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ProEnvironment
ProEnvironment 6 (2013) 582 - 586

Original Article

HPLC Assessment of Polycyclic Aromatic Hydrocarbons

Removal by Cluj Napocas Active Sludge Wastewater
Treatment Plant
ALHAFEZ Livia1, Nicoleta MUNTEAN2, Edward MUNTEAN*3, Dumitru RISTOIU1
1Babe-Bolyai

2University

University Cluj Napoca, Mihail Kogalniceanu St., no. 1, Cluj-Napoca, Romania

of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Mntur St., no. 3-5,
400372 Cluj - Napoca, Romania

Received 1 November 2013; received and revised form 15 November 2 013; accepted 26 November 2013
Available online 30 December 2013

Abstract
This paper presents the results obtained studying the PAHs removal in a conventional active sludge wastewater
treatment plant located in Cluj Napoca Romania, in which wastewater with a predominantly organic character is treated
using complex physical-chemical-biological processes. The collected samples were filtered to remove suspended matter,
and then PAHs were extracted using liquid-liquid extraction with hexane and determined by high performance liquid
chromatography using an Agilent 1100 system with both diode array and fluorescence detection. Separations were
accomplished using an Envirosep PP column and a mixture of acetonitrile: water as mobile phase. Detection limits ranged
from 0.05 g/kg for naphthalene to 0.001 g/kg for the other 13 of PAHs. The obtained results were expressed by the
cumulative amount of 14 compounds which ranged from 0.116 g/kg in the influent of the wastewater treatment plant to
0.0108 g/kg in the effluent. The amount of carcinogenic PAHs (benzo [a] anthracene, benzo [a] pyrene, benzo [b]
fluoranthene, benzo [k] fluoranthene, chrysene, dibenz [a, h] anthracene and indeno [1, 2, 3-c,d] pyrene) ranged from
0.0591 g/ kg in the influent to 0.002 g/ kg in the effluent. Overall PAHs removal ranged from 9.31% to 60.55 %.
Research showed that naphthalene, acenaphthene, fluorene, phenanthrene and anthracene are removed effectively during
wastewater treatment, whereas for benzo (b) fluoranthene, benzo (k) fluoranthene, benzo (a) pyrene, benzo (g, h, i)
perylene and dibenz (a, h) anthracene, indeno (1, 2, 3-cd) pyrene the recorded values were lower.
Keywords: polycyclic aromatic hydrocarbons, PAH, active sludge wastewater treatment plant, high performance
liquid chromatography.

1. Introduction
Polycyclic aromatic hydrocarbons (PAHs) are
a large group of organic compounds with two or
more fused aromatic rings, in linear, angular, or
cluster arrangements, which do not contain
heteroatoms [1, 20]. The polycyclic aromatic
hydrocarbons are of concern because of their
ubiquitous occurrence and high carcinogenic
potential [14].
* Corresponding author.
Tel.: +40 264 596384; Fax: +40 264 592 793
e-mail: edimuntean@yahoo.com

582

They represent benzene condensation

products
[22],
from
which
naphthalene,
acenaphthene, fluorene, phenanthrene, anthracene,
pyrene, benz (a) anthracene, chrysene, benzo (b)
fluoranthene, benzo (k) fluoranthene, benzo (a)
pyrene, benzo (g,h,i) perylene, dibenz (a,h)
anthracene, indeno (1,2,3-cd) pyrene are listed as
priority pollutants by the United States
Environmental Protection Agency and the European
Union, being also the target compounds in our
researches [2, 16].
The municipal wastewater treatment plant of
Cluj-Napoca is located on the left bank of the river

ALHAFEZ Livia et al./ProEnvironment 6(2013) 582 - 586

Somesul Mic downstream of Cluj-Napoca,

occupying 13 hectares, at 4647'34"N and
2340'57"E. The wastewater treatment plant consists
from a mechanical stage, a biological stage, a sludge
line and auxiliary installations, being designed for a
flow of 2170 L/s.
The technological process of urban
wastewater treatment includes the following phases:
mechanical treatment of urban wastewater, urban
wastewater biological treatment, and sludge
treatment. While an important effect of wastewater
treatment processes is to concentrate the organic
pollutants in the sewage sludge, the extent of the
removal depends on the properties of the organic
species. The municipal wastewater treatment plant
of Cluj-Napoca is located on the left bank of the
river Somesul Mic downstream of Cluj-Napoca,
occupying 13 hectares, at 4647'34"N and
2340'57"E.
The wastewater treatment plant consists from a
mechanical stage, a biological stage, a sludge line and
auxiliary installations, being designed for a flow of
2170 L/s. The technological process of urban
wastewater treatment includes the following phases:
mechanical treatment of urban wastewater, urban
wastewater biological treatment, and sludge
treatment. While an important effect of wastewater
treatment processes is to concentrate the organic
pollutants in the sewage sludge, the extent of the
removal depends on the properties of the organic
species. PAHs have a strong lipophylic character and
their water solubility decreases with the increase of
ring numbers [18]. PAHs with high molecular
weights are generally adsorbed to particles [22] while
low-molecular PAHs are relatively volatile [17].

PAHs can suffer photo-degradation and react

with other pollutants, such as sulfur dioxide,
nitrogen oxides, and ozone [3, 5, 8]. PAHs can react
with ozone, hydroxyl radicals, nitrogen and sulfur
oxides, and nitric and sulfuric acids, which affect
the environmental fate or conditions of PAHs [9].
Most PAHs are fluorescent [13]; the extended pielectron electronic structures of PAHs lead to
fluorescence spectra [12, 21]. PAHs removals
ranging from 33 to 100% from a specific treatment
plant were reported [19]; the removal efficiencies
for individual low molecular weight PAHs varied
from 84% to levels at which the compounds were
not detected in effluents [4, 15].
For the analytical determination, water
solubility is an important property. Most PAHs are
soluble in non-polar organic solvents and are barely
soluble in water [6]. PAHs water solubility
decreases with the increase of the number of fused
benzene rings and with angularity [7].
Various analytical techniques have been
developed to determine PAHs in soil [23], water,
food and air [11]. The methods include liquid column
chroma-tography, UV spectrophotometry, TLC,
spectro-fluorimetry, RP-HPLC with UV detection
and GC-MS.Since chromatographic separations are
primarily based on differences in physical properties,
the considerable variability described above for
PAHs makes sampling, sample preparation, and
analysis especially challenging [10].
The main objective of this study was to
evaluate PAHs removal efficiencies, based on liquidliquid extraction with hexane and high performance
liquid chromatograph with both diode array and
fluorescence detection.

Figure 1. A representative HPLC chromatogram of PAHs from the wastewater treatments plant influent

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ALHAFEZ Livia et al./ProEnvironment 6(2013) 582 - 586

2. Material and Method

PAHs in less than 30 minutes (fig. 1). Detection

limits ranged from 0.05 g/ kg for naphthalene to
0.001 g/ kg for the other 13 of PAHs. The
correlation coefficients for all analytes calibration
curves proved good linearity (correlation coefficients higher than 0.998).

Samples were collected from the influent and

the effluent of Someseni wastewater treatment plant.
The standard technique SR EN ISO 17993:2004 was
used; samplers were filled without air bubbles,
being stored at 4 50C until analysis.
Samples were filtered to remove suspended
matter, then PAHs were extracted using liquidliquid extraction with hexane and determined by
high performance liquid chromatography using an
Agilent 1100 system with both diode array and
fluorescence
detection.
Separations
were
accomplished using an Envirosep PP column (125
4.6 mm I.D) and a mixture of acetonitrile: water as
mobile phase, leading to baseline separation for 16

3. Results and discussions

The obtained data proves that PAHs removal
within the studied wastewater treatment plant occurs
(tab. 1), the most effective removal proving to be for
pyrene, chrysene, and benz (a) anthracene. In all
cases, influent PAHs concentrations were
significantly higher than effluent concentrations.

Table 1. PAHs recorded concentrations [g/ kg]

PAHs

Min.Conc.
- influent
0.0137
0.0049
UDL
0.0017
0.0002
0.002
0.0023
0.0001
0.0001
0.0011
0.0009
UDL
0.0006
0.0001

Naphtalene
Acenaphtene
Fluorene
Phenanthrene
Anthracene
Pyrene
Benz(a)anthracene
Chrysene
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(a)pyrene
Dibenzo(a,h)anthracene
Benzo(g,h,i)perylene
Indeno(1,2,3-c,d)pyrene
*UDL under the detection limit

Max.Conc.
- influent
0.0403
0.0124
0.0076
0.0137
0.0008
0.0423
0.0145
0.0292
0.0053
0.0013
0.0049
0.0022
0.0045
0.0018

As a general observed trend, the removal

efficiencies for high molecular weight PAHs were
higher than for low molecular weight PAHs (fig. 2).
C comp fin (ppb)-effluent

Table 2. Cumulative PAHs status [g/ kg]

0.0292

Sampling month
0.0145
0.0076
0

0.00170.0008

Max.Conc.
- effluent
0.0013
UDL
0.0127
0.0023
UDL
0.0057
0.0008
0.0097
0.0023
0.0009
0.0009
0.0003
0.0008
0.0022

Analyzing the cumulative PAHs status in

time, a quite strong variation was observed from
month to month for the period February May
(tab.2), the overall PAHs removal ranging from
9.31% to 60.55 %.

C comp fin (ppb)-influent

0.0423

Min.Conc.effluent
UDL
UDL
0.0082
0.0008
UDL
0.0007
0.0005
UDL
0.0006
0.0003
0.0003
0.0002
UDL
0.0002

0.0053

February
March
April
May

0.0049
0.0045
0.0022
0.0018

0.0012

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14PAHs
(influent)
0.0436
0.1160
0.0711
0.0311

14PAHs
(effluent)
0.0264
0.0108
0.0109
0.0133

4. Conclusions
The presence of PAHs in water bodies is a
major concern at both locally and globally level due
to the threat they pose to ecosystems.
The HPLC method used within our study
meets the required sensitivity and the selectivity,

Figure 2. Comparative situation of PAHs in influent

and effluent

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ALHAFEZ Livia et al./ProEnvironment 6(2013) 582 - 586

with high recoveries (between 91 - 99%) and good

reproducibility (CV between 3.22 - 4.99%).
Research
showed
that
naphthalene,
acenaphthene,
fluorene,
phenanthrene
and
anthracene are removed effectively during
wastewater treatment, whereas for benzo (b)
fluoranthene, benzo (k) fluoranthene, benzo (a)
pyrene, benzo (g,h,i) perylene and dibenz (a,h)
anthracene, indeno (1,2,3-cd) pyrene the recorded
values were higher, these being resistant to the
biological treatment.

aromatic hydrocarbons (PAHs) in oil shale processing

wastes: current practice and new trends. Oil Shale, 26(1),
59 - 72.

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