Phytoextraction of Mercury-Contaminated Soil using Cogon Grass (Imperata cylindrica)

Davao City National High School

F. Torres St., Davao City

An Official Entry for the Regional Science and Technology Fair

(Physical Science-Individual Category)

Researcher:
Avila, Edd Rashid Matthew

Consultants:
Orcheliza L. Paramo. Ph,D
Richard Jann B. Escobido, Chemist

Sherwin Fortugaliza, Ph. D

Research Adviser

November 2018
 Phytoextraction of Mercury-Contaminated Soil using Cogon Grass (Imperata cylindrica)

Edd Rashid Matthew Avila

Davao City National High School

F. Torres St., Davao City
______________________________________________________________________________

ABSTRACT

The global problem concerning contamination of environment as a consequence of human

activities is increasing. Most of the environmental contaminants are chemical by-products and
heavy metals such as Mercury (Hg). Mercury released into the environment makes its way into the
air, water, and soil. Mercury contributes to a variety of health effects such as decline in mental,
cognitive, and physical health of the individual. This study was conducted to determine the
phytoextraction potential of Cogon (Imperata cylindrica) Grass on mercury contaminated soil.
Ten plants in each pot ranging from 245 to 250 grams were placed and were observed for six (6)
weeks. Results showed using Atomic Absorption Spectroscopy (AAS) the plant samples has an
average Hg accumulation capacity of 40.763 ppm or 76.53 percent. It was also confirmed that the
said plant do survive in the said environment with a mean of 83.3% survival rate. Dependent
sample t-test confirmed that there is a significant difference in the amount of mercury before and
after the phytoextraction (t=28.421; p=.001). The amount of mercury decreased significantly with
a mean difference of 40.76 ppm from the pre-concentration of 53.263 ppm. Confirmatory test on
the root analysis showed that mercury was detected on the said part. This confirm that Cogon
Grass plant with its high Mercury accumulation capacity is a positive environmental footprint
making it an ecologically friendly sorbent for the development of phytoextraction technology.

Keywords: Atomic Absorption Spectroscopy, Imperata cylindrica, Mercury, Phytoextraction

______________________________________________________________________________

1. BACKGROUND OF THE STUDY pollutants for the surface water and land soil
because of their toxicity and being non-
Almost all the industrial and
biodegradable (Agarwal et. al, 2010).
commercial activities generates effluent
Mercury is a heavy metal, the second most
containing one or more heavy metal. Some of
toxic element in the world. The toxic effects
these in trace quantities serve as
of mercury depend on its chemical state and
micronutrient for the life processes of living
the route of exposure. Mining industries
organisms. But most of these beyond their
which are unregulated pose potential hazards
permissible limits are the most dangerous
to the environment due to the generated waste

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coming from the mining processes which
could create problems in air and water
pollution. Mining is one of the most
destructive man-made activities that brought
negative implications to the environment.
The tailing pond of the mining industries is
considered as a home of pollution and
contaminants that contributed negative
effects to the health of the environment. A
tailing pond may possibly contaminate water
bodies when runoff occurs during heavy rain
or through leaching into nearby rivers and
ground waters (Mazyck et. al, 2009). It has
been known for many centuries that mercury
is poisonous to animals and human beings,
but by Minamata mishaps (1953) of Japan,
extreme toxicity of Hg to humans became
evident (Sharma, 2011).
Utilization and exploitation of Hg and
Hg based technology coupled with
inadequate disposal of Hg-laden waste posed
an increasing threat on environment
contamination (Sindhu, 2014). Some of the
major industries discharging Hg in their
effluents are chlorine manufacturing,
switchgears/batteries, paints, fertilizers and
pharmaceuticals. Addition of Hg through
waste discharge in to natural bodies of water
is of serious concern today because of
probable entry of Hg in to the food chain
3
apart from possible direct ingestion (Shafeeq
et. al, 2012).
The researcher used Cogon grass
(Imperata cylindrica) to remove mercury in
contaminated soil. Imperata cylindrica
(Cogon Grass), it is a perennial rhizomatous
grass native to
Southeast Asia, India, Micronesia, Australia,
and Eastern and Southern Africa. It grows
from 0.6 to 3 m (2 to 10 feet) tall.
The leaves are about 2 cm wide near the base
of the plant and narrow to a sharp point at the
top; the margins are finely toothed and are
embedded with sharp silica crystals. The
main vein is a lighter color than the rest of the
leaf and tends to be nearer to one side of the
leaf. The upper surface is hairy near the base
of the plant while the underside is usually
hairless. Roots are up to 1.2 meters deep, but
0.4 m is typical in sandy soil. In other
countries, they consider this plant as a pest
(Bhakta et. al, 2011).
Many remediation techniques have been
employed to address the rising number of
heavy metal contaminated soils (Cholpecka
et al. 1996). Most of the traditional methods
such as incineration, vitrification,
electrokinetics and land filing are extremely
expensive. A promising and relatively new
technology for remediation of heavy metal
contaminated sites is phytoextraction. The
use of the plants (grasses in this study) in
phytoremediation may be cost effective and
could be environmentally friendly, but the
practical phytoremediation tool for the
control and remediation of Hg contamination Figure 1. Collection Site of Cogon
in soil using tropical grasses is not well Grass Plants
understood. The researcher used Atomic
Absorption Spectroscopy (AAS) for the
determination of heavy metal absorbed by the
samples (Lasat, 2012).

2. METHODOLOGY

2.1 Collection of Materials

The researcher obtained the soil
sample from the mining area located at New Figure 2. Collection of Cogon Grass
Plants
Leyte, Maco, Compostella Valley. The soil
sample was directly given by the mining
Figure 3. Collection of Mercury
workers as it was suggested and was assisted Contaminated Soil
by the DENR staff. Restriction in the mining
site was explained.

For maximum safety, Personal

Protective Equipment (PPE) was ensured.
The researcher collected Cogon grass
seedlings at Molave, Phase 3, Indangan,
Buhangin, Davao City, Philippines.

2.2 Preparation of Materials

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 The materials that were used in this
study are: 3 flower pots, Cogon grass
seedlings, contaminated soil, weighing scale,
and surgical gloves that will be borrowed at
Davao City National High School Science
Laboratory.
Figure 4. Weighing of collected
Mercury Contaminated Soil for each
trial
2.3 Plant Identification
The plant samples were authenticated
by Ms. Orcheliza L. Paramo, Ph.D of Davao
City National High School, located at F.
Torres St., Davao City.
2.4 Phytoextraction of Mercury
Cogon grass (Imperata cylindrica);
were grown in individual flower pots
containing 4 kg of mercury contaminated
soils with a concentration of exactly 53.263
ppm respectively. Young seedlings of Cogon
5
grass served as test plants for the said plants’
trials. Ten (10) plants were planted in each
pot with weight ranging from 245 to 250
grams in triplicate.
The phytoextraction potential of the
plants were carried out at Molave, Phase 3,
Indangan, Buhangin, Davao City to simulate
the environmental condition. Individual pots
were inspected daily for possible leakage that
may compromise the results of the study
(Visvanathan, 2010).
Figure 5. Measuring of each Cogon
grass plant’s height for each trial
(before)
All treatments were given exactly 150
mL of distilled water every four days for a
period of five weeks to keep the moisture
content of the soils at field capacity. Distilled
water was used in the experiment to make
sure that water is free from impurities,
chemicals and pollutants, which are potential
sources of contamination (Sindhu, 2014).
 $𝐴0 −𝐴𝑓 (
𝐴" = 𝐴0 x 100

Where 𝐴) is the amount of mercury

present in soil after the accumulation test, 𝐴*
is the amount of mercury present before the
accumulation test, +𝐴* − 𝐴) , is the quantity
where the net mercury accumulated, and 𝐴"
is the capacity rate of the Cogon grass as a
Figure 6. Planting of Cogon grass plant
bioaccumulator (Bhakta et. al, 2011).
in each pot of 3 trials (before)

Survival rate of the Cogon grass

(Imperata cylindrica) was also determined
using the formula (Henry, 2000):

./
𝑆" = . x 100
0
Where 𝑁* is the number of plants
before in each trial, 𝑁) is the number of is the
Figure 7. Cogon grass plants in each pot
of 3 trials (after) number of plants which survived in each

Mercury concentrations in the soil quadrant, and 𝑆" is the survival rate.

and plant samples were measured by atomic

absorption spectroscope. Analysis of Hg 2.5 Oven-dry Method

were done in plants and soil before planting, After the accumulation of the said

which will be served as the initial Hg plant, the researcher oven-dried the plant’s

concentration, and after harvesting, which part: roots, for 48 hours in 65-70⁰C to obtain

will be served as the final Hg concentration the constant weight (Henry, 2000). The

of the samples. To get the amount of mercury method was conducted at University of

compound that will be accumulated by the Immaculate Concepcion located at

plant will be calculated using the formula: Bankerohan, Davao City for the

6
determination of mercury concentration of
the said part.

Figure 10. Pre-concentration analysis

on the collected mercury-contaminated
soil

Figure 8. Cogon grass plant’s roots

Figure 11. Atomic Absorption

Spectroscope
2.7 Microscopy
Figure 9. Cogon grass plant’s root The Cogon grass roots and leaves
depth or length
were examined under 20X, 100X, and 2000X
2.6 Spectroscopy magnification using Olympus CX23 for the
The researcher directly brought the examination of its features and characteristics
soil sample for pretesting at Davao (Abdel-Wahab et. al, 2012).
Analytical Laboratory and for final testing as
the University of Immaculate Concepcion
located at Bankerohan, Davao City. And to
determine the mercury accumulated in soil,
before and after tests, they were analysed by
Atomic Absorption Spectroscope
(Kacalkova et. al, 2009). Figure 12. Cogon grass plant’s leaf on a
20X magnification

7

Figure 13. Cogon grass plant’s leaf on a
100X magnification
Figure 15. Cogon grass plant’s roots on
a 100X magnification
Figure 16. Cogon grass plant’s roots on
a 2000X magnification
2.8 Proper Waste Disposal
After the experimentation, all the
instruments that were borrowed from Davao
City National High School were returned
safely and the other variables used in the
8
experimentations were thrown safely in a
black plastic bag. The Mercury-contaminated
soil that were analysed on Davao Analytical
Laboratory and University of Immaculate
Concepcion Laboratory, were managed and
disposed properly by the said managements.
The researcher followed the RCRA standards
to create an added cost and represents a
potential drawback to the technology
(Moreno, 2004).
Figure 17. Disposal
2.9 Data Analysis
The researcher used T-test for Dependent
Sample Statistics to test if there is a
significant difference in the amount of
mercury accumulated in Parametric Test. The
researcher also used Mean Statistics to
calculate the average accumulated mercury
concentration capacity in each trial of the said
variable (Shafeeq et. al, 2012).
3 RESULTS AND DISCUSSION
 Table 1 presents the Mercury 2 10 7 70
Accumulation capacity of the Cogon grass 3 10 9 90

plant. The initial concentration of mercury of Mean 10 8.33 83.3

the soil was 53.263 ppm. After the

phytoextraction, the soil was sample has an Table 2 shows the survival rate of the

average mercury content of 12.5 ppm. Cogon grass plant in a Mercury-

contaminated environment in the span of 5

The least mercury concentration (ppm) weeks. It shows that 83.3% of the number of

accumulated by the said plant is 38.263 ppm plant’s samples in each trial or quadrant can

(71.84%) while, the maximum amount of survive in the said environment.

concentration accumulated by the said plant

is 43.263 ppm (81.23%). Dependent sample t-test confirmed
that there is a significant difference in the

Table 1. Mercury Sorption Capacity of amount of mercury before and after the
Cogon Grass phytoextraction (t=28.421; p=.001). The
Mercury
Pre- Post- amount of mercury decreased significantly
% Sorption
Trials Conc Conc
Absorbed Capacity
(ppm) (ppm) with a mean difference of 40.76 ppm.
(ppm)
1 53.263 10.0 81.23 43.263
2 53.263 15.0 71.84 38.263
Table 3. Test of difference in the Hg content
3 53.263 12.5 76.53 40.763 Mean t df P
Mean 53.263 12.5 76.53 40.763
Before 53.263 28.421 2 .001
After 12.50
The minimum mercury sorption
capacity is 38.263 and the maximum is
Confirmatory analysis was also
43.263 and an average of 40.763 ppm.
conducted using root analysis:

Table 4. Mercury Concentration Capacity of

Table 2. Survival Rate of Cogon grass plant
the Plant’s Part
Trials Pre- Post- Survival Hg-Conc.
Trials Part
treatment treatment Rate (%) (ppm)
1 10 9 90 1 Roots 0.93

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 Table 4 shows that Cogon grass
plant’s part: roots, do have concentrations of
mercury that has been accumulated. The
result presents that the mercury concentration
detected on the roots was 0.93 ppm.
Based from the results in the
experimentation, Cogon grass (Imperata
cylindrica) has a potential as a phytoextractor
and phytoaccumulator in removing mercury
compounds present in the soil specifically, in
the mining areas.
It is important to note that of the total
amount of ions associated with the roots, only
a part is absorbed into cells. A significant ion
fraction is physically absorbed at the
extracellular negatively charged sites of the
root cell walls (Henry, 2000). Thus, it is
possible that a plant exhibiting significant
metal accumulation into the root, to express a
limited capacity for phytoextraction. Binding
to the cell wall is not the only plant
mechanism responsible for metal
immobilization into roots and subsequent
inhibition of ion translocation to the roots.
This proves that the Cogon grass
(Imperata cylindrica) plant do accumulate
Hg. This is because that the mercury
accumulated by the Cogon grass plant
10
stacked up in the root part. Because of the
heavy metal’s charge, metal ions cannot
move freely across the cellular membranes,
which are lipophilic structures. Therefore,
ion transport into cells must be mediated by
membrane proteins with transport functions,
generically known as transporters.
Transmembrane transporters possess an
extracellular binding domain to which the
ions and is responsible for transporter
specificity. The transmembrane structure
facilitates the transfer of bound ions from
extracellular space through the hydrophobic
environment of the membrane into the cell
(Wang, 2004).
There are two mechanisms which are
responsible in the heavy metal transport from
the contaminated-soil to plant’s roots: 1.)
Convection or Mass Flow, 2.) Diffusion, and
3.) Plant’s root’s surface area (Lasat, 2012).
Due to convection, soluble metal ions move
from soil solids to root surface. From the
rhizosphere, water is absorbed by roots to
replace water transpired by the leaves. Water
uptake from rhizosphere creates a hydraulic
gradient directed from the bulk soil to the root
surface. Some ions are absorbed by roots
faster than the rate of supply via mass flow
(Mazyck et. al, 2009). Thus, a depleted zone
is created in soil immediately adjacent to the
root.
This generates a concentration
gradient directed from the soil solution and
soil particles holding the absorbed elements,
to the solution in contact with the root
surface. This concentration gradient drives
the diffusion of ions toward the depleted
layer surrounding the roots (Shafeeq, 2012).
And lastly, due to the high surface area of the
Cogon grass plant’s roots, the heavy metal
uptake is enhanced making it a very good
phytoaccumulator of a heavy
specifically mercury, the second most toxic
element on earth.
4 CONCLUSION
The present study indicated that Cogon
grass possessed many beneficial
characteristics to remove Hg
contaminated soil. It has a tolerant result and
could grow in soil contaminated with a high
Hg concentration from the mining area
(53.263 ppm). Cogon grass plant does have a
potential phytoextractor and phytoremediator
because of its high mercury accumulation
capacity with an average of 40.763 ppm. The
results show that the average
concentration of the contaminated soil is 12.5
ppm making the soil stable from the mercury
contamination. It has also proved that the
Cogon grass plants do survive in Mercury-
contaminated environment, specifically the
mining areas. The standard mercury
concentration of a healthy soil ranges from 0
to 16 ppm of the said metal. This is another
reason, why Cogon grass plant do have a
potential as a natural phytoaccumulator
(Sharma, 2011).
The important implication of these
findings is that Cogon grass can be used for
phytoextraction on sites contaminated with
metal, high levels of heavy metals, particularly on
Hg. It can be used to remove Hg, thus
gradually reducing the contamination levels.
It is difficult for anyone to avoid exposure to
the presence of harmful heavy metal in the
environment, particularly Hg, since most Hg
concentrations found in the environment are
from the results of human activities (Sindhu,
2014). Through the use the said grass to
phytoremediate Hg contaminated soil, human
exposure to Hg will be lessened since Hg
contributes to a variety of adverse health
effects, especially to children. The use of
Cogon grass is a very cost effective
environmentally friendly and practical
post- phytoremediation tool for the control and
remediation of Hg contamination in soil,
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since it is an invasive type of plant and an
abundant specie in the local and global areas.

5 RECOMMENDATIONS
Based from the results of the study, the
following recommendations are strongly
recommended:
1. Use Cogon Grass plant in
phytoremediation of Mercury-
contaminated soil or sites.
2. The Cogon grass’ leaves must also be
examined to determine the Mercury
concentration present on the said
plant’s part.
3. The same study should be conducted
but in a natural Mercury-
contaminated environment.
4. Same study can also be carried out but
should increase the number of trials
for a more liable finding.

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