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PAPER
Westerhoff et al.
Occurrence and removal of titanium at full scale wastewater treatment plants:
implications for TiO2 nanomaterials.
 Journal of Dynamic Article Links < C
Environmental
Monitoring
Cite this: J. Environ. Monit., 2011, 13, 1195
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Occurrence and removal of titanium at full scale wastewater treatment plants:

implications for TiO2 nanomaterials†‡
Paul Westerhoff,*a Guixue Song,xa Kiril Hristovskib and Mehlika A. Kiser{a
Received 10th January 2011, Accepted 22nd March 2011
DOI: 10.1039/c1em10017c

Titanium dioxide nanoparticles increasingly will be used in commercial products and have a high
likelihood of entering municipal sewage that flows to centralized wastewater treatment plants (WWTPs).
Treated water (effluent) from WWTPs flows into rivers and lakes where nanoparticles may pose an
ecological risk. To provide exposure data for risk assessment, titanium concentrations in raw sewage and
treated effluent were determined for 10 representative WWTPs that use a range of unit processes. Raw
sewage titanium concentrations ranged from 181 to 1233 mg L1 (median of 26 samples was 321 mg L1).
The WWTPs removed more than 96% of the influent titanium, and all WWTPs had effluent titanium
concentrations of less than 25 mg L1. To characterize the morphology and presence of titanium oxide
nanoparticles in the effluent, colloidal materials were isolated via rota-evaporation, dialysis and
lyophilization. High resolution transmission electron microscopy and energy dispersive X-ray analysis
indicated the presence of spherical titanium oxide nanoparticles (crystalline and amorphous) on the
order of 4 to 30 nm in diameter in WWTP effluents. This research provides clear evidence that some
nanoscale particles will pass through WWTPs and enter aquatic systems and offers a methodological
framework for collecting and analyzing titanium-based nanomaterials in complex wastewater matrices.

a
School of Sustainable Engineering and The Built Environment, Civil,
Environmental and Sustainable Engineering Program, Ira A. Fulton
Schools of Engineering, Arizona State University, Engineering Center,
G-Wing Room ECG252, Box 5306, Tempe, AZ, 85287-5306, USA.
E-mail: p.westerhoff@asu.edu; Fax: +1 480-965-0557; Tel: +1 480-965-
2885
b
Environmental Technology, Department of Applied Sciences and
Mathematics, College of Technology and Innovation, Arizona State
University—Polytechnic Campus, 6073 S. Backus Mall, Mesa, AZ,
85212, USA. E-mail: Kiril.Hristovski@asu.edu; Tel: +1 480 727 1291
† Published as part of a JEM themed issue dedicated to significant recent
advances in Environmental Nanotechnology: Guest Editor Professor
Wunmi A. Sadik.
‡ Electronic supplementary information (ESI) available: See DOI:
10.1039/c1em10017c
x Tel: +1 480 284 9348; Fax: +1 480 965 0557; E-mail: gxsong@gmail.
com
{ Tel: +1 480 965 3589; Fax: +1 480 965 0557; E-mail: ayla@asu.edu
Introduction
Engineered nanomaterials may pose an ecological risk to aquatic
organisms.1–9 Risk assessment and management requires infor-
mation on both toxicity and exposure. Hundreds of studies now
exist on the potential toxicity of nanomaterials, but only a few
exposure modeling papers and even fewer actual exposure
sampling studies have been reported. This paper aims to expand
our knowledge regarding release of nanomaterials, specifically
titanium-based nanoparticles, from municipal wastewater treat-
ment plants (WWTPs) into receiving waters (lakes, rivers,
streams). WWTPs have been identified as a major point source
for engineered nanomaterials entering into aquatic systems.10–15
Research to determine the potential removal mechanisms for
engineered nanomaterials during wastewater treatment has only

Environmental impact
Engineered nanomaterials may pose an ecology risk to aquatic organisms. Risk assessment and management requires information
on both toxicity and exposure. Many studies exist regarding toxicity of nanomaterials, but only a few exposure assessments have
been reported. Municipal wastewater treatment plants (WWTPs) are a major point source for engineered nanomaterials entering
into aquatic systems. This paper aims to expand our knowledge regarding discharge of nanomaterials, specifically titanium-based
nanoparticles, from WWTPs into lakes, rivers and streams. The conclusions include data on removal efficiency of different WWTP
unit processes, quantification of titanium in WWTP effluents, microscopy validation that titanium dioxide nanoparticles are present
in WWTP effluents and analytical methods to recover and characterize nanomaterials from complex water matrices such as WWTP
effluents.

This journal is ª The Royal Society of Chemistry 2011 J. Environ. Monit., 2011, 13, 1195–1203 | 1195
 recently begun.16–25 These studies suggest that among different
types of nanomaterials, titanium dioxide (TiO2) should occur at
the highest concentration, and therefore TiO2 is the focus herein.
Titanium (Ti) is the ninth most abundant element and the
seventh most abundant metal in the Earth’s crust.26 With
significant worldwide reserves in excess of 600 million tons, the
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estimated annual production of Ti metal is 90 000 tons, and the
annual production of titanium dioxide (TiO2) is approximately
4.3 million tons.27 Although Ti has numerous industrial appli-
cations, from metal alloys to aerospace technologies to
biomedical devices, approximately 95% of mined Ti is refined
into nearly pure TiO2 through the treatment of Ti-bearing ores
with carbon, chlorine, oxygen or sulfuric acid.28 Titanium
dioxide is widely used in industry, commercial, residential and
personal care products from paint to paper to toothpaste.29
Much of the TiO2 used today is in the form of nanoparticle
aggregates or micron-sized materials (i.e., bulk TiO2 products),
but industry trends suggest much higher usage of nano-TiO2 in
the near future.30 Titanium dioxide is one of the most common
nanomaterials used in nano-based products.31 Recent analyses
suggest that nanoscale TiO2 will begin to dominate the bulk TiO2
market because the nanoscale materials are transparent to visible
light, are highly UV absorbent, have an iridescent quality, and
are photocatalyst.30,32 The surface properties of TiO2 are often
modified to improve its dispersion in products or to suppress
undesirable toxic effects,32 and it has been shown that surface
properties affect the fate and toxicity of nanomaterials in
WWTPs and aquatic systems.33–40 Despite a wide number of
analytical techniques capable of size separating nanomaterials
(e.g., field flow fractionation, hydrodynamic separation, centri-
fugation, ultrafiltration) prior to quantification, few have actu-
ally been applied to full-scale WWTP samples. Likewise, the
ability to isolate titanium oxide nanomaterials from WWTP
effluent samples for electron microscopy characterization
remains a significant technical challenge.
Previously, we monitored for titanium over time and between
unit processes at one WWTP. To assess the representative nature
of that facility, we now report titanium concentrations at the
same facility along with nine other WWTPs representing a broad
spectrum of different types of unit processes. Raw sewage and
Fig. 1 General wastewater treatment process flow diagram indicating removal mechanisms for titanium nanoparticles.
1196 | J. Environ. Monit., 2011, 13, 1195–1203
effluent samples were collected and analyzed for titanium.
Because of low titanium concentrations in the effluents and
a desire to obtain solid-state (lyophilized) samples from the
effluents for electron microscopy characterization, large volume
samples (20 to 30 litres) were collected, concentrated via rota-
evaporation, and purified via dialysis prior to freeze drying
(lyophilization). Titanium concentrations in the solids after
lyophilization were quantified by ICP, and solids were analyzed
by transition electron microscopy (TEM) with energy dispersive
X-ray (EDX) analysis to confirm the presence of titanium oxide
(TiOx) nanoparticles. We also discuss removal mechanisms for
nanoscale titanium dioxide across the different types of unit
processes at the 10 WWTPs sampled.
Methods
Description of field sites and sampling protocol
Samples were collected between April and August 2009 from 10
full-scale municipal WWTPs from southern to central Arizona.
The facilities ranged in size, with flows from 0.1 to 7 m3 s1. The
WWTPs employed a range of different biological treatment
processes (Fig. 1 and Table 1): conventional activated sludge,
trickling filter, microfiltration (0.1 mm), membrane bioreactor
(MBR), nitrification/denitrification, or various combinations.
The water quality of each effluent determined using methods
reported elsewhere41 is summarized in the ESI (Table S1‡). Grab
samples were collected from the headworks and effluent (prior to
disinfection) at each WWTP. Samples for titanium detection
were collected in acid washed 250 mL Nalgene bottles. Samples
for colloidal separation were collected in clean 20 litre carboys.
Colloid isolation
Two types of colloids were isolated using rota-evaporation fol-
lowed by dialysis using established methodologies:41–43 (1) inor-
ganic plus organic colloids (HCl-colloids) isolated by dialysis
against hydrochloric acid (HCl), which removed carbonate and
acid-soluble salts,44 and (2) HCl–HF-colloids isolated by dialysis
of HCl-colloids with hydrofluoric acid (HF) to remove silica and
other inorganics. Effluent samples (20 to 30 L) were filtered
This journal is ª The Royal Society of Chemistry 2011
 Table 1 Occurrence of titanium in WWTP headworks and treated effluent

Titanium content of water/mg Ti L1

Treated effluent
Effluent (based upon
Facility Primary gravity Secondary gravity Other separation COD/mg L1 Treated HCl-colloid
ID sedimentation Biological treatment sedimentation processes (TDN) Headworksa effluenta separation)b
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1 3 Activated sludge 3 — 19 (3 mg N L1) 615  538 53 3.8

Microfiltration <2
Reverse osmosis <2
2 3 Activated sludge 3 Tertiary filtration 9 (5.3 mg N L1) 180  30 77 0.6
3 3 Activated sludge 3 19 (4.8 mg N L1) 363  119 31 1.1
4 3 Activated sludge 3 16 (5.6 mg N L1) 141  28 22 0.4
5 3 Activated sludge 3 18 (15 mg N L1) 581  125 18  3 2
and lagoons
6 3 Pure oxygen 3 40 (37 mg N L1) NA 84 5.4
activated sludge
7 3 Activated sludge 3 34 (3.4 mg N L1) 233  57 21 6.2
8 3 Trickling filter 3 29 (34 mg N L1) 549  57 13  3 7.1
9 3 Activated sludgec Submerged 18 (5.2 mg N L1) 310  23 <2 0.2
microfiltration
membrane
10 3 Activated sludgec Submerged 21 (4.8 mg N L1) 422  40 42 0.8
microfiltration
membrane
a
Analyzed for titanium after digestion of water samples (MDL of 0.5 mg Ti L1). b Analyzed for titanium after digestion of lyophilized HCl-colloid
separation samples (detection limit <0.1 mg L1 because large mass could be used). c Total suspended solids concentration of activated sludge in
membrane bioreactor systems is 3 to 5 times larger than activated sludge in WWTPs with secondary gravity separation.

immediately after delivery to the laboratory through a Balston
inline glass-fiber filtration system (100-25-AH, nominal pore size
0.9 mm). The filtrate was kept at 4  C before rota-evaporation. The
sample was adjusted with 6 M HCl to pH 4 and concentrated using
a Buchi Rotavapor 220 to a salt slurry (200 to 500 mL). The slurry
was loaded in a 3500 Da dialysis tube and dialyzed against 4 L of
0.1 M HCl until all salts were visually dissolved and the color of
the permeate solution was negligible; the permeate was then dis-
carded. The HCl-treated colloids were then dialyzed against
deionized water (DI) until the permeate solution conductivity was
less than 100 mS cm1. The isolated colloids were separated into
two parts. One half was freeze-dried; this is termed HCl-treated
colloids. The other half was dialyzed for at least 24 hours against 4
L of 0.2 M HF to remove silica or silicates and then dialyzed
repeatedly against DI water until the permeate conductivity was
less than 10 mS cm1. This half of the colloids, after HF dialysis,
was freeze-dried and is henceforth referred to as HCl–HF colloids.
A complete description of this separation method and subsequent
characterization of nanoscale material is available elsewhere.41
Control studies were difficult to develop to evaluate potential
artifacts arising from the isolation procedure. Distilled water was
processed through the roto-evaporation step to determine if
atmospheric dust or other contaminants would affect titanium
content of solids. Minimal solids formed and titanium was not
present. Titanium dioxide nanomaterials were observed to be
stable against the dialysis liquids used above at room temperature.
Because titanium has an extremely low solubility, precipitation of
titanium solids from titanium ions during roto-evaporation was
unlikely. However, for other inorganic materials (silver, silica,
etc.) such precipitation concerns would exist. Therefore, the
method employed here may only be suitable for materials such as
titanium dioxide. Overall, the best validation of the roto-evapo-
ration and dialysis method was determined to be a comparison of
titanium content before and after roto-evaporation and dialysis.
Closure of the mass balance could demonstrate that there was no
loss or addition of titanium during the process.
Analytical methods
Because TiO2 has very low solubility,45 Ti in wastewater is
expected to occur solely in solid phases, not in ionic forms. For the
quantification method used in this investigation, these solid
phases must be transformed into ionic forms by acid digestion.
Liquid and solid samples were acid digested using the HNO3/
H2SO4 digestion method for Ti as described by Standard Method
3030 G for water and wastewater analysis.46 The digested samples
were analyzed by Inductively Coupled Plasma Optical Emission
Spectroscopy (ICP-OES) (iCAP 6000 Series, Thermo Scientific,
Cambridge, UK). The method detection limit (MDL) is 0.5 mg Ti
L1 based upon running 10 replicates of 2 and 5 mg Ti L1.47 DOC
was measured by a Shimadzu TOC-VCSH total organic carbon
analyzer. Total dissolved nitrogen (TDN), chemical oxygen
demand (COD), ammonia, nitrate, and nitrite were measured by
HACH DR5000 using the following HACH kits of TNT 828,
method 8000, TNT 831, TNT-836/835, and TNT 839.
Nanoparticle images were obtained using high resolution
transmission electron microscopy (HR-STEM Phillips CM200/
FEG equipped with an energy dispersion X-ray microanalysis
system (EDS)).
Results
Titanium removal at full-scale WWTPs
Samples were collected from the headworks and treated effluent
of ten full-scale WWTPs and analyzed for titanium after acid
digestion. The influent titanium concentration averaged 377 mg

This journal is ª The Royal Society of Chemistry 2011 J. Environ. Monit., 2011, 13, 1195–1203 | 1197
 L1 (median of 321 mg L1; 26 samples) and ranged from 181 to
1233 mg L1. On a percentile distribution, the 90th, 75th, 25th and
10th percentile influent titanium concentrations were 579, 463,
215 and 163 mg L1 (n ¼ 26). Overall, the concentration range is
similar to that which we previously reported for a single plant
sampled several times between 7 am and 5 pm on a single day.48
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We also demonstrated that the morphology of titanium in water
and settled biosolids falls into one of three categories, which were
similar to those used to categorize titanium in foods:48,49 type I—
spheres of TiO2, 100–200 nm in diameter, characterized as the
synthetic food additive polymorph anatase; type II—alumino-
silicates, <100–400 nm in length, generally of flaky appearance,
often with adsorbed surface iron, and mostly characteristic of the
natural clay mineral kaolinite; and type III—mixed environ-
mental silicates without aluminium, 100 to >700 nm in length
and of variable morphology.
Titanium concentrations in treated effluent were significantly
lower than those at the headworks (Table 1). Only two facilities
(#5, #8) had Ti concentrations greater than 10 mg L1, and these
systems achieved nitrification (i.e., most of the total dissolved
nitrogen (TDN) present as nitrate) but not denitrification. It may
be that the denitrification step has biosolids surfaces suitable for
removing titanium bearing colloids or nanoparticles. The other
facilities had low Ti concentrations in treated effluent. Whereas
the titanium concentrations at the headworks exhibited large
variations among duplicate samples, presumably due to variable
loading, sources and size of Ti-bearing materials, the effluent
samples had low variability and usually differed by <10 mg L1
among replicate samples. On a percentile distribution the 90th,
75th, 50th, 25th and 10th percentile effluent titanium concentrations
were 16, 10, 4, 3 and 0.7 mg L1 (n ¼ 30). Although the effluent
concentrations were near our MDL for titanium in water (0.5 mg
L1), treated effluent from microfiltration membrane systems
consistently exhibited the lowest Ti concentrations. Micro-
filtration membrane systems have pore sizes of 100 to a few
hundred nanometres.
Quantification of titanium in colloidal fraction from WWTP
effluents
Because effluent titanium concentrations were low and near the
detection limits of the ICP-OES, a technique was developed to
separate and concentrate colloidal material, including titanium,
from the effluent. Rota-evaporation reduced the sample volume
by approximately 250- to 500-fold. These rota-evaporated
samples underwent dialysis against HCl, or HCl and then HF,
prior to freeze drying. A freeze-dried sample (70 to 200 mg) was
then weighed and acid digested. Titanium concentrations in the
acid matrix ranged from 7.2 to 180 mg L1 (average was 61 mg
L1); duplicate digestions on three samples had less than 15%
variation. These concentrations are well above the detection limit
of the ICP-OES. The mass of HCl-colloids separated from the
water ranged from 11 to 80 mg L1 (average 34 mg L1; ESI,
Table S2‡), as indicated on the x-axis in Fig. 2. The total mass of
HCl-colloids separated from the water, multiplied by the Ti
content of the HCl-colloids after acid digestion, divided by the
initial volume of water from which the colloids were separated,
yields an estimate of the titanium content present in the waste-
water effluent. These values are expressed on the y-axis in Fig. 2.
1198 | J. Environ. Monit., 2011, 13, 1195–1203
Fig. 2 Comparison of titanium content in HCl-colloids relative to the
total colloidal concentration separated from the WWTP effluent.
The trend line in Fig. 2 suggests that effluent samples from across
Arizona with different types of treatment exhibit similar titanium
content per unit colloid mass present in the wastewater effluent.
The slope of the line (95 mg Ti mg1 colloid) provides a reason-
able estimate for the titanium content of WWTP effluents in
Arizona, and the arithmetic average dry mass content of the
HCl-colloids was 66 mg Ti mg1 dry weight of HCl-colloids. The
sample from Facility #1 appeared to be an outlier and is dis-
cussed further in the microscopy section.
While the roto-evaporation and dialysis methods were
primarily designed to obtain samples for electron microscopy
analysis, comparison of titanium content of the isolated samples
to the bulk water samples was considered important to validate
the procedure. Titanium concentrations calculated from the
separation method ranged from 0.2 to 7.1 mg L1 for the WWTP
effluents (Table 1). The difference between the direct measure-
ment of titanium in the treated effluent and the calculation of
titanium content from the separated and acid-digested HCl-
colloids was relatively small. That is, the separation technique
gave the same order of magnitude titanium concentration as the
direct measurement of titanium in the treated effluent. The
membrane bioreactor plants had among the lowest titanium
concentrations. The largest variation was observed in sample
ID#5, which was one of the three samples run in duplicate. The
small discrepancies between the two approaches for obtaining
titanium data can be attributed mainly to obtaining the entire dry
mass of HCl-colloids accurately and assuring complete digestion
of some Ti-containing silicates. Overall, we deemed the rota-
evaporation and dialysis technique acceptable for separating and
concentrating Ti-bearing colloids from large volume and dilute
water samples.
Electron microscopy characterization of colloids from WWTP
effluents
Electron microscopy was conducted on most of the colloidal
samples, including samples from the same location both before
and after HCl or subsequent HF treatment. Select samples are
presented that represent the overall trends observed. HCl
This journal is ª The Royal Society of Chemistry 2011
 treatment removes most of the carbonates and hydroxides,
whereas additional HF–HCl treatment dissolves most of the
silicates. The most significant finding is that TiOx nanoparticles
are present in treated wastewater effluent. Clusters of TiOx
nanoparticles in the HCl–HF treated colloid WWTP effluent
sample from Facility #2 are shown in Fig. 3. A high resolution
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with diameters of approximately 10 nm that have crystalline
lattices. EDS of the cluster indicated the presence of primarily Ti,
O, Cu and C, with trace amounts of other elements. Copper and
carbon are present in the TEM grid. Carbon, and some oxygen, is
present in the sample as colloidal organic material.41 The crys-
talline lattice, dominant Ti and O peaks, and lack of silica

TEM image of the cluster indicates the presence of nanoparticles support the conclusion that TiOx nanoparticles are present in the

Fig. 3 TEM images of (A) clusters of nanoparticles in the HCl–HF treated colloid WWTP effluent sample from Facility #2; (B) high resolution TEM
image of the cluster indicates the presence of nanoparticles with diameters of approximately 10 nm that have crystalline lattices (inside circles); (C) EDS
of cluster indicating elemental composition.

This journal is ª The Royal Society of Chemistry 2011 J. Environ. Monit., 2011, 13, 1195–1203 | 1199
 sample. The TiOx nanoparticles also appear to be fused together;
this could represent the morphology of the TiOx as used in
society or may have occurred during colloid isolation and
preparation.
TiOx nanoparticles from the same site (Facility #2) could not
be identified in HCl-colloid samples. On the basis of EDS anal-
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ysis, the HCl-treated samples contained high concentrations of

silicon dioxide nanoparticles, which formed large clusters of
discrete nanoparticles (Fig. 4); titanium was present in these
samples but at very low elemental mass ratios due to the large
amount of silica present. High resolution TEM indicates that the
discrete nanoparticles were approximately 20 to 30 nm in
diameter (Fig. 4B), larger than the TiOx nanoparticles observed
after HCl–HF treatment (Fig. 3). Fig. 4B indicates that the
nanoparticles are amorphous, unlike the more crystalline TiOx
nanoparticles shown in Fig. 3B. The amorphous silica-based
nanoparticles may be present in the WWTP effluent or may form
during colloid isolation. In the HCl-treated sample from Facility
#1 (not shown) silica particles were non-spherical structures,
rather elongated and planar with sharp edges. Additional
research is underway to address the presence of silica nano-
structures, but is important to note because titanium is often
substituted for silica with silicate minerals.
A few additional examples of the different types of titanium
structures present after HCl–HF treatment are illustrated in
Fig. 5. Samples from Facility #8, which contained the second
largest mass concentration of titanium in the WWTP effluent
(Table 1), indicate the presence of very small (4 to 5 nm
diameter) nanoparticles (dark spheres) embedded in an amor-
phous matrix of colloidal organic matter and some silica. High
resolution TEM could not determine if these small spheres were
crystalline, but EDS confirmed high levels of titanium and
oxygen with lower amounts of carbon, copper and silica. The
high abundance of TiOx nanoparticles in samples from Facility
#8 meant that they could be imaged after only HCl treatment
(not shown); larger crystalline structures (20 to 30 nm) were
observed in these images. In addition, larger titanium-containing
particles were found in HCl-treated samples from Facility #6.
These particles were aggregated with silica nanoparticles in large
clusters with sizes of several micrometres, as illustrated in
Fig. 5B. Based on EDS analysis (not shown), the particles were
composed of iron phosphate in addition to magnesium and
titanium. Samples from Facility #1 differed from those from
Fig. 4 TEM images showing (A) primarily silicon dioxide nanoparticles
in HCl treated sample from Facility #3 (B) high resolution image
showing amorphous minerals.
Fig. 5 TEM images of samples after HCl–HF treatment from (A)
Facility #8; (B) Facility #6; (C) Facility #1; (D) high resolution image of
the circled region in (C) in which the dashed circle shows the presence of
Ti-based nanoparticles that appear embedded in a larger mineral.
Facilities #3, #6 or #8 described above. TiOx nanoparticles with
sizes of 4 to 5 nm were present in HCl–HF treated colloids from
Facility #1. However, these nanoparticles (Fig. 5D) appeared to
be encased in large particles containing iron, aluminium, chlo-
ride, phosphate, and sulfate (Fig. 5C). These mineral differences
as well as the differences in silica morphology noted above in the
Ti-bearing material from Facility #1 may shed light on why this
site is somewhat of an outlier in Fig. 2.
Discussion
Removal of the colloid-sized material in treated wastewater is
affected by the design and operational efficiency of each unit
process (sedimentation, granular media and/or membrane sepa-
ration). Previous mass flow modeling of nanomaterials during
wastewater treatment considered fixed removal efficiencies (97%)
of particles based solely on their size <100 nm or uniform
removal distributions (90.6% to 99.5%) as determined by bench-
scale studies in the presence of biomass.14,50 Based on Table 1, the
overall removal efficiency of titanium is quite high (96.1% to
99.4% with an average removal of 98.3%). However, previous
research in which we conducted size fractionation of titanium
across one of the facilities included in this paper indicated quite
low removal (<30%) of titanium that passed through a 0.7 mm
nominal size glass fiber filter.18 Several observations may
reconcile these apparent differences.
First, not all titanium present in wastewater is of a nano- or
colloidal size of <100 nm. In fact, as Fig. 3–5 indicate, the TiOx
nanoparticles we detect in wastewater effluents appear to be 4 to
30 nm in diameter. Typical WWTP sedimentation processes are
designed with surface loading rates on the order of 30 m per day.
On the basis of discrete particle settling (i.e., Stokes settling

1200 | J. Environ. Monit., 2011, 13, 1195–1203 This journal is ª The Royal Society of Chemistry 2011
 velocities), these loading rates are suitable only for removing
particles larger than 5 to 10 mm and particles with densities of 1.5
to 4.5 g cm3 at 20  C; TiO2 has a density of 4.2 g cm3. Thus
sedimentation (primary or secondary, Fig. 1) cannot remove
discrete and stable NPs with sizes <0.1 mm. Titanium-containing
particles (e.g., clays), aggregates and clusters of TiO2, or TiO2
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sorbed to biomass could be large enough to settle. Actual
removals and minimal size removed are difficult to calculate due
to a process called flocculant settling in which aggregating
particles continually change in size, shape, and specific gravity.51
Thus the size distribution of titanium in WWTP influent and
across WWTP unit processes strongly influences its potential for
removal by sedimentation processes.
Second, biological treatment processes designed to decompose
domestic and industrial compounds through metabolic processes
and cellular growth can affect nanoparticle removals. Two basic
means of biological treatment were included in our sampling
campaign. Suspended biomass in activated sludge processes was
the most common, whereas attached biofilms were used only at
Facility #8, which employed trickling filters. Diffusion governs
the movement of nanoscale particles, including nano-TiO2, near
and within biofilms and suspended biomass.16 Many types of
nanoparticles, including TiO2, readily sorb to suspended biomass
from WWTPs.33 Once attached, the removal of the NP becomes
connected to the management and removal of the suspended
biomass. WWTP secondary sedimentation unit processes are
very effective at settling biomass and thus would remove sorbed
NPs. This explains the high removals reported in the batch
experiments that served as the basis for mass flow modeling of
NPs.14,22 Furthermore, NPs undoubtedly sorb to attached
biomass (i.e., biofilms in trickling filters), although no data
specifically related to WWTPs are yet available. However, filter
media coated with biofilm extracellular polymers retained more
NPs than uncoated filter media to an extent unaccountable for
simply by electrostatic attraction.52 Carboyxlated and PEG-
coated quantum dots also accumulated in biofilms.53 In the
presence of high concentrations of NPs, biofilm sloughing due to
silver NPs was observed, whereas carbon nanotubes affect bio-
film attachment and exhibit potential NP–biofilm
interactions.54,55
Third, although filtration processes are common in some
European countries, most WWTPs in the USA do not include
them.50 However, submerged and pressurized microfiltration
membrane systems are becoming an increasingly common means
of achieving secondary solids separation (Fig. 1) and are part of
two sampled facilities (Table 1). These membranes commonly
have 0.1 to 0.4 mm pore sizes and achieve very high levels of colloid
removal.56 Although the beneficial use of nanoparticles in
conjunction with innovate membrane treatment systems has been
identified, few reports specifically on rejection (i.e., removal) of
engineered nanoparticles from wastewater by membranes
currently exist.24,25,57–60 In one study, ultrafiltration membranes
were observed to remove polystyrene or magnetic nanoparticles
(20 to 250 nm) better than microfiltration membranes did. The
removal mechanisms for colloids and membranes are complex
and involve not only size exclusion but also colloid surface charge
interactions with bare membrane or membrane foulants; the
mechanisms are also affected by water movement (dead-end
versus cross-flow membrane designs). Based on the TSS, COD and
This journal is ª The Royal Society of Chemistry 2011
NP data presented in Table 1, the membrane bioreactor facilities
(#9 and #10) achieved among the highest water quality. These
facilities use microfiltration membranes to separate activated
sludge suspended biomass from the water. In contrast, Facility #1
processes secondary settled water (biomass separation process)
through microfiltration as a pretreatment for reverse osmosis. The
microfiltration removes greater than 60% of the titanium-bearing
nanoparticles present in the secondary settled water (Table 1).
Overall, micro- or ultrafiltration membrane processes appear
capable of achieving the highest levels of NP removal as compared
with trickling filter or activated sludge systems with conventional
gravity secondary sedimentation.
Summary and conclusions
Titanium oxide (TiOx) nanoparticles were detected in waste-
water effluents from 10 municipal facilities at concentrations
ranging from <2 to 20 mg L1, which is consistent with our
previously reported findings from a single WWTP.18 The
biological wastewater treatment facilities removed, on
average, 98.3% of the incoming titanium, although the size
and composition of materials containing titanium in the
influent wastewater sewage was not characterized. Attached
(trickling filters) and suspended (activated sludge) biological
treatment processes play an important role in ‘‘trapping’’ NPs
in biomass, which can then be settled or removed via
membrane filtration. This paper is the first to identify TiOx
nanoparticles in WWTP effluents by high resolution TEM,
which indicates that they are 4 to 30 nm in diameter and
roughly spherical. WWTP effluents commonly discharge
treated water into lakes, rivers and streams, and this work
clearly documents that the release of nanoscale TiOx into the
environment is possible. It appears that microfiltration-
treated wastewaters contain fewer nanoparticles than
conventionally settled wastewater, and this additional benefit
to the use of membranes in wastewater treatment should be
explored. Although beyond the scope of this titanium-focused
paper, our findings suggest the presence of silica nanoparticles
in wastewater effluents at far higher concentrations than
TiOx. Our study was not designed to quantify silica nano-
particles, which would require working with non-silica-based
sampling equipment, filters, and separation techniques that
could avoid potential precipitation of silica colloids, but this
is the focus of ongoing research.
Acknowledgements
Although the research described in the present study has been
funded in part by the U.S. Environmental Protection Agency
through a grant/cooperative agreement (RD833322), it has not
been subjected to the Agency’s required peer and policy
review. Therefore, it does not necessarily reflect the views of
the Agency, and no official endorsement should be inferred.
Additional funding was provided by the Paul L. Busch
Research Award from the Water Environment Research
Foundation. Characterization of nanoparticles was partially
supported by the NIH Grand Opportunities (RC2) program
through NIEHS grant DE-FG02-08ER64613 and the use of
facilities within the LeRoy Eyring Center for Solid State
J. Environ. Monit., 2011, 13, 1195–1203 | 1201
 Science and the Goldwater Environmental Laboratory at
Arizona State University. Alex Weir and Yifei Wang, graduate
students at ASU, assisted in sample collection and digestion.
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