Thesis Nan Wang
Thesis Nan Wang
Thesis Nan Wang
Activated Sludge
Thesis by
Nan Wang
Master of Science
June 2013
2
ABSTRACT
Nan Wang
The study examined the removal mechanism of non-acclimated and acclimated aerobic
activated sludge for 29 target organic micropollutants (OMPs) at low concentration. The
vaporization. Adsorption and biodegradation were found to be the main routes for OMPs
removal for all target OMPs. Target OMPs responded to the two dominant removal
routes in different ways: (1) complete adsorption, (2) strong biodegradation and weak
adsorption, (3) medium biodegradation and adsorption, and (4) weak sorption and weak
propylparaben well followed first-order model (R2: 0.939 to 0.999) with the rate
constants ranging from 0.519-7.092 h-1. For biodegradation kinetics, it was found that
followed zero-order model (K0:1.15E-4 to 0.0142 µg/Lh-1, R2: 0.991 to 0.999), while
Inhibition by sodium azide (NaN3)and high temperature sterilization was compared, and
it was found that high temperature sterilization will damage cells and change the sludge
charge state.
For the OMPs adaptation removal study, it was found that some of OMPs effluent
concentration decreased, which may be due to the slow adaptation of the sludge or the
increase of certain bacteria culture; some increased due to chromic toxicity of the
chemicals; most of the OMPs had stable effluent concentration trend, it was explained
that some of the OMPs were too difficutl to remove while other showed strong quick
adaptation.
membrane filtration (NF-MBR) was developed to further study the OMPs removal and to
exam the concept of compounds (CRT). The NF-MBR was proved to be a promising
concentration in permeate water, the apparent removal rate was over 80% for most of the
OMPs.
5
ACKNOWLEDGEMENTS
I would like to thank my committee chair, Dr. Gary Amy, and my committee members,
Dr. Peiying Hong, Dr. Tao Zhang, for their guidance and support throughout the course
of this research.
I would like also thank Dr. Chunhai Wei for his patient help on the experimental design
and operation of the reactors. Thank Dr. Christiane Hoppe Jones for her helps on the
My appreciation also goes to my friends and colleagues and the department faculty and
staff for making my time at King Abdullah University of Science and Technology a great
experience.
TABLE OF CONTENTS
ABSTRACT…………………………………………………………………………………………………………….....…..3
ACKNOWLEDGEMENTS………………………………………………..…………………………………………….…5
LIST OF TABLES……………………………………………………………………………………….……….……………13
2.1 Introduction............................................................................................................. 20
3.1 Introduction............................................................................................................. 27
inhibition .................................................................................................................... 43
4.1 Introduction............................................................................................................. 50
5.1 Introduction............................................................................................................. 57
5.1.2 Methods............................................................................................................ 58
6.1 Conclusions.............................................................................................................. 67
6.2 Suggestions.............................................................................................................. 68
References…………………………………………………………………………………………………………..……….69
Appendix A ………………………………………….…………………………………………………………………..…74.
Appendix B ………………………………………….………………………………………………………………….….76.
Appendix A ………………………………………….………………………………………………………………..……78.
Appendix D ………………………………………….……………………………………………………..……………..86.
Appendix E ………………………………………….………………………………………………………………….…..92
10
LIST OF ILLUSTRATIONS
Figure 3-4. The removal of sulfamethoxazole by aerobic activated sludge. A total removal,
Figure 3-5. The removal of propylparaben by aerobic activated sludge. A total removal, B
Figure 4-3. Removal trend of diclofenac (A) and iopromide (B) ........................................ 53
12
Figure 4-4. Removal trend of atenolol (A) and trimethoprim (B) ....................................... 54
Figure 4-5. Removal trend of clofibric acid (A) and benzefibrate (B) ................................ 55
Figure 4-6. Removal trend of DEET, clofibric acid, oxybenzone, sulfamethoxazole, TCEP
and dilatin............................................................................................................................. 56
Figure 5-3. Cin, Cp and CRT changes of acesulfame in NF-SB-MBR operation ................ 63
Figure 5-4. Relation between CRT and rejection rate in acefulfame .................................. 64
Figure 5-5. Cin, Cp and CRT changes of bisphenol A in NF-SB-MBR operation ............... 64
Figure 5-6. Cin, Cp and CRT changes of amtripline in NF-SB-MBR operation .................. 65
LIST OF TABLES
Table 2-3. Operation parameters of the aerobic activated sludge bioreactor ...................... 26
Table 3-1. Summary of target organic micropollutants and their physic-chemical properties.
.............................................................................................................................................. 29
Table 3-4. COD changes between 0h and 168h in the adsorption and biodegradation
experiment............................................................................................................................ 38
Table 3-5. First-order adsorption kinetics rate constants of target OMPs ........................... 44
Table 3-8. OMPs removal rate comparison between literature and the study ..................... 48
Chapter 1. Introduction
attracted great concerns [1] [2]. Traditional water treatment processes were not designed
for the micropollutants removal, although some reports have demonstrated that some
such as activated sludge. These chemicals are usually at low concentration (ng/L-µg/L) [3,
4], but they cannot be negligible as they may still course serious effect on human health
and the environment. Many PPCPs possess acute toxicities similar to industrial
chemicals, exhibit chronic toxicity ,and exhibit ecotoxic effects principally through
endocrine disruption [5]. Concern is also mounting that low level contamination by
certain PPCPs may contribute to the spread of antibacterial resistance [6]. To date, more
than 80 different compounds have been detected in surface and ground water worldwide.
The disposal of unused medication via the toilet seems to be of minor importance but
many of the pharmaceuticals applied in human medical care are not completely
eliminated in the human body [2]. These micropollutants are usually released into the
aquatic environment by effluents from municipal sewage treatment plants (STPs), and
investigations have shown that these substances are often not eliminated during waste
water treatment and also not biodegraded in the environment [2, 7, 8]. The low
sorption to biomass or suspended solid [9, 10]. Among these, sorption processes and
oxidation processes will be the most effective removal strategy. Although advanced
oxidation processes require more capital and operation costs, they are able to oxidize
many organic micropollutants, and either eliminate them completely or make them
People in private households are either dumping drugs in normal metabolism routes or
dispose expired drugs into the toilets [7, 12]. Sometimes, hospital wastewater can
contribute to the load due to unregular operations. Some organic micropollutants such
EDCs and PPCPs are not biodegradable in the WWTPs, and the domestic sewage will be
then disposed into the natural environment. Another route for the discharge of
micropollutants is by the usage of green fertilizer. When manure is used, it is very likely
that the veterinary pharmaceuticals been introduced into the rivers and groundwater by
runoff. The other source from agriculture is from the usage of pesticides, which has long
been known. Point –source discharges by pharmaceutical plants are also released into the
rivers, which is always a serious problem for the local ecology. Leakage from landfills
can also contaminate waters, as some of landfills are not secure landfill [13]. Figure 1-1
shows the typical origin routes of pharmaceutical products, and Figure 1-2 shows
Some investigations have been carried out in Austria, Brazil, Canada, Croatia, England,
Germany, Greece, Italy, Spain, Switzerland, the Netherlands, and the U.S., and more than
80 compounds, pharmaceuticals and several drug metabolites have been detected in the
Figure 1-2. Transformation pathways of PPs (PC: parent compound, M: Metabolite(s), TP:
Transformation Product(s), WWTP: Waste water Treatment Plant; DWT: Drinking
WaterTreatments)[14]
In the recent reports, many researchers have found new environmental effects of the
PPCPs. However, because of the lack of assessment of complete data, it is difficult for
researchers to fully understand the environmental effects of most PPCPs, and no one
knows whether the relatively low concentrations found for PPCPs produce adverse
effects on biota or whether the toxicity of complex mixtures might be totally different
18
from that of individual compound [7, 16, 17]. Thus, we need more research and
generation and disposal into the environment. Fortunately, there are many research
institutes and government agencies that are intensively working on the topic. The EU and
the U.S. have launched research projects like EU’s Repharmawater and Poseidon and U.S.
Analgesics and anti-inflammatory drugs are often prescribed in human medical care but
often they are sold at higher quantities as over the counter (OTC) drugs. In Germany, the
total quantities of acetaminophen (ASA) sold per year have been estimated at over 500
tons [20, 21]. Other analgesics such as diclofenac or ibuprofen are sold in Germany at an
annual amount of about 75 and 180 tons respectively [21]. The concentration of ASA is
rather low in the sewage effluents and surface water. Ternes et al. [8] found that the
sewage influents are 54, 6.8 and 4.6 µg/L, respectively, and all of these three compounds
were efficiently removed by municipal STPs. Another pain killer, acetaminophen is also
easily degraded and removed by STPs, and seldom has acetaminophen been detected [22].
Several researches have been done to investigate the occurrence of antibacterial drugs in
STPs and surface waters [23, 24]. Macrolide antibiotics (clarithromycin, dehydro-
and trimethoprim have been found up to the low µg/l-level in sewage and surface water
samples [15]. Penicillin and tetracyclines were not detected which was explained that
19
penicillin are easily hydrolyzed and tetracyclines readily precipitated with cations such as
calcium and accumulate in sewage sludge or sediments [25, 26]. However, antibiotics are
detected at high concentration in hospital wastewater and primary and tertiary wastewater
effluents [27].
The data collected above indicate that some PhACs are originated from municipal
WWTPs as they cannot be completely eliminated. Most of them are at low level
concentration up to µg/L in municipal sewage and surface water. New methods to detect
Some pharmaceuticals have been found to be EDCs, and it has long been known that
EDCs can cause unpredictable side effects especially in central nervous system [28] and
hormone systems. Thus, the concern of the occurrence of EDCs is rapidly increasing.
Generally, EDCs are classified as estrogenic, androgenic and thyroidal [29]. EDCs have
been detected by several cities at wide ranges of concentration. The removal efficiency of
different operation units is not clear, but most removal seems to be done within the
activated sludge process. The removal efficiency of activated sludge is better than that of
2.1 Introduction
The conventional activated sludge (CAS) process is the mostly widely used wastewater
treatment system in the world; therefore, the study of the removal organic micropollutants
(OMPs) by CAS is of vital importance. In this study, conventional aerobic activated sludge
wastewater treatment was chosen for the sludge operation, and followed a classical
sequencing batch reactor mode. In order to get the non-acclimated activated sludge,
synthetic wastewater without any OMPs content was used as influent. After 2 weeks of
operation, the effluent water reached equilibrium, and the reactor was kept operating for
about 6 months. All of the parameters of the effluent reached steady state after two weeks.
The sludge could be used for the studying of OMPs removal by non-acclimated aerobic
activated sludge.
The Applikon ez-control system, which is shown in Figure 2-1, was used as the reactor for
the culture of non-acclimated activated sludge. Parameters like temperature, pH, DO, ORP
could be controlled or monitored by the system, and influent and effluent were also control
by the built-in pump. The sludge container was made of double glazing glass with a total
volume of 2.4 L, the temperature could be controlled by circulating water in the glass
interlayer. All of the pipes were covered with aluminum foil to inhibit the growth of algae.
21
The composition of the synthetic wastewater was chosen according to Nopens et al.[30].
All the chemicals are purchased from Sigma-Aldrich, which are listed in Table 2-1. The
theoretical COD was 439.47 mg/L, the measured COD was around 380-410mg/L. The
possible reason for the low value was the loss of large molecules after filtration of the
The seeding sludge was taken from the Jeddah wastewater plant, and was filtered through
100 µm sieve.
22
The operation of the reactor followed a classical sequencing batch reactor mode, which is
Fill mode: 1L synthetic waste water was filled into the reactor through the pipe, and the
flow rate was 2L/h, and the filling time was 30 mins, after filling, the water reached the
React mode: Immediately after filling, the valve of pure oxygen was open, and oxygen
was bubbled into the reactor under a pressure of 0.01 bar for two hours. DO was 75-81%
under constant temperature of 20℃. Meanwhile, the built-in stirring started at the speed of
200 rpm.
Settle mode: The settling time was 2 hours, while the stirrer and aeration was stopped.
Draw mode: Effluent rate was also 2L/h and draw for 30 mins, the effluent was collected
During the 6 month operation of the reactor, COD, TOC, pH, DO, VFA, MLSS, MLVSS,
TN, NO3-N, NH3-N, and other parameters were intensively monitored. All samples were
COD, VFA, TN, NO3-N, NH3-N: Hach series parameter reagents were used to measure
the above parameters. A Hach DBR 200 digital block reactor was used for digestion, and a
MLSS: MLSS was measured using the standard method. The filters used were Whatman
0.45 µm filters, the volume used for sludge was 10 mL and for effluent was 200 mL, the
m1 m0
MLSS (1.1)
V
Where,
MLVSS: The mixed liquor volatile suspended solids (MLVSS) represented the population
size of bacteria within the activated sludge process. Volatile suspended solids were solids
that burn in a muffle furnace at 550°C for 3 hours, where the loss of volatile solids was
m1 m2
MLSS (1.2)
V
SVI: SVI is the sludge volume index, which was used to measure the settling character.
SVI was the volume of the MLSS over the density of the mixed liquor suspended solids.
Influent
2.2L 2.2L
Effluent
1.2L
The starting parameters for the bioreactor are shown in Table 2-2. The operation was kept
the same for about 6 months, and the bioreactor effluent was stable during the process.
The effluent samples were taken and analyzed every few days. Paraeters are shown
in Table 2-3. COD and TOC could be removed over 95%, and 80% for organic nitrogen at
most.
26
2.4 Summary
The aerobic activated sludge was operated for 6 months, and the effluent was stable. No
OMPs were spiked into the synthetic wastewater, therefore, the sludge could be considered
as “non-acclimated sludge” for OMPs, and could be used for further studies.
27
3.1 Introduction
In this chapter, the removal mechanisms of 29 selected organic micropollutants (OMPs)
with different chemical and physical properties were investigated using batch reactors. The
work studied the roles of adsorption, biodegradation, hydrolysis and volatilization in the
removal of target OMPs, it was found that hydrolysis and volatilization effects could be
considered negligible, while adsorption and biodegradation played the main roles in the
process. To study the adsorption route, NaN3 was used as an inhibitor, and it was
demonstrated that it worked well as expected, as COD as observed not changed after the
NaN3 injection. The removals could be classified according to different routes: strong
3.2.1 Materials
We selected 29 OMPs as target for their removal in the study, and they were typical OMPs
reported in the literature. The selection considered the differences between physic-
chemical properties, like hydrophobicity, charge state, as well as their classification and
usage.
Acesulfame, atrazine, caffeine, DEET, TCEP, TCPP, TDCPP, were purchased from Fluka
propylparaben were purchased from CDN, isotopes of atrazine, ibuprofen were purchased
from Fluka, and isotopes of benzafibrate, clofibric acid, oxybenzone, primidone, TCEP
were purchased from Cambridge Isotope Laboratories. All stock solutions were prepared in
methanol with a concentration of 1000 mg L-1, and stored at -20 ℃. Table 3-1 shows
asummary of target OMPs and their physic-chemical properties, and the structures of the
The synthetic waste water followed the recipe which was described in Chapter 2. Sodium
azide, which was used to inhibit the sludge biodegradation, was purchased from Sigma-
Aldrich (USA). All of the selected OMPs have been found in municipal water treatment
plants [2, 4, 5, 31-38]. As the acclimation of sludge can influence the degradation of OMPs
at trace concentration, the sludge used had been running without OMPs for 6 months,
which can ensure the sludge was in non-acclimated stages for OMPs.
29
Table 3-1. Summary of target organic micropollutants and their physic-chemical properties.
Compound CAS use Formula MW K Ha logKow pKa Kdb Charge
g/mol atm•m3/mol L/kgS
e S
Acesulfame 33665-90-6 Sweetener C4H5NO4S 163.15 9.63E-009 -1.33 -c
Amitriptyline 50-48-6 Antibiotic C20H23N 277.41 6.85E-008 4.92 9.4 2686 +
Atenolol 29122-68-7 Beta-blocker C14H22N2O3 266.34 1.37E-018 0.16 9.6 35 +
Atrazine 1912-24-9 Pesticide C8H14ClN5 215.68 2.36E-009 2.61 1.7 60 N
Bezafibrate 41859-67-0 Fibrate C19H20ClNO4 361.83 2.12E-015 4.25 3.61
the process. All samples have been spiked with 100 µg/L standard isotopes, except TCPP
and TDCPP, the method was following the EPA method 8270, and the steps were as
follows:
Activation of sorbent: conditioned cartridge with 5.0 mL of MTBE and methanol into
Application of samples: loaded 50 mL into cartridge, original sample was 1.0 mL and
diluted to 50 mL;
Removal of interfering compounds: rinsed cartridge with 5.0 mL of waste into aqueous
Elution of analytes: washed cartridge with 5.0 mL of methanol and 5.0 mL of 10%
methanol with MTBE separately, and collected 5.0 mL fraction into sample tube using
The concentration of target OMPs were measured using Agilent Technology 1260 module
coupled with AB SCIEX QTRAP 5500 mass spectrometer (Applied Biosystems, Foster
City, CA). The LC unit consists of a degasser, a binary pump, an autosampler and an LC
The operation of the LC was separated into positive and negative mode for different OMPs,
and the detailed process was as following, and the MS information can be found in the
Appendix B.
Mobile phase for ESI +: 4 mM ammonium formate in water containing 0.1% formic acid
(A) and 4mM Ammonium formate in methanol containing 0.1% formic acid (B). Flow rate
of 800 uL/min as follows: 90% A held for 0.5 min, then stepped down to 50% at 0.51 min
and decreased linearly to 5% at 8 min. It is held at 5% for 6 mins, then stepped up to 90%
Mobile phase for ESI-: 2mM ammonium acetate in water (A) and 2mM Ammonium
acetate in methanol (B). Flow rate of 800 µL/min as follows: 90% A held for 0.5 min, then
stepped down to 60% at 0.51 min and decreased linearly to 5% at 8 min. It was held at 5%
till 11 min, then decreased linearly to 90% A in 3 min followed by a 4 min equilibration
The MLSS in the bioreactor was 3000 mg/L when the batch experiment began. In the batch
test, eight 250 ml Erlenmeyer flasks with 200 ml mixed liquor were run simultaneously at
20 ℃ for a week following the four treatments (reactor1, reactor2, reactor3, and reactor4)
shown in Table 3-2. The removal routes for OMPs in the activated sludge process were
considered to be biodegradtion (B), adsorption (A), volatilization (V), and hydrolysis (H).
For reactor1, a and b were duplicated reactors in which all of the removal routes will occur.
For reactor2 and reactor3, biodegradation was inhibited by NaN3 and high temperature
sterilization, respectively. For reactor4, activated sludge was not added, therefore, only
32
results and assumption, biodegradation and adsorption was based on the differences of
All reactors were covered with aluminum foil to void possible photolysis. OMPs were
spiked with a methanol stock solution mixture containing 1 mg/mL of each OMPs, and
final OMPs concentrations in the batch reactors were 100 µg/L. The high concentration in
the stock was planed to avoid increasing the COD by methanol stock. Synthetic wastewater
as described in Chapter 2, and activated sludge was taken just after the fill mode in the
bioreactors, therefore, COD was kept same as the bioreactor influent. Pure oxygen was
used for aeration with a pressure of 0.1 MPa, and DO in the batch reactors was around 80%
at 20 ℃. Stirring was supplied by magnetic stirrers at 350 rpm. Samples were taken from
the batch reactor at the following times: 0, 0.25, 0.5, 1, 2, 5, 12, 24, 36, 48, 72, 96, 120, 144,
168h, respectively. The collected samples will be first centrifuged at 4000 G for 10 minutes,
and then filtered through 0.45µm glass fiber filter. The filter was cleaned by M.Q. water
and injected air to squeeze out the liquid. The first 1 mL filtrate was discarded, and the
following was collected in a glass centrifuge tubes. All of the samples were carried out in
duplicate, and a control sample without sludge was also prepared under the same condition
to determine if there was a loss of OMPs. The whole design sketch is shown in Figure 3-1.
33
4000G 10 mins
+
0.45 µm glass fiber
filter
SPE
LC/MS/MS
R1:2000 mg/L sludge R2:2000 mg/L sludge R3:2000 mg/L R3:100µg/L OMPs
+ + autoclaved sludge +
100µg/L OMPs 100µg/L OMPs + 0.1% NaN3
+ 100µg/L OMPs
0.1% NaN3 +
0.1% NaN3
Design of reactor2 was applied to find the sorption equilibrium time. Samples were taken
at 0, 24, 48h, respectively. OMPs concentration was checked, and the equilibrium time was
determined.
Design of reactor2 was also applied in the determination of sorption isotherms after the
equilibrium time was determined (48h was set as the sorption equilibrium time). The
sorption isotherms were conducted using 1, 10, 50, and 100 ug/L OMPs. The injection
volumes of the OMPs were the same to keep same COD in each reactor, stocks with
different concentration were prepared, and the spiking volumes are shown in Table 3-3.
Three isotherms including the Freundlich, linear, and Langmuir isotherms were applied to
describe the sorption equilibrium. During the sorption experiments, OMPs concentration in
the liquid phase were used to determined their partitioning into the sludge as follows:
qeq
C
0 Ceq
(2.3)
M
35
Where qeq is the OMPs partition into the sludge, C0 is the initial concentration of OMPs,
Ceq is the residual OMPs concentration, and M is the concentration of the sludge.
Freundlich model:
Where qeq (mg/g) is the equilibrium amount OMPs that adsorbed onto the sludge, k f is
the Freundlich adsorption coefficient, Ceq is the equilibrium concentration of OMPs in the
liquid phase, and 1/n is the measurement of nonlinearity. Freundlich isotherm can be
1
log qeq log K f log Ceq (2.5)
n
Linear model:
Langmuir model:
QbCeq
qeq (2.7)
1 bCeq
36
Where Q is the binding strength, b is the maximum amount of the compound absorbed per
Ceq 1 Ceq
(2.8)
qeq Qb Q
Lagergren model which is a pseudo-first order rate equation was used to simulate the
adsorption kinetics:
K1 qeq q
dq
(2.9)
dt
K1
log qe qt log qe t (2.10)
2.303
Three kinetic models were applied to simulate the results for biodegradation, which were
Zero-order:
dC
k0 Ct C0 k0t (2.11)
dt
First-order:
dC
k1C Ct C0e k1t (2.12)
dt
Second-order:
37
dC
k2C 2 Ct C0 / 1 C0 k2t (2.13)
dt
K0 (µg L-1 h-1), K1 (h-1) and K2 (L µg-1 h-1) are the zero-order, first-order, second-
Due to the complexity of the sludge composition, the results for the removal of OMPs by
different groups varied greatly. It was difficult to separate biodegradation and bio-
In the control groups, no obvious changes of OMPs concentration was found, therefore, the
removal by hydrolysis and volatilization was ignored in the study. This result could be
explained by the low Henry’s constants, relative high molecular weight and polarized
functional groups (Table 3-1 and Appendix A); several other groups also found the same
conclusion that the removal by hydrolysis and volatilization could be ignored [39, 41-44].
Many studies had employed NaN3 to inhibit the aerobic sludge activities by paralyzing
would not work in the process [41, 42]. Li et al. [42] had proposed to use caffeine to exam
38
the inhibition ability. It was based that caffeine was easy to be degraded by microbes while
hard to be adsorbed by sludge. In their studies, caffeine was biodegraded completely within
10 hours without adsorption. Judging from our result (see Figure 3-2), caffeine was not
adsorbed but the degradation was also not obvious. This was due to the differences
between the sludge rather than the weak biodegradation ability, as other target OMPs like
bisphenol A and naproxen were found to be biodegradated well. Therefore, caffeine was
not a perfect indicator in our study, and we choose COD changes as a parameter in this
case. COD of the samples taken at 0h and 168h was measured, and the result is shown
in Table 3-4. The 58.6% removal in reactor1 indicated that biodegradation was occurring,
while there was not much change in other reactors. In reactor 3, the COD was much higher
than the others; this was because of the cell rupture during sterilization, which brought the
intracellular substances into the liquid phase. The results indicated that NaN3 could inhibit
the sludge in our study, as COD associated with the synthetic wastewater was easier
biodegraded by the activated sludge than OMPs, while there were not much COD changes
when NaN3 was spiked into the reactors. Therefore, the OMPs removal in reactor 2 was
Table 3-4. COD changes between 0h and 168h in the adsorption and
biodegradation experiment
Reactor 1 Reactor 2 Reactor 3 Reactor 4
0h 168h 0h 168h 0h 168h 0h 168h
COD (mg/L) 355 147 473 484 773 710 444 446
39
Adsorption
1.2 Biodegradation
Biodegradation+adsorption
1.0
0.6
0.4
0.2
0.0
0 20 40 60 80 100 120 140 160 180
Time (h)
3.5.1 Isotherms
It was found that many OMP like methylparaben, propylparaben, and atenolol could be
well adsorbed by activated sludge. However, it was hard to determine the isotherm due to
three reasons: OMPs like methylparaben and propylparaben were totally adsorbed within
few hours; therefore, the equilibrium sorption amount could not be decided. Other OMPs
like diclofenac, and sulfamethoxazole were continually been adsorbed by the sludge rather
than reaching equilibrium, therefore, the sorption equilibrium time we used was not
40
applicable in the case. Therefore, the sorption equilibrium curves will be not presented in
this study.
It was found that acesulfame, clofibric acid, dilatin, iopromide, primidone, sucralose, TCPP,
TDCPP and trimethoprim were not removed by aerobic activated sludge, part of results
corresponded to the result by other groups [45, 46]. It was easy to explain that there was
not much adsorption for those OMPs as they reflected negative or neutral charges, which
would be electrostatic repulsed, the result was in agreement with the Kd value in Table 3-1,
as Joss et al. [47] proposed that the adsorption could be ignored when Kd was lower than
300 L/kg SS. In addition, due to the high molecular weight as well as stable structures of
the OMPs, they were hard to be biodegraded by the sludge. TCPP and TDCPP were the
typical chemicals that were difficult to remove by aerobic activated sludge as chlorine was
high in their structures. Moreover, the bacteria culture may not be adapted to certain
Typical OMPs was selected to be presented for their adsorption properties in this section,
and the information for the other OMPs can be found in the appendix.
It was found that in reactor2 that the adsorption of atrazine, sulfamethoxazole, amitriptyline,
and TCEP kept being adsorbed by sludge. For atrazion, naproxen, and TCEP, the apparent
sorption equilibrium were reached at more than one days, shown in [42] which was not in
agreement with the Lagergren model, which usually uses 3-6 hours as the equilibrium time.
The same phenomenon was observed by Yu et al. [48]. The adsorption for
41
sulfamethoxazole was following a zero-order model, and Kp0 was 0.00265 mgL-1h-1 (R2:
0.999). In general, the zero-order kinetics were not applicable in adsorption processes as
the adsorption should reach equilibrium at a certain time with the desorption processes.
This phenomena could be explained that the sorption process of sulfamethoxazole was a
two-step process adsorption, in the first step, the adsorption was quick which was not
related to the sulfamethoxazole concentration; and then in the second step, while the
sorption points was taken, the sorption slowed down, and finally reach equilibrium. Further
1.2 Adsorption
A B
1.0
1.1
0.8
Adsorption (C/C0)
1.0
0.6
C/C0
Adsorption
Biodegradation 0.9
0.4 Biod+ads
0.8
0.2
0.0 0.7
0 20 40 60 80 100 120 140 160 180 0 20 40 60 80 100 120 140 160 180
Time (h) Time (h)
Figure 3-3. The removal of atrazine by aerobic activated sludge. A total removal, B adsorption curve
1.2
A B
Adsorption Adsorption (C/C0)
1.0
Biodegradation
Fitting curve
Biod+ads
1.0
0.8
0.8
0.6
C/C0
C/C0
0.6
0.4
0.4
0.2
0.0 0.2
0 20 40 60 80 100 120 140 160 180 0 50 100 150
Time (h) Time (h)
Figure 3-4. The removal of sulfamethoxazole by aerobic activated sludge. A total removal, B zero order
simulation curve
43
For OMPs like atenolol, methyparaben, and propylparaben, the complete adsorption
occurred within 8 hours. Zero order and first order kinetics was applied to simulate the
processes. Propylparaben was firstly fitted with the zero-order model, and the R2 was 0.416,
while R2 for first order model was 0.999, therefore, first-order adsorption model was
selected, as shown in Figure 3-5. The analysis of the other OMPs was done with the same
method, i.e. choosing the model with higher R2 value, and the results is shown in Table 3-5.
The strong adsorption of atenolol could be explained by its positive charge, as the sludge
exhibited a negative charge, the electrostatic attraction made the adsorption process
stronger than other routes. Methylparaben and propylparaben were also positively charged,
and their adsorption was found to be quicker than atenolol. The reason for this
phenomenon was their phenolic group, which may also enhance the adsorption process.
For the study of adsorption of OMPs by aerobic activated sludge, different inhibition
methods have been applied by different groups; among them, NaN3 and high temperature
sterilization were the most commonly used methods. The advantage of NaN3 is it will not
damage the cells, while its inhibition ability is effective for aerobic sludge. However, the
addition of NaN3 will isolate the mechanism for target compounds [49]. The advantage of
high temperature sterilization inhibition is it could guarantee the total inhibition of the
sludge, while it could break the cells which provide new sorption points, and change the
sludge charge as well. It was found that there were no more adsorption of atenolol and
atrazine after high temperature sterilization, noting that electrostatic attraction played main
role in the adsorption of atrazine, the phenomena could be explained by the change of
44
sludge charge. Methylparaben and propylparaben were not detected in the high temperature
sterilization experiment, which made it difficult for further analysis. For the other OMPs ,
adsorption (C/C0)
1.0
A 1.0 B
adsorption
0.8 Biodegradation
Bio+adsorp
adsorption (C/C0)
adsorption (C/C0)
Control
0.6
0.5
0.4
0.2
0.0 0.0
0 2 4 6 8 10 12 0 1 2
Time (h) Time (h)
Figure 3-5. The removal of propylparaben by aerobic activated sludge. A total removal, B first-order
simulation curve
For most target OMPs, biodegradation was the main removal route [41, 50, 51]. Zero-order
and first-order reaction were the main removal kinetics for the experiment. The result are
45
shown in Table 3-6 and Table 3-7. Benzafibrate, bisphenol A, diclofenac, gemgibrozil,
ibuprofen, caffeine, and DEET fit the zero-order biodegradation kinetics. Ibuprofen was
completely removal by both adsorption and biodegradation, Nakada et al. [52] found the
reaction (KBDS=0.88d-1, R2=0.955), and the constants we simulated was close as reported
naproxen was not high, which was in agreement with Nakada et al.[52]. The possible
explanation was the lower hydrophobicity (log Kow<3) of amide-type pharmaceuticals. Joss et al
[47] found that removal of selected pharmaceuticals was mainly due to biological transformation
result; they also proposed that when the sorption coefficient was below 300 L/kg, adsorption could
be ignored. Table 3-8 showed the removal comparison between the literature and our study, and it
was found that CAS was effective for the removal of certain OMPs, however, the result varies
significantly between different groups, and few literatures have reported the mechanisms behind the
meaningful not only in providing data, but also for the study of mechanisms of OMPs
removal.
A B Biodegradation (C/C0)
1.0 Bio+adsorp (C/C0)
1.0 Fitting curve
Relative concentration(C/C0)
0.8 0.8
Bio+adsorp (C/C0)
0.6
0.6
0.4
0.4
0.2
0.0
0 2 4 0 50 100 150
Time (h) Time (h)
Figure 3-6. Simulation of removal ibuprofen by aerobic activated sludge. A first-order removal kinetics
by sorption and biodegradation, B zero-order biodegradation kinetics
3.6 Summary
In this chapter, experiment was designed to study the removal mechanism of OMPs by
aerobic activated sludge. It was found that biodegradation and adsorption were the main
removal routes, while volatilization and hydrolysis could be ignored in the process. NaN3
proved to be an effective inhibitor for the aerobic bacteria, and the comparison between the
played an important role in the adsorption process. Target OMPs responded to the two
47
weak sorption and weak biodegradation by atrazine, DEET, diclofenac, gemfibrozil, TCEP,
and trimethoprim. Kinetic study showed that adsorption of atenolol, mathylparaben and
propylparaben well followed first-order model (R2: 0.939 to 0.999) with the rate constants
ranging from 0.519-7.092 h-1. For biodegradation kinetics, it was found that benzafibrate,
model (K0:1.15E-4 to 0.0142 µg/Lh-1, R2: 0.991 to 0.999), while TCEP, naproxen,
Table 3-8. OMPs removal rate comparison between literature and the study
OMPs Literature Batch test
Biodegradation method Total method Biodegradation Total removal
removal rate removal rate removal rate rate
% % % %
Acesulfame n.d. n.d. n.d. n.d. n.d. n.d.
Amitriptyline n.d. n.d. n.d. n.d. 61 88
Atenolol n.d. n.d. 33.5[53], SBR, lab scale n.d. n.d.
36[53]
Atrazine n.d. n.d. ND 24
Bezafibrate 100[54] Batch 100[54] Batch 55 63
Bisphenol A 62 97
Caffeine 68 ± 10 [55] Batch 92.2[56] Batch 50 50
Carbamazepine n.d. n.d. <20[57] SBR, lab scale <5 n.d.
53.5[56] Batch
Clofibric acid 26–30[58] Lab 71.8[59] MBR, lab scale n.d. n.d.
columns 16.7, Lab columns
48.3[60]
DEET n.d. n.d. 19.2– WWTP, full 11 24
46.2[52] scale
Diclofenac 34–38[58] Lab n.d. n.d. 51 66
columns
30 Batch
Dilantin n.d. n.d. 76.8[56] Batch n.d. n.d.
Diphenhydramine n.d. n.d. n.d. n.d. 68 77
Gemfibrozil >99[48] Batch 89.6[59] MBR, lab scale 10 40
98.9[56] Batch
Ibuprofen n.d. n.d. <20[57] SBR, lab scale 63 89
96.3[60] Lab columns
98.9[56] Batch
Iopromide 86-97[61] Batch 100[60], Lab columns n.d. n.d.
85[62] 48.8[60]
33.0[56] Batch
Methyparaben n.d. n.d. n.d. n.d. n.d. ~100
Naproxen 60[54],80[48] Batch 75.9[60] Lab columns 53 93
97.8[56] Batch
Oxybenzone n.d. n.d. 41[63] MBR, pilot 43 49
scale
98.9[56] Batch
Primidone n.d. n.d. n.d. n.d. n.d. n.d.
Propylparaben n.d. n.d. n.d. n.d. n.d. ~100
Sucralose n.d. n.d. n.d. n.d. n.d. n.d.
Sulfamethoxazole n.d. n.d. 70[63] MBR, pilot 77 ~99
and lab scale
~100[64] SBR, lab scale
77.3[56] Batch
TCEP n.d. n.d. n.d. n.d. <5 38
TCPP n.d. n.d. n.d. n.d. n.d. n.d.
TDCPP n.d. n.d. n.d. n.d. n.d. n.d.
Trimethoprim n.d. n.d. 23.8[56] Batch n.d. <2
49
50
4.1 Introduction
For many of the target OMPs, they do not always exist in wastewater. Some of the OMPs
changes of removal ability of those OMPs in non-acclimated and acclimated sludge. It was
found that for some OMPs, the removal had been enhanced during the adaptation, which
showed an OMPs concentration decrease in the effluent water; for most of the OMPs, the
OMPS concentration stayed the same, which could be explained by quick adaptation or the
culturing time is too short for obvious adaptation. A noticeable result was found that some
Right after the OMPs removal mechanism experiment, 10 µg/L OMPs stock was spiked
into the synthetic wastewater every day, and the OMPs concentration was monitored. After
two months of operation, the sludge was taken, and the removal mechanism experiment
was repeated again to check if the removal rate constants were changed.
Materials were the same as described in Chapter 2 and Chapter 3, the operation of SBR
reactor was described in Chapter 2, and the OMPs concentration measure methods and
After two months of monitoring, the effluent OMPs concentration data were collected, and
Biodegradation played the most important role in the enhancement of the removal ability.
Bisphenol A, ibuprofen and naproxen showed quick adaptation (Figure 4-1and Figure 4-2).
The batch test results proved this phenomenon, and it was found the removal rate was
increased. Diclofenac showed slower adaptation and the removal adaptation of iopromide
was even lower, see Figure 4-3. As described in Chapter 3, diclofenac showed weak
compound. The results indicated that the initial removal ability was an important parameter
Bisphenol A
14
A 1.0
B non-acclimated removal curve
acclimated removal curve
10
Bisphenol A (ng/mL)
0.5
6
0.0
0
0 20 40 60 80 100 120 140 160 180
0 10 20 30 40 50
time (h)
Time (d)
Figure 4-1. (A) removal trend of bisphenol A. (B)removal of bisphenol A by non-acclimated and
acclimated activated sludge
Ibuprofen
4
Ibuprofen (ng/mL)
0.5
2
0 0.0
0 10 20 30 40 50 60 0 20 40 60 80 100 120 140 160
Time (d) time (h)
Figure 4-2. (A) removal trend of ibuprofen. (B)removal of ibuprofen by non-acclimated and acclimated
activated sludge
53
Diclofenac Iopromide
10 10
A B
8 8
Diclofenac (ng/mL)
Iopromide (ng/mL)
6 6
4 4
2 2
0 0
0 10 20 30 40 50 0 10 20 30 40 50
Time (d) Time (d)
adaptation during the two months operation, see Figure 4-4 and appendix. Note that
atenolol and trimethoprim were positively charged, and were well adsorbed by the
negatively charged sludge, they showed a low concentration in the effluent at the beginning.
The atenolol concentration kept low for about 30 days and then began to increase; this was
due to the large amount of sorption points for atenolol. Adsorption played the most
important role in the removal process (completely absorbed within 10 hours at 100 µg/L),
after the adsorption reached equilibrium, i.e., sorption points were fully occupied, no more
concentration showed the same trend as atenolol concentration, however, the adsorption
process was not as fast as that of atenolol, therefore, the initial concentration was not as
low as atenolol. The equilibrium concentration reached the influent concentration, which
Trimethoprim
7 Atenolol 10
A 9
B
6
8
5 7
Trimethoprim (ng/mL)
Atenolol (ng/mL)
6
4
5
3
4
2 3
2
1
1
0 0
0 10 20 30 40 50 60 0 10 20 30 40 50 60
The stable trend could be classified into two situations. It was obvious that for the difficult
to remove compounds, the effluent concentration would stay the same as the influent
concentration during the operation. clofibric acid ,gemfibrozil , primidone ,and TCPP fit
this situation, the batch removal test also proved the results, see Figure 4-5. Although
removal in the operation, this was probably due to the bacteria culture changes, in which
the bacteria that could remove the compounds was eliminated in the competition,
55
see Figure 4-5. In another situation, the effluent concentration was lower than the influent
concentration, see Figure 4-6. This could be explained by either a quick adaptation, which
made the decrease trend not obvious, or the removal ability stayed the same during the
operation. What’s more, it was easy to draw the conclusion that the lower the effluent
concentration, the stronger the removal ability. Therefore, the removal ability order by
and sulfamethoxazole was proven in Chapter 3. The strong removal of DEET may be
9
10
8
Clofibric Acid (ng/mL)
7
Bezafibrate (ng/mL)
8
6
6 5
4
4
3
2
2
A 1
B
0 0
0 10 20 30 40 50 60 0 10 20 30 40 50
Time (d) Time (d)
Figure 4-5. Removal trend of clofibric acid (A) and benzefibrate (B)
56
DEET
Clofibric Acid
14 Oxybenzone
Sulfamethoxazole
12 TCEP
Dilantin
10
DEET (mg/L)
0
0 10 20 30 40
Time (d)
Figure 4-6. Removal trend of DEET, clofibric acid, oxybenzone, sulfamethoxazole, TCEP and dilatin
4.4 Summary
The adaptation trend was examined in different ways, typically: decreasing, increasing,
stable. For the decrease situation, the reason was a positive adaptation, which enhanced the
sludge removal ability, and the adaptation time was corresponded to the initial removal
ability. What is more, the increasing trend was mainly caused by strong absorption by the
sludge, once the sorption points were occupied; the compound concentration began to
increase. Finally, the stable trend occurred whether no removal or quick adaptation removal.
The lower the stable concentration was, the stronger the removal ability was.
57
5.1 Introduction
separation, has got a rapid growth in both domestic and industrial wastewater treatment and
reuse in recent years due to its significant advantages over conventional activated sludge
process such as better effluent quality and less footprint.However, the microfiltration (MF)
and ultrafiltration (UF) membrane widely used in membrane bioreactor cannot reject OMPs
due to their much higher pore size or molecular weight cut-off (MWCO) than OMPs
(see Figure 5-1). In order to enhance OMPs removal, nanofiltration (NF) is a promising
alternative to MF/UF in MBR as it can effectively reject the OMPs with molecular weight
over its MWCO,. A NF module was connected to the former SBR reactor to form an
aerobic nanofiltration membrane bioreactor for one month operation in this study. The
rejection of OMPs by NF would increase their retention time in the bioreactor, . Based on
the concept of hydraulic retention time (HRT) and sludge retention time (SRT), the concept
of compound retention time (CRT) for OMPs in the aerobic nanofiltration membrane
bioreactor was proposed and analyzed in this chapter.The overall performance of OMPs
Membrane Cell System, and all the other materials and devices were described in Chapter 2
and Chapter 3.
5.1.2 Methods
The experiment design was shown in Figure 5-2. The system was combined with a
bioreactor and a membrane filtration system. As the bioreactor was running in SBR mode,
59
the hybrid system was named nanofiltration sequencing batch membrane bioreactor (NF-
SB-MBR). The operation of the SBR was the same as descripted in Chapter 2, 4 L
synthetic wastewater with 10 µg/L OMPs was added into the reactor daily. The effluent
was collected daily and its OMPs concentration was treated as OMPs concentration in the
bioreactor, Cin (µg/L). And then, the collected effluent (4 L) from the bioreactor was
filtered by Dow NF90 membrane in Sterlitech crossflow membrane cell system. Trans-
membrane pressure (TMP) was kept at 10 bar. Permeate was collected in a tank on an
electronic balance connected to a computer for recording permeate weight with time
continuously, and thus permeate flux was calculated. The filtration process was stopped
when 2 L permeate was obtained. The NF permeate was sampled, and its OMPs
collected and used to make the synthetic wastewater. The actual daily treated wastewater
volume was 2 L, and thus the HRT was 1 d. The each OMP dosage was 40 µg/d and the
same as the previous SBR operation, which resulted in the average influent concentration
Valve
Influent
P
Influent
Pressure gage
NF
Tank
Concentrate
SBR
Figure 5-2. Schematic diagram of aerobic nanofiltration membrane bioreactor (The bioreactor was
running in SBR mode, SBR effluent was sampled as Cin and filtered by NF membrane in a constant-
pressure crossflow membrane cell , the permeate was collect and sampled as the final effluent (Cp),
and the concentrate was back to the SBR.)
CinVR
CRT (5.1)
C p Q p Cin Qs
concentration (µg/L), Qs is sludge discharge flow (L/d), Cin is the average OMP
concentration in reactor (µg/L). All the sludge was rejected by the membrane, and no waste
sludge was discharged during the operation. Therefore, (5.1) could be simplified as follow:
CinVR Cin
CRT HRT (5.2)
C p Qp C p
61
a 1 C p / Cin (5.3)
R 1 C p / 20 (5.4)
For OMPs biodegradated by the first-order kinetics, the degradation rate R1 is defined as
(5.5)
dCin
R1 K1Cin (5.5)
dt
When CRT reached steady state, i.e. Cp and Cin were stable, we can assume R1 is stable in
R1
1
HRT
C f C p (5.6)
From (5.5) and (5.6), we can deduce the first-order reaction rate under steady state,
K1
1
HRT Cin
C f C p (5.7)
R0
dCin
dt
K0
1
HRT
C f C p (5.8)
62
For some compounds including acesulfame, caffeine, clofibric acid, diclofenac, dilatin,
gemfibrozil, iopromide and sulcralose, there were 1-3 times higher concentrations in the
bioreactor than in the feed, indicating their significant accumulation in the bioreactor due to
the effective rejection by NF membrane. These OMPs were found nearly no or very little
removal in previous study. As an example, the concentration in the bioreactor (Cin) and the
permeate (Cp), the NF rejection and the overall removal, and CRT changes of acesulfame
were shown in Figure 5-3. The information of other compounds could be found in
Appendix E. It was found CRT was highly dependent on NF rejection rate. The higher the
rejection rate was the higher CRT was (Figure 5-4). The permeate concentration of
bisphenol A at stable time (20 days later) was lower than the influent concentration. Take
consideration of the high removal by activated sludge, it can be predicted that the removal
bisphenol A was enhance during the operation (Figure 5-5). Amtripline was a readily
removable compound, and the effluent concentration kept stable and low, see Figure 5-6.
The high CRT (around 200d), which was caused by high rejection rate may contribute to
the removal process. Figure 5-8 shows the permeate concentration of target OMPs, most of
them are under 4 µg/L except acesulfame, caffeine, clofibric acid, diclofenac, dilatin,
gemfibrozil, iopromide and sulcralose, of which the concentration in the bioreactor were
increased. The overall removal rate was over 80% for most of the OMPs. The result
indicates the NF-MBR is promising for removal of the selected compounds. Naproxen was
well biodegradable and difficult adsorbed compound, and the biodegradation followed
first-order reaction kinetic (K1=0.0101 h-1). The operational parameters of naproxen in NF-
SB-MBR are shown in Figure 5-7. CRT was stable after 20 days, using (5.7) the first-order
63
reaction constant is 0.0704 h-1, which is much higher than the starting value. It indicates the
biodegradation was greatly enhanced during the operation. For OMPs followed zero-order
reaction, like bisphenol A and benzafibrate, CRT kept decreasing, therefore, zero-order
reaction constant was not calculated in this study. The decreasing of CRT indicates OMPs
concentration in the bioreactor was greatly decrease, as the permeate concentration was
stable.
Cin (µg/L) 60
70 Cp (µg/L) 55
CRT (d) 50
60
OMPs concentration (µg/L)
45
50 40
35
CRT (d)
40
30
30 25
20
20 15
10
10
5
0 0
0 5 10 15 20 25 30 35
time (d)
100
30 CRT
rejection rate
95
25
15 85
10
80
5
75
0
0 5 10 15 20 25 30 35
time (d)
Cin (µg/L)
Cp (µg/L)
CRT (d) 40
8
35
7
OMPs concentration (µg/L)
30
6
25
5
CRT (d)
20
4
15
3
10
2
5
1
0
0
0 5 10 15 20 25 30 35
time (d)
5 600
Cin (µg/L)
Cp (µg/L)
4 CRT (d) 500
300
2
200
1
100
0
0
0 5 10 15 20 25 30
time (d)
Cin (µg/L)
20 60
Cp (µg/L)
18 CRT (d) 55
50
Naproxen concentration(µg/L)
16
45
14
40
12 35
CRT (d)
10 30
8 25
20
6
15
4
10
2 5
0 0
10 15 20 25 30
time (d)
20
18
Effluent concentration(µg/L)
16
14
12
10
8
6
4
2
0
Acesulfame
Amitriptyline
Atenolol
Atrazine
Benzophenone
Bezafibrate
Bisphenol A
Caffeine
Carbamazepine
Clofibric Acid
DEET
Diclofenac
PS
Dilantin
Diphenhydramine
OM
Fluoxetine
Gemfibrozil
Ibuprofen
Iopromide
Naproxen
Oxybenzone
Primidone
Sucralose
Sulfamethoxazole
TCEP
TCPP
TDCPP
Trimethoprim
0 5 10 15 20 25 30
time (d)
6.1 Conclusions
A typical aerobic non-acclimated activated sludge was cultured by SBR mode for 6 months.
OMPs removal mechanisms were studied using the sludge by a batch test. Hydrolysis and
volatilization could be ignored in the removal process, but adsorption and biodegradation
were the main routes. For compounds like methylparaben, adsorption was the only removal
mechanism due the strong electrostatic attraction, while for compounds like bisphenol A,
mechanism were explained, and proper models were used to simulate the kinetics. What is
more, NaN3 was an effective inhibitor for aerobic sludge. The inhibition between NaN3 and
high temperature sterilization was compared, and NaN3 inhibition was suggested to be a
The removal of OMPs by non-acclimated and acclimated activated sludge was compared. It
was found that the bioreactor effluent OMPs concentration showed three trends: decreasing,
increasing, and stable. The decreasing trend suggested an adaptation of the sludge, the
stronger initial removal, the quicker it could reach new equilibrium. The increasing trend
was mainly caused by strong adsorption, in which the OMPs concentration was very low
before the sorption points were fully occupied. The stable trend could be classified into two
situations: no removal and quick removal. For the latter situation, the stable OMPs
concentration suggested the removal ability: low concentration indicated strong removal.
68
The stable concentrations were compared, and the result corresponded to the batch test
results.
A novel NF-SB-MBR operation was proposed and operated. Several OMPs concentration
in the bioreactor were found to increase intensively, which suggested good rejection by NF
membrane. CRT was related to rejection rate, high rejection rate indicated long CRT.
6.2 Suggestions
In the adaptation test, molecular biology analysis is needed. The results will give
A batch test of the removal kinetic after the NF-SB-MBR was conducted to find if there is
The adsorption mechanism was not yet fully understood. Usually, the adsorption was
related to extracellular polymeric substance (EPS), yet few studies have studied the role of
EPS in adsorption in CAS, and an EPS extraction method need to be developed and
evaluated.
References
16. Altenburger, R., et al., Predictability of the toxicity of multiple chemical mixtures to
Vibrio fischeri: Mixtures composed of similarly acting chemicals. Environmental
Toxicology and Chemistry, 2000. 19(9): p. 2341-2347.
17. Jones, O.A.H., et al., Questioning the Excessive Use of Advanced Treatment to
Remove Organic Micropollutants from Wastewater. Environmental Science &
Technology, 2007. 41(14): p. 5085-5089.
18. Sedlak, D.L.; Available from: http://www.ce.berkeley.edu/~sedlak/.
19. EPA, U.S.; Available from: http://www.epa.gov/ppcp/.
20. Ternes, T., Pharmaceutical pollutants. Biofutur, 2001. 2001(216): p. 44-46.
21. Ternes, T., M. Bonerz, and T. Schmidt, Determination of neutral pharmaceuticals in
wastewater and rivers by liquid chromatography-electrospray tandem mass
spectrometry. Journal of Chromatography A, 2001. 938(1-2): p. 175-185.
22. Kolpin, D.W., et al., Pharmaceuticals, hormones, and other organic wastewater
contaminants in US streams, 1999-2000: A national reconnaissance. Environmental
Science & Technology, 2002. 36(6): p. 1202-1211.
23. Alder, L., et al., Estimation of measurement uncertainty in pesticide residue
analysis. Journal of Aoac International, 2001. 84(5): p. 1569-1578.
24. Golet, E.M., et al., Trace determination of fluoroquinolone antibacterial agents in
solid-phase extraction urban wastewater by and liquid chromatography with
fluorescence detection. Analytical Chemistry, 2001. 73(15): p. 3632-3638.
25. Hirsch, R., et al., Determination of antibiotics in different water compartments via
liquid chromatography–electrospray tandem mass spectrometry. Journal of
Chromatography A, 1998. 815(2): p. 213-223.
26. Lindsey, M.E., M. Meyer, and E.M. Thurman, Analysis of Trace Levels of
Sulfonamide and Tetracycline Antimicrobials in Groundwater and Surface Water
Using Solid-Phase Extraction and Liquid Chromatography/Mass Spectrometry.
Analytical Chemistry, 2001. 73(19): p. 4640-4646.
27. Hartmann, A., et al., Identification of fluoroquinolone antibiotics as the main
source of umuC genotoxicity in native hospital wastewater. Environmental
Toxicology and Chemistry, 1998. 17(3): p. 377-382.
28. Schantz, S.L. and J.J. Widholm, Cognitive effects of endocrine-disrupting chemicals
in animals. Environmental Health Perspectives, 2001. 109(12): p. 1197-206.
29. Snyder, S.A., et al., Pharmaceuticals, personal care products, and endocrine
disruptors in water: Implications for the water industry. Environmental Engineering
Science, 2003. 20(5): p. 449-469.
30. I. Nopens, C.C., P.A. Vanrolleghem, Stability analysis of a synthetic municipal
wastewater, 2001: Department of Applied Mathematics, Biometrics and Process
Control, University Gent.
31. Carballa, M., et al., Comparison between the conventional anaerobic digestion of
sewage sludge and its combination with a chemical or thermal pre-treatment
concerning the removal of pharmaceuticals and personal care products. Water
Science and Technology, 2006. 53(8): p. 109-117.
32. Sipma, J., et al., Comparison of removal of pharmaceuticals in MBR and activated
sludge systems. Desalination, 2010. 250(2): p. 653-659.
71
33. Giger, W., et al., Occurrence and fate of antibiotics as trace contaminants in
wastewaters, sewage sludges, and surface waters. Chimia, 2003. 57(9): p. 485-491.
34. Ducey, S.B. and A. Sapkota, Presence of Pharmaceuticals and Personal Care
Products in the Environment - A Concern for Human Health? Contaminants of
Emerging Concern in the Environment: Ecological and Human Health
Considerations, 2010. 1048: p. 345-365.
35. Boxall, A.B.A., et al., Pharmaceuticals and Personal Care Products in the
Environment: What Are the Big Questions? Environmental Health Perspectives,
2012. 120(9): p. 1221-1229.
36. Verstraeten, I.M., T. Heberer, and T. Scheytt, Occurrence, characteristics, transport,
and fate of pesticides, pharmaceuticals, industrial products, and personal care
products at riverbank filtration sites. Riverbank Filtration: Improving Source-Water
Quality, 2002. 43: p. 175-227.
37. Ternes, T.A., Occurrence of drugs in German sewage treatment plants and rivers.
Water Research, 1998. 32(11): p. 3245-3260.
38. Qiao, T.J., et al., Occurrence and fate of pharmaceuticals and personal care
products in drinking water in southern China. Journal of Environmental Monitoring,
2011. 13(11): p. 3097-3103.
39. Stevens-Garmon, J., et al., Sorption of emerging trace organic compounds onto
wastewater sludge solids. Water Research, 2011. 45(11): p. 3417-3426.
40. Seyhi, B., et al., Modeling of sorption of bisphenol A in sludge obtained from a
membrane bioreactor process. Chemical Engineering Journal, 2011. 172(1): p. 61-
67.
41. Yu, T.H., et al., Biodegradation and bio-sorption of antibiotics and non-steroidal
anti-inflammatory drugs using immobilized cell process. Chemosphere, 2011. 84(9):
p. 1216-1222.
42. Li, B. and T. Zhang, Biodegradation and Adsorption of Antibiotics in the Activated
Sludge Process. Environmental Science & Technology, 2010. 44(9): p. 3468-3473.
43. Prado, N., J. Ochoa, and A. Amrane, Biodegradation and biosorption of tetracycline
and tylosin antibiotics in activated sludge system. Process Biochemistry, 2009.
44(11): p. 1302-1306.
44. Wu, C.X., A.L. Spongberg, and J.D. Witter, Sorption and biodegradation of selected
antibiotics in biosolids. Journal of Environmental Science and Health Part a-
Toxic/Hazardous Substances & Environmental Engineering, 2009. 44(5): p. 454-461.
45. Gobel, A., et al., Fate of sulfonamides, macrolides, and trimethoprim in different
wastewater treatment technologies. Science of the Total Environment, 2007.
372(2-3): p. 361-371.
46. Guo, Y.C. and S.W. Krasner, Occurrence of Primidone, Carbamazepine, Caffeine,
and Precursors for N-Nitrosodimethylamine in Drinking Water Sources Impacted by
Wastewater. Journal of the American Water Resources Association, 2009. 45(1): p.
58-67.
47. Joss, A., et al., Removal of pharmaceuticals and fragrances in biological
wastewater treatment. Water Research, 2005. 39(14): p. 3139-3152.
72
48. Yu, J.T., E.J. Bouwer, and M. Coelhan, Occurrence and biodegradability studies of
selected pharmaceuticals and personal care products in sewage effluent.
Agricultural Water Management, 2006. 86(1–2): p. 72-80.
49. Bailey, D.N. and P.I. Jatlow, Chemical-Analysis of Massive Crystalluria Following
Primidone Overdose. American Journal of Clinical Pathology, 1972. 58(5): p. 583-&.
50. Abegglen, C., et al., The fate of selected micropollutants in a single-house MBR.
Water Research, 2009. 43(7): p. 2036-2046.
51. Batt, A.L., S. Kim, and D.S. Aga, Comparison of the occurrence of antibiotics in four
full-scale wastewater treatment plants with varying designs and operations.
Chemosphere, 2007. 68(3): p. 428-435.
52. Nakada, N., et al., Pharmaceutical chemicals and endocrine disrupters in municipal
wastewater in Tokyo and their removal during activated sludge treatment. Water
Research, 2006. 40(17): p. 3297-3303.
53. Carucci, A., G. Cappai, and M. Piredda, Biodegradability and Toxicity of
Pharmaceuticals in Biological Wastewater Treatment Plants. Journal of
Environmental Science and Health, Part A, 2006. 41(9): p. 1831-1842.
54. Quintana, J.B., S. Weiss, and T. Reemtsma, Pathways and metabolites of microbial
degradation of selected acidic pharmaceutical and their occurrence in municipal
wastewater treated by a membrane bioreactor. Water Research, 2005. 39(12): p.
2654-2664.
55. Bradley, P.M., et al., Biotransformation of caffeine, cotinine, and nicotine in stream
sediments: Implications for use as wastewater indicators. Environmental
Toxicology and Chemistry, 2007. 26(6): p. 1116-1121.
56. Snyder, S.A., et al., Biological and Physical Attenuation of Endocrine Disruptors and
Pharmaceuticals: Implications for Water Reuse. Ground Water Monitoring &
Remediation, 2004. 24(2): p. 108-118.
57. Clara, M., et al., Removal of selected pharmaceuticals, fragrances and endocrine
disrupting compounds in a membrane bioreactor and conventional wastewater
treatment plants. Water Research, 2005. 39(19): p. 4797-4807.
58. Zwiener, C. and F.H. Frimmel, Short-term tests with a pilot sewage plant and
biofilm reactors for the biological degradation of the pharmaceutical compounds
clofibric acid, ibuprofen, and diclofenac. Science of the Total Environment, 2003.
309(1–3): p. 201-211.
59. Tauxe-Wuersch, A., et al., Occurrence of several acidic drugs in sewage treatment
plants in Switzerland and risk assessment. Water Research, 2005. 39(9): p. 1761-
1772.
60. Kümmerer, K. and A. Al-Ahmad, Biodegradability of the Anti-tumour Agents 5-
Fluorouracil, Cytarabine, and Gemcitabine: Impact of the Chemical Structure and
Synergistic Toxicity with Hospital Effluent. Acta hydrochimica et hydrobiologica,
1997. 25(4): p. 166-172.
61. Batt, A.L., S. Kim, and D.S. Aga, Enhanced Biodegradation of Iopromide and
Trimethoprim in Nitrifying Activated Sludge†. Environmental Science & Technology,
2006. 40(23): p. 7367-7373.
73
62. Kalsch, W., Biodegradation of the iodinated X-ray contrast media diatrizoate and
iopromide. Science of the Total Environment, 1999. 225(1–2): p. 143-153.
63. Kim, S.D., et al., Occurrence and removal of pharmaceuticals and endocrine
disruptors in South Korean surface, drinking, and waste waters. Water Research,
2007. 41(5): p. 1013-1021.
64. Drillia, P., et al., On the occasional biodegradation of pharmaceuticals in the
activated sludge process: The example of the antibiotic sulfamethoxazole. Journal
of Hazardous Materials, 2005. 122(3): p. 259-265.
65. Simon Judd, C.J., The MBR Book. Second edition ed2011, UK.
74
Acesulfame Ibuprofen
Amitriptyline Iopromide
Atenolol Methyparaben
Atrazine Naproxen
Bezafibrate Oxybenzone
Bisphenol A Primidone
Caffeine Propylparaben
75
Carbamazepine Sucralose
DEET TCEP
Diclofenac TCPP
Dilantin TDCPP
Diphenhydramine Trimethoprim
Gemfibrozil
76
Appendix B: MS parameter
ESI+ mode
ESI- mode
1.5
Adsorption
Biodegradation
1.0 Biod+ads
1.0
Adsorption
Biodegradation
C/C0
C/C0
Biod+ads
0.5
0.5
0.0 0.0
0 20 40 60 80 0 20 40 60 80 100 120 140 160 180
Time (h) Time (h)
Adsorption
1.0
Fitting curve 1.0
0.8
0.8
0.6
0.6
C/C0
C/C0
Adsorption
Biodegradation
0.4 0.4 Biod+ads
0.2 0.2
0.0 0.0
0 2 4 6 8 10 12 0 20 40 60 80 100 120 140 160 180
Time (h) Time (h)
adsorption simulation curve of atenolol by aerobic removal curves of atrazine by aerobic activated sludge
activated sludge
79
Adsorption
1.6
Biodegradation 1.0
1.4
Bio+adsorp Biodegradation (C/C0)
Fitting curve
Ralative concentration(C/C0)
Relative concentrtion(C/C0)
1.2
1.0 0.8
0.8
0.6
0.6
0.4
0.2
0.0
0 20 40 60 80 100 120 140 160 180 0 50 100 150
Time (h) Time (h)
Biodegradation (C/C0)
1.2 Adsorption
1.0
Biodegradation
1.0
Bio+adsorp
Relative concentration(C/C0)
Biodegradation (C/C0)
0.8 0.8
0.6
0.6
0.4
0.2
0.4
0.0
0 10 20 30 40 50
0 20 40
Time (h)
Time (h)
Adsorption
Biodegradation 1.1 Adsorption
1.2
Biod+ads 1.0
Biodegradation
1.1 Bio+adsorp
0.9
1.0
Relative concentration(C/C0)
0.8
0.9
0.8 0.7
0.7 0.6
C/C0
0.6 0.5
0.5 0.4
0.4 0.3
0.3 0.2
0.2 0.1
0.1 0.0
0.0
0 20 40 60 80 100 120 140 160 180 0 20 40 60 80 100 120 140 160 180
removal curves of caffeine by aerobic activated sludge removal curves of diclofenac by aerobic activated sludge
Biodegradation (C/C0)
1.0 Adsorption(C/C0) 1.0 Fitting curve
Fitting curve 0.9
0.9
0.8
Relative concentration(C/C0)
Relative concentration(C/C0)
0.8
0.7 0.7
0.6 0.6
0.5 0.5
0.4 0.4
0.3 0.3
0.2 0.2
0.1 0.1
0.0 0.0
0 50 100 150 0 50 100 150
Time (h) Time (h)
Adsorption
1.1 Biodegradation
Biod+ads Biodegradation (C/C0)
1.0
1.0
Fitting curve
0.9
0.8
0.7 0.8
0.6
C/C0
C/C0
0.5
0.4 0.6
0.3
0.2
0.1 0.4
0.0
0 10 20 30 40 50 60 70 80 0 20 40 60
Time (h) Time (h)
0.8
0.8
0.6
0.6
0.4 0.4
0.2
0.2
0.0
0.0
0 20 40 60 80 100 120 140 160 180 0 50 100 150
Time (h) Time (h)
removal curves of ibuprofen by aerobic activated biodegradation simulation curve of ibuprofen by aerobic
adsorption (C/C0)
Adsorption
0.5 Biodegradation
Bio+adsorp 0.5
0.0
0.0
0 10 20 30 40 50 0 2 4 6 8 10 12
Time (h) Time (h)
Adsorption
1.0 Biodegradation 1.0 Biodegradation (C/C0)
Bio+adsorp Fitting curve
Relative concentration(C/C0)
0.8
0.8
0.6
0.6
C/C0
0.4
0.4
0.2
0.0 0.2
0 20 40 60 80 100 120 140 160 180 0 20 40 60 80 100 120 140 160 180
Time (h) Time (h)
removal curves of naproxen by aerobic activated biodegradation simulation curve of naproxen by aerobic
1.2
Biodegradation (C/C0)
1.0
1.0
Biodegradation (C/C0)
0.8
0.8
0.6
C/C0
0.4 0.6
Adsorption
Biodegradation
0.2
Biod+ads
0.4
0.0
0 20 40 60 80 100 120 140 160 180 0 50 100 150
adsorption (C/C0)
1.0
1.0
Relative concentration(C/C0)
adsorption (C/C0)
Adsorption
0.5 Biodegradation 0.5
Bio+adsorp
0.0
0.0
0 20 40 0 1 2
Time (h) Time (h)
removal curves of propylparaben by aerobic activated adsorption simulation curve of propylparaben by aerobic
1.2 Adsorption
1.0 Adsorption (C/C0)
Biodegradation
Fitting curve
Biod+ads
1.0
0.8
0.8
0.6
C/C0
C/C0
0.6
0.4
0.4
0.2
0.0 0.2
0 20 40 60 80 100 120 140 160 180 0 50 100 150
Time (h) Time (h)
1.0
Biodegradation (C/C0) 1.0
Fitting curve
0.8
0.8
0.6
C/C0
C/C0
Adsorption
0.6 0.4 Biodegradation
Biod+ads
0.2
0.4
0.0
0 50 100 150 0 20 40 60 80 100 120 140 160 180
Time (h) Time (h)
biodegradation simulation curve of sulfamethoxazole removal curves of TCEP by aerobic activated sludge by
1.0
Adsorption (C/C0)
Fitting curve
0.9
0.8
C/C0
0.7
0.6
0.5
0 50 100 150
Time (h)
activated sludge
86
Acesulfame
14 7 Atenolol
13
6
12
11 5
Acesulfame (ng/mL)
Atenolol (ng/mL)
10
4
9
8 3
7
2
6
5
1
4
3 0
0 10 20 30 40 50 60 0 10 20 30 40 50 60
Removal trend of acesulfame in aerobic activated Removal trend of atenolol in aerobic activated sludge
sludge bioreactor
bioreactor
Atrazine
10 20 Benzophenone
18
16
8
Benzophenone (ng/mL)
14
Atrazine (ng/mL)
12
6 10
6
4
4
2 0
0 10 20 30 40 50 60 0 10 20 30 40 50 60
Time (d) Time (d)
Removal trend of atrazine in aerobic activated sludge Removal trend of benzophenone in aerobic activated
Bezafibrate Bisphenol A
10 14
9
12
8
7 10
Bisphenol A (ng/mL)
Bezafibrate (ng/mL)
6
8
5
6
4
3 4
2
2
1
0 0
0 10 20 30 40 50 0 10 20 30 40 50
Time (d) Time (d)
Removal trend of benzafibrate in aerobic activated Removal trend of bisphenol Ain aerobic activated
Caffeine Carbamazepine
14
10
12
Carbamazepine (ng/mL)
8
10
Caffeine (ng/mL)
6 8
6
4
2
2
0 0
0 10 20 30 40 50 0 10 20 30 40 50 60
Removal trend of caffeine in aerobic activated sludge Removal trend of carbamazepin in aerobic activated
10
8
Clofibric Acid (ng/mL)
DEET (ng/mL)
6
4
4
2
2
0 0
0 10 20 30 40 50 60 0 10 20 30 40 50 60
Time (d) Time (d)
Removal trend of clofibric acid in aerobic activated Removal trend of DEET in aerobic activated sludge
Diclofenac Dilantin
10
10
8
8
Diclofenac (ng/mL)
Dilantin (ng/mL)
6
6
4 4
2 2
0 0
0 10 20 30 40 50 0 10 20 30 40 50
Time (d) Time (d)
Removal trend of diclofenac in aerobic activated Removal trend of dilatin in aerobic activated sludge
Diphenhydramine Fluoxetine
10 10
9 9
8 8
Diphenhydramine (ng/mL)
7 7
Fluoxetine (ng/mL)
6 6
5 5
4 4
3 3
2 2
1 1
0 0
0 10 20 30 40 50 0 10 20 30 40 50 60
Gemfibrozil Ibuprofen
12 5
10
4
Gemfibrozil (ng/mL)
Ibuprofen (ng/mL)
8
3
6
2
4
2 1
0 0
0 10 20 30 40 50 0 10 20 30 40 50 60
Time (d) Time (d)
Removal trend of gemfibrozil in aerobic activated Removal trend of ibuprofenin aerobic activated sludge
sludge bioreactor
bioreactor
90
Iopromide Naproxen
10
5.5
5.0
8 4.5
4.0
Iopromide (ng/mL)
Naproxen (ng/mL)
6 3.5
3.0
2.5
4
2.0
1.5
2
1.0
0.5
0 0.0
0 10 20 30 40 50 60 0 10 20 30 40 50 60
Time (d) Time (d)
Removal trend of iopromide in aerobic activated Removal trend of naproxen in aerobic activated sludge
Oxybenzone Primidone
10 12
8 10
Oxybenzone (ng/mL)
Primidone (ng/mL)
8
6
4
4
2
2
0 0
0 10 20 30 40 50 60 0 10 20 30 40 50
Removal trend of oxybenzone in aerobic activated Removal trend of promidone in aerobic activated
Sucralose Sulfamethoxazole
14 10
12
8
Sulfamethoxazole (ng/mL)
10
Sucralose (ng/mL)
8 6
6
4
2
2
0
0
0 10 20 30 40 50 60
0 10 20 30 40 50 60
Time (d) Time (d)
TCEP TCPP
10
12
9
11
8 10
7 9
8
TCEP (ng/mL)
6
TCPP (ng/mL)
7
5
6
4 5
3 4
2 3
2
1
1
0
0
0 10 20 30 40 50 60
0 10 20 30 40 50 60
Time (d)
Time (d)
Removal trend of TCEP in aerobic activated sludge Removal trend of TCPP in aerobic activated sludge
bioreactor bioreactor
92
TDCPP Trimethoprim
10 10
9 9
8 8
Trimethoprim (ng/mL)
7
TDCPP (ng/mL)
6 6
5 5
4 4
3 3
2 2
1 1
0 0
0 10 20 30 40 50 0 10 20 30 40 50 60
Removal trend of TDCPP in aerobic activated sludge Removal trend of trimethoprim in aerobic activated
bioreactor sludge
93
Cin (µg/L)
Cp (µg/L)
50 CRT (d) 30
45
40
OMPs concentration (µg/L)
35
20
30
25
20
10
15
10
0 0
0 5 10 15 20 25 30 35
time (d)
Cin (µg/L)
Cp (µg/L)
CRT (d) 30
50
25
OMPs concentration (µg/L)
40
20
30
15
20
10
10
0
5 10 15 20 25 30 35
time (d)
Cin (µg/L)
Cp (µg/L)
50 CRT (d) 30
45
40
25
20
10
15
10
0 0
0 5 10 15 20 25 30 35
time (d)
Cin (µg/L)
Cp (µg/L)
CRT (d) 60
50
50
OMPs concentration (µg/L)
40
40
30
30
20
20
10 10
0 0
0 5 10 15 20 25 30 35
time (d)
Cin (µg/L)
Cp (µg/L)
CRT (d) 45
60
40
30
40
25
30 20
15
20
10
10
5
0 0
0 5 10 15 20 25 30 35
time (d)
Cin (µg/L)
Cp (µg/L)
40 CRT (d)
30
35
OMOs concentration (µg/L)
30 25
25 20
20
15
15
10
10
5
5
0 0
0 5 10 15 20 25 30 35
time (d)
Cin (µg/L)
Cp (µg/L)
CRT (d)
60
40
30
40
CRT (d)
30 20
20
10
10
0 0
0 5 10 15 20 25 30 35
time (d)