Science of the Total Environment 715 (2020) 136924

Contents lists available at ScienceDirect

Science of the Total Environment

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

Identification and quantification of selected plastics in biosolids by

pressurized liquid extraction combined with double-shot pyrolysis gas
chromatography–mass spectrometry
Elvis D. Okoffo a,⁎, Francisca Ribeiro a,b, Jake W. O'Brien a, Stacey O'Brien a, Benjamin J. Tscharke a,
Michael Gallen a, Saer Samanipour c, Jochen F. Mueller a, Kevin V. Thomas a
a
Queensland Alliance for Environmental Health Sciences (QAEHS), The University of Queensland, 20 Cornwall Street, Woolloongabba, QLD 4102, Australia
b
College of Life and Environmental Sciences, University of Exeter, EX4 4QD Exeter, UK
c
Norwegian Institute for Water Research (NIVA), 0349 Oslo, Norway

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

• Alternative and complementary quanti-

tative analytical method for the analysis
of selected plastics in complex environ-
mental samples.
• Pressurized liquid extraction (PLE) com-
bined with double-shot pyrolysis gas
chromatography-mass spectrometry
(Pyr-GC/MS) allows for the identifica-
tion and quantification of polyethylene,
polystyrene, poly-methyl methacrylate,
polypropylene and polyvinyl chloride.
• The double-shot feature allows for the
effective thermal desorption of poten-
tially interfering co-extracted com-
pounds from complex organic rich
samples.
• Total plastic concentration of between
2.8 and 6.6 mg/g dw (median = 4.1
mg/g dw) in Australian biosolids
• PE was the predominant plastic de-
tected (mean concentration of 2.2 mg/
g dw), contributing to 50 % of the total
of all plastics.

a r t i c l e i n f o a b s t r a c t

Article history: The identification and quantification of selected plastics (polystyrene (PS), polycarbonate (PC), poly-(methyl
Received 21 December 2019 methacrylate) (PMMA), polypropylene (PP), polyethylene terephthalate (PET), polyethylene (PE) and polyvinyl
Received in revised form 23 January 2020 chloride (PVC)) in biosolids (treated sewage sludge) was performed by pressurized liquid extraction (PLE) com-
Accepted 23 January 2020
bined with double-shot pyrolysis gas chromatography–mass spectrometry. Validation of the method yielded re-
Available online 24 January 2020
coveries of between 85 and 128% (mean RSD 11%) at a linear range of between 0.01 and 2 μg. The distribution of
Editor: Damia Barcelo plastics within 25 biosolid samples from a single wastewater treatment plant in Australia was assessed. The mass
concentration of PE, PVC, PP, PS and PMMA was between 0.1 and 4.1 mg/g dry weight (dw) across all samples,

⁎ Corresponding author.
E-mail address: e.okoffo@uq.edu.au (E.D. Okoffo).

https://doi.org/10.1016/j.scitotenv.2020.136924
0048-9697/© 2020 Elsevier B.V. All rights reserved.
2 E.D. Okoffo et al. / Science of the Total Environment 715 (2020) 136924
Keywords: with a total plastic concentration ƩPlastics of between 2.8 and 6.6 mg/g dw (median = 4.1 mg/g dw). PE was the
Plastics predominant plastic detected (mean concentration of 2.2 mg/g dw), contributing to 50% of the total of all plastics.
Pressurized liquid extraction Overall, this study demonstrates that pressurized liquid extraction (PLE) combined with double-shot pyrolysis
Double-shot Pyr-GC/MS
gas chromatography–mass spectrometry can be used to identify and quantify PE, PP, PVC, PS, and PMMA in
Quantification
Sewage sludge
biosolids.
Biosolids
1. Introduction
Awareness that the natural environment is contaminated with small
plastic particles has increased over the past years (Rochman, 2018;
Dümichen et al., 2017; McCormick et al., 2014). Large amounts of plastic
debris accumulate in the environment due to a combination of high pro-
duction, low recycling volumes and the highly stable nature of most
plastic materials (Wang et al., 2017; Ribeiro et al., 2019). Environmental
factors such as sunlight, mechanical stress and an oxidizing atmosphere
(Dümichen et al., 2017), can fragment plastic debris into smaller pieces,
with fragments b 5 mm typically defined as microplastics and frag-
ments b 1 μm termed nanoplastics (Zhou et al., 2019). Plastic particles
have been reported to occur in marine waters (Jambeck et al., 2015;
Ter Halle et al., 2017), freshwaters (Jabeen et al., 2017), terrestrial envi-
ronments (Wang et al., 2017; Huerta Lwanga et al., 2016; Zubris and
Richards, 2005), air (Gasperi et al., 2018; Allen et al., 2019) and living or-
ganisms (Ribeiro et al., 2019; Jabeen et al., 2017; de Sá et al., 2018), with
their potential risks to organisms and human health identified as an
emerging concern (Ziajahromi et al., 2018; Ziajahromi et al., 2017;
Wright and Kelly, 2017).
Biosolid (treated sewage sludge), a product of wastewater treat-
ment, is an important and potentially significant source of plastic to
the terrestrial environment (Nizzetto et al., 2016a; Okoffo et al., 2019;
Mason et al., 2016). They are applied to vast expanses of agricultural
land in order to recycle organic matter, nutrients and to improve soil
quality for cropping (Nizzetto et al., 2016a; Hurley and Nizzetto,
2018). Between 80 and 99% of the plastic particles entering wastewater
treatment plants (WWTPs) have been reported to be retained in bio-
solids (Murphy et al., 2016; Lares et al., 2018; Talvitie et al., 2017;
Mahon et al., 2017). In Australia, Europe and North America, approxi-
mately 50 to 75% of biosolids are recycled via agricultural land applica-
tion (Nizzetto et al., 2016a; Australian Water Association (AWA), 2017)
potentially releasing 2800 to 430,000 tons of plastic annually onto farm-
lands (Nizzetto et al., 2016a; Okoffo et al., 2019; Nizzetto et al., 2016b;
Ng et al., 2018) with unknown consequences for sustainability and
food security (Nizzetto et al., 2016a).
Several recent studies have tried to subjectively quantify the
amount of plastics in biosolids (Okoffo et al., 2019; Raju et al.,
2018; Li et al., 2018; Sun et al., 2019; Prata, 2018; Gatidou et al.,
2019). This has typically been performed by visual inspection and
particle counting, followed by identification using spectroscopic
techniques such as Raman Spectroscopy and Fourier-transform in-
frared (FT-IR) spectroscopy on a subset of particles (Lares et al.,
2018; Mahon et al., 2017; Sun et al., 2019; Gatidou et al., 2019; Hu
et al., 2019; Prata et al., 2019; Gies et al., 2018). While these
approaches provide data on the types of plastic, the number of
particles, size, shape and color, they can't quantify the mass concen-
tration of the plastics in the samples (Eisentraut et al., 2018). In ad-
dition, the above mentioned techniques are size dependent, and in
many cases, not able to detect smaller sized plastics – potentially
underestimating amount of plastics (Okoffo et al., 2019). For exam-
ple, FT-IR and Raman require particle sizes of N20 and N1 μm respec-
tively (Ivleva et al., 2017). For complex matrices, e.g. biosolids, these
techniques require exhaustive and laborious pre-treatment proce-
dures to remove interferences (organic materials) (Okoffo et al.,
2019; Lares et al., 2018; Lares et al., 2019; Mintenig et al., 2017).
© 2020 Elsevier B.V. All rights reserved.
Thermo-analytical techniques, such as pyrolysis-gas chromatogra-
phy coupled with mass spectrometry (Pyr-GC/MS), provide an alterna-
tive for the identification and quantification of plastic – independent of
particle size (Ter Halle et al., 2017; Okoffo et al., 2019; Fischer and
Scholz-Böttcher, 2017; Fischer and Scholz-Böttcher, 2019; Unice et al.,
2013; Hermabessiere et al., 2018). Pyr-GC/MS uses polymer specific de-
composition compounds for identification and facilitates semi-
quantitative to quantitative analyses of plastics in environmental sam-
ples (Fischer and Scholz-Böttcher, 2017; Fischer and Scholz-Böttcher,
2019; Dierkes et al., 2019). Nonetheless, the pre-concentration of plastic
particles in environmental samples to meet the requirement of analysis
by Pyr-GC/MS is not trivial (Zhou et al., 2019). Similar to the use of FT-IR
and Raman, various studies for example have pre-treated environmen-
tal samples by either degrading organic materials using chemical or en-
zymatic digestion and/or by pre-concentrating and separation of plastic
particles prior to Pyr-GC/MS analysis (Ter Halle et al., 2017; Fischer and
Scholz-Böttcher, 2017; Fischer and Scholz-Böttcher, 2019; Dierkes et al.,
2019; Hendrickson et al., 2018). Such clean-up procedures are time-
consuming and may be prone to errors (Zhou et al., 2019).
Sequential pressurized liquid extraction (PLE) has been recently re-
ported as a promising technique for the extraction of selected plastics
(polyethylene (PE), polyvinyl chloride (PVC), polystyrene (PS), poly-
propylene (PP) and polyethylene terephthalate (PET)) from municipal
waste, soil and sediment samples (Dierkes et al., 2019; Fuller and
Gautam, 2016). To reduce the interference from the complex organic
sample matrix, methanol was used at 100 °C as an initial extraction
step and discarded allowing the plastics to be extracted with dichloro-
methane (DCM) at 180 °C (Fuller and Gautam, 2016) or tetrahydrofuran
(THF) at 185 °C (Dierkes et al., 2019). The extract was then either ana-
lyzed gravimetrically on the evaporated extracted residues with plastic
identification confirmed by FT-IR (Fuller and Gautam, 2016) or alterna-
tively by Pyr-GC/MS (Dierkes et al., 2019). While a sequential extraction
clean-up is promising, there is the potential that an initial pre-
extraction step with methanol at 100 °C may remove some plastics
leading to underestimation.
In this paper we present a quantitative method for the extraction
and analysis of seven plastics (PS, PC, PMMA, PP, PET, PE and PVC) in
biosolid samples by combining a single step PLE with a two-stage (dou-
ble-shot) Pyr-GC/MS method that negates the need for a pre-extraction
clean-up step or sample pre-treatment. Here, the first-shot component
of a double-shot Pyr-GC/MS method was used to thermally desorb the
potentially interfering organic compounds/materials co-extracted
from biosolid samples, with plastic identification and quantification
performed in the second-shot. The validated method was applied to a
set of biosolid samples collected from a single treatment train and the
distribution of plastics evaluated.
2. Experimental section
2.1. Reference plastic standards and chemicals
Plastic reference standards: PE (30–530 μm), PS (40–510 μm),
PMMA (30–530 μm), PC (20–520 μm), PET (20–510 μm) and PP
(20–520 μm) were provided by the Norwegian Institute for Water Re-
search (NIVA, Oslo, Norway). PVC powder with size ≤ 50 μm was pur-
chased from Sigma-Aldrich (St. Louis, MO, USA). Each plastic type was
 E.D. Okoffo et al. / Science of the Total Environment 715 (2020) 136924
confirmed by FT-IR spectrometer (PerkinElmer, UATR Two) and Pyr-
GC/MS prior to use (see Supporting information (SI) Fig. S13 and
Table S3). Analytical grade dichloromethane (DCM) was purchased
from Merck (Darmstadt, Germany). Hydromatrix was purchased from
Agilent Technologies (Santa Clara, CA, USA) and pre-extracted with
DCM before use.
2.2. Sample collection
A total of 25 biosolid samples were collected from the solar hall of a
WWTP located in South-East Queensland, Australia. For sampling pur-
poses the solar hall was divided into a grid (9 columns by 30 rows)
with each row representing approximately 1 day of drying. Samples
were collected from approximately every third day of drying (see SI
Fig. S1 for sampling design) using a stainless-steel shovel and collected
into a pre-rinsed (distilled water and acetone) glass jar and stored at
−20 °C until analysis. Ten samples were collected along the days of dry-
ing in column 5 (referred to as T1 to T10) and 15 samples collected ran-
domly across the hall (R1 to R15) (SI Fig. S1). The purpose of this
sampling strategy was to 1) evaluate and understand the natural distri-
bution in plastics content among biosolid samples from a single WWTP;
2) provide a more accurate assessment of the identity and concentra-
tions of plastics present; and 3) provide guidance on the timing and fre-
quency of sample collection.
2.3. Pressurized liquid extraction
Samples were freeze-dried, milled to fine powder with a commercial
grinder for 30 min (Extech equipment Pty. Ltd., Victoria, Australia) and
shaken using an overhead shaker at 140 rpm for 2 h to homogenize. Fol-
lowing this, ~1 g of each sample was extracted by PLE in pre-cleaned
34 ml ASE cells on an ASE 350 system (Dionex, Sunnyvale, CA). Extrac-
tion was performed with DCM at 180 °C and 1500 psi with a heat and
static-time of 5 min using three extraction cycles. Extraction parameters
used in this study are presented in Table 1, with detailed PLE optimiza-
tion and validation procedure presented in SI Table S1. The extracts (in
solvent) were weighed and 80 μl transferred to a pyrolysis cup (Eco-Cup
LF, Frontier Laboratories, Japan) for Pyr-GC/MS analysis. The solvent
was evaporated at room temperature for 30 min prior to analysis. It
should be noted that the evaporation of the solvent was done in a
fume hood to avoid airborne contamination.
2.4. Pyrolysis-GC/MS analysis
Plastic identification and quantification was performed using a
double-shot component of a multi-shot micro-furnace pyrolyzer
(EGA/PY-3030D) equipped with an auto-shot sampler (AS-1020E)
(both Frontier Lab Ltd., Fukushima, Japan) coupled to a GC/MS -
QP2010-Plus (Shimadzu Corporation, Japan). The double-shot Pyr-GC/
MS technique provides the opportunity for selective sample purification
(Quénéa et al., 2006; Terán et al., 2009), as it allows a single sample to be
analyzed twice under different temperature conditions, providing a fast
Table 1
Optimized PLE conditions for the selected plastics.
Parameter Optimized extraction parameters
Extraction solvent Dichloromethane
Extraction temperature (°C) 180
Static time (s) 5
Cycles 3
Rinse volume (%) 80
Purge time (s) 75
System rinse volume (ml) 9 characteristic PE triplets (alkadiene, n-alkene and n-alkane); (2) the
Heating time (min) 9 presence of a homologous series of N5 triplets within C7–C41 of PE stan-
Pressure 1500 psi
3
and reliable procedure to remove organic materials and analyze plastics
in samples (Terán et al., 2009; Derenne and Quénéa, 2015). The first-
shot (thermal desorption), was conducted with a starting temperature
of 100 °C, ramped up to 300 °C at 20 °C min−1, then held at 300 °C for
1 min. The second shot (pyrolysis) was conducted at 650 °C for
0.20 min (12 s). The pyrolyzer interface and GC injection port tempera-
tures were set at 320 and 300 °C, respectively. The samples were
injected with a split of 50:1 on an Ultra Alloy® 5 capillary column
(30 m, 0.25 mm I.D., 0.25 μm film thickness) (Frontier Lab). The GC
oven column temperature program was as follows: held at 40 °C for
2 min, increased to 320 °C at 20 °C min−1, and held for 14 min. Helium
was used as the carrier gas at a 1.0 ml/min with a constant linear veloc-
ity. The ion source temperature was kept at 250 °C with an ionization
voltage of 70 eV. Scan mode was used with a mass range from 40 to
600 m/z (see SI Table S2 for Pyr-GC/MS optimization procedure).
2.5. Plastic indicator compound selection
To identify and quantify single plastics in complex environmental
samples, specific indicator compounds for each plastic are required
(Fischer and Scholz-Böttcher, 2017; Fischer and Scholz-Böttcher,
2019; Hermabessiere et al., 2018; Dierkes et al., 2019; Gomiero et al.,
2019). Plastic specific indicator compounds for PC, PE, PET, PMMA, PP,
PS and PVC were determined by analyzing individual reference stan-
dards and comparing the pyrograms against literature data (Zhou
et al., 2019; Fischer and Scholz-Böttcher, 2017; Fischer and Scholz-
Böttcher, 2019; Hermabessiere et al., 2018; Tsuge et al., 2011), as well
as assessing their specificity against a number of natural materials: -
prawns (chitin), wood, pine needles, humic acid, cellulose (lab filter
paper), fish filet, engine oil, rice and leaves. 1-decene (m/z 83) was cho-
sen as the indicator compound for PE as it was the most representative
pyrolysis product present at a high abundance, although subject to
some minor bias from natural materials (Section 3.3). For PP, 2, 4-
dimethyl-1-heptene (m/z 126) was specific and was selected as an indi-
cator compound. Methyl methacrylate (m/z 100) was specific for PMMA
and was selected as an indicator compound. Although styrene (m/z 104)
was the most abundant indicator compound from the pyrolysis of PS,
natural products, such as chitin, wood (lignin) and fish protein, also re-
leased styrene as pyrolysis product, hence, styrene trimer (m/z 91) was
chosen as the PS specific indicator compound (Zhou et al., 2019; Fischer
and Scholz-Böttcher, 2017) (see Section 3.3). Benzene (m/z 78) was se-
lected as the indicator compound for PVC due the low intensity and sen-
sitivity of the other pyrolysis products. For PC and PET, bisphenol A (m/z
213) and vinyl benzoate (m/z 105) were selected as the indicator com-
pounds, respectively. (See Table 2 for summary and SI Table S3 for fur-
ther detail).
2.6. Quality assurance and quality control
Prior to each PLE extraction batch, the PLE cells were conditioned
with the optimized parameters to remove possible plastic contamina-
tion. Similarly, prior to each Pyr-GC/MS analysis batch, a new pyrolysis
cup blank (instrument blank) and a procedural blank were used to
demonstrate the absence of detectable plastics in the instrument or
method before sample analysis. Procedural blanks were prepared by
extracting hydromatrix with the PLE method and then treating the
clean hydromatrix as a real sample by including in each batch of biosolid
samples to undergo all procedures. No plastic type was identified in the
procedural or instrument blanks (SI Fig. S7). Additionally, all plastics
components of the PLE instrument were sampled directly into pyrolysis
cups and analyzed on the Pyr-GC/MS but no potential contamination
was found. Three validation criteria were used for confirming the pres-
ence of PE in samples: (1) the presence of a homologous series of the
dard and (3) the standard deviation of the peak areas of the individual
4 E.D. Okoffo et al. / Science of the Total Environment 715 (2020) 136924

Table 2
Plastic indicator compounds and ions.

Plastic Pyrolysis product Indicator compounds (m/z) Retention time (min)

Polypropylene 2,4-Dimethyl-1-heptene 70, 83, 126 5.15

Polystyrene 5-Hexene-1,3,5-triyltribenzene (styrene trimer) 91, 117, 194, 312 16.77
Poly-(methyl methacrylate) Methyl methacrylate 69, 100, 89 3.41
Polyethylene terephthalate Vinyl benzoate 105, 77, 148, 51 8.42
Polycarbonate Bisphenol A (BA) 213, 119, 91, 165, 288 15.41
Polyethylene 1-Decene (C10) 83, 97, 111, 140 6.97
Polyvinyl chloride Benzene 78, 74, 52 2.86

Italics and bold values used for calibration and quantification.

C10 triplet is within 2 times the standard deviation of PE standard (n =
5).
3. Results and discussion
3.1. Optimization of experimental parameters
The extraction of seven common plastics (PC, PE, PET, PMMA, PP, PS
and PVC) by PLE was optimized and validated individually via modifica-
tion of the method previously reported by (Fuller and Gautam, 2016) as
outlined in SI Table S1. A single step extraction procedure using DCM at
180 °C and 1500 psi was validated because PC, PMMA and PS were
quantitatively recovered (N93%, n = 5) at 100 °C whereas PE, PET, PP
and PVC required 180 °C to be quantitatively recovered (N90%)
(Table 3, and SI Table S4). Recoveries were measured by dry weight of
the extracts. FT-IR and Pyr-GC/MS analysis of the extracted residues
were similar to that of the original spiking standards, indicating that
no significant chemical changes had occurred during the extraction pro-
cesses (Fuller and Gautam, 2016) (see SI Figs. S13 and S4). Average re-
coveries N 80% were obtained for plastics-spiked biosolid samples
(Table 3, and SI Table S5). The recovery results confirm that PLE extrac-
tion is suitable for the rapid extraction of plastics from biosolids (Fuller
and Gautam, 2016). Reproducibility analysis of the optimized PLE con-
ditions performed on 3 separate occasions was between 1.4 and 10.6%
(RSD, %) (SI Table S6), confirming that the PLE extraction conditions
are accurate and reproducible over time.
Given that PE, PET and PP are poorly soluble in DCM at room temper-
ature, a dissolution analysis of PLE extracted PE, PET and PP was per-
formed over 3 h post extraction. PE, PET and PP remained in solution
over this time (RSD of b20% for each) allowing for sufficient time to al-
iquot into the pyrolysis cup for analysis (see Fig. 1 for dissolution
analysis of PLE extracted PS, PVC, PP, PMMA, PC, PET and PE at multiple
time points post extraction).
Double-shot Pyr-GC/MS was optimized and validated for the analy-
sis of PC, PE, PET, PMMA, PP, PS and PVC. The first-shot was optimized to
thermally desorb co-extracted potential interferences (organic mate-
rials) that may potentially interfere with the pyrogram without
degrading the target plastics. This was conducted through progressive
heating of the sample extract (100–300 °C, 100–320 °C and
100–340 °C) and confirming that no plastic was measured. Similarly,
the second-shot (pyrolysis) temperature was optimized to pyrolyze
plastics – PC, PMMA and PS (550, 600, 650 and 700 °C) and assessed
by comparing peak areas of their indicator compounds. The decomposi-
tion products of all plastic standards were stable up to the pyrolysis
temperature of 650 °C (e.g. SI Fig. S2). None of the decomposition prod-
ucts of plastics-spiked in biosolids were observed in the ion chromato-
gram of the thermally desorbed organic fraction. An optimum
temperature of 100–300 °C was chosen for the thermal desorption
step (first-shot) as it had no measurable effect on the accuracy of plastic
identification and quantification. The absolute peak areas of all the py-
rolysis indicator compounds of PC, PMMA and PS were significantly im-
pacted by changes to the pyrolysis furnace temperatures. A rise in
response was observed when the pyrolysis temperature was increased
from 550 to 650 °C but declined at 700 °C (SI Fig. S3). The optimum tem-
perature for pyrolysis analysis was found at 650 °C since it revealed
higher peak areas for all the indicator compounds.
3.2. Method performance and validation
An external calibration curve was prepared by weighing plastic stan-
dards (from 0.5 to 100 μg), mixing with pre-washed hydromatrix
(milled to fine powder), PLE extracted and aliquoted into pyrolysis

Table 3
Summary of method performance.

Plastic Recovery RSD Recovery RSD Indicator compound Linear range (μg LOQ Calibration Linearity Intradayc Inter dayc
type (n = 5)a (%) (%) (n = 5)b (%) (%) on column)c (mg/g)c functionc (r2)c (n = 5) (n = 5)
RSD (%) RSD (%)

PE 91 ± 8 8.4 83 ± 3 3.6 1-Decene 0.02–2 0.03 y = 49,370 0.939 12.0 6.1

x − 1242.1
PMMA 98 ± 3 3.4 91 ± 2 1.9 Methyl methacrylate 0.06–2 0.09 y = 20,000,000 0.956 5.1 4.1
x − 2,000,000
PS 98 ± 3 13.7 93 ± 3 2.9 5-Hexene-1,3,5-triyltribenzene 0.01–2 0.01 y = 2,000,000 0.995 4.6 3.6
(PS trimer) x − 30,212
PET 93 ± 9 9.4 85 ± 5 6.0 Vinyl benzoate 0.02–2 0.03 y = 152,301 0.961 7.0 4.4
x − 9309.1
PC 99 ± 8 8.3 95 ± 2 2.3 Bisphenol A 0.02–2 0.03 y = 59,126 0.961 8.1 5.2
x − 192,949
PP 94 ± 5 5.6 85 ± 4 4.5 2,4-Dimethyl-1-heptene 0.02–2 0.03 y = 26,265 0.997 9.8 3.1
x − 1241.8
PVC 87 ± 8 8.6 84 ± 3 3.1 Benzene 0.02–2 0.03 y = 17,360 0.960 7.4 4.8
x − 82,470

n = indicates number of analyzed samples. Polystyrene (PS), polycarbonate (PC), poly-(methyl methacrylate) (PMMA), polypropylene (PP), polyethylene terephthalate (PET), polyeth-
ylene (PE) and polyvinyl chloride (PVC).
a
Recovery results from plastic spiked standards extracted on PLE.
b
Recovery results from plastic spiked biosolid extracted on PLE.
c
Results are based on the selected indicator compounds for the plastic as presented in Table 2 and SI Table S3.
 E.D. Okoffo et al. / Science of the Total Environment 715 (2020) 136924 5

PE PET PC
400000 300000 240000

Peak areas

Peak areas
Peak areas

350000 250000 220000

300000 200000 200000

250000 150000 180000

5
25

75

5
25

75
5

5
25

75

2
0.

1.

2.

2
0

0.

1.

2.
0.

1.

2.

0.

0.

0.

0.
0.

0.

Time (h) Time (h) Time (h)

PS PP
PMMA
1750000 1600000
3600000
1700000
3400000
Peak areas

1400000

Peak areas
Peak areas

1650000
3200000
1200000
3000000 1600000

1550000 1000000
2800000

2600000 1500000 800000

5

5
25

75

25

75
0

2
5

5
25

75

0.

1.

2.

0.

1.

2.
0

2
0.

1.

2.

0.

0.

0.

0.
0.

0.

Time (h) Time (h) Time (h)

PVC
950000

900000
Peak areas

850000

800000

750000

700000
5

5
25

75
0

2
0.

1.

2.
0.

0.

Time (h)

Fig. 1. Dissolution analysis of PLE extracted PS, PVC, PP, PMMA, PC, PET and PE at multiple time points post extraction (0, 0.25, 0.5, 0.75, 1, 1.5, 2, and 2.5 h). Polystyrene (PS), polycarbonate
(PC), poly-(methyl methacrylate) (PMMA), polypropylene (PP), polyethylene terephthalate (PET), polyethylene (PE) and polyvinyl chloride (PVC).

cups. With a split of 50:1 the calibration range of each plastic was from
0.01 to 2 μg on column having R2 ≥ 0.93 (Table 3 and SI Fig. S11). Plastic
standards were measured individually and in mixtures. The limit of
quantification (LOQ) for each plastic was defined as the lowest detect-
able standard (lowest concentration) of the calibration curve and also
in pooled biosolid which produced a signal 10 times the baseline noise
(S/N 10) and where the RSD of 7 replicate injections was b20%
(Table 3). Intra- and inter day variation was calculated as the RSD % of
five repeated analyses of a plastic (5 μg) on the same day and over
5 days respectively, with precision between 3.1 and 12.0% (Table 3).
Three recovery experiments containing all seven reference plastics at
known concentrations (20 to 100 μg), were included in this study and
underwent the same treatment as the biosolid samples (see SI
Table S10 for detail spiking procedure). Acceptable mean recoveries
were between 84% and 109% (n = 3) for all plastics (SI Table S7). Matrix
interferences were investigated by spiking four of the analyzed biosolid
samples with concentrations of plastics (20 to 100 μg) (see SI Table S10
for detail spiking procedure). The mean recoveries of the spiked plastics
ranged from 85% to 128% (n = 4) (SI Table S8) indicating that the results
were not influenced by matrix interferences. Method performance was
evaluated by analyzing extracted PS standard and PS spiked biosolid,
with an acceptance criterion of b20% drift from the spiked concentra-
tion. Method reproducibility was assessed by a laboratory duplicate
analysis of selected biosolid samples, with comparable levels observed
for all samples (Fig. 3).
3.3. Potential matrix interferences
Natural materials are a potential source of interfering indicator com-
pounds during pyrolysis (Fischer and Scholz-Böttcher, 2017; Dierkes
6 E.D. Okoffo et al. / Science of the Total Environment 715 (2020) 136924

Table 4
Potential interferences of organic materials with plastics selected indicator compounds.

Matrix Substance Concentration of indicator compound (mg/g)a

compound class
PE PP PS (trimer) PVC PET (vinyl PC PMMA (methyl
(1-decene (2,4-dimethyl-1-heptene (5-hexene-1,3,5-triyltribenzene (benzene benzoate (bisphenol methacrylate
(C10) (m/z (m/z 126)) (m/z 91)) (m/z 78)) (m/z A (m/z
83)) 105)) (m/z 213)) 100))

Fish filet Proteins, fat 0.05 b0.03 b0.01 b0.03 b0.03 b0.03 b0.09
Fir needle Tree gum, terpene b0.03 b0.03 b0.01 b0.03 b0.03 b0.03 b0.09
Humic acids Organic matter 0.03 b0.03 b0.01 b0.03 b0.03 b0.03 b0.09
Prawn Chitin, proteins 0.04 b0.03 b0.01 b0.03 b0.03 b0.03 b0.09
Wood Lignin 0.04 b0.03 b0.01 b0.03 b0.03 b0.03 b0.09
Engine oil Hydrocarbons 0.04 b0.03 b0.01 b0.03 b0.03 b0.03 b0.09
Leaf Cellulose, organic matter b0.03 b0.03 b0.01 b0.03 b0.03 b0.03 b0.09
Filter paper Cellulose b0.03 b0.03 b0.01 b0.03 b0.03 b0.03 b0.09
Rice Carbohydrate b0.03 b0.03 b0.01 b0.03 b0.03 b0.03 b0.09
a
Matrix-induced bias in mg of plastic by 1 g of matrix. Polystyrene (PS), polycarbonate (PC), poly-(methyl methacrylate) (PMMA), polypropylene (PP), polyethylene terephthalate
(PET), polyethylene (PE) and polyvinyl chloride (PVC). LOQs in mg/g for PS (0.01), PMMA (0.09), PE (0.03), PET (0.03), PC (0.03), PVC (0.03) and PP (0.03).

et al., 2019). The potential for their formation was evaluated by analyz- East Queensland, Australia. The pyrograms of the analyzed biosolid
ing a number of organic materials and biogenic polymers such as chitin samples featured specific compounds clearly related to the presence of
isolated from prawn, wood (lignin), pine needles, humic acid (organic PE, PP, PVC, PS and PMMA (see SI Fig. S6 and Table S9 for example).
matter), cellulose (lab filter paper), fish filet (proteins, fat), engine oil
(hydrocarbons), rice (carbohydrate) and leaves (cellulose, organic mat-
ter) as previously reported by (Fischer and Scholz-Böttcher, 2017; Table 5
Dierkes et al., 2019). None of the natural materials produced interfering Concentrations of PE, PP, PVC, PS and PMMA (mg/g dw) measured in biosolid samples.
indicator compounds for PP, PET, PC, PMMA and PS when using thermal
Sample ID PE PP PVC PS PMMA Ʃplastic
desorption (first-shot of the double-shot method) as a clean-up step
(Table 4). Chitin, wood and fish protein released styrene (PS monomer) Samples taken in (T1–T10) along
column 5 (10 samples)
during the pyrolysis process as previously reported (Zhou et al., 2019;
T1.1 1.9 0.7 0.8 bLOQ 0.4 3.8
Fischer and Scholz-Böttcher, 2017; Dierkes et al., 2019), hence, PS tri- T1.2 1.7 0.7 0.8 bLOQ 0.4 3.6
mer is therefore used as the indicator compound for PS. Phenylalanine T2.1 4.0 0.8 0.9 0.5 0.5 6.6
is a known precursor for styrene formation during pyrolysis with no T2.2 4.0 0.7 0.8 0.4 0.5 6.4
T3 1.6 0.4 0.8 0.6 bLOQ 3.4
PS trimer formed (Fischer and Scholz-Böttcher, 2017; Dierkes et al.,
T4 4.0 0.5 1.0 0.3 0.6 6.5
2019). T5.1 1.3 0.6 0.9 bLOQ 0.6 3.4
The formation of various indicator compound interferences for PE T5.2 1.1 0.6 0.9 bLOQ 0.6 3.2
has previously been reported following the pyrolysis of a range of natu- T6 1.0 0.4 0.7 0.7 bLOQ 2.8
ral products (Dierkes et al., 2019). Commonly, biogenic materials such T7 2.8 0.8 0.9 0.5 0.6 5.6
T8 3.0 0.3 0.7 0.9 bLOQ 4.8
as natural fats (e.g. fish protein) and waxes that are rich in long alkyl
T9 1.0 0.6 1.2 0.2 0.9 3.9
chains have been reported to produce n-alkanes and n-alkenes during T10 1.3 0.5 1.2 0.3 0.5 3.8
pyrolysis (Fischer and Scholz-Böttcher, 2017; Dierkes et al., 2019; Samples taken randomly (R1–R15)
Scholz-Böttcher et al., 2013). Most of the materials tested produced across the solar hall (15 samples)
traces of 1-decene, however these background interferences were typi- R1.1 4.0 0.8 0.9 bLOQ bLOQ 5.8
R1.2 3.9 0.9 0.9 bLOQ bLOQ 5.7
cally at or below the LOQ (0.03 mg/g) following using thermal desorp- R2 2.1 0.9 0.7 0.4 bLOQ 4.1
tion as a clean-up step (Table 4). 1-Decene was chosen as an indicator R3 1.9 0.4 0.8 0.4 bLOQ 3.6
compound in this study due to its higher sensitivity and lower LOQ com- R4 1.0 0.7 0.7 0.1 0.4 3.0
pared with other alkanes and alkadiene pyrolysates of PE (Dierkes et al., R5 1.9 1.5 0.8 0.7 bLOQ 4.8
R6 2.6 0.6 0.8 0.8 0.4 5.3
2019). A check of all samples against the three PE validation criteria
R7 4.1 0.3 0.9 0.3 bLOQ 5.6
outlined in Section 2.6 were found to conform, with chain lengths in R8 2.2 0.5 0.9 0.6 bLOQ 4.2
the range of 7–36 carbon atoms. This indicates that the measured PE R9 1.1 0.4 0.9 0.4 bLOQ 2.9
concentration is free of positive or negative bias from matrix effects R10 1.6 0.2 1.0 0.2 bLOQ 3.0
and interfering environmental sources of the matrix compounds. R11 0.7 0.5 0.8 0.4 0.5 2.9
R12.1 3.8 1.4 0.9 bLOQ bLOQ 6.1
Humic acid (organic matter), rice (carbohydrate), fish filet (proteins, R12.2 3.8 1.2 0.9 bLOQ bLOQ 5.9
fat), engine oil (hydrocarbons) and wood (lignin) caused an increased R13 1.7 1.3 0.8 0.3 0.4 4.6
background interference for the analysis of PVC, however these interfer- R14 1.7 0.9 0.8 0.7 bLOQ 4.1
ences were below the LOQ (0.03 mg/g) following the thermal desorp- R15.1 2.7 0.4 1.1 bLOQ 0.6 4.8
R15.2 2.9 0.4 0.9 bLOQ 0.5 4.7
tion clean-up step (Table 4). We therefore accept that the
Mean 2.2 0.7 0.9 0.5 0.5 4.4
quantification of PE and PVC in the biosolid samples are potentially sub- SD 1.1 0.3 0.1 0.2 0.2 1.1
ject to a minor source of positive bias, however these are typically at or Median 1.9 0.6 0.9 0.4 0.5 4.1
below LOQs (Table 4). Sum of plastics (% contribution to total 55 16 22 9 (9) 6 (6) 109
sum of plastics) (50) (15) (20) (100)

3.4. Case study - distribution of plastic concentration in a municipal biosolid
samples
The optimized and validated method was applied to 25 biosolid
samples collected from a WWTP (a single treatment train) in South-
T1.1 and T1.2; T2.1 and T2.2; T5.1 and T5.2; R1.1 and R1.2; R12.1 and R12.2; R15.1 and 15.2
are duplicate analysis, bLOQ: below limit of quantification, LOQs in mg/g for PS (0.01),
PMMA (0.09), PE (0.03), PET (0.03), PC (0.03), PVC (0.03) and PP (0.03), T1–T10; samples
taken along column 5, R1–R15; samples taken randomly across the solar hall (15 samples).
Polystyrene (PS), poly-(methyl methacrylate) (PMMA), polypropylene (PP), polyethylene
(PE) and polyvinyl chloride (PVC).
 E.D. Okoffo et al. / Science of the Total Environment 715 (2020) 136924 7

All samples contained PE, PP and PVC, while PS was found in 80% and
PMMA in 50% of samples. Concentrations of measured plastics ranged
from 0.1 to 4.1 mg/g dw and were consistent between all samples
(Table 5, Fig. 2). PE ranged from 0.7 to 4.1 mg/g dry weight (dw) (me-
dian = 1.9 mg/g dw), PP from 0.2 to 1.5 mg/g dw (median = 0.6 mg/
g dw), PVC from 0.7 to 1.2 mg/g dw (median = 0.9 mg/g dw), PS
from 0.1 to 0.9 mg/g dw (median = 0.4 mg/g dw), and PMMA from
0.4 to 0.9 mg/g dw (median 0.5 mg/g dw) (Table 5, Fig. 2). The average
concentrations of total plastics (ƩPlastics) in the biosolids ranged from
2.8 mg/g dw to 6.6 mg/g dw with a median value of 4.1 mg/g dw
(Fig. 2, Table 5).
PE was the predominant plastic detected in biosolids (mean concen-
tration of 2.2 mg/g dw), contributing to 50% of the total sum of all plas-
tics (Table 5, SI Fig. S12). The total individual sum concentrations of PE,
PP, PVC, PS and PMMA in the biosolid samples ranged from 6 to 55 mg/g
dw (significantly different from each other (p b 0.05, ANOVA)) with a
total sum of 109 mg/g dw (Table 5). Similarly, the total individual sum
of plastics observed for the randomly collected samples (PE = 33 mg/
g dw, PP = 11 mg/g dw, PVC = 13 mg/g dw, PS = 5 mg/g dw and
PMMA = 2 mg/g dw) were slightly higher than the samples taken
along the days of drying (PE = 22 mg/g dw, PVC = 9 mg/g dw, PP =
6 mg/g, PS = 4 mg/g dw, and PMMA = 4 mg/g dw) except for PMMA.
The variance in plastic concentration, both for the sum of all ana-
lyzed plastics and for the individual plastics, between all samples was
relatively low, hence, the distribution throughout the biosolids treat-
ment train can be considered relatively homogenous. To test for repro-
ducibility and sensitivity of the method, duplicate analysis (i.e., splitting
6 samples into two) was as assessed. Analytical reproducibility was
found as RSD = 20% across all detected plastics (Fig. 3). As such analyt-
ical variability was only a very minor source of uncertainty.
4. Conclusions
The results of this work demonstrate that single step PLE coupled
with double-shot Pyr-GC/MS is a suitable method for the rapid and ef-
fective identification and quantification of PE, PP, PVC, PMMA and PS
in biosolid samples and may be suitable for other plastic types and envi-
ronmental matrices. The unique use of the double-shot feature for the
effective thermal desorption of potentially interfering co-extracted
compounds from the biosolid samples provides an improved alternative
for the identification and quantification of plastics in complex organic
rich samples. This reduces processing and labor times needed to pre-
treat samples. The sensitivity of the method proves the advantage of
using Pyr-GC/MS to measure the mass load concentrations of specific
plastics in samples which provides a basis for the uniform reporting of
results as compared to the use of conventional FT-IR and Raman. In par-
ticular, the results are obtained as mass concentration which is a more
standardized and reliable way for comparison of data than particles

Fig. 2. Concentration (mg/g dw) distribution of 5 identified plastics (PE, PMMA, PP, PS, PVC and total plastics) in the 25 biosolid samples collected along the approximate days of drying and
across the solar hall (columns 1–9). NB: sampling along days (Rows) and along column 5 represents samples T1 to T10 (10 samples); sampling along days and across the columns (1 to
9) of the solar hall represents samples taken randomly (R1 to R15) across the hall (15 samples); blank or empty columns/were locations where sample were not collected. Polystyrene
(PS), poly-(methyl methacrylate) (PMMA), polypropylene (PP), polyethylene (PE) and polyvinyl chloride (PVC). LOQs in mg/g for PS (0.01), PMMA (0.09), PE (0.03), PET (0.03), PC
(0.03), PVC (0.03) and PP (0.03).
8 E.D. Okoffo et al. / Science of the Total Environment 715 (2020) 136924

PMMA
PS
PVC
Concentration (mg/g dw)
8
PP
PE

R10
R11
R12.1
R12.2
R13
R14
R15.1
R15.2
T1.1
T1.2
T2.1
T2.2
T3
T4
T5.1
T5.2
T6
T7
T8
T9
T10
R1.1
R1.2
R2
R3
R4
R5
R6
R7
R8
R9
Sampling locations
Fig. 3. Composition (mg/g dw) of detected selected plastics in the 25 biosolid samples collected across the solar hall. T1–T10; samples taken along column 5 R1–R15; samples taken
randomly across the solar hall (15 samples), T1.1 and T1.2; T2.1 and T2.2; T5.1 and T5.2; R1.1 and R1.2; R12.1 and R12.2; R15.1 and 15.2 are duplicate analyses. Polystyrene (PS), poly-
(methyl methacrylate) (PMMA), polypropylene (PP), polyethylene (PE) and polyvinyl chloride (PVC).

number which can be difficult when comparing between locations double-shot pyrograms of extracted sewage sludge; recovery and
(Fischer and Scholz-Böttcher, 2019; Fuller and Gautam, 2016). It should spike experiments on PLE combined with Pyr-GC/MS; and typical char-
however be noted that the mass concentrations provided by this acteristics of real biosolid sample on a double-shot Pyr-GC/MS. Supple-
method are at the same time highly complementary to conventional mentary data to this article can be found online at https://doi.org/10.
FT-IR and Raman analysis of particle counts that provides information 1016/j.scitotenv.2020.136924.
on particle size, shape, and color (Fischer and Scholz-Böttcher, 2019;
Fuller and Gautam, 2016). References

Allen, S., Allen, D., Phoenix, V.R., Le Roux, G., Durántez Jiménez, P., Simonneau, A., et al.,
Declaration of competing interest 2019. Atmospheric transport and deposition of microplastics in a remote mountain
catchment. Nature Geoscience 12 (5), 339–344.
The authors declare that they have no known competing financial Australian Water Association (AWA), 2017. Australian biosolids statistics. Australia &
New Zealand Biosolids Partnership, Australia [cited 10/03/2019]. Available from.
interests or personal relationships that could have appeared to influ- https://www.biosolids.com.au/guidelines/australian-biosolids-statistics/.
ence the work reported in this paper. Derenne, S., Quénéa, K., 2015. Analytical pyrolysis as a tool to probe soil organic matter.
Journal of Analytical and Applied Pyrolysis 111, 108–120.
Dierkes, G., Lauschke, T., Becher, S., Schumacher, H., Földi, C., Ternes, T., 2019. Quantifica-
Acknowledgements tion of microplastics in environmental samples via pressurized liquid extraction and
pyrolysis-gas chromatography. Analytical and Bioanalytical Chemistry 411,
Sincere thanks to Chang He, Sicco H. Brandsma and Xianyu Wang for 6959–6968. https://doi.org/10.1007/s00216-019-02066-9.
Dümichen, E., Eisentraut, P., Bannick, C.G., Barthel, A.-K., Senz, R., Braun, U., 2017. Fast
the technical assistance, Robert Gray for constructive comments, Jack
identification of microplastics in complex environmental samples by a thermal deg-
Thompson for helpful discussions, the Norwegian Institute for Water radation method. Chemosphere 174, 572–584.
Research (NIVA, Oslo, Norway) for kindly providing the plastic refer- Eisentraut, P., Dümichen, E., Ruhl, A.S., Jekel, M., Albrecht, M., Gehde, M., et al., 2018. Two
ence standards, and the operators of the WWTP who provided informa- birds with one stone—fast and simultaneous analysis of microplastics: microparticles
derived from thermoplastics and tire wear. Environmental Science & Technology Let-
tion and allowed us to collect samples. EDO is supported by a Research ters 5 (10), 608–613.
Training Scholarship awarded by the Queensland Alliance for Environ- Fischer, M., Scholz-Böttcher, B.M., 2017. Simultaneous trace identification and quantifica-
mental Health Sciences (QAEHS) and The University of Queensland. FR tion of common types of microplastics in environmental samples by pyrolysis-gas
chromatography–mass spectrometry. Environmental Science & Technology 51 (9),
was supported through a QUEX PhD scholarship - a joint initiative of 5052–5060.
The University of Queensland and the University of Exeter. The Queens- Fischer, M., Scholz-Böttcher, B.M., 2019. Microplastics analysis in environmental samples
land Alliance for Environmental Health Sciences, The University of – recent pyrolysis-gas chromatography-mass spectrometry method improvements
to increase the reliability of mass related data. Analytical Methods 11, 2489–2497.
Queensland, gratefully acknowledges the financial support of the Fuller, S., Gautam, A., 2016. A procedure for measuring microplastics using pressurized
Queensland Department of Health. fluid extraction. Environmental Science & Technology 50 (11), 5774–5780.
Gasperi, J., Wright, S.L., Dris, R., Collard, F., Mandin, C., Guerrouache, M., et al., 2018.
Microplastics in air: are we breathing it in? Current Opinion in Environmental Sci-
Appendix A. Supplementary data ence & Health 1, 1–5.
Gatidou, G., Arvaniti, O.S., Stasinakis, A.S., 2019. Review on the occurrence and fate of
Additional information on sampling design; optimization procedure microplastics in Sewage Treatment Plants. Journal of Hazardous Materials 367,
504–512.
of PLE; optimization procedure of Double-Shot Pyrolysis method; recov-
Gies, E.A., LeNoble, J.L., Noël, M., Etemadifar, A., Bishay, F., Hall, E.R., et al., 2018. Retention
ery and spike experiments on PLE; thermal desorption and pyrograms of microplastics in a major secondary wastewater treatment plant in Vancouver,
of PS, PE, PP PET, PMMA and PC at 100–300 °C and 650 °C respectively; Canada. Marine Pollution Bulletin 133, 553–561.
influence of Pyr-GC/MS temperature on peak response of PMMA, PS, Gomiero, A., Øysæd, K.B., Agustsson, T., van Hoytema, N., van Thiel, T., Grati, F., 2019. First
record of characterization, concentration and distribution of microplastics in coastal
and PC; comparison of pyrograms of extracted PP and PE at 180 °C sediments of an urban fjord in south west Norway using a thermal degradation
and the original spiked material; comparison of a single shot and a method. Chemosphere 227, 705–714.
 E.D. Okoffo et al. / Science of the Total Environment 715 (2020) 136924
Hendrickson, E., Minor, E.C., Schreiner, K., 2018. Microplastic abundance and composition
in western Lake Superior as determined via microscopy, Pyr-GC/MS, and FTIR. Envi-
ronmental Science & Technology 52 (4), 1787–1796.
Hermabessiere, L., Himber, C., Boricaud, B., Kazour, M., Amara, R., Cassone, A.-L., et al.,
2018. Optimization, performance, and application of a pyrolysis-GC/MS method for
the identification of microplastics. Analytical and Bioanalytical Chemistry 410 (25),
6663–6676.
Hu, Y., Gong, M., Wang, J., Bassi, A., 2019. Current research trends on microplastic pollu-
tion from wastewater systems: a critical review. Reviews in Environmental Science
and Bio/Technology 18 (2), 207–230.
Huerta Lwanga, E., Gertsen, H., Gooren, H., Peters, P., Salánki, T., van der Ploeg, M., et al.,
2016. Microplastics in the terrestrial ecosystem: implications for Lumbricus terrestris
(Oligochaeta, Lumbricidae). Environmental Science & Technology 50 (5), 2685–2691.
Hurley, R.R., Nizzetto, L., 2018. Fate and occurrence of micro(nano)plastics in soils:
knowledge gaps and possible risks. Current Opinion in Environmental Science &
Health 1, 6–11.
Ivleva, N.P., Wiesheu, A.C., Niessner, R., 2017. Microplastic in aquatic ecosystems.
Angewandte Chemie International Edition 56 (7), 1720–1739.
Jabeen, K., Su, L., Li, J., Yang, D., Tong, C., Mu, J., et al., 2017. Microplastics and mesoplastics
in fish from coastal and fresh waters of China. Environmental Pollution 221, 141–149.
Jambeck, J.R., Geyer, R., Wilcox, C., Siegler, T.R., Perryman, M., Andrady, A., et al., 2015.
Plastic waste inputs from land into the ocean. Science 347 (6223), 768–771.
Lares, M., Ncibi, M.C., Sillanpää, M., Sillanpää, M., 2018. Occurrence, identification and re-
moval of microplastic particles and fibers in conventional activated sludge process
and advanced MBR technology. Water Research 133, 236–246.
Lares, M., Ncibi, M.C., Sillanpää, M., Sillanpää, M., 2019. Intercomparison study on com-
monly used methods to determine microplastics in wastewater and sludge samples.
Environmental Science and Pollution Research 26 (12), 12109–12122.
Li, X., Chen, L., Mei, Q., Dong, B., Dai, X., Ding, G., et al., 2018. Microplastics in sewage
sludge from the wastewater treatment plants in China. Water Research 142, 75–85.
Mahon, A.M., O’Connell, B., Healy, M.G., O’Connor, I., Officer, R., Nash, R., et al., 2017.
Microplastics in sewage sludge: effects of treatment. Environmental Science & Tech-
nology 51 (2), 810–818.
Mason, S.A., Garneau, D., Sutton, R., Chu, Y., Ehmann, K., Barnes, J., et al., 2016. Microplastic
pollution is widely detected in US municipal wastewater treatment plant effluent.
Environmental Pollution 218, 1045–1054.
McCormick, A., Hoellein, T.J., Mason, S.A., Schluep, J., Kelly, J.J., 2014. Microplastic is an
abundant and distinct microbial habitat in an urban river. Environmental Science &
Technology 48 (20), 11863–11871.
Mintenig, S.M., Int-Veen, I., Löder, M.G.J., Primpke, S., Gerdts, G., 2017. Identification of
microplastic in effluents of waste water treatment plants using focal plane array-
based micro-Fourier-transform infrared imaging. Water Research 108, 365–372.
Murphy, F., Ewins, C., Carbonnier, F., Quinn, B., 2016. Wastewater treatment works
(WwTW) as a source of microplastics in the aquatic environment. Environmental Sci-
ence & Technology 50 (11), 5800–5808.
Ng, E.-L., Huerta Lwanga, E., Eldridge, S.M., Johnston, P., Hu, H.-W., Geissen, V., et al., 2018.
An overview of microplastic and nanoplastic pollution in agroecosystems. Science of
The Total Environment 627, 1377–1388.
Nizzetto, L., Futter, M., Langaas, S., 2016a. Are agricultural soils dumps for microplastics of
urban origin? Environmental Science & Technology 50 (20), 10777–10779.
Nizzetto, L., Langaas, S., Futter, M., 2016b. Pollution: do microplastics spill on to farm
soils? Nature 537, 488.
Okoffo, E.D., O’Brien, S., O’Brien, J.W., Tscharke, B.J., Thomas, K.V., 2019. Wastewater treat-
ment plants as a source of plastics in the environment: a review of occurrence,
methods for identification, quantification and fate. Environmental Science: Water Re-
search & Technology 5 (11), 1908–1931.
Prata, J.C., 2018. Microplastics in wastewater: state of the knowledge on sources, fate and
solutions. Marine Pollution Bulletin 129 (1), 262–265.
9
Prata, J.C., da Costa, J.P., Duarte, A.C., Rocha-Santos, T., 2019. Methods for sampling and de-
tection of microplastics in water and sediment: a critical review. TrAC Trends in An-
alytical Chemistry 110, 150–159.
Quénéa, K., Derenne, S., González-Vila, F.J., González-Pérez, J.A., Mariotti, A., Largeau, C.,
2006. Double-shot pyrolysis of the non-hydrolysable organic fraction isolated from
a sandy forest soil (Landes de Gascogne, South-West France): comparison with clas-
sical Curie point pyrolysis. Journal of Analytical and Applied Pyrolysis 76 (1),
271–279.
Raju, S., Carbery, M., Kuttykattil, A., Senathirajah, K., Subashchandrabose, S.R., Evans, G., et
al., 2018. Transport and fate of microplastics in wastewater treatment plants: impli-
cations to environmental health. Reviews in Environmental Science and Bio/Technol-
ogy 17 (4), 637–653.
Ribeiro, F., O’Brien, J.W., Galloway, T., Thomas, K.V., 2019. Accumulation and fate of nano-
and micro-plastics and associated contaminants in organisms. TrAC Trends in Analyt-
ical Chemistry 111, 139–147.
Rochman, C.M., 2018. Microplastics research—from sink to source. Science 360 (6384),
28–29.
de Sá, L.C., Oliveira, M., Ribeiro, F., Rocha, T.L., Futter, M.N., 2018. Studies of the effects of
microplastics on aquatic organisms: what do we know and where should we focus
our efforts in the future? Science of The Total Environment 645, 1029–1039.
Scholz-Böttcher, B.M., Nissenbaum, A., Rullkötter, J., 2013. An 18th century medication
“Mumia vera aegyptica” – fake or authentic? Organic Geochemistry 65, 1–18.
Sun, J., Dai, X., Wang, Q., van Loosdrecht, M.C.M., Ni, B.-J., 2019. Microplastics in wastewa-
ter treatment plants: detection, occurrence and removal. Water Research 152, 21–37.
Talvitie, J., Mikola, A., Setälä, O., Heinonen, M., Koistinen, A., 2017. How well is microlitter
purified from wastewater? – a detailed study on the stepwise removal of microlitter
in a tertiary level wastewater treatment plant. Water Research 109, 164–172.
Ter Halle, A., Jeanneau, L., Martignac, M., Jardé, E., Pedrono, B., Brach, L., et al., 2017.
Nanoplastic in the North Atlantic Subtropical Gyre. Environmental Science & Technol-
ogy 51 (23), 13689–13697.
Terán, A., Gonzalez-Vila, F.J., Gonzalez-Perez, J.A., 2009. Detection of organic contamina-
tion in sediments by double-shoot pyrolysis–GC/MS. Environmental Chemistry Let-
ters 7 (4), 301–308.
Tsuge, S., Ohtani, H., Watanabe, C., 2011. Pyrolysis-GC/MS Data Book of Synthetic Poly-
mers: Pyrograms, Thermograms and MS of Pyrolyzates. Elsevier.
Unice, K.M., Kreider, M.L., Panko, J.M., 2013. Comparison of tire and road wear particle
concentrations in sediment for watersheds in France, Japan, and the United States
by quantitative pyrolysis GC/MS analysis. Environmental Science & Technology 47
(15), 8138–8147.
Wang, L., Zhang, J., Hou, S., Sun, H., 2017. A simple method for quantifying polycarbonate
and polyethylene terephthalate microplastics in environmental samples by liquid
chromatography–tandem mass spectrometry. Environmental Science & Technology
Letters 4 (12), 530–534.
Wright, S.L., Kelly, F.J., 2017. Plastic and human health: a micro issue? Environmental Sci-
ence & Technology 51 (12), 6634–6647.
Zhou, X., Hao, L., Wang, H., Li, Y., Liu, J., 2019. Cloud-point extraction combined with ther-
mal degradation for nanoplastic analysis using pyrolysis gas chromatography–mass
spectrometry. Analytical Chemistry 91 (3), 1785–1790.
Ziajahromi, S., Kumar, A., Neale, P.A., Leusch, F.D.L., 2017. Impact of microplastic beads
and fibers on waterflea (Ceriodaphnia dubia) survival, growth, and reproduction: im-
plications of single and mixture exposures. Environmental Science & Technology 51
(22), 13397–13406.
Ziajahromi, S., Kumar, A., Neale, P.A., Leusch, F.D.L., 2018. Environmentally relevant con-
centrations of polyethylene microplastics negatively impact the survival, growth
and emergence of sediment-dwelling invertebrates. Environmental Pollution 236,
425–431.
Zubris, K.A.V., Richards, B.K., 2005. Synthetic fibers as an indicator of land application of
sludge. Environmental Pollution 138 (2), 201–211.