaCl after NaAc after H2SO4
(p-hydroxyphenyl)-phosphonic acid P P

m-cresol (F) P P

3-ethylphenol (A) P P

indole (F) P P

1,2,3-propanetriol diacetate P P

nicotine (A) P P

3-methylindole P P

2,4-bis(1,1-dimethylethyl)-phenol 1-pentadecanol (F) P P

1-tridecanol P

p-tertbutylphenol P
 phthalic anhydride P

methyl dodecanoate (F) P

methyl octanoate (F) P

1-tetradecene P

n-tridecanol P

Legend: P - present in extract, F - flavoring agent, A - adhesive

Source: Benavente et al., 2019
Table 3. Organic compounds extracted from DCFs by diethyl ether

Diethyl Ether Extract

Compound
after NaCl after NaAc after H2SO4
bis(2-ethylhexyl)phthalate (Pa) P P
eicosane (W) P P
hexadecane (W) P
butylhydroxytoluene (A) P
1,2,3-propanetriol diacetate P

Legend: P - present in extract, Pa - plasticizer, W - wax, A – antioxidant

Source: Benavente et al., 2019

Based on the results of the study, the most efficient methodology should include initial extraction steps with
aqueous solutions, starting with an H 2SO4 solution, followed by a NaCl solution. After that, an extraction
process using an organic solvent should be considered to eliminate the organic components of DCFs.
Finally, a precipitation step using the acetone-water system would purify further the CA.

B1.9 RECYCLING CIGARETTE FILTERS INTO CELLULOSE PULP

In the University of Brasilia, researchers recycle cigarette filters into cellulose pulp since cigarette
filters are mainly composed of cellulose acetate, a polymer. These cigarette filters undergo alkaline pulping
which is a lignocellulosic pulping process to improve the color of the resultant pulp. It is obtained by using
alkali-based substance on the cigarette filters. After conducting the process, the pulp strength increases by
conducting the delignification process between the temperatures 100-160 degree Celsius. The resulting
product of the process can now be used for textile and cellophane production.
Figure 4. Flowsheet of Hydrothermal Pre-treatment and alkaline pulping process
 Source: Jameel, H. (n.d)

B1.10 CIGARETTE FILTERS USED FOR GROWING ORNAMENTAL PLANTS

One of the solutions proposed by researchers from the Centre of Agro-Environmental Research “Enrico
Avanzi’ at the University of Pisa, was that cigarette filters containing cellulose acetate can be used to grow
ornamental plants using hydroponic cultivation. Consumed cigarette filters had undergone several
processes such as separation of the materials and chemical treatment for it to be suitable for growing
plants. After obtaining the product that has the highest tendency to support growth, several plants will
undergo germination on the products to find the best plant that can adapt to such system.

Figure 5. Ornamental plant growing with the use of cigarette filters

Source: Floral Daily, 2020

B1.11 Allium cepa L. ASSAY IN ASSESSING CYTOTOXICITY AND GENOTOXICITY

Over the last decades, environmental pollution with toxic chemicals has become a world-wide
issue due to rapid urbanization and industrialization. Heavy metal contamination is persistent in the
environment, which greatly affects the food chains, and it also leads to different health related problems
due to their toxicity. Hence, serious exposure threat is a real threat especially for living organisms. With
this, researchers develop ways and method, one of which is the use of onion plant ( Allium cepa L.), to
monitor the genotoxicity of different chemical contaminants and to measure the degree alterations in the
environment caused by mutagens.
In the study conducted by Amon and Firbas (2014), they showed that the Allium cepa L. plant is an
appropriate indicator to determine the potential genotoxic agents from the different samples taken from the
environment. The genotoxic level of the agents was measured by the structural changes of chromosomes
taken from the meristem cells of the young growing onion roots – for it is very sensitive to genetic damage
caused by chemicals, thus, it shows that it is a suitable model system for determining the potential
environmental cytogenetic of pollutants.
Alongside, A. cepa L. assays is commonly utilized bioindicator in genotoxicity testing because is it cost-
effective. It can provide valuable information about the presence of genotoxic and /or mutagenic
compounds present in the surfaces of different ecosystems. It assesses cytotoxic and genotoxic endpoints
such as nuclear alterations, chromosomal aberrations, mitotic index alterations, and root growth inhibition
(Wijeyaratne & Wadasinghe, 2019).
B1.12 EXISTING STUDIES THAT ASSESS THE CYTOTOXICITY AND GENOTOXICITY USING A. cepa L.
 Mangalampalli, Dumala, & Grover (2017) measure the toxicological efficiency of MgO
nanoparticles and macroparticles using A. cepa L. as an indicator. It was evaluated under various
bioassays on A. cepa root tip cells which includes oxidative stress, comet assay, and their uptake
or internalization profile. Results showed an increase in chromosomal aberrations and a decrease
in mitotic index compared to control cells. Comet analysis revealed a range of 6.8-30.1 over 12.5
-100 μg/mL DNA damage in terms of percent tail DNA, and it was found that to be more significant
at the exposed concentrations. A substantial growth in hydrogen peroxide generation and
superoxide radicals was observed in accord with the lipid peroxidation profile compared with
control. Thus, it revealed the exposure of MgO NPs showed greater toxicity on A. cepa L. than
MPs.

 Saben, et. al (2020) performed an A. cepa L. test for tomato (Lycopersicum esculentum) and chili
(Capsicum annum) as A. cepa L. is a helpful tool to test the genetic variations in plants. The extract
of these plants was used to grow the roots of A. cepa L. Examinations under microscope revealed
various chromosomal abnormalities at different level of treatment such as laggards, vagrant
chromosomes, stickiness, nuclear lesions, binucleated cells, c-mitosis, and giant cells. It was
concluded that the rate of cell aberrations increases as the metal stress also increases.

 Olorunfemi, et. al (2011) used A. cepa L. to investigate the potential genotoxic effects of the
effluents from the processing of cassava tubers into three popular Nigerian meals – garri, lafun,
akpu. They found out that there was a rapid decrease in mitotic index as the concentration of
effluents increases. The findings indicate that the contained substances in the cassava effluents
may cause harm may be toxic to the living organisms. Alongside, it may also possess threats to the
environment.
The subsequent is an existing research study that also adapt this method to assess discarded cigarette
filters’ toxicity:
  Montalvao, et.al (2018) used the meristem cells of A. cepa L. to assess and determine the
chromosomal and nuclear abnormalities of water containing smoked cigarette filters (SCF).
According to their data, plants exposed to SCB showed a large number of abnormal cells, high
inhibition index, low relative grow index, and high abnormality frequency at metaphase or
anaphase stage. It was discovered that the SBC leachate possess aneugenic and clastogenic
effect on the tested plants’ genetic material, either when they acted additively, synergistically, and
individually. Thus, it was concluded in their study that the disposal of cigarette filters in the
environment my cause cytotoxic, mutagenic, and genotoxic effects on plants.
These are some of the existing researches that utilize A.cepa L. to assess the cytotoxicity and genotoxicity
of certain substances. It only shows that this method is suggestively essential especially with the prevailing
condition of the environment. In this research, this method will be used to evaluate the cytotoxicity and
genotoxicity possessed by cigarette filters in hydroponics system.

B2. CONCEPTUAL FRAMEWORK (Mr. Balino, Russel Christian)

In this study, the researchers will be utilizing cellulose acetate of that cigarette filters derived from
rejected manufactured cigarettes and consumed cigarettes. The CFs will undergo the washing, extraction
using aqueous solutions, decantation, and sterilization processes. In addition, CFs will be used as an
alternative soil matrix in the hydroponics system. On this matter, bioassays with plants have been
considered highly sensitive and simple for the monitoring of chemical compound cytotoxic effects. Allium
cepa (onion) has been indicated as an effective test organism for evaluating toxicity at the cellular level
(Caritá & Marin-Morales, 2008). This study aims to evaluate the genotoxic and cytotoxic effects of the
cigarette filters to Allium cepa L. Root meristem cells.

INDEPENDENT VARIABLES

Amount of consumed cigarette

filters and factory reject filters
containing cellulose acetate

DEPENDENT VARIABLES

Number of chromosomal aberration

mitotic index
 INTERVENING VARIABLES

Temperature
Humidity
Amount of organic fertilizer

Figure 6. Conceptual framework of the study showing the

identified variables which will be present in the study

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