pubs.acs.

org/journal/estlcu Letter

Per- and Polyfluoroalkyl Substances in Toilet Paper and the Impact

on Wastewater Systems
Jake T. Thompson, Boting Chen, John A. Bowden, and Timothy G. Townsend*
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ABSTRACT: Here, we evaluate a perhaps unexpected contributor

of per- and polyfluoroalkyl substances (PFAS) to our wastewater,
an input anticipated at every wastewater treatment facility�toilet
paper. In this study, both toilet paper and wastewater sludge were
Downloaded via UNIV OF FLORIDA on March 2, 2023 at 16:47:55 (UTC).

characterized to explore the magnitude of the potential PFAS

loading into wastewater systems from toilet paper. In both toilet
paper and wastewater sludge, 6:2 fluorotelomer phosphate diester
(6:2 diPAP) was the most prevalent PFAS detected, and toilet
paper usage was estimated to contribute from 6.4 to 80 μg/person-
year of 6:2 diPAP to wastewater−water systems. Our results
suggest that toilet paper should be considered as a potentially
major source of PFAS entering wastewater treatment systems.
KEYWORDS: 6:2 diPAP, biosolids, wastewater sludge, PFCA precursor, PFAS, paper

■ INTRODUCTION
With increasing frequency, scientists and policy makers rely on
toilet paper is often manufactured from recycled paper fibers
(which often can contain PFAS).21,22 We obtained packaged
toilet paper from four world regions (Africa, North America,
measurements from society’s wastewater as sentinels of disease
South and Central America, and Western Europe) and assayed
transmission,1 drug use,2 and chemical exposure and environ-
the concentration of 34 PFAS. Additionally, we analyzed the
mental release.3 Per- and polyfluoroalkyl substances (PFAS),
same suite of PFAS in sewage sludge from eight wastewater
ubiquitous chemicals now documented throughout the natural
treatment plants in Florida, United States (US), to comple-
environment,4 wildlife,5 human blood,6 and breastmilk,7 have ment the existing body of knowledge on PFAS in sewage
been similarly documented in wastewater,8 particularly the sludge, and these data, along with estimates of average toilet
residual sewage sludge from biological wastewater treat- paper usage and sludge generation, allowed us to assess the
ment.9,10 relative contribution of toilet paper to the PFAS loading, at
As PFAS occur in numerous household and personal care least for the dominant detectable species of PFAS, in
products,11,12 the measurement of PFAS in wastewater wastewater sewage sludge.
residuals comes as no surprise, but as much of the world’s
sewage sludge is land applied,13,14 the regulatory community
now wrestles with how best to address the human health and
environmental ramifications of PFAS discharges. Understand-
■ MATERIALS AND METHODS
Sample Collection and Extraction of Toilet paper.
ing the sources of PFAS and their respective contribution to Samples of toilet paper were collected from November 2021 to
wastewater will be key to prioritizing action�possible sources August 2022 by a volunteer network of students and
include the soaps, shampoo, and makeup that we wash down professors; individuals were given instructions to document
the location of toilet paper collection, the toilet paper recycled
the drain,11 clothes and related cleaning products in our
content, brand, and a picture or physical label of the toilet
laundry,15 and discharges from commercial and industrial
paper if possible (see Figure S1.). Samples were either
operations.16,17
collected as entire rolls or 10 g subsamples. Samples were
Here, we evaluate a perhaps unexpected contributor of
PFAS to our wastewater, an input anticipated at every
wastewater treatment facility�toilet paper. PFAS have been Received: February 8, 2023
documented in papers products18 and are a known wetting Revised: February 14, 2023
agent additive used to increase the efficiency of the pulping Accepted: February 15, 2023
process.19 The use of PFAS by paper mills has already been
identified as a source of environmental contamination20 and
could result in residual PFAS in toilet paper. Furthermore,

© XXXX American Chemical Society https://doi.org/10.1021/acs.estlett.3c00094

A Environ. Sci. Technol. Lett. XXXX, XXX, XXX−XXX
Environmental Science & Technology Letters pubs.acs.org/journal/estlcu Letter

Figure 1. Concentrations of 6:2 diPAP and relative distribution of detected PFAS in toilet paper. (A) Concentration of 6:2 diPAP in toilet papers
sampled from different world regions. (B) Concentration of 6:2 diPAP in toilet papers labeled as containing recycled paper compared to
nonrecycled toilet papers. (C) Proportion of each PFAS detected across all toilet paper samples relative to the total mass of ∑6PFAS. The
proportion of each PFAS is as follows: 91% (6:2 diPAP), 3.7% (8:2/6:2 diPAP), 1.3% (8:2 diPAP), 3.3% (PFHxA), 0.6% (PFOA), and 0.4%
(PFDA).

stored in sealed high-density polyethylene (HDPE) containers steel shovels washed with methanol prior to and between
for no more than 180 days at 25 °C. collections. Samples were transported to the laboratory,
Samples were extracted following a modified method aliquoted into 2 L HDPE bottles, and stored at −20 °C
published by Timshina et al.23 In short, triplicate 1 g until analysis. The sample extraction method is a modified
subsamples of toilet paper were transferred to 50 mL falcon version of a previously reported solid matrix extraction
tubes and spiked with 40 μL of an isotopically labeled internal method.24
standard mixture (see Tables S1 and S2). Extraction was Prior to extraction, wastewater sludge samples were
repeated three times in the following manner: after adding 10 homogenized by rotating for 20 min at 70 rpm and air-dried
mL of 0.3% methanolic ammonium hydroxide, samples were in a fume hood for a period of 3 days. A subsample of air-dried
agitated on a rotisserie rotator for 45 min, centrifuged for 10 wastewater sludge was taken to complete dryness at 110 ± 5
min at 4000 rpm, and supernatants transferred to 50 mL °C according to ASTM D221625 to determine the moisture
centrifuge tubes. Extracts were evaporated to approximately 1 content for calculating PFAS concentrations on a per dry mass
mL in a Biotage TurboVap before supernatants from the 3-fold basis. Air-dried wastewater sludge samples were extracted in
extraction were combined. For purification, combined extracts triplicate and divided into 10.0 ± 0.1 g subsamples, added to a
were vortexed for 30 s with 50 mg ±10 mg of ENVI-Carb 50 mL polypropylene tube, and spiked with 50 μL of an
graphitized activated carbon, centrifuged for 10 min, and isotopically labeled PFAS IS mixture. Then, 8.5 mL of 0.3%
transferred to new 15 mL tubes. The final extracts were ammonium hydroxide in methanol was added to the sample,
evaporated to 1 mL and aliquoted for analysis. Extracts were and the mixture was vortexed for 1 min, sonicated for 30 min,
stored no more than 30 days at −20 °C until analysis (see and rotated in an end-over-end fashion for 30 min. The sample
Table S4 for PFAS concentrations in toilet paper). Extraction was centrifuged at 4000 rpm for 10 min, and the supernatant
efficiencies were measured using pooled samples spiked with was removed using a pipet. An additional 8.5 mL of 0.3%
mass-labeled PFAS (additional information about the QC ammonium hydroxide in methanol was added to the remaining
process and reported extraction efficiencies can be found on sample, and the vortex, sonication, rotation, centrifugation, and
page S4 and Table S10 of the SI). supernatant removal steps were repeated. The combined
Sample Collection and Extraction of Wastewater extracts (17 mL) were concentrated to 10 mL under a gentle
Sludge. Wastewater sludge samples were collected from eight stream of high purity nitrogen gas (Biotage TurboVap II) at 35
facilities in Florida, US, between June 2021 and August 2021. °C. Extracts were then stored for less than 30 days at −20 °C
Samples were collected in 23 L HDPE buckets using stainless until analysis. Evaporated extracts were aliquoted into 200 μL
B https://doi.org/10.1021/acs.estlett.3c00094
Environ. Sci. Technol. Lett. XXXX, XXX, XXX−XXX
Environmental Science & Technology Letters pubs.acs.org/journal/estlcu Letter

Table 1. Fraction of 6:2 diPAP in Wastewater Sludge Expected to Come from Toilet Papera
Median 6:2 diPAP
concentrations in wastewater Average toilet paper usage Wastewater sludge generated Fraction of 6:2 diPAP in wastewater
Country sludge (ng/g) per capita (kg-person/year) per capita (kg-person/year) sludge attributed to toilet paper usage (%)
United 82 26 27 3.7
States27,31,32
Canada26,31,32 85 26 28 3.5
China30,31,33 12 7 28 6.9
Australia32,34,35 32 13 18 7.2
France29,31,36 1.7 10 21 89
Sweden28,31,36,37 5.6 15 25 35
a
The methodology for determining the fraction of 6:2 diPAP in wastewater sludge attributed to toilet paper usage is presented in the SI.

polypropylene autosampler vials for analysis by liquid that concentrations of 6:2 diPAP did not differ among regions
chromatography and tandem mass spectrometry (see Tables nor those identified with recycled content (p >. 05). These
S5−S8 for concentrations of detected PFAS in wastewater comparisons, along with the distribution of PFAS detected
sludge samples). among toilet paper samples, are shown in Figure 1 (A) - (C).
LC-MS/MS Analysis. Toilet paper and wastewater sludge Samples of wastewater sludge were collected and charac-
extracts were analyzed via targeted ultrahigh pressure liquid terized for PFAS to explore the magnitude of the potential
chromatography-tandem mass spectrometry (UHPLC-MS/ PFAS loading into wastewater systems from toilet paper.
MS) using a Thermo Vanquish UHPLC system (Waltham, DiPAPs were similarly among the most abundant PFAS
MA, US) coupled to a Thermo Quantis triple quadrupole mass measured in wastewater treatment sludges, though they
spectrometer (operated with electrospray in negative ioniza- represented a smaller contribution than in the toilet paper
tion mode and scanning via scheduled selected reaction itself. The sum of three diPAPs (6:2, 62:8:2, and 8:2 diPAP) in
monitoring (SRM)). the eight sludge sources tested contributed 54% ± 15% of the
A Gemini C18 (100 mm × 2 mm; 3 μm) column from ∑34PFAS, on average. Concentrations of 6:2 diPAP ranged
Phenomenex was used for chromatographic separation. Water from 27.2 to 750 ng/g with an average concentration of 141
[A] and methanol [B] both containing 5 mM of ammonium ng/g (PFAS concentrations can be found in Tables S5−S8 of
acetate were used as the mobile phases. The gradient elution the SI). Two studies on North American sewage sludge
was set as 0−3 min 10% B, 3−4.5 min 10%−35% B, 4.5−12.5 similarly identified diPAPs as the most abundant PFAS
min 35%−95% B, 12.5−12.51 min 95%−99% B, 12.51−19 measured with mean concentrations of 6:2 diPAP at 164 and
min 99%, and then equilibrated to initial conditions in 30 min. 121 ng/g.26,27 Interestingly, reported results from Europe and
The temperature of the autosampler was 4 °C. The flow rate China found diPAPs to contribute less to the total mass of
was 0.5 mL min−1, and injection volume was 10 μL. Water, PFAS present in wastewater sludge, with studies from Sweden,
methanol, and ammonium acetate used in the study were all France, and China reporting mean concentrations of 2.0, 2.9,
Optimal grade purchased from Fisher Scientific. Scheduled and 11.6 ng/g for 6:2 diPAP, respectively, which represented
SRM mode (monitoring two transitions, if possible) was used 3%, 2%, and 8% of the total mass PFAS measured.28−30
to detect and quantify each PFAS (native and mass-labeled The use of toilet paper, and whether it is flushed down the
species are shown in Tables S1 and S2 of the SI). The most toilet or disposed of with trash, varies regionally as a function
intense transition was used for quantification, while the second of cultural norms, economic means, and sanitation infra-
transition was used to confirm identification (if applicable). structure. Reported per capita toilet paper use in regions such
For PFAS where no isotopically labeled standard was available, as Japan, the US, Canada, and Western Europe ranges from 15
an alternative labeled standard, either similar by structure or to 25 kg/person-year, while estimated use in Latin America,
retention time, was selected for quantitation. Table S2 displays China, and Africa ranges from 2 to 10 kg/person-year.31 To
the mass spectrometric scan parameters (and transitions) for estimate per capita mass of 6:2 diPAP entering wastewater
all targeted PFAS, while Table S3 displays additional LC-MS/ systems from toilet paper, a mean 6:2 diPAP concentration of
MS parameters. Additional information regarding the QA/QC 3.2 ng/g from the 21 samples analyzed was applied to the
protocol, extraction efficiencies for each matrix, and data range of reported toilet paper usage rates. This results in an
analysis are available on pages S4−S5 of the SI. annual 6:2 diPAP contribution from toilet paper of 6.4 to 80

■ RESULTS AND DISCUSSION

Of the 34 PFAS analyzed, six were detected in the toilet paper
μg/person-year.
We next examined the contribution of toilet paper to the
total 6:2 diPAP found in wastewater sludge using region-
samples: perfluorohexanoic acid (PFHxA), perfluorooctanoic specific assumptions (concentration in sludge, per capita toilet
acid (PFOA), perfluorodecanoic acid (PFDA), 6:2 fluoro- paper use, and per capita sludge generation; Table 1). The
telomer phosphate diester (6:2 diPAP), 6:2/8:2 fluorotelomer estimated contribution from toilet paper was lowest in the US
phosphate diester (6:2/8:2 diPAP), and 8:2 fluorotelomer (3.7%) and Canada (3.5%) and highest in Sweden (35%) and
phosphate diester (8:2 diPAP). By far the largest single PFAS France (89%). The larger contribution observed in the two
among those detected was 6:2 diPAP, representing 91% ± 8% European data sets stems from the lower reported 6:2 diPAP
of the total mass of PFAS on average. A previous study sludge concentrations compared to those measured in North
identified elevated diPAP concentrations in paper mill wastes America (by an order of magnitude). Comprehensive PFAS
streams, suggesting that diPAPs could be introduced during data on regional wastewater slude sources are remarkedly
production.26 Analysis of variance (ANOVA) between regions, limited, but in those nations where multiple data sets were
and a t test between recycled and nonrecycled samples, found available (US and Sweden), each nation’s data were of the
C https://doi.org/10.1021/acs.estlett.3c00094
Environ. Sci. Technol. Lett. XXXX, XXX, XXX−XXX
Environmental Science & Technology Letters pubs.acs.org/journal/estlcu Letter

same magnitude (median of 7.0 and 1.8 ng/g in the Swedish Boting Chen − Department of Department of Environmental
studies and 53 and 100 ng/g in the US studies). Engineering Sciences, University of Florida, Gainesville,
Wastewater Implications. Our results suggest that toilet Florida 32611-6450, United States
paper should be considered as a potential significant source of John A. Bowden − Department of Department of Physiological
PFAS entering wastewater treatment systems. The lower Sciences, University of Florida, Gainesville, Florida 32610,
estimated 6:2 diPAP contribution derived from the North United States; orcid.org/0000-0003-3135-4015
American data sets, despite the greater reported use of toilet Complete contact information is available at:
paper in this region, suggests that other 6:2 diPAP sources, https://pubs.acs.org/10.1021/acs.estlett.3c00094
such as cosmetics,11 textiles, and food packaging,38 deserve
attention, especially given the apparently smaller contribution Funding
of these sources based on data sets from France and Sweden. This work was funded by the Hinkley Center for Solid and
Previous work from D’eon et al.26 suggests that diPAPs are a Hazardous Waste Management.
prevalent North American chemical contaminant. Perhaps
Notes
regional consumer product choices and discard practices can
The authors declare no competing financial interest.
inform strategies to reduce wastewater borne PFAS loads. This
reduction in PFAS is critical, since wastewater effluent and
sludge are commonly reused for irrigation and/or land
application;32 research has already shown that these two
■ ACKNOWLEDGMENTS
The authors would like to thank the Hinkley Center for Solid
pathways pose a risk for human and environmental exposure to and Hazardous Waste Management for providing the funding
PFAS.39 for this research. Additionally, the authors would like to thank
Complicating this discussion, however, is the fact that the all the students and researchers who contributed to the
dominant PFAS family observed in toilet paper and wastewater collection of toilet paper samples and give a special thanks to
treatment sludge, the diPAPs, are precursor species and have Francisca Hinz, Nicole Robey, and Dreyton Lott for their
the capacity to be transformed into terminal PFAS. Terminal contributions to the sample collection process.
species�such as PFHxA, PFOA, or PFDA�are formed from
the biologically mediated transformation of 6:2 diPAP and
other diPAP homologues,4,40 and these chemicals are the
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