Biodegradation (2017) 28:303–312
DOI 10.1007/s10532-017-9797-6
ORIGINAL PAPER
Anaerobic digestion of amine-oxide-based surfactants:
biodegradation kinetics and inhibitory effects
Francisco Rı́os . Manuela Lechuga . Alejandro Fernández-Arteaga .
Encarnación Jurado . Mercedes Fernández-Serrano
Received: 29 September 2016 / Accepted: 28 May 2017 / Published online: 31 May 2017
Ó Springer Science+Business Media Dordrecht 2017
Abstract Recently, anaerobic degradation has
become a prevalent alternative for the treatment of
wastewater and activated sludge. Consequently, the
anaerobic biodegradability of recalcitrant compounds
such as some surfactants require a thorough study to
avoid their presence in the environment. In this work,
the anaerobic biodegradation of amine-oxide-based
surfactants, which are toxic to several organisms, was
studied by measuring of the biogas production in
digested sludge. Three amine-oxide-based surfactants
with structural differences in their hydrophobic alkyl
chain were tested: Lauramine oxide (AO-R12), Myris-
tamine oxide (AO-R14) and Cocamidopropylamine
oxide (AO-cocoamido). Results show that AO-R12
Electronic supplementary material The online version of
this article (doi:10.1007/s10532-017-9797-6) contains supple-
mentary material, which is available to authorized users.
F. Rı́os  M. Lechuga (&)  A. Fernández-Arteaga 
E. Jurado  M. Fernández-Serrano
Chemical Engineering Department, University of
Granada, Campus Fuentenueva, 18071 Granada, Spain
e-mail: nlvillen@ugr.es
F. Rı́os
e-mail: rios@ugr.es
A. Fernández-Arteaga
e-mail: jandro@ugr.es
E. Jurado
e-mail: ejurado@ugr.es
M. Fernández-Serrano
e-mail: mferse@ugr.es
and AO-R14 inhibit biogas production, inhibition
percentages were around 90%. AO-cocoamido did
not cause inhibition and it was biodegraded until
reaching a percentage of 60.8%. Otherwise, we fitted
the production of biogas to two kinetic models, to a
pseudo first-order model and to a logistic model.
Production of biogas during the anaerobic biodegra-
dation of AO-cocoamido was pretty good adjusted to
the logistics model. Kinetic parameters were also
determined. This modelling is useful to predict their
behaviour in wastewater treatment plants and under
anaerobic conditions in the environment.
Keywords Amine-oxide-based surfactants 
Anaerobic biodegradation  Biodegradation kinetics 
Biogas production  Digested sludge  Inhibition
Introduction
Surfactants are present in cleaning and personal
hygiene products, and therefore they are regular
constituents of wastewaters and the corresponding
receiving freshwater and saltwater (Sharvelle et al.
2008; Baena-Nogueras et al. 2014). At present, the
activated sludge process is one of the most commonly
method for the treatment of sewage waters (Jenkins
and Wanner 2014). However, surfactants can cause
important undesirable effects in wastewater treatment
plants by foaming in the aeration tanks of activated
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sludge or the inhibition of the autochthons aerobic
microorganisms (Liwarska-Bizukojc and Bizukojc
2008; Dereszewska et al. 2015). In addition, surfac-
tants are strongly sorbed on aerobic sludge due to their
hydrophobic character (Baena-Nogueras et al. 2013).
The excessive production of sludge containing pollu-
tants, remains as a difficult drawback for the activated
sludge process (Martı́nez-Garcı́a et al. 2016).
Anaerobic digestion is one of the most common
post treatments of sludge in wastewater treatment
plants (Huang and Goel 2015). The use of anaerobic
digested sludge in agriculture as fertilizers is a
considerably potential source of surfactants in the
terrestrial environment (Soares et al. 2008; Zhang
et al. 2015; Richter et al. 2016). Therefore, the fate of
surfactants, and particularly their decomposition in
activated sludge processes, plays an important role in
minimizing environmental impact. Besides, the secu-
rity use of municipal sewage sludge has become an
issue of particular concern of the European Commis-
sion (WD 2000, 2010); several European states have
established limit values for organic pollutants, includ-
ing surfactants, in sewage sludge (Kapanen et al.
2013).
Recently, anaerobic digestion has become a preva-
lent alternative for the treatment of wastewater. It is
acknowledged as the most cost-effective way for
waste activated sludge treatment and the generation of
bioenergy resources (Zhou et al. 2015), has excellent
organic removal efficiency and requires small occu-
pation (Lettinga 2001). Moreover, anaerobic treat-
ment technologies is widely used as a source of
renewable energy by production of biogas rich in
methane (McCarty et al. 2011; Colmenar-Santos et al.
2016). The substitution of aerobic process by others
based on anaerobic technologies remains as a chal-
lenge nowadays (Lay et al. 2016; Martı́nez-Garcı́a
et al. 2016; Wang et al. 2016). Consequently, the
anaerobic biodegradability of recalcitrant substances,
such as surfactants, and the possible inhibition effects
to the methanogenic microorganisms require thorough
and detailed studies (Pedrazzani et al. 2012; Delforno
et al. 2016). Furthermore, a readily biodegradability is
one of the restrictive criteria established by the
European Union to obtain the ‘‘Eco-Label’’ in deter-
gent products (EC 66/2010).
Amine-oxide-based surfactants come under the
special class of surfactant known as amphoteric. They
are surfactants that turn cationic in acidic solutions,
123
Biodegradation (2017) 28:303–312
due to the protonation of the N–O amino group
(Baglioni et al. 2015). In the recent years, they are
increasingly used and their applications vary with the
length of the alkyl chain and the substituents of the
nitrogen atom of amine oxide (Singh et al. 2006).
Amine-oxide-based surfactants are excellent foam
boosters and foam stabiliser in blends based on other
anionic or amphoteric surfactants, they are extensively
used in high concentrated hand washing-up liquids,
detergents, antistatic preparations and their use is
recommended for a wide range of cosmetic and
household products due to its good tolerance by skin
and hair. Moreover, some studies reported about their
readily aerobic biodegradability (OECD SIDS 2006;
Jurado et al. 2013).
Other authors studied the aerobic and anaerobic
biodegradation of some amine-oxide-based surfac-
tants (Garcı́a et al. 2007). Nevertheless, the charac-
terization and modelling of the biodegradation profiles
need to be studied and kinetic parameters need to be
elucidated. Kinetic studies are important in under-
standing reactions, and they have practical implica-
tions in the anaerobic digestion in wastewater
treatment plants and during the biodegradation under
anaerobic conditions in the environment. This study
investigates the anaerobic biodegradation of three
amine-oxide-based surfactants with differences in
their hydrophobic chain by measuring of the biogas
production. We evaluated the inhibitory effects on the
anaerobic microorganisms. Moreover, we followed
the mass of the carbon produced as gas to draw the
biodegradation profiles and to obtain the anaerobic
kinetic model.
Materials and methods
Surfactants and reagents
Three amine-oxide-based surfactants were tested, AO-
R12, AO-R14, and AO-Cocoamido, provided by Kao
Corporation (Tokyo, Japan) under the commercial
name OXIDETÒDM-20, OXIDETÒDM-4 and
OXIDETÒL-75 C respectively. They are a particular
class of nitrogen containing amphoteric surfactants
that exhibit cationic behavior at pH below 7. They
have the same hydrophilic group, but they differ in
their hydrophobic chain (length and/or the presence of
an amide group). Figure 1 shows the structure of the
Biodegradation (2017) 28:303–312 305

Fig. 1 Structural formula

of amine-oxide-based
surfactants

amine-oxide based surfactants and Table 1 shows
their chemical identification and technical
specifications.
The other reagents used (KH2PO4, Na2HPO412H2-
O, NH4Cl, CaCl22H2O, MgCl26H2O, FeCl24H2O,
resazurin, Na2S9H2O, MnCl24H2O, H3BO3, ZnCl2,
CuCl2, Na2MoO42H2O, CoCl26H2O, NiCl26H2O
and Na2SeO3) were analytical grade and provided by
Panreac Quı́mica S.L.U (Barcelona, Spain). Solutions
were prepared using purified Milli-Q water. All
glassware was initially cleaned with a 16.4 g L-1
solution of (NH4)2S2O8 in H2SO4 (98%).
Critical micelle concentration
Surface tension measurements were carried out using
a tensiometer model K11 (Krüss GmbH, Hamburg,
Germany) equipped with a 2-cm platinum plate. The

Table 1 Chemical identification and technical specifications of amine-oxide-based surfactants

Surfactant Abbreviation R or R0 % Ca % Water contentb CMCc mg L-1 CMCc mM

Lauramine oxide AO-R12 12 22.01 70.7 160.6 0.685

Myristamine oxide AO-R14 14 22.41 72.0 107.7 0.134
Cocamidopropylamine oxide AO-Cocoamido R0 -CO: coconut 22.70 67.1 309.9 0.225
a
Carbon content: measurement of TOC (Total Organic Carbon) (Lechuga et al. 2014)
b
Determined using infrared radiation (Rı́os et al. 2016)
c
CMC critical micelle concentration measured at 25 °C

123
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plate was thoroughly cleaned and flame-dried in a
Bunsen burner before each measurement. The surface
tension was measured at different surfactant concen-
tration dipping the vertically plate into the solution
and then subsequently pulled out. Six successive
measurements were performed and the standard
deviation did not exceed ± 0.1 mN/m. The temper-
ature was controlled at 25 ± 0.1 °C by circulating
tempered water through the jacketed cell. The CMC
value was estimated from plots of surface tension as a
function of surfactant concentration in a semi-log plot.
Surface tension has a rapid linear decrease followed by
a slow decrease. The break point indicates the
formation of surfactant aggregation, defined as the
CMC. Table 1 summarizes the CMC values deter-
mined and Figs. S1, S2 and S3 in supplementary
material show the plots of surface tension versus
surfactant concentration for the three amine-oxide-
based surfactants tested.
Anaerobic biodegradation
The ultimate anaerobic biodegradation was evaluated
by measuring of biogas production as compared to a
control without surfactant. The assay was carried out
following the ISO 11734:1995 guideline (ISO
11734:1995). This method evaluates the extent of
ultimate anaerobic biodegradation of a chemical based
on production of biogas as compared to a blank
without the addition of the test substance. The
production of carbon dioxide and methane was
measured using the system Oxitop ControlÒ (WTW,
Welheim, Germany). This system consists of individ-
ual glass reactors (510 mL) equipped with a magnetic
stirred and sealed with a cap containing an electronic
pressure indicator which measures the pressure
increase in the reactors headspace. Much of the CO2
produced will be dissolved in the liquid phase or
transformed into carbonate or hydrogen carbonate
under the conditions of the test. This inorganic carbon
is measured at the end of the test using the TOC
analyser TOC-V CSH (Shimadzu, Kioto, Japan). The
amount of carbon (inorganic plus methane) resulting
from the anaerobic biodegradation of the test sub-
stance is calculated from the net gas production and
net IC formation in the liquid phase in excess of blank
control values.
Sludge sampled from the anaerobic digester of the
Granada WWTP (Granada, Spain, 37°090 54.100 N–
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Biodegradation (2017) 28:303–312
3°370 31.800 W) was used as inoculum. Sludge was
pre-incubated for seven days at 35 ± 1 °C to reduce
the non-specific production of gas. Table 2 shows the
characteristic parameters of the sludge determined
before use in the experiments following the Standard
Methods for the examination of water and wastewater
(APHA 1992). Immediately after their use, sludge was
centrifuged and washed with a mineral salt solution to
reduce the inorganic carbon (IC) to a value less than
10 mg L-1.
Amine-oxide-based surfactants were tested in trip-
licate at the initial concentration of 100 mg L-1
organic carbon in mineral medium. Organic carbon
concentration was measured using TOC analyser
TOC-V CSH (Shimadzu, Kioto, Japan). The mineral
salt solution was prepared adding in 1 L of de-
oxygenated water: 0.27 g KH2PO4, 1.12 g Na2HPO4-
12H2O, 0.53 g NH4Cl, 0.075 g CaCl22H2O, 0.10 g
MgCl26H2O, 0.02 g FeCl24H2O, 0.001 g resazurin,
0.1 Na2S9H2O and 10 mL of solution of trace
elements (50 mgL-1 MnCl24H2O, 5 mgL-1 H3BO3,
5 mgL-1 ZnCl2, 3 mgL-1 CuCl2, 1 mgL-1 Na2MoO4-
2H2O, 100 mgL-1 CoCl26H2O, 10 mgL-1 NiCl2-
6H2O, and 5 mgL-1 Na2SeO3). In the reactors,
330 mL of surfactant solution and 20 mL of sludge
were added to obtain a solid concentration of 1.5 gL-1
and a final volume of 350 mL. Surfactants solutions
were adjusted to pH 7.0 ± 0.2 and incubated in
darkness at 35°C ± 1 °C during 60 days. Net biogas
pressure (Dp) was automatically registered and trans-
duced to mass net carbon produced as gas in the
headspace (mh) using the gas lwas equation (Eq. 1).
1200  0:1ðDp  Vh Þ
mh ¼ ð1Þ
RT
where Dp is the difference between initial and final
pressures in the test reactor minus the corresponding in
the blank, and Vh is the volume of headspace in the
Table 2 Characteristic parameters of the inoculum
Parameter Value
pH 7.27
Total solids, g L-1 25.05
Volatile acidity, ppm CH3COOH 252
Alkalinity, ppm CaCO3 2500
Rate volatile acidity/alkalinity 0.10
Biodegradation (2017) 28:303–312
vessel (0.16 L).The validity of the test was checked
using sodium benzoate as reference (ultimate
biodegradation [60%).
Kinetic models
Biogas production was fitted to two kinetic models: (i)
pseudo first-order model, and (ii) logistic model.Using
the pseudo first-order model proposed by Larson
(1980, 1984), the biodegradation can be followed by
the concentration of the product generated (mh)
(Eq. 2).
 
mh ¼ a  1  eKðtcÞ ð2Þ
where mh is the amount of the product formed, K is a
rate constant which involves all the metabolic process
in the system, a is the maximum product generated,
and c is referred to the lag phase before the biodegra-
dation starts.The logistic model is based on the
Verhulst logistic function (Verhulst 1845, 1847)
(Eq. 3), where a is the maximum product formed, K
is the global rate constant and b is a constant of the
model.
a
mh ¼ ð3Þ
1 þ b  eKt
Results and discussion
Anaerobic biodegradation and inhibitory effects
High surfactant concentrations are usually found in
personal care product industry liquid wastes (Aham-
mad et al. 2013) and wastewater from laundry
([1000 mg L-1) (Braga and Varesche 2011). There-
fore, we studied the anaerobic biodegradability of
amine-oxide-based surfactants at the highest carbon
concentration (100 mg L-1 organic carbon) recom-
mended in the standard method, which implies initial
surfactant concentrations between 440.6 and
454.3 mg L-1, and approximately 0.30 g surfactant
g-1 dry sludge. Figure 2 shows the course of the net
biogas pressure Dp in the duration of the test.
As can be seen in Fig. 2, AO-R12 and AO-R14
showed negative values of the net biogas pressure
from 0.7 and 3 days (Fig. 2), i.e. production of biogas
was lower than in the blank digesters. This fact implies
307
that both surfactants, AO-R12 and AO-R14, inhibit
endogenous biogas production practically from the
beginning of the biodegradation test. The inhibition
can be calculated by measuring the final cumulated
pressure in the test samples Dpf related to final
cumulated pressure in the blanks Dpb (Eq. 4). Inhibi-
tion percentages and confidence intervals (95%) are
given in Table 3, and are around 90% for both
surfactants.
 
Dpf
Ih ð% Þ ¼ 1   100 ð4Þ
Dpb
Inhibitory effects of surfactants on anaerobic
biodegradation processes have been extensively
reported. For example, Delforno et al. (2015) found
that linear alkylbenzenesulfonate (LAS) at concentra-
tion greater than 20 mg L-1 causes the inhibition of
the anaerobic microbiota and Rı́os et al. (2016)
reported the inhibition of methanogenic microorgan-
isms in the anaerobic biodegradation of alkylpolyglu-
cosides. In case of amine oxides, Garcı́a et al. (2007)
found a significate inhibition of endogenous biogas
production, although they carried out tests at lower
initial concentrations (15–275 mg L-1 organic car-
bon) and lower ratio mg surfactant g-1 dry sludge
(5–80).
Studies on anaerobic biodegradation show consid-
erable variation in the inhibition mechanisms reported
for most compounds. The main reason for these
variations is the complexity of the anaerobic biodegra-
dation process where mechanisms such as antagonism,
synergism, acclimation, and complexing could signif-
icantly affect the phenomenon of inhibition. The
accumulation of surfactants in bacterial membranes
causes the membrane to swell and leak, disrupting ion
gradients and eventually causing cell lysis (Chen et al.
2008). According to a previous study, the actual mode
of action (MoA) of amine-oxide-based surfactants is
unknown, although most probably, it is not class 1
narcosis due to their amphoteric character (Rı́os et al.
2017a). Accumulation of these surfactants may cause
reactor upset, as indicated by reduced biogas produc-
tion and possible reactor failure.
On the other hand, net biogas pressure from
anaerobic biodegradation of AO-cocoamido increased
continuously until around 22 days and remained
constant for the rest of the assay, indicating the end
of the anaerobic process. Figure 2 shows that there is
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308 Biodegradation (2017) 28:303–312

Fig. 2 Time course of the 250

net biogas pressure
200

Net biogas pressure (mbar)

150
AO-Cocoamido
100
AO-R12

50 AO-R14

-50

-100

-150
0 5 10 15 20 25 30 35 40 45 50 55 60
t, days

Table 3 Characteristic parameters of the anaerobic biodegradation of amine-oxide-based surfactants (95% CI)
Surfactant Ih (%) Dh (%) Dt (%) VM (%/days)

AO-R12 91.7 ± 2.2 – – –

AO-R14 89.4 ± 3.0 – – –
AO-Cocoamido \0 39.4 ± 8.0 60.8 ± 2.2 1.8 ± 0.6

no lag-phase; from the beginning of test the net biogas
pressure rises, first rapidly, until the 12th day, and then
slower until the 22th day. The absence of the
adaptation phase contrasts to the results obtained by
Garcı́a et al. (2007) in the anaerobic biodegradation of
AO-cocoamido, in which the methanogenic popula-
tion needed about 3 and 6 weeks to initiate the biogas
production. Production of biogas in the AO-co-
coamide tests was higher than biogas production in
blank digesters. Ih-AO-cocoamido showed a negative
value (Table 3), i-e. AO-cocoamido did not inhibit
endogenous biogas production.
Differences in biogas production between amine-
oxide-based surfactants can be explained by their
structural differences, the presence of an amine group
in the molecule of AO-cocoamido increases its
hydrophilic character. These differences are also
reflected in its CMC, which is approximately 1.9 and
2.9 times higher than CMC of AO-R12 and AO-R14
respectively, and in their toxicity to several aquatic
organisms, for which AO-cocoamido is considerably
less toxic (Fernández-Serrano et al. 2014; Lechuga
et al. 2016). According to Wong et al. (1997) and
Jurado et al. (2009) surfactants with higher hydro-
philic-lipophilic balance (HLB) are less toxic, and
therefore could be less harmful to the methanogenic
microorganisms from the sludge.
The above mentioned differences could also be
explained focusing on the biodegradation pathway. It
is well recognized in the literature that, microbial
degradation of amine-oxides proceeds via two path-
ways: (a) x-oxidation on the terminal methyl group to
obtain x-amino fatty acid, which is followed by a
deamination yielding amine and x-oxo fatty acid as
intermediates or (b) initial fission of the Calkyl-N bond
through a degradation reaction, forming alkanals and
fatty acids as intermediates. In the absence of oxygen,
almost only the Calkyl-N fission can take place (Garcı́a
et al. 2007; Rı́os et al. 2017b). Therefore, Calkyl-N
fission is considered the main biodegradation path-
way. This fact agrees with the higher biodegradation
of the amine-oxide-based surfactant having an amide
group in the alkyl chain (AO-Cocoamido), which is
not present in AO-R12 and AO-R14. In biodegradation

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Biodegradation (2017) 28:303–312 309

of AO-Cocoamido, after the cleavage of the C–N from
the amide group, a carboxyl is generated, which speed
up the biodegradation process if compared with AO-
R12 and AO-R14. On the other hand, the cleavage of
the C–N bond of the amine requires stronger condi-
tions which make it a more difficult site to be attacked.
This reinforces the obtained results.
Time course of the mass net carbon produced as gas
(mh) during the anaerobic process of AO-cococamido
is shown in Fig. 3. Most of the carbon produced as gas
was obtained during the first fifteen days. The
maximum amount was 13.78 mg organic carbon,
representing the 39.4% of biodegradation (Dh) calcu-
lated from the headspace gas using Eq. (5), where mv
is the mass of the test substance carbon (Table 3).
 
mh
D h ð% Þ ¼  100 ð5Þ
mv
Taking into account the inorganic carbon dissolved in
the liquid phase at the end of the assay (mi=7.48 mg),
the amount of total gasified carbon (mt) was 21.26 mg,
which means a final mineralization (Dt) of 60.8%
(Table 3) calculated according to Eq. (6).
 
mt
D t ð% Þ ¼  100 ð6Þ
mv
According to the Organization for Economic Co-
operation and Development Guidelines for the Testing
of Chemicals (OECD 311 2006), a complete anaerobic
biodegradation can be assumed if 75–80% of the
theoretical gas production is overtaken, i.e. if Dt is
higher than 75–80%. However, AO-cocoamido can be
considered biodegradable at the initial concentration
of 100 mg L-1 organic carbon if the Eco-Label criteria
are considered (biodegradation rate [60%) (EC
66/2010).
The mean velocity of biodegradation (VM,
%/day) can be calculated from the degree of
biodegradation from the headspace (Dh), as the
mean velocity of biodegradation reached until
achieving 15% biodegradation of the surfactant
(Jurado et al. 2013). This parameter informs about
the quickness of the biodegradation and permit to
compare the anaerobic biodegradability of surfac-
tants. Table 3 shows VM values calculated and its
confidence intervals (95%). This parameter reveals
the slower biodegradation of the AO-cocoamido
when compared to other surfactants such as alkyl
polyglucusides for which the parameter VM ranged
from 2.01 to 4.86%/day (Rı́os et al. 2016). How-
ever, the degree of biodegradation calculated from
the headspace (Dh) and the final mineralization (Dt)
of AO-cocoamido reached a higher level than the
three alkyl polyglucosides studied by Rı́os et al.
(2016), for which Dh ranged from 19.3 to 32.8%
and Dt from 19.6 to 40%.
Amine-oxide-based surfactants are rapidly
biodegradable under aerobic conditions (OECD SIDS
2006, Garcı́a et al. 2007; Jurado et al. 2013) but in
anaerobic conditions their biodegradation might be
impeded. The anaerobic digestion of a substance until
its complete mineralization is a convoluted process of

Fig. 3 Time course of the 16.00

mass net carbon produced as
gas in the anaerobic 14.00
biodegradation of AO-
cocoamido 12.00

10.00
mh, mg

mh
8.00
Pseudo first-order
6.00 Logistic

4.00

2.00

0.00
0 5 10 15 20 25 30 35 40 45 50 55 60
t, days

123
310
Table 4 Kinetic parameters of anaerobic biodegradation of AO-cocoamido
Kinetic model a (mg) c (days)
Pseudo first-order 14.10 1.45
Logistic 13.61 – 19.32
understanding reactions by the co-operation of several
micro-organisms. They work sequentially, one organ-
ism consumes as substrate the metabolites produced
by the previous one. For this reason the anaerobic
mineralization takes place slower and the surfactant
reaches lower final percentages of degradation than
during the aerobic biodegradation (Rı́os et al. 2017b).
Anaerobic biodegradation kinetics
Kinetics of the anaerobic biodegradation was only
studied for AO-cocoamido since AO-R12 and AO-R14
showed negative values of the net biogas pressure and
mass net carbon produced as gas (mh) could not be
calculated. Due to the complexity of the biological
process taking place in an anaerobic waste water
treatment reactor is difficult to identify the individual
reactions. Pseudo first-order model was used in a
previous work in order to model the anaerobic kinetics
of biodegradation of alkyl polyglucosides (Rı́os et al.
2016). The time course of the mass net carbon
produced as gas in the anaerobic biodegradation of
AO-cocoamido was also fitted to a pseudo first-order
equation Eq. (2) (Fig. 3). The kinetic parameters are
shown in Table 4.
In this case, there are some deviations from the
model proposed in the first thirty days, because the
product generation follows a logistic increase; the lag
time and the maximum product formed were not well
established using this model. For this reason, we also
used the kinetic analysis method of growth curves
based on the Verhulst logistic function (Eq. 3).
Figure 3 and Table 4 show the representation and
the kinetic parameters respectively.
The logistic model fits remarkably well the biogas
production. The parameter a reflects accurately the
maximum product formed and residuals are not above
0.7 in any case, therefore this modelling could be used
to evaluate the biodegradation of AO-cocoamido
under anaerobic conditions.
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Biodegradation (2017) 28:303–312
b K (days-1) r
– 0.1024 0.9820
0.3158 0.9978
Conclusions
Results of this study confirm that some amine-oxide-
based surfactants are not completely biodegradable in
anoxic conditions at the selected standard concentra-
tion of 100 mg L-1 organic carbon. AO-cocoamido
showed a rapid biodegradation in the first half of the
test and did not cause inhibition of biogas production.
Otherwise, AO-R12 and AO-R14 inhibited the biogas
production from the beginning of the test, inhibition
percentages were around 90%; therefore anaerobic
biodegradation of these surfactants was probably
hampered by the toxic effects on the heterogeneous
bacteria mixture. Differences in the biodegradability
can be explained by the higher hydrophilic character
of AO-cocoamido as consequence of the amide group
in its molecule. In addition, the continuous measure-
ment of the carbon generated as gas made possible to
fit considerably well the biodegradation kinetics to a
logistic model and kinetic parameters were also
determined.
Acknowledgements The Spanish Ministry of Economy and
Competitiveness is gratefully acknowledged for having funded
the project CTQ2015-69658-R.
References
Ahammad SZ, Bereslawski JL, Dolfing J, Mota C, Graham DW
(2013) Anaerobic-aerobic sequencing bioreactors improve
energy efficiency for treatment of personal care product
industry wastes. Bioresour Technol 139:73–79. doi:10.
1016/j.biortech.2013.03.185
APHA-AWWA-WPCF (1992) Standard methods for the
examination of water and wastewater, 18th edn. American
Public Health Association, Washington, DC
Baena-Nogueras RM, González-Mazo E, Lara-Martı́n PA
(2013) Determination and occurrence of secondary alkane
sulfonates (SAS) in aquatic environments. Environ Pollut
176:151–157. doi:10.1016/j.envpol.2013.01.020
Baena-Nogueras RM, Rojas-Ojeda P, Sanz JL, González-Mazo
E, Lara-Martı́n PA (2014) Reactivity and fate of secondary
alkane sulfonates (SAS) in marine sediments. Environ
Pollut 189:35–42. doi:10.1016/j.envpol.2014.02.019
Biodegradation (2017) 28:303–312
Baglioni M, Jàidar Benavides Y, Berti D, Giorgi R, Keiderling
U, Baglioni P (2015) An amine-oxide surfactant-based
microemulsion for the cleaning of works of art. J Colloid
Interface Sci 440:204–210. doi:10.1016/j.jcis.2014.10.003
Braga JK, Varesche MBA (2011) Commercial laundry water
characterization for anaerobic treatment in fluidized bed
reactor. X DAAL—X Latin American workshop and
symposium on anaerobic digestion, Ouro Preto
Chen Y, Cheng JJ, Creamer KS (2008) Inhibition of anaerobic
digestion process: a review. Bioresour Technol
99:4044–4064. doi:10.1016/j.biortech.2007.01.057
Colmenar-Santos A, Zarzuelo-Puch G, Borge-Diez D, Garcı́a-
Diéguez C (2016) Thermodynamic and exergoeconomic
analysis of energy recovery system of biogas from a
wastewater treatment plant and use in a Stirling engine.
Renew Energy 88:171–184. doi:10.1016/j.renene.2015.11.
001
Delforno TP, Moura AGL, Okada DY, Sakamoto IK, Varesche
MBA (2015) Microbial diversity and the implications of
sulfide levels in an anaerobic reactor used to remove an
anionic surfactant from laundry wastewater. Bioresour
Technol 192:37–45. doi:10.1016/j.biortech.2015.05.050
Delforno TP, Okada DY, Faria CV, Varesche MBA (2016)
Evaluation of anionic surfactant removal in anaerobic
reactor with Fe(III) supplementation. J Environ Manag.
doi:10.1016/j.jenvman.2016.09.026
Dereszewska A, Cytawa S, Tomczak-Wandzel R, Medrzycka K
(2015) The effect of anionic surfactant concentration on
activated sludge condition and phosphate release in bio-
logical treatment plant. Polish J Environ Stud 24:83–91.
doi:10.15244/pjoes/28640
EC 66/2010. European Parliament Regulation (EC) No.
1980/2000 of the European Parliament and of the Council
of 25 November 2009 on the EU Ecolabel. Official Journal
of the European Union. L 27/1, 30 Jan 2010
Fernández-Serrano M, Jurado E, Fernández-Arteaga A, Rı́os F,
Lechuga M (2014) Ecotoxicological assessment of mix-
tures of ether carboxylic derivative and amine-oxide-based
non-ionic surfactants on the aquatic environment. J Sur-
factants Deterg 17:1161–1168. doi:10.1007/s11743-014-
1621-2
Garcı́a MT, Campos E, Ribosa I (2007) Biodegradability and
ecotoxicity of amine oxide based surfactants. Chemo-
sphere 69:1574–1578. doi:10.1016/j.chemosphere.2007.
05.089
Huang P, Goel R (2015) Response of a sludge-minimizing lab-
scale BNR reactor when the operation is changed to real
primary effluent from synthetic wastewater. Water Res
81:301–310. doi:10.1016/j.watres.2015.04.035
ISO 11734:1995. Standard ISO 11734:1995. Water quality.
Evaluation of ultimate anaerobic biodegradability of
organic compounds in digested sludge. Method by mea-
surement of biogas production
Jenkins D, Wanner J (2014) Activated sludge—100 years and
counting. IWA publishing, London
Jurado E, Fernández-Serrano M, Núñez-Olea J, Luzón G,
Lechuga M (2009) Acute toxicity and relationship between
metabolites and ecotoxicity during the biodegradation pro-
cess of non-ionic surfactants: fatty-alcohol ethoxylates,
nonylphenol polyethoxylate and alkylpolyglucosides. Water
Sci Technol 59:2351–2358. doi:10.2166/wst.2009.266
311
Jurado E, Fernández-Serrano M, Rı́os F, Lechuga M (2013).
Aerobic biodegradation of surfactants. In: Chamy R,
Rosenkranz F (eds) Biodegradation—life of science, In-
Tech, Rijeka, pp 66–81. doi: 10.5772/56120
Kapanen A, Vikman M, Rajasärkkä J, Virta M, Itävaara M
(2013) Biotests for environmental quality assessment of
composted sewage sludge. Waste Manag 33:1451–1460.
doi:10.1016/j.wasman.2013.02.022
Larson R (1980) Role of biodegradation kinetics in predicting
environmental fate. In: Dickson KJ, Cairns J, Maki AW
(eds) Biotransformation and fate of chemicals in the
aquatic environment. American Society of Microbiology,
Washington DC, pp 67–86
Larson RJ (1984) Kinetics and ecological approaches for pre-
dicting biodegradation rates of xenobiotic organic chemi-
cals in natural eco-system. In: Klug MJ, Reddy CA (eds)
Current perspectives in microbial ecology. American
Society of Microbiology, Washington DC, pp 677–686
Lay CH, Huang CY, Chen CC, Lin CY (2016) Biohydrogen
production in an anaerobic baffled stacking reactor: recir-
culation strategy and substrate concentration effects. Bio-
chem Eng J 109:59–64. doi:10.1016/j.bej.2016.01.001
Lechuga M, Fernández-Arteaga A, Fernández-Serrano M, Jur-
ado E, Burgos A, Rı́os F (2014) Combined use of ozonation
and biodegradation of anionic and non-ionic surfactants.
J Surfactants Deterg 17:363–370. doi:10.1007/s11743-
013-1480-2
Lechuga M, Fernández-Serrano M, Jurado E, Núñez-Olea J,
Rı́os F (2016) Acute toxicity of anionic and non-ionic
surfactants to aquatic organisms. Ecotoxicol Environ Saf
125:1–8. doi:10.1016/j.ecoenv.2015.11.027
Lettinga G (2001) Challenge of psychrophilic anaerobic
wastewater treatment. Trends Biotechnol 19:363–370.
doi:10.1016/S0167-7799(01)01701-2
Liwarska-Bizukojc E, Bizukojc M (2008) Estimation of the
impact of anionic surfactants on activated sludge flocs
morphology in a batch system. Arch Environ Prot
34:25–34
Martı́nez-Garcı́a CG, Fall C, Olguı́n MT (2016) Activated
sludge mass reduction and biodegradability of the
endogenous residues by digestion under different aerobic
to anaerobic conditions: comparison and modeling.
Bioresour Technol 203:32–41. doi:10.1016/j.biortech.
2015.12.015
McCarty PL, Bae J, Kim J (2011) Domestic wastewater treat-
ment as a net energy producer–can this be achieved?
Environ Sci Technol 45:7100–7106. doi:10.1021/
es2014264
OECD 311 (2006) OECD Guidelines for the Testing of
Chemicals. 3. Degradation and accumulation. Test number
311: Anaerobic biodegradability of organic compounds in
digested sludge: by measurement of gas production
OECD SIDS (2006) Screening information dataset initial
assessment report for SIAM 22. Amine Oxides. Paris,
France, 18–21 April 2006
Pedrazzani R, Cerett E, Zerbini I, Casale R, Gozio E, Bertanza
G, Gelatti U, Donato F, Feretti D (2012) Biodegradability,
toxicity and mutagenicity of detergents: Integrated exper-
imental evaluations. Ecotoxicol Environ Saf 84:274–281
Richter E, Roller E, Kunkel U, Ternes TA, Coors A (2016)
Phytotoxicity of wastewater-born micropollutants—
123
312
characterisation of three antimycotics and a cationic sur-
factant. Environ Pollut 208:512–522. doi:10.1016/j.
envpol.2015.10.024
Rı́os F, Fernández-Arteaga A, Lechuga M, Jurado E, Fernández-
Serrano M (2016) Kinetic study of the anaerobic
biodegradation of alkyl polyglucosides and the influence of
their structural parameters. Environ Sci and Pollut Res
23:8286–8293. doi:10.1007/s11356-016-6129-z
Rı́os F, Fernández-Arteaga A, Lechuga M, Fernández-Serrano
M (2017a) Ecotoxicological characterization of poly-
oxyethylene glycerol ester non-ionic surfactants and their
mixtures with anionic and non-ionic surfactants. Environ
Sci Pollut Res 24:10121–10130. doi:10.1007/s11356-017-
8662-9
Rı́os F, Lechuga M, Fernández-Serrano M, Fernández-Arteaga
A (2017b) Aerobic biodegradation of amphoteric amine-
oxide-based surfactants: Effect of molecular structure,
initial surfactant concentration and pH. Chemosphere
171:324–331. doi:10.1016/j.chemosphere.2016.12.070
Sharvelle SE, Garland J, Banks MK (2008) Biodegradation of
polyalcohol ethoxylate by a wastewater microbial consor-
tium. Biodegradation 19:215–221. doi:10.1007/s10532-
007-9128-4
Singh SK, Bajpai M, Tyagi VK (2006) Amine oxides: a review.
J Oleo Sci 55:99–119. doi:10.5650/jos.55.99
Soares A, Guieysse B, Jefferson B, Cartmell E, Lester JN (2008)
Nonylphenol in the environment: a critical review on
occurrence, fate, toxicity and treatment in wastewaters.
Environ Int 34:1033–1049. doi:10.1016/j.envint.2008.01.
004
Verhulst P-F (1845) Recherches mathématiques sur la loi
d’accroissement de la population. Nouv. mém. de
123
Biodegradation (2017) 28:303–312
l’Academie Royale des Sci. et Belles-Lettres de Bruxelles
18:1–41
Verhulst P-F (1847). Deuxième mémoire sur la loi d’ac-
croissement de la population. Mém. de l’Academie Royale
des Sci., des Lettres et des Beaux-Arts de Belgique 20:
1–32
Wang Y, Wang Q, Li M, Yang Y, He W, Yan G, Guo S (2016)
An alternative anaerobic treatment process for treatment of
heavy oil refinery wastewater containing polar organics.
Biochem Eng J 105:44–51. doi:10.1016/j.bej.2015.08.012
WD (2000) Working Document on Sludge (3rd Draft), Com-
mission of the European Communities; Directorate-Gen-
eral Environment, ENV.E.3/LM, Brussels, 27th April 2000
WD (2010) Working Document: Sludge and Biowaste, Euro-
pean Commission; Directorate-General Environment; DG
ENV C2/BZ/tb, Brussels, 21st September 2010
Wong DCL, Dorn PB, Chai EY (1997) Acute toxicity and
structure–activity relationships of nine alcohol ethoxylate
surfactants to fathead minnow and daphnia magna. Environ
Toxicol Chem 16:1970–1976. doi:10.1897/1551-5028
(1997)016\1970:ATASAR[2.3.CO;2
Zhang C, Cui F, Zeng G, Jiang M, Yang Z, Yu Z, Zhu M, Shen L
(2015) Quaternary ammonium compounds (QACs): a
review on occurrence, fate and toxicity in the environment.
Sci Total Environ 518–519:352–362. doi:10.1016/j.
scitotenv.2015.03.007
Zhou A, Liu W, Varrone C, Wang Y, Wang A, Yue X (2015)
Evaluation of surfactants on waste activated sludge fer-
mentation by pyrosequencing analysis. Bioresour Technol
192:835–840. doi:10.1016/j.biortech.2015.06.017