Remediation of Anthracene: A short comparative study on Biological and

Physicochemical approach
Abhrajyoti Tarafdar1, Alok Sinha2

1
Research Scholar,Department of Environmental Science and Engineering. Indian School of
Mines,Dhanbad
2
Assistant Professor,Department of Environmental Science and Engineering. Indian School of Mines,
Dhanbad

Abstract
Polycyclic aromatic hydrocarbons (PAHs) are added to the environment by the pyrolysis of petroleum at high
temperatures as well as in acetylene synthesis from natural gas, refinery operations, etc. Anthracene is a PAH
with a high pollution potential and health risk. It has been used as model substrate in studies on the
environmental degradation of PAHs, since the structure of anthracene is found in carcinogenic PAHs. This study
is aimed to compare the degradation of anthracene by advance oxidation process and by bacterial species
isolated from PAH contaminated colliery sites, near Jharia, Jharkhand. Analysis of anthracene degradation is
estimated by Fluorescence Spectrophotometry near 400 nm (emission). The percentage removals of anthracene in
both cases are measured. The biological process gave 82.8% removal in 6 days, whereas, the physicochemical
oxidation process gave 84% degradation in 30 mins.

Keywords: PAH Degradation, Anthracene Degradation, Bacterial remediation of Hydrocarbons, AOP of PAH

1.Introduction

The industrial revolution after the World War II has extremely changed the life norms. Modern lifestyles are
largely depending on various industries where specifically, chemical & petroleum industries are extensively
contributed in the various life branches. However, when the relation between modern lifestyles civilization and
the environment is to be evaluated, disappointingly disproportional growth is to be appeared accompanied with
great tension on the different environmental components. Anthropogenic contaminants caused by human
activities throughout the worldwide industries coupled with other natural contamination feed such tension and
are seriously threatening the environment. We must find the cure of this disease for our Mother Earth.

Polycyclic aromatic hydrocarbons (PAH) are formed by various kind of incomplete combustion of organic
material in natural and anthropogenic processes (like volcano activity, pyrolysis). Hence they are distributed
ubiquitously. In various studies has been detected presence of PAHs in air, water, and soil. In some cases there
were found the concentration of these contaminants more 10 g/kg (Lundstedt, 2003). Concerning their
mutagenic and carcinogenic potential, the United States Environmental Protection Agency (EPA) listed 16 PAH
as priority pollutants.

PAHs in contaminated soils, waters, and sediments present a significant risk to the environment and human
health, as they are hydrophobic compounds, and they persist mainly in ecosystems through their low water
solubility (Bonnet et al., 2005).

For treatment of PAHs there are mainly three kinds of technologies:

1. Physical treatment: extraction with subcritical and supercritical fluids, surfactants, vegetable oil, solvents, etc.
2. Chemical treatment: oxidative treatment where ozone and/or hydrogen peroxide (with or without iron ions)
are employed (Gmez-Alvarez et.al., 2012).
3. Biological treatment: potential aromatic ring degrading microbes are used for bioremediation.

Anthracene is one of those hundreds of PAH and was chosen for study because of its relative toxicity (Jacques
et.al., 2005). It is a solid polycyclic aromatic hydrocarbon consisting of three fused benzene rings. There are
very less informations available from studies on humans to tell what effects can result from being exposed to
individual PAHs at certain levels. However, breathing PAHs and skin contact seem to be associated with cancer
in humans. So, the need of remediation of anthracene is pertinent.
The present study deals with the comparison between the biological degradation and degradation by advance
oxidation process (ozonation).

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 1. Analytical Procedure
2.1 Biodegradation studies
Soil samples for isolating microorganisms of interest were collected from Jharia, Jharkhand. Samples were
collected from three different waste drainage sites outside of coal washeries. Three different vials are used for
collecting the samples.

For isolating the bacteria of interest the basic principle followed is to grow the cultures in a media which has
anthracene as the sole carbon source. So that, the bacterial species have to use anthracene in order to survive.
Anthracene (250mg/litre) is dissolved in Acetone & let the Acetone evaporate from the conical flask. Next, the
Bushnell Haas Broth made in the same flask leaving Anthracene as sole carbon source. This process will help
Anthracene to suspend in water base media, resulting comparatively high accessibility for microorganisms.
Serial diluted (10-5) sample soil is added in the media and kept for 8 days in 120 rpm 35oC. The whole process
is repeated several times for fine tuning.

After a 16 days growth in selective media, the colonies are plated in R-2A Broth (0.312gm in 100 ml distilled
water with 2% agar) and incubated for 24h at 32oC. Successfully isolated species of bacteria are shown hereby
(Fig. 1).

Fig. 1. The isolated Microbial Strain.

Fig. 2. The flowchart of the analytical procedure of the degradation kinetics of anthracene

The biodegradation of anthracene can be measured by a degradation kinetics given above (Fig. 2).

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2.2 Ozonation Process

The study of degradation of anthracene by ozonation is held with anthracene in the same media condition of
aqueous solution. Anthracene concentration in the media given is 10mg/liter. The percentage of ozone in the
passing air was 30%. Different runtime was occurred for ozonation of anthracene in the media.

2.3 Anthracene Analysis

It was shown anthracene has fluorescence property, 250nm (excitation) and 406nm (emission) (Krivobok et al.,
1998).

In this report anthracene analysis was determined by Fluorescence Spectrophotometer (Model: F-2500 FL
Spectrophotometer, Serial Number: 1866-005, ROM Version: 4000 01).

2. Results and Discussions

3.1 Biodegradation

The lag phase of the isolated species was found till 0-17th hours (Fig. 3). Thereafter the growth was seen in the
exponential phase for 25-120th hours. Then it enters the stationary phase. Bacterial growth reaches at its
maximum at around 120th hours. Simultaneously anthracene degradation is also shown hereby.

0.5 0.0035
Biomass
0.45
0.003
0.4
0.35 0.0025

Residual anthracene conc. (gm/l)

0.3 0.002
Biomass (gm/l)

0.25
0.2 0.0015

0.15 0.001
0.1
0.0005
0.05
0 0
0 50 100 150 200
Time (h)

Fig. 3 Anthracene Concentration and Biomass Dry Weight Vs. Time plot.

The residual anthracene has been used by the bacterial biomass. From the graph it is clearly evident that max
anthracene degradation took place in first 24 hours. Then its gradually decreasing.As the biomass is increasing
the residual anthracene concentration is decreasing, which supports the ability of the isolated strain to degrade
anthracene effectively.

= 82.8%

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3.2 Ozonation

The graph for different runtime of ozone through the media containing anthracene is given hereby (Fig. 4),

600

500
Fluorescence Emmison Intensity

400

300

200

100

0
0 5 10 15 20 25 30 35

Time (min)

Fig. 4. The Fluorescence emisson of anthracene

From the above graph we can interprete that first five minutes of ozonation degraded most of the anthracene and
after twenty minutes of ozonation we can see that the degradation came to a saturation end point as the thirty
minutes of ozonation does not produce any effectively more degradation.

So, here the percentage removal of anthracene by ozonation is,

3. Conclusions
Though, percentage of degradation is slightly greater in the advance oxidation process but, it should be
mentioned that biological method is almost free of cost where ozonation method is much expensive. Also in
practical fields, where the oil refineries produce PAH, the biological method of degradation can be more
acceptable because there time is not a factor but money is. While PAH with less than five rings are usually
biodegradable under aerobic conditions (Yuan et al., 2000), the use of highly condensed PAH by bacteria as sole
carbon and energy source has not been reported properly yet (Kornmuller & Wiesmann, 2003). Therefore, a
pretreatment by ozonation may be added to the process for a partial oxidation of highly condensed PAH, which
is followed by a biological treatment of the formed oxidation products and the accompanying oil for a higher
rate of PAH removal.

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