REVIEW ARTICLE

y J. L. Berna, G. Cassani, C.-D. Hager, N. Rehman, I. López, D. Schowanek, J. Steber, K. Taeger and T. Wind

Anaerobic Biodegradation of Surfactants –

Scientific Review

The anaerobic biodegradation of surfactants is used as an ac- nen. Dieser wissenschaftliche Übersichtsartikel schlussfolgert,
ceptability criterion in some environmental pieces of legislation dass die anaerobe Abbaubarkeit nicht von gleicher Relevanz ein-
(eco-label, risk assessment, etc.), without a proper evaluation of zustufen ist wie die aerobe. Daher sollte die anaerobe Abbau-
the relevance of such a characteristic. Available screening test barkeit nicht als ein Entscheidungskriterium zur Umweltakzep-
methods to assess the anaerobic biodegradation do not simu- tanz von Tensiden genutzt werden, die unter aeroben
late the real conditions prevailing in these anaerobic compart- Bedingungen leicht biologisch abbaubar sind.
ments but rather reflect more stringent conditions, due to the
high test substance/biomass ratio, possibility of inhibitory ef- Stichwörter: Tenside, anaerober Abbau, anaerobe Abbaubar-
fects and limited possibility for adaptation. Therefore positive keit, Abwasserbehandlung, Screening-Tests, Simulationstests,
results are indicative of a similar behaviour under environmen- Sediment, Daten-Monitoring
tal conditions, while a negative result cannot be necessarily in-
terpreted as inherent anaerobic recalcitrance. The majority of
surfactants entering the environment will be exposed to and Table of Contents
degraded under aerobic conditions, and only less than 20 %
will potentially reach temporarily anaerobic environmental Foreword and Position Paper 313
compartments. In contrast to the well documented adverse ef-
1 Introduction 314
fects observed in the absence of aerobic biodegradation, the
lack of anaerobic biodegradation does not seem to be corre- 2 Executive Summary 314
lated with any apparent environmental problem for most com- Environmental relevance of anaerobic biodegrad-
partments after more than 40 years of widely use of such ability of surfactants
products. The scientific review concluded that anaerobic biode-
3 Surfactants in Europe Production/Consumption 317
gradability does not have the same environmental relevance as
the aerobic one. Anaerobic biodegradability should not, there- 4 Definitions 317
fore, be used as a pass/fail property for the environmental ac- Anaerobic biodegradation
ceptability of surfactants which are readily biodegradable under
5 Anaerobic Compartments 318
aerobic conditions.
5.1 Overview
5.2 Terrestrial
Key words: Surfactant, anaerobic biodegradation, anaerobic
5.3 Aquatic
biodegradability, wastewater treatment, screening tests, simula-
5.3.1 Water
tion tests, sediment, monitoring data
5.3.2 Sediment
5.3.3 ‘Extreme’ habitats
5.3.4 Groundwater
Anaerober Abbau von Tensiden – Wissenschaftliche Über-
5.4 Wastewater treatment
sicht. Der anaerobe Abbau von Tensiden wird häufig – ohne
5.5 Landfill
wissenschaftliche Bewertung der Relevanz einer solchen
5.6 Animals
Kenngröße – als ein Akzeptanzkriterium in einigen umweltrele-
vanten Teilen von Verordnungen (Ecolabel, Risikoabschätzun- 6 Existing Methods 319
gen, etc.) genutzt. Bekannte Screening-Testmethoden zur Ab- 6.1 Screening tests
schätzung des anaeroben Abbaus simulieren nicht reale 6.1.1 Anaerobic screening tests based on gas
Verhältnisse, die in anaeroben Kompartimenten vorherrschen, volume measurement only
sondern spiegeln aufgrund des hohen Testsubstanz/Biomasse- 6.1.2 Anaerobic screening tests based on gas
Verhältnisses sowie möglicher Inhibitionseffekte und begrenzt production measurement in the gas and
möglicher Adaption deutlich strengere Bedingungen wider. the liquid phase
Folglich weisen in Screening-Tests positive Abbauergebnisse auf 6.1.3 Predictive value of anaerobic biodegrada-
ein ähnliches Verhalten unter Umweltbedingungen hin, wäh- tion screening test data
rend ein negatives Ergebnis nicht unbedingt als inhärent anae- 6.2 Simulation tests
robe „Hartnäckigkeit“ zu interpretieren ist. Die Mehrzahl der in 6.2.1 Introduction to anaerobic simulation
die Umwelt eingetragenen Tenside wird aeroben Bedingungen tests
exponiert und unter diesen auch abgebaut. Nur weniger als 6.2.2 Test systems
20 % der Tenside erreichen eventuell temporär anaerobe Um- 6.2.2.1 14C-Anaerobic digester simulation
weltkompartimente. Im Gegensatz zu gut dokumentierten nega- test
tiven Effekten, die in Ermangelung von aerobem Abbau beo- 6.2.2.2 Anaerobic transformation test in
bachtet werden, scheint für die meisten Kompartimente ein aquatic sediment systems
mangelnder anaerober Abbau – auch nach mehr als 40 Jahren (OECD guideline 308)
weitem Gebrauch entsprechender Tenside – nicht mit irgendei- 6.2.2.3 Continuously operated anaerobic
nem ersichtlichen Umweltproblem assoziiert werden zu kön- reactors

312 ª Carl Hanser Publisher, Munich Tenside Surf. Det. 44 (2007) 6

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review
6.2.2.4 Biological nutrient removal STP
simulation tests: Behr and CAS-
UCT units
6.2.2.5 Others systems
7 Interpretation of Available Data on Anaerobic
Biodegradation of Surfactants 324
7.1 Anionics
7.1.1 Sulfonates
7.1.2 Sulfates
7.1.3 Fatty acids and soaps
7.2 Nonionics
7.3 Cationics – Amphoterics
8 Evaluation of the Relevance of Anaerobic Biodegra-
dation 334
8.1 Understanding mass fluxes of surfactants in
environmental compartments
8.1.1 Mass fluxes obtained via modelling for
hypothetical surfactants
8.1.2 Mass fluxes based on monitoring data
8.2 Impact of surfactants on structure and func-
tion of anaerobic environmental compart-
ments
8.2.1 Speciation and bioavailability of surfac-
tants in anaerobic compartments
8.2.2 Impact on wastewater treatment and
aquatic environments
8.2.2.1 LAS and other anionic surfactants
in anaerobic digesters
8.2.2.2 Other surfactants in anaerobic di-
gesters
8.2.2.3 Septic tanks and decentralized
treatment systems
8.2.2.4 Anaerobic river and lake sedi-
ments
8.2.2.5 Agricultural soil (pasture and
cropland)
8.2.2.6 Landfills for sludge
8.2.2.7 Marine sediments
References 340
Glossary Of Abbreviations 343
Appendix 344
Overview of surfactant monitoring and mass balan-
cing in anaerobic environmental compartments
Foreword to Revision 0 (1999)
This document was commissioned by ERASM (Environ-
mental Risk Assessment Steering Committee – a detergent
industry group).
ERASM was created in 1991 as a forum for the co-ordi-
nation of views and actions in the field of risk assessment
and risk assessment issues between the chemical industry
(CESIO – Comit
Europ
en des Agents de Surface et leurs
Interm
diaires Organiques, a CEFIC Sector Group) and the
detergent industry (AISE, Association Internationale de la
Savonnerie, de la D
tergence et des Produits d’Entretien).
ERASM’s mission is to define the industry position in
establishing and managing the risk of surfactants to the en-
vironment and to propose reasonable and technically sound
guidelines for assessing the environmental risk of surfac-
tants. ERASM also ensures that resources are available to
carry out necessary research programmes.
The structure of ERASM consists of ad hoc task forces
established to work on specific topics related to risk assess-
Tenside Surf. Det. 44 (2007) 6
ment methods and data. A task force was set up to address
anaerobic biodegradation and the environmental relevance
of anaerobic biodegradability of surfactants.
The present dossier gives a compilation and interpreta-
tion of the available literature on the fate and biodegradabil-
ity of commercial surfactants under anaerobic conditions.
The members of this specific task force are:
José Luis Berna (Chairman) Petresa (Es)
Nigel Battersby Shell Chemicals (UK)
Luciano Cavalli Condea Augusta (It)
Richard Fletcher Unilever (UK)
Andreas Guldner BASF (De)
Diederik Schowanek Procter & Gamble (Be)
Josef Steber Henkel (De)
Foreword to Revision 1 (2007)
The document published in 1999 has been updated with
new data published in the scientific literature in the period
1999 – 2005. The new dossier represents therefore the most
recent compilation of data, interpretation and assessment
of the relevance of anaerobic biodegradation of surfactants.
The report also contains data referring to surfactants
which are not considered as relevant in the detergent indus-
try today. They have been included however with the pur-
pose of having a comprehensive data base.
The members of the ERASM task force responsible for
this update are:
y Jos
Luis Berna (Chairman) Petresa (Spain)
y Giorgio Cassani Sasol (Italy)
y Claus-Dierk Hager Sasol (Germany)
y Ignacio Lpez Serrano Petresa (Spain)
y Diederik Schowanek Procter and Gamble
(Belgium)
y Josef Steber Henkel (Germany)
y Thorsten Wind Henkel (Germany)
y Klaus Taeger BASF (Germany)
y Naheed Rehman Unilever (UK)
Position Paper
The anaerobic biodegradation of surfactants is used as an ac-
ceptability criterion in some environmental pieces of legisla-
tion (eco-label, risk assessment, etc.), without a proper eval-
uation of the relevance of such a characteristic.
Surfactants form a group of chemicals with considerable
environmental importance due to their high volume con-
sumption and widespread use as they are essential ingredi-
ents in most laundry and cleaning products. Since the major
part of the biosphere is aerobic, priority has been given to
the study and assessment of biodegradability under these
conditions. Nevertheless there are environmental compart-
ments which can be permanently (e. g. anaerobic digesters)
or temporarily anaerobic (e. g. river sediments and soils) and
surfactants do reach these.
Available screening test methods to assess anaerobic bio-
degradation do not simulate the real conditions prevailing in
these anaerobic compartments but rather reflect more strin-
gent conditions, due to the high test substance/biomas ratio,
possibility of inhibitory effects and limited possibility for
adaptation. Therefore positive results are indicative of a sim-
ilar behaviour under environmental conditions, while a ne-
gative result cannot be necessarily interpreted as inherent
anaerobic recalcitrance. In addition, low biodegradation re-
sults in these tests may be influenced by a limited bioavail-
ability due to the formation of insoluble chemical species.
313
J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review
The relevance of anaerobic biodegradability cannot be
separated from other important properties of surfactants
such as sorptive behaviour, ecotoxicity profile and above all,
aerobic biodegradation rate.
The majority of surfactants entering the environment
will be exposed to and degraded under aerobic conditions,
and only less than 20 % will potentially reach anaerobic, en-
vironmental compartments. In all but a few cases their pre-
sence in these will not be permanent.
A systematic evaluation of the risk to the structure and
function of these compartments due to the presence of
non-degraded surfactants led to the conclusion that, in con-
trast to the adverse effects observed in the absence of aerobic
biodegradation, the lack of anaerobic biodegradation does
not seem to be correlated with any apparent environmental
problem for most compartments. Particularly for the sedi-
ment compartment, data is lacking and it is recommended
to fill the missing data gaps to assess structure and function.
In criteria for eco-labelling a conservative set of “scoring”
or “weighting” factors, if any, for anaerobic biodegradability,
should follow from a combination of the above characteris-
tics, and it is suggested that these should be of the order of
one tenth of the aerobic biodegradability value for readily
biodegradable surfactant.
Consequently it is concluded that anaerobic biodegrad-
ability does not have the same environmental relevance as
the aerobic one. Anaerobic biodegradability should not,
therefore, be used as a pass/fail property for the environ-
mental acceptability of surfactants which are readily biode-
gradable under aerobic conditions.
1 Introduction
Biodegradation is the most important mechanism for the
irreversible removal of chemicals from the aquatic and ter-
restrial environments. Therefore, the evaluation of biode-
gradability is an indispensable element of the exposure
assessment within the environmental risk assessment pro-
cess of a chemical substance.
The biosphere is predominantly aerobic and it is under-
standable that the biodegradation behaviour of chemicals
under aerobic conditions has been the focus of attention
for a long time. This has led to the development of a num-
ber of internationally used and recognised laboratory meth-
ods for assessing aerobic biodegradability and to a huge
amount of test data. The biodegradability of chemicals in
the absence of free oxygen, i. e., under anaerobic conditions
has been considered to a far lesser extent. Nevertheless,
there are environmental areas which are permanently or
temporarily anaerobic (e. g. sludge digesters of sewage treat-
ment plants, sediments or sub-surface soil layers). It can be
argued therefore that a comprehensive evaluation of a chem-
ical’s environmental fate should also address its anaerobic
biodegradability, provided that there is a possibility for enter-
ing anaerobic environments to a significant extent.
Typically, such substances are characterised by a poor
water solubility or strong adsorptivity onto solids, resulting
in an environmental distribution with a pronounced trans-
portation into anaerobic compartments. Surfactants form a
group of chemicals with a high environmental relevance
and physical-chemical properties which may result in a sig-
nificant partitioning between the aqueous and the solid
phase in the aquatic environment. Environmental monitor-
ing data for some surfactants support this partitioning as-
sumption, showing their presence in secondary and digester
sludges, and sediments. The same substance properties are
also relevant in the context of the environmental risk assess-
314
ment of chemicals in the soil compartment as application of
digested sludges for agricultural purposes will influence the
initial environmental concentration in soils.
In order to establish an overview of the anaerobic biode-
gradation aspects in general (i. e. environmental relevance,
test methodology) and of their relevance for surfactants in
particular, an ERASM Task Force was created. This position
paper was prepared by experts of the involved industries and
aims to form a common basis of knowledge on this topic.
2 Executive Summary
Environmental relevance of anaerobic biodegradability
of surfactants
Aim of the review
The objective of this review, prepared by the ERASM Task
Force ‘Anaerobic Biodegradability’ was to provide a thorough
compilation and interpretation of the available literature on
the fate and biodegradability of commercial surfactants un-
der anaerobic conditions. This analysis should form the ba-
sis for a data-based discussion on the environmental rele-
vance of the property of anaerobic biodegradability (or lack
thereof) in a broad range of contexts, such as risk assess-
ment of chemicals, the upcoming review of the Detergents
Regulation (EC) 907/2004 (addressing specifically the anae-
robic biodegradability of detergent surfactants, the biode-
gradability of non surfactant organic ingredients and the
use of phosphates) and the eco-labelling of detergent pro-
ducts.
The earth’s atmosphere contains just over 20 % of oxygen
by volume making aerobic compartments the predominant
ones in the biosphere. It is therefore not surprising that the
prevalent route for biodegradation of chemicals is aerobic
and, hence, aerobic biodegradation has been extensively
studied. In addition, it is a legal requirement in the Eu-
ropean Union (Detergent Regulation) for surfactants used
in detergents to be readily (and ultimately) biodegradable
under aerobic conditions.
Surface active agents are essential ingredients in most
household laundry products, domestic and industrial clean-
ers, as well as in personal care, cosmetic products and a vari-
ety of industrial applications. Surfactants form a group of
chemicals with a high overall environmental relevance, due
to a combination of their inherent environmental properties
and their very large production volume. They are typically
discharged into the environment through the sewage treat-
ment infrastructure (i. e. sewers, sewage treatment plants),
or directly in situations where no treatment systems are
available.
Anaerobic test methods and their predictive value
Test methods to determine the ultimate anaerobic biode-
gradability of organic compounds at screening and simula-
tion level are available. First, there are screening tests such
as the ECETOC test/ISO 11 734/OECD-311 which deter-
mine anaerobic mineralization by measurement of the
methane and carbon dioxide gas production via a pressure
or volumetric reading and DIC measurement. Due to the
stringent screening-type test conditions (i. e. high test sub-
stance/biomass ratio), inhibitory effects of surfactants can-
not be excluded and the possibilities for acclimation are lim-
ited. These tests are reliable at avoiding false-positive
conclusions, but a poor result is not necessarily proof of
anaerobic recalcitrance. In simulation tests the substance/
biomass ratio is much lower and can reach real conditions
Tenside Surf. Det. 44 (2007) 6
J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review
(e. g. 8,000 mg C/kg dry solids, if gas production is evalu-
ated, or even lower, if specific analytics is applied).
Anaerobic biodegradation test systems under more rea-
listic conditions (in terms of sludge concentration, fresh
sludge feed, residence times, etc.) include digester simula-
tion tests employing radio labelled test substances or discon-
tinuous systems. Limitations, however, remain for situations
such as low substrate concentrations and toxic surfactants.
While, in principle, test methods for simulation of aerobic/
anaerobic river sediments exist, experimental data based on
these improved testing methodologies are scarce. Also, the
application of specific analytical methods to follow the fate
of surfactants and their breakdown products in anaerobic
matrices is not widespread as yet.
Anaerobic biodegradability of surfactant groups
Based on the aforementioned laboratory test methods, the
available data from published biodegradation studies and
available monitoring studies allow the evaluation of the
anaerobic ultimate biodegradability of the important surfac-
tant groups used in detergents:
y Sulfonated anionic surfactants (LAS, SAS, MES, AOS):
poorly biodegradable
y Sulfated anionic surfactants (AS, AES): well biodegrad-
able
y Fatty acids and soaps: well biodegradable
y Alcohol ethoxylates (AE): well biodegradable
y Sugar-based non-ionic surfactants (APG, glucamides):
well biodegradable
y Alkylethanolamides: well biodegradable (based on lim-
ited data)
y Alkyldimethyl amine oxides: well biodegradable (based
on limited data).
y APEOs*: partially degradable leaving alkylphenol resi-
dues.
y Mono- or di-alkyl quaternary compounds (TMAC,
DTDMAC*): poorly biodegradable.
y Esterified mono- or di-alkyl quaternary surfactants (ester-
quats): well biodegradable.
y Amphoteric surfactants (alkylamido betaines, alkylbe-
taines, alkylamphoacetates): well biodegradable (based
on limited data)
* Not relevant anymore in detergent applications
Factors for evaluating the importance of anaerobic biodegrada-
tion
Mass flux to anaerobic environments: Strictly aerobic and
anaerobic environments represent the two extremes of a
continuous spectrum of environmental habitats which are
populated by a broad variety of micro-organisms with specif-
ic biodegradative capabilities. Due to the specific usage pat-
terns, the bulk of the surfactant mass entering the environ-
ment will predominantly be exposed to and degraded under
aerobic conditions. For readily biodegradable surfactants,
less than 1 % of the flux will reside in the permanently anae-
robic compartments of a landfill and the occasionally anae-
robic compartment of agricultural land.
The relative importance of sewage treatment has in-
creased considerably during the last decade with a higher
treatment incidence, halting of sludge disposal into the sea
and improved treatment systems with biological nutrient re-
moval. Incineration of waste water sludge has become a
more important disposal route. Consequently, the amount
of surfactants which can reach anaerobic compartments via
sludge disposal has decreased and is likely to decrease
316
further over time (Urban Wastewater Treatment Directive
91/271). In spite of the EU Directive 91/271 generally still
the quality of wastewater treatment is very different in the
EU.
The differences in physico-chemical properties and bio-
degradability explain the observed differences between the
mass fluxes of different surfactants. As such, the relevance
of anaerobic biodegradability cannot be separated from
other important properties of the surfactants, such as sorp-
tive behaviour (log Kow), solubility, and most importantly,
aerobic biodegradation kinetics.
Accordingly, a German Expert Group for Detergents
(HAD)1 had previously proposed ranking the relevance of
anaerobic biodegradability as a function of the above proper-
ties.
The recently conducted environmental risk assessments
of all important surfactants used in detergents (HERA) un-
derline the considerably higher environmental relevance of
the aerobic biodegradability. Recently (Nov. 2005), SCHER
opinion on the “Environmental Risk Assessment of non
Biodegradable Detergent Surfactants under Anaerobic Con-
ditions” has concluded that the requirement for ready and ulti-
mate anaerobic biodegradability under anaerobic conditions is
not by itself regarded as an effective measure for environmental
protection. These (aerobically) readily biodegradable sub-
stances proved to be of no concern for the aquatic and ter-
restrial environment regardless of their anaerobic biodegrad-
ability.
Bioavailability:
Changes of bioavailability of surfactants under anaerobic
conditions can affect the outcome of toxicity/inhibition stud-
ies, and to some extent biodegradation or removal rates. The
specific chemical structure of some surfactants contributes
to a rapid precipitation with water hardness ions (Ca++,
Mg++) into insoluble forms, as well as adsorption to the sur-
rounding solid matrix. This highlights the need to use the
real environmental form of a surfactant in inhibition and
biodegradation tests, in order to obtain a realistic test
result.
Impact of surfactants on the structure and function of anaerobic
compartments:
Based on available data, an evaluation was made of the risk
to the structure and function of anaerobic compartments
being affected by fluxes of non-degraded surfactants enter-
ing the system. The evaluation led to the conclusion that in
contrast to the adverse effects and high risk in the absence
of aerobic biodegradation, the lack of anaerobic biodegrada-
tion does not seem to be correlated with any apparent envir-
onmental or technical problems in most compartments.
Nevertheless, it is fair to say that for natural anaerobic envir-
onments, not all aspects of structure and function can be
adequately assessed today since comprehensive data are
lacking. Because of the precautionary principle knowledge
of the anaerobic degradability of surfactants would be pre-
cious. It is acknowledged therefore that surfactants biode-
gradable under both aerobic and anaerobic conditions leave
less room open to environmental issues.
Conclusions
Anaerobic biodegradability as a strict pass/fail criterion is
not in line with the environmental interpretation and signif-
1
Schöberl P. (1994). Die Bedeutung fehlender anaerober biologischer Abbau-
barkeit. Tenside Surf. Det. 31, 157 – 162.
Tenside Surf. Det. 44 (2007) 6
J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review

icance that should be given to the lack of this property for

surfactants. For surfactants in detergents, rapid aerobic bio-
degradation as well as their sorptive and ecotoxicological
properties are keys for a realistic environmental compatibil-
ity assessment. If a surfactant is rapidly degradable under
aerobic conditions, and its (transitory) presence in anaerobic
environments does not affect the function and structure of
that compartment (e. g. it is not inhibitory), then its anaero-
bic degradability is of minor importance. Nevertheless, it is
fair to say that for natural anaerobic environments, not all
aspects of structure and function can be adequately assessed
today since comprehensive data is lacking.

3 Surfactants in Europe Production/Consumption

Surfactants (anionics, nonionics, cationics and amphoterics)

are used in different fields such as cosmetics, metal work- Figure 1 Surfactant end uses in W. Europe
ing, mining, agriculture, paper and leather industries and
obviously the detergent industries.
CESIO compiles annual statistics on the production and
4 Definitions
consumption of surfactants in Europe.
The total surfactant market in Western Europe is mature Anaerobic biodegradation
and annual growth rates are expected to be 1 – 1.5 %.
The largest market for surfactants is in household appli- Anaerobic biodegradation means the microbial degradation
cations, which accounts already for half of the total surfac- of organic substances in the absence of free oxygen (O2).
tant consumption. In this market a limited number of sur- While oxygen serves as the electron acceptor in aerobic biode-
factant types remain dominant: alkylbenzene sulfonates, gradation processes forming water (H2O) as the final product,
alcohol-sulfates and alcohol ether sulfates, alkane sulfonates degradation processes in anaerobic systems depend on alter-
as well as fatty alcohol ethoxylates. Other outlets for surfac- native acceptors such as sulfate, nitrate or carbonate yielding,
tants are I&I (Industrial and Institutional cleaning), perso- in the end, hydrogen sulphide (H2S), molecular nitrogen (N2)
nal care, textiles, emulsion polymerization, paint additives, and/or ammonia (NH3) and methane (CH4), respectively.
agrochemicals, etc. where besides the a.m. commodity type Anaerobic biodegradation is a multistep process per-
surfactants, mainly “specialty” surfactants are used. Figure 1 formed by different bacterial groups. It involves hydrolysis
illustrates the wide field of applications of surfactants. of polymeric substances like proteins or carbohydrates to
monomers and the subsequent decomposition to soluble
acids, alcohols, molecular hydrogen (H2) and carbon dioxide
(CO2). Depending on the prevailing environmental condi-
Surfactants consumption tions, the final steps of ultimate anaerobic biodegradation
in Western Europe are performed by denitrifying, sulfate-reducing or methano-
(incl. captive use consumption)
genic bacteria (Figure 2).
in 1000 tonnes – as 100 % active 1995 2000 2005 In contrast to the strictly anaerobic sulfate-reducing and
A. ANIONICS methanogenic bacteria, the nitrate-reducing micro-organ-
isms as well as many other decomposing bacteria are mostly
Alkylbenzene sulfonates 410 450 490 facultative anaerobic (i. e. they are able to grow and to de-
Alkane sulfonates 74 72 64 grade organic substances under aerobic as well as anaerobic
conditions). Thus, strictly aerobic and anaerobic environ-
Alcohol sulfates 94 86 60
ments represent the two extremes of a continuous spectrum
Alcohol ether sulfates 192 261 400 of environmental habitats which are populated by a broad
Other anionics 80 75 90
Total Anionics (excl. soaps) 850 944 1104
Soaps 550 650 700
B. NONIONICS
Alkylphenol ethoxylates 82 86 30
Alcohol ethoxylates 440 470 530
Other ethoxylates 100 124 200
Other nonionics 170 163 120
Total Nonionics 792 843 880
C. CATIONICS 181 166 200
D. AMPHOTERICS 38 45 76
GRAND TOTAL 1861 1998 2260

Table 1 CESIO Statistics Figure 2 Anaerobic degradation processes by bacterial consortia

Tenside Surf. Det. 44 (2007) 6 317

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review
variety of micro-organisms with specific biodegradation abil-
ities. Anaerobic conditions occur where vigorous decomposi-
tion of organic matter and restricted aeration result in the
depletion of oxygen. Anoxic conditions may represent an in-
termediate stage where oxygen supply is limited, still allow-
ing a slow (aerobic) degradation of organic compounds. In a
digester the various bacteria also have different require-
ments to the surrounding environment. Acidogenic bacteria
need pH values from 4 to 6, whilst methanogenic bacteria
from 7 to 7.5 (Temper and Pfeiffer, 1986). In batch tests the
dynamic equilibrium is often interrupted because of an en-
richment of acidogenic bacteria as a consequence of lacking
substrate in- and outflow.
5 Anaerobic Compartments
5.1 Overview
The biosphere is predominantly aerobic with an estimated
1015 tons O2 being present in the atmosphere and oceans
(Press and Siever, 1986). However, anaerobic environments
can develop where the consumption of O2 by the biological
oxidation of organic matter exceeds supply. These may be
small anaerobic zones in an otherwise oxic system (e. g. Jør-
gensen, 1977a), or much larger and stable environments
such as those found in marine and freshwater sediments.
As was mentioned in Chapter 4, in the absence of free O2
the oxidation of organic matter by micro-organisms con-
tinues through the use of a sequence of alternative electron
acceptors (e. g. nitrate sulfate carbon dioxide). These reac-
tions are dependent on the availability of organic and inor-
ganic substrates, the redox potential of the environment
and the types of bacteria present (Zehnder and Stumm,
1988). This is illustrated in Figure 3 and shows a progres-
sion down idealised coastal sediment. It can be seen that
the higher the redox potential of the environment, the more
energetically favourable is the reaction. Hence, a facultative
anaerobic bacterium existing in an anaerobic zone with a
low redox potential, will quickly become more energy effi-
cient once there is an ingress of O2. This sequence of bacter-
ia preferring reactions which provide the most energy yield
can be found in a wide range of aquatic and terrestrial envir-
onments.
Anaerobic processes play an important role in nutrient
cycling and animal nutrition, and in man-made activities
such as wastewater treatment. This chapter summarises
the main anaerobic environmental compartments.
Figure 3 Simplified diagram of the different reactions performed by bacteria
depending on the redox potential of the environment. This example is for
coastal marine sediment and is not to scale
318
5.2 Terrestrial
Soils are usually aerobic systems (Kaspar and Tiedje, 1982),
although anaerobic micro sites can occur in poorly drained
soils and a flooded soil such as a rice paddy may have an
aerobic layer of only 1 cm depth (Richards, 1987). Another
important terrestrial anaerobic environment is the peat in
water-logged moor land, fens and bogs.
5.3 Aquatic
5.3.1 Water
An anaerobic body of water can occur if there is high O2
consumption in the sediment or overlying water, and seaso-
nal or permanent hydrographic conditions such as thermo-
clines or haloclines prevent the exchange of water. Many
freshwater lakes of > 10 m depth can become stratified at
the end of the summer, leading to the formation of an anae-
robic layer above the sediment (Jones, 1982).
The largest anaerobic ecosystems in the biosphere are
marine (Schlegel and Jannasch, 1981), with the Black Sea
being anaerobic from a depth of * 200 m to the bottom at
* 2,000 m. Other permanently anaerobic basins are the Car-
iaco Trench, off the coast of Venezuela (Richards, 1975) and
the Orca Basin in the northern Gulf of Mexico (Shokes et al.,
1977). These reports and others have led Sieburth (1987) to
conclude that anaerobic basins are fairly common. However,
the decomposition of organic material in these regions ap-
pears to occur predominantly in the oxygenated water col-
umn, with reports of * 75 % of organic matter being aerobi-
cally oxidised in stratified lakes (Rudd and Hamilton, 1979;
Harrits and Hanson, 1980) and 85 – 95 % in marine systems
such as the Black Sea (Deuser, 1971).
5.3.2 Sediment
Depending on the level of eutrophication, water depth and
season, freshwater sediments are usually anaerobic below
the surface few mm or cm. In general, aerobic degradation
is the predominant decomposition process in these sedi-
ments, although anaerobic processes such as dissimilatory
nitrate reduction and methane evolution are also important
(Jones, 1982).
The aerobic zone of marine sediments can vary from only
a few mm in coastal areas to ‡ 1 m in deep sea sediments (Jør-
gensen, 1982). The depth of this zone is dependent on factors
such as the rate of O2 diffusion into the sediment porewater,
bioturbation, microbial respiration rates (which in turn are
dependent on organic carbon levels), sediment particle size
and tidal flushing. Within the anaerobic layers, sulfate reduc-
tion is the predominant terminal step in anaerobic biodegra-
dation and this reflects the abundance of sulfate in sea water
(29 mM at a salinity of 35 %). In coastal sediments, the
mineralization of organic material by sulfate reduction is sig-
nificant and has been reported to match or even exceed that
due to aerobic respiration in intertidal salt marshes and mud-
flats (Howarth and Teal, 1979; Mountfort et al., 1980) and fjor-
dic sediments (Jørgensen, 1977b). However, these shelf (i. e.
shoreline to 200 m depth) sediments cover only around 9 %
of the world’s oceans (Jørgensen, 1982) although they receive
a disproportionately large amount of anthropogenic material.
5.3.3 ‘Extreme’ habitats
These are anaerobic environments which have extreme con-
ditions of temperature (> 100
C), salinity (saturated NaCl) or
Tenside Surf. Det. 44 (2007) 6
J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review
pH (< 2, > 10) and include geothermal vents, hot springs,
salt lakes, peat bogs and alkaline hypersaline waters (see
Lowe et al., 1993 for a detailed review). The release of surfac-
tants into these habitats is virtually non-existent and as such
they are ignored in this document.
5.3.4 Groundwater
Groundwater can become anaerobic, particularly when a
shallow aquifer is contaminated with degradable organic
material from a landfill leachate (Smolenski and Suflita,
1987). In deeper aquifers groundwater also can become
anaerobic because of natural percolation of rain water and
microbial activities. CO2 is absorbed during this passage
and Mn2+ and Fe2+ are dissolved. Therefore not only con-
tamination through organic material may lead to anaerobic
conditions (Bavarian Environment Agency).
5.4 Wastewater treatment
Mesophilic (i. e. * 35
C) anaerobic digestion of sewage
sludge is widely used in wastewater treatment to reduce vol-
ume, stabilise the sludge and produce CH4-rich biogas
which can be burnt to generate energy. Sludge digestion is
a strictly anaerobic process, with a redox potential of the or-
der of –250 mV (Mosey, 1985). Digester capacities usually
range from 1,000 to 10,000 m3 (Mosey, 1983) and the aver-
age residence time is 15 to 20 days. Anaerobic digestion
has also been promoted as a means of supporting the fuel
demands of developing countries, and large numbers of di-
gesters have been installed in China and India (Compag-
nion and Nyns, 1986).
Surfactants tend to adsorb onto sludge and as such are
passed into anaerobic digesters (Field et al., 1995). This is
possibly the anaerobic environment with the highest expo-
sure to surfactants (see also the monitoring summary table
in the Appendix). In a number of European countries, the
digested sludge is eventually disposed to land as a soil
amendment, whereby the plant and soil life may be exposed
to surfactants or their metabolites.
Other anaerobic environments associated with waste-
water treatment are: denitrifying filters and activated sludge
systems (Mosey, 1983), anaerobic ponds for high strength
industrial wastewaters (Mara and Pearson, 1986), primitive
treatment plants based on septic tanks where wastewater is
percolated, and anaerobic/anoxic zones in wastewater treat-
ment plants with biological removal (see below).
An increasing number of modern wastewater treatment
plants are fitted with anaerobic/low dissolved O2 zones in
which enhanced nutrient (phosphorous, nitrogen) removal
takes place. Tertiary wastewater treatment partly including
such biological nutrient removal processes covers around
75 % of the population connected to wastewater treatment
plants in Northern Europe, ca. 64 % in Central Europe and
< 10 % in Southern Europe and the EU Accession Countries
(EEA, 2002)2. The mentioned anaerobic/low dissolved O2
zones are usually situated prior to the aerobic zone and as
such surfactants may be subject to anaerobic/low dissolved
O2 attack prior to their aerobic biodegradation. The resi-
dence time of the mixed liquor suspended solids in both
the anaerobic and the low dissolved O2 tank is limited to
only a few hours. Anaerobic degradation steps can be
brought about by the fraction of the biomass active in these
2
EEA (European Environment Agency). Urban wastewater treatment. ETC/WTR
based on Member States data reported to OECD/EUROSTAT Joint Question-
naire 2002. (http://themes.eea.europa.eu/Specific_media/water/indicators/
WEU16, 2004.05)
Tenside Surf. Det. 44 (2007) 6
zones, which are mainly facultative anaerobic micro-organ-
isms. Fermentation reactions will take place, although the
final conversion to methane is not significant due to the lim-
ited residence time in the anaerobic zones. Redox conditions
vary considerably over time and place in the system. Due to
the longer hydraulic and sludge residence times in such sys-
tems and the diversity of degradation (redox) conditions, it is
expected that surfactant removal will generally be enhanced
in comparison to classical activated sludge plants (Rottiers
et al., 1998).
5.5 Landfill
Decomposition of organic material in landfills, particularly
those with compacted deposits, leads to the consumption of
O2 and the formation of anaerobic conditions (Kster and
Niese, 1986).
5.6 Animals
Anaerobic environments exist in the alimentary tracts of
many animals, in particular the rumen of cattle and the
blind sacs (ceca) of termites, wood roaches, rabbits, rats
and pigs (Latham, 1979). The fate of surfactants in these en-
vironments is beyond the scope of this paper.
6 Existing Methods
As with aerobic biodegradation test systems, a hierarchy of
screening, inherent and simulation tests has been proposed
for anaerobic conditions. Screening tests are characterised
primarily by a high test substance to biomass ratio, while in-
herent and simulation tests aim to reach realistic concentra-
tion ranges of the chemical and the bacterial biomass. In ad-
dition, screening and inherent tests usually have a relatively
simple test design (e. g. batch tests, which are not capable to
represent all the dynamic processes of anaerobic degrada-
tion) making them suitable for routine testing, whereas si-
mulation tests necessitate the use of 14C labelled materials
or specific analytical methods.
6.1 Screening tests
The available anaerobic biodegradation screening tests are
based on the determination of the final gaseous products of
the anaerobic degradation process under methanogenic con-
ditions, i. e. carbon dioxide (CO2) and methane (CH4).
However, the test conditions differ considerably from the si-
tuation in a real digester. The diluted sludge inoculum cor-
responds to about 10 % or less of the real digester sludge
concentration. In addition, for analytical reasons, the test
compound concentration is usually in the range of 20 –
100 mg of organic carbon/l, and is significantly higher than
the concentrations usually found in digesters. Therefore, in
some cases inhibitory effects are to be expected and also
have been observed in these screening tests. This has to be
taken into account when negative or poor results are ob-
tained in the test. From these facts it is understandable that
anaerobic screening tests are more stringent than test sys-
tems simulating realistic environmental conditions. As in
aerobic screening tests, it can be concluded that a positive
test result in an anaerobic screening test is indicative for
anaerobic biodegradability under real environmental condi-
tions whereas a negative result is not necessarily proof of re-
calcitrance.
319
J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review
6.1.1 Anaerobic screening tests based on gas volume measurement
only
A number of anaerobic biodegradability screening test pro-
cedures are based on the method described by Shelton and
Tiedje (1984), e. g. the test methods used in USA (ASTM,
1987), EPA (1988), the UK procedure (HMSO, 1989), Bat-
tersby and Wilson (1988) and the method described by Bau-
mann and Schefer (1990).
Test principle
Primary anaerobic digester sludge (with 1 – 3 % w/v dry sol-
ids) is diluted to 10 % with a mineral salt medium yielding a
sludge concentration in the test of 1 – 3 g of dry solids/l. The
test vessels (serum bottles) used have a nominal volume of
about 160 ml and contain 100 ml of the sludge mixture as
well as the test chemical at a concentration of 50 mg of or-
ganic carbon/l. After sealing, the bottles are incubated at
constant temperature (35 8C) in the dark for a test period
up to 8 weeks. The gas production is measured periodically
by determination of the pressure increase in the headspace
of the bottles using a calibrated pressure meter. The net
amount of gas produced from the degradation of the test
substance (test gas production corrected for inoculum blank
gas production) is expressed as a percentage of the theoreti-
cal gas production (ThGP) calculated from the chemical for-
mula of the substance taking into account the theoretical
ratio of CO2 and CH4 formed in the digesting process (Bus-
well equation) and the empirical solubilities of the gases in
the test medium.
Buswell equation:
CnHaOb + (n – a/4 – b/2) H2O
fi (n/2 – a/8 + b/4) CO2 + (n/2 + a/8 – b/4)
Technical aspects
The method is applicable to water soluble and poorly soluble
substances provided the concentration of the test material
(50 mg of carbon/l) is not inhibitory to the anaerobic organ-
isms. Knowledge of the chemical structure or, at least, of the
empirical formula of the test compound is necessary so that
the theoretical maximum amount of the gaseous final pro-
ducts (CO2 and CH4) can be predicted employing the Bus-
well equation. Since the test procedure only measures the
headspace gas pressure and volume, respectively, it is neces-
sary to make assumptions about the relative solubility of the
gases in the test mixture.
The advantage of relatively simple technical require-
ments for conducting the test faces a few drawbacks:
i) If the chemical formula of the test material is unknown,
the Buswell equation cannot be applied and, thus, the
amounts of the individual gases cannot be calculated.
ii) The ratio of CO2 and CH4 evolved in the test may differ
more or less considerably from the Buswell equation as
was shown in experiments with radio labelled test sub-
stances (ECETOC 1988). If the ratio cannot be predicted
reliably, the amounts of the two gases in solution cannot
be calculated.
iii) The true amount of dissolved CO2 may be variable since
the solubility of this gas depends on a number of factors
(e. g., temperature, pressure, pH, ratio of headspace/liq-
uid volume, thermodynamic equilibrium established be-
tween CO2 and carbonates/bicarbonates of Ca and Mg)
not measured in the test. For that reason, the prescribed
test conditions have to be rigidly adhered to (Shelton and
Tiedje, 1984; Battersby and Wilson, 1988).
320
Nevertheless, in spite of these problems the method is
widely applicable and considered to be a reasonably accurate
screening procedure for the evaluation of the anaerobic de-
gradation of test materials.
Test modifications
The method described by Baumann & Schefer (1990) differs
somewhat from the previously described procedure by using
an extended Buswell equation to calculate directly the CH4
production from the chemical oxygen demand of the sub-
stance or product tested. In addition, a lower inoculum con-
centration (0.5 g/l) is used and the problem of the solubility
of CO2 is overcome by adding NaOH to the digesting mix-
ture when gas production has reached a plateau. The test
flasks, fitted with stirrers, are larger than in the afore-men-
tioned method (250 ml nominal volume containing 200 ml
of liquid) and the gas production is measured by means of
a mercury manometer fitted to each flask.
6.1.2 Anaerobic screening tests based on gas production measurement
in the gas and the liquid phase
The basic pertinent test system is the so-called ECETOC
(1988) test developed and published by the European Centre
for Ecotoxicology and Toxicology of Chemicals. The method
has been ring-tested and is standardised as ISO 11 734
method (ISO 1995)”. The OECD 311 test guideline (2003)
is an adaptation of ISO 11 734. The principle on which the
ECETOC test and the mentioned standard test methods are
based upon is shown in Figure 4.
ISO/DIS 14 853 (1999) represents another draft standard
which is essentially identical with ISO 11 734 but offers, in
addition to the manometric method, a volumetric measure-
ment of the biogas production.
Test principle
A known volume of washed anaerobic sludge (1 – 3 g/l total
solids), suspended in an oxygen-free mineral medium, is
placed in a suitable vessel (nominal volume 0.1 – 1 l) leaving
a headspace (10 – 40 % of the volume of the vessel) into
which any gases produced can evolve. Prior to sealing, a
small amount of the test compound is added to give a con-
centration of 20 – 100 mg of organic carbon/l. Controls
(without test compound) are prepared in the same manner.
Usually the test is carried out using several (e. g. 5) replicates
each of controls and test assays. The vessels are incubated at
constant temperature (35 € 2 8C) normally for periods of up
to 60 days. The headspace pressure resulting from the pro-
duction of gas is measured with a pressure transducer either
once a week or at the end of test. In addition, the DIC (dis-
solved inorganic carbon) content of the digester liquid is de-
termined at the end of the test. From the cumulative net gas
production (net gas = gas in the test vessels minus gas in
controls) it is possible, by application of the gas laws, to cal-
culate the amount of test compound-derived organic carbon
transformed to gaseous one-carbon products (CH4, CO2). In
general, the incubation is finished when the cumulative net
gas production curve shows a plateau (i. e. when the gas pro-
duction rate in controls and test vessels is virtually compar-
able. The net DIC formation is obtained as the difference of
DIC concentrations between the test and control vessels.
The extent of anaerobic ultimate degradation is calculated
by comparison of the amount of carbon equivalent to net
gas and DIC production with the initially added organic car-
bon content of the test chemical.
Tenside Surf. Det. 44 (2007) 6
J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review
Figure 4 Principle and design of ECETOC Test/ISO 11734
Technical aspects
The method is applicable to water soluble and poorly soluble
substances provided the test concentration (20 – 100 mg C/l)
is not inhibitory to the test inoculum. As with the previous
test, inhibition can be easily recognised by the observation of
a negative net gas production, i. e. a gas production which is
higher in the control vessels than in the vessels containing
the test substance.
The organic carbon content of the test substance must be
known; additional substance information (chemical compo-
sition, theoretical CO2/CH4-ratio according to the Buswell
equation) is not necessary. This is a considerable advantage
over anaerobic screening methods only based on gas volume
measurement (cf. 6.1.1) since neither correction factors for
CO2 solubility nor assumptions of the theoretical ratios of
CO2 and CH4 formation are necessary. Thus, the ECETOC/
ISO 11 734 method is recommended when more accurate
values are needed for chemicals having a known empirical
formula, or if a value is required for a test material of un-
known composition but its percentage carbon is known.
(Painter 1994).
The ECETOC test and the standard tests based upon
have been adopted by many laboratories in Europe for sev-
eral years so that its applicability and practical usefulness
can be considered as broadly approved. Information about
the reproducibility of the test can be obtained from the de-
gradation data reporting on, respectively, the results of inde-
pendent tests of the same substance and the mean degrada-
tion value including its standard deviation when multiple
parallel assays are made (Pagga and Beimborn, 1993). Ac-
cording to these data it can be concluded that the ECETOC/
ISO 11 734 test provides reproducible results showing a var-
iation which is typical of biological test systems.
The ECETOC/ISO 11 734 test is, like other biodegradation
screening tests, a relatively simple test system not requiring
highly specialised technical staff, but nevertheless it does re-
quire sound expertise. Thus, it is strongly recommended
that inexperienced staff is trained in the use of the test by
testing well investigated model substances and comparing
the obtained results with published data.
Test modifications
A modified ECETOC test method was used by Madsen et al.
(1995) to determine anaerobic biodegradation potential in
digested sludge, a freshwater swamp and marine sediment.
Tenside Surf. Det. 44 (2007) 6
In the mineral medium FeCl2 was replaced by a trace ele-
ments mixture. The test medium was inoculated with 10 –
20 % fresh or washed domestic digester sludge (1.5 or
0.15 g of dry solids/l), with 5 % of the freshwater swamp
(24 – 88 g of organic carbon/kg), or with 10 % of the marine
sediment (9 g of organic carbon/kg), respectively. The gas
production in the headspace was measured by a pressure
transducer while CH4 was determined at the termination of
the incubation period by gas chromatography. Dissolved in-
organic carbon was quantified at the end of the test by acid-
ification of the liquid and subsequent gas pressure measure-
ment.
The authors suggest that the digester sludge inoculum
concentration be reduced to 0.15 g of suspended solids/l in
order to eliminate the sludge washing step usually required
in order to reduce the amount of inorganic carbon when a
higher concentrated inoculum is used. This however in-
creases the test concentration to biomass ratio unless the
test concentration is lowered accordingly.
6.1.3 Predictive value of anaerobic biodegradation screening test data
As already mentioned a common feature of biodegradation
screening tests is that they do not reproduce realistic envir-
onmental conditions and are more stringent than the tests
simulating the real world. Therefore, a poor degradation re-
sult is not necessarily a proof of recalcitrance in the real en-
vironment. Examples of anaerobic biodegradation results
supporting this argumentation have been published (Birch
et al. 1989, Steber 1991, Steber and Birch 1995a). On the
other hand, a positive result in an anaerobic screening test
can be considered as highly predictive for extensive biode-
gradation in anaerobic environments.
Although the production of CH4 and CO2 is the evalua-
tion criterion in the anaerobic screening tests, positive re-
sults in such tests may have relevance beyond methanogenic
systems. The major part of the anaerobic biodegradation
route (hydrolysis, fermentation, and acetogenesis) is com-
mon to all anaerobic pathways (cf. Fig. 2). These only differ
basically in terms of the final stage of the process, i. e. by the
different utilisation of the final electron acceptors like car-
bon dioxide for methane production, sulfate for sulphide
formation and nitrate for the production of ammonia and
molecular nitrogen, respectively. Thus, it can be expected
that a chemical being degraded in the discussed anaerobic
screening tests will also undergo biodegradation in those en-
vironmental conditions where the final degradation is
brought about by denitrifying or sulfate-reducing bacteria
(Steber et al. 1995b).
An additional aspect of the environmental relevance of
anaerobic screening test results also has to be kept in view.
The discussed tests determine the ultimate biodegradation
of a chemical by measurement of the production of the final
gaseous products, i. e. methane and carbon dioxide. There-
fore, it has to be acknowledged that the bacterial transforma-
tion of the parent chemical (primary biodegradation) by
anaerobic bacteria is not reflected by these data. Thus, even
a poor degradation result in such a screening test does not
necessarily indicate for anaerobic persistence of the parent
compound. In other words, if the ultimate degradation of a
substance under these test conditions is significant (‡ 20 %
gas production) it can be concluded that the primary biode-
gradation of this chemical is virtually complete as at least
20 % of the carbon atoms of each molecule have been miner-
alised. Obviously, this assumption is only valid for pure
compounds.
321
J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review

Figure 5 The anaerobic digester simulation

test

6.2 Simulation tests
6.2.1 Introduction to anaerobic simulation tests
The motivation to move beyond the screening level to higher
testing tiers can be:
y The need for increased environmental realism – includ-
ing obtaining realistic kinetic information for different
anaerobic environments, inocula, reactor types or test
conditions,
y To avoid inhibition of the anaerobic microorganisms by
the test material by working at a lower and more realistic
test material to biomass ratio,
y To increase the chances of acclimation by exposing a
broader range or organisms
y To improve the signal/noise ratio of the test,
y To study the CO2 /CH4 ratio
y To study the kinetics of the process.
The following test systems will be discussed:
14
1) C-Anaerobic Digester Simulation Test (batch of fed-batch
system)
2) Anaerobic Transformation test in Aquatic Sediment Systems
(OECD Guideline 308)
3) Continuously operated anaerobic reactors
4) Biological nutrient removal Simulation Tests and other systems
of this method is that the headspace of the anaerobic vessel
is continuously purged with N2, which is passed through a
series of base traps, to capture first the 14CO2 evolved. The
effluent gas is mixed subsequently with oxygen and passed
through a combustion tube (CuO at 800 – 900 8C) to convert
the 14CH4 into 14CO2. The latter is trapped in a second ser-
ies of base traps (see Figure 5). The test can be run in a
batch, or fed-batch mode (with addition of fresh sludge/test
material at regular intervals). The technical details of this
system are provided in Steber and Wierich (1987) and Nuck
and Federle (1996).
The anaerobic digester system, if well constructed and
operated, provides excellent recovery of the radioactive
gases, and therefore good mass balances. Given the simula-
tion of in-situ conditions and the realistic ratio of test sub-
stance: biomass, the system can generate relevant kinetic
data (Figure 6).
6.2.2.2 Anaerobic transformation test in aquatic sediment systems
(OECD guideline 308)
This guideline describes a laboratory test method to assess
aerobic and anaerobic transformation of organic chemicals
Alkyl ethoxy sulfate (AES) – % mineralized in anaerobic sludge
(Nuck & Federle, 1996)

Only the Aquatic Sediment Test (OECD308) has been ac-

cepted for international standardisation. Some of the other
systems are under evaluation as potential standard methods.

6.2.2 Test systems

14
6.2.2.1 C-Anaerobic digester simulation test (see Figure 5)

This test system also known as Anaerobic Mineralization

Test, assesses the mineralization of 14C-radiolabelled test
chemicals to CO2 and CH4 under anaerobic (methanogenic)
conditions. (Steber & Wierich, 1987; Gledhill, 1995; Nuck &
Federle, 1996). The inoculum is usually obtained from an
active digester but in principle the system can also be used
to simulate other anaerobic compartments, such as septic
tanks or sediments. The system follows the formation of Figure 6 Example of the anaerobic mineralization of A24E3S (ethoxylate la-
CO2 and CH4 separately over time. The underlying principle belled) to CO2 and CH4. Results are presented as % of the initial counts added

322 Tenside Surf. Det. 44 (2007) 6

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review
in aquatic sediment systems. An OECD Workshop on Soil/
Sediment Selection, held in 1995 agreed, in particular, on
the number and type of sediments for use in this test.
The surface layer of sediment can be either aerobic or
anaerobic, whereas the deeper sediment is usually anaero-
bic. The aerobic test simulates an aerobic water column over
an aerobic sediment layer that is underlain with an anaero-
bic gradient. The anaerobic test simulates a completely
anaerobic water-sediment system.
Principle of the test:
The method described in this guideline employs an aerobic
and an anaerobic aquatic sediment system which allows:
(i) the measurement of the rate of transformation of the
test substance in a water-sediment system,
(ii) the measurement of the rate of mineralization of the
test substance and/or its transformation products (when
14
C-labelled test substance is used),
(iii) the identification and quantification of transformation
products in water and sediment phases including mass
balance (when labelled test substance is used)
(iv) The measurement of the distribution of the test sub-
stance and its transformation products between the two
phases during a period of incubation in the dark (to
avoid, for example, algal blooms) at constant tempera-
ture.
The study is typically conducted in a biometer appara-
tus, or in gas flow-through systems. Half-lives, DT50, DT75
and DT90 values are determined where the data warrant,
but should not be extrapolated far past the experimental
period.
To the best of our knowledge and at the time of writing
there was no OECD 308 study available on surfactants, but
the system potentially lends itself to anaerobic sediment de-
gradation studies for this class of chemicals.
6.2.2.3 Continuously operated anaerobic reactors
Several types of semi-continuously and continuously oper-
ated anaerobic reactors have been used in the literature to
obtain information on the anaerobic degradability of surfac-
tants.
Continuously operated fixed bed systems
Continuously operated fixed bed systems have been pro-
posed by e. g. Bouwer and McCarthy (1983), Wagener and
Schink (1987) and Baumann and Mueller (1997) to study
the anaerobic degradation of soluble chemicals or waste-
waters. The reactors were filled with sinter-glass rings or
any other suitable support material. Inoculation was with di-
gester sludge and the system is maintained on a synthetic
medium. In the Wagener and Schink approach, the total
biogas volume was monitored, while in the EMPA system
(Baumann and Mueller, 1997) the biogas was collected in a
glass tube containing sodium hydroxide to absorb CO2. The
anaerobic degradability of a test substance can be calculated
from the maximum theoretical volume of biogas/CH4 and
the amount of biogas/CH4 actually produced.
The continuous anaerobic fixed bed system can be used
to perform simple mass balance studies, or according to the
authors, to gain information on hydrophilic and hydropho-
bic metabolites (provided adequate analytical methods are
available). A typical test concentration for a non-labelled
(non-toxic) chemical is in the order of 250 – 500 mg/l as
COD.
Tenside Surf. Det. 44 (2007) 6
Continuous stirred tank systems
Metzner & Lemmer (1997) illustrate a dynamic semi-conti-
nous system loaded intermittent with sludge from munici-
pal sewage digester. Due to varying sludge qualities of dif-
ferent origin, semi-continous systems are difficult to
stadardize. Also high background levels of organics, e. g. sur-
factants cannot be avoided. For this reason Metzner (2001)
suggested to use synthetic sludge in a thermostatically con-
trolled continous system. The possibility of automatization
leads to a lower susceptibility to faults and the advantage of
better standardization. Austermann-Haun et al. (2004) re-
port on the use of a thermostatically controlled continuously
stirred reactor system where the amount of biogass formed
is measured. No specific results for surfactants were re-
ported. This system can be run continously and/or semi-
continously as a degradation and inhibition test.
Metzner (1998) describes the absence of degradation and
the inhibition at higher concentrations of LAS in an anaero-
bic, semi-continously fed stirred tank reactor.
Between 2000 and 2003, three separate papers were pub-
lished by a group in Denmark describing the disappearance
of LAS in laboratory continuous stirred tank reactors (Angel-
idaki et al., 2000; Hagensen et al., 2002; Mogensen et al.,
2003). The experiments consisted of 3.5 L reactors semi-con-
tinuously fed with sludge solids spiked with LAS and moni-
toring by HPLC the level of LAS in the effluent over time.
The reactors had hydraulic retention times of 15 days and
were operated at 37 8C. They aimed to simulate a single
stage anaerobic digestor common in treatment plants. Only
a limited removal of LAS was observed in this reactor sys-
tem. The exact mechanism of removal (degradation and/or
sorption) was not fully clarified.
Upflow anaerobic sludge blanket systems
The same research group from Denmark published two se-
parate papers on the biodegradation of LAS in UASB reac-
tors (Mogensen & Ahring, 2002; Mogensen et al. 2003).
UASB reactors are commonly used to treat industrial waste-
waters and less for domestic wastewater. They contain
granulated sludge held in suspension by the upflowing
water. The authors chose this type of reactor to maximize
the bioavailability of LAS. Removal was determined in the
sterile and bioactive reactors. The effluents levels from the
bioactive unit levelled off at 6 mg/l, while those from the
sterile unit continued to increase up to 9 mg/l at 312 hours,
which suggests some degradation of LAS. Analyses of the
effluent from the bioactive unit revealed a transient 24 % in-
crease in volatile fatty acid production relative to the sterile
control upon addition of LAS to the feed. In addition, total
ion chromatograms revealed several peaks in the effluent of
the bioactive unit not found in the effluent of the sterile con-
trol or a unit fed only minimal salts medium without LAS.
One of the papers identifies the presence of benzene sulfo-
nate (Mogensen & Ahring 2002) and another identifies the
presence of benzaldehyde (Mogensen et al. 2003). Neither
paper mentions both materials together. There is no indica-
tion on the level of either metabolite that can provide insight
into the stoichiometry of the reactions.
A paper by a group in Spain (Sanz et al, 2003) also refers
to UASB reactors, operated at 30 C. One reactor was fed or-
ganic nutrients plus 4 – 5 mg LAS/l and the other was fed
only LAS in mineral medium. Based upon the total mass
balance, 64 % of the LAS was lost from the first reactor and
85 % from the LAS-only reactor. There was no sterile control,
nor data on gas production and metabolites in the effluent.
323
J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review

6.2.2.4 Biological nutrient removal STP simulation tests: Since these would be considered as non-routine tests,
Behr and CAS-UCT units they will not be covered further.

Wastewater treatment systems with biological nutrient re-
moval are becoming quite common across Europe (http://
epp.eurostat.ec.europa.eu). The Behr- and CAS-UCT test
systems are modifications of the OECD303A method and al-
low to simulate the behaviour of chemicals (e. g. surfactants)
in laboratory-scale sewage treatment plants with anoxic
(Behr unit) and anaerobic plus low dissolved O2 zones
(CAS-UCT system) (Schowanek et al., 1996). These are typi-
cal configurations for single-sludge wastewater treatment
plants with biological N/P removal. While these are not en-
tirely anaerobic biodegradation test systems, they have been
included in this chapter for information purposes. The
Behr- and CAS-UCT laboratory units can also be constructed
in a way which allows 14C-mass balance studies.
6.2.2.5 Others systems
Various other anaerobic reactor designs (e. g. septic tank,
anaerobic contact process, aquifer column,) can also be oper-
ated on a laboratory-scale to study the behaviour of chemi-
cals in these specific systems or environments (e. g. Kuhn
et al., 1988; Heijnen et al., 1991).
7 Interpretation of Available Data on Anaerobic
Biodegradation of Surfactants
This chapter gives a compilation of existing literature data
on anaerobic biodegradation of all surfactant classes.
Data on laboratory tests are shown in individual tables
together with a discussion of the results.
Monitoring data of surfactants in anaerobic compart-
ments reported in the literature do not necessarily reflect
the biodegradation behaviour of surfactants under strictly
anaerobic conditions. For example, temporary aerobic condi-
tions in an otherwise anaerobic environment may influence
the results towards a high degree of degradation. Therefore
such monitoring data (see Appendix) may not be used as a
proof of biodegradability/recalcitrance in anaerobic environ-
ments, but may form an additional support of the conclu-
sions drawn from the laboratory test results.
7.1 Anionics
7.1.1 Sulfonates
In tables: Separate lab studies from the field studies

Sur- Characteri- Test type Test conc. Inoculum Test Temp. Results Remarks References
fac- sation (dw) conc. Dura-
tant tion
type
Screen- Simulation in mg/l mg/l in g/l Days 8C
ing active car-
matter bon
LAS C10 7 C13 ECE- 50 1–5 49 35 0 Steber (1991)
commercial TOC
LAS ring-14C digester 10 20 27 37 0.3 14C LAS Steber (1991)
LAS ring-14C field system 0.5 WW pond 87 22 0 14C LAS Federle &
sed. Schwab (1992)
LAS C12 LAS ECE- 50 60 inhibition bio- Battersby &
TOC gas formation Wilson (1989)
LAS C8 LAS ECE- 50 60 inhibition bio- Battersby &
TOC gas formation Wilson (1989)
LAS C10 7 C13 field system septic tank 97 (a) 10 % 14C LAS Klein and
commercial Mehaughey
(1964)
LAS C10 7 C13 field system 10 99.7 (a) Larson et al.
commercial (1989)
LAS C12 LAS research 5 sediment 6 100 (k < 0.23 d-1) 14C LAS Heinze &
study slurry (b) Britton (1994)
LAS C10 7 C13 research 5 – 20 0.02 – 0.1 29 h 25 100 (k = 0.14 h-1) specific LAS Denger & Cook
commercial study (c) and S (1999)
LAS 2-Ph C10 Birch research 20 Anaerobic 49 Negligible both for 14C LAS en- Garcia M. T.
LAS, C12 et al. study sludge days LAS and for SPC hances biogas et al. (2005)
and C14 1989 Same results with production
Ph in position 5
LAS C12 LAS a-Semi-con- 100 Anaerobic 15 37 20 % biodeg. Mogensen A. S.
tinuous stir- sludge days et al. (2003)
red tak reac-
tor CSTR 12
b-UASB 10 – 20 hrs 55 37 % biodeg. UASB
LAS C10 7 C13 UASB up 1 UASB ther- 12 40 % biodegrada- Benzene sul- Mogensen A. S.
commercial flow anaero- mophilic hrs tion fonic acid and and Ahring B. K.
bic sludge granular benzaldehyde (2002)
bed sludge were found

324 Tenside Surf. Det. 44 (2007) 6

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review

Sur- Characteri- Test type Test conc. Inoculum Test Temp. Results Remarks References
fac- sation (dw) conc. Dura-
tant tion
type
Screen- Simulation in mg/l mg/l in g/l Days 8C
ing active car-
matter bon
LAS LAS C12 CSTR 20 Digested 8 hrs 37 40 – 80 % removal Specific LAS Angelidaki, I.
LAS 14C12 UASB sludge and 50 % absorbed on analyses et al. (2003)
55 sludge and (2004)
5.6 % degradation
by 14CO2
LAS LAS C12 CSTR Anaerobic 14 – 25 % primary Specific LAS Haggensen, F.
sludge biodegradation analyses et al. (2002)
LAS C10 7 C13 ECE- Anaerobic Poor primary bio- Garcia, et al.
commercial TOC digester deg. for single LAS (2004)
isomers
LAS LAS C10 – Laboratory Marine se- LAS absorption on Equilibrium: Saes M. et al.
13 study diments marine sedum. and 12 hrs for LAS (2003)
organisms and 18 hrs for
SPC
LAS LAS C10 – ECE- Anaerobic 4 – 22 % primary 4 – 26 % inhi- Garcia et al.
13 TOC sludge bition of biogas (2006)
production
LAS LAS C10 – ECE- Laboratory Anaerobic LAS at conc. < 5 – Garcia, Campos
13 TOC study on LAS sludge 10 g/kg dw in- et al. (2006)
effect on the crease biogas pro-
anaerobic di- duction; inhibition
gestion of at higher concen-
sewage trations
sludge 14.5 m/l estimated
LAS toxicity on
anaerobic micro-
orgs
LAS LAS C10 – Laboratory 10 – 50 Marine se- 165 79 degradation Proof of the Pablo A. Lara-
13 study diments within 165 days via primary biode- Martín et al.
commercial the generation of gradation of (2007)
sulfophenyl car- LAS under
boxylic aids anaerobic con-
dit.
MES 2-sulpho digester 10 20 28 35 0.6 14C MES Steber &
(14C) palmi- Wierich (1989)
tic Me ester
SAS C14 7 C17 ECE- 17 0 Bruce et al.
TOC (1966)

a) Oxygen – limited conditions

b) Mineralization kinetic study in different oxygen-limited conditions
c) Desulfonation kinetic study using an enriched and isolated strain. Primary biodegradation

Conclusions on the anaerobic biodegradability of sulfonates
1) Laboratory data
Sulfonates are not degraded significantly under the anaero-
bic conditions of the laboratory test methods or in anaerobic
digesters of sewage sludge (Steber, 1991, Mogensen 2002,
Mogensen 2003, Angelidaky 2004, Garcia 2004 and Garcia
2005).
In the real environment and also in field system tests,
however, oxygen-limited conditions may be more common
than rigorously anaerobic conditions. In such conditions
sulfonates mineralize even if the rate is not as rapid as that
observed under aerobic conditions. Once sulfonate biodegra-
dation has been initiated in aerobic or oxygen-limited condi-
tions, the intermediates can continue to biodegrade anaero-
bically. This is the reason why in some simulation tests
these chemicals can show mineralization results, if some
oxygen diffusion had occurred or if limited-oxygen condi-
tions had been created (Larson, 1989; Heinze and Britton,
1994). On the other hand, it must be also considered that
only 0.2 mg/l of dissolved oxygen can support biodegrada-
tion if O2 recharge to the aquifers is not limited (Salanitro
1997).
It was observed that LAS can inhibit biogas formation in
anaerobic biodegradation batch tests if present at high con-
centrations (Battersby and Wilson, 1989). It has also been
found however that LAS can increase the biogas production
when present at lower concentrations. In fact, Garcia and
Campos 2006 determined that the inhibition starts at LAS
concentrations of about 5 – 10 g/kg dw of LAS as Na salts
while, at lower concentrations the biogas production is in-
creased. Therefore, at the usual LAS concentration range in
sewage sludge, no adverse effects on the anaerobic digesters
of a wastewater treatment plant (WWTP) can be expected. In
addition, in real anaerobic environmental compartments
(e. g. STP anaerobic digester), however, sulfonates have
shown not to inhibit biogas formation even at high concen-
tration (> 30 g/kg dw) because they are present as Ca salts
which are poorly soluble and less bio-available (see 8.2).

Tenside Surf. Det. 44 (2007) 6 325

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review
It has also been demonstrated that not all desulfonation
reactions require oxygen. Desulfonation of alkylsulfonates
as well as of LAS has been reported to occur by anaerobic
bacteria in the laboratory (Denger and Cook, 1999). There
is recent evidence that anaerobic desulfonation can take
place. Desulfonation with assimilation of the sulphur moi-
ety by strictly anaerobic bacteria (Chien et al., 1995; Denger
et al., 1996; Denger and Cook, 1997) was followed by the re-
duction of the sulfonate group as a source of electrons and
carbon under anaerobic nitrate-respiring conditions (Lie et
al., 1996; Laue et al., 1997; Denger et al., 1997). There is no
evidence however that such mechanisms would also occur at
significant rates under real-world conditions.
Two breaking through publications of Pablo A. Lara-Mar-
tn et al. (2007) proved for the first time the degradation of
linear alkylbenzene sulfonates (LAS) under anaerobic condi-
tions through the generation of sulfophenyl carboxylic acids
(SPCs), together with the presence of metabolites and the
identification of microorganisms involved in this process.
Laboratory experiments performed with anoxic marine sedi-
ments spiked with 10 – 50 ppm of LAS demonstrated that its
degradation reached 79 % in 165 days via the generation of
SPCs. Almost all of the added LAS (> 99 %) was found to be
attached to the sediment while the less hydrophobic SPCs
were predominant in solution, as their concentration in-
creased progressively up to 3 ppm during the full course of
the experiment. The average half-life for LAS was estimated
to be 90 days. The researchers found that the degradation is
reduced when the amount of LAS in sediment exceeds
20 ppm, as this concentration could be considered toxic to
certain marine microorganisms. However, these LAS values
are unlikely to occur in aquatic ecosystems except for specif-
ic locations near untreated wastewater discharges. Sulfate-
reducing and methanogenic activities proved to be intense
during the experiment. Several sulfate-reducing bacteria
and firmicutes/clostridia were identified as possible candi-
dates for effecting this degradation.
2) Monitoring data (see Appendix)
Sludge
LAS concentrations in sludge have been reported in several
situations ranging from < 100 mg/kg to a maximum of
30,000 mg/kg (Berna, 1989) depending on emission vol-
ume, STP operating conditions and water hardness. Aerobic
stabilised sludge always has a LAS content typically lower
than 500 mg/kg dry matter, whereas anaerobically digested
sludge has a LAS concentration in the range 1,000 –
30,000 mg/kg, with a mean value around 5,000 mg/kg.
(Waters, 1995 and McAvoy et al., 1993, and Schowanek et
al. in press). The latter authors present a distribution of
LAS concentrations found in anaerobic sludges, based on
data reported in the scientific literature and (industry) re-
ports, covering different European countries, and the time
period 1988 – 2006. Results are overall quite consistent be-
tween different studies, and between the EU and the US.
A few studies have attempted to make a LAS mass bal-
ance over full scale anaerobic digesters. A low degree of re-
moval is measured, ranging from 0 – 35 % (Berna, 1989 and
Giger, 1987). These data fit broadly with the results from la-
boratory screening or digester tests, and biochemical in-
sight, which indicate that LAS, as well as any other sulfonate
surfactants, does not degrade under strictly anaerobic condi-
tions. It is not entirely clear to which process (as) the small
degree of removal observed in field systems can be adscribed
(binding, humification, co-metabolism, and anaerobic desul-
fonation). LAS concentrations on a wet sludge basis actually
326
increase versus fresh aerobic sludge in an anaerobic digester
due to the dewatering and digestion of solids.
We are aware of only another study on anaerobic degra-
dation of sulfonates in full scale systems, not related to LAS
but instead to SAS (Field, 1992). The SAS monitoring in
anaerobic sludge ranges from 270 to 800 mg/kg. Like LAS,
there is no evidence for a significant anaerobic degradation
of SAS.
Sediments
LAS has been found in river sediments. The levels found
just below an STP outfall range from zero to a maximum
value of 174 mg/kg, (Rapaport, 1990). Downstream the
same STP outfall the LAS concentrations in sediments drop
to 5 – 11 mg/kg (Rapaport, 1990). In Po river sediments, con-
centrations from 0.4 to 4.7 mg LAS/kg were detected (Cavalli
2000). No concentration effect was observed along 500 km of
the river, indicating that there is no LAS accumulation de-
spite the high load from this big catchment.
LAS has also been detected in marine sediments at con-
centrations ranging from not detectable to few ppm, (Del
Valls 2002, Folke 2003, Bester 2001, Leon 2001, Lara-Matin
2006). Higher concentrations were found only in polluted lo-
cations like 10 mg/kg in one site used for direct discharge
(Lara-Martin 2005) or 20 mg/kg in a Danish port (Folke
2003).
No mass balance studies are known to us. Several inter-
esting studies have looked at the levels of LAS and other sur-
factants in cores or river/lake sediments as accumulated
over time. Residual LAS levels can be measured, and in the
trends over time (i. e. depth) clear records can be seen re-
flecting the switch from branched alkyl sulfonates to LAS
(Schçberl, 1996; Reiser, 1995).
Soil
LAS and SPC were determined in sludge amended soils of
10 field sites in the North of Spain. The sludge application
was generally at the rate between 8 and 20 tonnes per hec-
tare and the sites were treated at least once over the period
1997 – 2002. The concentrations were in the range 0.12 –
2.8 mg /kg and 0.004 – 0.2 mg/kg. respectively for LAS and
SPC. The highest concentration was found in the field that
had received sludge 10 days before the sampling (Eichhorn
2005). The authors concluded that, applying the estimated
PNEC of 4.6 mg /kg (Jensen 2001) LAS do not pose a signif-
icant risk to fauna, plants and essential functions of agricul-
tural soils.
In another Danish study that includes 8 locations with
different histories, including sludge amended soil, it was
concluded that it is not possible to note significant differ-
ences between undisturbed soils used for grazing for 50 –
100 years and soils being moderately sludge amended. All
LAS concentrations were in the range 0.5 – 1.0 mg/kg. Only
in an heavy sludge amended location was found 10 – 20 mg
LAS/kg but it was not representative because the amend-
ment of this location was carried out at a high rate/17.5 t
dw/ha/y for 25 years/, therefore not respecting the currently
adopted EU regulation (Carlsen 2002).
These low concentrations of LAS in soil can be explained
by the fact that LAS can be used by bacteria as a source of
energy and carbon (Elsgaard 2003), and rapidly degrades in
soil with a half life of ca. 10 days (primary degradation) and
30 days (mineralisation) (Schowanek et al., in press).
Several risk assessments of LAS in the terrestrial envir-
onment have been published over time and have introduced
subsequent refinements (Mieure et al. 1990; De Wolf and
Feijtel 1998; Jensen 1999; Jensen et al. 2001; HERA 2004).
Tenside Surf. Det. 44 (2007) 6
J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review

Recently, the PNECsoil has been revised upwards from 4.6
to 35 mg/kg (Jensen et al., 2007). Based on this work, Scho-
wanek et al. (in press) have performed a deterministic and
probabilistic risk assessment for LAS in anaerobic sewage
sludge. The conclusion, backed up by relevant field evi-
dence, is that LAS in anaerobic sludge does not represent a
significant ecological risk. From this perspective there is no
scientific basis for specific regulatory limits for LAS in
sludge as present in earlier EU draft legislation, nor the
need to consider specific sludge management options due
to the presence of LAS.
7.1.2 Sulfates

Surfactant Characterisa- Test type Test concentration Inocu- Test Temp. Results Remarks References
type tion lum dura-
conc. tion
Screening Simula- in mg/l in mg/l in g/l in 8C %
tion active carbon days
matter
14
Alcohol C12 Digester 10 5 20 28 35 90 C-labelled Steber et al.
Sulfates (31 % CO2 (1988)
+ 59 % CH4)
C12 – 13 ECETOC 30 1–5 42 32.6 – 70 Biogas + IC Rehman et al.
linear 36.8 (2005)

C12 – 13 mid- 40
chain
branched

C12 – 13 25
mainly
branched in
ß-position
C12 – 14 Mod. 96 50 1 40 – 35 77 – 84 CH4 Salanitro & Diaz
Shelton & 50 (1995)
Tiedje 98 – 99 MBAS

C12 – 18 ISO 100 1.5 84 35 59 Biogas + IC Fraunhofer

14853 99 MBAS (2003)
14
C14 Digester 1 0.5 26 15 35 80 C-labelled Nuck & Federle
(CO2 + CH4) (1996)
C14 – 15 Mod. 93 50 1 40 – 35 65 – 78 CH4 Salanitro & Diazl
Shelton & 50 (1995)
Tiedje 97 MBAS

C14 – 15 ECETOC 30 1–5 42 32.6 – 60 Biogas + IC Rehman et al.

80 % linear 36.8 (2005)
C18 ECETOC 50 29 3 56 35 88 Biogas + IC ECETOC (1988)
14
C18 Digester 10 6 20 28 35 94 C-labelled Steber et al.
(43 % CO2 (1988)
+ 51 % CH4)
Alcohol C12, xEO Mod. 40 – 100 20 – 0.06 – 55 – 35 14 – 41 Biogas (in- Madsen & Ras-
Ethoxy ECETOC 50 0.12 56 cludes IC after mussen (1994)
Sulfate acidification)
C12 – 14, ECETOC 50 1–5 41 35 75 Biogas + IC Steber (1991)
2 EO
C12 – 14, ISO 100 1.5 84 35 o Biogas + IC Fraunhofer
2 EO 14853 73 MBAS (2003)
50 1.5 119 35 60 Biogas + IC
14
C14 – 15, Digester 1 – 10 0.5 – 5 26 17 35 88 C-labelled Nuck & Federle
2 EO (CO2 + CH4) (1996)
C14 – 18, Septic 26 – 52 14 – 29 No 240 No 72 – 81 MBAS Painter (1992)
3 EO tank data data

Conclusions on the anaerobic biodegradability of sulfates
Alcohol Sulfates
1) Laboratory data
Linear primary alcohol sulfates with alkyl chain lengths typi-
cally used in detergents (C12 – 18) are anaerobically biode-
gradable, with a conversion to CH4 and CO2 (biogas) of
80 – 94 % in tests simulating anaerobic digesters. They are
also extensively mineralised (59 – 88 %) in screening tests,
although as with other anionic surfactants, the unrealisti-
cally high surfactant to biomass ratio means that low biode-
gradation or inhibition of biogas production are sometimes
observed. Branching of the alkyl chain results in a reduced
extent of ultimate anaerobic degradation under screening
test conditions. The primary biodegradation of alcohol sul-
fates under anaerobic conditions is very high exhibiting a
MBAS reduction of ‡ 97 % in screening tests.

Tenside Surf. Det. 44 (2007) 6 327

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review

2) Monitoring data formation may imply a high percentage of primary biode-

Sludge
No monitoring data are available.
Sediments
In a monitoring study at the Bay of Cdiz, Spain, (Lara Mar-
tn et al., 2005) AS even alkyl chain homologues from C12 to
C18 AS were found in very low concentrations (< 100 ppb),
while no levels of AS with odd homologues were detected in
most of the sampling stations due to their low usage in com-
parison to other high volume surfactants like LAS or AES.
Soil
No monitoring data are available.
Alcohol Ethoxysulfates
1) Laboratory Data
Compared to alcohol sulfates less experimental data are
available on the anaerobic biodegradability of ethoxysulfates
having alkyl and EO chain lengths typical of the use in deter-
gents (C12 – 18, £ 4EO). However, the existing information
indicates that they are also extensively biodegraded (‡ 60 %
biogas formation) under anaerobic conditions. Low biode-
gradation has been reported in some cases, e. g. for AES,
but this can be attributed to the very high test substance to
biomass ratio used in the study quoted. Similarly, the re-
ported data from ISO 14 853 screening tests show the depen-
dence of the results from the test concentration. In addition,
it becomes evident that even test results with poor biogas
gradation (MBAS reduction).
2) Monitoring Data
Sludge
No monitoring data are available.
Sediments
A research group from the University of Cdiz, Spain, inves-
tigated the distribution of AES homologues in sediments
taken from the coastal area of Spain (Gonzlez Mazo et al.,
2005).
Concentrations of AES in those marine sediments have
been found to be very low in the Bay of Cadiz (Spain) as well
as in other bays of the Spanish coast. Typical concentration
values of AES homologues in 1 cm depth were ranging be-
tween 150 and 400 ppb. The main trend along the sedimen-
tary column is towards a decrease in the concentration va-
lues of all these compounds with depth, with no detection
or a non possible quantification due to very low concentra-
tions in most cases at 17 cm and especially 47 cm. An addi-
tional monitoring study at the Bay of Cdiz was carried out
(Lara Martn et al., 2005) finding concentrations of AES of
125 ppb.
Soil
No monitoring data are available.
7.1.3 Fatty acids and soaps

Surfactant Characterisa- Test type Test concentr. Inoculum Test Temp. Re- Remarks References
type tion concentration dura- sults
tion
Screening Simula- in mg/l car- in g/l Days 8C %
tion active bon
matter
Fatty acid dodecyl (C 12) ECETOC 20 0.15 sludge 56 35 > 75 gas after Madsen et al.
acidific. (1995)
ECETOC 20 4.4 organic C/l 56 35 > 75 gas after Madsen et al.
(freshwater acidific. (1995)
swamp sedi-
ment)
ECETOC 20 0.9 organic C/l 96 35 > 75 gas after Madsen et al.
(marine sedi- acidific. (1995)
ment)
coconut ECETOC 18 0.06/0.12 55 35 40 – gas after Madsen &
(C 12 – 18) sludge 57 acidific. Rasmussen
(1994)
(u*-14C) static 10 20 sludge 28 35 96.5 56.6 % Steber &
palmitic (C 16) CH4 + 39.9 % Wierich
CO2 (1987)
Soap Na-laurate semicont. 200 30 sludge 20 (re- 35 95 CH4 meas- Mix-Spagl
(C 12) tention ured, CO2 (1990)
time) calc.
Ca-laurate static 1000 30 sludge 5–6 35 90 CH4 meas- Petzi (1989)
(C 12) ured, CO2
calc.
Na-palmitate ECETOC 70 1 – 5 sludge 28 35 94 gas + DIC Birch et al.
(C 16) (1989)
Na-palm semicont. 200 30 sludge 20 (re- 35 67 CH4 meas- Petzi (1989)
kernel tention ured, CO2
(C 8 – 18) time) calc.
Na-tallow semicont. 200 30 sludge 20 (re- 35 60 CH4 meas- Petzi (1989)
(C 16/18) tention ured, CO2
time) calc.

* uniformly marked

328 Tenside Surf. Det. 44 (2007) 6

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review

Surfactant Characterisa- Test type Test concentr. Inoculum Test Temp. Re- Remarks References
type tion concentration dura- sults
tion
Screening Simula- in mg/l car- in g/l Days 8C %
tion active bon
matter
Soap Na-stearate semicont. 200 30 sludge 20 (re- 35 51 CH4 meas- Mix-Spagl
(C 18) tention ured, CO2 (1990)
time) calc.
Ca-stearate static 1000 30 sludge 10 35 85 CH4 meas- Mix-Spagl
(C 18) ured, CO2 (1990)
calc.
Na-oleate semicont. 200 30 sludge 20 (re- 35 69 CH4 meas- Mix-Spagl
(C 18 unsat.) tention ured, CO2 (1990)
time) calc.
Na-behenate semicont. 200 30 sludge 20 (re- 35 14 CH4 meas- Mix-Spagl
(C 22) tention ured, CO2 (1990)
time) calc.
Ca-behenate static 1000 30 sludge 10 35 90 CH4 meas- Mix-Spagl
(C 22) ured, CO2 (1990)
calc.
Na-Laurate ECETOC static 100, 200, 54 35 94 – gas + DIC Varo et al.
(C 12) 400, 600 100 (2002)
Na-Laurate ECETOC static 1000 54 35 0 gas + DIC Varo et al.
(C 12) (2002)
Na-Stereate ECETOC static 100, 200, 54 100 gas + DIC Varo et al.
400, 600, (2002)
800

Conclusions on the anaerobic biodegradability of fatty acids and
soaps
1) Laboratory data
Fatty acids and their sodium/calcium salts (soaps) are well
biodegradable under anaerobic conditions. This was shown
in several laboratory studies using different test methods
(see Table). The data for dodecanoate and palmitate obtained
in the stringent ECETOC screening test showed very high
mineralization rates (> 75 % CO2 + CH4 formation) within
the test period of 4 – 8 weeks. The bacterial inocula used in
these investigations were digester sludge as well as anaero-
bic sediments from fresh water and marine environments.
The data for C12 – 18 fatty acids reported by Madsen and Ras-
mussen (1994) are lower (40 – 57 %) but this may indicate
slower biodegradation kinetics due to the unusually high
test substance/inoculum ratio used in the test. The positive
evaluation of the anaerobic biodegradability of fatty acids
and soaps was confirmed in a digester simulation study
using radio labelled palmitate and showing an almost quan-
titative ultimate degradation to carbon dioxide and methane
(Steber and Wierich 1987). Additional static and semi con-
tinuous digester simulation tests proved the extensive ulti-
mate biodegradation of Na- and Ca-salts of fatty acids with
an alkyl chain length of 8 – 22 carbons (Petzi 1989, Mix-
Spagl 1990). While the Ca-soaps (C12, C18, C22) exhibited
high gas formation rates (‡ 85 %) in the static system within
a 10-day test duration, the semi continuously run investiga-
tions with Na-soaps showed that the time needed for miner-
alization was increasing with the chain length and concomi-
tantly with the water solubility. It should be noted the all
these data refer to ultimate biodegradation based on the
measurement of gas production.
2) Monitoring data (see Appendix)
Sludge
Anaerobic biodegradation of soap has been also observed in
continuous laboratory scale equipment as well as on com-
mercial scale sewage treatment plant (D. Prats et al., 1999).
Average removal in both cases was in the 64 – 90 % range
being the lower values obtained for unsaturated homologues
regardless the residence time (25, 40 and 60 days). The fact
that no higher removals were observed can be attributed to
poor bioavailability of Ca–Mg soaps. Besides, fatty acids
might be potentially formed during the digestion process
from manifold chemicals of natural origin (e. g., lipids).
These results confirm previous mass balances of soap based
on a monitoring study of the concentrations in sludge of a
digester influent and effluent (Moreno et al. 1993).
Soil
The soap content was measured in sludge amended and non
amended soils from the southeast of Spain (Alicante). Soap
concentration in the untreated soil (blank) was 0.57 g/kg.
Once the soil was amended the soap initial concentration
was 1.47 g/kg. The concentration of soap 150 days after
amendment was below the value at the moment of the
amendment and can be considered therefore as totally bio-
degraded after this period. A steady state concentration of
soap might indicate the residual soap present as non bioa-
vailable form (D. Prats et al., 1999).
Sediments
There is a paucity of data about the presence of soap in ma-
rine sediments. The existing data evidence soap concentra-
tios in the 500 – 9000 mg/kg range (Folke et al., 2003; Lpez
et al., 2002). Although these studies cannot pinpoint the de-
tergent-origin of the detected soap, its concentration in rela-
tively uncontaminated sediments is from 15,000 up to
40,000 times higher than the LAS levels.

Tenside Surf. Det. 44 (2007) 6 329

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review

7.2 Nonionics

Surfac- Characterisation Test type Test Concentra- Inoculum Test Temp Results Remarks References
tant tion Concen- dura-
type tration tion
Screening Simu- in mg/l in mg/l days 8C
lation active carbon
matter
Alcohol C 9 – 11, 8 EO ECETOC 20 – 0.15 g dry 56 35 > 75 % Inoculum: Madsen et al.
Ethoxy- 50 matter/l digested sewage (1995)
lates sludge, fresh-
water swamp
C 9 – 11, 8 EO ECETOC 20 – 0.15 g dry 56 35 30 – 75 Inoculum: mar- Madsen et al.
50 matter/l ine sediment (1995)
C 9 – 11, 8 EO Mod. ISO 20 – 0.6 g dry 55 35 65 – 82 % Inoculum: Madsen & Ras-
11 734 50 matter/l biogas digester sludge mussen (1994)
(1995)
C 10 – 12, 7,5 EO screening 10 – 37 70 % biogas Inoculum, anoxic Wagener &
1000 freshwater Schink (1987)
sediment
C 12, 23 EO screening 10 – 37 80 % biogas Inoculum, anoxic Wagener &
1000 freshwater Schink (1987)
sediment
C 12, 23 EO simula- 1000 90 > 90 % CH4, Laboratory-scale Wagener &
tion acetate, anaerobic fixed- Schink (1987)
propionate bed reactor fed
with synthetic
wastewater
C 12, 9 EO 14C- 1,7 ug/ 87 22 13 – 40 % WW pond sedi- Federle &
screen g sedi- 14C; glu- ment Schwab (1992)
ment cose: 40 %
Isotridecanol, ECETOC, 20 2 – 3 g/l 110 35 0 – 30 % Siegfried et al.
(5, 10, 20) EO modified as solids ThGP (1996)
linear primary ECETOC, 20 0.15 g dry 35 % ThGP Inhibition during Madsen et al.
C 12-C15 7 EO modified matter/l the first 3 weeks (1996)
Linear C 12 – 14, ECETOC, 20 2 – 3 g/l 110 35 29 – 94 % Siegfried et al.
(5, 10, 20) EO modified as solids ThGP (1996)
mono br. C 14 – ECETOC, 20 2 – 3 g/l 89 35 0 – 23 % Siegfried et al.
15, (10,20) EO modified as solids ThGP (1996)
C 18, 7 EO 14C- 10 12 – 25 g 28 35 83 – 87 % Steber & Wierich
sim. dry mat- 14C as bio- (1987)
ter/l gas
Linear C 12-EOn laboratory 40 mg/l anaerobic 40 85.1 % Huber et al.
sludge (2000)
Alkyl- C 10 – 12 alkyl- screening 10 – 37 45 – 50 % Inoculum: anoxic Wagener &
phenol phenol, 9 EO 1000 CH4 freshwater Schink (1987)
Ethoxy- sediment
lates
Nonylphenol, ECETOC 50 1 – 5 mg 84 35 20.5 ± Steber (1991)
10 EO dry mat- 12.6 %
ter/l CO2 + CH4
Nonylphenol, ECETOC 50 1 g dry 40 – 35 32 – 43 % Salanitro & Diaz
9 EO matter/l 50 CH4 (1995)
Nonylphenol, simula- NP is metabolite Tschui & Brunner
0/1/2 EO tion of APEO, (1985)
accumulates
Glucose
Deriva-
tives
Gluco- Ethyl 6-O-de- Mod. ISO 20 56 35 59 – 65 % Inoculum: diges- Madsen & Ras-
side canoyl glucoside 11 734 biogas ter sludge mussen (1994)
(1995)
APG (branched) ECETOC, 30 – 20 0.15 g dry 22 % ThGP Primary domestic Madsen et al.
C8, DP = 1.6 modified 40 matter/l sludge (1996)
APG (linear) ECETOC, 30 – 20 0.15 g dry 72 % ThGP Primary domestic Madsen et al.
C 12 – 14, modified 40 matter/l sludge (1996)
DP = 1.4

330 Tenside Surf. Det. 44 (2007) 6

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review

Surfac- Characterisation Test type Test Concentra- Inoculum Test Temp Results Remarks References
tant tion Concen- dura-
type tration tion
Screening Simu- in mg/l in mg/l days 8C
lation active carbon
matter
Gluco- C 12 Ethylgluco- ECETOC, 30 – 20 0.15 g dry 82 % ThGP Primary domestic Madsen et al.
side side monoester modified 40 matter/l sludge (1996)
EGE
C 12 6-O-Ethyl- ECETOC 20 – 0.15 g dry 56 35 > 75 % Inoculum: Madsen et al.
glucoside 50 matter/l digested sewage (1995)
monoester sludge, fresh-
water swamp
C 12-C14 APG ECETOC 20 – 0.15 g dry 56 35 > 75 % Inoculum: Madsen et al.
50 matter/l digested sewage (1995)
sludge, fresh-
water swamp
C 10 6-O-Ethyl- ECETOC 20 – 0.15 g dry 55 35 > 75 % Inoculum: Madsen et al.
glucoside 50 matter/l digested sewage (1995)
monoester sludge, marine
sediment, fresh-
water swamp
C 12 – 14-APG ECETOC 100 3 g dry 56 35 84 ± 15 % Steber et al.
matter/l (CO2) + c. (1995)
(CH4)
C 8 – 10-APG ECETOC 100 3 g dry 56 35 95 ± 22 % Steber et al.
matter/l (CO2) + c. (1995)
(CH4)
Glu- C 12 Glucose 14C- 1 35 86 ± 0,3 % Federle and Nuck
cose Amide sim. (CO2 + (1997)
Amide CH4)
Amine Dimethyl Dode- 14C 1 ~20 7 35 > 80 % Vandepitte and
Oxides cyl Amine Oxide Diges- Debaere (1997)
ter
Alkyl- Coco monoetha- ISO 11734 20 1–3 56 35 81 % of the- Madsen (2001)
ethanol- nolamide oretical gas
amides production

DP = degree of polymerization
APG = alkylpolyglucoside
THGP = theoretical gas production

Conclusions on the anaerobic biodegradation of nonionic In a number of EU countries concentrations of alcohol

surfactants
Alcohol Ethoxylates
1) Laboratory data
Alcohol ethoxylates (linear C9–C18-alcohols, 5 – 23 EO) are
well biodegradable in anaerobic screening tests. Degradation
of usually > 70 % (biogas) has been reported in digested sew-
age sludge and freshwater sediment.
Degradation > 80 % (biogas formation and 14C) in diges-
ter simulation tests have been reported in the case of lauryl-
alcohol ethoxylates and stearylalcohol ethoxylates. In a study
on the biodegradation mechanisms of linear alcohol ethoxy-
lates under anaerobic condition it was concluded that the
first step of anaerobic microbial attack is the cleavage of the
terminal EO unit, releasing acetaldehyde stepwise, and
shortening the ethoxy chain until the lipophilic moiety is
reached (Huber et al., 2000).
2) Monitoring data
Sludge
Actual data on the concentration of alcohol ethoxylates in di-
gester sludge support the conclusion that alcohol ethoxylates
are well biodegradable in full scale digestors (Klotz, 1998).
ethoxylates were monitored in inlet and outlet sludges
streams. The removal of alcohol ethoxylates by anaerobic di-
gestion was on average 82 % (range 61 – 93 %). The applied
analytical method determined alcohol ethoxylate components
with C12–C18 alkyl chain lengths and an ethoxylate chain
length of 4 – 20 EO in sludge samples (Matthijs et al., 2004).
Concentrations of alcohol ethoxylates were monitored in
inlet and outlet sludges streams of a number of STP in dif-
ferent EU countries. The removal of alcohol ethoxylates by
anaerobic digestion was on average 82 % (range 61 – 93 %),
(Matthijs et al., 2004). Removal of alcohol ethoxylates in
anaerobic sludge of a single STP was also determined in
the range of 54 – 74 % by Bruno et al. 2002. Alcohol ethoxy-
lates were also detected by Cantero et al. in anaerobic sludge
at concentrations of 23 – 141 mg/kg while nonyl and octyl
phenol ethoxylates were in the range 11 – 151 and 100 – 138,
respectively. The removal of alcohol ethoxylates’ homologues
was > 99 % in nine US sewage treatment withactivated
sludge process. The total average concentration of alcohol
ethoxylates in effluent was 0.9 lg/l compared to 15.6 lg/l
in trikling filter (Morral et al., 2006). A monitoring of envir-
onmental fingerprints of alcohol ethoxylates in Europe and
Canada was published by Eadsforth et al. 2006.
No systematically investigations into the influence of the
branching degree of the alcohol moiety on the anaerobic bio-

Tenside Surf. Det. 44 (2007) 6 331

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review

degradability of alcohol ethoxylates are available. Mller
(Thesis, 2000) published some results of linear, single- and
multiple-branched alcohol ethoxylates using sludge from
different digester plants. The ethoxylation degree ranged
from 5 to 20 mol EO, which did not influence significant
the biodegradability. High degradation rates were found for
linear alcohol ethoxylates. Overall, the anaerobic biodegrad-
ability of alcohol ethoxylates decreased with increasing
branching degree of the alcohol moiety from linear to multi-
ple-branched alcohols.
Soil
There are no published relevant data of alcohol ethoxylates
in soil. Alcohol ethoxylates were detected at 194 ng/l in soil
interstitial water and 710 ng/l in ground water by Krogh
et al. 2002.
2530 mg/kg DM) were measured in digestors (Tschui and
Brunner, 1985).
2) Monitoring data
A complete review of the environmental fate of alkylphenols
and alkyl phenol ethoxylates was published by Guang-Guo
Ying 2002. APEs are biodegradable in aerobic and anaerobic
conditions but aerobic are preferred in fact, concentrations
of alkylphenol ethoxylates in the range < 0.1 to 13 700 lg/kg
were detected in sediments.
Low concentrations (35 – 95 mg/kg DM) of nonylphenol
in digester sludge have been reported (Kchler, 1995).
Sugar derivatives
1) Laboratory data

Sediments Alkylpolyglucosides (C8–-C12-linear alkyl) and glucoside

Alcohol ethoxylates were detected in ten sediments of Po riv- fatty acid esters are well degradable in anaerobic biodegrada-
er in the range 160 – 1600 lg/kg (Cavalli et al., 2000). tion screening tests (> 60 % biogas formation).
Alcohol ethoxylates and nonylphenol ethoxylates were Glucose Amides showed high mineralisation rates
detected in marine sediments of different Spanish coasts (> 80 %) in a 14C-simulation test.
and harbours in the range of 37 – 1300 lg/kg and 8 –
1080 lg/kg, respectively (Petrovic et al., 2002). More recently 2) Monitoring data
levels of alcohol ethoxylates within the same order of magni- No data available.
tude than LAS were found in the Bay of Cdiz area (Lara
Martin et al., 2006). Alcohol ethoxylates were also detected Amine oxides
in three US river sediments in the range 171 – 919 lg/kg,
most of these were represented by free alcohols (Dyer et al., Amine oxides are likely to be anaerobically degradable (1 po-
2006). sitive data point – DDAO).

Alkylphenol ethoxylates Alkyl ethanolamides

1) Laboratory data
Alkylphenol ethoxylates showed poor to moderate minerali-
zation rates (20 – 50 % biogas formation) in screening and
simulation tests.
Simulation tests indicated that degradation proceeds via
de-ethoxylation to alkylphenol which is poorly degradable.
Consequently, high concentrations of nonylphenol (up to
The anaerobic biodegradability of cocomonoethanolamide
has been examined by using the ISO 11 734 method. Ulti-
mate anaerobic biodegradation reached 81 % after 56 days
at 35 8C (Madsen, 2001).
7.3 Cationics – Amphoterics

Surfactant Characterisation Test Type Test Conc. Inocu- Test Temp. Results Remarks References
Type lum Dur-
Conc. ation
Screen- Simu- mg/l mg/l g/l days 8C %
ing lation active carbon
matter
Cationics
DTDMAC Dimethyl-di(14C) 14C-di- 10 *8 * 20 28 35 8.2 degradation Steber (1991)
Stearyl-ammo- gester (as dry (CH4) + 6.7 due to impurity
nium chloride matter) (CO2) = 14.9 ?
(C18)
DHTDMA- Hydrogenated ECETOC check not 35 No degrada- Garcia et al.,
C Tallow-based known tion (2000)
STAC 14C1-StearylTri- 14C-di- 0.98 - ww- 87 22 0 Federle and
Methyl ammo- gester mg/kg pond Schwab (1992)
nium chloride sedi- sedi-
(C 18) ment ment
500 -
ml/l
CTMAB Cetyltrimethyl Mod. 50 50 2 – 3 gl 60 35 inhibition Battersby and
ammonium bro- Shelton & Wilson (1989)
mide (C 16) Tiedje
MTEA Esterquat ECETOC 50 1 to 5 42 35 101.1 ± 12.8 Puchta et al.,
Esterquat (as dry (1993)
matter)

332 Tenside Surf. Det. 44 (2007) 6

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review

Surfactant Characterisation Test Type Test Conc. Inocu- Test Temp. Results Remarks References
Type lum Dur-
Conc. ation
Screen- Simu- mg/l mg/l g/l days 8C %
ing lation active carbon
matter
DEEDMAC Esterquat ECETOC * 38 1 to 5 60 35 90 Giolando et al.,
(C 16 – 18) (1995)
BAHMA Bis (acyloxy- ISO 50 not 120 25 24 Fraunhofer,
ethyl)-hydroxy- 11 374 known (Low number (2003)
methyl-methyl possibly due
ammonium to inhibition )
methosulfate
QFAI Quaternized fatty ISO 50 not 120 35 64 Fraunhofer,
acid imidazoline 11 374 known (2003)
methosulfate
Ampho-
terics
Betaines Coco Betaine ISO 100 not 84 35 0 (indications of Fraunhofer
11 374 known (Inhibition ?) substantial pri- (2003)
mary degrada-
tion)
CocoAmidoPropyl ISO 50 not 56 35 75 Madsen and
Betaine 11734 known Petersen,
(2001)
CocoAmidoPropyl ISO 100 not 84 35 60 – 80 Also high Fraunhofer,
Betaine 11734 known primary (2003)
degradation
(> 99 %)
Alkyl am- Disodium co- ISO 56 35 78 % of Petersen and
phoace- coamphodiace- 11734 theoretical Andersen,
tates tate gas produc- (2004)
tion

Conclusions regarding anaerobic degradability of cationic These compounds represent, however, only small usage vol-
surfactants umes as compared to the compounds in the table 7.3.

Cationics 2) Monitoring data

1) Laboratory data
Anaerobic degradability results for cationic surfactants need
to be evaluated with caution, since they may be inhibitory to
anaerobic metabolism at low levels (i. e. low mg/l range).
Nevertheless, some apparent trends can be observed:
y Mono-alkyl or di-alkyl quaternary nitrogen compounds
with straight (C–C) alkyl chains are not anaerobically de-
gradable (e. g. TMAC, DTDMAC, etc).
y Esterified mono-alkyl or di-alkyl quaternary nitrogen
compounds are biodegradable (e. g. MTEA esterquats,
DEEDMAC, etc). After ester hydrolysis, both the fatty
acid as well as the alcohol cleavage products can be
further completely mineralized.
y Only a single study is listed on quaternary imidazolium
salts, which seems to indicate the potential for anaerobic
degradability.
Since only few compounds have been tested, it is diffi-
cult to draw general conclusions for cationics with a high de-
gree of confidence. The effect on the degradability of factors
such as the position of the ester bond, the type of substitu-
tion on the nitrogen atom, branching, etc., have not been
systematically investigated.
For some categories of cationic surfactants, such a benzal-
konium- or ethoxylated quaternary salts no data were found.
Some monitoring data for DTDMAC/DODMAC in sludge,
sediments and soils are e. g. provided in the EU Risk Assess-
ment Report on DODMAC (2002).
There is no indication of DTDMAC being substantially
degraded in anaerobic sediments, or other anaerobic com-
partments.
STAC was not mineralized in anaerobic sediments of a
laundromat pond (Federle & Schwab, 1992).
These results are in line with the conclusion from labora-
tory tests on these chemicals.
Conclusions regarding anaerobic degradability of amphoteric
surfactants
Amphoterics
Laboratory data
The only categories of amphoteric surfactants for which data
were found are betaines and amidobetaines.
y Alkylbetaines: There remains uncertainty about the ulti-
mate anaerobic degradability of alkyl betaines. In the re-
ported experiment a high degree of primary degradation
was observed, but no apparent mineralization.

Tenside Surf. Det. 44 (2007) 6 333

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review
y Alkylamidobetaines are anaerobically well biodegradable.
y Alkylamphoacetates are also ultimately biodegradable
under anaerobic conditions.
No data were found for other groups of low volume am-
photerics such as glycinates, aminopropionates, sultaines, etc.
8 Evaluation of the Relevance of Anaerobic Biodegradation
8.1 Understanding mass fluxes of surfactants in environmental
compartments
A simplified scheme is used here to illustrate the typical
fluxes of surfactants through a Sewage Treatment Plant
(STP), and into different environmental compartments
(Fig. 7). The mass fluxes are driven by the following factors:
y The adsorption of surfactants on primary solids, which
will determine the fractions that go to aerobic and anae-
robic zones of a STP, respectively,
y The degradation extent of the surfactant in these two
zones taking into account the typical residence time
(6 h for the aeration tank and up to 3 weeks for the anae-
robic digester),
y The degradation extent in the receiving environmental
compartment, assuming a typical residence time (1 year
in agricultural soil, 13 years in landfill).
100 % sewage treatment, in line with the objectives of
the EU Urban Wastewater Treatment Directive. (N.B: a di-
rect discharge scenario of raw sewage is not included here
since other factors such as ammonium toxicity and oxygen
depletion will have a major environmental impact that will
override any potential surfactant effect). Besides, the scenar-
io is still conservative since it does not consider biodegrada-
tion on the sewers (Up to 60 %; Moreno and Ferrer, 1990
and Matthijs and Debaere, 1995).
Statistics from ADEME (1999) and Witte (2001) on the
disposal routes of sewage sludge in the EU were used to de-
fine the average fraction of sludge disposed to agricultural
land & forestry (50 %), landfill (25 %), incineration (20 %)
and other outlets (5 %). The other fraction covers minor
routes such as e. g. composting, pyrolysis and wet oxidation.
334
To illustrate the general behaviour and fluxes of surfac-
tants with different degradability patterns, two approaches
were followed: a modelling approach for three hypothetical
surfactants and an approach based on monitoring data for
real surfactants. Both approaches are complementary be-
cause data availability is extensive for LAS but less so for
other surfactant types.
8.1.1 Mass fluxes obtained via modelling for hypothetical surfactants
In the modelling approach Case 1 the surfactant is both aero-
bically and anaerobically degradable, whereas in Case 2 the
surfactant is aerobically but not anaerobically degradable.
The surfactant in Case 3 is neither aerobically nor anaerobi-
cally degradable. It should be noted that a Case 3 surfactant
would not be allowed in the market place today, according to
the EU Detergent Regulation (648/2004/EC), but the example
is used here to illustrate the effect of biodegradability).
In order to derive values for the various fluxes and to
reach conclusions with regard to the amount of surfactant
present in the various anaerobic compartments the follow-
ing working assumptions were made.
y 100 % of the surfactant used enter a STP. The potential
degradation that may have occurred already in the sewer
is not taken into account.
y 25 % of surfactant are removed by adsorption on primary
solids. The remaining 75 % go to aerobic treatment,
being consistent with a Kd of ca. 2000 l/kg).
y For surfactants that are readily (aerobically) degradable
(consistent with a primary degradation rate of 3/h) it is
assumed that
– 98 % are removed in a STP
– 95 % are removed in landfill
– 99 % are removed in agricultural soil (consistent with
a primary half life < 10 days)
y For surfactants that are also anaerobically degradable, it
is assumed that 80 % are removed by anaerobic digestion
y Since the disposal of sludge to the sea is not allowed any-
more, this route was not included
N.B: As noted in Figure 7, a) Flux 1 does not take into
account potential biodegradation in the sewer, and b) some
STPs have the capacity for aerobic stabilisation or compost-
* Flux N06 (sludge for agricultural use) may
undergo aerobic stabilisation prior to use on
land
Figure 7 Schematic representation of sur-
factant flux through a WWTP and final disposal
Tenside Surf. Det. 44 (2007) 6
J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review

ing of the sludge after anaerobic digestion. This process monitoring data for other surfactants are available (see ap-
would remove more than 90 % of aerobically degradable sur- pendix), the information does not allow to complete all
factants. As a result the numbers in Table 2 can be seen as a fluxes.
conservative scenario. The purpose of these examples is to Giger et al. (1989) investigated surfactant fluxes in the
illustrate the general behaviour and fluxes of surfactants Zurich STP and found LAS removal in the primary settler
with different degradability patterns rather than making a to be 27 %. Berna et al. (1989) doing similar work on a
realistic exposure evaluation on a country per country basis. WWTP in Madrid found 16 % removal, whereas Prats et al.
The model demonstrates how similar/different cases 1 and 2 (1997) working on the WWTP of Alicante found this fraction
are and what the impact is of the anaerobic biodegradation to be 37 %. The discrepancy between the latter case and the
on the fluxes. other two is explained by the unusually high water hardness
From Table 2 it becomes apparent that provided a surfac- in the Alicante region, and consequently the strongly de-
tant is readily (aerobically) biodegradable, even if it is not creased solubility of LAS in sewage (cf. 8.2.1). It seems that
anaerobically degradable (case 2), less than 1 % of the surfac- a removal of 25 % of LAS in the primary stage of a STP
tant released to a STP will reside in the permanently anaero- would be a fair estimate. This would mean that 75 % of the
bic compartments of a landfill, and the occasionally anaero- LAS entering a STP will go to aerobic treatment, with that
bic compartment of agricultural lands. For comparison, a on primary solids going to the anaerobic digester.
surfactant that is both aerobically and anaerobically biode- In an anaerobic digester, Prats et al. (1997) found 18 %
gradable (case 1) will show equilibrium levels that are removal of LAS, while Giger et al. (1987) found 20 – 30 %
5 times lower, approximately. and Osburn (1986) 0 – 35 % removal.
However, in the case of an aerobically and anaerobically Although degradation tests under strictly anaerobic cir-
non degradable surfactant (case 3), the percentage present cumstances indicate that sulfonates are not anaerobically de-
in agricultural land may be estimated to be at least 10 % of gradable, studies on these materials show extensive degrada-
the yearly flux. This is not only 100 times higher than in tion in case there has been a short exposure to oxygen. The
case 2, but additionally it must be expected that the concen- removals found in the monitoring studies may therefore re-
trations will increase continuously and may eventually cause sult from that fraction of LAS entering the digester which
ecotoxicological problems. At the same time, the flux which had been pre-exposed to aerobic conditions. In addition to
may end up sorbed onto river sediments is more than this, in some WWTPs some anaerobic sludge is returned to
30 times higher, again with the danger of increasing envi- the primary settler prior to aerobic composting or discharge
ronmental concentrations. It is for these reasons that the to the chosen sludge disposal route. To simplify matters it
EU legislation prescribes rapid aerobic degradability, and seems reasonable to take 15 % removal of LAS by anaerobic
that surfactants which are not aerobically degradable are treatment as a fair estimate. This percentage of removal has
not in use by the detergent industry. not been used in the modelling approach.
Prats et al. (1997) found that an aerobic stabilisation
8.1.2 Mass fluxes based on monitoring data step – where applied – removes a further 65 % of the re-
maining LAS. The sludge may be disposed of to agricultural
A similar balance of mass fluxes may be drawn up for con- land, to landfill, to the marine environment (practice ended
crete detergent surfactants where monitoring data are avail- in 1998) or to incineration. The same authors reported LAS
able. The environmental fate of LAS has been most exten- removal values of nearly 100 % using specific analytical
sively studied since it is one of the most widely used methods during composting of anaerobic sludge in a com-
surfactants and easy to determine analytically. Although mercial composting plant (Prats et al., 1999).
For the LAS fraction going to the aerobic process of a
Case 1 Case 2 Case 3 sewage treatment plant, Prats et al. (1997) found the amount
(%) (%) (%) leaving in effluent to be 0.6 % of that entering the STP,
Aerobic + Aerobic + Aerobic – equivalent to 99.4 % removal by the aerobic treatment. Rapa-
Anaer. + Anaer. – Anaer. –
port and Eckhoff (1990) found 98 % removal by activated
Flux 1 (input) 100 100 100 sludge plants although this was slightly lower for trickling
Flux 2 (to aerator) 75 75 75
filter plants. Feijtel et al. (1995) found an average of 99.2 %
removal when monitoring STPs in the Netherlands. In
Flux 3 (to digester) 25 25 25 agreement with the HERA risk assessment for LAS (HERA,
Flux 4 (to river) 2.0 2.0 75.0 2004) a figure of 99 % removal is a reasonable average,
hence, resulting in release of 1 % to receiving waters and
Flux 5 (from digester) 5 25 25
their sediments.
Flux 6 (to agriculture) 2.5 12.5 12.5 Holt et al. (1989) recorded in excess of 98 % LAS removal
% present in agricultural soil 0.025 0.125 > 12.5 *
in sludge-amended soils over a year, and calculated a half-
after 1 year life of 7 – 22 days. Ferrer et al. (1996) found 89.2 % removal
within 62 days giving a half-life of 19.3 days, while Kchler
Flux 7 (to incineration) 1 5 5
and Schnaak (1997) determined a 7-day half-life of LAS in a
Flux 8 (to landfill) 1.25 6.25 6.25 field trial study during the growth season.
Marcomini et al. (1988) monitored a landfill site having
% present in a landfill after 0.0625 0.31 > 6.25*
13 years received sewage sludge and found at the surface of the site
where sludge application was most recent, an LAS concen-
Flux 9 (to other) 0.25 1.25 1.25 tration of 9160 mg/kg while near the bottom this had
* The amounts cannot be accurately estimated because no equilibrium is reached, dropped to 245 mg/kg. This equates to an observed removal
Hence the given value is considered as a minimum, and a continuous build-up in of 97 % over a 13-year period.
this compartment can be assumed. There have been additional monitoring studies on other
Table 2 Modelled mass flux of hypothetical surfactants through a STP, and surfactant types but not as extensive as those for LAS. Prats
final concentrations present in a landfill and in agricultural soil et al. (1997) also followed the fluxes of nonionic surfactants

Tenside Surf. Det. 44 (2007) 6 335

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review

through a STP. However, whereas in their study of LAS this
material was followed specifically by HPLC, nonionics were
analysed using the Wickbold method (BiAS), a technique
which is not very specific and, hence, may also measure
non-ionic materials other than nonionic surfactants.
Field et al. (1995) studied the flux of secondary alkane
sulfonates (SAS) through a STP, allowing to construct the
mass balance in Table 3.
From all these studies approximate conclusions can be
drawn as to the amount of surfactant leaving WWTPs and
entering various environmental compartments, but there
are insufficient data available to assess their removal in
these compartments in great detail. It is however possible
to fill these gaps using circumstantial evidence. Since SAS,
like LAS, is aerobically but not anaerobically degradable, one
can assume comparable removals in landfill and agricultural
land of 98 and 97 %, respectively. Alcohol ethoxylates (AE)
are aerobically and anaerobically degradable and so removals
of 99 % could be expected in these compartments.
Alcohol ethoxy sulfates (AES), alcohol sulfates (AS), AE
and soap were also monitored in Dutch STPs (Feijtel et al.
(1995) using a highly specific analytical method. Based on
measurement of the influent and effluent concentrations,
removals of > 99 % were determined, corresponding to a flux
to rivers of 0.2 % (AE), 0.4 % (AES), 0.8 % (LAS, AS) and
0.95 % (soap), respectively. It is worth to note that the meas-
ured Biochemical Oxygen Demand (BOD) flux to rivers was
1.9 %, i. e. the surfactants were removed to a greater extent
than the sewage, and at least as efficiently as soap.
From the above monitoring data on these surfactants, a
mass fluxes table was drawn up (Table 3).
These monitoring data are in general agreement with the
modelled scenario of surfactant mass fluxes, indicating a
good conceptual understanding of surfactant behaviour in
the environment.
According to the values in Table 3, the level of an anaero-
bically degradable surfactant (e. g. AE) in soil after 1 year will
be 6 – 8 times lower than that of one which is only aerobi-
cally degradable (e. g. LAS). It illustrates that surfactants
which are readily biodegradable under both aerobic and
anaerobic conditions should have overall a lower potential
to cause an impact on soils and terrestrial biota.
AE [b] LAS
Flux 2 (to aerator) 44 % 75 % [b, e, f]
Flux 3 (to digester) 41 % 25 % [b, e, f]
Flux 4 (from digester) 14 % 21.3 % [b, e, g]
Flux 5 (to river) 0.2 % [d] 1.0 % [b, d, h]
Flux 6 (to agriculture) 1.6 % 7.7 %
% present in agricultural 0.02 % 0.15 % [i]
soil after 1 year
Flux 8 (to landfill) 2.1 % 9.9 %
% present in a landfill after 0.02 % 0.3 % [j]
13 years
Data in italics and with a shaded background are calculated from assumptions
given earlier in the text. The references provide the specific data sources.
[a] Field et al. (1995), [b] Prats et al. (1997),
[d] Feijtel et al. (1995), [e] Giger et al. (1989),
[f] Berna et al. (1989), [g] Osburn (1986),
[h] Rappaport and Eckoff (1990), [i] Holt et al. (1989),
[j] Marcomini et al. (1988)
However, the fact that a surfactant is anaerobically de-
gradable does not exclude its presence in anaerobic environ-
mental compartments as shown by field monitoring (see
8.2.1).
8.2 Impact of surfactants on structure and function
of anaerobic environmental compartments
8.2.1 Speciation and bioavailability of surfactants in anaerobic
compartments
The risk assessment of surfactants typically assumes that so-
luble species (usually Na salt; most bioavailable and biode-
gradable forms) are present in the environment. In analyti-
cal determinations of surfactants in soil, sediment and
sludge samples, the soluble form is created during the sam-
ple preparation. Nevertheless, in reality surfactants can be
subject to complex sorption and speciation processes (Ref
RISICO).
The speciation of surfactants present in anaerobic com-
partments has never been established exactly. The tendency
towards precipitation of surfactants, particularly anionics,
when reacting with water hardness (Ca++, Mg++) ions consti-
tutes together with adsorption and absorption phenomena
the most important mechanism ruling the presence of sur-
factant in environmental solid matrices. Such phenomena
might also occur with cationics as a consequence of the in-
teraction with various anionic species.
Understanding speciation of surfactants in anaerobic
compartments poses a scientific challenge, but would be a
valuable tool to assess the bioavailability and toxicity behav-
iour. There are, for example, several indirect indications of
the presence of LAS in solid environmental samples under
the form of insoluble Ca–Mg derivatives. While total surfac-
tant concentrations are sometimes high, the expected toxi-
city is not always observed, probably due to a low bioavail-
ability.
The solubility of Ca – LAS homologues in water is very
low. For C12 LAS the solubility at 250 mg/l water hardness
is in the order of 9 mg/l. This noticably influences the acute
toxicity to Daphnia Magna in such a way that, as the precipi-
tation progresses, the toxicity of LAS diminishes even at
high nominal concentration (Verge et al., 1997).
A clear relationship between LAS concentrations in pri-
SAS [a] mary sludge as well as on anaerobically digested sludge
84.4 %
and water hardness has been observed (Berna, 1989). A
comparison of the mass balance of Ca++ and Mg++ ions in
14.6 % the raw sewage, treated water and digested sludge indicates
16.3 % a remarkable accumulation of those ions in the sludge (Ber-
na, 1992). While the concentration of Ca++ + Mg++ is nearly
0.3 %
half of the amount of Na+ in raw sewage as well as in treated
5.9 % water, in the final sludge the concentration of Ca++ + Mg++ is
0.12 %
almost 30 times higher than [Na+].
A remarkable observation is that anaerobic digestion is
not inhibited by the presence of high levels of LAS, even
7.6 %
above the known inhibitory level of the Na-salt (Painter and
0.23 % Mosey, 1992; see also section 8.2.2.1). This is attributed to a
limited bioavailability. Hence, speciation aspects are impor-
Na+ Ca++ Mg++
Raw sewage (mg/l) 63 28 9
Treated water (mg/l) 60 26 8
Sludge (mg/kg) 1,200 21,500 9,000

Table 3 Estimated mass flux of high-volume surfactants based on monitor- Table 4 Comparison of concentrations of Na+, Ca++ and Mg++ in raw
ing data sewage, treated effluent and in sludge (Berna 1992)

336 Tenside Surf. Det. 44 (2007) 6

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review
tant to be considered when assessing the realistic ecotoxico-
logical impact of surfactants.
The same situation is most likely applicable to all anionic
surfactants. There are several publications showing consid-
erably high soap concentrations in anaerobic compartments
such as anaerobic sludge (Moreno et al., 1993) and marine
sediments (Folke et al., 2003). The anaerobic biodegradation
of soap in laboratory scale digesters as well as in full scale
plant is around 80 % (Prats et al., 1999). The low availability
and slow dissolution of Ca–Mg soap account for the less
than complete degradation.
While LAS present in those environmental compart-
ments can only be linked to its use in man-made detergent
products, the origin of soap, at least in compartments show-
ing extremely high concentrations like marine sediments, is
not yet clearly understood. The possibility of biochemical re-
actions involving fats and oils from a variety of (natural) ori-
gins taking place in the environment cannot be excluded.
For risk assessment purposes the total bioavailable concen-
tration of soap should be considered regardless of its origin.
8.2.2 Impact on wastewater treatment and aquatic environments
8.2.2.1 LAS and other anionic surfactants in anaerobic digesters
There is relatively little published information available on
the inhibitory effects of surfactants on anaerobic digestion.
Bruce et al. (1966) reported results obtained from laboratory
digesters fed on a daily basis with surfactant-free raw sludge
which had been amended with different concentrations of
surfactant. LAS (C 10–C 13, sodium salt) at a concentration
of 15 g/kg dry solids was not inhibitory; although biogas pro-
duction was reduced over the first 24 h after feeding, total
gas evolution after 48 h was similar to that in controls fed
with surfactant-free sludge only. At LAS levels between 15
and 20 g/kg dry solids the reliability of digestion was im-
paired and more serious inhibition occurred at higher con-
centrations. These findings were confirmed by Metzner
(1998), who found inhibitory effects on gas production at
ca. 1.1 g LAS/l resp. 22.5 g LAS/kg dry solid residue at a
dry solid concentration of 50 g/l. Similar results were ob-
tained for SAS by Bruce et al. (1966). C 16 – C 18 alcohol sul-
fate from natural raw materials as well as a petrochemical
based primary alcohol sulfate had both only a minor short-
term inhibitory effect on gas production at concentrations
up to 40 g/kg dry solids. At the end of the 18 day incubation
period, gas production in the test digesters was higher than
in the controls and 90 % of the surfactants had been de-
graded (MBAS removal). Nonylphenol ethoxylate had only
a slight inhibitory effect on gas production at levels up to
even 200 g/kg dry solids.
Published concentrations of surfactants in digester
sludge show that the majority of the data are for LAS, with
average levels around 5 g/kg dry solids (Schowanek et al., in
press). Hence, LAS levels in sewage sludge at most sewage
treatment works are below inhibitory levels. The toxicity of
LAS to anaerobic methanogenic bacteria was studied using
standard methods (Sanz et al., 1999). The experiments were
conducted on UASB (Upflow Anaerobic Sludge Blanket)
type reactors with a working volume of 315 mL. The 50 %
inhibition concentration (IC50) of the methanogenic activity
was reached at concentrations higher than 25 mg/l of LAS
(in the range of 40 to 150 mg/l) in all cases. These figures
are in agreement with other literature data, and are higher
than what is typically found in domestic wastewater.
The anaerobic digestion of AS-rich textile wastewater
(e.g, from cotton desizing) was tested in batch experiments
Tenside Surf. Det. 44 (2007) 6
(Feitkenhauer & Meyer, 2002). In the first part of the study
C10 AS was used as model to study the influence of concen-
tration in the potential inhibitory effects. The starch hydro-
lysis was inhibited at concentrations above 65 g AS/kg CDW
(cell dry weigh). In the second part of the study the inhibi-
tory effects was shown to be higher for medium and long
chain AS. The concentrations above which the inhibitory ef-
fects are observed are high. In domestic STPs no inhibition
attributed to AS has been reported.
Similar effects have been found before in experiments
with long chain fatty acids (soaps; Angelidaki and Ahring,
1992). More recently, the performance of laboratory scale
UASB and DASB reactors in the anaerobic biodegradation
of synthetic wastewater containing sodium oleate and so-
dium stereate was tested. The major problem encountered
was the adsorption of soap onto the solid support, impeding
therefore biomass-substrate contact and affecting the overall
biodegradation performance (Miranda et al., 2006).
8.2.2.2 Other surfactants in anaerobic digesters
The concentration of linear alcohol ethoxylate (AE) in diges-
ter sludge is around 0.5 g/kg dry solids, i. e. an order of mag-
nitude lower than LAS (Klotz, 1998). AE concentrations of a
comparable level were reported by Cavalli and Valtorta
(1999) and by Bruno et al. (2002) (< 700 and 312 mg/kg, re-
spectively) while Matthijs et al. (2004) detected AE concen-
trations in the range of < 22 – 468 (mean 168) mg/kg in the
digestor sludges from several European STPs. These AE
concentrations measured in practice can be compared with
the AE concentrations shown to be non-inhibitory in anaero-
bic degradation screening tests in the laboratory (cf. data in
chapter 7). The unrealistically high surfactant to biomass ra-
tio in these tests represents a worst case. Therefore inhibi-
tory effects in practice can be excluded if the surfactant con-
centration in practice does not significantly exceed the (non-
inhibitory) screening test concentration. A positive screen-
ing test result was obtained by Fraunhofer (2003) with a test
concentration of 120 g AE/kg sludge DM, i. e. more than
2 orders of magnitude higher than realistic AE concentra-
tions. Hence, inhibitory effects of AE on the function of
anaerobic digestion can be excluded.
Bruno et al. (2002) reported concentrations of other sur-
factants in digester sludge of the Roma Nord STP (Italy), re-
ceiving mainly dometic sewage. The following concentra-
tions were determined in 24-h composite samples of
digested sludge: 50 mg AS /kg dw and 69 mg AES /kg dw.
These can be compared with non-inhibitory test concentra-
tions of 96 g/kg for AS (Salanitro & Diaz, 1995) and ca.
17 g/kg for AES (Fraunhofer, 2003), respectively, underlin-
ing again that the realistic concentration of these surfactants
in digester sludges are well below the inhibition threshold
critical for the functioning of anaerobic digesters.
Despite the widespread use of anaerobic digestion, the
operation of these installations is not known to be disturbed
by the presence of surfactants.
8.2.2.3 Septic tanks and decentralized treatment systems
Septic tanks are onsite (pre-) treatment systems for black
water or unsettled domestic sewage. They are not considered
anymore state-of-the-art systems for sewage treatment but
nevertheless are still in use in Europe and the US, particu-
larly in rural areas. Septic tank effluents can be sent to a
sewer system, a ditch, or may be allowed (not generally) to
percolate into the soil (i. e. into a tile field). The septic tank
itself is an anaerobic/microaerobic system while the tile
337
J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review
field in the immediate vicinity of the tank, is usually a
strictly anaerobic environment. Surfactants released via a
septic tank system are therefore exposed first to an anaero-
bic/microaerobic compartment without dilution. BOD/COD
removal is around 50 % on average (LEAF, 2006).
The main function of a septic tank is to act as a settling
tank for the solids in domestic sewage, and needs to be per-
iodically emptied (e. g. once per year). This sludge is treated
elsewhere, usually in a nearby treatment plant, or may be
(not generally allowed) disposed to land. The water phase
in the tank has a residence time up to 5 days. The sludge at
the bottom of the tank undergoes an anaerobic decomposi-
tion, with the formation of biogas and H2S. This digestion is
the second function of a septic tank. Since the temperature
in the tank is close to that in the surrounding soil (< 20 8C),
digestion typically occurs at a slow rate.
Most surfactants and detergent formulations have been
tested for septic tank safety, i. e. when discharged under nor-
mal usage condition they will not disturb the functioning of
the septic tank. The protocol for such tests includes a para-
meter related to anaerobic digestion and settling rate under
realistic circumstances. This is mostly done in the labora-
tory, but some validation on real septic tanks has been per-
formed. Also surfactants which are not anaerobically degrad-
able can be safely used in septic tanks. Data to support this
are available within the Industry member companies. Some
studies are also available in the literature on the fate and be-
haviour of non-degraded surfactants leaving the tank into a
tile field (McAvoy et al., 1994; Shimp et al., 1994; McAvoy
et al., 2002).
A recent trend in Europe is the increase of so-called de-
centralized sewage treatment plants, i. e. small installations
designed for individual houses or small communities (< 100
inhabitant equivalents). They offer a better treatment perfor-
mance than septic tanks. In the ERASM-sponsored LeAF Re-
port (2006) an overview is given of the incidence of technol-
ogy for small and decentralized sewage treatment plants on
the EU market. In contrast to septic tanks, however, most of
these systems are based on aerobic processes. As a conse-
quence, they will not be discussed further in this review.
8.2.2.4 Anaerobic river and lake sediments
Freshwater sediments are usually anaerobic as of as few mm
or cm depth, and stratified in terms of redox potential and
the metabolic processes that occur within these zones. In
rivers, the structure of the sediment will be affected by the
velocity and turbulence of the river flow, which in turn
causes the transport of sediment particles either by suspen-
sion or by sliding and rolling (Press and Siever, 1986). Anae-
robic freshwater sediments play an important role in the de-
composition of organic matter, and in the cycling of
nitrogen, sulphur and phosphorus. These processes will be
dependent on the redox potential of the sediment, and in-
clude nitrate reduction to either ammonia or nitrogen, sul-
fate reduction to sulphide and methanogenesis (Jones,
1982). Sediments can also act as sinks for chemicals which
sorb to sediment particles or organic matter. The sediment
is also home to a wide range of fauna, flora and micro-or-
ganisms, and perturbation of these organisms can have a
detrimental effect of the structure and function of the eco-
system (e. g. Burton and MacPherson, 1995).
Available measured LAS data in freshwater sediments
were reviewed by Cavalli et al., (2000). Typical LAS values
in sediments below sewage outfalls were found in the 0.5 –
5.3 mg/kg range with an arithmetic mean of 2.9 mg/kg (12
records, SD = €1.9). Homologue distributions were also
338
measured for some river sediment samples (Cavalli et al.,
2000) and the corresponding fingerprint was found to be
similar to that of sludge and soils.
The occurrence of alkyl sulfates and alkyl ethoxysulfates
in US river sediments has been reported by Sanderson et al.
(2006). Combined levels of AS/AES were detected in a pre-
sumed lower exposed sites in the range of 0.025 –
0.034 mg/kg (dw) while a presumed higher exposed site
had combined levels of 0.117 mg/kg dw.
Little information of the toxicity of LAS (or other surfac-
tants) to anaerobic, sediment bacteria could be found. Jen-
sen et al. (2001) report an EC10 (= surrogate of a NOEC) of
5 mg LAS/kg soil for the anoxic process of microbial iron re-
duction. Data on the toxicity of LAS to aerobic bacteria
(EC50 = 183 g/kg dw of activated sludge) (HERA, 2004) and
anaerobic digestion (EC0 > 15 g/kg, see 8.2.2.1) indicate that
the levels of LAS in river sediments are not expected to inhi-
bit microbial processes.
As regards higher organisms in sediments, Pittinger et
al. (1991) reported a NOEC = 319 mg/kg dry solids for the
larvae of a benthic organism, Chironomus riparius (midge).
In addition, subchronic toxicity tests with mussels (Unio
elongatulus), and worms (Lumbriculus variegatus, Caenor-
habditis elegans) revealed NOEC values of > 200, 81 and
100 mg/kg, repectively (HERA, 2004). Also these data show
clearly that the concentration of LAS in freshwater sedi-
ments is at least one order of magnitude below the levels
which might impact freshwater organisms.
Considering the prominent role of LAS in terms of the
usage volume in detergents and an ecotoxicological profile
similar to other relevant surfactants used in detergents, it
can be assumed that the above conclusion on LAS is also val-
id for the other detergent surfactants.
8.2.2.5 Agricultural soil (pasture and cropland)
About 50 % of the European sewage sludges are disposed on
agricultural land (Schowanek et al., 2004). A framework for
performing an ecological risk assessment for chemicals in
agricultural soil is provided in e. g. the EU TGD (2003) and
by ILSI-Europe (Schowanek et al., 2004). According to the
European Technical Guidance documents (EU-TGD, 2003)
for sludge application to agricultural soil, an application rate
of 0.5 kg sewage sludge DM/m2 per year is assumed
(= 5000 kg/ha.year). This sludge is mixed into 20 cm of top-
soil, with an average density of 1.3 tonne/m3. The EU TDG
approach also considers a 30-day time-weighted averaging
while calculating PECsoil, in order to account for the fact that
seeding/planting typically occur a few days/weeks after ap-
plication of the sludge.
Assuming an avarage European LAS concentration in
anaerobic sludge of 5000 mg/kg, (Schowanek et al., 2007),
the predicted initial soil LAS concentration is around
10 mg/kg immediately after sludge application3. Aerobic
conditions prevail in sludge-amended soils, as shown by sev-
eral studies which demonstrated that LAS disappears with
half lives of 7 – 30 days (mean value * 10 days) (De Wolf,
1998; Cavalli, 1998; Schowanek et al., in press). Due to the
averaging time concept the LAS concentration in soil (PEC-
soil) as used in EU risk assessment is less than half the
expected initial value of 10 mg/kg, i. e. below 5 mg/kg.
No field monitoring has been performed to determine
PEC soil for LAS or any other surfactants under the condi-
3
This value is often lower since (anaerobic) sewage sludge is often stored and
stabilized for a few weeks or months before use in agriculture. During this
period aerobic biodegradation will lower the surfactant concentration.
Tenside Surf. Det. 44 (2007) 6
J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review
tions (dosage, mixing, time average, . . . etc.) described in the
EU TGD. Nevertheless, many monitoring studies on LAS
confirm the general degradation behaviour of the molecule,
and importantly, the absence of accumulation behaviour
over time.
ILSI-Europe has done an extensive review and probabil-
itic risk assessment for LAS in sewage sludge used on agri-
cultural soil, looking at a variety of relevant endpoints (Scho-
wanek et al., 2004, Schowanek et al., in press). This study
concludes that the presence of LAS in sludge poses no risk
to agricultural soil and the foodchain, up to very high levels
(i. e. 55 and 27.5 g LAS/kg sludge for a homogeneous mix-
ing and injection scenario, respectively). Such LAS levels
are not reached in European sewage sludge.
Laboratory studies carried out on sludge stored on com-
pacted soil floors demonstrate that LAS penetrated, even
after an extended time period of 1 year, only to a minimum
degree into the soil. Most of the LAS migration was found in
the 3 cm thick soil layer. No LAS was present in the water
percolating through the soil. Leaching of LAS from the
sludge amended soil to ground water should not be expected
to happen (Figge, 1991).
The leaching potential of LAS in agricultural soils has
also been investigated in soils irrigated with water contain-
ing measured LAS concentrations. No LAS was detected be-
low the top 10 cm soil layer after one year of periodical irri-
gations.
For surfactants other than LAS (i. e. AS, AES, SAS, soap,
esterquats and AE (in preparation), the respective HERA re-
ports (www.heraproject.org) provide a risk assessment based
on the available terrestrial toxicicity data. This provides a
consistent picture that at current usage volumes none of
these surfactants gives rise to concern for the terrestrial en-
vironment, regardless whether they are anaerobicically bio-
degradable or not.
It is sometimes suggested that surfactants, applied with
the use of sewage sludge in agriculture, might increase the
mobility of organic micropollutants. These effects, however,
were only detected at very high surfactant concentrations
which are not relevant in the agricultural practice (Kchler,
1995). This can also be explained by the fact that surfactants
applied via sewage sludge occur in soil in a form adsorbed to
sludge particles.
8.2.2.6 Landfills for sludge
Sanitary landfilling is widely used to dispose of diverse types
of waste, including digested and dewatered sludge. How-
ever, over the last years the trend in Europe has been to-
wards decreased landfilling and a corresponding increase
in incineration (Schowanek et al., 2004). Well run landfills
are isolated from the surrounding environment (ground-
water, air, rain). This isolation is accomplished with a bot-
tom liner system (clay, plastic foils) and daily covering of
soil.
Conditions inside a landfill may vary spatially and tem-
porarily between aerobic, anoxic and strictly anaerobic con-
ditions. Landfills are not designed to break down waste,
merely to bury it. However, for organic wastes with asuffi-
cient moisture content like sludges from wastewater treat-
ment plants a landfill may behave similarl to a bioreactor
providing suitable conditions for anaerobic bacteria. This
will lead to the conversion of organic wastes into organic
acids and ultimately into methane and carbon dioxide (bio-
gass), which is often recuperated for energy generation.
Since landfills are not normally opened there is only
scarce information about surfactant concentrations in land-
Tenside Surf. Det. 44 (2007) 6
fills. One study (Marcomini, 1989) gives data on TPS (Tetra-
propylenebenzene Sulfonate) and LAS for four German
landfills, which were used as deposits for digested sludge
for up to 30 years. The switch fromTPS to LAS can be used
as a time marker to help to set the age of the landfill. Con-
sidering that LAS consumption, and consequently its input
to landfills, has only slightly increased along the years taken
into account, it appears that, with a residence time of
13 years, the LAS concentration decreased from about 9 g/
kg to 0.2 g/kg in one landfill. This decrease may rather be
indicative of anoxic conditions (oxygen limitation) in the re-
spective landfill allowing a slow aerobic degradation of LAS.
There is no evidence of disturbance of landfill function-
ing by surfactants. Lack of anaerobic biodegradability is not
considered relevant in landfills.
8.2.2.7 Marine sediments
The structure and function of marine sediments are compar-
able similar to that of freshwater sediments, although there
are some key differences. In estuarine and coastal areas, se-
diments are usually anaerobic below the surface few mm
but pelagic sediments can be oxic at sediment depths > 1 m
(Jørgensen, 1982). The stratification of anaerobic decompo-
sition and nutrient cycling processes with decreasing sedi-
ment redox potential are similar to freshwater sediments.
However, a big difference is the abundance of sulfate in sea-
water which means that sulfate reduction replaces methano-
genesis as the terminal stage in the anaerobic oxidation of
organic matter. In coastal waters this process can convert as
much organic matter to CO2 as do aerobic organisms in
aerobic environments.
Monitoring data for concentrations of surfactants in es-
tuarine or marine sediments are scarce, and those data avail-
able are mostly confined to LAS. In their review on the en-
vironmental safety of LAS, Painter and Zabel (1988)
reported levels of 5 – 17 mg/kg dry solids in estuarine sedi-
ments close to the outfalls of probably untreated sewage.
LAS was undetectable further out to sea. The same trend
was observed in the study carried out in sediment samples
from the Baltic proper and Little Belt (Denmark) by Folke
et al. High concentration of LAS was measured in the inner
part of the shipping port (BABS, the branched alkylbenzene
used in the 50’s and early 60’s, was even detected). As sedi-
ments became cleaner with increasing distance from the
shipping ports, the levels of LAS decreased to near or below
the detection limit (0.1 mg/kg sediment DM) indicating
therefore that Danish marine sediments are not generally
contaminated with LAS. The aforementioned study also in-
cluded the monitoring of soap concentrations. The concen-
tration found in relatively uncontaminated sediment was
15 000 – 40 000 times higher than that of LAS. High concen-
trations of soap in sediment (1000 – 2000 mg/kg DM) were
observed in areas where LAS could not be detected
(< 0.05 mg/kg DM). The origin of all detected soap could
not be pinpointed, since soaps can also be formed by natural
processes.
The variation with depth of the concentrations of LAS
and of the long-chain sulfophenylcarboxilic acids (SPCs – re-
sulting from LAS biodegradation) was determined in coastal
sediments (Len et al., 2001). The average surface layer con-
centration (0 – 8 cm depth) of total LAS varied a lot depend-
ing on the sample point, More than 100 ppm were detected
in the vicinity of the discharge point on non treated waste-
water (this is considered as non representative data) while
LAS levels ranged from values of several 10’s of ppb to few
ppm were found in cleaner areas. The vertical profile of LAS
339
J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review
concentration showed a sharp reduction with depth while
SPCs levels increased. The presence of SPCs in sediment
layers considered to be anoxic, together with the disappar-
eance of LAS, led the authors to postulate the existence of
some kind of mechanism for LAS degradation under anoxic
conditions in the marine environment.
The same research group from the University of Cdiz
studied further the distribution of LAS as well as AES in
the sediments taken from the same coastal area of Spain
(Gonzlez Mazo et al., 2005). A decrease in LAS concentra-
tions was found compared to past studies in one of the sam-
pling areas due to the entry into service of a STP six months
before the time of the sampling. With respect to the AES
concentration levels values ranging between 150 and
400 ppb were reported. These concentrations are lower than
those for LAS, especially near the effluent outlet, mainly due
to the lower AES consumption in Spain. More recently, an
additional monitoring study at the Bay of Cdiz was carried
out, this time extending the analysis with AEs (Lara Martn
et al., 2005). Concentrations were up to 637 ppb for LAS,
861 ppb for AEs and 125 ppb for AES in surface sediments.
It is worth noting that concentration of AE is slightly higher
than that of LAS, despite the fact the AE is known to be bio-
degradable under both aerobic and anaerobic conditions.
The main trend along the sediment columns is towards a
concentration decrease for of all these compounds with
depth.
There is a general paucity of data regarding the toxicity
of surfactants to marine sediment dwelling organisms. This
is an area that ERASM technical committee has started to
address and information should be available in the coming
years.
References
Ahel, M. and Giger, W.: Determination of alkylphenols and alkylphenol mono-and
diethoxylates in environmental samples by high performance liquid chromato-
graphy. Anal. Chem. 57 (1985) 1577 – 1583.
Ahel, M., Giger, W. and Koch, M.: Behaviour of alkylphenol polyethoxylate surfac-
tants in the aquatic environment – I. Occurrence and transformation in sewage
treatement. Wat. Res 28 (1994) 1131 – 1142.
Ahel, M., Schaffner, C. and Giger, W.: Behavior of Alkylphenol ethoxylate surfactants
in the Aquatic environment – III. Occurrence and elimination of their persistent
metabolites during infiltration of river water to groundwater. Wat. Res.: 30
(1996) 37 – 46.
AISE/CESIO monitoring on European sludges.
Amano, K., Fukushima, T. and Nakasugi, O.: Diffusive exchange of linear alkylben-
zenesulfonates (LAS) between overlying water and bottom sediment. Hydro-
biologia 235/236 (1992) 491 – 499.
Angelidaki, I. and Ahring, B. K.: Effect of long-chain fatty acids on thermophilic
anaerobic digestion. Appl. Microbiol. Biotechnol. 37 (1992) 802 – 812.
Angelidaki, J., Hagensen, F. and Ahring, B. K.: Anaerobic transformation of LAS in
continuous stirred tank reactors treating sewage sludge. In Proceedings of the
5th World Surfactant Congress. Comite Européen des Agents de Surface et
Leurs Intermediares Organiques. Milano, Italy 2000.
Angelidaki, I., Torang, L., Waul, C. M. and Schmidt, J. E.: Anaerobic bioprocessing of
sewage sludge, focusing on degradation of linear alkylbenzene sulfonates
(LAS). Water Science and Technology 49 (10, Wastewater Sludge as a Re-
source), (2004) 115 – 122.
ASTM E 1196-87. Standard Method for Determining the Anaerobic Biodegradation
Potential of Organic Chemicals. (1987) 738 – 742.
Austermann-Haun, U., Kroiss, H., Fischer, P., Joerdening, H.-J., Meyer, H., Pascik, I.,
Rosenwinkel, K.-H., Seyfried, C. F., Strotmann, U. J., Svardal, K., Temper, U.,
Weiland, P., Winter, J. and Metzner, G.: Fünften Arbeitsbericht der ATV-DVWK-
Arbeitsgruppe IG 5.1. KA – Abwasser, Abfall, 51 (2004) 997 – 1001.
Battersby, N. S. and Wilson, V.: Evaluation of a serum bottle technique for assessing
the anaerobic biodegradability of organic chemicals under methanogenic con-
ditions. Chemosphere 17 (1988) 2441 – 2460.
Battersby, N. and Wilson, V.: Survey of the anaerobic biodegradation potential of
organic chemicals in digesting sludge. Appl. Environ. Microbiol. 55 (1989)
433 – 439.
Baumann, H. and Schefer, W.: Simple test system to judge the anaerobic biode-
gradability of organic substances. Textilveredlung 25 (1990) 248.
Baumann, U. and Mueller, M. T.: Determination of anaerobic biodegradability with a
simple continuous fixed bed reactor. Wat. Res. 31 (1997) 1513 – 1517.
Berna, J. L., Ferrer, J., Moreno, A., Prats, D. and Ruiz, F.: The fate of LAS in the en-
vironment. Tenside Surf. Det. 26 (2) (1989) 101 – 107.
Berna, J. L., Moreno, A. and Ferrer, J.: An assessment of the ultimate biodegradation
of LAS. Proceedings of CESIO World Surfactants Congress, London (UK), 3
(1992) 59 – 72.
340
Bester, K., Theobald, N. and Schroeder, H. Fr.: Nonylphenols, nonylphenol-ethoxy-
lates, linear alkylbenzenesulfonates (LAS) and bis (4-schlorophenyl)–sulfone
in the German Bight of the North Sea. Chemosphere. 45 (2001) 817 – 826.
Birch, R. R., Biver, C., Campagna, R., Gledhill, W. E., Pagga, U., Steber, H., Reust, H.
and Bontinck, W. J.: Screening chemicals for anaerobic biodegradability. Che-
mosphere 19 (1989) 1527 – 1550.
Birch, R. R., Geldhill, W. E., Larson R. J. and Nielsen, A. M.: Proc. 3rd CESIO Interna-
tional Surfactants Congress and Exhibition, London, (1992) 26 – 33.
Bouwer, E. J. and McCarty, P. L.: Transformation of halogenated organic compounds
under denitrification conditions. Appl. Environ. Microbiol. 45 (1983) 1295 –
1299.
Bruce, A. M., Swanwick, J. D. and Ownsworth, R. A.: Synthetic detergents and
sludge digestion: Some recent observations. The Journal and Proceedings of
the Institute of Sewage Purification 5 (1966) 427 – 447.
Brunner, P. H., Capri, S., Marcomini, A. and Giger, W.: Occurrence and behaviour of
linear alkylbenzene sulfonates, nonylphenol, nonylphenol mono-and nonyl-
phenol diethoxylates in sewage and sewage sludge treatment. Wat. Res. 22
(1988) 1465 – 147.
Bruno, F., Curini, R., Di Corcia, A., Fochi, I., Nazzari, M. and Saperi, R.: Determination
of surfactants and some of their metabolites in untreated and anaerobically
digested sewage sludge by subcritical water extraction followed by liquid
chromatography-mass spectrometry. Environm. Science & Technology 36
(2002) 4156 – 4161.
Burton Jr., G. A. and MacPherson, C.: Sediment toxicity testing issues and methods.
In Handbook of Ecotoxicology, eds. D. J. Hoffman, B. A. Rattner, G. A. Burton Jr.
and J. Cairns Jr., 70 – 103. CRC Press Inc. (1995).
Cantero M., Soledad, R. and Pérez-Bendito, D.: Determination of non-ionic poly-
ethoxylated surfactants in sewage sludge by coacervative extraction and ion
trap Liquid chromatography-mass spectrometry. J. Chromatogr. A. 1046
(2004) 147 – 153.
Carlsen, L., Metzon, M.-B. and Kjelsmark, J.: Linear alkylenzenene sulfonatee (LAS)
in the terrestrial environment. Sci. Tot. Environ. 290 (2002) 225 – 230.
Casellato, S., Aeillo, R., Negrisolo, A. and Seno, M.: Long-term experiment on Bran-
chiura sowebyi Beddard (oligochaeta, tubificidae) using sediment treated with
LAS (Linear Alkylbenzene Sulfonate). Hydrobilogia 232 (1992) 169 – 173.
Cavalli, L., Gellera, A. and Landone: LAS removal and biodegradation in a waste
water treatment plant. Env. Tox. Chem. 12 (1993) 1777 – 1788.
Cavalli, L. and Valtorta, L.: Surfactants in sludge-amended soil, 1998, XXVIII CED,
Barcelona, May 6 – 8 (1998).
Cavalli, L., Cassani, G., Viganò, L., Pravettoni, S., Nucci, G., Lazzarin, M. and Zatta,
A.: (2000). Surfactants in sediments. Tenside Surf. Det. 37 (5) (2000) 282 –
288.
Cohen, A., Klint, K., Bowadt, S., Persson, P. and Jonsson, J. A.: Routine analysis of
alcohol and nonylphenol polyethoxylates in wastewater and sludge using liquid
chromastography-electrospray mass spectrometry. J. of. Chrom. A, 927 (2001)
103 – 110.
Compagnion, D. and Nyns, E.-J.: Biomethanation in developing countries. In Bio-
technology Vol. 8. Microbial Degradations, ed. W. Schönborn VCH, (1986)
239 – 267.
Chalaux, N., Bayona, J. M. and Albaiges, J.: Determination of nonylphenols as pen-
tafluorobenzyl derivatives by capillary gas chromatography with electron cap-
ture and mass spectromentric detection in environmental matrices. J. Chro-
matogr. 686 (1994) 275 – 281.
Chien Chih-Ching, Leadbetter, E. R. and Godchaux III, W.: Sulfonate-sulfur can be
assimilated for fermentative growth, FEMS Microbiology Letters, 129 (1995)
189 – 194.
DelValls, T. A., Forja, J. M. and Gomez-Parra, A.: Seasonality of contamination, toxi-
city, and quality values in sediments from littoral ecosystems in the Gulf of
Cadiz (SW Spain). Chemosphere. 46 (2002) 1033 – 1043.
DiCorcia, A., Samperi, R., Bellioni, A., Marcomini, A., Zanette, M., Lenr, K. and Ca-
valli, L.: LAS Pilot study at the “Roma-Nord” sewage treatment plant and in the
Tiber river. Riv. Ital. Sost. Grasse 71 (1994) 467 – 475.
Denger, K., Kertez, M. A., Vock, E. H., Schon, R. and Cook, A. M.: Anaerobic desul-
fonation of 4-tolylsulfonate and 2-(4-sulfophenyl) butyrate by Clostridium spp,
Appl. Environ. Microbiology 62 (1996) 1526 – 1530.
Denger, K. and Cook, A. M.: Assimilation of sulfur from alkyl- and arylsulfonates by
Clostridium spp, Arch. Microbiol. 167 (1997) 177 – 181.
Denger, K., Lane, H. and Cook, A. M.: Anaerobic taurine oxidation: a novel reaction
by nitrate-reducing Alcaligenes spp, Microbiology 143 (1997) 1919.
Denger, K. and Cook, A.: LAS bioavailable to anaerobic bacteria as a source of sulfur,
Appl. Environ. Microbiol. 86 (1999) 166 – 168.
Deuser, W. G.: Organic carbon budget of the Black Sea. Deep Sea Research 18
(1971) 995 – 1004.
De Wolf, W. and Feijtel, T.: TRA for LAS in sludge-amended soil, Chemosphere, 36
(1998) 1319 – 1343.
DiCorcia, A., Samperi, R., Bellioni, A., Marcomini, A., Zanette, M., Lenr, K. and Ca-
valli, L.: LAS Pilot study at the “Roma-Nord” sewage treatment plant and in the
Tiber river. Riv. Ital. Sost. Grasse 71 (1994) 467 – 475.
Dyer, S. D., Sanderson, H., Waite, S. W,. Compernolle, R. V., Price, B., Nielsen, A. M.,
Evans, A., Decarvallo, A. J., Hooton, D. J. and Sherren, A. J.: Assessment of al-
cohol ethoxylate surfactants and fatty alcohols mixtures in river sediments and
prospective risk assessment. Environ. Mon. & Assess. 120 (2006) 45 – 63.
Eadsforth, C. V., Sherren, A. J., Selby, M. A., Toy, R., Eckhoff, W. S., McAvoy, D. C. and
Matthijs, E.: Monitoring of environmental fingerprints of alcohol ethoxylates in
Europe and Canada. Ecotoxycol. and Environ. Saf. 64 (2006) 14 – 29.
ECETOC. (1988). Evaluation of anaerobic biodegradation. Technical Report 28.
EEA (European Environment Agency). Urban wastewater treatment. ETC/WTR
based on Member States data reported to OECD/EUROSTAT Joint Question-
naire 2002.
Eichhorn, P., Lopez, O. and Barcelo, D.: Application of liquid chromatography-elec-
trospray-tandem mass spectrometry for the identification and characterisation
of linear alkylbenzene sulfonates and sulfophenyl carboxylates in sludge-
amended soils. Journal of Chromatography A. 1067(1 – 2) (2005) 171 – 179.
Tenside Surf. Det. 44 (2007) 6
J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review
EPA EPA 40 CFR Ch. 1 (7-1-88 Edition) § 796.3140: Anaerobic biodegradability of
organic chemicals. (1988) 568 – 572.
EU-TGD. Technical Guidance Documents (TGD) in support of EU Directive 93/67
EEC and EU Regulation 1488/94 published by the EU Chemical Bureau, 1996.
Federle, T. W. and Schwab, B. S.: Mineralization of surfactants in anaerobic sedi-
ments of a laundromat wastewater pond. Wat. Res. 26 (1992) 123 – 127.
Fernandez, P., Alder, A. C., Suter, M. J. F. and Giger, W.: Determination of the qua-
ternary ammonium surfactant ditallowdimethylammonium in digested sludges
and marine sediments by supercritical fluid extraction and liquid chromatogra-
phy with post-colums ion-pair formation. Anal. Chem. 68 (1996) 921 – 929.
Feitkenhauer, H. and Meyer, U.: Anaerobic Digestion of alcohol sulfate (anionic
surfactant) rich wastewater –batch experiments. Part I: Influence of the sur-
factant concentration. Bioresource Technology 82 (2002) 115 – 121.
Feitkenhauer H. and Meyer U.: Anaerobic Digestion of alcohol sulfate (anionic sur-
factant) rich wastewater – batch experiments. Part II: Influence of the hydro-
phobic chain length. Bioresource Technology 82 (2002) 123 – 129.
Feijtel, T. C. and van den Plassche, E.: Environmental Risk Characterisation of four
Major Surfactants used in the Netherlands, Report No 679101025 by NVZ –
RIVM (1995).
Ferrer, J. de, Moreno, A., Vaquero, Ma. T. and Comellas, L.: Monitoring of LAS (Lin-
ear Alkylbenzene Sulfonate) in direct discharge situations: untreated sewage
and on sludges amended soils, CESIO 4th World Surfactant Congress, Barcelo-
na (Spain) (1996).
Field, J., Miller, D. J., Field, T. M., Hawthorne, S. B. and Giger, W.: Quantitative de-
termination of sulfonated aliphatic and aromatic surfactants in sewage sludge
by ion-pair/supercritical fluid extraction and derivatization GC/MS, Anal. Chem.
64 (1992) 3161 – 3167.
Field, J. A., Field, T. M., Poiger, T., Siegrist, H. and Giger, W.: Fate of secondary alkane
sulfonate surfactants during municipal wastewater treatment. Wat. Res. 29
(1995) 1301 – 1307.
Figge, K.: NATEC report, Fate of LAS in digested sewage sludge deposits, February 1991.
Folke, J., Cassani, G., Ferrer, J., López, I., Karlsoon, M. O. and Willumsen, B.: Linear
Alkylbenzene Sulfonates, Branched Dodecylbenzene Sulfonates and Soap
Analysed in Marine Sediments from the Baltic Proper and Little Belt. Tenside
Surf. Det. 40 (1) (2003) 17 – 24.
Fraunhofer. Anaerobic biodegradation of detergent surfactants. Final report to the
European Commission Enterprise Directorate-General, (July 2003). Fraunhofer
Institut Umwelt-, Sicherheits-Energietechnik UMSICHT, Oberhausen/Germany
2003.
Garcia, M. T., Campos, E., Illan, P., Dalmau, M. and Sanchez-Leal, J.: Study of the
anaerobic biodegradation of homologs and pure isomers of linear alkylben-
zene sulfonates in discontinuous digesters. Comunicaciones presentadas a las
Jornadas del Comite Espanol de la Detergencia 34 (2004) 147 – 160. Comite
Espanol de la Detergencia, Tensioactivos y Afines.
Garcia, M. T., Campos, E., Ribosa, I., Latorre, A. and Sanchez-Leal, J.: Anaerobic di-
gestion of linear alkyl benzene sulfonates: biodegradation kinetics and meta-
bolite analysis. Chemosphere 60 (11) (2005) 1636 – 1643.
Garcia, M. T., Campos, E., Sanchez-Leal, J. and Ribosa, I.: Effect of linear alkylben-
zene sulfonates (LAS) on the anaerobic digestion of sewage sludge. Water
Research 40 (2006) 2958 – 2964.
Garcia, M., Teresa, C., Encarna, D., Manel, I. P. and Sanchez-Leal, J.: Inhibition of
Biogas Production by Alkyl Benzene Sulfonates (LAS) in a Screening Test for
Anaerobic Biodegradability. Biodegradation, 17 (1) (2006) 39 – 46.
Giger, W., Brunner, P. H. and Schaffner, C.: 4-Nonylphenol in Sewage sludge: Accu-
mulation of Toxic Metabolites from Non-ionic Surfactants. Science 225, vol. 10,
August. (1984) 623 – 625.
Giger, W., Brunner, P. H., Ahel, M., McEvoy, J., Marcomini, A. and Schaffner, C.: Or-
ganic detergent components and their degradation products in wastewater
and sludge. Gas, Wasser, Abwasser 67 (1987) 111 – 122.
Giger, W., Brunner, P. H., Marcomini, A. and Siegrist, H.: Behaviour of LAS in sewage
and sludge treatment plant. Tenside Surf. Det. 26 (2) (1989) 95 – 100.
Giolando, S., Rapaport, R. A., Larson, R. J. and Federle, T. W.: Environmental fate and
effects of DEEDMAC: a new rapidly biodegradable cationic surfactant for use in
fabric softeners. Chemosphere 30 (1995) 1076 – 1083.
Gledhill, W. E.: Review of the anaerobic biodegradation test methodology: relevance
to the environmental anaerobic conditions and detergent environmental data
– A literature review for the US Soap & Detergent Association (1995).
González Mazo, E., Lara Martín, P. A. and Gómez Parra, A.: Determination and dis-
tribution profiles of Alkyl Ethoxysulfates and Linear Alkylbenzene Sulfonates in
coastal marine sediments from the bay of Cádiz (Southwest of Spain). Envir-
onmental Toxicology and Chemistry, Vol. 24 (9) (2005) 2196 – 2202.
Guang-Guo Ying, Williams, B. and Kookana, R.: Environmental fate of alkylphenols
and alkylphenol ethoxulates. Environ. Itern. 28 (2002) 215 – 226.
Haggensen, F., Mogensen, A. S., Angelidaki, I. and Ahring, B. K.: Anaerobic treat-
ment of sludge: focusing on reduction of LAS concentration in sludge. Water
Science and Technology, 46 (10, Sludge Management: Regulation, Treatment,
Utilisation and Disposal), (2002) 159 – 165.
Harrits, S. M. and Hanson, R. S.: Stratification of aerobic methane-oxidizing organ-
isms in Lake Mendota, Madison, Wisconsin. Limnology and Oceanography 25
(1980) 412 – 421.
Heijnen, J. J., Mulder, A., Weltevrede, R., Hols, J. and Van Leeuwen, H. L. J. M.: Large-
scale anaerobic-aerobic treatment of complex industrial wastewater using bio-
film reactors. Water Science Technol. 23 (7 – 9) (1991) 1427 – 1436.
Heinze, J. and Britton, L.: Anaerobic biodegradation: environmental relevance, Pro-
ceedings 3rd World Conference on detergents: global perspectives (ed.
A. Cahn), p. 235, AOCS Press, Champaign, IL (1994).
HERA. Human and Environmental Risk Assessment of Linear Alkylbenzene Sulfo-
nate. Available from <www.heraproject.com> (2004).
HMSO. The Assessment of Biodegradability in Anaerobic Digesting Sludge 1988.
Methods for the Examination of Waters and Associated Materials. Her Majesty’s
Stationery Office: London, 1989.
Holt, M., Matthijs, E. and Waters, J.: The concentrations and fate of LAS in sludge
amended soils, Water Research, 23 (6) (1989) 749 – 759.
Tenside Surf. Det. 44 (2007) 6
Holt, M. S. and Bernstein, S. L.: Linear alkylbenzenes in sewage sludges and sludge
amended soils. Wat. Res.; 26 (1992) 613 – 624.
Holt, M. S., Mitchell, G. C. and Watkinson, R. J.: The environmental chemistry, fate
and effects of non-ionic surfactants. In: the Handbook of Environmental
Chemistry , Vol. 3, Part F. Anthropogenic Compounds, Detergents. (ed. N. T. de
Oude), 89 – 144, Springer Verlag, Berlin, 1992.
Hon-Nami, H. and Hanya, T.: Linear alkylbenzene sulfonates in river, estuary and
bay water. J. Jap. Limnol 41 (1 – 5) (1980).
Howarth, R. W. and Teal, J. M.: Sulfate reduction in a New England salt marsh. Lim-
nology and Oceanography 24 (1979) 999 – 1013.
Huber, M., Meyer, U. and Rys, P.: Biodegradation mechanisms of linear alchol
ethoxylates under anaerobic conditions. Environ. Sci. Technol. 34 (2000)
1737 – 41.
ISO 11734: Water quality – Evaluation of the ultimate anaerobic biodegradability of
organic compounds in digested sludge – Method by measurement of the
biogas production (1995).
ISO/DIS 14 853: Evaluation of ultimate anaerobic biodegradation of plastic materi-
als in an aqueous system – Method by analysis of carbon conversion to carbon
dioxide and methane (1999).
ISWA (International Solid Waste Association) Handling, treatment and disposal of of
sludge in Europe. Situation Report 1 ISWA Working Group on Sewage sludge
and and Water Works, 94 pages, Copenhagen, 1995.
Jensen, J.: Fate and effects of linear alkylbenzene sulphonates (LAS) in the terres-
trial environment – a review. Sci. Tot. Environ. 226 (2 – 3) (1999) 93 – 111.
Jensen, J., Løkke, H., Holmstrup, M., Krogh, P. H. and Elsgaard, L.: Effects and risk
assessment of linear alkylbenzene sulfonates in agricultural soil. 5. Probabilistic
risk assessment of linear alkylbenzene sulfonates in sludge-amended soils.
Environ. Toxicol. Chem. 20 (8) (2001) 1690 – 1697.
Jensen, J. and Jepsen, S. E.: The production, use and quality of sewage sludge in
Denmark. Waste Management 25 (3) (2005) 239 – 247.
Jensen, J., Smith, S. R., Krogh, P. H., Versteeg, D. J. and Temara, A.: Risk assessment
of LAS in agricultual soils revisited II: species sensitivity distribution and risk
quotients. Chemosphere, in press (2007).
Jobst, H.: Chlorphenole und Nonylphenole in Klärschlämmen Teil 1: Vorkommen in
Klärschlämmen westdeutscher Kläranlagen aus den jahren 1987 bis 1989.
Acta hydrochim hydrobiol. 23 (1995) 20 – 25.
Jones, J. G.: Activities of aerobic and anaerobic bacteria in lake sediments and their
effect on the water column. In Sediment Microbiology, eds. D. B. Nedwell and
C. M. Brown, 107–145. Academic Press: London, 1982.
Jørgensen, B. B.: Bacterial sulfate reduction within reduced microniches of oxidised
marine sediments. Marine Biology 41 (1977a) 7–17.
Jørgensen, B. B.: The sulfur cycle of a coastal marine sediment (Limfjorden, Den-
mark). Limnology and Oceanography 22 (1977b) 814 – 832.
Jørgensen, B. B.: Mineralization of organic matter in the sea bed – the role of sulfate
reduction. Nature 296 (1982) 643 – 645.
Kaspar, H. F. and Tiedje, J. M.: Anaerobic bacteria and processes. In Methods of Soil
Analysis, Part 2. Chemical and Microbiological Properties, 989 – 1009. ASA-
SSSA: Madison, 1982.
Kayser, R.: Upgrading the wastewater treatment plant of the city of Oldenburg. Wat.
Sci. Tech. 29 (1994) 89 – 95.
Klein, Megauhey: The fate of detergents in septic tank systems and oxidation
ponds, Univ. of California, Report No. 64 (1), Jan. (1964).
Kloepper-Sams, P., Torfs, F., Feijtel, T. and Gooch, J.: Effects assessments for surfac-
tants in sludge-amended soils: a literature review and perspectives for TRA,
The Science of the Tot. Environment, 185 (1996) 171 – 185.
Klotz, H.: Alcohol ethoxylates (AE) in sewage treatment sludges. Interim results of a
German ring test. Analytica Conference 98, April 21 – 24, München.
Krogh, P. H.: Holmstrup and Jensen J. (1997). NERI report N. 69 (1998) 16.
Krogh, K. A., Vejrup, K. V., Mogensen, B. and Halling-Sorensen, B.: Liquid chroma-
tography mass spectrometry method to determine alcohol ethoxylates and al-
kylamine ethoxylates in soil interstitial water, ground water and surface water
samples. Journal Chromatogr., A. 957 (2002) 45 – 57.
Kroiss, H., Schweighofer, P., Frey, W. and Matsché, N.: Nitrification inhibition – a
source identification method for combined municipal and/or industrial waste-
water treatment plants. Wat. Sci. Tech. 26 (1992) 1135 – 1146.
Küchler, T.: Behaviour of surfactants and their influence on the mobility of organic
micropollutants in sandy soils with weak sorption capacities, Fraunhofer Insti-
tute, Thesis, Potsdam, 1995.
Kuhn, E. P., Zeyer, J., Eicher, P. and Schwarzenbach, R. P.: Anaerobic degradation of
alkylated benzenes in denitrifying aquifer columns. Appl. Environ. Microbiol. 54
(1988) 490 – 496.
Kujawa, M., Schnaak, W. and Kuechler, T.: Occurrence of organic pollutants in sew-
age sludge and influence of surfactants on their mobility in amended soils.
Fraunhofer Internal Report (1996).
Kunkel, E.: Umweltanalytik von Tensiden. Tenside Surf. Det. 24 (1987) 280 – 285.
Küster, E. and Niese, G.: Dumping of refuse and sludges. In Biotechnology Vol. 8.
Microbial Degradations, ed. W. Schönborn, VCH, (1986) 349 – 362.
Lara Martín, P. A., Gómez Parra, A. and González Mazo, E.: Pressurized liquid ex-
traction followed by liquid chromatography-mass spectrometry for the deter-
mination of mayor surfactants in marine sediments. International Journal of
Environmental Analytical Chemistry, Vol. 85 (4 – 5) April (2005) 293 – 303.
Lara-Martin, P. A., Gomez-Parra, A. and Gonzalez-Mazo, E.: Simultaneous extraction
and determination of anionic surfactants in waters and sediments. Journal of
Chromatography A , 1114 (2) (2006) 205 – 210.
Lara-Martin, P. A., Petrovic, M., Gomez-Parra, A., Barcelo, D. and González Mazo, E.:
Presence of surfactants and their degradation intermediates in sediment cores
and grabs from the Cadiz Bay area. Environmental Pollution 144 (2006) 483 –
491.
Lara-Martin, P. A., Gomez-Parra, A., Köchling, T., Sanz, J. L., Amils, R. and Gonzalez-
Mazo, E.: Anaerobic degradation of linear alkylbenzene sulfonates in coastal
marine sediments, Environmental Science & Technology, 41 (10) (2007)
3573 – 3579.
341
J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review
Lara-Martin, P. A., Gomez-Parra, A., Köchling, T., Sanz, J. L., Amils, R. and Gonzalez-
Mazo, E.: Monitoring the primary biodegradation of linear alkylbenzene sulfo-
nates and their coproducts in anoxic sediments using liquid chromatography-
mass spectrometry, Environmental Science & Technology, 41 (10) (2007)
3580 – 3586.
Larson, R. J., Federle, T. W., Shimp, R. J. and Ventullo, R. M.: Behaviour of LAS in soil
infiltration and ground water, Tenside Surf. Det. 26 (2) (1989) 116.
Latham, M. J.: The animal as an environment. In Microbial Ecology: A Conceptual
Approach, eds. J. M. Lynch and N. J. Poole, 115 – 137. Blackwell Scientific Pub-
lications: Oxford, 1979.
Laue, H., Denger, K. and Cook, A. M.: Taurine reduction in anaerobic respiration of
Bilophila wadsworthia RZATAU, Appl. Environ. Microbiol., in press (1997).
Lee, H. B. and Peart, T. E.: Determination of 4-nonylphenol in effluents and sludge
from sewage treatment plants, 67 (1995) 1976 – 1980.
León, V. M., Gonzalez-Mazo, E., Forja, J. M. and Gomez-Parra, A.: Identification of
linear alkylbenzene sulfonate biodegradation intermediates in anoxic marine
coastal sediments. Proceedings of the 5th World Surfactant Congress. Comite
Européen des Agents de Surface et Leurs Intermédiaires Organiques. Milano,
Italy, 2000.
León, V., González Mazo, E., Forja Pajares, J. M. and Gómez Parra, A.: Vertical dis-
tribution profiles of Linear Alkylbenzene Sulfonates and their long-chain inter-
mediate degradation products in coastal marine sediments. Environmental
Toxicology and Chemistry, Vol. 20 (10) (2001) 2171 – 2178.
Lie, T. J., Pitta, T., Leadbetter, E. R., Godchaux III, W. and Leadbetter, J. R.: Sulfonates:
novel electron acceptors in anaerobic respiration, Arch Microbiol., 166 (1996)
204 – 210.
López Serrano, I., Ferrer, J., Cassani, G., Ptavettoni, S. and Varo, P.: Study of LAS and
soap in marine sediments. Proceedings of the 32th annual CED meeting,
(2001) 51 – 65.
Lowe, S. E., Jain, M. K. and Zeikus, J. G.: Biology, ecology, and biotechnological ap-
plications of anaerobic bacteria adapted to environmental stresses in tem-
perature, pH, salinity, or substrates. Microbiological Reviews 57 (1993) 451 –
509.
Madsen, T., Boyd, H. B., Nylén, D., Petersen, A. R., Petersen, G. I. and Flemming S.:
Environmental and Health Assessment of Substances in Household Deter-
gents and Cosmetic Detergent Products. Environmental Project No. 615, Mil-
joeprojekt, CETOX (2001).
Madsen, T. and Rasmussen, H. B.: Evaluation of a method for determining the
anaerobic biodegradability of surfactants. Report No. 38 VKI Water Quality In-
stitute, Horsholm, Denmark, 1994.
Madsen, T., Rasmussen, H. B. and Nilsson, L.: Anaerobic Biodegradation Potentials
in Digested Sludge, Chemosphere 31 (1995) 4243 – 4258.
Madsen, T., Petersen, G., Seiero, C. and Torslov, J.: Biodegradability and Aquatic
Toxicity of Glycoside Surfactants and a Nonionic Alcohol Ethoxylate; JAOCS, 73
(1996) 929 – 933.
Madsen, T., Kristensen, P., Samsoe-Petersen, L., Torslov, J. and Rasmussen, J. O.:
Application of sludge on farmland – quality objectives, level of contamination
and environmental risk assessment. Proc. Specialty conference on manage-
ment and fate of toxic organics in sludge applied to land. Copenhagen, Den-
mark, 30 April – 2 May 1997.
Mara, D. D. and Pearson, H.: Artificial freshwater environment: Waste stabilisation
ponds. In Biotechnology Vol. 8. Microbial Degradations, ed. W. Schönborn,
(1986) 177 – 206.
Marcomini, A. and Giger, W.: Simultaneous determination of linear alkylbenzene
sulfonates, alkylphenol polyethoxylates, and nonylphenol by high performance
liquid chromatography. Anal. Chem. 59 (1987) 1709 – 1715.
Marcomini, A., Cape, P. D., Lichtensteiger, Th., Brunner, P. H. and Giger, W.: Behav-
iour of aromatic surfactants and PCBs in sludge- treated soil and landfills, J.
Environ. Qual. 18 (1989) 523 – 528.
Marin, M., Pivotti, L., Campesan, G., Turchettto, M. and Tallandini, L.: Effects and fate
of sediment sorbed linear alkylbenzene sulfonate (LAS) on the bivalve mollusc
Mitilus galloprovincialis Lmk. Water Research 28 (1994) 85 – 90.
Matsché, N.: Kombinierter Einsatz von biologischer und chemischer P-Elimination.
Gewässerschutz, Wasser, Abwasser 98 (1987) 13 – 33.
McAvoy, D. C., Eckhoff, W. S. and Rapaport, R. A.: Fate of Linear Alkylbenzene Sul-
fonate in the environment. Environ. Tox. and Chem. 12 (1993) 977 – 987.
McAvoy, D. C., White, C. E., Moore, B. L. and Rapaport, R. A.: Chemical fate and
transport in a domestic septic system: sorption and transport of anionic and
cationic surfactants. Env. Tox. Chem. 13 (1994) 213 – 221.
McEvoy, J. and Giger, W.: Accumulation of linear alkyl benzene sulfonates in sewage
sludges. Naturwissenschaften 72 (1985) 429 – 431.
Matthijs, E. and De Henau, H.: Determination of LAS in aqueous samples, sedi-
ments sludges and soils using HPLC. Tenside Surf. Det. 24 (1987) 4 (1987)
193 – 199.
Matthijs, E., Debaere, G., Itrich, N., Masscheleyn, P., Rottiers, A. and Stalmans, M.:
TW Federle, The fate of detergent surfactants in sewer systems, Wat. Sci.
Tech. 31 (1995) 321 – 328.
Matthijs, E., Burford, M. D., Cassani, G., Comber, M., Eadsforth, C., Haas, P.,
Klotz, H., Spilker, R., Waldhoff, H. and Wingen, H.-P.: Determination of Alcohol
Ethoxylate Components in Sewage Sludge. Tenside Surf. Det. 41 (3) (2004)
113 – 120.
Metzner, G. and Lemmer, H.: Semi-Continuous Tests for Simulation of Municipal
Sludge Digestion to Evaluate Anaerobic Processes. International Symposium
Environmental Biotechnology, Part II. Publisher: Technological 3 Institute, Ost-
ende, 1997.
Metzner, G.: Elimination von Dodecylbenzolsulfonsäure und Methylglycindiessig-
säure in anaerobem Milieu. Tenside Surf. Det. 35 (1) (1998) 46 – 51.
Metzner, G.: Ausgewählte aerobe und anaerobe Abbautestverfahren für Tenside.
Münchener Beiträge zur Abwasser-, Fischerei- und Flussbiologie 54 (2001)
263.
Mieure, J. P., Waters, J., Holt, M. S. and Matthijs, E.: Terrestrial safety assessment of
linear alkylbenzene sulfonate. Chemosphere 21 (1 – 2) (1990) 251 – 262.
342
Miranda, L. A. S., Henriques, J. A. P. and Monteggia, L. O.: Performance of UASb
and DAEB reactors in the anaerobic digestion of synthetic wastewater con-
taining sodium oleate and sodium stereate. Water Science & Technology
Vol. 54 (2) (2006) 127 – 133.
Mix-Spagl, K.: Untersuchungen zum Umweltverhalten von Seifen. In: Umweltverträg-
lichkeit von Wasch- und Reinigungsmitteln (Münchener Beiträge zur Abwasser-,
Fischerei- und Flussbiologie, Vol. 44) R. Oldenburg Verlag, München, 1990.
Mogensen, A. S. and Ahring, B. K.: Formation of metabolites during biodegradation
of linear alkylbenzene sulfonate in an upflow anaerobic sludge bed reactor
under thermophilic conditions. Biotechnol. Bioengineer. 77 (2002) 483 – 488.
Mogensen, A. S., Haagen, F. and Ahring, B. K.: Anaerobic degradation of Linear Al-
kylbenzene Sulfonate. Environmental Toxycology and Chemistry, Vol. 22 (4)
(2003) 706 – 711.
Mogensen, A. S., Hagensen, F. and Ahring, B. K.: Anaerobic biodegradation of linear
alkylbenzene sulfonate. Environ. Toxicol. Chem. 22 (2003) 706 – 711.
Moreno, A., Ferrer, J. and Berna, J. L.: Biodegradability of LAS in a sewer system,
Tenside Surf. Det. 27 (5) (1990) 312 – 315.
Moreno, A., Ferrer, J., Ruiz, B. F., Prats, D., Vazquez, B. and Zarzo, D.: Monitoring of
Linear alkylbenzene Sulfonate removal in a lagoon treatment system. CESIO
international surfactants congress, London, 1992.
Moreno, A. Bravo, J., Ferrer, J. and Bengoechea, C.: Soap determination in sewage
sludge by high-performance liquid chromatography. JAOCS 70 (1993) 667 –
671.
Morrall, S. W., Dunphy, J. C., Cano, M. L., Evans, A., McAvoy, D. C., Price, B. P. and
Eckhoff, W. S.: Removal and environmental exposure of alcohol ethoxylates in
US sewage treatment. Ecotoxicol. Environ. Saf.: 64 (2006) 3 – 13.
Mosey, F. E.: Anaerobic processes. In Ecological Aspects of Used-water Treatment.
Vol. 2. Biological Activities and Treatment Processes, eds. C. R. Curds and H. A.
Hawkes, 219 – 260. Academic Press, London, 1983.
Mosey, F. E.: Redox potentials in wastewater treatment. The Chemical Engineer,
May 1985, 21 – 24.
Mountfort, D. O., Asher, R. A., Mays, E. L. and Tiedje, J. M.: Carbon and electron flow
in mud and sandflat intertidal sediments at Delaware Inlet, Nelson, New Zeal-
and. Applied and Environmental Microbiology, 39 (1980) 686 – 694.
Müller, M. T.: Anaerobic Biodegradation and Toxicity of Alcohol Ethoxylates. Thesis,
ETH Zürich, 2000.
Nuck, B. A. and Federle, T. W.: A batch test for assessing the mineralisation of 14C-
radiolabeled compounds under realistic anaerobic conditions. Environmental
Science and Technology 30 (1996) 3597 – 3603.
OECD. OECD Guideline 308 for the Testing of Chemicals: Aerobic and Anaerobic
Transformation in Aquatic Sediment Systems. OECD, Paris, 2002.
OECD. Anaerobic Biodegradation of Organic Compounds in Digested Sludge –
Method by Measurement of Gas Production. Draft New Guideline 311 (Octo-
ber 2003 version). Organisation for Economic Cooperation and Development,
Paris, 2003.
Osburn, Q. W.: Analytical methodology for linear alkylbenzene sulfonate (LAS) in
waters and wastes. J. Am. Oil. Chem. Soc. 63 (1986) 257 – 263.
Pagga, U. and Beimborn, D. B.: Anaerobic Biodegradation Test for Organic Com-
pounds. Chemosphere 27 (1993) 1499.
Painter, H. A. and Zabel, T. F.: Review of the Environmental Safety of LAS. Report for
ECOSOL and SDIA, Water Research Centre, Medmenham, 1988.
Painter, H. A. and Zabel, T.: The behaviour of LAS in sewage treatment. Tenside Surf.
Det. 26 (2) (1989) 108 – 115.
Painter, H. and Mosey, F.: The question of the anaerobic biodegradability of LAS.
Proceedings of CESIO World Surfactant Congress, London (UK), 3 (1992) 34 –
43.
Painter, H. A.: Anionic surfactants. In The Handbook of Environmental Chemistry
Vol. 3 Part F, ed. O. Hutzinger, Springer-Verlag: Berlin, 1992.
Painter, H. A.: Detailed Review paper on Biodegradability Testing. OECD Paris, 1994.
Petersen, G. I. and Andersen, T. T.: DHI report no. 10 2004. DHI, Denmark 2004.
Petrovic, M., A., Fernandez-Alba, R., Borrull, F., Marce, R. M., Mazo, E. G. and Bar-
celò, D.: Occurrence and distribution of nonionic surfactants, their degradation
products, and linear alkylbenzene sulfonates in coastal, waters and sediments
in Spain. Environ. Toxycol. Chem. 2 (2002) 37 – 46.
Petzi, S.: Untersuchungen zum anaeroben Abbau von Seifen. Seifen-Öle-Fette-
Wachse 115 (1989) 229 – 232.
Pittinger, C. A. and Kimerle, R. A.: Abstract #455, p. 112. SETAC 11th Annual Meeting,
Alexandria, VA (1991).
Prats, D., Ruiz, F., Vasquez, B. and Rodriguez-Pastor, M.: Removal of anionic and
nonionic surfactants in a wastewater Treatment plant with anaerobic digestion.
A comparative study. Wat. Res. 31 (8) (1997) 1925 – 1930.
Prats, D., Rodriguez, M., Varo, P., Moreno, A., Ferrer, J. and Berna, J. L.: Biodegrada-
tion of soap in anaerobic digesters and on sludge amended soils. Wat.
Res. 33 (5) (1999) 1331 – 1336.
Prats, D., Rodriguez, M., Varo, P., Muela, M., Llamas, J. M., Moreno, A., Ferrer, J. and
Berna, J. L.: Elimination of xenobiotics during composting. A case study. LAS.
Tenside Surf. Det. 36 (5) (1999) 294 – 298.
Press, F. and Siever, R.: Rivers: Currents, channels, and networks. In Earth, (1986)
78 – 209.
Puchta, R., Krings, P. and Sandkühler, P.: A New Generation of Softeners. Tenside
Surf. Det. 30 (3) (1993) 186.
Randall, C. W., Barnard, J. M. and Stensel, H. D.: (Eds). Water Quality Management
Library Vol 5: Design and retrofit of wastewater treatment plants for biological
nutrient removal. Technomic Publishing Company, Lancaster, Pennsylvania,
1992.
Rapaport, R. A. and Eckhoff, W. S.: Monitoring linear alkyl benzene sulfonate in the
environment: 1973 – 1986. Environmental Toxicology and Chemistry 9 (1990)
1245 – 1257.
Rapaport, R. A., Larson, R. J., McAvoy, D. C., Nielson, A. M. and Trehy, M.: The fate of
commercial LAS in the environment. The CLER Review 1 (1995) 20 – 31.
Rehman, N., Melling, J., Van Duynhoven, J. and Roberts, D.: Fate of primary alkane
sulfates (PAS) under anaerobic conditions – Effect of braching on biodegrad-
ability. Poster presented at SETAC Europe, Lille, 2005.
Tenside Surf. Det. 44 (2007) 6
J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review

Reiser, R., Toljander, H. and Giger, W.: Historic record of alkylbenzenesulfonates in Swanwick, J. D., Shurben, D. G. and Jackson, S.: A survey of the performance of
recent lake sediments: input changes and postburial fate of detergent derived sewage sludge digesters in Great Britain. Journal of the Institute of Water Pol-
chemicals, Organic Geochemistry meeting, S. Sebastian, 1995. lution Control 6 (1969) 639 – 661.
Richards, F. R.: The Cariaco basin (Trench). Oceanography and Marine Biology An- Sweetman, A. J.: Development and application of a multi-residue analytical method
nual Review 13 (1975) 11 – 67. for the determination of n-alkanes, linear alkylbenzenes, polynuclear aromatic
Richards, B. N.: The Microbiology of Terrestrial Ecosystems. Longman Scientific and hydrocarbons and 4-nonylphenol in digested sewage sludges. Wat. Res. 2!
Technical: Harlow, 1987. (1994) 343 – 353.
Rottiers, A., Boeije, G., Corstanje, R., Decraene, K., Feijtel, T., Matthijs, E. and Scho- Tabor, C. F. and Barber, L. B.: Fate of linear alkylbenzene sulfonate in the Mississippi
wanek, D.: Adaptation of the CAS test system and synthetic sewage for bio- river. Environ. Sci. Technol. 30 (1996) 161–171.
logical nutrient removal. Accepted for publication in Chemosphere (1998). Takada, H. and Ishiwatari, R.: Linear alkylbenzenes in urban riverine environments in
Rudd, J. W. M. and Hamilton, R. D.: Methane cycling in Lake 227 in perspective with Tokyo: distribution, source, and behaviour. Environ. Sci Technol. 21 (1987)
some components of the carbon and oxygen cycles. Archiv für Hydrobiologie 875 – 883.
Ergebnisse der Limnologie 12 (1979) 115 – 122. Takada, H., Ishiwatari, R. and Ogura, N.: Distribution of linear alkylbenzenes (LABs)
Saez, M., Leon, V. M., Rubio, J. A., Gomez-Parra, A. and Gonzalez-Mazo, E.: A com- and linear alkylbenzene sulfonates (LAS) in Tokyo Bay sediments. Estuarine,
parative study of the adsorption of linear alkylbenzene sulfonates and their Coastal and Shelf Science 35 (1992) 141–156.
biodegradation intermediates on marine sediments and organisms. Ciencias Temper, U., Pfeiffer, W. and Bischofsberger, W.: Stand und Entwicklungspotentiale
Marinas 29 (4) (2003) 397 – 403. der anaeroben Abwasserreinigung unter besonderer Berücksichtigung der Ver-
Salanitro, J. P., Wisneiwski, H. L., Byers, D. L., Neaville, C. C. and Schroeder, R. A.: hältnisse in der Bundesrepublik Deutschland. Berichte aus Wassergüte-
Use of aerobic and anaerobic microcosms to assess BTEX biodegradation in wirtschaft und Gesundheitsingenieurwesen, Nr. 67, Technische Universität
aquifers. Ground water monitoring & Remediation 17 (3) (1997) 210 – 221. München, 1986.
Salanitro, J. P. and Diaz, L. A.: Anaerobic biodegradability testing of surfactants. Tschui, M. and Brunner, P. H.: Die Bildung von 4-Nonylphenol aus 4-Nonylphenol-
Chemosphere 30 (1995) 813 – 830. mono- und diethoxylat bei der Schlammfaulung, Vom Wasser, 65 (1985) 9 –
Sanderson, H., Price, B. B., Dyer, S. D., DeCarvalho, A. J., Robaugh, D., Waite, S. W., 19.
Morrall, S. W., Nielsen, A. M., Cano, M. L. and Evans, K. A.: Occurrence and US-EPA. Design manual Phosphorus Removal. EPA Technology Transfer Report EPA
hazard screening of alkyl sulfates and alkyl ethoxysulfates in river sediments. 625/1-87/001, Center for Environmental Research Information, Cincinnati,
Science of The Total Environment, 367 (1) (2006) 312 – 323. OH 45268 (1987).
Sanz, J. L., Rodriguez, M., Amils, R., Berna, J. L., Ferrer, J. and Moreno, A.: Anaerobic Vandepitte, V. and Debaere, G.: Biodegradation (aerobic/anaerobic) of dimethyl
biodegradation of LAS (Linear Alkylbenzene Sulfonate). Inhibition of the dodecyl amine oxide. Procter & Gamble ETC, Poster SETAC Congress San
methanogenic process. La Rivista Italiana delle sostanze grasse, Vol LXXVI, Lu- Francisco, 1997.
glio/Agosto, 1999. Varo Galván, P., Prats Rico, D. and Rodriguez Pastor, M.: Anaerobic biodegradation
Sanz, J. L., Culubret, E., de Ferrer, J., Moreno, A. and Berna, J. L.: Anaerobic biode- of soaps in anaerobic screening test. Tenside Surf. Det. 39 (5) (2002) 160 –
gradation of linear alkylbenzene sulfonate (LAS) in upflow anaerobic sludge 163.
blanket (UASB) reactors. Biodegradation 14 (2003) 57 – 64. Verge, C., Moreno, A. and Berna, J. L.: (1997). Influence of water hardness on the
Sedlak, R. I. and Booman, K. A.: A Study of LAS and alcohol ethoxylate removal at a bioavailability and toxicity of LAS. Chemosphere 44 (2001) 8.
municipal wastewater treatment plant. US SDA annual Convention, Boca Ra- Wagener, S. and Schink, B.: Anaerobic degradation of nonionic and anionic surfac-
ton, Fla (1986). tants in enrichment cultures and fixed-bed reactors, Wat. Res. 21 (1987) 615 –
Sieburth, J. M.: Contrary habitats for redox-specific processes: Methanogenesis in 622.
oxic waters and oxidation in anoxic waters. In Microbes in the Sea, ed. M. A. Wahlberg, C., Renberg, L. and Wideqvist, U.: Determination of nonylphenol and
Sleigh, 11 – 38. Ellis Horwood: Chichester, 1987. nonylphenol ethoxylates as their pentafluorobenzoates in water, sewage
Siegfried, M., Müller, M. T. and Baumann, U.: Anaerobic Degradation and Toxicity of sludge and biota. Chemosphere 20 (1990) 179 – 195.
Alcohol Ethoxylates in Anaerobic Screening Test Systems; CESIO Congress Waters, J. and Feijtel, T. C. J.: AISE/CESIO Environmental surfactant monitoring pro-
Barcelona, 3 (1996) 61 – 275. gramme; outcome of five national pilot studies on linear alkylbenzene sulfo-
Shimp, R. J., Lapsins, E. V. and Ventullo, R. M.: Chemical fate and transport in do- nate (LAS). Chemosphere 30 (1995) 1939 – 1956.
mestic septic system : biodegradation of Linear Alkylbenzene Sulfonate (LAS) Zehnder, A. J. B. and Stumm, W.: Geochemistry and biogeochemistry of anaerobic
and Nitrilotriacetic Acid (NTA). Environmental Toxicology and Chemistry, 13 (2) habitats. In ‘Biology of Anaerobic Microorganisms’, ed. A. J. B. Zehnder, 1 – 35,
(1994) 205 – 212. Wiley-Liss. Ahel, M., Giger, W. (1985). Determination of alkylphenols and alkyl-
Schlegel, H. G. and Jannasch, H. W.: Prokaryotes and their habitats. In The Prokar- phenol mono-and diethoxylates in environmental samples by high perform-
yotes: A Handbook on Habitats, Isolation and Identification of Bacteria, Vol. 1, ance liquid chromatography. Anal. Chem. 57 (1988) 1577 – 1583.
eds. M. D. Starr et al., 43 – 82. Springer-Verlag: New York, 1981.
Schöberl, P., Klotz, H. and Spilker, R.: (1996a). Alkylbenzolsulfonat (LAS)-Monitoring.
Teil 3: LAS-Gehalte der Schwebstoffe verschiedener deutscher Fliessgewässer.
Tenside Surf. Det. 33 (4) (1996) 329 – 335. Glossary of Abbreviations
Schöberl, P. and Spilker, R.: (1996b). Alkylbenzolsulfonat (LAS)-Konzentrationen im
Lippe-Sediment eines Rhein-Altarmes. Tenside Surf. Det. 33 (5) (1996) 400 – 403.
Schowanek, D., Feijtel, T., Rottiers, A., Matthijs, E., Decraene, K., Boeije, G. and Ver- y AE : Alcohol Ethoxylates
straete, W.: Development and validation of new continuous activated sludge y AES : Alcohol Ether Sulfates
test systems with biological nutrient removal. Proc. IAWQ-NVA Conference
Advanced Wastewater Treatment, Amsterdam, September 23 – 25 (1996) y AISE : Association International de la Savonnerie,
431 – 433. de la D
tergence et des Produits d’Entretien
Schowanek, D., David, H., Francaviglia R., Hall, J., Kirchmann, H., Krogh, H. P., y AOS : Alpha Olefin Sulfonates
Schraepen, N., Smith, S. and Wildemann, T.: Probabilistic Risk Assessment for
Linear Alkylbenzene Sulfonate (LAS) in Sewage Sludge used on Agricultural y APG : Alkylpoliglucosides
Soil. Regulatory Toxicology and Pharmacology 49 (2007) 245 – 259. y APEO : Alkyl Phenol Ethoxylates
Shelton, D. R. and Tiedje, J. M.: General Method for Determining Anaerobic Biode-
gradation Potential. Appl. Env. Microbiol. 47 (1984) 850 – 857. y AS : Alcohol Sulfates, Alkyl Sulfates
Shokes, R. F., Trabant, P. K., Presley, B. J. and Reid, D. F.: Anoxic, hypersaline basin in y BABS : Branched Alkylbenzene Sulfonate
the northern Gulf of Mexico. Science 196 (1977) 1443 – 1446. y CEFIC : European Federation of Chemical Industries
Smolenski, W. J. and Suflita, J. M.: Biodegradation of cresol isomers in anoxic aqui-
fers. Applied and Environmental Microbiology 53 (1987) 710 – 716. y CESIO : Comit
Europ
en des Agents de Surface et
Steber, J. and Wierich, P.: The Anaerobic Degradation of Detergent Range Fatty Al- leurs Interm
diaires Organiques
cohol Ethoxylates. Studies with 14C-Labelled Model Surfactants, Water Res. 21 y DIC : Dissolved Inorganic Carbon
(1987) 661 – 667.
Steber, J., Gode, P. and Guhl, W.: The ecological evaluation of a group of important y DM : Dry Matter
detergent surfactants – fatty alcohol sulfates. Soap/Cosmetics/Chemical Spe- y DTDMAC : Di Tallow Di Methyl Ammonium Chloride
cialities 64 (1988) 44 – 50.
Steber, J. and Wierich, P.: The environmental fate of fatty acid -sulfomethyl esters:
y ECETOC : European Centre of Ecotoxicology and Toxi-
biodegradation studies with a carbon – 14 labelled model surfactant. Tenside, cology of Chemicals
26 (1989) 406 – 411. y EEA : European Environment Agency
Steber, J.: Wie vollständig sind Tenside abbaubar?, Textilveredelung 26 (1991) 348 –
354. y EPA : Environmental Protection Agency
Steber, J. and Birch, R. R.: Chemical structures and their biodegradability in the y ERASM : Environmental Risk Assessment and Man-
anaerobic environment. Final Report of EU Project Contract No. STEP-CT 91- agement
0154 (1995a).
Steber, J., Herold, C.-P. and Limia, J. M.: Comparative evaluation of anaerobic bio- y HAD : Haupt-Ausschuss Detergentien
degradability of hydrocarbons and fatty derivatives currently used as drilling y HERA : Human and Environmental Risk Assess-
fluids. Chemosphere 31 (1995b) 3105 – 3118.
Steber, J., Guhl, W., Stelter, N. and Schröder, F. R.: (1995c). Alkyl polyglycosides –
ment
ecological evaluation of a new generation of nonionic surfactants, Tenside Surf. y HPLC : High Performance Liquid Chromatography
Det. 32 (6) (1995) 515 – 521. y ILSI : International Life Sciences Institute
Strotmann, U. J. and Eglsäer, H.: The toxicity of substituted phenols in the nitrifica-
tion inhibition test and luminescent bacteria test. Ecotoxicol. Environ. Saf., y ISO : International Standarisation Office
30 (3) (1995) 269 – 273. y LAS : Linear Alkylbenzene Sulfonates

Tenside Surf. Det. 44 (2007) 6 343

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review

y LeAF : Lettinga Associates Foundation y TMAC : Tri Methyl Alkyl ammonium Chloride
y MES : Methyl Esther Sulfonates y UASB : Up Flow Anaerobic Sludge Blanket reactor
y OECD : Organisation of Economic Cooperation and
Development
y SAS : Secondary Alkane Sulfonates Appendix
y SCHER : Scientific Committee on Health and Envir-
onmental Risks Overview of surfactant monitoring and mass balancing in
y SPC : Sulpho Phenyl Carboxilates anaerobic environmental compartments
y STP : Sewage Treatment Plant

Name Surfac- Type Monitoring Concentration Environ- Removal Resi- Remarks References
tant site levels mental results dence
(mg/kg DM) conditions (%) Time
Anionics
Sulfonates LAS Commercial Digestor 2900 – 11900 Anaerobic M McEvoy & Giger
sludges (1985)
LAS Commercial Digestor 4200 (1200) Anaerobic 20 – 29 digestors/grab Giger et al., (1987)
sludges 30 % samples
LAS Commercial Digestor 9300 – 18 800 Anaerobic M Grab samples; one Holt & Bernstein
sludges digestor (1992)
LAS Commercial Digestor 6660 Anaerobic 0% Sedlak et al.,
sludges (1986)
LAS Commercial Digestor 4660 (1540) Anaerobic M US five plants – Rapaport & Eckhoff
sludges 49 grabs (1990)
LAS Commercial Digestor 6000 – 9400 Anaerobic M Monitoring in Spain Waters & Feijtel
sludges & Italy (1995)
LAS Commercial River 100 – 322 undefined M Osburn (1986)
sediment
LAS Commercial river SS 2 – 209 not M Schöberl et al.,
applicable (1996)
LAS Commercial River 0 – 3.3 Undefined M Monitoring cores of Schöberl & Spilker
sediment Lippe sediment (1996)
from 1939 – 1991
LAS Commercial Anaer. pond 5.2 – 6.3 mg/l Undefined ca. 20 % 20 to Over a period of Moreno et al.,
water Eh 60 days one year (1994)
LAS Anaer. pond 43 600 Moreno et al.,
sedim. (1994)
LAS Tridecyl- Fresh water 0.48 (spiked) Anaerobic 0% Lab study with Federle & Schwab
benzene pond pond sediment in- (1992)
oculum
LAS Commercial River 0.49 – 5.3 undefined M Monitoring in Spain Waters & Feijtel
sediment & Italy (1995)
LAS Commercial River 0.01 – 20 undefined M Monitoring in Mis- Tabor & Barber
sediment sisipi (1996)
LAS Commercial River 0.6 – 567 undefined M Monitoring in Tokyo Takada & Ishiwatari
sediment area (1987)
LAS Commercial River 5.6 undefined M Marcomini & Giger
sediment (1987)
LAS Commercial Marine 10 to 30 Undefined M Matthijs et al.,
sediment Eh (1986)
LAS Commercial Marine 11 to 30 Undefined M Hon-Nami & Hanya
sediment Eh (1980)
LAS Commercial Landfill 245 (bottom) Undefined M Landfill receiving Marcomini et al.,
– 9160 (top) Eh digested sludge (1989)
LAS Commercial River 0.5 to M Monitoring of Bot- Cavalli et al.,
sediment 4.74 mg/kg tom sediments in (2000)
Po river (Italy)
20 points
LAS Commercial Marine 1.2 – 26.7 mg/ M 5 points in Cadiz DelValls et al.,
sediment kg in summer bay and 2 points in (2002)
1.2 – 62.1 in Barbate river
winter

344 Tenside Surf. Det. 44 (2007) 6

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review

Name Surfac- Type Monitoring Concentration Environ- Removal Resi- Remarks References
tant site levels mental results dence
(mg/kg DM) conditions (%) Time
Sulfonates LAS Commercial Marine 0.5 – 1 mg/kg M Branched sulfo- Folke et al., (2003)
sediment in coast nates were also
2 – 20 mg/kg in detected in some
port samples
LAS Commercial Marine 138 mg/kg SPC in the range Leon V. M. et al.,
sediment close to 0.9 – 0.07 mg/kg (2001)
discharge were also detected
0.8 – 16.4 in
other points
LAS Commercial Marine sed 0.039 – M Several locations in Bester et. al.,
in Elbe 0.109 mg/kg the Elbe estury at (2001)
estuary 5 m depth
LAS Commercial Sludge < 1 mg /kg 8 field sites in the Carlsen (2002)
amende 20 mg/kg in North of Spain
soil heavy
amended soil
for 25 years
LAS Commercial Marine sed 0.538 – M Guadalete estuary Lara-Martin et al.,
in estuary 1.014 mg/kg (Es) (2006)
LAS Commercial Sludge 0.12 – M LAS and SPC in 10 Eichhorn, Peter
amended 12.84 mg/kg field sites, 10days et al., (2005)
soil for LAS to 4 years after
0.004 – 10.220 sludge treatment
for SPC
SAS Commercial Digestor 648 – 738 Anaerobic 0% 40 days 10 day sampling Field et al., (1995)
sludges
Sulfates AES Septic Anaerobic 72 – 81 % No environment Birch et al., (1992)
Tanks characterisation
AS Digestor 66 % to be completed
Soaps Soap Digestor 18 000 – 51900 Anaerobic 70 – 75 % 26 days Sampling over sev- Moreno et al.,
sludges eral treatment (1993)
plants
Nonionics
Alcohol AE Commercial Digestor 2600 (400) Anaerobic 65 % 26 days One day sampling Prats et al., (1997)
Ethoxylates
AE Commercial Digestor < 500 typically Anaerobic M Monitoring pro- Klotz (1998)
gramme in Ger-
many (Tegewa)
Alkylphe- NP Digestor 900 (600) Anaerobic M 29 digestors Giger et al., (1987)
nol Ethoxy- sludges
lates
NP Digestor 450 – 2530 Anaerobic M Several digestors Giger et al., (1984)
sludges
NP Digestor 1600 Anaerobic M Ahel & Giger
sludges (1985)
NP + Digestor 3 – 540 Anaerobic M Madsen et al.,
NPEO sludges (1997)
NP Digestor 35 – 95 Anaerobic M Kujawa et al.,
sludges (1996)
NP Digestor 545 – 1000 Anaerobic M Non Indicated Ahel et al., (1994)
sludges
NP Digestor 130 – 400 Anaerobic M Kunkel (1987)
sludges
NP Digestor 640 – 2200 Anaerobic M Brunner et al.,
sludges (1988)
NP Digestor 137 – 470 Anaerobic M Lee & Peart (1995)
sludges
NP Digestor 1200 Anaerobic M Marcomini & Giger
sludges (1987)
NP Digestor 400 – 1200 Anaerobic M Wahlberg et al.,
sludges (1990)
NP Digestor 20 – 350 Anaerobic M Chalaux et al.,
sludges (1994)

Tenside Surf. Det. 44 (2007) 6 345

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review

Name Surfac- Type Monitoring Concentration Environ- Removal Resi- Remarks References
tant site levels mental results dence
(mg/kg DM) conditions (%) Time
Alkylphe- NP Digestor 21 – 1193 Anaerobic M Jobst (1995)
nol Ethoxy- sludges
lates
NP River 0.29 – 6.73 undefined M Lee & Peart (1995)
sediment
NP River
sediment
NP River sedi- 0.9 undefined M Marcomini & Giger
ment (1987)
NP River 0.02 – 0.04 undefined M Chalaux et al.,
sediment (1994)
NP Marine se- 0.006–0.07 undefined M Chalaux et al.,
diment (1994)
NP Subsurface 0.2 – 0.96 lg/l Mixed Eh/ 90 % 2–4 h Soil biodegradation Ahel et al., (1996)
(aquifier) (range 0.0 – Aerobic of NP
33)
NP1EO Digestor 5 – 40 Anaerobic M Kunkel (1987)
sludges
NP2EO Digestor <3 Anaerobic M Kunkel (1987)
sludges
NP1EO Digestor 90 – 680 Anaerobic M Brunner et al.,
sludges (1988)
NP2EO Digestor 20 – 220 Anaerobic M Brunner et al.,
sludges (1988)
NP1EO Digestor 220 Anaerobic M Marcomini & Giger
sludges (1987)
NP2EO Digestor 30 Anaerobic M Marcomini & Giger
sludges (1987)
NP1EO Digestor 20 – 190 Anaerobic M Wahlberg et al.,
sludges (1990)
NP2EO Digestor 1.0 – 50.0 Anaerobic M Wahlberg et al.,
sludges (1990)
NP1EO River 0.8 undefined M Marcomini & Giger
sediment (1987)
NP2EO River 0.7 undefined M Marcomini & Giger
sediment (1987)
NP1EO Subsurface 0.04 – 0.91 lg/l Mixed Eh/ 99 % 2–4 h Soil biodegradation Ahel et al., (1996)
(aquifier) (range 0.0 – Aerobic of NP
4.9)
NP2EO Subsurface 0.01 – 0.33 lg/l Mixed Eh/ 99 % 2–4 h Soil biodegradation Ahel et al., (1996)
(aquifier) (range 0.00 – Aerobic of NP
23)
NP1EC Subsurface 2.9 – 10.9 lg/l Mixed Eh/ 80 % 2–4 h Soil biodegradation Ahel et al., (1996)
(aquifier) (range 0.00 – Aerobic of NP
13.1)

Received: 07. 08. 2007 The authors of this paper

Revised: 02. 10. 2007
José Luis Berna Tejero got his degree in chemistry at the University of Zaragoza
y Correspondence to (Spain) in 1969 and master in chemistry and technology of petroleum at the Uni-
versidad Complutense of Madrid in 1971. He joined Petresa in 1971 as a research
chemist and he is presently Director of R&D of the Petresa group. He is the author
Dr. José Luis Berna and co-author of numerous publications and participations in international con-
Petresa Group gresses mainly in the field of surfactant raw materials development, environmental
Campo de Las Naciones chemistry and risk assessment of surfactants.
Avenida Partenon, 12 – 14
E-28042 Madrid Giorgio Cassani studied chemistry and obtained his PhD at the University of Milan,
Spain Italy. From 1976 to 1982 he worked at the research centre Instituto Donegani
E-mail: jbt@petresa.com (Montedison Group), first in the field of synthesis and identification of new “insect
pheromones” of agricultural interest and later in the field of isolation/-identification
of microbial fermentation products (biotechnology group). In 1992 he moved to
EniChem Augusta (now Sasol Italy S.p.A.) as Head of the Analytical Laboratory at
the research centre in Paderno Dugnano (Milano). His main experience now is in
the field of surfactant: analysis, environmental determination and biodegradation
studies. He is author/co-author of more than 40 publications.

346 Tenside Surf. Det. 44 (2007) 6

J. L. Berna et al.: Anaerobic biodegradation of surfactants – scientific review
Claus-Dierk Hager holds a German diploma in chemistry and obtained his PhD in
metal organic chemistry at the University of Dortmund in 1979. In 1979 he joined
Unilever as a process development manager in the edible fats and oils business. In
1984 he moved to Hüls AG, Surfactants Business Unit, where he was responsible
for the application of surfactants in detergents and cleaners. After the acquisition of
the Hüls Surfactants Business by CONDEA Chemie GmbH and finally in 2001 by
SASOL, Claus-D Hager is now working for SASOL Germany, where he is managing
the Product Safety and Regulatory Affairs Department as well as Market Research.
Naheed Rehman is an environmental team leader advising Unilever on all aspects
of environmental safety. She holds a PhD and conducts global environmental risk
assessments for Unilever’s ingredients. She is responsible for delivery of Unilever’s
Safety & Environmental Assurance Centre (SEAC’s) ecotoxicology work programme.
Dr. Rehman leads and co-ordinates R&D projects for safety approval with specialist
expertise in assessing the effect of chemical structure on the biodegradability of
Unilever’s ingredients (aerobic and anaerobic). She has published several reports
and scientific papers in peer reviewed journals.
Ignacio López Serrano obtained his MSc in Chemistry by the University of Granada
in 1997. He got a Master in Engineering and Environmental Management by the
Escuela de Organización Industrial in 1999. At the end of 1999 he joined Petresa
working within the R+D division. His area of research comprises of the study of
raw materials, surfactant derivatives and detergency as well as the environmental
behaviour of surfactants. Currently he is responsible for the Customer Service and
Environmental Laboratory of Petresa.
Diederik Schowanek is a Bio-Engineer by training and obtained his PhD in Environ-
mental Engineering in 1991 from Gent University (Belgium). He is currently princi-
pal scientist in the Product Safety and Regulatory Affairs Department of Procter &
Gamble’s Brussels Innovation Centre (Belgium). Dr. Schowanek is involved in the
planning and management of environmental (research) activities of P&G. He has
built a recognised expertise in domains like Risk Assessment of chemicals and Life
Cycle Assessment. Dr. Schowanek is also actively involved in professional organisa-
tions such as B-IWA (Belgian Committtee of the International Water Association),
EWA (European Water Association), and the Royal Flemish Engineering Society
Tenside Surf. Det. 44 (2007) 6
(KVIV- Section ‘Environmental Technology’). With P&G he is also involved in various
EU-sponsored environmental research projects. Dr. Schowanek has published over
40 scientific papers in international journals and handbooks.
Josef Steber holds a diploma in biology and obtained his PhD in microbiology/bio-
chemistry at the University of Munich in 1976. He joined the Department of Ecology
at Henkel in 1978. Until his retirement end 2006 he was in charge of this depart-
ment and has been member of numerous international working groups dealing
with the environmental safety assessment of chemicals.
Klaus Taeger was born in 1954. He studied microbiology at the University of Göttin-
gen and obtained his PhD in Wuppertal in 1986. Until 1988 his main subject was
the bioremediation of soil. Afterwards he has worked in the departments of Emis-
sion Control and Ecology as well as Product Safety at BASF AG. An important part of
his work is to investigate the biodegradability of detergent ingredients.
Thorsten Wind holds a German Diploma of Biology and an American Bachelor of
Science. He finished his PhD-work in 1997 at the Max-Planck-Institute of Terrestrial
Microbiology in Marburg/Germany. After two years in the field of international pro-
ject management in the diagnostic industries Thorsten Wind started 1999 as a lab
manager of the ecological laboratories at Henkel KGaA- Düsseldorf/Germany. He is
currently working as a manager of Environmental Product Safety Assessment in the
Dept. of Corporate SHE & Product Safety responsible for the detergent sub-division.
His special expertise is in exposure analysis, environmental monitoring and model-
ling and biodegradation.
You will find the article and additional material by enter-
ing the document number TS100351 on our website at
www.tsdjournal.com
347