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Journal of Chemical and Pharmaceutical Research, 2015, 7(9):782-786
Research Article
ISSN : 0975-7384
CODEN(USA) : JCPRC5
782
Phytoremediation effect of Ricinus communis, Malva parviflora and
Triticum repens on crude oil contaminated soil
Sakina Saadawi
*1
, Marwa Algadi
1
, Amal Ammar
2
, Salah Mohamed
3
and Khairi Alennabi
4
1
Department of Natural Product, Faculty of Pharmacy, University of Tripoli, Libya
2
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Tripoli, Libya
3
Department of Microbiology, Faculty of Pharmacy, University of Tripoli, Libya
4
Biotechnology Research Center, Tripoli, Libya
_____________________________________________________________________________________________
ABSTRACT
Oil and its derivatives spills have been a major issue across decades and it is hard to biodegrade even though there
are many techniques are being developed to clean up petroleum contaminated soil. Phytoremediation has long been
applied as a treatment technology that is cost-effective, ecologically friendly and efficient for the decontamination of
hydrocarbon pollution. In this study, four crude oil contaminated soil samples were collected from oil extraction
fields in Libya. Three plants were chosen (Malva punilora, Ricinus communis and Triticum repens) on 0.5% and 1%
crude oil contaminated soil .The chosen plant species were implanted individually in the contaminated soil pots. Soil
sample (triplicate) was taken from each pot at zero time, after 15 days. After 30 days and after 45 days of
experiment. Hexane was added to the soil samples, mixed, filtered and the absorbance was measured using
spectrophotometer at 360 nm. The results were compared to the standard curve to calculate the crude oil
concentration and percentage of removal. As a result the highest percentage of removal of 0.5 % crude oil
contaminated soil was by Triticum repens (94%) after 30 days of experiment followed by Malva parviflora(88.5)
and Ricinus communis(77 %). While in 1% crude oil contaminated soil pots experiment, the highest percentage of
removal was by Malva parviflora (89%) after 30 days of experiment followed by Triticum repens (80%) and Ricinus
communis (76 %). Triticum repens showed good results suggesting more field application.
Key words: Phytoremediation, crude oil, Triticum repens, Malva parviflora, Ricinus communis.
_____________________________________________________________________________________________
INTRODUCTION
Phytoremediation is a group of technologies that use plants for remediating soils, sludge, sediments and water
contaminated with organic and inorganic contaminants. Plants have evolved a great diversity of genetic adaptations
to handle the accumulated pollutants that occur in the environment. Growing and, in some cases, harvesting plants
on a contaminated site as a remediation method is a passive technique that can be used to clean up sites with
shallow, low to moderate levels of contamination. Phytoremediation can be used to clean up metals, pesticides,
solvents, explosives, crude oil, polyaromatic hydrocarbons, and landfill leachates[1]. This emerging technology
promises effective, inexpensive, and less intrusive clean up and restoration of oil-contaminated environments [2].
Plants can decontaminate oil polluted sites by direct uptake of petroleum hydrocarbons into their tissues, direct
biochemical transformation of petroleum hydrocarbons and by mycorrhizal fungi and the activity of soil microbial
consortia [3].Plants that are resistant to crude oil toxicity such as black poplar and willows as well as miscanthus
grass (elephant grass) have been found to be effective in the remediation of oil polluted soil[4]. In the marsh
environment Spartina patens, Sagittaria lancifolia, Spartina alterniflora and Juncus roemeriannus are considered
ecologically and economically important in phytoremediation [5].
Sakina Saadawi et al J. Chem. Pharm. Res., 2015, 7(9):782-786
______________________________________________________________________________
783
Fossil fuels represent primary energy source in the global industry. This make a big threat on environmental
pollution, and serious ecological damage. Fuel by-products and spills in sites where storage, transport, distribution,
refining, consumption and the existing industries related fossil fuel can cause harm. About five million tons of crude
oil and refined oil go into the ecosystem every year due to anthropogenic sources from oil spills[6]. Contamination
of soil and water with hydrocarbons poses a major ecological and human health problem that needs an effective and
affordable technological intervention. Many sites stay contaminated with no treatment in sight because it is very
expensive to clean them with the available technologies [7].
With these aspects the present study was aimed to determine the percentage of crude oil removal by three plant
species (Ricinus communis, Malva parviflora and Triticum repens).
EXPERIMENTAL SECTION
Apparatus
UV-Visible spectrophotometer 6505 UV/VIS. (JENWAY), magnetic stirrer, beaker (50 and 100 mL), adjustable
pipettes (1-10, 10-100, and 100-1000 µL), sensitive balance, fume hood, autoclave and incubator were obtained
from Faculty of Pharmacy, University of Tripoli. Organic solvent toluene was obtained from Merck, Germany.
Plant collection
Three plant species were selected and collected during winter season. Selection was based on its availability,
resistance to the harsh environmental and soil conditions such as shortage of water and high temperature. These
species were Ricinus communis, which known locally as castor, Malva parvifloraor Malva and Triticum repens or
what’s known as Star).
Soil Preparation
According to the modified method of Ilyina et al. (2003)[8], soil was collected from Tripoli habitant area. In
separate pots, 3kg of soil was added. There were two main groups of concentrations (0.5% and 1%) as shown in
Chart (1), each contain four pots. The first potof each concentration of crude oil was used as a control (soil + crude
oil), the second, third and fourth werefor the effect of plant phytoremediation(Ricinus communis, Malva parviflora
and Triticum repens individually + crude oil). Crude oil (0.5% and 1%w/w) was added to the soil in thetwo groups
of pots (respectively) and mixed thoroughly. Total pots numbers were 4 for each group.
All pots were left under a shade, at temperature ranging from 10-20 °C, protected from rain and humidity
maintained everyday by spraying with distilled sterilized water to avoid dryness. Samples were analyzed at zero
time, after 15 days, 30 days and 45 days. The analysis was to determine the percentage of degradation of
hydrocarbon.
Chart 1: Experimental design for the effect of phytoremediation at 0.5% and 1% crude oil contaminated soil
Sakina Saadawi et al J. Chem. Pharm. Res., 2015, 7(9):782-786
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784
Measurement of Total Extractable Hydrocarbon Content
At zero time, 1 g (triplicates) of soil in control pots (contain 0.5%and 1% w/w of crude oil) was weighed and
transferred to dry, clean test tubes. Into this was added 10 mL of hexane, shook well and allowed to settle down for
30 min. The hexane-oil extracts were placed in cuvette wells and its absorbance was determined using
spectrophotometer at 360 nm wavelength for crude oil. The wave length was chosen after screening of several
dilutions of crude oil in the spectrophotometer. The best absorbance was at 360 nm. Then, all pots were sampled in
triplicate by the same way mentioned above at 15 days, 30 days and 45 days period.
Absorbance was converted to concentration by comparing it with standard curve. A calibration curve was obtained
by measuring absorbance of dilute standard solution of crude oil. Total hydrocarbon content was calculated after
reading the absorbance of extracts from the spectrophotometer, exploiting from calibration curve and multiplying by
an appropriate dilution factor (Osuji et al., 2006).
Calculation of percentage of removal of crude oil from soil
The percentage of degradation of crude oil was calculated by comparing the concentration results of the test with
those of the control using the formula:
Percentageofremovalofcrudeoilsamples = Concentrationofcontrol– Concentrationoftest
Concentrationofcontrol 100
RESULTS AND DISCUSSION
The percentage of crude oil removal from soil
A triplicates of soil in all pots in two groups (0.5% and 1% w/w of crude oil) was added 10 mL of hexane. The
absorbance of hexane-oil extracts was determined using spectrophotometer at 360 nm wavelength for crude oil. The
sampling was at zero time, 15 days, 30 days and 45 days period.
After conversion of absorbance to concentration by comparing it with standard curve, total hydrocarbon content was
calculated.
The result of percentage of removals of 0.5 % crude oil contaminated soil group of pots affected by the chosen plant
species(Ricinus communis, Malva parviflora and Triticum repens) were plot in figure 1. As the result showed, the
percentage of removal of 0.5 % crude oil concentration, Triticum repens or Star and Malva punilora shown to be the
most effective, reaching 94% of crude oil removal followed by Ricinus communis (77%). The crude oil percentage
of removal increased with time during the experiment for the three chosen plants.
.
Figure 1: Percentage of removal of 0.5 % crude oil contaminated soil by Ricinus communis, Malva parviflora and Triticum repens at 15
days, 30 days and 45 days of experiment
88.50%
94.00% 94%
85.70% 88.50%
94%
55%
77% 77%
0.00%
10.00%
20.00%
30.00%
40.00%
50.00%
60.00%
70.00%
80.00%
90.00%
100.00%
15 days 30 days 45 days
% of removal
Triticum repens Malva punilora Ricinus communis
Sakina Saadawi et al J. Chem. Pharm. Res., 2015, 7(9):782-786
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785
The result of percentage of removals of 1 % crude oil contaminated soil group of pots affected by the same chosen
plant species showed that same above pattern was obtained but with less percentages (Figure 2).
.
Figure 2: Percentage of removal of 1 % crude oil contaminated soil by Ricinus communis, Malva parviflora and Triticum repens at 15
days, 30 days and 45 days of experiment.
From the results, the three plant species Ricinus communis, Malva parviflora and Triticum repens showed to be
effective in removal of 0.5 % of crude oil contaminated soil. This can be explained that the group of techniques that
the plant use for phytoremediation process (phyto-accumulation, phytostabilization, rhizofiltration,
phytovolatilization, phytodegradation and rhizodegradation) were effective in crude oil removal. The time was
crucial in this process showing that the plant species were adapting with the new conditions in time of experiment
and the highest percentage of removal was obtained at the end of the experiment.
When crude oil concentration increased by double, the plant couldn’t adapt by the same level of crude oil removal at
the same time of experiment, which indicate that the plant species in this study need more time to degrade 1 % of
crude oil compared to 0.5 %. This may be taken in consideration in field study. As consistent with the previous
report by Basumatary et al [9] which was conducted to evaluate the efficacy of Cyperus rotundus (nut grass), that
could be effective in phytoremediation of crude oil contaminated soil. Total oil and grease (TOG) degradation and
microbial numbers were analyzed at different intervals i.e. 60,120 and 180 days in different percentages of crude oil
contaminated soil. In presence of crude oil, TOG content in soil, C. rotundus could decrease up to 50.01% in
vegetated pots during 180 days proving the efficacy of this plant species for use in phytoremediation.
This can be explained as the dissipation of petroleum contaminants in the rhizosphere is likely the result of enhanced
microbial degradation. Plant roots may encourage rhizosphere microbial activity through exudation of nutrients and
by providing channels for increased water flow and gas diffusion. Phytoremediation of crude oil in soil was
examined in other study using carefully selected plant species monitored over specific plant growth stages. Four
sorghum (Sorghum bicolor L.) genotypes. Soil contaminated with crude oil. All vegetated treatments were
associated with higher remediation efficiency, resulting in significantly lower total petroleum hydrocarbon
concentrations than non-vegetated controls [10].
Carpet grass (Axonopus compressus) was also examined in the phytoremediation management of oil impacted soil in
Ubeji and Alesa Eleme communities of Niger Delta region of Nigeria. The study employed an experimental research
design that involved the use of Axonopus compressus in the management of oil-impacted soil sites of Ubeji and
Alesa Eleme. The experiment lasted for four months. Axonopus sp. was used for the treatment of the crude oil
impacted sites. Laboratory analysis of the soil samples was conducted to determine the effect of phytoremediation
on hydrocarbon. The study revealed that the use of Axonopus sp. resulted in 66% loss of hydrocarbon from crude
oil-impacted soils of Ubeji and Alesa Eleme. The implication of the findings is that Axonopus sp. has the tenacity to
phytoremediate hydrocarbon concentration in soil effectively in any geographical region of the world [11].
46.00%
80.00%
94%
74.00%
89.00% 92%
21%
76%
84%
0.00%
10.00%
20.00%
30.00%
40.00%
50.00%
60.00%
70.00%
80.00%
90.00%
100.00%
15 days 30 days 45 days
% of removal
Triticum repens Malva punilora Ricinus communis
Sakina Saadawi et al J. Chem. Pharm. Res., 2015, 7(9):782-786
______________________________________________________________________________
786
Sang-Hawn et al.(2007) [12]concluded that hydrocarbon degrading bacterial populations increased rapidly during
the first four weeks of 14 weeks testing period. They proposed this finding that it may be considered as an indicator
for the feasibility of oil-contaminated soils degradation. However, with increasing of time, due to the oil-resistant
components with high chain and within less remaining nutrients, the bacteria growth and oil degradation
decreased[13]. Van Gestel et al. (2001)[14]reported a significant increment of the oil-polluted soil degradation in
bacteria population.
CONCLUSION
We conclude that, among the three selected plants, Triticum repens showed the highest crude oil degradability or
percentage of removal followed by Malva parvifloraand Ricinus communis during the period of experiment. As the
concentration of crude oil in soil increased the ability of plant phytoremediation decreased. All chosen plants
showed ability to degrade crude oil. These phytoremediation abilities were tested under the laboratory condition but
not at the field. Hence, field study of the above mentioned plants on higher crude oil concentration could be useful
in practical phytoremediation approaches and reduction of the risk from crude oil to human health.
Acknowledgement
The authors thank the Mr. Khairi Ali Alennabi, biotechnology research center (BTRS), Tripoli, Libya for his help
during the preparation of the experiment. The authors thank Mrs Marwa Ahmed Algadi, University of Tripoli and
Mr. Yahya Sharief Faculty of Pharmacy, University of Tripoli for provided lab facility.
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