Characterization of organic membrane foulants in

a submerged membrane bioreactor with pre-ozonation

using three-dimensional excitationeemission matrix
uorescence spectroscopy
Ting Liu, Zhong-lin Chen*, Wen-zheng Yu, Shi-jie You
State Key Laboratory of Urban Water Resources and Environments (SKLUWRE), School of Municipal and Environmental Engineering,
Harbin Institute of Technology, No. 73 Huanghe Road, Nangang District, Harbin 150090, PR China
a r t i c l e i n f o
Article history:
Received 19 July 2010
Received in revised form
25 October 2010
Accepted 22 December 2010
Available online 1 January 2011
Keywords:
Organic membrane foulants
Fluorescence spectroscopy
Pre-ozonation
Membrane bioreactor
a b s t r a c t
This study focuses onorganic membranefoulants ina submergedmembranebioreactor (MBR)
process with pre-ozonation compared to an individual MBR using three-dimensional excita-
tioneemission matrix (EEM) uorescence spectroscopy. While the inuent was continuously
ozonated at a normal dosage, preferable organic matter removal was achieved in subsequent
MBR, andtrans-membrane pressure increasedat a muchlower rate thanthat of the individual
MBR. EEM uorescence spectroscopy was employed to characterize the dissolved organic
matter (DOM) samples, extracellular polymeric substance (EPS) samples and membrane fou-
lants. Four main peaks could be identied from the EEM uorescence spectra of the DOM
samples inbothMBRs. Two peaks were associated withthe protein-like uorophores, and the
other ones were related to the humic-like uorophores. The results indicated that pre-ozon-
ation decreased uorescence intensities of all peaks in the EEM spectra of inuent DOM
especially for protein-like substances and caused red shifts of all uorescence peaks to
different extents. The peak intensities of the protein-like substances represented by Peak T
1
and T
2
in EPS spectra were obviously decreased as a result of pre-ozonation. Bothexternal and
internal fouling could be effectively mitigated by the pre-ozonation. The most primary
component of external foulants was humic acid-like substance (Peak C ) in the MBR with pre-
ozonationandprotein-likesubstance(PeakT
1
) intheindividual MBR, respectively. Thecontent
decrease of protein-like substances and structural change of humic-like substances were
observedinexternal foulants fromEEMuorescence spectra due topre-ozonation. However, it
could be seen that ozonation resulted in signicant reduction of intensities but little location
shift of all peaks in EEM uorescence spectra of internal foulants.
2010 Elsevier Ltd. All rights reserved.
1. Introduction
Low-pressure, hollow-ber membrane ltration has been
generally accepted as the most promising technology for
surface water purication in recent years (Huang et al., 2007;
Gray et al., 2008). As the ltration process continues,
a submerged membrane ltration system becomes
a membrane bioreactor (MBR) system because of accumula-
tion of microorganism and organic substances in raw water.
Thus, a submerged MBR system, which combines membrane
* Corresponding author. Tel./fax: 86 451 86283028.
E-mail address: zhonglinchen@163.com (Z.-l. Chen).
Avai l abl e at www. sci encedi r ect . com
j our nal homepage: www. el sevi er . com/ l ocat e/ wat r es
wa t e r r e s e a r c h 4 5 ( 2 0 1 1 ) 2 1 1 1 e2 1 2 1
0043-1354/$ e see front matter 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.watres.2010.12.023
rejection with microorganism biodegradation in a single tank,
was introduced for treatment of drinking water by Li and Chu
(2003). Afterward, much work has been done on the use of
MBR or MBR-coupled technologies for drinking water treat-
ment (Tian et al., 2008; Sagbo et al., 2008).
Recently, an increasing attention has been paid to MBR
process with pre-ozonation used for treating surface water. It
was reported that MBR, especially containing powdered acti-
vated carbon, could be effectively used to remove total alde-
hydes, assimilable organic carbon (AOC) and biodegradable
dissolved organic carbon (BDOC) from pre-ozonated water
(Williams and Pirbazari, 2007; Treguer et al., 2010). Although
the removal of contaminants can be improved by ozone
oxidation, a better understanding of the inuence of pre-
ozonation on membrane fouling process is still needed. The
relative contribution of dissolved organic matter (DOM) to
membrane fouling has been proved in the range of 26e52% in
MBRs (Bouhabila et al., 2001; Lee et al., 2003). Moreover,
extracellular polymeric substances (EPS), which are produced
by bacteria, are reported as one of the factors causing
membrane fouling in MBRs (Chang et al., 2001; Drews et al.,
2006). Therefore, an insight into the impact of pre-ozonation
on DOM and EPS is helpful to develop efcient strategies for
membrane fouling control.
Three-dimensional excitationeemission matrix (EEM)
uorescence spectroscopy has been successfully utilized to
obtain the structural information of organic substances at
a relatively low concentration (Chen et al., 2003;

Swietlik and
Sikorska, 2004b; Gone et al., 2009). It can be used as a simple
and sensitive technique to capture specic uorescence
features which correspond to humic- and protein-like mate-
rials in a single matrix in terms of uorescence intensities
(Hudson et al., 2007). Therefore, EEM uorescence spectros-
copy was employed to investigate the componential differ-
ences of DOM and EPS between the anoxic and oxic phases of
an MBR process (Wang et al., 2009). EEM uorescence spec-
troscopy was also applied to monitor the performance of pre-
treatment stages of membrane systems (Peiris et al., 2010b)
and identify the oxidation-induced structural changes of DOM
fractions of a ltered river water (Zhang et al., 2008).
The great potential of EEM uorescence spectroscopy is
noticed for analysis of membrane foulants. Wang et al. (2009)
found that the dominant uorescence substances in gel layer
(mainly caused by soluble microbial byproduct, colloids,
solutes, etc.) on membrane surface of MBR were protein-like
substances that might be due to the retention of proteins by
the ne pores of the membrane. In addition, Kimura et al.
(2009) demonstrated that EEM uorescence spectra could be
an effective analytical tool for the investigation of physically
irreversible foulants in MBRs under different solid retention
times. Peiris et al. (2010a) combined principal component
analysis and uorescence EEM measurements to characterize
three membrane foulant fractions in the loosely attached
foulants and chemically extracted foulants during UF of
natural river water. According to a reviewby Meng et al. (2010),
membrane fouling mainly results from the accumulation of
retained substances on the membrane surface (i.e., external
fouling) and the deposition of substances in membrane pores
(i.e., internal fouling). Some studies reported that external
fouling or foulant layer formation is the major cause of
membrane fouling in MBRs (Lee et al., 2001; Meng et al., 2007).
On the other hand, the internal fouling or pore-blocking can
lead to the formation of irreversible fouling, which is harmful
for the long-term operation of MBRs (Meng et al., 2010). Thus,
if external and internal foulants are both taken into consid-
eration and analyzed using EEM uorescence spectroscopy, it
will be of great signicance to give an insight into the fouling
behavior in membrane-based water treatment processes.
The aim of the present work is to characterize the organic
membrane foulants in a submerged MBR with pre-ozonation
compared to an individual MBR by EEM uorescence spec-
troscope. The DOMand EPS samples, which are closely related
with membrane fouling, were also analyzed by the EEM uo-
rescence technology. The external and internal foulants in
both MBRs were identied and the comparison between them
was conducted to contribute to a better understanding of
membrane fouling in MBR processes.
2. Materials and methods
2.1. Experimental set-up
An individual MBR process without pre-ozonation (denoted as
MBR-A) and an identical MBR process with pre-ozonation
(denoted as MBR-B) were operated in parallel in this study. A
schematic illustration of MBR-B is shown in Fig. 1. The hollow-
ber UF membrane modules (Litree, China) were made of
polyvinyl chloride (PVC) with a nominal pore size of 0.01 mm
and a total surface area of 0.1 m
2
. The raw water was fed into
a constant-level tank to manipulate the water head for the
subsequent units. Ozone gas generated from an ozone gener-
ator (DHX-1, Jiujiu, China) was continuously bubbled into the
water througha porous glass plate inanozone contact column.
A gas-phase ozone monitor was connected to a side stream
from the generator to measure the ozone concentration. Pre-
ozonated feedwater was then supplied to MBR-B from a reten-
tioncolumnfor further reactionof residual ozone to prevent its
impact on microorganism in MBR (Li et al., 2006). The efuent
was collected directly fromthe membrane module by a suction
pump, and a manometer was xed between the membrane
module and the suction pump to monitor the trans-membrane
pressure (TMP). To supply the oxygen for microbial respiration
and turbulence for membrane surface cleaning, continuous
aeration was provided at the bottom of bioreactor. The exper-
imental set-up of MBR-Awas the same as that of MBR-B except
for the absence of ozonation unit.
2.2. Simulated raw water supply
The raw water was prepared in a way similar to that used by
Tian et al. (2008). Domestic sewage was added to the local tap
water (Harbin, China) of a volumetric ratio of 1:30 to simulate
the surface water supply slightly polluted by sewage
discharge. 1 mg/L of humic acid (Jufeng, Shanghai, China) was
also added to the raw water. The synthesized raw water was
then stabilized for 24 h at room temperature before use.
During the experiment, the raw water was kept at a tempera-
ture in the range of 15.5e18.3

C and the pH in the range of
7.1e7.4; other water quality parameters are listed in Table 1.
wa t e r r e s e a r c h 4 5 ( 2 0 1 1 ) 2 1 1 1 e2 1 2 1 2112
2.3. Operating conditions
MBRs with each effective volume of 1 L were conducted in
a dead-end ltration mode at constant ux. Membrane ux
was predetermined at a relatively low value of 10 L/(m
2
h),
which corresponded to a hydraulic retention time (HRT) of
1.0 h. A30-minoperational cycle for suctionfollowed by 1-min
backwashing with the efuent was controlled by a timer. The
ratio of air to inuent was kept at 20:1 (V:V), and sludge
retention time (SRT) was maintained at 40 d. The two MBRs
fed with rawwater had beenoperated stably for three months,
so that the biomass was accumulated in the reactors for
acclimation to the water. Then the mixed liquid of the two
bioreactors was mixed and shared between them to have the
same initial condition. The MBRs were operated continuously
again with new membranes and pre-ozonation process was
applied to MBR-B at ozone dosage of 1.5 mg/L-raw water. The
HRT of the ozone contact column and the retention column
was 15 min and 20 min respectively. The experiments were
carried out under normal operating conditions (e.g., ozone
dosage and reaction time) commonly adopted in water plants.
2.4. Extraction of EPS from the mixed liquid
EPS were extracted from the mixed liquid in the MBR
according to the thermal treatment method (Chang and Lee,
1998). The mixed liquid was centrifuged for 30 min at
3200 rpm to remove the bulk solution. After the supernatant
was discarded, the remaining pellet was washed and resus-
pended with saline water (0.9% NaCl solution). The mixed
liquid was then subjected to heat treatment (100

C, 1 h) and
centrifuged again under the same operating conditions. The
centrifuged supernatant was EPS solution, which was ltered
through a 0.45 mm acetate ber membrane and used for EEM
uorescence analysis.
2.5. Extraction of foulants
At the end of operation, the fouled membrane modules were
taken out from the reactors when the TMP exceeded 35 kPa.
The external foulants on membrane surface were carefully
scraped off with a plastic sheet (Deli, China) and simulta-
neously ushed with deionized (DI) water. The collected
water line
air line
ozone gas line
Fig. 1 e Schematic diagram of MBR-B (1-feed pump; 2-high level tank; 3-constant level tank; 4-ozone contact column;
5-retention column; 6-bioreactor; 7-membrane module; 8-manometer; 9-efuent pump; 10-backwash pump; 11-ozone
generator; 12-gas phase ozone analyzer; 13-ozone gas owmeter; 14-ozone destruction unit; 15-air blower; 16-air
owmeter; 17-air diffuser; 18-sludge discharge valve).
Table 1 e Pollutants removal efciencies of MBR-A and MBR-B.
Water quality
indexs
Raw water After
pre-ozonation
MBR-A MBR-B
Efuent Total
removal (%)
Efuent Total
removal(%)
Turbidity (NTU) 2.31 1.00 2.09 0.60 0.07 0.01 97.0 1.3 0.07 0.01 97.0 1.2
COD
Mn
(mg/L) 4.25 0.27 3.15 0.22 2.30 0.28 45.9 3.7 1.60 0.20 62.4 3.3
DOC (mg/L) 6.020 0.784 5.497 0.572 4.336 0.603 28.0 6.3 3.542 0.568 41.2 6.2
UV
254
(cm
1
) 0.077 0.003 0.046 0.002 0.059 0.002 23.4 2.8 0.036 0.002 53.2 3.0
wa t e r r e s e a r c h 4 5 ( 2 0 1 1 ) 2 1 1 1 e2 1 2 1 2113
sample was fully mixed using a magnetic blender (JB-2, Leici,
China) at 200 rpmfor 1 h, it was thenltered througha 0.45 mm
acetate ber membrane for EEM uorescence analysis (Wang
et al., 2009). After the membrane surface was wiped with
a sponge, 0.01 mol/L NaOH was used for extraction of internal
foulants and the bers were soaked for 24 h at 20

C in the
alkaline solution according to the method described by
Kimura et al. (2009).
2.6. Analytical methods
2.6.1. Water quality analysis
Water quality analysis was conducted following the standard
methods (APHA, 1998). Turbidity was monitored by a turbi-
dimeter (Turbo550, WTW, Germany). The dissolved organic
carbon (DOC) was measured by a total organic carbon (TOC)
analyzer (TOC-VCPH, Shimadzu, Japan). COD
Mn
was analyzed
by the potassium permanganate oxidation methods. UV
absorbance at 254 nm (UV
254
) was determined by using
a spectrometer (T6, Puxi, China).
2.6.2. Three-dimensional excitationeemission matrix (EEM)
uorescence spectroscopy
Fluorescence measurements were conducted using a spectro-
uorometer (FP-6500, Jasco, Japan) equipped with a 150 W
xenon lamp at ambient temperature of 24

C. A 1-cm quartz
cuvette with four optical windows was used for the analyses.
Emission scans were performed from 220 to 550 nm at 5 nm
steps, with excitation wavelengths from220 to 450 nmat 5 nm
intervals. The detector was set to high sensitivity, and the
scanning speed was maintained at 2000 nm/min in this study;
the slit widths for excitation and emission were 5 nm and
3 nm respectively. Under the same conditions, uorescence
spectra for Milli-Q water were subtracted from all the spectra
to eliminate water Raman scattering and to reduce other
background noise. During the course of uorescence analysis,
the Raman scattering peak intensity for Milli-Q water (exci-
tation at 350 nm, emission at 400 nm) was recorded as
a standard to verify the instrument stability. Mean intensity of
the Raman peak was 36.10 units and the differences were less
than 2%, conrming that there were no signicant uctua-
tions in the performance of the spectrouorometer
throughout the experimental period. The EEM spectra are
plotted as the elliptical shape of contours. The X-axis repre-
sents the emission spectra while the Y-axis represents the
excitation wavelength, and the third dimension, i.e., the
contour line, is given to express the uorescence intensity.
3. Results and discussion
3.1. Process performance
As shown in Table 1, the turbidity was reduced from
2.31 1.00 NTUto a level as low as 0.07 0.01 NTUfor the two
MBRs, which indicated an excellent performance of particle
removal. The organic matter removal efciencies in terms of
COD
Mn
, DOC and UV
254
in both MBR-A and MBR-B processes
are summarized in Table 1. The results showed that the
remarkably improved performance in organic matter removal
was ascribed to pre-ozonation implementation in MBR-B.
Especially, an approximately 30% higher UV
254
decrease was
achieved in MBR-B (53.2 3.0%) compared to that in MBR-A.
Three functions were identied for the contributions to
removal of organic contaminants in MBR-B: partial degrada-
tion or complete mineralization by ozone oxidation, physical
retention by UF membrane, and biodegradation by active
biomass within the reactor. It should be noted that the organic
matter removal was attributed to the synergetic effect of these
three functions.
It can be seen from Fig. 2 that TMP increased with opera-
tion time while the membrane ux was maintained constant
at about 10 L/(m
2
h) during the experiment before TMP
exceeded 30 kpa. As a two-step fouling phenomenon, the TMP
variations exhibited a slow increase followed by a rapid
increase. The TMP gradually increased with time from the
initial 5 kPa for both systems with a similar trend within the
beginning phase (0e8 days). However, there was a distinct gap
of 1.5 kPa between them on Day 16. The permeate ux obvi-
ously declined when the TMP of MBR-A reached 35 kPa, and
this operation process came to an end for a further analysis of
membrane foulants. The nal TMP of MBR-B only increased
to 23.5 kPa, which was 12.5 kPa lower than that of MBR-A
(36 kPa). It was thus believed that ozone pre-oxidation was an
effective pre-treatment strategy to reduce the increasing rate
of TMP to lower energy requirement for the membrane
ltration process at a constant ux. In order to identify the
proportions of external and internal fouling resistances, the
membranes were taken out from the reactors at the end of
experiment and the external foulants were removed from
membrane surface. The membrane modules were reinstalled
into the bioreactors and internal fouling resistances were
evaluated. The TMP in MBR-Aand MBR-B were then decreased
to 12 kPa and 9 kPa respectively. Compared to those of 77.4%
and 22.6%in MBR-A, the contributions of external and internal
fouling resistances to TMP development were 78.4%and 21.6%
in MBR-B. Therefore, it can be concluded that the proportions
of external and internal fouling resistances were similar in
both MBRs, suggesting that ozone pre-oxidation was able to
alleviate both of the two kinds of membrane fouling.
0 10 20 30 40 50
0
5
10
15
20
25
30
35
40
45
50
Time (day)
T
M
P

(
k
p
a
)
0
2
4
6
8
10
12
TMP (MBR-A)
TMP (MBR-B)
Flux (MBR-A)
Flux (MBR-B)
F
l
u
x

(
L
/
(
m
2
h
)
)
Fig. 2 e Comparison of TMP and membrane ux variations
in MBR-A and MBR-B.
wa t e r r e s e a r c h 4 5 ( 2 0 1 1 ) 2 1 1 1 e2 1 2 1 2114
3.2. EEM uorescence spectra of DOM in the two systems
Three-dimensional EEM uorescence spectroscopy has been
successfully utilized to identify the chemical composition of
DOM because of its ability to distinguish among certain
classes of organic matter in natural waters (Saadi et al., 2006).
In UF application, the major membrane foulant is natural
organic matter, which contains a complex mixture of humic
and fulvic acids, proteins, carbohydrates of various molecular
size and functional groups (Her et al., 2003; Saravia et al.,
2006). There are ve key uorescence peaks referred to as
uorophores A, C, T
1
, T
2
and B commonly observed in fresh-
water samples (Coble, 1996; Baker, 2001). Peak A and C are
related to humic-like substance derived from the breakdown
of plant material (Lee et al., 2008); protein-like uorophores
including tryptophan-like (Peak T ) and tyrosine-like (Peak B)
materials, are usually detected at enhanced levels in water
impacted by domestic sewage (Baker et al., 2003). As shown in
Fig. 3, Peak B has relatively lower uorescence intensity for the
DOM samples, so the other four peaks including Peak A, C, T
1
and T
2
which are distinctly identied were investigated in this
section. The uorescence parameters including peak loca-
tions, uorescence intensity, and different peak intensity
ratios were extracted from EEM uorescence spectra and
summarized in Table 2, which could be employed for quan-
titative analysis.
Generally, intensity reduction of the uorescence peak
between raw water and treated water is an indication for
degradation of uorescing material. It canbe seen fromTable 2
that ozonation approximately decreased the uorescence
intensities of Peak Aand C by 30e40%and those of Peak T
1
and
T
2
by 60e70% for DOM in raw water. Consequently, the uo-
rescence intensities of Peak A and C in EEM spectra of MBR-B
efuent were nearly the same percentage lower than those of
MBR-Aefuent. Meanwhile, the intensities of Peak T
1
and T
2
in
EEM spectra of MBR-B efuent were 40% and 53% lower than
those of MBR-A efuent. The peak intensity ratios are shown
as ratios to Peak C, as this component is considered to be
present in a wide range of water environments (Henderson
et al., 2009). Presented in this way, the data reect the differ-
ences in composition rather than the considerable differences
in concentration. Peak T
1
and C in EEM spectra of DOM
samples, which indicate protein- and humic acid-like
substance respectively, can be referred to as biodegradable
and nonbiodegradable DOM (Reynolds, 2002; Wang et al.,
2009). Since the feedwater synthesized to simulate the
Fig. 3 e EEM uorescence spectra of (a) the inuent (raw water) and (b) the efuent DOM of MBR-A, (c) the inuent (after pre-
ozonation) and (d) the efuent DOM of MBR-B.
wa t e r r e s e a r c h 4 5 ( 2 0 1 1 ) 2 1 1 1 e2 1 2 1 2115
surface water polluted by sewage discharge had a relatively
higher biodegradability, some biodegradable DOM was
mineralized during the ozonation process. The intensity ratio
of Peak T
1
/Peak C decreased in both bioreactors as shown in
Table 2, suggesting that the biodegradable DOM with uo-
rescence was gradually metabolized by microorganism. The
intensity ratio of Peak T
1
/Peak C of MBR-B efuent was 0.46
compared to 0.60 of MBR-A efuent. MBR-B process thus dis-
played a greater capacity for biodegradable DOM removal and
this is benecial to biological stability of treated water and
restraint of bacterial regrowth in distribution system. The
differences in peak intensity ratios of EEM uorescence
spectra in the two MBR systems imply that ozone oxidation is
responsible for the compositional variation of the uorescent
compounds in DOM samples.
The location shift of uorescence peak provides spectral
information on the chemical structure changes of DOM
samples. After ozonation, the locations of the four uores-
cence peaks shifted toward longer wavelength (red shift) to
different extents on the emission and/or excitation scale, and
this observation is in line with Chen et al. (2002). As reported
by

Swietlik et al. (2004a), the ozonation of hydrophobic acid
(HOA) and hydrophilic neutral (HIN) produces carboxylic
acids, aldehydes and ketones, and this may cause the
formation of oxidation byproducts with double bond-con-
taining substituents. A red shift is related to the increase of
carbonyl, hydroxyl, alkoxyl, amino, and carboxyl groups in the
structures of uorophores (Chen et al., 2002; Uyguner and
Bekbolet, 2005), while a blue shift is ascribed to the elimina-
tionof particular functional groups such as carbonyl, hydroxyl
and amine, a reduction in the degree of p-electron systems,
and the decrease in the number of aromatic rings and conju-
gated bonds in a chain structure (

Swietlik et al., 2004a). In the

two systems, the locations of Peak A and T
2
in EEM spectra of
efuent DOM were all red-shifted (5e15 nm) to longer wave-
lengths than those of inuent DOM, while Peak T
1
of efuent
DOM showed a blue shift (5e10 nm) on the excitation scale
compared to that of inuent DOM. The location of Peak C
showed different shift trends in the two systems with respect
to the emission axis. Wang et al. (2009) observed that Peak T
1
and T
2
respectively demonstrated a blue and red shift of the
efuent DOMcompared to those of the inuent DOMin a MBR
for wastewater treatment, which is in good agreement with
the results of both MBRs in this study. Furthermore, they
described that the location of Peak C of the efuent DOM was
red-shifted along the excitation axis and blue-shifted along
the emission axis. This nding agrees with the experimental
results of MBR-A but disagrees with those of MBR-B obtained
during this study. It was therefore assumed that this
difference between the two MBRs was a consequence of
structural changes of the humic-like substances in the raw
water during ozonation process. As a matter of fact, it is likely
that some of the EEM peak shifts result from the changes in
concentration of one of the several overlapping components
(Stedmon et al., 2003; Peiris et al., 2010a). For the purpose of
identifying structural changes, the EEM uorescence intensi-
ties were normalized with respect to the highest peak inten-
sity. Fig. 4 shows the normalized intensities of EEM
uorescence spectra of DOM in raw water and pre-ozonated
water on the excitation scale (emission at 420 nm). When the
intensity of Peak C reduced signicantly due to the ozonation
process, the visual location of Peak A would have a blue shift
to a lesser degree, which was dependent on concentration
changes. Nevertheless, Peak A showed a red shift of 10 nm on
the contrary, which means that the peak shift was necessarily
caused by the structural changes of this kind of humic-like
substances (Peak A). The peak locations in EEM spectra of the
efuent DOMof MBR-B also demonstrated some differences in
comparison with those of the efuent DOM of MBR-A.
3.3. EEM uorescence spectra of EPS in mixed liquid of
the two MBRs
As shown in Fig. 5, the intensity of Peak B in the EEMspectra of
EPS was signicantly enhanced in both MBRs, suggesting that
the organic substances indicated by Peak B were closely
related with metabolic activity of microorganism. Anewpeak,
Table 2 e Fluorescence spectral identications of DOM samples in MBR-A and MBR-B.
Process Samples Peak A Peak C Peak T
1
Peak T
2
Peak Int. ratio
Ex/Em Int. Ex/Em Int. Ex/Em Int. Ex/Em Int. A/C T
1
/C C/T
2
MBR-A Inuent (Raw) 240/405 311.7 310/420 298.3 280/340 283.2 225/335 535.7 1.04 0.95 0.56
Efuent 245/420 304.5 315/415 199.0 270/340 119.5 230/345 206.9 1.53 0.60 0.96
MBR-B Inuent (Ozonated) 250/420 230.5 325/415 170.7 280/345 115.0 230/340 154.2 1.35 0.67 1.11
Efuent 255/425 213.7 330/425 154.3 275/345 71.2 230/345 96.5 1.38 0.46 1.60
Int.: intensity.
Fig. 4 e Normalized intensities of EEM uorescence spectra
of DOM in raw water and pre-ozonated water on the
excitation scale (emission at 420 nm).
wa t e r r e s e a r c h 4 5 ( 2 0 1 1 ) 2 1 1 1 e2 1 2 1 2116
i.e., Peak D, was present at the excitation/emission wave-
lengths (Ex/Em) of 270e280/300e310 nmin EEMspectra of EPS.
According to the ve regions of EEM divided by Chen et al.
(2003), Peak D indicates soluble microbial byproduct (SMP)-
like substances (Region IV). It could be seen that protein- and
SMP-like substances are dominant among uorescent organic
matters in EPS from both MBRs. Compared with the EEM
spectra of DOM samples (Fig. 3), intensity of Peak T
2
in EEM
spectra of EPS was much weaker than that of raw water,
whichmeans that the uorescent DOMrepresented by Peak T
2
in efuents were originated from raw water rather than EPS.
The locations of Peak A and C in EEM spectra of EPS were both
red-shifted by 15e35 nm along the two axes compared to
those in Table 2, indicating that the structure and components
of humic-like substances in EPS were different from those in
DOM samples. Sheng and Yu (2006) found three uorescence
peaks including Peak T
1
, T
2
and C were present in EPS spectra
from a conventional activated sludge system. Wang et al.
(2010) identied Peak T
1
, C, together with a new peak associ-
ated with humic acid-like substances at the Ex/Emof 415e420/
470e475 nm in EPS spectra of an MBR. In this study, three
main peaks and three lesser peaks were observed in EPS
spectra. The organic matters indicated by Peak T
1
and C are
extensively present in EPS samples extracted from various
origins. The locations of the two peaks in EPS spectra in this
study, which located at the Ex/Em of 280/350 nm and
340/435 nm, were similar to those reported by Sheng and Yu
(2006) but different from those observed by Wang et al.
(2009). The differences might be attributed to the fact that
the EPS samples were extracted from different origins and
thus the components in EPS were chemically different.
The intensity of Peak B did not show any decline in EPS
spectra of MBR-B (Fig. 5b) due to pre-ozonation compared to
that of MBR-A (Fig. 5a). It may indicate that the protein-like
substances represented by Peak B were excreted by microor-
ganism in MBRs, and pre-ozonation had little effect on the
metabolic level related to this compound. The peak intensities
of protein-like substances represented by Peak T
1
and T
2
in
EPS spectra of MBR-B were decreased signicantly resulted
from pre-ozonation. Cho et al. (2005) established a functional
equation in which the specic cake resistance was propor-
tional to the EPS concentration. Ahmed et al. (2007) also
observed that as EPS concentration rose, the specic cake
resistance increased, and this consequently resulted in the
rise of TMP. These investigations showed that there is a close
relationship between EPS and the resistance of cake layer on
membrane surface. It can therefore be concluded that the
protein-like substances represented by Peak T
1
and T
2
in EPS
might contribute more to external fouling.
3.4. EEM uorescence spectra of membrane foulants
The amount and composition of organic membrane foulants
were related to the interaction betweenorganic substance and
membrane. The UF membrane used in this study is made of
Polyvinyl chloride (PVC), which is a hydrophobic material.
According to the manufacturer, doping technology is used to
improve its hydrophilicity for higher ux and some other
physical properties. The contact angle reects the hydro-
phobic/hydrophilic character of membranes. The PVC
membrane has an average contact angle of 68 2

(provided
by the manufacturer). On the one hand, ozone oxidation is
able to increase the polarity and hydrophilicity of the
substances to make hydrophobic membranes less susceptible
to adsorptive fouling. In this study, membrane fouling was
reduced by pre-ozonation, which means that the PVC
membrane might still exhibit hydrophobic property. On the
other hand, ozonation reduced the amount of humic
substances because of their breakdown to lower molecular
weight (MW) compounds. Therefore, although the size of
some DOM molecules before or after pre-ozonation was
smaller than the nominal pore size of PVC membrane
(0.01 mm), biodegradation of these low-MW ozonation prod-
ucts caused less accumulation of foulants on membrane
surface and/or in membrane pores.
Unlike conventional methods such as the ratio of carbohy-
drate toprotein(C/P) whichis incompetent tofullycharacterize
membrane foulants, EEM uorescence analysis is able to
provide more useful information on the characteristics of
organic membrane foulants (Kimura et al., 2009). Hence, both
external and internal foulants were extracted from the fouled
membranes and analyzed by using EEM uorescence spec-
troscopy at the end of operation.
a
b
Fig. 5 e EEM uorescence spectra of EPS extracted from
(a) MBR-A and (b) MBR-B.
wa t e r r e s e a r c h 4 5 ( 2 0 1 1 ) 2 1 1 1 e2 1 2 1 2117
3.4.1. External foulants
Fig. 6 shows EEM uorescence spectra measured for external
foulants extracted from the fouled membranes of the two
MBRs. Peak T
1
predominated the EEM uorescence spectra of
external foulants in MBR-A, while Peak C was dominant in the
EEMuorescence spectra of external foulants in MBR-B. It was
demonstrated that the external foulants were composed of
the protein-like substances represented by Peak T
1
and the
humic acid-like substances represented by Peak C as the most
primary components in MBR-A and MBR-B, respectively. The
comparison in TMP development of both MBRs showed that
membrane fouling of MBR-A was more serious than that of
MBR-B. As stated in Section 3.1, the contribution of external
fouling resistance to TMP development reached 75e80%in the
two systems. Therefore, it can be concluded that protein-like
substances rather than humic acid-like substances contrib-
uted more to the external fouling resistance. This is consistent
with the ndings of Hong et al. (2007) and Drews et al. (2007),
who reported that proteins could induce severe membrane
fouling as one of the major components in membrane fou-
lants. The appearance of dominant protein-like peak in
external foulants in MBR-A indicated the accumulation of
protein-like substances on the membrane surface, whereas
the intensity of protein-like peak was weakened to a signi-
cant degree in MBR-B which could be attributed to ozonation.
Thus, it was reasonable to infer that the reduced accumula-
tion of protein-like substances into gel layer may play an
important role against TMP increase in MBR-B at constant
ux.
The locations of peak T
1
(Ex/Em 280/345) and peak T
2
(Ex/Em 230/335) in the EEM uorescence spectra of external
foulants from MBR-B were similar to those of external fou-
lants from MBR-A as shown in Fig. 6. However, the location of
Peak A in the EEM uorescence spectra of external foulants of
MBR-A was red-shifted by 10 nm along the excitation axis and
blue-shifted by as much as 25 nm along the emission axis
compared to that of external foulants of MBR-B. The location
of Peak C of MBR-A external foulants was blue-shifted to
shorter wavelengths than that of MBR-B external foulants.
These observations implied that the structures of humic-like
substances represented by Peak A and C in the external fou-
lants of the two MBRs differed from each other.
In conclusion, the content decrease of protein-like
substances and the structural change of humic-like
substances were observed in external foulants from EEM
uorescence spectra due to pre-ozonation. The study carried
out by Schlichter et al. (2003) indicated that continuous addi-
tion of ozone caused a drastic reduction in adsorption-
induced membrane fouling during the UF of humic acid
solution. Nevertheless, Her et al. (2007) reported that nano-
ltration (NF) membrane fouling increased mainly due to the
adhesive EPS released by algae upon ozonation. It may actu-
ally be attributed to the average size of NF membrane pores
which is smaller than that of UF membrane pores. Moreover,
it may also be attributed to the presence of abundant algae in
the raw water used for their study in that season, which
indicated the unsuitability for application of pre-ozonation.
They also found that ozonation showed opposite results for
humic- and protein-like substances as for UVabsorbance ratio
index (UVA
210
/UVA
254
), which provides information on the
relative proportions between UV-absorbing functional groups
and unsaturated compounds in NOM. The results of our study
coincide with the results of their investigations in this respect.
It is possible for ozonation to reduce the number of unsatu-
rated groups and form new groups to cause the structural
changes of humic substances with large molecular size.
Ozonation may also destroy the particular functional groups
of protein-like substances to thereby result in a relatively
lower level of their characteristics.
3.4.2. Internal foulants
Membrane bers of the two MBRs with the same quantity
were chemically treated to extract the internal pore foulants
for investigation. In order to identify the difference of DOM
characteristics of internal foulants caused by ozone pre-
oxidation, analysis of EEM uorescence spectra of internal
foulants was carried out and the results are shown in Fig. 7.
Peak C at Ex/Em 440e445/275 nmpredominated in both EEM
uorescence spectra, indicating that the humic acid-like
substances represented by Peak C were the dominant
components of the internal foulants. The characteristics of
EEM uorescence spectra of internal foulants obviously
differed from those of external foulants. The uorescence
intensities of the four peaks of internal foulants from MBR-B
were much weaker than those of internal foulants from
a
b
Fig. 6 e EEM uorescence spectra of external foulants
extracted from (a) MBR-A and (b) MBR-B.
wa t e r r e s e a r c h 4 5 ( 2 0 1 1 ) 2 1 1 1 e2 1 2 1 2118
MBR-A, which was in line with the tendency of internal
fouling resistances indicated by TMP increase. By comparison
of the EEM uorescence spectra of internal foulants from the
two MBRs, it could be seen that the four main peaks were
similarly located with a difference of no more than 5 nmalong
the two axes.
The results indicated that the pre-ozonation process had
the potential to effectively mineralize some NOM with small
molecular size in raw water. For the NOM with relatively
higher molecular size, pre-ozonation could cleave unsatu-
rated bonds in aromatic moieties and reduce the molecular
size/weight of the substances to make them more amenable
to microbial degradation and utilization. The less quantity of
organic matter tended to deposit or adsorb into membrane
pores which induced internal fouling, suggesting that ozona-
tion conducted well as a pre-treatment process for the
biodegradation by microorganism in MBR-B. It was likely that
the structures of organic substances in internal foulants
changed slightly, and almost the same proportion of the
organic matter content decreased as a result of ozone
oxidation.
4. Conclusions
Two identical submerged MBRs with or without pre-ozonation
were comparatively operated to investigate performance of
the processes and characterize organic membrane foulants
using EEM uorescence spectroscopy. It can be seen that
preferable organic matter removal was achieved in the MBR
process with pre-ozonation, and its TMP increased at a rate
much lower than that of the individual MBR. EEMuorescence
spectroscopy was employed to characterize the DOMsamples,
EPS samples and membrane foulants of both MBRs. The
results indicated that pre-ozonation decreased uorescence
intensities of the four main peaks in the inuent DOM spectra
especially for protein-like substances and caused red shifts of
all uorescence peaks to different extents. The peak intensi-
ties of the protein-like substances represented by Peak T
1
and
T
2
in EPS spectra were obviously decreased as a result of pre-
ozonation. Both external and internal fouling could be effec-
tively mitigated by the pre-ozonation. The most primary
components of external foulants were humic acid-like
substance (Peak C ) in the MBR with pre-ozonation and
protein-like substance (Peak T
1
) in the individual MBR,
respectively. The content decrease of protein-like substances
and structural change of humic-like substances were
observed in external foulants from EEM uorescence spectra
due to pre-ozonation. However, it could be seen that ozona-
tion resulted in signicant reduction of intensities but little
change of locations of all peaks in EEMuorescence spectra of
internal foulants. Further work is required to assess the
impact of pre-ozonation on other kinds of DOM, such as
polysaccharide substances, to extend the knowledge of
fouling control of MBR processes.
Acknowledgements
This research was funded by National High Technology
Research and Development Program of China (2007AA06Z339)
and State Key Laboratory of Urban Water Resource and Envi-
ronment (HIT, Grant No. 2010DX12).
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