PAPER www.rsc.org/jem | Journal of Environmental Monitoring

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Tenax-TA extraction as predictor for free available content of polycyclicaromatic hydrocarbons (PAHs) in composted sewage sludges†

Patryk Oleszczuk*

Received 11th January 2008, Accepted 2nd June 2008

First published as an Advance Article on the web 16th June 2008

DOI: 10.1039/b800532j

The usability of the solid phase extraction method with Tenax-TA adsorbent for the forecasting of

polycyclic aromatic hydrocarbon losses during sewage sludges composting has been evaluated in the

present study. Four municipal sewage sludges were composted for 76 days. The PAH content in the

sludges ranged from 3674 to 11 236 mg kg�1. After composting, a significant reduction of PAH content

was observed. The range of the PAH sum loss ranged from 29.3 to 48.6%. The share of fraction

extracted by Tenax-TA in the sludges at the onset of the composting ranged from 28.1 to 54.0% in

relation to the PAH sum, and from 0.31 to 100% for individual PAHs. After 76 days of composting, the

presence of Tenax-TA extracted PAHs was still observed in the sludges. A significant reduction of this

fraction was noted in the case of three sewage sludges. PAH’s Tenax-TA fraction in the case of one

sewage sludge did not undergo any changes during composting. A satisfactory prediction of PAH losses

by the Tenax-TA method was rarely observed. The best reflection of the PAH losses was observed in the

case of fluoranthene, anthracene and indeno[1,2,3-cd]pyrene.

Introduction

Speciation analysis plays an important role in analytical chem-

istry, ecotoxicology and biotechnology. It makes the recognition

of chemical and biochemical processes taking place in the natural

environment possible, and enables evaluation of the actual risks

for human and the whole environment. From the point of view of

ecotoxicology and bioremediation, it is important to determine

the fraction which is directly available for organisms to be

practically used for bioremediation progress forecasting as well

as risk assessment. It allows adjustment to the remediation

conditions to be made to achieve the most effective cleansing

process. In recent years, procedures have been sought which

allow for the precise determination of those pollutant fractions

that are available for microorganisms or invertebrates. This issue

is more broadly discussed in a number of review papers.1,2

Learning about the processes that influence bioavailability is at

the moment one of the key issues in environmental science in

terms of organic pollutant fate.2 Determination of the total

pollutants content is not always the same as the determination of

the real risk posed by their presence in the natural environment.

Sewage sludge composting is an alternative method for the

optimization of their properties, with the aim of using them

further for fertilization purposes.

Studies show that as a result of this above mentioned process,

a significant improvement in sewage sludge properties as well as

a reduction in the content of many organic pollutants can be

achieved.3–6 Polycyclic aromatic hydrocarbons (PAHs) are

common contaminants present in sewage sludges.7–10 Frequently,

Institute of Soil Science and Environmental Management, University ofAgriculture, ul. Leszczy�nskiego 7, 20-069, Lublin, Poland. E-mail:patryk.oleszczuk@ar.lublin.pl; Fax: +48 (81) 532 26 32; Tel: +48 (81)534 35 48

† Electronic supplementary information (ESI) available: Additionaltables (S1 and S2) and figure (S1). See DOI: 10.1039/b800532j

This journal is ª The Royal Society of Chemistry 2008

the exceeding of the allowable thresholds of contaminant content

in sludges is observed, and hence these sludges are excluded from

use in agriculture.11 The application of sewage sludges containing

contaminants into soils can result in their accumulation in the

soil12 as well as their uptake by plants.13 Taking into account the

mutagenic/carcinogenic and toxic PAHs properties, it is clear

that it is necessary to reduce contaminant content, for example

by composting, before they can be introduced into the soil.

Hence, it is of utmost importance to know: (1) the strength of the

interaction between contaminant and sewage sludge or compost,

and (2) the contribution of bioavailable forms occurring in

sludges and composts. The above will allow us to determine the

actual environmental risk, as well as any potential possibilities of

contaminant content reduction in sewage sludges. Even though

the influence of the composting process on the changes in the

PAH content is a popular subject nowadays, there is little

information in the literature on the PAH bioavailability in

sewage sludges and composts.14,15 The method using the Tenax-

TA adsorbent makes forecasting possible of both biodegradation

progress16,17 and bioaccumulation and ecotoxicity18–20 of organic

pollutants. Hence, besides the evaluation of the bioavailable

contaminant fraction, it also gives an actual picture of their

potential ecotoxicological influence on living organisms.

The aim of the present study was to (1) evaluate, using

a Tenax-TA adsorbent, the influence of the composting process

on the content of bioaccessible21–23 forms and (2) examine the

possibility of application of this method for the evaluation of

progress in PAH losses in the sewage sludges composted.

Materials and methods

Composting process

Four sewage sludges collected from municipal sewage treatment

plants were chosen for the present study. Each sludge was

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composted in two parallel bins (30 L) for 76 days. Ventilation

was provided through air distribution tubes. In order to increase

oxygen inflow, the composted material was additionally mixed

once a fortnight. Subsamples for analysis (200 g) were collected

directly before the onset of the experiment (sewage sludge) and

after composting (compost). Full characteristics of the sewage

treatment plants and the composting process (changes of

temperature, mass losses and pH) were presented in other

papers.6,24

Samples for analysis were collected with a stainless steel corer

throughout the whole bin height. All samples were air-dried and

milled to be representative.

PAHs analysis

Samples (15 g) were extracted by dichloromethane (2 � 40 mL) in

an ultrasonic bath (Sonic-3, Polsonic, Poland). The extracts were

centrifuged, decanted and evaporated to dryness. The residues

were then dissolved in 4 mL of acetonitrile : water mixture (1 : 1

v/v) and purified by solid phase extraction (SPE) using C18

octadecyl columns (JT Baker-Mallinckrodt, Germany) as

described elsewhere.24 Ten PAHs from the EPA list were

analysed: phenanthrene, anthracene, fluoranthene, pyrene,

benzo[a]anthracene, chryzene, benzo[b]fluoranthene, benzo[a]

pyrene, benzo[ghi]perylene and indeno[1,2,3-cd]pyrene. A quali-

tative and quantitative analysis of PAHs was carried out on the

liquid chromatograph with UV detection (TermoSeparation

Products). The reversed-phase, high performance liquid chro-

matography (HPLC) system consisted of a Spectra Series P100

pump (Thermo Separation Products) coupled with a Spectra

Series UV100 detector (Thermo Separation Products) and

a computer PC. For the separation of 16 PAHs, an analytical

Spherisorb S5 PAH (250 � 4.6 mm I.D., 5 mg by Schambeck SFD

GmbH, Germany) column with chemically bound C18 phase,

was used. The column was installed in a thermostatted oven at

30 �C (LCO 101, ECOM, Czech Republic). Acetonitrile and

water (82 : 18, v/v) were used as eluent solvents at a flow rate of

1 mL min�1 (in isocratic conditions). Detection was carried out at

254 nm. Elution of all PAH was carried out by 60 min. Data

acquisition and analysis was performed using the Clarity Lite

Chromatographic Station (DataApex, Czech Republic).25

Recoveries for the total procedures (sample preparation,

extraction and SPE) ranged between 81 to 90% for individual

PAHs. Precision expressed as relative standard deviation (RSD)

was below 12%. The concentrations reported here have therefore

not been corrected for losses. A method blank did not show

reagent or equipment contamination with PAHs.

Tenax-TA extraction

The method using Tenax-TA was originally proposed by

Cornelissen et al.23 In the study presented, the method as

described by Cornelissen et al.18 and Hulscher et al.20 after

modifications was used. Desorption was determined at 20 �C by

means of a single Tenax-TA solid phase extraction method.

During desorption, a mixture of Tenax-TA (3 g), sewage sludge

or compost (2 g dry wt), and deionised water (70 mL) was

constantly shaken in a 100 mL separation funnel for 6 h. After

desorption, the Tenax beads and the sewage sludge or compost

884 | J. Environ. Monit., 2008, 10, 883–888

samples were separated. The separation of Tenax from the

mixture was easily achieved because it floated on the top of the

water. Sewage sludge sank to the bottom of the separation

funnel, where it was removed together with the water, while the

beads of Tenax-TA remained on the walls of the separation

funnel. The collected Tenax beads were extracted with 2 � 30 mL

of acetone for a few seconds. The extracts were evaporated to

dryness and then residues were dissolved in 1 mL of acetonitrile :

water mixture. PAH content was determined in accordance with

the method described above. The efficiency of the Tenax-TA

extraction to remove the PAHs from water was determined in

a separate experiment by extracting spiked water samples, and

recoveries were 98–110%. The water was spiked with ten ana-

lysed PAHs. PAHs recovery was determined at three different

concentration levels depending on the contaminant tested.

Quality control

The procedural blank was determined by going through the same

extraction and cleanup procedures for each series of samples.

None of the analytical blanks were found to have detectable

contamination of the monitoring PAHs, and thus the results

were not blank corrected. The mass balance in the case of Tenax-

TA extraction was determined by comparing the total amount of

chemical desorbed by Tenax-TA and the amount remaining in

the sewage sludge (as determined in the DCM) after Tenax-TA

extraction with the initial measured amount in the sample. The

mass balance ranged from 91 to 102% for all sewage sludges.

Data analysis

All reported concentrations of PAHs in samples are on a dry-

weight basis of sewage sludge (determined by drying the samples

for 24 h at 105 �C) and are the average of triplicate extraction.

The relationships between the phycico-chemical properties of

sewage sludges/compost and PAHs were determined by corre-

lation analysis with Statistica 5.0. Significance was set at *P <

0.05. Statistically significant differences between the results were

evaluated on the basis of standard deviation determinations and

by analysis of variance (ANOVA).

Results and discussion

Total PAHs content and changes during composting

The content of the PAH sum analysed in individual sludges

ranged from 3674 to 11 236 mg kg�1 (Fig. 1). In the case of one of

the sludges (KR), there was a significant excess in the permissible

levels for biosolids intended for agricultural usage. In the case of

the other sludges these values were close to the permissible levels.

The PAH sum determined was relatively low when compared to

data presented by other authors.8,26,27 Mainly, 4-ring PAHs

predominated in all sludges. Among individual PAHs, the

highest share was noted for pyrene and fluoranthene. Moreover,

a high content of benzo[a]pyrene was found in sludge KR (Table

S1 and S2). Fluoranthene and pyrene are common PAHs

appearing in sewage sludges.7,28 The high concentration of

pyrene in the sewage sludges can be explained by the presence of

domestic sewage water in them. Charbroiling of meat accounts

This journal is ª The Royal Society of Chemistry 2008

Fig. 1 Total and potentially bioavailable PAHs content in sewage

sludges (A) and composts (B). Error bars represents standard deviation

error (SD, n ¼ 3 extractions).

Fig. 2 Changes of total PAHs content during composting.

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for a major source of these compounds, together with fluo-

ranthene, in households.9

Composting caused a significant decrease in the PAH sum in

the case of all sludges (Fig. 2). After 76 days of composting, the

total PAH content decreased in the range from 29.3 to 48.6%,

depending on the sewage sludge. The largest range of losses was

noted in sludges ZM and BJ, i.e. 48.6 and 41.0%, respectively.

The least favorable disappearance of the total PAHs content was

observed in sludge KR, as over 70.7% of PAHs were still present

in the composted material. PAHs removal observed during

composting may be the result of biodegradation of PAHs.

During the process of composting, the microorganisms capable

of degrading organic matter could be responsible for co-meta-

bolic degradation of the PAHs but similarly as in the soils,

pollutants can undergo sorption by the components of the

compost, e.g. in the stable particle structures such as humic

substances. PAHs may have been converted into bound

This journal is ª The Royal Society of Chemistry 2008

residues,28 and a part of them may have been sequestered into

inaccessible micro-sites within the compost matrix,29 as the above

mentioned processes limit total PAH extraction with organic

solvents. Information concerning the changes in the content of

individual PAHs has been presented elsewhere.6

The content of PAHs extracted by Tenax-TA

Depending on the sludge, the PAH content determined by

Tenax-TA adsorbent ranged from 890 to 5424 mg kg�1 (Fig. 1A).

The Tenax-TA extracted PAH content was in proportion to the

total content of these compounds (extracted with dichloro-

methane). The highest contribution of the potentially bioavail-

able fraction was observed for sludge KR, where it constituted

more than half (54%) of the PAHs determined by dichloro-

methane. In sludges ZM, BJ and LB, the share of this fraction

was clearly lower, i.e. 28.1, 31.0 and 38.3%, respectively. In the

case of individual PAHs, the share of the potentially bioavailable

fraction was clearly different and depended both on the type of

compound and sewage sludge. The highest content of the Tenax-

TA extracted fraction was noted for the 3-ring compounds

(mainly phenanthrene) which constituted from 77.0 to 84.5% of

the total content in individual sludges (Fig. 3A). The 4-ring

PAHs constituted another group with a share of 33.4–69.8%. The

highest content levels in this group was observed in the case of

fluoranthene in sludges ZM (80.9%) and BJ (81.9%), together

with pyrene in sludges KR (84.8%) and LB (78.6%).

However, in the case of most 4-ring PAHs, their share in the

Tenax-TA extracted fraction did not exceed 20%. The share of 5

and 6-ring PAHs was the lowest and ranged from 3.8 to 9.8% and

from 12.1 to 28.0%, respectively.

After composting, a significant decrease (P < 0.05) of the

Tenax-TA fraction content in the case of sludges KR, BJ and LB

was noted, by 83.8, 40.8 and 39.8%, respectively (Fig. 1B). Sludge

BJ was an exception. The content of the Tenax-TA fraction in

sewage sludge BJ did not undergo a significant change after the

composting process. There were clearly visible changes in the

share of individual PAHs as a result of composting. The scope

and direction of these changes varied for different sludges.

Composting of sludge KR resulted in a lowering of the share of

J. Environ. Monit., 2008, 10, 883–888 | 885

Fig. 3 Contribution of individual groups of PAHs in sewage sludges (A)

and composts (B). Error bars represents standard deviation error (n ¼ 3).

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most of the individual PAHs. In the case of sludges LB, ZM and

BJ, however, the share of the bioavailable fraction was higher

after composting for the majority of PAHs.

In the case of individual PAHs, a change in the content of the

Tenax-TA fraction varied and depended primarily on the sewage

sludge. In sludge KR, the scope of losses in the Tenax-TA

extracted fraction ranged from 28.1 to 98.4%. Chrysene was an

exception, as its content in the potentially bioavailable fraction

increased significantly. In sludges ZM, BJ and LB, more often

than in sludge KR, an increase in the content of the Tenax-TA

extracted fraction was noted after composting. Such an increase

was observed for anthracene, benzo[a]anthracene and benzo[a]

pyrene (ZM, BJ and LB), benzo[ghi]perylene (BJ, ZM), fluo-

ranthene and indeno[1,2,3-cd]pyrene (LB) (Table S1 and S2†).

The increase observed in the Tenax-TA fraction as a result

of composting can be related to the re-mobilization process

from the forms which used to be sequestrated or unavailable.

Re-mobilization of the sequestrated compounds was observed

earlier for sediments.30,31 Schlebaum et al.30 noted redistribution

of pentachlorobenzene from a slowly desorbing compartment to

the rapidly desorbing compartment after gas purging of the

sediment. The appearance of re-mobilization in the case of PAHs

was also noted by Kraaj et al.31 The phenomenon of pollutants

re-mobilization during composting was most probably related to

re-arrangement phenomena in the organic matter;32,33 PAHs

initially adsorbed to the sites which were temporarily less solvent

extractable.6 At the onset of composting, PAH molecules were

886 | J. Environ. Monit., 2008, 10, 883–888

entrapped in the closed interstitial spaces between small aggre-

gates or organic matter structures, which significantly limited

their bioaccessibility. As a result of organic matter changes

during composting, an opening in the closed interstitial spaces

may have taken place, hence resulting in an increase in the

content of pollutants over that being bioaccesible earlier.

The composting process has also significantly influenced the

share of the Tenax-TA fraction in the case of individual PAH

groups (Fig. 3B). However, the direction of these changes

depended on the compound and sewage sludge.

Both before and after composting, the share of the Tenax-TA

extracted fraction depended clearly on the log Koc (contaminant

sorption capacity to sewage sludges or compost matrices). With

an increase in log Koc of individual compounds, a decrease in the

share of the PAH fraction extracted with Tenax-TA was noted

(Fig. S1†). High weight molecular PAHs (5 and 6-ring

compounds) are characterised by higher hydrophobicity than 3

and 4-ring ones, which, as is generally known, causes an increase

of adsorption to organic matter34 and reduction of bioavail-

ability.35 The higher reduction of high weight molecular PAHs

results more from their strong binding by the sludge or compost

matrix than from biodegradation processes. Adsorption is less

reversible with increased numbers of condensed aromatic rings

or increased hydrophobicity.35

Prediction by Tenax-TA PAHs dissipation from composted

sewage sludges

The aim of the present study was to examine the feasibility of

using the Tenax-TA adsorbent for the evaluation of progress in

the losses of polycyclic aromatic hydrocarbons during sewage

sludge composting. Fig. 4 presents relationships between losses

of individual PAHs and the fraction of these compounds

extracted with Tenax-TA. The results obtained in this study were

satisfactory in the case of a small number of PAHs only. The best

reflection of the PAH losses was noted in the case of anthracene

in sludges ZM, BJ, LB; fluoranthene in sludges KR and LB and

indeno[1,2,3-cd]pyrene in sludges KR and ZM (Fig. 4). The

remaining satisfactory values concerned benzo[a]anthracene

(KR) and chrysene (ZM).

The method using the Tenax adsorbent is relatively popular,

and the results obtained with it satisfactory.16,17 It has been

shown previously36 that Tenax-TA extraction correlated very

well with biodegradation in natural (non-spiked) sediments,

however only when the sediments were extracted with several

changes of Tenax for up to 100 hours. Other studies20,37,38 showed

that satisfactory results are also obtained in a shorter period of

time (6–30 h). Despite that, the studies presented here give no

satisfactory result that would allow for the application of this

method in practice to composted sewage sludges. A prolongation

of contact time between Tenax-TA and sewage sludge could be

necessary to obtain satisfactory results. Losses of HMW PAHs

were larger than those extracted during 6 hours Tenax-TA

extraction. Difficulties in PAH loss forecasting are most prob-

ably related to specific sewage sludge properties, as sludges

constitute a very complex matrix.39–41 Sewage sludges differ

considerably both in their composition and content of individual

components that can exert a varying influence on the contami-

nant. Attention should also be drawn to surfactants, lipids and

This journal is ª The Royal Society of Chemistry 2008

Fig. 4 Correlations between disappearance of individual PAHs rate and

PAHs determined in the Tenax extracts of the fresh sludge. The dashed lines

on each plot are of a 1 : 1 slope. Black/filled point represents compounds with

good prediction (statistically significant) of their losses during composting.

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black carbon, all of which influence PAH sorption according to

numerous articles.42–44 During composting, some of the

contaminant–matrix bonds can become stronger whereas other

bonds can become weaker. Depending on the temperature during

This journal is ª The Royal Society of Chemistry 2008

composting, both the intensity and range of desorption can also

vary.43,45 Humidity is an important factor that can reduce/

increase hydrophobic compounds sorption.46 Such differentiated

composting conditions (differing in the case of individual

sludges) can make the effective and precise forecasting of

individual PAH losses considerably more difficult.

As shown in the studies presented by other authors,37,47

organic matter of sewage sludge is unstable and changes its

composition and properties during composting. During such

transformations, contaminants can be re-mobilised. However,

the fate of the ‘‘released’’ contaminant is not known. They may

enrich the bioavailable fraction (and undergo further degrada-

tion), as re-evaluations observed. On the other hand, they can

also undergo strong binding by the compost matrix. What is

more, full degradation of contaminants may be inhibited as

a result of other limitations.

It should be emphasised that in all sludges there were still

potentially bioavailable PAH forms present after 76 days of

composting (Tables S1 and S2†). Hence, it can be assumed that

the process of losing individual PAHs was not complete. The

presence of the bioavailable fraction excludes the idea that the

mass-transfer of chemicals was a limiting factor in further

pollutant degradation. Hence, there must be other reasons that

limit PAH degradation (e.g. oxygen deficit).

Conclusion

Both the content and contribution of potentially bioavailable

forms undergo changes as a result of composting. In the resulting

compost there are still ‘‘free’’ forms of pollutants (extracted with

Tenax). The appearance of such free forms in the compost

showed that PAH losses were not limited by the sequestration

processes. Other factors must limit PAHs losses during com-

posting. The instability of organic matter is most probably the

main factor making practical adaptation of the method tested

difficult. Changes in the formation taking place within the

organic matter during composting considerably influence the

interaction strength between pollutants and composted sewage

sludge matrix. Such a varying interaction strength at various

composting stages, as well as different process conditions, do not

allow for a precise determination of the bioavailable contami-

nant fraction during sewage sludges composting. Despite the

above mentioned difficulties, it is important to determine how

potentially bioavailable contaminant forms in the sludges or

composts would behave in the soil during its fertilization with

these sludges or composts.

Acknowledgements

The work was founded in the frame of grant No 2 P06S 005 29

financed in 2005–2008 from the budget of Ministry of Science

and Information Society Technologies.

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