Journal of Hazardous Materials 463 (2024) 132938

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Journal of Hazardous Materials

journal homepage: www.elsevier.com/locate/jhazmat

Fate and potential risks of microplastic fibers and fragments in water and
wastewater treatment processes
Sang-Heon Na a, b, 1, Min-Ji Kim a, 1, 2, Jihee Kim a, Rida Batool a, b, Kyungjin Cho a, b,
Jaeshik Chung a, b, Seunghak Lee a, b, Eun-Ju Kim a, b, *
a
Water Cycle Research Center, Korea Institute of Science and Technology (KIST), Seoul 02792, South Korea
b
Division of Energy and Environment Technology, KIST School, Korea University of Science and Technology, Seoul 02792, South Korea

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• The properties of MPs had great impact

on their removal along treatment
process.
• Sand filter showed higher MP removal
than activated sludge process or
coagulation.
• In total, about 40 % of MPs was retained
in sludge during activated sludge
process.
• MP-leached DOMs under UV/HP and
NaClO treatments exhibited distinct
toxicity.

A R T I C L E I N F O A B S T R A C T

Editor: Joao Pinto da Costa Water and wastewater treatment plants (WWTPs) receive various types of microplastics (MPs), with fibers and
fragments being dominant shapes. Here we investigated the removal behavior and transformation of MPs
Keywords: (polypropylene and polyethylene terephthalate fibers and fragments) in simulated water and wastewater
Microplastics treatment units, including activated sludge process, coagulation, sand filtration, and advanced oxidation/
Removal
disinfection. Sand filtration demonstrated the highest average efficiency in removing MPs (98 %), followed by
Sludge
activated sludge process (61 %) and coagulation (55 %), which was associated with their physicochemical
Leaching
Cytotoxicity properties (shape, size, density, surface functional groups, etc). In activated sludge process and coagulation, the
polymer type had a greater impact on the removal of MPs than the particle shape, while in sand filtration, the
particle shape played a more important role. When subjected to the long-term operation and backwashing of
sand filters, approximately 15 % of the initially filtered fragments broke through the sand media, with nearly no
fibers escaping. UV-based advanced oxidation and chlorination induced the leaching of dissolved organic matters
with different molecular characteristics from fragment MPs, resulting in varying levels of cytotoxicity and
bacterial toxicity. Our study provides important information for predicting the fate of MPs and mitigating their
impacts in WWTPs.

* Corresponding author at: Water Cycle Research Center, Korea Institute of Science and Technology (KIST), Seoul 02792, South Korea.
E-mail address: eunjukim@kist.re.kr (E.-J. Kim).
1
These authors contributed equally to this work
2
Present address: Infra Engineering Group, Samsung Electronics Co. Ltd., Suwon 16677, South Korea

https://doi.org/10.1016/j.jhazmat.2023.132938
Received 20 July 2023; Received in revised form 9 October 2023; Accepted 3 November 2023
Available online 7 November 2023
0304-3894/© 2023 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY-NC license (http://creativecommons.org/licenses/by-
nc/4.0/).
S.-H. Na et al.
1. Introduction
An exponential increase in mass production and use of plastics had
led to the widespread proliferation of small plastic debris, which is
defined as microplastics (MPs, <5 mm), in various environments [15].
MPs are present in seawater, freshwater, rain droplets, air, and soil [10,
17,27,39]. One of the primary sources of MPs in the environment is
through treated effluents from water and wastewater treatment plants
(WWTPs) [34]. Although existing WWTPs are not specifically designed
to remove them, a significant amount of MPs can be removed, with a
removal rate of 60–99 % depending on the treatment process employed
[14,16,2,28]. Despite the growing number of studies on the fate and
transport of MPs in WWTPs, most of them focus on individual treatment
processes. Still, we do not have a comprehensive understanding of how
MPs are affected by and transformed as they are transported through
different successional treatment compartments within WWTPs. This
knowledge gap introduces uncertainty in predicting the environmental
and health impacts of MPs after being discharged from WWTPs into
drinking and natural waters.
Advanced oxidation processes (AOPs) and chlorine disinfection as a
tertiary treatment process at the end of WWTPs, may alter the intrinsic
properties of MPs such as morphology, surface roughness, hydrophilic­
ity, and oxygen content [29], thereby affecting the leaching of various
organics [13,21,24,33]. The organic leaching from MPs during disin­
fection and AOPs can pose adverse effects on the environment and biota
because a wide range of toxic chemicals are often incorporated into
plastics. Across aquatic organisms, some consistent results have been
observed, for example, the leaching fraction from MPs showed higher
toxicity compared to MPs themselves [11,31,7]; however, most of these
studies were conducted under artificially accelerated treatment condi­
tions (e.g., high oxidant dose and long irradiation time). Due to the
limited information available on the characteristics of dissolved organic
matter (DOM) leached from MPs under normal operation conditions
applied in WWTPs, it remains unclear whether the same negative effect
would be observed for MPs-derived DOM.
The present study aims to investigate the transport and fate of MPs in
lab-scale treatment systems, including activated sludge process, coagu­
lation, rapid sand filtration (RSF), and UV-based AOP/chlorine disin­
fection, all of which are employed in conventional WWTPs [12]. While
the shapes and polymer types of MPs found in water and wastewater are
diverse, for the sake of simplicity we focus our study on fragments and
fibers composed of polypropylene (PP) and polyethylene terephthalate
(PET) due to their relatively high proportions [22,43]. Additionally, we
examine the leaching behavior of MPs during AOPs and disinfection
over the technologically relevant time scale (60 min), followed by
toxicity assessments of MPs-derived leachates using human cell line and
microbioassay. Therefore, our study fills an important knowledge gap in
the understanding of the behavior of MPs in WWTPs.
2. Material and methods
2.1. Chemicals and materials
The chemicals used in the study were: polyaluminum chloride (PAC;
7.6 ± 0.3 % Al3+), sodium hypochlorite (NaClO, Sigma-Aldrich), and
hydrogen peroxide (H2O2, Merck). To investigate the behavior of MPs
under realistic conditions, wastewater samples were taken in the
effluent of primary clarifier, influent and effluent of secondary clarifier,
and effluent of sand filter in a municipal wastewater treatment plant,
South Korea (Fig. S1), and were filtered through a glass filter (1 µm pore
size; Pall Life Sciences). For each wastewater sample, pH, total sus­
pended solids (TSS), hardness, and dissolved organic carbon (DOC) were
analyzed, as shown in Table S1. In this study, we prepared MPs using
PET (23 µm diameter, Goodfellow ES30-FB-000130) and PP (28 µm
diameter, Goodfellow PP30-FB-000147) filaments according to the
cryotome method [6]. Briefly, the PET and PP filaments were coated
2
Journal of Hazardous Materials 463 (2024) 132938
with FSC 22 frozen section compound (Leica) and frozen at – 80 ◦ C. The
frozen filaments were then cut into 50 and 200 µm in lengths using a
cryogenic microtome (Leica CM 1800). Sections were washed with
deionized (DI) water at 60 ◦ C, filtered with a 0.45-µm polycarbonate
filter (Millipore), and stored suspended in DI water as appropriate.
Sections with an aspect (length-to-diameter) ratio < 3:1 or > 3:1 were
denoted as fragments or fibers, respectively [6]. The morphologies and
sizes of MPs were determined using a scanning electron microscopy
(SEM; Hitachi Regulus 8230). The compositions of MPs were detected by
Fourier transform infrared microscope (FTIR microscope; Bruker
LumosII) in attenuated total reflection (ATR) mode. The characteriza­
tion results are provided in the Supporting Information (Fig. S2).
2.2. Activated sludge reactor setup and operation
Lab-scale activated sludge reactors with a working volume of 1 L
were operated at room temperature. The reactors were equipped with an
air pump to keep a dissolved oxygen concentration above 3.5 mg/L, and
magnetic stirrer for mixing. To achieve a mixed liquor suspended solids
(MLSS) concentration of 2000 mg/L, seeding sludge collected from an
aerobic tank in a full-scale wastewater treatment plant was suspended in
filtered effluent of primary clarifier. MPs were spiked into the reactors to
reach a final concentration of approximately 1000 particles/L. The
bioreactor operated for 3 h at a drainage ratio of 50 %, which comprised
0.2 h feeding, 2.5 h aerobic reaction, 0.15 h settling, and 0.15 h effluent
withdrawal. To measure the concentration of MPs, the supernatant was
collected at the end of the operation. After a series of extraction pro­
cesses involving organic digestion and density separation, the concen­
tration of MPs in the supernatant was quantified by FTIR microscope
(Bruker LumosII).
2.3. Coagulation experiments
Coagulation experiments were conducted in a 1-L jar tester using the
secondary clarifier influent at a concentration of ~1500 particles/L. The
coagulant was added into the working solution, followed by rapid stir­
ring at 160 rpm for 2 min, slow stirring at 60 rpm for 30 min, and was
then allowed to settle for 10 min. The supernatant and subnatant were
used to quantify the residual PP and PET MPs, respectively, based on the
difference in their densities. The collected samples were filtered through
5-μm polycarbonate membrane black filters (SterliTech), and MPs on the
filters were counted using FTIR microscope (Bruker LumosII). The
freeze-dried coagulation flocs were also examined using an SEM
equipped with energy-dispersive X-ray spectroscopy (SEM-EDX; Hitachi
Regulus 8230) and X-ray photoelectron spectroscopy (XPS).
2.4. Sand column experiments
The RSF process was simulated through column experiments. Sand
(effective grain size ranging 0.3–0.45 mm and a uniformity coefficient <
2) was wet-packed into the cylindrical glass columns (5 cm length and
2.5 cm inner diameter). Two stainless steel screens with a pore size of
250 µm were used to seal each of the columns at the inlet and outlet. The
column was operated in down flow mode at a filtration rate of 4.6 m/h.
Backwashing was performed with water only, at a backwash rate of 30
m/h. Prior to each experiment, the column was equilibrated to the
steady condition by flushing 20 pore volumes (PVs) of secondary clari­
fier effluent. 3 PVs of plastic suspension (~2000 particles/L) were
subsequently injected into the columns followed by secondary clarifier
effluent for 2 PVs. The sand filter effluents were filtered through 5-μm
polycarbonate membrane black filters (SterliTech), and the number of
MPs on the membrane was determined by FTIR microscope (Bruker
LumosII). Each column was imaged using X-ray Computed Tomography
(XCT, Bruker Skyscan1172) to visualize the spatial distribution of MPs
within the sand pack.
S.-H. Na et al.
2.5. UV/H2O2 advanced oxidation and chlorination experiments
All UV/hydrogen peroxide (UV/HP) oxidation and chlorination ex­
periments were carried out using the effluent after sand filtration. For
UV/HP oxidation, 50 µm PP and PET suspensions (~2000 particles/L)
and 5 mM HP were applied into a cylindrical quartz reactor and stirred
under UV light for 1 h. The light was illuminated with a 300 W Xe arc
lamp (Newport, 66902). For chlorination, MPs (~2000 particles/L)
were exposed to sodium hypochlorite solution (NaClO, 5 mg/L free
chlorine) for 1 h at a CT value of 300 mg min L− 1 consistent with that
expected for WWTP conditions [37]. Sodium thiosulfate solution (100
mM) was used to quench residual H2O2 and NaClO in the samples. The
collected suspensions were filtered with a 0.22-μm membrane for
further chemical analysis and toxicity testing.
The fluorescence characteristics of the DOM leached from UV/HP or
chlorination processes in the filtrate were analyzed by excitation-
emission matrix (EEM) fluorescence spectrophotometer (Shimadzu RF-
6000). The low molecular weight organic compounds present in the
filtrate were extracted with solid-phase extraction (SPE) cartridges (HLB
Plus Oasis, Waters) and identified on gas chromatography coupled to
mass spectrometry (GC-MS, Agilent 6890 N with LECO PEGASUS MSD).
Details of SPE protocol and analytical conditions can be found in the
Supplementary Information. The toxicity of weathered MPs and leach­
ates was assessed using human dermal fibroblast (HDF) cells and the
ToxiChromo Test kit (EBPI). Cytotoxicity of the samples in HDF cells was
measured by the 3-[4,5-dimethylthiazol-2-yl]- 2,5-diphenyl tetrazolium
bromide (MTT) assay (Sigma-Aldrich). The Toxi-ChromoTest kit was
used according to the manufacturer’s instructions to determine the
samples’ potential for the inhibition of de novo synthesis of an inducible
enzyme, β-galactosidase, in a mutant Escherichia coli [23]. Toxicity as­
says are detailed in the Supplementary Information.
2.6. Statistical analysis
Data were presented as mean ± standard deviation (S.D.). One-way
ANOVA with Tukey’s HSD test (IBM SPSS Statistics 20) was performed
to identify statistically significant differences (p < 0.05) between the
leachate samples in toxicity assays.
3. Results and discussion
3.1. Removal of fragment and fiber MPs in activated sludge reactor
The analysis of MPs in the effluent fractions of bioreactors showed
that on average, 61 % of MPs initially added were removed from the
water phase (Fig. 1). Since the microbial degradation of MPs is expected
to be negligible with such a short hydraulic retention time, this suggests
that a significant portion of MPs was accumulated in the settled sludge.
However, it should be noted that the removal efficiency of MPs in the
activated sludge-based process may vary with operational parameters
such as hydraulic retention time, solid retention time, and MLSS con­
centration [32], and this should be considered in future studies. The
considerable removal of low-density PP particles from the bioreactor
effluent indicated the involvement of additional removal mechanisms,
such as interactions between MPs and extracellular polymeric sub­
stances (EPS) in the sludge, besides density. In this context, the higher
removal efficiency of PET (78 %) compared to PP (46 %), as shown in
Fig. 1, may be linked to the enhanced adsorption of EPS onto PET, due to
the presence of oxygen-containing functional groups on the surface of
PET [41]. As for the morphology, fibers were removed from the water
more effectively than fragments (Fig. 1), which could be attributed to
their greater tendency to entangle with sludge flocs and settle [26]. This
finding is consistent with previous studies that the majority of MPs
present in the secondary sludge are fibers [1,30]. From the results ob­
tained in activated sludge reactor experiments, we confirmed that the
secondary sludge serves as an important sink for MPs removed from
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Journal of Hazardous Materials 463 (2024) 132938
wastewater in wastewater treatment plants.
3.2. Removal of fragment and fiber MPs in coagulation
The efficiency of MP removal was explored with different doses of
PAC (1–10 mg Al/L) at pH 6.5. Overall, the removal efficiencies for PET
were higher than those for PP, irrespective of their shape (Fig. 2). PET
removal was substantially enhanced even at the lowest PAC dose (1 mg/
L) and then did not further increase at doses greater than 1 mg/L, while
the efficiency of PP removal gradually increased as the PAC dose
increased from 1 to 10 mg/L. The difference in the removal efficiency
for PET and PP during coagulation may be ascribed to a combination of
factors, including density, the amount of charge, and the binding affinity
between the coagulant and MPs. Indeed, PET had more negative zeta (ζ)
potential values compared to PP (Fig. S3).
Moving forward, we conducted isothermal titration calorimetry
(ITC) measurements to identify the type of interaction and quantify the
binding affinity between PAC and PP or PET fragments. Fragments were
used for ITC experiments to avoid settling issues encountered when
applying fibers. Thermograms resulting from the titration of PAC into
suspensions of PP and PET fragments displayed strong, positive peaks
implying that the coagulation process proceeds via an endothermic
pathway (Fig. 3). In both cases, the interaction between MPs and PAC
was entropy-driven and thermodynamically favored as demonstrated by
the negative Gibbs free energy and positive entropy. The significantly
larger entropic contribution to the overall change in Gibbs free energy
indicates the involvement of desolvation in MP coagulation. Unlike
relatively similar thermodynamic parameters, the binding constant (Ka)
for PET (9.36 ± 0.06 × 102 M− 1) was approximately 3.6 times greater
than that of PP (2.59 ± 0.02 × 102 M− 1). This confirms that high PET
removal efficiency is linked to the binding affinity between PAC and
PET, likely due to the presence of C– – O bond within the PET molecule
enabling additional binding of Al species [3].
To reveal the coagulation mechanism, we first measured the ζ po­
tential of MPs in the presence of PAC. The ζ values were not significantly
affected by PAC doses in the range of 0–5 mg/L, and charge inversion
from negative to positive potentials was only observed at a PAC dose of
10 mg/L (Fig. S3). This suggests that in addition to charge neutraliza­
tion, other mechanisms should be responsible for MP coagulation by
PAC. Given that the hydrolysis products of PAC comprise a series of Al
species contributing to different mechanisms and efficiencies in coagu­
lation, the composition of Al species in flocs was analyzed by XPS.
Fig. S4 shows the Al 2p spectra of freeze-dried flocs. The Al 2p peak was
split into two overlapping bands associated with tetrahedral aluminum
(AlIV) and octahedral aluminum (AlVI) [42]. The AlVI/AlIV ratios for all
Fig. 1. Remaining MP fraction in bioreactor effluent. [MPs]0 = 1000 parti­
cles/L.
S.-H. Na et al.
Fig. 2. Effect of PAC concentrations on the removal of PP and PET fragments
and fibers. [MPs]0 = 1500 particles/L.
flocs were less than the theoretical value in Al13 molecules, 12, [20],
indicating the presence of Al13 aggregates in the precipitates. The for­
mation of Al13 aggregates with dense structure was further verified by
SEM-EDS (Fig. S5). Taken together, at a given pH, the coagulation of
MPs was expected to undergo charge neutralization by preformed Al13
and then interparticle bridging by Al13 aggregates.
3.3. Retention of fragment and fiber MPs in RSFs
The polymer type had less influence on the transport characteristics
of MPs, given similar breakthrough curves for PP and PET fragments
(Fig. S6); thus, PET was selected to examine the retention of MPs in
porous media. The recoveries of PET fiber and PET fragment were 98.5
± 2.3 % and 97.4 ± 1.6 %, respectively, showing their high retention in
the sand column. These very low recovery values were likely size
limited. PET fragment (23 µm × 50 µm) and fiber (23 µm × 200 µm)
dimensions are in the same range as the average pore-throat size in our
system (~40 µm). On the other hand, the irregular shape of MPs may
significantly affect the high retention of PET fragments and fibers in the
porous media. Irregularly shaped MPs with high aspect ratios are placed
at random orientation when advected [25], allowing them to drift across
the fluid streamlines. Consequently, the rotation of irregular MPs, aided
Fig. 3. ITC integrated heat data for the interaction of PAC and (a) PP and PET fragments (Inset: extracted thermodynamic parameters). The solid line represents the
best fit for a single site model.
4
Journal of Hazardous Materials 463 (2024) 132938
by drift due to fluid hydrodynamics, can lead to a large increase in the
probability of particle attachment, which further explains the observed
straining effects. The lower mobility of nonspherical colloids compared
to spherical colloids in porous media has been noted previously [36,40].
Moving forward, we employed XCT to directly visualize the vertical
distribution of MPs in sand packs. PET fragments reached the maximum
depth of ~4.5 cm and PET fibers infiltrated to the minimum depth of
~1.5 cm (Fig. 4a), indicating that more fibers were retained at the up­
stream collector surface relative to downstream. The prominent differ­
ence in depths at which fiber and fragment MPs were detected also
reflects the important role of particle shape in their transport, rather
than surface properties such as surface charge, since they had similar
ζ-potentials (–5.6 ± 0.4 mV for fragments and –5.1 ± 0.7 mV for
fibers).
In practice, the RSFs are operated for longer times and require
backwashing. We thus tested the effects of long-term filtration and
backwashing on the release of MPs from the sand media. Unlike a sig­
nificant release of fragments, fibers were released only in low pro­
portions (~2.5 %) over longer filter run times (Fig. 4b), likely due to
physical constraints. Additionally, during backwashing, 1–5 % of MPs
captured were released into the backwash water (Fig. 4b). This implies
that the backwashing had no discernible impact on the retention of MPs
in RSFs, despite using a typical backwashing mode at a flow rate of
30 m/h [4]. Together, these results demonstrate that the RSF used in
tertiary treatment offers a high efficiency in removing MPs.
3.4. Leaching of DOM from fragment MPs during UV-AOP and
chlorination
Although fibers and fragments may exhibit different stabilities as
subjected to photochemical forces [38], fragments were chosen for
UV-based oxidation and chlorination experiments due to their greater
breakthrough from saturated porous media. Having demonstrated that
UV-based AOPs and chlorine disinfection, commonly employed in
WWTPs, could lead to the release of organic matters from MPs due to
irreversible alterations and damage to MPs [19,29,9], we elected to
explore how UV/HP and chlorination processes influence the charac­
teristic of MP leachates over a technically relevant time frame (1 h).
Fig. S7 shows the levels of DOC and SUVA254 for the leachates. As ex­
pected, PP and PET released significantly higher amounts of DOCs after
being treated with UV/HP and NaClO than their corresponding dark
treatments (Fig. S7a). However, DOC leaching exhibited no significant
difference across treatments regardless of the polymer type (p > 0.05,
S.-H. Na et al. Journal of Hazardous Materials 463 (2024) 132938

Fig. 4. (a) On the left: XCT images for 5-cm sand columns with PET fibers (top) and PET fragments (bottom). On the right: representative cross-sectional XCT images
taken at the top and bottom layers of sand columns (indicated by red lines), respectively. The MPs are indicated by red circles. All scale bars represent 200 µm. (b)
Filtration efficiency of MPs in RSFs before and after backwashing.

8.6–9.9 mg/L for NaClO treatment vs. 8.2–9.0 mg/L for UV/HP treat­
ment), suggesting that the treatment method did not impact the extent
of DOC leaching. In addition, SUVA254 for the PP-derived DOM was
substantially smaller than that for the PET-derived DOM (Fig. S7b),
which is attributed to the absence of aromatic moieties in the backbone
of PP. Despite similar DOC concentrations, SUVA254 value in the
UV/HP-treated PET was higher than that in the NaClO-treated PET. This
indicated that the characteristics (i.e., component and molecular
weight) of DOMs leached from MPs differed between UV/HP and NaClO
treatments due to the higher reactivity of MPs with •OH than with HClO.
Fluorescent EEMs of leachates from PET and PP fragments under
different treatment conditions were compared to further characterize
the possible components of MPs-derived DOM. The properties of fluo­
rescent DOM leached from MPs showed that protein and phenol-like
fluorescence regions (i.e., regions I, II, and IV), such as peaks B, N,
and T, were predominant in all samples (Fig. 5a). This is likely due to the

Fig. 5. (a) Fluorescence EEM spectra of the DOM leached from PET and PP in the dark and after UV/HP or NaClO treatment. (b) Heatmap showing 52 intermediates
(identified by GC-MS) in the leachates obtained after UV/HP or NaClO treatment. Color key indicates the presence of the intermediates. C1 and C2 represent PP and
PET dark controls; S1, S2, S3, and S4 represent UV/HP-treated PP, NaClO-treated PP, NaClO-treated PET, and UV/HP-treated PET samples.

5
S.-H. Na et al.
presence of many fluorescent compounds in the sand filtration effluent,
which is the background wastewater. Notably, the fluorescence in­
tensities of humic-like component (region V) increased from 1.1 to 2.4
times in DOM leached from NaClO- and UV/HP-treated MPs along with
the increase of peak B intensity compared to their dark controls (Fig. 5a).
These results imply that the fluorescence feature of MP-derived DOM
could be influenced by UV/HP and chlorination under conditions that
closely mimic actual process conditions.
Confident that both treatments enabled the generation of low mo­
lecular weight fractions from PET and PP fragments, we hence applied
non-targeted GC-MS analysis for the profiling of low molecular weight
organic compounds in the leachates. It should be noted that, although
we were able to identify a large number of organic compounds present in
leachates, they represent only a small fraction of the total leachate. Out
of 40 compounds that were identified only in the leachates, 3 com­
pounds (7.5 %) co-occurred. The differences in polymer structure of PET
and PP, in combination with the presence of different plastic additives
[8], may give rise to the differences in the chemical composition across
the leachate samples that we observed (Fig. 5b). The total number of
compounds in a given sample was highest for UV/HP-treated PP and
lowest for NaClO-treated PP (Fig. 5b). Principle component analysis
revealed that the organic profile of UV/HP-PET was clearly distinct from
that of other samples, whereas the organic profiles from NaClO-PP and
NaClO-PET were similar (Fig. 5b). Some compounds such as 9-octadece­
namide, atrolactic acid, acetic acid ethyl ester, 4-(4-Acetylphe­
nyl)-4-hydroxy-2-methyl-1-pentene, 3-Methoxy-3-phenyl-1-propyne,
dimethylacetal dodecanal, and 1-chloroheptane were found only in the
UV/HP-treated PET. Also, the NaClO-treated samples contained 12
specific compounds, including disinfection byproducts (DBPs). The
DOM leached from MPs has been shown to cause the formation of DBPs
upon chlorination [24]. However, the DBPs (trichloromethane and
1-chloro-2-propanone) appeared only in the NaClO-treated PET, likely
due to the difference in the chlorine resistance of the two polymers. PP
has been reported to be more resistant to chlorine compared to PET [5].
These results demonstrated that the chemical profiles of MPs-derived
DOM are dynamic, and vary depending on the treatment method and
polymer type.
To determine whether a relationship exists between the dynamics of
MP leachate composition and their toxic effects, we tested the toxicity of
leachates using a battery of cell-based assays. The acute cytotoxicity of
Fig. 6. (a) Viability of HDF cells following 24-h exposure with the leachates obtained from dark or UV/HP and NaClO treatments of PP and PET. Cell viability data
are presented as a percentage relative to the non-treated control cells. (b) Biotoxicity of leachates obtained from dark or UV/HP and NaClO treatments of PP and PET
in terms of percent inhibition of enzyme biosynthesis. Averages and standard deviations are shown (n = 5); different letters above the bars indicate significant
differences according to One-way ANOVA (p < 0.05).
6
Journal of Hazardous Materials 463 (2024) 132938
the weathered MPs and their leachable fractions treated by UV/HP and
NaClO was assessed in HDF cells. The weathered MPs were not found to
be cytotoxic (Fig. S8), which is consistent with previous studies [18,35].
For the leachates, while both dark controls and NaClO-treated samples
showed no cytotoxicity, the cell viability in the UV/HP-treated samples
was significantly reduced to 50–70 % (Fig. 6a). Conversely, in the
microbioassay (Toxi-Chromo test), we observed a more pronounced
inhibition of β-galactosidase biosynthesis in the NaClO-treated leachates
(~95 %) than in the UV/HP-treated leachates (55–70 %, p < 0.05)
(Fig. 6b), indicating the higher aquatic toxicity of NaClO-treated
leachates. Polymer type, whether UV/HP- or NaClO-treated, had no
significant effect on cytotoxicity and bacterial toxicity. Given that the
leached DOC concentrations under UV/HP and chlorination were
similar (Fig. S7a), we hypothesized that this conflicting toxicity result
could arise from the different compositions of MPs-derived DOM
generated during two treatments. The observation that the leachates
containing specific organic mixtures exhibited high cytotoxicity and
aquatic toxicity partially supports our hypothesis, although the contri­
butions of individual compounds to the overall toxicity were unable to
be explained in this study.
4. Conclusions
To sum up, we investigated the removal and leaching behavior of
fiber and fragment MPs in different water and wastewater treatment
processes, employing more realistic conditions with actual wastewater
matrices and lower concentrations of MPs. The behavior of MPs in
WWTPs was dependent on both treatment process and intrinsic char­
acteristics of the MPs. Activated sludge process and coagulation
removed MPs from wastewater with efficiencies ranging from 40 % to 94
%, implying the transfer of these MPs to the sludge phase rather than
their treatment. Sand filtration exhibited a high removal efficiency of
over 97 % for MPs, irrespective of their shape or polymer type. Never­
theless, under actual operating conditions for RSF, there was a notable
release of fragments from porous media, which is expected to result in
these fragments predominantly entering the subsequent treatment sys­
tem. In addition, we found that typical AOP and disinfection conditions
did not affect the physicochemical properties of MPs, but led to changes
in their leaching characteristics. The difference in the toxicity and
composition of MP leachates generated during UV-based AOP and
S.-H. Na et al.
chlorination could have implications regarding risk assessments of
weathered MPs, because different tertiary treatment processes may
leach different types of DOM from MPs. Combined, our work expands
our understanding of the fate of MPs in WWTPs and their potential
impacts on aquatic environments and human health. We anticipate that
these findings will provide valuable insights for deriving an effective MP
control strategy to improve the quality of treated water, and will also
boost more attention to the management of MPs-enriched sludge pro­
duced within WWTPs.
CRediT authorship contribution statement
S. H. Na: Investigation, Writing-original draft. M. J. Kim: Concep­
tualization, Investigation, Writing-original draft. J. Kim: Investigation,
Data curation. R. Batool: Investigation. K. Cho: Methodology, Investi­
gation. J. Chung: Validation. S. Lee: Validation, Funding acquisition. E.
J. Kim: Conceptualization, Data curation, Funding acquisition, Super­
vision, Writing-original draft.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data Availability
Data will be made available on request.
Acknowledgements
This work was supported by the National Research Foundation of
Korea (NRF) grant funded by Korea government (2023R1A2C2004069),
’Land/Sea-based input and fate of microplastics in the marine environ­
ment’ of Korea Institute of Marine Science & Technology Promotion
(KIMST) funded by the Ministry of Oceans and Fisheries, Republic of
Korea (no. 20220357), and Korea Institute of Science and Technology
institutional program (2E32442 and 2E32445).
Appendix A. Supporting information
Supplementary data associated with this article can be found in the
online version at doi:10.1016/j.jhazmat.2023.132938.
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