Microplastics WHO Report (1)

Dietary and inhalation
exposure to
nano- and
microplastic
particles
and potential
implications for
human health
Dietary and inhalation
exposure to
nano- and
microplastic
particles
and potential
implications for
human health
Dietary and inhalation exposure to nano- and microplastic particles and potential implications for
human health
ISBN 978-92-4-005460-8 (electronic version)

ISBN 978-92-4-005461-5 (print version)
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CONTENTS
Acknowledgements v
Abbreviations and acronyms vii
Executive summary viii
1. Introduction 1
1.1 Background and scope 1
1.2 Definitions 3
1.3 Composition and properties of particles 5
2. Human exposure 13
2.1 Occurrence in drinking-water 13
2.2 Occurrence in air 20
2.3 Dermal exposure 26
2.4 Occurrence in food 28
2.5 Summary and recommendations 43
3. Observations from epidemiology 45
4. Dosimetry and biokinetics 49
4.1 Dosimetry: extrapolation from external to internal exposure 49
4.2 Biokinetics 57
4.3 Biokinetics: summary and recommendations 64
5. Toxicological effects 67
5.1 Literature review and experimental study evaluation 68
5.2 Nano- and microplastics as vectors of chemical exposure 84
6. Nano- and microplastics as vectors of pathogens 91
6.1 Microplastic-associated biofilms in water 91
6.2 Microplastic-associated biofilms in food 93
7. Summary and research topics 95
7.1 Summary 95
7.2 Options for curbing exposure 98
7.3 Key messages and research topics 98
References 101
Annex Quality assurance and quality control scoring for studies reporting
microplastic particles in air samples 137
FIGURES
Fig. 1. Attributes of nano- and microplastics to be considered in
assessing both exposure and hazard 6
Fig. 2. Classifications of manufactured and commonly encountered
plastic materials 7
Fig. 3. Concentrations of microplastic particles in drinking-water
according to particle size in studies with a total assessment
score ≥ 11 and in which particles were verified as plastic 19
Fig.4. Dietary consumption from 16 food categories derived from
all 17 GEMS/Food clusters 40
Fig. 5. Extrapolation of concentrations and doses of inhaled
particles in the respiratory tract of rats to humans 50
Fig. 6. Main regions of particle deposition in the human respiratory
system and modelled deposition of a 1-g/cm3 spherical particle in
relation to the diameter of the particle 52
Fig. 7. Approach used to evaluate studies of effects in vivo and in
vitro for use in assessing human health risks due to exposure to
nano- and microplastic particles 68
Fig. 8. QA/QC evaluation scores for 76 studies in mammals in vivo 70
Fig. 9. QA/QC evaluation scores for 76 studies in mammals in vivo
exposed by ingestion or inhalation 70
Fig. 10. QA/QC evaluation scores for 37 studies of effects in vitro 73
Fig. 11. QA/QC evaluation scores for 37 studies of effects in vitro
designed to reflect exposure by ingestion and by inhalation 81
Fig. 12. Uptake and biokinetics that influence the effects on human
health of exposure to nano- and microplastic particles 97
TABLES
Table 1. Average densities of commonly used polymers, the
applications of representative additives and the estimated typical
percentages added (weight/weight) of commonly used polymers 9
Table 2. Recent studies on the numbers and characteristics of
microplastic or microplastic-like particles in drinking-water 16
Table 3. Studies with a total assessment score > 10 of microplastic
particles in indoor and outdoor air at urban and rural sites 25
Table 4. Reported numbers of microplastic or microplastic-like
particles and particle characteristics in studies of their presence
in food and beverages for human consumption 31
Table 5. Estimated daily and annual per capita ingestion of
microplastic particles 38
Table 6. Criteria for evaluating the reliability and quality of in-vivo
and in-vitro studies on the effects and biokinetics of nano- and
microplastic particles 57
Table 7. Experimental study design criteria related specifically to
animal husbandry 76
BOXES
Box 1. Definitions of nano-, micro-, meso- and macro-plastic
particles and related terms 4
Box 2. Recommendations for improving sampling and analytical
methods 14
ACKNOWLEDGEMENTS
The World Health Organization expresses its appreciation to
all those who contributed to the preparation and development
of this report, including the colleagues named below.
This report is the product of several expert meetings held
between 2019 and 2022, and represents a follow-up to the
WHO report on Microplastics in Drinking Water, published in
2019.
Lead authors
• Alan Boobis, Imperial College London, United Kingdom
• Flemming Cassee, National Institute for Public Health
and the Environment, Netherlands
• Todd Gouin, Independent Consultant, United Kingdom
• Bart Koelmans, Wageningen University, Netherlands
• Shirley Price, University of Surrey, United Kingdom
• Sandra Wagener, German Federal Institute for Risk
Assessment, Berlin, Germany
• Stephanie Wright, Imperial College London, United
Kingdom
Experts who provided insights, wrote text, offered peer review,

and/or participated in meetings:
• Guillaume Duflos, Agence Nationale de Sécurité
Sanitaire de l’Alimentation, de l’Environnement et du
Travail, France
• John Fawell, Cranfield University, United Kingdom
• Jennifer De France, WHO, Switzerland
• Bruce Gordon, WHO, Switzerland
• Matthias Labrenz, Leibniz-Institute for Baltic Sea
Research, Germany
• Christine Lemieux, Health Canada, Canada
• Peter Marsden, Drinking Water Inspectorate, United
Kingdom
• Lidia Morawska, Australia-China Centre for Air Quality
Science and Management, Australia
• Maria Neria, WHO, Switzerland
• Sabine Pahl, University of Plymouth, United Kingdom
• Kim Petersen, WHO Switzerland
• Lisa Scheuermann, WHO, Switzerland
• Emanuela Testai, Istituto Superiore di Sanità, Italy
• Dick Vethaak, Deltares, The Netherlands
• Martin Wagner, Norwegian University of Science and
Technology, Norway
v
Dietary and inhalation exposure to nano- and microplastic particles and potential implications for human health
• Annemarie van Wezel, University of Amsterdam, The

Netherlands
The development and production of this document was
coordinated and managed by Lisa Scheuermann and Kim
Petersen (both WHO). Elisabeth Heseltine, France, edited the
report.
WHO also gratefully acknowledges the financial support
provided by the Ministry of Foreign Affairs, Norway.
vi
ABBREVIATIONS AND
ACRONYMS
ABS acrylonitrile butadiene styrene
bw body weight
EFSA European Food Safety Authority
FAO Food and Agriculture Organization of the United Nations
GEMS/Food Global Environmental Monitoring System – Food Contamination
Monitoring and Assessment Programme
LOD limit of detection
LOQ limit of quantification
MMAD mass median aerodynamic diameter
MOE margin of exposure
MP microplastic particles
NMP nano- and microplastic particles
NP nanoplastic particles
OECD Organisation for Economic Co-operation and Development
PBPK physiologically based pharmacokinetics
PCB polychlorinated biphenyl
PET poly(ethylene terephthalate)
PM particulate matter
PM10 particulate matter ≤ 10 µm
PM2.5 particulate matter ≤ 2.5 µm
PVC polyvinyl chloride
QA/QC quality assurance and quality control
QIVIVE quantitative in vitro to in vivo extrapolation
TAS total assessment score
WHO World Health Organization
wt weight
ww wet body weight
vii
EXECUTIVE SUMMARY
The number of reports of the presence of microplastic
particles (MP) in the environment has increased significantly
during the past few years. MP have been detected in air,
water, soil, food and beverages, indicating that exposure
of humans to these particles is ubiquitous. In 2019, the
World Health Organization (WHO) commissioned a report to
evaluate the evidence for risks to human health associated
with exposure to nano- and microplastic particles (NMP) in
drinking-water. The report was based on literature reviews
of studies published up to December 2021 in which original
data on the occurrence of NMP in air, water, food and
beverages were reported and also experimental studies
on their toxicity. WHO experts evaluated the quality of
the studies of environmental monitoring and of toxicity,
particularly with regard to the reliability and relevance of the
data for characterizing risk. The possible role of NMP as
vectors of chemicals and pathogens was also assessed, and
clinical observations from occupational epidemiology are
summarized. A key observation is that MP are ubiquitous in
the environment and have been detected in environmental
media with direct relevance for human exposure, including air,
dust, water, food and beverages.
There is increasing awareness of the occurrence of NMP
in air and their implications for human health. Studies
of the inhalation of NMP should include consideration
of their biokinetics, as their intake depends on their size,
shape, density and surface chemistry, which influence their
deposition in the alveolar regions of the lungs. Observations
from occupational epidemiology suggest that acute and
chronic exposure to elevated concentrations of NMP, such
as polyvinyl chloride dust and nylon flock, can result in harm
to the respiratory tract. Better characterization is necessary
of the properties of NMP in air, such as the fractions that
contribute to (regulated levels of) particulate matter and their
absolute concentrations. The current lack of such data limits
characterization and quantification of the impact of human
inhalation of NMP.
Ingestion of MP has been reported in a variety of foods and
beverages, including fish and seafood products, salt, sugar,
honey, rice, milk and drinking-water. Limited characterization
of the hazard of NMP due to dietary exposure suggests the
possibility of adverse outcomes similar to those of other
well-studied insoluble particles, as they have similar modes of
viii
Executive summary
action, including generation of reactive oxidation species and

stimulation of an inflammatory response.
A number of difficulties obviated an assessment of overall
human exposure to NMP, including the limited availability
of data on the occurrence of NMP measuring < 10 μm in
water, food and beverages. Observations from particle and
fibre toxicology indicate that particles < 10 μm are probably
taken up biologically. Most of the available studies on the
occurrence of NMP in water, food and beverages reported
particles measuring > 10 μm, which are unlikely to be
absorbed or taken up. As most of the toxicity studies were
conducted with a monodisperse group of plastic particles,
typically measuring < 10 µm, studies of effects and of
exposure are mismatched, obviating extrapolation of data on
toxicity for use in a quantitative risk assessment.
For this report, the quality, reliability and relevance of data on
both exposure and effects were assessed for their possible
contribution to a risk assessment of NMP. The assessment
scores indicated that the available data are of only very
limited use for assessing the risk of NMP to human health.
Several shortcomings were identified, the most important
of which was the heterogeneity of the methods used,
including use of “bespoke” methods for analysing data
obtained by environmental monitoring and inconsistencies in
observations of adverse effects. Assessment of the quality
of the studies should promote best practices in experimental
design to be used in future studies. It is generally
recommended that standard methods be developed and
adopted to ensure that the research community can reduce
uncertainties, strengthen overall scientific understanding and
provide more robust data for assessing the risks of exposure
to NMP to humans.
Research to improve exposure assessment, for instance,
should be designed to complement research on the dosimetry
and biokinetics of environmentally relevant NMP. Quantitative
data on the rate of translocation of NMP and on their size
distribution, shape, polymer composition and surface
chemistry in air, food and beverages, including drinking-
water, are necessary to determine which properties are
most relevant for studies of biokinetics and adverse effects.
This will require methods for characterizing and quantifying
NMP < 10 μm, depending on the media in which they are
dispersed.
The selection of in-vitro and in-vivo test systems should
be guided by better understanding of exposure. Testing
ix
of the toxicity of environmentally relevant NMP should

represent a priority, and it is recommended that a series
of well-characterized reference NMP be generated and
made available to the research community. Furthermore,
the applicability of tools for extrapolating results obtained
in vitro to the situation in vivo and physiologically based
pharmacokinetics models should be assessed and new
models developed as necessary.
Little is known about the adverse effects of MP-associated
biofilms, although the available data provide no evidence
of a risk to human health. NMP constitute only a fraction
of the particles that occur in the environment on which
microorganisms can colonize and form biofilms.
Additionally, little is known about exposure to NMP in air,
food and beverages, particularly with respect to the polymer
composition of NMP < 10 μm, the specific mass and the
identity of associated plastic additives, including factors
that might influence their bioavailability. It is therefore not
currently possible to characterize or quantify the potential
role of NMP in the transport of chemicals. The possibility of
enrichment of antimicrobial-resistance genes in
MP-associated biofilms and the role of NMP as vectors for
pathogens and chemicals should be studied further. Such
research would benefit from better measurement of exposure
made possible by the development and application of
standard methods.
Recommendations
Although the limited data provide little evidence that NMP
have adverse effects in humans, there is increasing public
awareness and an overwhelming consensus among all
stakeholders that plastics do not belong in the environment,
and measures should be taken to mitigate exposure to
NMP. This should include better management of plastics
throughout their product life-cycle and reducing the use of
plastics, when possible, to move towards a more sustainable
plastics economy. In addition to measures to better manage
plastic, such as innovations in waste treatment and initiatives
to reduce the use of plastics, innovations in materials science
should be supported, particularly to ensure substantial
reductions in the release of NMP from plastic products used
in commerce.
The weight of the scientific evidence provided by current data
on adverse effects of NMP on human health is low, because
of substantial limitations of the available information.
x
Executive summary
Strengthening of the evidence necessary for reliable

characterization and quantification of the risks to human
health posed by NMP will require active participation by all
stakeholders in developing and making available standard
methods. Standard methods should be developed to improve
the quality and reliability of data from both environmental
monitoring and studies of effects. Researchers should
ensure that studies on the sources and occurrence of NMP
in air, water, food and beverages are based on appropriately
designed, quality-controlled protocols.
Of particular interest are NMP measuring < 10 μm. Better
understanding of environmentally relevant exposure to these
particles could be used to generate reference NMP for use
in testing biokinetics and toxicity. Better understanding of
the toxicological effects of environmentally relevant NMP at
environmentally relevant concentrations, combined with use
of standardized toxicity testing methods will provide more
accurate dose–response relationships, from which threshold
effect concentrations can be derived and assessed for risk.
xi
1. INTRODUCTION
Both the intentional use and unintentional generation of nano- to micro-sized
plastic particles and their release to the environment have implications for human
and ecosystem health and are emerging public concerns. Increasing numbers of
studies have demonstrated the presence of nano- and microplastic particles (NMP)
in drinking-water, air, food and beverages, indicating possible risks to human health
associated with exposure to the particles and to chemical toxicants and biological
agents vectored by NMP (1–9).
WHO previously reviewed scientific information on microplastic particles (MP) in
drinking-water, drinking-water sources and wastewater (1, 2) to evaluate the potential
risks for human health. This report extends the assessment by including evaluations of
exposure to NMP from other sources, including the air and diet. Recent reports on MP in
food and the environment, such as those of the European Food Safety Authority (EFSA)
(10), the Science Advice for Policy by European Authorities (11), the Norwegian Scientific
Committee for Food and Environment (12), the Government of Canada’s Science
Assessment of Plastic Pollution (13), the Committee on Toxicity of Chemicals in Food,
Consumer Products and the Environment in the United Kingdom of Great Britain and
Northern Ireland (14) and the State of California (United States of America) (15), show
that the available data on exposure to MP and its effects are insufficient to conduct a full
quantitative risk assessment. This report summarizes current scientific understanding
of exposure to and the effects and potential risks of NMP in relation to human health
and provides guidance and recommendations for future research. The report:
• summarizes data on human exposure to NMP in food, beverages, drinking-water
and air and on the pathways specific to human health;
• examines the implications for human health on the basis of data on occurrence,
toxicology and exposure;
• when possible, identifies opportunities for mitigating exposure to NMP; and
• identifies gaps in the data and proposes topics for research.
1.1 Background and scope

As discussed in the WHO report on MPs in drinking-water
(1), plastic is considered to provide various social benefits.
For instance, it is a relatively inexpensive, flexible, robust,
lightweight, waterproof material that is easy to maintain
and sterilize and has insulating properties. The possibility
for moulding plastic into a wide range of shapes and forms
has led to its use in many commercial products, such as
packaging and building and construction materials, and in
the automotive and aerospace industries. Direct benefits
of the use of plastic to human health and the environment
include their role in extending the shelf-life of perishable
1
food items, their use in sterile medical devices, such as

examination gloves, syringes and intravenous tubes, and as
an inert material for prosthetics. The lightweight plastic used
in packaging and the automotive and aerospace industries
helps to reduce fuel consumption during the transport and
shipping of commercial goods and products. Furthermore,
the insulating properties of plastic used in construction
and electrical applications improve the energy efficiency
of housing and appliances, and the combined lightweight
and insulating properties of plastics help to reduce the
emission of greenhouse gases. Nevertheless, there is
growing awareness that unintended release of plastic to the
environment due to mismanagement of solid waste or lack
of infrastructure for efficient reduction, reuse or recycling of
plastic material at the end of its commercial life or daily use
can harm organisms, the environment and, potentially, human
health. By drawing attention to the prevalence of macro- and
micro-litter in the marine environment, researchers have
raised awareness of the negative effects of the release of
mismanaged solid plastic waste on marine organisms and
ecosystems.
Human exposure to NMP is intuitively linked to the
widespread use of plastic; however, identification of
the sources of NMP is a non-trivial exercise. Current
understanding is that humans are exposed to a complex
mixture of NMP of various shapes, sizes and polymer
composition from both primary and secondary sources. Data
for quantifying and characterizing exposure are limited. For
this report, we reviewed the available information on the
occurrence of NMP in food, beverages, drinking-water and
air. In view of the advancements in scientific understanding
of the effects of exposure to particles on human health, the
literature on the biokinetics and toxicity of particles in general
is also summarized.
Given the speed at which research on NMP is emerging
and the difficulty of conducting quantitative analyses in the
absence of standardized methods for assessing exposure
and effects, this report indicates how the available evidence
might be used to assess the implications of exposure to
human health by assessing the quality of studies on exposure
to and the effects of NMP, while also summarizing the
occupational and epidemiological data on the biokinetics of
exposure to particles. These various lines of evidence are
used to evaluate the implications of NMP for human health,
to recommend future research and to provide guidance for
mitigating exposure.
2
1. Introduction
Several reports on the effects of NMP on human health

published recently (1, 10–14) were key sources of information
for this report, as were results from the USA of a Southern
California Coastal Water Research Project workshop
organized in coordination with the State of California Water
Resources Control Board and the California Ocean Protection
Council to evaluate the human and ecological effects of
MPs in water (15–18). Data from several reviews were
also used, including on exposure to MP in food, beverages,
drinking-water and air (3, 5, 7–9, 19–23), epidemiological data
from studies of occupational exposure and the results of
experimental studies conducted in vitro and in vivo (24–27).
Although every effort was made to ensure that the literature
reviewed and evaluated for this report was as comprehensive
as possible, it is not possible in this rapidly emerging field
to guarantee that every study was retrieved. Additional
information was obtained by literature searches with various
online tools, including PubMed, Scopus and Google Scholar,
for studies published up to December 2021, and from external
experts who constructively peer-reviewed the report.
Studies with implications for human health on experimental
effects and environmental monitoring that contained
original data were evaluated and scored with respect to the
information they provided on various fundamental criteria
for quality assurance and quality control (QA/QC) (2, 3, 16,
28). The scores summarized in the report were not used
for screening or for prioritization but to provide a relative
indication of the reliability and relevance of the data for
determining whether exposure to NMP affects human health
and the results obtained contributed to recommendations for
strengthening the quality, reliability and relevance of future
studies. Details of the scoring of any individual study are
available upon request.
1.2 Definitions
The definitions, composition and properties of NMP have been debated for several
years. Below, we briefly summarize the terms commonly used in research on the
implications of exposure to NMP for the environment and human health. A common
definition of “microplastics” is plastic particles that are < 5 mm in diameter (11, 29,
30). This definition is perceived as a pragmatic approach for differentiating crudely
between macro- and microparticles in the marine environment (30–33).
A definition that is appropriate for assessing the potential effects of exposure to NMP
on the environment and human health, for both scientific and regulatory purposes,
remains, however, an area of debate (11, 29, 34–39). Contentious components of
defining NMP include the polymer composition and dimensions for differentiating
3
nano-, micro-, meso- and macroplastic particles. Nevertheless, regulatory bodies have
recently provided or proposed a number of definitions for regulatory decision-making.
The list in Box 1 is not exhaustive and is presented to illustrate various perspectives
and challenges associated with defining NMP. For a more thorough discussion, see,
for instance, references 37 and 38.
Box 1 Definitions of nano-, micro-, meso- and macroplastic particles

and related terms
Agglomerate – a collection of weakly bound particles or aggregates of which
the resulting external surface area is similar to the sum of the surface areas of
the individual components (40)
Aggregate – a particle comprising strongly bound or fused particles (40)
Airborne fibre – object with a length (L) > 5 μm, a diameter (D) < 3 μm and an
aspect ratio L:D > 3:1 (41)
Elastomer – a macromolecular material that returns rapidly to its initial
dimensions and shape after substantial deformation by a weak stress and
release of the stress (42)
Gas – a substance that (i) at 50 °C has a vapour pressure > 300 kPa (absolute)
or (ii) is completely gaseous at 20 °C at a standard pressure of 101.3 kPa (43)
Liquid – a substance or mixture that (i) at 50 °C has a vapour pressure of ≤ 300
kPa (3 bar), (ii) is not completely gaseous at 20 °C and at a standard pressure
of 101.3 kPa and (iii) has a melting-point or initial melting-point of ≤ 20 °C at a
standard pressure of 101.3 kPa (43)
Macroplastic – plastic ≥ 1 cm (25); plastic ≥ 5 cm (44); plastic litter ≥ 5 mm (45)
Microbead – plastic bead ≤ 5 mm used in products such as bath and body
products, skin cleansers and toothpaste (46); a microplastic used in a mixture
as an abrasive, i.e., to exfoliate, polish or clean (35)
Microfibre – a fibre of linear density approximately ≤ 1 dtex and > 0.3 dtex
(Note: dtex is a direct measure of the linear density of a fibre, as the mass of
fibre (g) per length (10 km) with a cross-sectional width of 5–10 μm (47, 48))
Microplastic – a material consisting of solid polymer-containing particles, to
which additives or other substances may have been added, and in which
≥ 1% w/w of particles have (i) all dimensions 1 nm ≤ x ≤ 5 mm, or (ii), for fibres,
a length of 3 nm ≤ x ≤ 15 mm and a length to diameter ratio of > 3. Polymers
that occur naturally and have not been chemically modified (other than by
hydrolysis) are excluded, as are polymers that are (bio)degradable (35); plastic
particle < 5 mm in diameter; 1 – < 1000 μm (37)
Nanofibre – a fibre with a cross-sectional dimension within a range of nm,
i.e., < 0.1 dtex or < 1 μm (47)
Nanomaterial – a material with any external dimension in the nanoscale or
with an internal structure or surface structure in the nanoscale (49); a natural,
incidental or manufactured material containing particles, in an unbound state
4
1. Introduction
or as an aggregate or an agglomerate, in which ≥ 50% of the particles in the

number size distribution have one or more external dimensions in the size range
1–100 nm (40)
Nanoplastic – plastic particle measuring 1 to < 1000 nm (37, 50)
Particle – a minute piece of matter with defined physical boundaries and a
defined physical boundary as an interface (40)
Plastic – a material that contains as an essential ingredient a high relative
molecular mass polymer and which, at some stage in its processing into
finished products, can be shaped by flow (42)
Polymer – a substance within the meaning of Article 3(5) of Regulation (EC)
No. 1907/2006 (51); a molecule of high relative molecular mass, the structure
of which essentially comprises the multiple repetition of units derived, actually
or conceptually, from molecules of low relative molecular mass (52)
Primary microplastic – intentionally produced plastic particles in the size
range 1 to < 5000 μm (36)
Secondary microplastic – microplastic particle in the size range 1 – < 5000 μm
formed by fragmentation in the environment during use (36)
Solid – a substance or a mixture that does not meet the definition of a liquid or
gas (43)
Synthetic plastic microbead – any solid plastic particle ≤ 5 mm intended to be
used to exfoliate or cleanse the human body or any part thereof (53)
Thermoplastic – material that can be softened repeatedly by heating and
hardened by cooling through a temperature range characteristic of the plastic
and, in the softened state, of being shaped repeatedly by flow into articles by
moulding, extrusion or forming. Note: Many thermoplastic materials can be
thermoset by appropriate treatment to induce cross-linking, e.g., by the addition
of a suitable chemical cross-linking agent or by irradiation (42).
Thermoset – plastic that, when cured by heat or other means, changes into a
substantially infusible, insoluble product (42)
1.3 Composition and properties of particles

Assessment of human exposure to NMP requires
characterization of several parameters, summarized in Fig. 1.
In addition to particle size, shape and polymer composition,
information on chemical additives and the physicochemical
properties of the polymer, such as surface activity and
particle density, is important for understanding their effects
on human health.
5
Fig. 1 Attributes of NMP to be considered in assessing both exposure

and hazard
1.3.1 Properties
The heterogeneity of NMP, which have various polymer compositions, sizes and
shapes, significantly complicates assessment of human exposure. The difficulty is
increased by the inconsistency in reporting of the properties of NMP in the scientific
literature. Furthermore, reporting of accurate data on each of the properties of NMP is
limited by analytical capability, which may vary significantly among research groups
(2, 3, 28, 54–60). Depending on the sample matrix, the sampling method may be
limited by the pore sizes of filters, which determine the lower size limit of particles that
can be sampled. When extraction and isolation of particles for analytical verification
are required, digestion methods may physically alter the size of the particles,
influencing quantification of exposure.
The analytical methods for verifying polymer composition are evolving constantly,
with the introduction of methods such as Fourier transform infrared spectroscopy,
Raman spectroscopy and pyrolysis–gas chromatography–mass spectroscopy. The
sensitivity of analysis with respect to both size and shape has improved, although
characterization of the polymer composition of fibres is still difficult (37, 39).
Laboratory studies on the effects of exposure to NMP on human health are often
limited to a single type of polymer and shape in a relatively narrow size range and may
not represent human exposure.
1.3.2 Composition
All polymeric materials can be divided into subclasses
according to the method of synthesis or some characteristic
of the material. For instance, manufactured synthetic
polymers can be classified into four main groups according
to their structure, origin or source, molecular force and
mode of polymerization (Fig. 2). In polymer science, for
6
1. Introduction
Fig. 2 Classifications of manufactured and commonly encountered

plastic materials
Source: adapted from references 64 and 65
instance, classification is based on structure (61–63). Other

aspects, such as the relative strength of the molecular forces
characteristic of the polymer, can strongly influence its
stability and its potential degradation and fragmentation to
NMP (64, 65).
Natural polymers such as cellulose, starch, proteins, rubber,
silk and wool, derived from plants and animals, may have
complex sequences of repeat units and physical properties
equivalent to those of synthetic polymers (62). A potentially
important question is whether the properties of synthetic
polymers make them more hazardous than natural polymers
and, if so, what those properties are.
The types of manufactured synthetic polymers to be included
as NMP is perhaps one of the most contentious issues
in defining NMP. There is no consensus on the inclusion
7
of elastomers, such as rubbers, or modified natural or

semi-synthetic polymers, such as rayon and cellophane,
in the definition (37, 39). A pragmatic approach has been
adopted for this report, which accounts for the complex,
heterogeneous nature of exposure to NMP. Consequently,
data on materials classified as synthetic polymers, which
include thermoplastics, thermosets, elastomers and fibres,
and have been reported in the literature as representative of
NMP are included. Fig. 2 lists plastic materials within each
polymer category that are typically encountered in commerce
and reported in the literature as contributing to NMP (12, 39,
62, 63, 66).
Synthetic polymers are used in plastic products because
of their properties (62). For instance, polystyrene has a low
thermal coefficient as a foam and has been widely used in
the production of coffee cups; poly(methyl) methacrylate
is a tough, transparent polymer and is used as artificial
glass; polyethylene is waterproof and provides a thin,
often clear, material for use in plastic films and food or
beverage containers; polyamide 66 (often referred to as
nylon 66) resists abrasion and degradation and is a semi-
crystalline thermoplastic produced as a monofilament and
used in textiles; poly(ethylene terephthalate) (PET) has low
permeability to carbon dioxide and is often used in soft-drink
containers; thermoset epoxies provide good adhesion and
strength and are thus used as adhesives; polyolefins have low
dielectric constants and are used in wire and cable insulation;
polyimides are resistant to high temperatures and are used
in electronics; and polyisopropene self-reinforces upon
extension and is used for its elastomer properties.
The composition of a polymer also influences its particle
density, which in turn influences exposure dosimetry in
toxicity testing. As discussed in section 4, the density of a
particle, with its size and shape, determine its deposition in
the respiratory tract. The densities of common polymeric
materials range between 0.8 and 2.3 g/cm3 (Table 1).
1.3.3 Additives
Few synthetic polymers are used commercially in the “pure” state. Some polyethylenes
and polystyrenes are sold as homopolymers without additives; however, chemical
additives and other materials are added to most polymers to improve their properties,
which can result in a range of densities (see Table 1) (62, 67, 68). On a weight basis,
fillers represent > 50% of all additives used, followed by plasticizers, reinforcing
agents, flame retardants and colouring agents (67). For instance, fillers such as finely
ground rubber are added to brittle plastic to add strength; composites of glass, carbon
8
1. Introduction
Table 1. Average densities of commonly used polymers, the applications

of representative additives and the estimated typical percentage
added (weight/weight) of commonly used polymers
Application
Density Flame Ultraviolet
Polymer (g/cm3) Anti-oxidant retardant Plasticizer stabilizer
Typical amount (% wt/wt) 0.05–3 0.7–25 10–70 0.05–3
Thermoplastics
Acrylonitrile butadiene styrene 0.98  
Polyamide 66 (nylon 66) 1.24    
Polycarbonate  
Polyethylene (amorphous) 0.85
• Low-density polyethylene 0.89   
• High-density polyethylene 0.96   

Polyethylene (crystalline) 1.00
Polypropylene 0.99  
Polystyrene 1.04    
Poly(vinyl chloride) (PVC) 1.39    
Thermosetting
Epoxide resin 1.2    
Phenol–formaldehyde resin 1.36  
Unsaturated polyester resin 1.23–2.3    
Polyurethanes 1.2    
Sources: references 1, 62, 67 and 68
or boron fibres are made for high-modulus and high-strength applications; and carbon
black and silicas are added to synthetic rubber formulations to resist tearing and raise
the modulus. Various plasticizers are added to lower the glass transition or reduce
crystallinity to soften the final product, such as in PVC. Polymeric properties can be
improved by adding silanes and other agents to improve bonding between the polymer
and other solid phases, such as glass fibres; both glass and rubbery polymers can be
cross-linked to improve elastomer behaviour or to control swelling.
Chemical plasticizers, an important group of plastic additives, can be added externally
to a polymer or internally, whereby they are chemically bonded. It has been estimated
that 80–90% of all plasticizers are used in a single polymer – PVC (67, 68). In the
absence of any plasticizer, PVC is a rigid solid, with limited commercial application.
Addition of a plasticizer helps to soften the polymer, and levels > 50% (wt %) have
been reported in applications such as shower curtains and vinyl upholstery (67).
Table 1 summarizes the levels of additives used in common polymeric materials.
9
The risks associated with use of chemical additives in commercial products are
usually assessed by the regulatory authorities responsible for authorizing their use in
commerce, including possible migration of additives into food from packaging (51,
69). Studies of the leaching of chemical additives into food from plastic packaging
were reviewed by Hahladakis et al. (68). Inhalation or ingestion of NMP after
degradation and fragmentation of plastic packaging, however, may represent a new
exposure pathway for chemicals of potential concern (68, 70–76).
1.3.4 Surface chemistry

The wide variety of solid plastic polymers results in
differences in the surface chemistry of particles. Depending
on the nature of the polymer, NMP are either charged
(positively or negatively) or neutral. In some applications, the
plastic may have a hydrophobic or hydrophilic coating, which
will further influence the fate of fragmented particles that
enter the environment. As the size of a particle decreases,
its surface area increases, increasing the number of surface-
expressed molecules and thus increasing the surface activity
of the particle (77). Reactive surface sites on a particle can
interact with protons and other ions in the environment, which
may also influence the toxicity of the particle. The surface
charge of a given particle is strongly influenced by pH and
ionic strength (in aqueous systems), which may influence the
extent of aggregation with other particles in the atmosphere
or water (78) – a consideration for toxicity testing.
Fotopoulo and Karapanagioti (79) observed significant
alteration of functional groups on the surface of eroded
polyethylene pellets. The formation of aldehydes, esters,
carbonyls, ketones and ketocarbonyls caused the surface
of weathered MP of polyethylene to be largely negatively
charged, whereas particles of weathered polypropylene
were reported to remain neutral. Differences in surface
chemistry may therefore occur according to the nature of the
weathering process and the composition of the polymer (64).
1.3.5 Particle size

The degradation and fragmentation of manufactured plastic materials to form micro-
and potentially nano-sized plastic particles are key factors in their implications for
human health. There is, however, no consensus on the upper and lower size limits
for differentiation of particles as microplastics and nanoplastics (36, 37). In the
definitions summarized in Box 1, MP are commonly defined as particles measuring
< 5 mm on the basis of a suggestion by Arthur et al. (29), which has been adopted by
the European Chemicals Agency (35) and referred to in the reports on microplastics of
the Science Advice for Policy by European Authorities (11) and WHO (1). Nevertheless,
10
1. Introduction
several groups have recommended that additional detail be added to the definitions of
particle size ranges, such as a lower size limit for MP of 1–20 μm (45, 50, 80) and an
upper size limit ranging from 500 μm to either 1 mm or 5 mm (45).
The size ranges of nanoplastic particles (NP) have also been debated. Gigault et al.
(50), for instance, suggested that NP are produced unintentionally by degradation and
fragmentation of plastic objects and show colloidal behaviour within the size range
of 1–1000 nm. Another common definition of NP is that of the European Commission
for nanomaterials (40), with an upper size of 100 nm. Alternatively, Hartmann et al.
(37) suggested an approach whereby nano- (1 – < 1000 nm), micro- (1 – < 1000 μm),
meso- (1 – < 10 mm) and macroplastic particles (> 1 cm) are differentiated, with
“particles” defined according to the definition of the European Commission in Box 1.
In view of the inconsistent terminology for specific particle size ranges, the term
“NMP” in this report refers to plastic particles measuring 1 nm to 5000 μm. Thus, MP
are particles measuring ≤ 5 mm, whereas NP measure ≤ 1 μm. In the environment,
the size of all plastic debris is distributed continuously, and characterization of the
distribution will simplify prioritization of the particle size categories that most strongly
influence human exposure (81). Particle size distribution and the limit of detection are
the basic parameters for interpreting information from environmental monitoring or
toxicity testing in assessing the implications of exposure to NMP on human health.
1.3.6 Particle shape

MP are typically reported to occur as spheres, irregular
particle fragments, fibres and films (1, 25). As the shape
of particles may be an important toxicological property
for human health effects, this information is reported
when available. In their assessment of particle shapes in
environmental samples, Burns and Boxall (82) reported
that the most abundant category in water and sediment is
fibres (48.5%), followed by fragments (31%), spherical beads
(6.5%), films (5.5%) and foams (3.5%). Similar distributions
of particle shapes were reported in fresh and drinking-water
(2). Kooi and Koelmans (81) estimated that, like particle
size distributions, particle shapes are also distributed
continuously, indicating the importance of characterizing the
shapes of NMP to ensure environmentally relevant testing.
• NMP are a heterogeneous mixture of particles and fibres of

various shapes, sizes, polymer composition, surface chemistry and
Key messages
associated chemicals.
• In this report, a pragmatic definition of microplastics is used, in
which synthetic polymeric particles are < 5 mm in diameter, while
NP are particles < 1 μm in diameter.
• The properties and composition of NMP change during their life-
cycle in the environment.
11
2. HUMAN EXPOSURE
Human exposure to NMP is widely recognized as occurring predominately through the
diet or by inhalation (1, 4, 10, 11, 19, 83, 84). The possibility of human exposure to MP
was raised by the observation of MP in seafood, such as mussels, intended for human
consumption (85, 86). Other studies have demonstrated the occurrence of MP in food
and drinking-water, food packaging and both indoor and outdoor air (1, 83, 87–94).
Thus, MP occur in drinking-water, a variety of foods and beverages and air, although
insufficient quantitative data are available for a full exposure assessment (95).
This section summarizes the data available for assessing human exposure to NMP.
A continuing challenge to characterizing and quantifying concentrations is, however,
the lack of standard analytical methods for identifying NMP of varying polymer
composition, size and shape in foods, beverages and air (2, 3, 11, 59, 60, 96–99).
Recommendations for improving sampling and analysis from the previous WHO
report (1) are shown in Box 2.
2.1 Occurrence in drinking-water

In the first WHO report (1), human exposure to MP in drinking-
water was summarized for both tap and bottled water. In a
critical review of nine studies of MP in fresh and drinking-
water, the concentrations ranged from 0 to 104 particles/L (1).
As mentioned above, characterization and quantification of
MP in drinking-water are limited by the lack of standardized
methods, which also limits comparison of studies, even by
the same group (2, 60, 100–103).
Koelmans et al. (2) proposed a scoring system for assessing
the quality of studies on concentrations of MP in drinking-
water. The system is based on eight criteria developed for
evaluating studies of MP in samples of biota (28) according
to how samples are collected, handled and analysed. A score
of 0, 1 or 2 is given to each criterion. The criteria are:
• description of the sampling method used,
• sample volume and number of samples,
• sample processing and storage,
• mitigation of contamination of samples during
preparation and handling,
• use of clean air conditions in the laboratory,
• use of negative controls to quantify and characterize
laboratory contamination,
• use of positive controls to quantify rates of recovery of
particles and
13
• analytical verification of polymers associated with

particles isolated from samples.
Box 2 Recommendations for improving sampling and analytical

methods
• Investigators should provide complete information about the method
used for sampling so that it can be reproduced.
• Sample volumes will depend on the nature of the matrix being sampled
and the size of the particles being analysed, which in turn are determined
by the filter or mesh size used. Sample volumes should be sufficiently
large to detect low MP concentrations reliably.
• Wherever possible, plastic material should not be used in sampling and
analysis. If plastic material must be used, it should be characterized and
reported.
• Materials should be rinsed with filtered water to avoid contamination.
• Sampling and sample processing should be done by trained
professionals, or the quality of samples collected or processed by
volunteers should be (quantitatively) validated against results obtained by
professionals.
• If preservatives are used, their effect on polymer mass or particle shape
should be evaluated, either in the study or from the literature.
• Laboratory surfaces should be thoroughly (wet) cleaned with filtered
water to avoid contamination.
• All samples should be handled in a laminar-flow hood or in a clean-air
laboratory.
• Blanks should be run, per day or per series, at least in triplicate, and
results should be corrected against blanks.
• Positive controls should be used to verify the recovery of particles during
digestion, density separation and filtration.
• Digestion should be used when necessary. Usually, digestion is not
necessary for drinking-water from a treated source. For more complex
matrices, such as foods and aerosol particles, however, in which high
concentrations of organic matter hamper the selection and (visual)
identification of particles, a digestion step is required.
• Polymers should be identified in a representative subsample of the entire
sample.
• Data should be reported as consistently as possible, such as number of
particles/L and mass/L for liquid matrices and particles/mass for solids,
with limits of detection for both number and mass concentrations and
minimum and maximum particle size. When possible, morphology should
be specified. Exposure concentrations must be reported consistently for
risk assessment.
14
2. Human exposure
• Standard methods should be developed for sampling and analysis, which

may depend on the media being sampled. For example, methods for
drinking-water will differ from those for foods and other beverages, as will
those for indoor and outdoor air. As far as possible, the same principles
should be followed.
The scoring system of Koelmans et al. (2) also includes an

assessment of sample treatment methods. As drinking-water
samples contain negligible amounts of organic matter, they
do not always require treatment, and a score of 2 is given
for sample treatment in such studies. A maximum total
assessment score (TAS) of 18 can therefore be attributed
to studies of drinking-water. The average TAS reported by
Koelmans et al. (2) are 13.7 (13–14) for studies of bottled
water, 12.5 (11–14) for those of drinking-water sampled at
treatment plants and 11.5 (8–15) for studies of treated tap
water. The study considered most relevant by Koelmans
et al. (2) for assessing human exposure is that of MP in
bottled water by Mason et al. (104), in which the average
concentration was 10.4 particles/L, consisting of larger
particles that could be confirmed as MP spectroscopically.
This study was chosen because it achieved a high
score on the criteria and reported the highest average
spectroscopically confirmed particle concentration; when
these elements are combined with other assumptions on
particle characteristics, the data represent a conservative
exposure scenario. Only four of the eight criteria, however,
were assigned a maximum score of 2, and Koelmans et al.
(2) recommended caution in using the data for a robust risk
assessment. Higher-quality data are necessary to improve
understanding of exposure to MP in drinking-water, a
recommendation echoed by Brachner et al. (96).
Since publication of the review by Koelmans et al. (2) and the
WHO report on MP in drinking-water (1), further studies have
reported on the presence of MP in drinking-water (Table 2).
These were obtained in a literature search in PubMed with
the keywords used by Koelmans et al. (2): microplastic AND
(bottled water OR tap water OR drinking-water).
15
Table 2. Recent studies on the numbers and characteristics of

microplastic or microplastic-like particles in drinking-water
Lower size Particles/L Particles/L Predominant Quality
Refer- bound in sample in blanks Particle size particle Predominant score
ence Water type (µm) (average) (average) (µm) shape polymer type (TAS)a
105 Bottled (mineral 1 384 Most No PET in plastic 13
water) particles < discussion bottles,
Glass 3074–6292 5 (> 75% in of shapes polyethylene
glass and > and styrene
Single-use PET 2649 95% in plastic butadiene
Reusable PET 4889 bottles) copolymer in
glass
106 Drinking-water 1 628 < 5% of ≤ 95% of Fragments, PET but also 11
treatment plant 338 counts in particles 1–10 closely polypropylene,
from surface 369 samples followed by polyethylene,
sources fibres polyacrylamide
(3 sites): raw,
treated
107 Bottled 5–20 14 ± 13 Typically, No PET but also 14
Single-use 14 40–50% in discussion polypropylene,
5–10 range; of shape; polyethylene
Returnable 118 > 80% < 20 described as
Glass 50 fragments
Beverage carton 11
104 Bottled 6.5–100 315 23.5 Not further No 14
lower specified characterization
bound
microscop-
ically and
in software
102 Tap from 10–100 0.2, 0.8 and Unknown Mainly Fragments PET, 14
groundwater 0.0 20–100 polypropylene,
sources (LOD, 0.3) ABS,
polyurethane
108 Tap from 20 0.0007 0.67 50–150 Fragments Polyester, PVC, 15
groundwater particles/L polyethylene,
sources 0.3 fibres/L polyamide,
epoxy resin
103 Tap from 24 60 Average not 0.5 Not specified Not specified No 9
sources reported, (LOQ = 4.1 characterization
as only one LOD = 0.9)
result
(5.5) > LOQ
104 Bottled > 100 10.4 4.15 Not further Fragments Polypropylene 14
specified (66%), (54%)
fibres (13%),
films (12%)
102 Tap from > 100 0.312 0.26 Not further Fibres (82%), PET, 14
groundwater (10-µm (LOD, 0.58) specified fragments polypropylene,
sources sieve size) (14%), polystyrene
films (4%)
91 Tap from 100 5.45 0.33 Fibre lengths, Mainly fibres No 8
unspecified (lowest particles/L 100–5000 (98.3%) characterization
sources reported) (5 particles
in 30 500-mL
blanks
109 Bottled 0.5 5.42 x 107 1 x 107 Range, Not reported Not reported 10
(estimated) 1.28–4.2
16
2. Human exposure
Lower size Particles/L Particles/L Predominant Quality

Refer- bound in sample in blanks Particle size particle Predominant score
ence Water type (µm) (average) (average) (µm) shape polymer type (TAS)a
110 Tap water 1 440 2.4 3–4453; > 50% Polyethylene 13
Mean, 66 fragments and
polypropylene
111 Drinking-water 1 6614 ± 1132 < 5% of 84.4–86.7% > 50% fibres PET, 12
treatment plant: (raw) samples 1–5 polyethylene,
raw and treated 930 ± 72 polypropylene
(treated)
112 Bottled 3 148 ± 253 0 ≥ 3 μm Not reported Not reported 15
113 Bottled 6.5 118 ± 88 ≥ 5 μm 62.8% fibres, Polyethylene, 14
14 ± 14 (returnable 37.2% polypropylene,
bottles) fragments PET
50 ± 52 ≥ 5 μm (single-
81.0 ± 3.0 15 use bottles)
≥ 5 μm (glass
26.0 ± 2.0 8 bottles)
12.0 ± 1.0 6 6.5–20 μm
20–50 μm
≥ 50 μm
100 Drinking-water 25 4.9 (raw) Depending > 25 Not ABS, 16
treatment 0.0011 on polymer reported polystyrene
plant: raw and (treated) composi-
treated tion. LOD,
1.1–65;
LOQ,
3.3–197
114 Tap water 500 18 ± 7 1 50% < 0.5 mm; Fibres Poly 14
(range, 25% 0.5–1 (trimethylene
5–91) mm terephthalate);
epoxy resin
115 Tap water from Not 2.8 Not Not reported Fibres Polyethylene 6
groundwater defined reported
wells
116 Tap water 10 0.7 ± 0.6 Not ≥ 500 μm Fibres PET and rayon 11
(range, reported (> 50%) (99.2%)
0.3–1.6)
117 Tap and bottled 25 2.1 ± 5.0 Not 25–500 Not Polyethylene, 10
water (range, reported reported polystyrene,
0.99–26) PET
118 Tap water 50 0.6 (range, Not ≥ 100 μm (> Fragments Polyethylene, 11
0.24–1.00) reported 80%) (77.5- PET,
93.6%) polypropylene
119 Tap water 1 343.5 1-7 1–10 (85%) Fragments Polyethylene, 11
polypropylene,
PET
120 Bottled 15 6–58 Not 40–723 Fragments Not reported 6
reported and Fibres
121 Tap water Not 39 ± 44 Not 19.2–4200 Fragments polystyrene, 11
defined (range, reported styrene-
1.9–225) ethylene-
butylene,
polypropylene,
polyester
122 Groundwater Not 38 ± 8 Not 18–491 Fragments Polyethylene, 15
defined (range, reported 94%) polypropylene,
16–97) PVC, nylon, PET
123 Tap water 13.23 < 5% 100–200 Fragments Nylon, 9
polyester
17
ABS, acrylonitrile butadiene styrene; LOD, level of detection; LOQ, limit of quantification
a
TAS, total accumulated score. The maximum score is 18. The score is calculated by adding the scores for
nine quality criteria; for each criterion, a score of 0, 1 or 2 is assigned. TAS values are shown in bold when all
the underlying scores are non-zero.
Ball et al. (100) reported use of a robust protocol for quality assurance and quality
control when measuring MP in drinking-water (i.e., tap water) at several sites in the
United Kingdom, and the data were evaluated as being of relatively high quality, with a
TAS of 16. Consistent with concerns raised by Koelmans et al. (2) regarding potential
laboratory contamination of samples during handling and preparation, Ball et al. (100)
reported relatively high, variable contamination of blanks, particularly for MP ≥ 25 μm.
In line with the recommendations of the Association of Official Agricultural
Chemists, an internationally recognized body that makes recommendations for
quality assurance and quality control, both a limit of detection (LOD) and a limit
of quantification (LOQ) are calculated for each polymer verified as representing
MP ≥ 25 μm. The LOD ≥ 25 μm is defined as the mean of blank samples plus 3.3 × the
standard deviation of the blank, whereas the LOQ ≥ 25 μm is calculated as the mean of
the blank samples plus 10 × the standard deviation of the blank. In estimating both the
LOD ≥ 25 μm and LOQ ≥ 25 μm, 10 blanks were analysed with drinking-water samples.
The concentrations were reported to be typically below the LOQ ≥ 25 μm, with only
three observations at 0.0008–0.002 particles/L; 20 samples were above the
LOD ≥ 25 μm, at 0.0004–0.0041 particles/L. The polymer types identified most
commonly as MP ≥ 25 μm in drinking-water were polystyrene and acrylonitrile
butadiene styrene (ABS). In the assessment of raw surface water, sampled before
treatment for use as drinking-water, the density of MP ≥ 25 μm was reported as
about 15 particles/L, the highest concentration being 113 particles/L (100). The
concentrations in surface waters are consistent with the range of values reported by
WHO (1) and are significantly higher than those in drinking-water, indicating efficient
removal of MP ≥ 25 μm during treatment of raw surface water (> 99.99%). The types
of polymers in raw surface water and drinking-water differ, MP ≥ 25 μm in raw surface
waters consisting mainly of polyethylene, PET and polypropylene and that in drinking-
water being mainly polystyrene and ABS. This observation suggests a source of MP
after treatment, i.e., from within the drinking-water treatment, storage and distribution
system (100, 108, 118, 119, 123).
Although most studies of MP in drinking-water continue to be conducted in Europe,
observations have also been reported from China (110, 111, 116, 119, 123), Mexico
(114), Thailand (113, 118), Saudi Arabia (117), Australia (122) and various other
locations, such as Japan and the USA (121). The concentrations in tap water were
generally consistent at all locations, ranging from below the LOD to 1247 particles/L
(Table 2). Exposure from bottled water may be more variable, concentrations as high
as 5.4 × 107 having been reported (109). In view of the variation in concentrations, the
data reported in Table 2 could be used in a probabilistic quantitative assessment of
human exposure in relation to drinking-water (124). When appropriate, exposure could
be reduced by use of technologies to treat both wastewater, to limit the release of MP
into the environment, and drinking-water, to reduce the concentration of particles (1,
100, 111, 119, 125).
18
2. Human exposure
As noted above, the previous WHO report (1) concluded that the data of Mason et al.
(104), with other assumptions on particle characteristics, represent a conservative
exposure scenario, and they were used to estimate exposure to chemicals that
might be associated with MP in drinking-water (1). Studies published more
recently provide additional information on the concentrations of MP in drinking-
water according to particle size distribution, allowing re-evaluation of the previous,
conservative exposure scenario. Kankanige and Babel (113), for instance, reported
concentrations of MP measuring 6.5–50 μm in 10 brands of single-use PET-bottled
water in Thailand. The particles were reported to consist mainly of fibres measuring
6.5–20 μm at concentrations of 29–127 MP/L. Their observation that the number of
particles increases with decreasing particle size is consistent with those of others,
such as Ossmann et al. (105), Schymanski et al. (107) and Winkler et al. (112).
Consequently, studies that include particle concentrations in relation to particle size
distribution, with verification of the polymer composition of MP, are perceived as
informative for exposure assessment (Box 2). Fig. 3 summarizes the concentrations
of MP in drinking-water found in the studies listed in Table 2, with the approximate
concentrations of the particle sizes reported.
Fig. 3 Concentrations of MP in drinking-water according to particle size

in studies with a total assessment score ≥ 11 and in which
particles were verified as plastic
The lower box indicates the 25th percentile, the black line indicates the mean, and the
upper box indicates the 75th percentile. The whiskers above and below the box indicate
the maximum and minimum values, respectively. The numbers of data points for each
particle size range are: 3 (1–5 μm), 5 (5–10 μm), 5 (10–20 μm), 6 (20–100 μm) and 5 (100–
500 μm).
19
Fig. 3 illustrates the general trend to increasing concentration with decreasing particle
size, with relatively high, variable concentrations of MP in drinking-water reported to
measure between 1 and 5 μm. The data do not include concentrations of particles
not confirmed as MP, including those < 10 μm reported by Mason et al. (104) and
Zuccarello et al. (109), as concern has been raised about whether all these particles
are MP (112, 126).
The analytical difficulty of verifying the polymer content of particles < 5 μm should
be emphasized, particularly in communicating results for use in assessing human
exposure (126). Assessment of human exposure to MP should therefore be based
only on studies that transparently and robustly adhere to the quality criteria defined
by Koelmans et al. (2). Research to ensure robust analysis of NMP < 5 μm in drinking-
water is thus critical, as these particles may be of greater concern for human health
than particles measuring > 5 μm.
2.2 Occurrence in air

2.2.1 Particulate matter in air
Particulate matter (PM) in air is a complex mixture of
particles from various sources, generated both naturally
and by human activity (127, 128). NMP in air are thus one
component of a heterogeneous mixture of particles. The
concentrations can be characterized in various ways,
most commonly as mass per volume but also as particle
number counts or total surface area per volume of air. “Total
suspended particulate concentration” was once a routine
metric for monitoring PM in air, covering a broad particle-
size range distribution of 0 to about 40 μm. For the purposes
of human health risk assessment, respirable PM are often
defined as particles with an aerodynamic diameter of
< 2.5 μm (fine particles) (127). The “inhalable fraction” refers
to coarse particles with an aerodynamic diameter > 2.5 μm,
which are usually defined as the fraction between 2.5 and
10 μm, although references to the “inhalable fraction” can
include sizes ≤ 100 μm (127–129). Aerodynamic diameter is
used as a surrogate for particle size. Ultrafine particles have
a mobility diameter < 0.1 μm and do not usually contribute
significantly to the total mass of particles; however, when
expressed as particle number counts, particles < 0.1 μm
dominate the entire respirable size range. The contributions
of different sizes of PM to airborne particle mass (or number)
vary substantially, as do the emission factors, including
gaseous precursors, physical characteristics and chemical
composition. Studies of the contribution of particles due
to tyre and road wear to PM ≤ 10 µm (PM10), for instance,
indicated an average contribution of about 1.9%, with a range
of 0.42–2.48%,
20
2. Human exposure
Coarse particles (i.e., 2.5–10 μm) are usually formed by

mechanical degradation of larger solid particles, the amount
of energy required to degrade the particles into smaller sizes
increasing with decreasing particle size (132, 133). Thus, in
urban environments, coarse particles are usually associated
with dust from roads and industrial activity but may also
include biological material such as pollen grains and bacterial
fragments. In coastal areas, evaporation of sea spray can
contribute to the composition of coarse particles. Fine and
ultrafine particles, i.e., < 2.5 μm aerodynamic diameter,
are formed mainly from gases and are generated during
combustion and degradation of material.
2.2.2 Nano- and microplastic particles in air

While concern about human exposure to MP in drinking-water, foods and beverages
has increased, awareness is also emerging of the presence of MP in air (20, 25, 134-
141). Like those in surface waters, the concentrations of MP in the atmosphere are
probably subject to spatial and temporal variation, as emissions to the atmosphere
are influenced by differences in the size of sources (such as between industrial or
urban and remote locations) and in processes that influence atmospheric transport
and mobility (i.e., wet and dry deposition, air speed and direction) (20, 142–147).
The main sources of MP in air have been considered to be a combination of
degradation and fragmentation of textiles in indoor air (87, 88). Tyre and road wear
particles are common in urban outdoor air and are generally characterized as
agglomerations of smaller particles from tyre wear and road dust (131, 143, 145, 146,
148). In the context of human health, however, atmospheric particles (section 2.2.1)
are complex, heterogeneous mixtures of solid particles and liquid droplets of varying
shapes and sizes that are present in the air we breathe, both indoors and outdoors.
In studies of human exposure by inhalation, particle size should be characterized
in order to differentiate between respirable and inhalable fractions. As discussed in
section 2.2.1, the fraction of tyre and road wear particles that contributes to PM10
can range from 0.42 to 2.48% on a particle mass basis and that of PM2.5 (respirable
fraction) may be < 1%. Particle size is currently used for regulatory purposes to control
PM mass concentrations on the basis of statistical relations between the size of
PM and implications for human health and is also the metric usually used in both
epidemiological and toxicological studies. Therefore, given the uncertainty about the
effects of exposure to NMP on human health, regulatory guidance on PM might be
considered to address this concern (9). Such guidelines could be used in a preliminary
default position, as regulatory values such as those set by WHO (133) are based on
the total mass of PM and do not differentiate the potential toxicity of the components
of PM. Characterization of the size and shape of NMP in both indoor and outdoor air
could then guide the design of mechanistic studies of toxicity, preferably supported by
appropriate data on biokinetics and biodynamics.
21
Characterization of NMP in air can thus contribute to overall understanding of the

heterogeneous mixture of PM. The chemical characteristics of the main constituents
of both PM10 and PM2.5 are known to be dominated by organic carbon and nitrate
(129, 132, 133, 149, 150). PM in air can be characterized and quantified by a variety
of methods in which flow rates can be actively pumped from low to high volume. As
reviewed and summarized by Enyoh et al. (20), the methods depend on the target
contaminant and research question. Importantly, for NMP, none of the methods
currently available allows online monitoring for discriminating and differentiating
NMP from other types of PM. Air samples must therefore be collected and analysed
retrospectively.
The available methods for sampling air for PM are based on gravimetric
measurements, which may be the most appropriate for assessing the concentrations
(on a mass or particle count basis) of MP in air. They include low- and high-volume
air samplers fitted with a size-selective inlet head (i.e., PM10), a filter substrate and a
regulated flow controller (e.g., Kleinfiltergerat and Partisol samplers). Another widely
used sampling instrument is the “tapered element oscillating microbalance”, in which
a sample is collected onto a filter and real-time short-resolution gravimetric data are
provided, although this feature is useful only if microplastics concentrations are to
be assessed for short-time resolution. Standard guidelines are available for mass-
based PM measurements, such as the European Committee for Standardization EN
12341, which could be used to contextualize the concentrations of microplastics in
air and determine their relative mass contribution to total PM or on a particle count
basis. Other instruments for characterizing and quantifying MP in air include cascade
impactors, which are used to collect and separate PM on impaction substrate filters
or discs by aerodynamic size distribution, and cyclone samplers, which give results
that can also be interpreted on a mass or particle count basis. The sampling duration
in the various methods depends on the volume of air sampled and the level of PM in
the air. When sensitivity is not a concern or longer sampling is required, low-volume air
samplers (~ 17 L air/h) are appropriate; however, when sensitivity is important, larger
volumes of air are to be sampled or shorter temporal resolution is required, high-
volume air samplers (~ 70 m3 air/h) are more appropriate (3).
Complementary use of outdoor air monitoring and modelling is effective for deriving
estimates of the exposure of the general population to PM. Exposure estimates can
be refined by use of portable personal samplers, which indicate individual exposure
from both outdoor and indoor air, or use of low- and high-volume air samplers with
conventional impactor or cyclone technology, which provide integrated daily, or longer,
measurements of size-separated PM fractions (3).
All sampling instruments risk being contaminated by their plastic components and
by external variables of background contamination and human error. Systematic
collection of blanks is therefore recommended, such as those collected with samples
in the field, travel blanks to assess contamination during transport and blanks to
evaluate background contamination during analysis and handling of samples (3,
139). Filter-based sampling requires downstream compositional analysis. For MP,
this is usually done with operator, semi-automated or automated micro-spectroscopy
(Raman or Fourier transform infrared) (3, 139). Whether samples are analysed directly
on the filter or extracted and processed first, the filter substrate must be carefully
22
2. Human exposure
selected to avoid analytical signal interference or disintegration during processing,

such as in the case of organic substrates. Liquid impingers eliminate the requirement
for a sample substrate, reducing the handling time of samples. A disadvantage,
however, is that evaporation may occur during sampling and should be assessed.
Viscous, non-evaporative liquids such as mineral oil are thus recommended for
collecting samples, although these types of liquids might interfere with downstream
analysis.
In view of recent interest in characterizing and quantifying MP in air, several
uncertainties in method development should be addressed, such as reporting on the
aerodynamic size distribution of MP in air and the most appropriate concentration
units for reporting data. The size distribution of MP must be evaluated, for instance, in
order to choose the appropriate tools, as some instruments are suitable for measuring
total suspended particulate and others for differentiating particle size fractions. If
the size fractions of the MP have implications for human health, larger monitoring
programmes may be warranted to evaluate their spatial and temporal variation or to
assess personal exposure. Differences in exposure to microplastics during different
activities and in different environments, such as indoors and outdoors, should be
considered. The concentration units used can significantly affect how samples are
collected, processed and analysed. If data are to be reported as particle counts, the
particles must be isolated from samples and examined visually without destroying
them. Mass concentrations from particle size impactor samplers might allow
quantification of the particle size distribution of NMP in air but would probably not
allow characterization of particle shapes.
Most counts of MP in air have been made by analysing bulk deposition (3, 139). These
results do not, however, necessarily correspond to the inhalable fraction relevant
for assessing human exposure or to estimated exposure, which is in a volume of
air. Exposure to MP by inhalation was studied by Liu et al. (151), who estimated that
inhabitants of Shanghai, China, were exposed to 21 particles/day by inhalation. The
size of the particles 1.7 m above the ground was reported to be 23.07–9955 μm, with
an average of 597.5 μm. While particles > 30 μm in aerodynamic diameter are less
likely to enter the nasal passages (152), these data provide an indication of human
exposure to MP in air. Ingestion may also be relevant. For instance, particles > 30 μm,
which are less likely to be inhaled, may be deposited on food or be ingested with dust.
Studies with better differentiation and quantification of exposure to MP < 30 μm, and
particularly PM2.5 and PM10, would refine assessments of exposure by inhalation.
Wright et al. (3) recently reviewed and evaluated 27 studies reporting MP in air and
atmospheric deposition for their reliability by the approach described by Koelmans et
al. (2) for drinking-water. Briefly, the studies were scored for 11 criteria. As for studies
of drinking-water, a value of 2 (reliable), 1 (limited reliability) or 0 (unreliable) was
assigned for each criterion. The final score is expressed as a TAS, calculated as the
sum of each score, to a maximum value of 22. According to Hermsen et al. (28) and
Koelmans et al. (2), a reliable study should have no 0 scores for any criterion. The only
study for which there were no 0 scores is that of Wright et al. (153), who reported MP
in atmospheric deposition collected in London, United Kingdom, during January and
February 2018. They reported a predominance of fibres with diameters of 5–75 μm,
most being 400–500 μm long. The most abundant non-fibrous particles measured
23
75–100 μm, the smallest particle being 25 μm in diameter, the smallest particle that
could be identified with the method used. The deposition rates were estimated to be
575–1008 MP/m2 per day. The particles reported by Wright et al. (153) were
> 25 μm, which are likely to be deposited in the upper airways and be swallowed,
implying ingestion.
Summaries of the TAS for all studies are reported by Wright et al. (3). In the interest
of brevity, the information presented in this report (Table 3) is thus limited to that
from studies of MP in air with a TAS > 10, which are relevant to human exposure via
inhalation and in which concentrations in air of MP < 20 μm are reported. It should
be noted that discrimination of particles < 50 μm was poor in all the studies reviewed
by Wright et al. (3), most studies reporting a predominance of particles > 50 μm. For
instance, the average size of non-fibrous particles was 164 ± 167 μm, and particles
measuring 75–100 μm were the most abundant. Gaston et al. (154) reported that 30%
of fibres measured 100–300 μm, with an average particle fragment size of
104 μm in outdoor air and 58.6 μm in indoor air; Wang et al. (155) reported an average
particle size of 851 μm; Liu et al. (151, 156) reported averages of 582 μm and
246 μm; and Dris et al. (135) reported that most of the fibres detected were
200–400 μm in length. Some authors, such as Allen et al. (142) and Bergmann et
al. (143), found that particles < 50 μm predominated; however, the distribution of
particles < 50 μm remains uncertain, and additional data are necessary to estimate
the abundance of particles most relevant for inhalation, i.e., < 20 μm (Table 3).
Generally, the distributions reported are limited by the analytical method used, and
the microscopic magnification used strongly influences the results. Use of higher
magnification and an appropriate analytical method, such as Raman microscopy,
would allow detection of smaller particles. The variations in particle size reported
therefore do not necessarily reflect the actual particle size distribution in air but may
be artefacts of the analytical method used.
In some studies, fibres with diameters of 5–75 μm predominated, while in others
mainly fragments measuring < 10 μm to > 2 mm were found, and mainly particles
< 50 μm were found in others (Table 3). The shapes and sizes are influenced by
factors including inconsistencies in sampling, sample preparation and analysis.
Standard methods should be developed for accurate characterization and
quantification of MP in air (3).
To characterize human indoor exposure by inhalation, Vianello et al. (159) collected
air samples from three apartments in Aarhus, Denmark, with a “breathing thermal
manikin”, which simulates the presence of a human occupant and has an inlet at the
mouth connected to a low-volume air pump. Samples were collected on filters that
were then analysed by micro-Fourier transform infrared spectroscopy, with a lower
instrument size LOD of 11 μm. The results suggested that MP were ubiquitous in the
air, at concentrations of 1.7–16.2 particles/m3, consistent with Gaston et al. (154)
but significantly lower than those reported by Liao et al. (157). The most abundant
polymers were polyester (59–92%), polyethylene (5–28%), nylon (0–13%) and
polypropylene (90.4–10%); the concentrations of non-synthetic particles were one or
two times higher (159). In contrast to studies that reported a predominance of fibres,
the studies summarized in Table 3 suggest the relative importance of fragments
in human inhalation exposure. As most people spend much of their time indoors,
24
2. Human exposure
Table 3. Studies with a total assessment score > 10 of microplastic

particles in indoor and outdoor air at urban and rural sites
Lower Number of
size Particle particles Predominant
Refer- bound concentration in blanks particle Predominant Quality
ence Sample type (µm) (average) (average) Particle size (µm)a shape polymer type scoreb
157 Air (indoor 10 1583 ± 1181 3.3 ± 1.8 > 90% < 100 μm, Fragments Indoor: 18
and outdoor) MP/m3 (indoor); MP/filter the 5–30 μm > 80% Polyester,
189 ± 85 MP/m3 fraction polyamide,
representing polypropylene
54.1–65.2% of total Outdoor:
Polyethylene,
polystyrene,
polyester
158 Air (outdoor) 10 282 ± 127 MP/ 3.9 ± 2.2 73.5–96.6% Fragments Polyethylene, 18
m3 (range, MP/filter represented by (88.2%) polyester,
104–650) 5–30-μm fraction polystyrene
159 Air (indoor) 11 9.3 ± 5.8 7.7 ± 3.8 36 and 21 for the 13% fibres, 81% polyester, 15
particles/m3 MP/blank, major and minor 87% 6% polyethylene,
data not dimensions fragments 5% polyamide,
corrected 2% polypro-
pylene and 6%
other polymers;
non-synthetic,
95% protein,
5% cellulose;
MP > 4% total
particles
154 Air (indoor 20 3.3 fibres and 2.4 fibres Fibre lengths: Fragments Polystyrene, 15
and outdoor) 12.6 fragments/ and 12.2 indoors 641 μm; PET,
m3 indoors; 0.6 fragments outdoor 616 μm; polyethylene
fibres and 5.6 per filter approx. 30% of
fragments/m3 indoors; fibres 100–300 μm
outdoors 0.4 fibres Fragments:
and 6.3 outdoors 104 μm;
fragments indoors 58.6 μm
per filter
outdoors
156 Air (outdoor) 12 0.41 MP/m3 No MP in 246.52 μm (12.35– Fibres (43%), PET (51%), 13
(0–2 MP/m3) one blank 2191.32 μm) fragments epoxy resin
collected (48%), beads (19%),
(9%) polyethylene
(12%), alkyd
resin (8%);
fibrous PET
(87%)
142 Deposition 10 365 ± 69 3±1 Most MP < 50 Fragments > Polystyrene 12
particles/m2 fibres, 1 µm, fibre lengths fibres > films (as fragments)
per day (range, ± 1 film predominantly followed by
29–462) and 8 ± 1 100–200 μm and polyethylene
fragments 200–300 μm (max,
per filter 3000 µm); films,
50–200 μm
Additional information presented by Wright et al. (3) and in Annex 1

a
Geometric mean
b
Maximum, 22
25
inhalation of indoor air is probably an important exposure pathway; however, in view

of the LOD of 10 μm, inhaled particles are unlikely to reach the alveolar regions of
the lungs and are more likely to be swallowed (section 4). Thus, methods should be
developed for monitoring MP < 10 μm.
The data summarized in Table 3 indicate the ubiquity of MP in both indoor and
outdoor air; however, differences in sampling methods and in the reporting of results
prevent comparison of the data with air quality guidelines for PM or with effect
thresholds reported for polymeric particles in occupational exposure. Inconsistent
units and lack of standardized sampling methods obviate comparisons of studies and
extrapolation. The data do imply, however, that MP in air are a potentially important
source. As none of the methods used allows quantification of concentrations
< 10 μm, limited information is available on exposure to the respirable fraction of NMP,
< 2.5 μm. The particle sizes in deposition studies reviewed by Wright et al. (3) indicate
whether exposure occurred via ingestion, directly, as dust or as dust settled on food
during its collection, manufacture, packaging, distribution and preparation. Limited
information is available on contamination of food with MP (160). Given the ubiquity
of MP in atmospheric deposition, research should be conducted to determine overall
exposure in air and the stages at which NMP are introduced into food.
Because of the lack of data on the contribution of MP with PM10 and PM2.5, human
exposure to the respirable fraction of MP cannot be estimated reliably. In view of the
quality score for characterizing deposition of MP in the study of Wright et al. (153), the
value of 771 MP/m2 per day could be used as a conservative estimate of deposition
of particles measuring 100–500 μm onto surfaces. The value of 9.3 ± 5.8 MP/m3
reported by Vianello et al. (159), which is generally consistent with the observations
of Gaston et al. (154) (TAS = 15 for both studies) for particles measuring 20–100 μm,
could be used to estimate exposure to the particles that are most likely to be trapped
in the upper airways during respiration and subsequently swallowed. Alternatively, the
study of Liao et al. (157) (TAS = 18) suggests an upper limit of 1583 ± 1181 MP/m3 for
particles measuring 5–30 μm, which could be used to estimate daily human exposure
via inhalation of approximately 3000 MP/day on the assumption of daily inhalation
of air of 15 m3. Consistent with the data on drinking-water (section 2.1), the variation
in the concentrations reported in air could be used for a probabilistic quantitative
assessment of human exposure (124).
2.3 Dermal exposure

Human exposure to NMP is largely dominated by ingestion
in food and beverages and by inhalation. Dermal exposure to
MP > 1 µm is limited by the barrier of the stratum corneum
(161), which comprises several layers of corneocyte cells
and individual packing of these cells in a “bricks-and-mortar”
structure of proteins and intercellular lipids. To reach the
systemic circulation, chemicals and other foreign agents,
such as particles, bacteria and viruses, must penetrate this
barrier.
26
2. Human exposure
Transdermal delivery of pharmaceuticals to optimize

permeation has been studied for centuries, including
investigation of the mechanisms by which chemicals and
particles with various physicochemical properties interact
with the stratum corneum (161–163). The strategies
include use of chemical permeation enhancers, biochemical
manipulations and physical disruption of the barrier function
(162, 163). Innovation in nanoscience has also included
evaluation of the conditions in which nanoparticles penetrate
the stratum corneum (162, 164, 165). Research is being
conducted on passive transcutaneous or lipid systems for
delivering NMP to avoid use of methods that disrupt or
compromise the integrity of the skin barrier. The methods are
based on delivery of antigens by passive diffusion through the
intact skin by establishing concentration gradients, increasing
the hydration of the skin by occlusion and diffusion into hair
follicles (165), which represent shunt pathways across the
stratum corneum that can be penetrated by particles (162,
166). Mahe et al. (166), for instance, observed penetration of
40- and 200-nm polystyrene nanoparticles into hair follicles,
and Vogt et al. (167) also observed penetration of 40-nm
nanoparticles. Penetration of particles into hair follicles may,
however, be influenced by particle size, those measuring
~ 600 nm being optimal, as this corresponds to the thickness
of the overlapping cuticular hair surface cells (167, 168).
Negatively charged, hydrophobic surface properties also
enhance follicular absorption (169).
While physiological variation in the thickness of the stratum
corneum and the density of hair follicles can influence the
results of experimental studies, three trends may be noted
(164).
• Experimental studies demonstrate consistently
that healthy human skin is a formidable barrier to
penetration of NP.
• Hair follicles are important collection sites for
nanoparticles, especially when skin is massaged or
flexed.
• The physicochemical properties of nanoparticles, such
as surface charge and hydrophobicity, significantly
influence permeation.
Kohli and Alpar (170) investigated permeation of negatively
charged 50- and 500-nm latex particles in a study of their
potential use for transcutaneous delivery of vaccines. They
observed no difference in the behaviour of the two sizes of
particle, < 0.1% of which permeated the stratum corneum.
Permeation was attributed to the repulsive force between
27
the negatively charged lipids in the skin and the particles.

Generally, limited permeation of nano-sized latex particles
is consistent with negligible exposure to nanoparticles in
cosmetics and personal care products across normal skin
(171). Indeed, one of the limiting factors for trans-follicular
vaccination is the relatively low follicular penetration of
nanoparticles, which is < 10% of the total applied amount
(168). Data on permeation of nanoparticles, however, vary,
and research is continuing on a dermal drug delivery system
based on “smart” nanoparticle systems that can release
active pharmaceutical ingredients at specific times and
locations by penetrating deep into hair follicles (169).
Observations on permeation of NMP across the stratum
corneum are, however, relatively limited, variable and
inconclusive (172), and no information is available on dermal
uptake of NMP during typical environmental exposure.
This information would be useful for overall assessment of
risks for human health. For instance, assessments should
be conducted of the safety of NMP in direct contact with
the skin, such as in application of nanotechnology in drug
delivery and the use of cosmetic and personal care products
(171). Research on dermal uptake of ambient NMP by
healthy intact and by damaged skin would also be useful.
Research on dermal exposure to NMP should build on the
decades of research on the application of nanotechnology in
the pharmaceutical sciences. Although MP in atmospheric
deposition and dust have been reported (3), the prevalence
of particles measuring < 10 μm is unknown, and methods
should be developed to characterize such exposure in
order to determine the importance of dermal exposure in
environmentally relevant scenarios.
2.4 Occurrence in food

The presence of plastic debris ingested by marine organisms was reported in a
number of studies conducted in the late 1960s and early 1970s (173–181) and
now includes observations of a broad range of MP in marine, freshwater and other
organisms (80, 182–192). MP may be ingested either directly from the water column
or sediment or indirectly by consumption of lower trophic prey that have recently
ingested MP (193). The evidence indicates that MP released into the environment can
enter the food chain, with implications for human exposure through consumption of
seafood and fish (7, 56, 85, 194–200).
In a review of studies of ingestion of plastic debris, including MPs, by > 800 species
and approximately 87 000 non-human organisms, the average concentration was
four particles per individual (201). Depending on the study, the concentration of
MP > 500 μm was measured in the stomach and intestines, with limited data on
28
2. Human exposure
concentrations in the tissues of fish consumed by humans. MP are therefore ingested

by non-human organisms, and laboratory studies indicate that egestion in faeces
represents an important elimination process (202). Thus, while MP are ingested at
all levels of biological organization, their detection may represent a snapshot in time
and space. Most (80%) of the organisms sampled did not contain MP at the time of
sampling, consistent with the estimate that 16.6% of organisms contained MP by
Hermsen et al. (28). Furthermore, the frequency of occurrence varied significantly,
ranging from 0 to 100%, depending on species, time of sampling, location and sample
size (28, 200, 201). The lack of standard methods for extracting and analysing MP in
organisms results in differences in reported data, such as on particle size, shape and
polymeric composition (200, 201). For instance, studies on the occurrence of fibres
in organisms began to appear only after 2011, when the release of synthetic textile
fibres was first reported (203). Since then, fibres and fragments have tended to be the
dominant shapes of MP observed; however, some studies included semi-synthetic
polymers in their analysis, while others excluded them. Natural, semi-synthetic and
synthetic fibres should be reported separately in the future, although natural textile
fibres tend to dominate the total number of fibres detected (204). Data on MP
< 100 μm in biological samples are limited, 5 μm being the lowest size reported so far.
Additional research is thus required to characterize and quantify the environmental
and biological fate of particles < 100 μm. MP have generally been studied only in the
gastrointestinal tract, and their presence in other biological tissues and organs is
usually either not studied or not reported (56, 200, 201, 205, 206).
Studies of commercial fish and seafood provide comparable results (56, 201,
205), with significant variation in the sample size and reporting of particles in the
gastrointestinal tract. As MPs are analysed only in the gastrointestinal tracts of fish,
which are usually removed before the fish are sold for human consumption, exposure
to MP in fish tissues is not certain and has been perceived to be negligible (10, 56,
206). Some laboratory studies have, however, found uptake of MP into tissues of fish
that are likely to be consumed by humans. This route of exposure should be better
characterized and quantified (83, 207–209).
Human exposure from consumption of seafood such as bivalves, crustaceans
and small fish, from which the gastrointestinal tract is not removed, may be more
relevant. Quantification of exposure from seafood depends, however, on the rate of
consumption and differences in the concentrations of MP. Estimated concentrations
are influenced by various factors, including the analytical methods, which may or
may not verify the polymeric composition of particles, and are directly influenced
by the inherent variation in environmental concentrations. Table 4 summarizes the
concentrations of MP reported in seafood and in other foods and beverages for
human consumption from studies identified in a search in PubMed with the keywords
“microplastic” AND “food” OR (“seafood” OR “human exposure” OR “beverages”).
The review was performed up to December 2021, and only those publications that
provided original concentrations were reviewed. Of the 87 studies identified, 58
reported concentrations in seafood or fish tissues, 18 reported concentrations in salt
products and four reported MP in beer; two reported MP in honey and milk and one on
MP in rice, sugar and nori seaweed. Each study was assessed for reliability according
to the 10 criteria of Hermsen et al. (28). The maximum possible TAS was 20, with
29
an average for all studies of 10.5. In the interests of transparency, details of all the
studies reviewed in this report are summarized in Table 4. No study received a non-
zero score on all criteria.
Seafood is the food product that has been studied the most often to date.
Normalization of the concentrations of MP in seafood per wet body weight (ww)
reported in Table 4 results in an average concentration of approximately 3 ± 4 MP/g.
In several studies, the concentrations in various food and beverages were combined
with estimates of per capita consumption to estimate dietary exposure (Table 5);
seafood represented the largest source of exposure. The importance of commercial
bivalves as a source of exposure to MP was first highlighted by Van Cauwenberghe
and Janssen (85), who estimated that the annual dietary exposure of European
consumers of molluscs was 1800 and 11 000 MP/year for the lowest and highest
consumers, respectively. Estimates based on meal portions of seafood by EFSA (10)
and the Food and Agriculture Organization of the United Nations (FAO) (83) and the
finding of 4 MP/g (ww) by Li et al. (273) indicate that an average portion of 225 g of
mussels would result in consumption of 900 MP per meal of mussels (Table 5). In a
study of cultural and regional differences in consumption of mussels, the exposure of
a consumer in the United Kingdom was estimated to be 123 particles/year, while that
of a consumer in Japan was estimated to be 56 210 particles/year (19, 160).
Table 5 summarizes estimates derived in studies of human exposure to MP based on
consumption of contaminated food. Table 4 shows significant variation in the quality
of the data reported and therefore in estimates of daily and annual human ingestion of
MP, so that it is difficult to compare the values with those in Table 5, and the difficulty
is compounded by differences in the particle sizes used to estimate consumption.
Caution should therefore be exercised in extrapolating the data for assessing the
implications of exposure to MP for human health.
While the values reported in Table 5 for seafood suggest that it may be an important
source of MP, only a limited range of types of food and beverages has been studied
so far, and they do not necessarily represent the major sources of daily caloric intake
by humans. For instance, the WHO Global Environment Monitoring System – Food
Contamination Monitoring and Assessment Programme (GEMS/Food) shows a
maximum per capita consumption of fish and seafood of 78 g/day in the cluster
G17 (consisting of Samoa and São Tome and Principe) and a minimum of 9 g/day in
cluster G1 (consisting of Afghanistan, Algeria, Azerbaijan, Gaza Strip and West Bank,
Iraq, Jordan, Libya, Mauritania, Mongolia, Morocco, Pakistan, Syrian Arab Republic,
Tunisia, Turkmenistan, Uzbekistan and Yemen). As illustrated in Fig. 4, cereals, grains,
fruits and vegetables (including roots) account for approximately 50% of the foods
ingested daily, although limited data are currently available on MP in these food
categories. The foods listed in Table 5 represent about 25% of the food categories
ingested daily. More data are required on food categories that better represent the
human diet for a more robust assessment of human exposure to MP.
30
2. Human exposure
Table 4. Reported numbers of microplastic or microplastic-like particles

and particle characteristics in studies of their presence in food
and beverages for human consumption
Lower Number of
ence Sample type (µm) (average) (average) Particle size (µm) shape polymer type score
91 Beverages 100 4.05 particles/L <2 Average fibre 98.4% as Not specified 8
(beer) particles/ length, 0.98 mm fibres
sample (0.1–5 mm)
210 Beverages 0.8 28 ± 9 MP/L 5 fibres/ Not specified Fibres and Not specified 6
(beer) blank fragments
211 Beverages 0.8 21.3 MP/L 16.7 Not specified Fibres and Not specified 7
(beer) particles/L fragments
212 Beverages 11 Maximum Laboratory 0.1–1 mm Fibres Polyamide, 8
(beer, cold values (MP/L): blank poly(ester-
tea, soft 6 (cold tea); 7 contained amide), PET and
drinks, (soft drinks); 6 no MP ABS
energy (energy drinks);
drinks) 28 (beer)
213 Beverages 11 6.5 ± 2.3 MP/L Laboratory 40% fibres < 0.5 Fibres Polyether 8
(milk) blank mm; 60% > 0.5 mm (97.5%), sulfone and
contained fragments polysulfone
no MP (2.5%)
214 Beverages 5 2040–10 040 44 ± 24 5–7 μm Fibrous Polyethylene, 17
(milk) MP/L MP/filter fragments polyester,
polytetraflu-
oroethylene,
polypropylene
89 Fish (dried) 149 1 particle/fish None > 149 μm Fragments Polypropylene 11
observed (85.7%), (47.2%),
in films (10%), polyethylene
procedural filaments (41.6%),
blanks (4.1%) polystyrene
(5.6%), PET
(2.8%), nylon-6
(2.8%)
215 Fish (dried) 20 0–1.92 ± 0.12 None 195–5780 μm Fibres Polyethylene, 10
MP/fish observed PET,
in polystyrene,
procedural PVC,
blanks polypropylene
216 Fish (tinned 149 1–3 particles/tin None 190–3800 μm Fragments Polypropylene 14
sardines and observed (46.6%), (33.3%), PET
sprat) in films (33.3%),
procedural (26.6%), polyethylene
blanks filaments (16.6%)
(26.6%)
217 Fish (tinned Not Not reported Not Not specified Not specified Nylon, 1,2-poly- 7
sardines, speci- specified butadiene,
tuna, fied ethylene vinyl
salmon) alcohol, wool
218 Fish 10 6.78 ± 2.7 MPs/ None 100–1000 Fibres Polyethylene, 15
(anchovy) fish observed polypropylene,
in polyamide,
procedural polyester,
blanks polystyrene
31
Lower Number of
219 Fish 20 No MP 0.095 MP/ Size of non-plastic Non-plastic Not specified 13
(seabass, sample fibres not reported fibres
muscle
tissue)
220 Fish 20 0.47 ± 0.84 MPs/ 0.25 ± 0.43 300–1000 Not specified Polyethylene 11
(various, fish MPs/fish and
fillets) Polypropylene
221 Fish Not 2.47 ± 2.99 to 0.40 ± 0.54 54–765 Fragments Polyethylene, 13
(various, speci- 0.47 ± 0.86 MP/ fibres/ polypropylene
fillets) fied fish blank
222 Fish 20 < LOD 0.4 MP/ Not specified Fibres Not specified 10
(various, blank
fillets)
223 Fish Not 0.12–0.51 MP/g Not 52.2% 100–250 μm Fragments Polyethylene, 11
(various, speci- specified polypropylene,
fillets) fied nylon
224 Fish Not Average, 5.7 ± Not > 100 μm Fibres Not specified 8
(various, speci- 1.7 and 18.5 ± specified
fillets) fied 4.6 particles/10
g fish muscle
225 Fish Not 0.74 ± 0.57 None 25–1000 μm Fragments Polyethylene, 15
(various, speci- MP/g observed polystyrene
fillets) fied in
procedural
blanks
226 Fish 8 Range, 4.23–9.3 0.57 ± 0.17 Average size 973 ± 97.9% fibres Cellophane 14
(various, MP/individual MP/blank 803 μm (skin) (33.5%),
skin) (skin) polypropylene
(15%),
polyethylene
(13%), nylon
(8%), polyester
(PET, 4.5%)
227 Fish and 5 2 MP/g (ww) 0.8 ± 0.131 5–25 μm 90% fibres PET, 11
seafood (maximum) items/ polyethylene
filter
228 Fish and 0.1 8.66E4 ± 2.43E4 Not 1.6–2.8 μm Not specified Not specified 7
seafood to 9.50E4 ± specified
6.64E4 MP/g
229 Fish and 30 0.26 ± 0.16 to Not 38.2–820 μm Fragments Polypropylene, 8
seafood 4.46 ± 3.72 specified polyethylene,
MP/g polystyrene,
PET
230 Honey 40 Fibres: 87 Not 40 μm to 9 mm Fibres and Not specified 4
± 73/500 g specified (fibres); 10–20 μm fragments
Fragments: 4 ± (fragments)
4/500 g
231 Honey 30 Not specified Not Fragments Mainly soot 10
specified and fibres or char; fibres
mainly cellulose
and PET
232 Nori 5 1.8 ± 0.7 MP/g 0.1 ± 0.2 0.1–4.97 mm; Fibres Polyester 12
(seaweed) (dry weight) MP/g (dry media, 1.13 mm (18.9%),
weight) rayon (6.6%),
polypropylene
(4%) polyamide
(2%), cellophane
(2%); cotton and
natural cellulose
fibres (61%)
32
2. Human exposure
Lower Number of
92 Rice Not 45–317 μg/g LOD Not specified Not specified Polyethylene, 14
speci- (dry weight) reported polypropylene,
fied (polyethylene); for PET
105 μg/g individual
(dry weight) polymers
(polypropylene);
17 μg/g (dry
weight) (PET)
90 Salt 149 2 particles/kg Collected Average size, 515 ± Fragments Polypropylene 12
(estimated) but results 171 μm and (40%),
not filaments polyethylene
specified (33%), PET
(6.7%),
polyisoprene/
polystyrene
(6.7%),
polyacrylonitrile
(10%),
polyamide-6
(3%)
91 Salt 100 212 particles/kg <2 Average fibre 99.3% as Not specified 8
particles/ length, 1.09 mm; fibres
sample range, 0.1–5 mm.
233 Salt 5 550–681 MP/ 4.4 ± 2.1 45 μm to 4.3 mm; Fragments PET, polyester, 7
kg (sea salt); particles/ particles < 200 μm and fibres polyethylene,
43–364 MP/ filter or 18 represented 55% polypropylene,
kg (lake salt); particles/ of total cellophane,
7–204 particles/ kg poly(1-butene)
kg (rock and well
salts)
234 Salt 5 127 ± 51.6 MP/ 6 30 μm to 3.5 mm Fibres PET (83.3%), 7
kg particles/ polyethylene
filter (3.3%),
polypropylene
(6.7%)
235 Salt 45 367 ± 154 to Not Not specified Fibres Nylon, 7
2133 ± 153 specified polyethylene
MP/kg
236 Salt 100 120–580 MP/kg None 100–500 μm Fibres Polyethylene 7
observed
in
procedural
blanks
237 Salt 50 1.68 ± 1.83 MP/ Not 3.3–4660 μm Fragments Polyvinyl 10
kg specified acetate,
polypropylene,
polyethylene
238 Salt 65 11–193 MP/kg None 65–2500 μm Fibres Polyethylene 16
observed
in
procedural
blanks
239 Salt Not 2 ± 1 to 72 ± 40 Not 55–2000 μm Fibres Polyethylene, 7
speci- MP/kg specified polypropylene,
fied polyester
240 Salt 20 8–102 MP/kg 2.03 ± 1.01 20–5000 μm Fibres Polyethylene, 10
MP/salt polyurethane,
type polypropylene
33
Lower Number of
241 Salt 10 1570–31 680 Not 10–4628 μm Fragments Polypropylene, 7
MP/kg specified and fibres polyamide, PET,
PVC
242 Salt 100 672 ± 2560 MP/ LOD, 100–5000 μm Fragments Polyethylene, 16
kg (median = 82) 0.72 MP/ polypropylene,
kg (PET PET
fibres)
243 Salt 390 6.7–53.3 MP/kg Not 390–9360 μm Not specified Polyethylene, 1
specified polyvinyl
acetate,
polystyrene
244 Salt 5 < 700 MP/kg Not 5–3800 μm Fragments Cellophane, 6
specified and fibres polystyrene,
polyamide,
polyarylether
245 Salt Not 275 ± 25 to 1832 Not 20–5000 μm Fragments Polyethylene, 5
speci- ± 40 MP/kg specified polypropylene,
fied PET, nylon and
polystyrene
246 Salt 500 56 ± 49 to 103 ± Not 500–2000 μm Fragments PET, polyamide, 9
39 MP/kg specified and fibres polyethylene,
polystyrene
247 Salt 1 140.2 MP/kg None 89.7–1474.9 μm Fragments Polypropylene, 9
observed polyethylene,
in PET
procedural
blanks
58 Salt 20 2395 MP/kg None 63–100 Fragments Polypropylene, 12
observed polyethylene
in
procedural
blanks
248 Seafood 1.2 0.0–5.47 5.8 ± 2.2 Not specified Fibres (90%) Not specified 16
(clams) particles/g (ww) particles/
filter
249 Seafood 20 0.06-5.17 MP/g Not 62.3% < 500 μm Fibres Rayon, polyester 12
(clams) (ww) specified
250 Seafood 10 23 ± 20 MP/clam None 39% < 500 μm Fibres PVC, 12
(clams) observed polyethylene,
in polypropylene,
procedural polyester
blanks
251 Seafood 100 3 MP/individual Not Not specified Fibres Polyethylene 4
(clams, specified
oysters)
252 Seafood 0.5 Not specified Not 0.5–5 mm Fibres and Not specified 3
(crab) mm specified fragments
253 Seafood Not 34–178 items/ Not Not specified Fibres Not specified 11
(mussels) speci- mussel specified
fied
254 Seafood Not 6.2 ± 7.2 items/g None 750 μm to 6 mm Fibres Not specified 8
(mussels) speci- (ww) observed
fied in
procedural
blanks
160 Seafood Not Average, 0.09 6.5 ± 0.95 Fibre length, 0.2–2 Fibres Polyester, PET 13
(mussels) speci- ± 0.03 MP/g MP/blank mm; thickness,
fied (ww) to 3.0 ± 0.9 1–5 μm
MP/g (ww)
34
2. Human exposure
Lower Number of
255 Seafood 5 2.2 items/g 0.67 ± 0.82 33 μm to 4.7 mm 65% fibres PET, polyester, 16
(mussels) (ww); 4 items/ items/ fibres) cellophane
mussel filter
256 Seafood 5 0.13 ± 0.14 1 fibre/ 77% < 50 μm Fibres and 9
(mussels) MP/g (ww) blank fragments
257 Seafood 10–20 3.5 fibres/10 g LODs of 200–1500 μm Fibres 8
(mussels) μm (ww) 2.3, 4.7
and 1.5
fibres/
sample
for black,
blue and
red fibres,
respec-
tively
258 Seafood 1.2 0.9 ± 0.2 MP/g < 10% of 100–500 μm 77.8% Polyethylene, 17
(mussels) (ww) total MP fragments; PET, polystyrene
detected in 22.2% fibres
blanks
259 Seafood 5 1–5.4 MP/g 0.4 ± 0.5 250 μm to 1 Fibres (80%) PET (74%), 12
(mussels) (ww) items/ mm (48–76% of polyethylene,
filter particles) PVC,
polyethylene,
rayon
260 Seafood 25 1.05–4.4 MP/g 19 fibres/ Median length, 1.2 Fibres and Polyamide, PET 13
(mussels) (ww) tape strip mm fragments
261 Seafood 1.2 37 MP/g (dry Reported Median length, Fibres Not specified 9
(mussels) weight) as minimal 200 μm
262 Seafood 0.7 Not specified Not 20 μm to 5 mm Fragments Not specified 14
(mussels) specified
263 Seafood 5 0.7–2.9 MP/g 0.67 ± 0.75 5–250 μm Fibres Polyester, 11
(mussels) (ww) items/ polypropylene,
filter polyethylene,
rayon, cotton
264 Seafood 1.6 0.76 ± 0.40 MP/ Not 32.6% 100–500 μm Fibres and Polyethylene, 15
(mussels) individual; 0.15 ± specified fragments polypropylene,
0.06 MP/g (ww) polystyrene,
ABS, PET,
styrene–
butadiene
rubber
copolymer
265 Seafood 8 0.86 ± 0.82 0.67 ± 0.58 890 μm (average) Fibres Cellophane, PET 9
(mussels) MPs/g (ww) items/
filter
266 Seafood 500 0.04 MP/g (ww) Not 72% > 500 μm Fragments PET 7
(mussels) specified and
filaments
267 Seafood 500 0.87 ± 0.55 to Not 39% 500–1000 μm Fibres Polyethylene 15
(mussels) 10.02 ± 4.15 specified
MPs/mussel
268 Seafood Not 8.72 ± 5.30 MP/ Not > 100 μm Fibres Not specified 11
(mussels) speci- mussel specified
fied
269 Seafood 53 1.53 ± 2.04 Not Not specified Fragments Ethylene/ 11
(mussels) MPs/g (ww) specified propylene
copolymer
35
Lower Number of
270 Seafood 30 0.08 to 8.6 MP/g 1.33 ± 0.58 41.7–4679 μm Fragments Polypropylene, 16
(mussels) (ww) Polyethylene,
PET
271 Freshwater 2.85 ± 1.27 None 44.8% < 100 μm Fragments Not specified 13
Mussels MP/g observed
in
procedural
blanks
85 Seafood 5 Average, 0.24 ± None 5–25 μm Fragments Not specified 8
(mussels, 0.07 to 0.35 ± observed
oysters) 0.05 particles/g in
(ww) procedural
blanks
272 Seafood 20 0.61 ± 0.56 1.2 ± 0.8 50–100 μm (52%); Fragments Polypropylene 18
(mussels, (mussels) particles/ 20–50 μm (37%); > (80%) and
oysters) and 2.1 ± 1.7 filter, none 100 μm (11%) polyethylene
(oysters) MP/ verified as
individual or 0.2 plastic
± 0.2 MP/g (ww)
273 Seafood 5 Average, 2.1– 0.5 ± 0.55 5 μm to 5 mm; Fibres and Polyethylene, 10
(mussels, 10.5 items/g items/ 33–84% < 250 μm fragments PET and
scallops, (ww) filter polyamide
clams)
274 Seafood 20 Average, 0.15 ± LOD, 0.24/ Fibre lengths, 43 Fragments > 80% 17
(mussels, 0.2 MP/g (ww) sample μm to 4.7mm; (76%); fibres polyethylene,
scallops, or 0.97 ± 0.74 (polyeth- MP < 300 μm (24%) polypropylene,
clams, MP/individual ylene, represented 65% polystyrene,
oysters) polypropyl- of total polyester and
ene, poly- expanded
styrene) polystyrene
and 0.51/
sample
(polyes-
ter and
polyeth-
ylene vinyl
acetate)
275 Seafood 5 0.37– 0.57 MP/g Not Not specified Fibres and Polyamide, PET 12
(oysters) (ww) specified fragments
276 Seafood 25 Average, 0.2–20 None 37–58% were 58% fibres, Polyethylene, 10
(oysters, particles/g (ww) observed 10–25 μm 26% PET and nylon
clams, and 3.5–17.7 in fragments,
snails) particles/ procedural 14% films
individual blanks and 2%
pellets
277 Seafood 10 4.0 ± 2.1 MP/g Not 10–428 μm Fibres Ethylene vinyl 11
(oysters, (oysters); 3.2 specified alcohol
mussels, ± 1.8 MP/g
clams) (mussels); 0.7
± 0.3 MP/g
(clams)
278 Seafood 5 0–5 MP/ Fibres Not specified Not specified Not specified 12
(Clams, individual excluded
mussels, (mussels); 0 MP from anal-
crabs) (crab soft tissue ysis due to
and clams) contami-
nation
36
2. Human exposure
Lower Number of
279 Seafood 74 0.31 ± 0.10 Not 74–2000 μm Fibres Cellulose, 10
(Prawn, MP/g (oysters/ specified polyamide,
oysters, clams); 0.25 acrylonitrile,
clams) ± 0.08 MP/g polyethylene,
(prawns) polypropylene,
PET
280 Seafood 5 3.24 ± 1.02 None 53% < 100 μm Fragments PET (69.4%) 11
(oysters) MPs/g (ww) observed
in
procedural
blanks
281 Seafood Not 64 MP/g (ww) Not 30–5000 μm Fibres Not specified 8
(oysters) speci- specified
fied
282 Seafood Not 0.07 ± 0.04 Limited 75% > 500 μm Fibres PET, 14
(oysters) speci- MP/g (ww) to a few polyacrylonitrile,
fied polyamide rayon
fibres
198 Seafood 500 0.6 ± 0.9 MP/ Not 0.1–4.5 mm Fibres Not specified 8
(Pacific oyster specified
oyster)
194 Seafood Not 1.5 MP/g (ww) Not > 100 μm Fibres Not specified 11
(prawns) speci- specified
fied
283 Seafood 10–20 0.68 ± 0.55 Reported 200–1000 μm 96.5% fibres Not specified 11
(prawns) MP/g (ww) or as < LOD
1.23 ± 0.99 MP/ (not
prawn assessed)
284 Seafood 100 6.78 ± 2.8 MPs/ Not 100–1000 μm Fibres Polyethylene, 12
(prawns) prawn specified polypropylene,
polyamide,
nylon, polyester,
PET
285 Seafood 100 1.02 MP/g (ww) Not 87% > 100 μm Fibres PET, 8
(prawns) specified polypropylene,
polystyrene
230 Sugar 40 Fibres: 217 Not 40 μm to 9 mm Fibres and Not specified 4
± 123/500 g specified (fibres); 10–20 μm fragments
Fragments: 32 ± (fragments)
7/500 g
ww, per wet body weight

Additional information on the total assessment score (TAS) for each study available upon request. The
maximum TAS was 20.
37
Table 5. Estimated daily and annual per capita ingestion of microplastic

particles
Daily per
Por- capita
tion consump- Particle Per capita Per capita
Refer- size tion (g/ concentration ingestion ingestion Particle
ence Sample (g) Country or region day) (MP/g)a (MP/day) (MP/year) size (μm)
10 Mussels 225 France 4 900b 25
268 Mussels 18.16 g/ 70.82 > 100
year
274 Mussels Republic of Korea 0.67 0.12 0.08 29 4–300
263 Mussels 100 United Kingdom 100 5–250
160 Mussels United Kingdom 0.23 1.5 0.3 123 0.2–2 mm
160 Mussels Belgium, France, 8.75 1.5 12.7 4620 0.2–2 mm
Spain
270 Mussels 836–5672 21-458 176–10 380 > 40
g/year
228 Mussels 0.21 237 1.6–2.8
277 Mussels 3109.3 g/ 8084.1
year
254 Mussels (cooked) 225 Italy 1395 > 750
249 Clams 1088.64
274 Manila clams Republic of Korea 1.25 0.35 0.44 155 43–300
274 Oysters Republic of Korea 0.84 0.07 0.06 21 43–300
274 Scallops Republic of Korea 0.25 0.08 0.02 7 4–300
283 Prawns Belgium 1.4 1.92 0.3 100 > 200
19 Seafood 37.82 1.48 55 20 430
227 Fish and seafood United Kingdomc 20.74 0.77 16 5828 >5
227 Fish and seafood United Kingdomd 57.2 0.77 44 16 076 >5
224 Fish muscle Persian Gulf 45 1.85 80 29 200 > 100
(bartail flathead;
Platycephalus
indicus)
228 Fish sea (bream) Mediterranean Sea 22.5 25 500
223 Fish India 43 65
225 Fish Islamic Republic 43 174 MP/kg
of Iran bw per year
216 Canned fish Global 0.25 0.03 0.008 2.7
89 Dried fish Bangladesh 1.01 0.7 0.7 246 > 149
215 Dried fish Sri Lanka 3700 g/ 851–1147
year
19 Salt 5 0.11 0.55 200 > 30
91 Salt USA 5 0.21 1.05 380 > 100
234 Salt Spain 5 0.28 1.4 510 > 30
233 Salt China 5 0.68 3.4 1 241 > 45
90 Salt Malaysia 10 0.002 0.02 7.3 500
238 Salt Sri Lanka 8.3 158
239 Salt India 10.98 48–216
240 Salt Turkey 14.8– 63.7–302.4
18.01
58 Salt Republic of Korea 10.06 12 000 > 20
38
2. Human exposure
Daily per
Por- capita
tion consump- Particle Per capita Per capita
Refer- size tion (g/ concentration ingestion ingestion Particle
ence Sample (g) Country or region day) (MP/g)a (MP/day) (MP/year) size (μm)
242 Salt Global 10.06 0–117 0–42 600 > 100
19 Sugar 66.81 0.44 30 10 730 > 10
19 Honey 2 0.1 0.2 73 > 10
92 Rice 100 Australia 3.7 mg ± 1.4
(unwashed
rice); 2.8 mg
± 0.3 (washed
rice); 13.3
mg ± 2.5
(microwaved)
213 Milk Mexico 0.36 L/ 6.5 ± 2.3 MP/L 2.4 858 10–500
day
91 Beer 0.35L USA 4.05 MP/L 1.42 520 > 100
19 Alcohol 0.04 32.27 MP/L 1.3 470 > 100
19 Bottled water 0.44 L/ 94.37 MP/L 40 15 155 >5
day
19 Tap water 3.26 L/ 4.23 MP/L 13.8 5 030 > 100
day
1 Drinking-water 2L/day 10.4 MP/L 20.8 7 592 150
19 Inhalation 170 62 050 > 50
a
Maximum or average value
b
Concentration per meal, not per day
c
Based on per capita consumption derived by the United Kingdom Department for Environment, Food and
Rural Affairs (227)
d
Based on per capita consumption derived by FAO (227)
39
Fig. 4 Dietary consumption from 16 food categories derived from all

17 GEMS/Food clusters
Source: https://www.who.int/teams/nutrition-and-food-safety/databases/global-
environment-monitoring-system-food-contamination, accessed 16 May 2020.
Inset summarizes the average percentage of each food category in total per capita dietary
consumption of foods in all 17 GEMS/Food clusters
To address the lack of data on different food categories, standard methods should be
developed for consistent, robust assessment of exposure. Table 4 summarizes the
TAS of studies on MP in food and beverages. The only matrix for which substantial
resources have been invested in developing an analytical method is seafood, although
quality assurance and control are limited, with variation in sample sizes, reporting of
concentrations in individual organisms, whether and how many samples were pooled,
different methods for extracting MP from tissues, particularly with respect to tissue
digestion, variation in processing of procedural blanks, filter substrates and pore
sizes and different approaches to categorizing particles as MP, including both visual
inspection and analytical verification of polymer composition.
Food categories other than seafood have been analysed by methods for which the
performance has not been verified, with low TAS for studies of MP in sugar, honey,
salt, beer and other beverages. Most of the reports did not provide details of the
efficiency of recovery of MP of various shapes, sizes and polymer composition, which
could result in underestimation of the concentrations. Furthermore, the samples of
food items tended to be small (i.e., < 25 individual items) purchased on a single date
from a limited number of suppliers, usually without production lot numbers or dates,
raising concern about their use for extrapolating concentrations of MP in a specific
food category. To assess dietary exposure, both the sample sizes and the temporal
40
2. Human exposure
and spatial trends should be substantially increased for a robust statistical analysis of
variations in concentrations of MP.
The availability of standard analytical methods, consistent with the elements
summarized in Box 2, is fundamental for assessing human dietary exposure to
NMP and particularly quantification and characterization of particles < 10 μm. As
summarized by the EFSA Panel on Contaminants in the Food Chain (10) and by FAO
(83), MP > 150 μm are unlikely to be absorbed, and uptake of smaller MP is expected
to be < 0.3%. As discussed in section 3, particle size and shape influence systemic
uptake, distribution and elimination. Particles > 10 μm that are inhaled, for instance,
are generally understood to be trapped in the upper airways and subsequently
swallowed. NP < 0.1 μm in the gastrointestinal tract can potentially be taken up
systemically (7% estimated by FAO (83)), and the probability of egestion increases
with size. Quantification of human exposure to NMP < 10 μm is essential, as the
effects on human health increase with decreasing particle size. Table 5 indicates,
however, that most studies have addressed exposure to MP > 10 μm, which are
probably excreted directly (1, 10, 83).
Research should also be conducted on the sources and characteristics of NMP in
food and beverages in order to introduce effective, efficient measures to reduce
exposure. In the characterization and quantification of MP in seafood, for instance,
it appears to be assumed that the environment is the main source of contamination,
whereby filter feeders, such as mussels, ingest and accumulate MP from
contaminated seawater and sediment (85, 254). Other studies suggest contamination
during processing of food and beverages and from packaging. Li et al. (263), for
instance, reported significantly greater contamination of processed than of live
farmed mussels, suggesting that MP were introduced during de-shelling and cleaning
rather than by ingestion and accumulation from the environment. Karami et al. (216)
suggested that tinned fish are contaminated during preparation and packaging.
Kutralam-Muniasamy et al. (213), Kosuth et al. (91) and Iñiguez et al. (234) suggested
focusing on steps in the processing of milk, beer and salt at which contamination
with MP might occur. Liebezeit and Liebezeit (210) suggested that MP and other
anthropogenic debris are introduced into German beer during manufacture, whereas
Lachenmeir et al. (211) proposed that a microfiltration step to remove yeast cells
from processed beer would be sufficient to also remove MP. They further suggested
that observations of MP in beverages such as beer are an artefact due to poor quality
assurance and quality control, with contamination occurring by deposition of MP onto
samples in the laboratory.
Deposition of MP from the air has been suggested as a further source of dietary
exposure. In a comparison of direct exposure to MP from the consumption of
mussels and exposure in household dust, Catarino et al. (160) estimated that
exposure to MP due to deposition was more than two orders of magnitude greater
than that from ingestion of contaminated mussels. As foods can be contaminated
by deposition from air and/or in processing and packaging, additional research is
necessary to characterize and quantify relevant sources of contamination. A number
of studies have recently addressed the relative importance of food packaging (see
for instance 92–94, 286–295). The observations provide valuable preliminary insight
into the role of plastic packaging; for example, heating plastic containers appears to
41
increase the release of NMP (92, 93, 287, 289–291, 293), with pH also possibly playing
a role (94). Concern has also been expressed about the lack of standardized methods
required for robust assessments (292, 296, 297). More research on the role of plastic
packaging should be conducted for quantitative assessment.
Methods for determining the polymeric composition of particles should be included
in future studies. Of the 87 studies in Table 4 on MP in food, 27 did not provide
confirmation that the particles were plastic, and 29 provided limited verification. Thus,
only about one third of studies that reported the occurrence of “microplastic” provided
satisfactory analytical verification. Widespread reporting of MP in food in the absence
of verification is an obvious problem for estimating human exposure. For instance,
in their analysis of MP in honey, Liebezeit and Liebezeit (230) reported an average
concentration of 87 ± 73 items/500 g of honey and suggested that the honey was
contaminated by plastic fibres attached to pollen, during processing of the honey or
from packaging. They did not, however, provide analytical verification that the fibres
were plastic, a concern raised by Mühlschlegel et al. (231), who included analytical
verification and reported limited contamination of honey by MP.
Estimates of dietary exposure to MP will require significant advances in:
• development of standard analytical methods appropriate for characterizing and
quantifying various foods and beverages, with application for particles < 10 μm;
• targeted sampling to identify sources of contamination throughout manufacture,
processing and packaging of foods and beverages; and
• characterization of the contamination of food and beverages by deposition
during preparation and consumption.
Given the limited data on exposure to contamination in important food categories,
illustrated in Fig. 4, an intelligent sampling strategy is necessary to clarify the
differences among various sources of contamination and for robust quantification of
human exposure. Contamination of cereals, grains, fruits and vegetables with NMP
should be characterized, as some research suggests contamination from agricultural
soils (298, 299). Agricultural practices include use of a variety of plastic products (e.g.,
plastic film for mulching, vinyl tunnels, fertilizer bags) that may introduce NMP into
agricultural soils and into products (300–302). Application of biosolids to soil is also
an important source of MP in the terrestrial environment (300, 303).
Although concern has been expressed here about the quality of the reporting of MP
in food and beverages, recent research by Mohamed Nor et al. (124) suggests a
probabilistic approach to estimating human exposure. For instance, they estimated
that the total daily median MP mass intake from nine media (fish, mollusc, crustacean,
tap water, bottled water, salt, beer, milk and air) was 0.2 (0.0001–7500) μg/child per
day and 0.6 (0.0003–17000) μg/adult per day (124). Comparison with the estimate by
the World Wildlife Fund that potential exposure to MP is 700 mg/person per day (304)
suggests that the latter estimate represents the 99th percentile intake of an average
person.
Current approaches to assessing human exposure to MP from food are all based on
combining data on dietary absorption rate with data on the amount of MP in food
components. In several studies, exposure was assessed by deterministic estimation
of the total intake from all dietary components, (19, 124, 304, 305). A major limitation
42
2. Human exposure
of these studies is use of databases with different definitions of MP and use of

different analytical techniques. As a result, the data are inconsistent, including
different ranges of MP size, obviating confident quantification of human exposure.
In addition, exposure estimates based on average exposure rates do not reflect
the actual distribution of global MP intake rates. These issues could be resolved in
probabilistic approaches, such as that of Mohamed Nor et al. (124). In this approach,
lifetime exposure is assessed for intake of eight food types that represent 20% of the
human diet by weight. Intestinal absorption, biliary excretion and plastic-associated
chemical exposure were also estimated (section 5.2), in both MP number and mass
concentrations. The probabilistic model provided estimates of MP concentrations in
body tissues, gut and stool, the last of which could be compared with empirical data.
To ensure the comparability of data on microplastic abundance, re-scaling was done
to account for the different size ranges and types of particles targeted by the different
methods used (306–308). The simulated microplastic concentrations in stool were
15% of those in the available empirical data, which would appear to be consistent, as
only 20% of the human diet was taken into account. This demonstrates the usefulness
of probabilistic approaches for estimating human exposure.
2.5 Summary and recommendations

Human exposure to MP has been characterized in air,
drinking-water, food and beverages, and research should be
conducted to differentiate the sources of contamination.
Given the ubiquitous exposure to MP, standard methods
should be used to characterize and quantify NMP of the sizes
considered most relevant to human health, i.e., < 10 μm. Most
studies to date have addressed MP > 10 μm, and the results
are incomplete for assessing risk, as the implications for
human health increase with decreasing particle size.
Assessment of exposure by inhalation will require methods
for quantifying and characterizing NMP associated with the
respirable fraction of atmospheric particulates. Methods for
sampling particles with aerodynamic diameters associated
with PM2.5 and PM10 could be used. Data on atmospheric
deposition of MP indicate the potential sources and
processes that influence environmental fate and transport
and could be used to characterize the fraction of MP
associated with ingestion of dust.
Studies of the presence of MP indicate that exposure occurs
via inhalation and by ingestion of drinking-water, food and
beverages, although the relatively small number of samples,
food categories and populations limits a comprehensive
assessment of exposure for the implications for human
health (4, 13, 141, 309, 310). Concern has also been raised
about the quality of studies. Standard methods are required,
with investigations of laboratory contamination, and which
43
ensure consistency in the analysis of particles within the

relevant size range, such as 1–5000 μm, to allow comparison
of studies and to estimate human exposure. A key challenge
is characterization and quantification of NMP measuring
< 10 μm. Significant analytical challenges include extracting,
isolating and verifying the polymeric composition of particles
of such sizes, and it is also likely that the background
contamination increases with decreasing particle size. A
robust quality assurance and quality control protocol is
therefore necessary.
Key messages
• Human exposure to NMP is ubiquitous and occurs by all routes.

• Information on exposure from air, drinking-water, food and
beverages is limited. Data on the characteristics of NMP and their
quantification in each of these media are necessary, with better
understanding of their sources.
44
3. OBSERVATIONS FROM
EPIDEMIOLOGY
Within the Global Burden of Disease programme (311), it was estimated that, in
2015, 4.2 million people had died prematurely due to exposure to airborne PM.
The components of PM that represent the greatest risk to human health, however,
are poorly understood, although a contribution of NMP cannot be excluded (111).
At present, exposure estimates and routine measurements for environmental
epidemiological studies are lacking. Such studies should include exposure to NMP
and to other types of particles (e.g., from combustion sources) and gases, with
consideration of confounding factors. Effects due to long-term exposure can be
measured only from exposure estimates over periods from years to decades, which
would exclude retrospective studies. Effects of short-term exposure may be foreseen
in the near future, with observation of spatial and temporal variation in the exposure
of a population for whom sufficient information on health is available. Until then,
the best information is from occupational epidemiology, in which subgroups of the
general population who often experience exposure well above ambient levels are
studied. As such studies usually do not include vulnerable people, the findings cannot
be extrapolated to the general population; however, the findings can provide insight
into pathology related to NMP exposure. Most regulatory jurisdictions define limits of
exposure to particulates in the workplace, such as those in the United Kingdom of
10 mg/m3 as an 8-h time-weighted average for inhalable dust and 4 mg/m3 for
respirable dust (76). Like the guidelines set for PM, these exposure limits are for dust
in general and are not specific for NMP. Studies of controlled exposure of humans to
synthetic fibres and particles to simulate occupational exposure are extremely rare
because of ethical considerations (312, 313). Studies have, however, been conducted
on people exposed occupationally to mixtures of various fibrous and non-fibrous
plastic particles at high concentrations over extended periods (314).
One of the earliest documented outbreaks of disease related to exposure to synthetic
fibres by inhalation was reported in 1975 among workers in the textile (nylon,
polyester, polyolefin, acrylic) industry (reviewed in 315), in which workers showed
symptoms of allergic alveolitis. Outbreaks of occupational interstitial lung disease
have since been reported in the manufacture of nylon “flock” (316, 317), which are
short fibres produced for making velvet-like textiles and upholstery. When they are
produced in a rotary mill, for instance, a substantial amount of respirable nylon dust
is generated, leading to an average exposure concentration of 2.2 mg/m3 (314).
Otherwise healthy, often young workers in this industry develop respiratory symptoms,
including chest pain, shortness of breath and cough (318). The bronchoalveolar
lavage fluid of such workers shows an abnormal cellular profile. Nonspecific
interstitial pneumonia is established, with accumulation of lymphocytes, lymphocytic
inflammation of the bronchioles and in some cases proliferation of lymphocytes
in alveolar tissues. Flock workers’ lung is, however, a rare disease; for example, it
has been diagnosed in 24 workers in North America (317, 319–322). Although the
condition is debilitating, it is usually reversible, and the respiratory symptoms stabilize
45
and ultimately improve after removal from exposure (317, 320), which also suggest
that the NMP are cleared from the lungs over time, unlike some other particles or
fibres. In some cases, however, the condition evolves to fibrosis and respiratory failure
(317, 320, 321).
Similar pathological presentation and symptoms have been reported in workers
exposed to other synthetic flock, such as polyethylene (323), polypropylene (324) and
rayon (325), raising concern that exposure to high levels of non-specific polymeric
organic fibres increases the risk of interstitial lung disease. Long-term occupational
exposure to respirable cotton (and flax and hemp) fibres is also associated with lung
disease, respiratory symptoms and loss of pulmonary function, and asthma and
chronic obstructive pulmonary disease have been documented (326). Although the
adverse effects of synthetic fibres appear to be due mainly to their physicochemical
properties and high concentrations, the adverse effects triggered by occupational
exposure to respirable cotton fibres are suggested to be due to an endotoxin secreted
by Gram-negative microbes on the surface of cotton fibres (326).
It has been suggested that exposure in the nylon flocking industry increases the
risk of lung cancer. In a retrospective study of 162 workers in a nylon flocking plant,
the risk was three times higher than that of controls (327). Moreover, workers in
a polyester and polyamide fibre factory in France were found to be at statistically
significantly greater risk of death from various cancers (n = 79; relative risk, 1.42;
95% confidence interval, 1.06 ; 1.89 for high exposure; and n = 105; 1.38, 1.05; 1.81 for
previous exposure to polymer dust), irrespective of the level or duration of exposure
(328). These findings should be corroborated in a larger cohort study, with adjustment
for confounding by individual smoking histories and other lifestyle factors to allow
extrapolation to other exposure scenarios. A study of female textile workers in China,
however, found no association between exposure to synthetic fibres and lung cancer
risk (329). It is therefore difficult to draw conclusions about the carcinogenic risk of
exposure to microplastics.
Histopathological analyses of lung biopsy samples from synthetic textile workers
exposed to various polymers showed not only interstitial fibrosis but also
granulomatous lesions containing foreign bodies considered to be acrylic, polyester
and nylon dust (315).
Exposure to PVC particles has also been linked to disease, predominately in
occupational settings. Inhalation of airborne PVC dust has been associated with
interstitial lung disease, as determined by chest radiographic abnormalities (330, 331).
In a cross-sectional study of 818 PVC workers, forced expiratory volume in 1 s and
forced vital capacity were inversely related to exposure to dust, after adjustment for
age, height and smoking. The response was observed mainly in cigarette smokers,
suggesting an interaction between smoking and PVC dust. The authors concluded
that, while an average dust index (age × mg/m3) of 12.9 caused a partial decrease
in lung function (a loss of 53 mL over 20 years, in addition to losses due to age and
smoking), workers exposed to higher levels might suffer an important loss of lung
function (331). Exposure for 60 days to total dust (of which PVC particles < 1 μm were
the predominant component) at a concentration of 0.3–42 mg/m3 (median, 2 mg/m3)
resulted in severe dyspnoea, a decrease in transfer factor (diffusing capacity), profuse
46
3. Observations from epidemiology
accumulation of macrophages with some haemorrhage and increased numbers of

elastic and collagenous fibres in one individual (332). The patient had had minimal
exposure to low concentrations (0–3 mg/m3) of airborne vinyl chloride monomer, for
which pulmonary effects usually occur after exposure to higher concentrations over
a longer period, and had not been exposed to asbestos or other known fibrogenic
agents. The presence of PVC particles in the lung indicated that they were probably
responsible.
Epidemiological studies have also been conducted on the potential relation between
occupational exposure to plastic dusts and cancer. Demers et al. (333) reported an
association between current exposure to plastic dusts and lymphocytopenia; however,
later sequential analysis was not predictive, implying no clear association. Earlier
studies identified a risk of cancer among women working in the plastics industry, and
later studies investigated possible cause–effect relations (334–337). Sorahan and
Nichols (336) found no relation between occupational exposure to amine catalysts,
non-flammable and flammable solvents, polyurethane dust, latex, rubber, curled hair
or coir fibre, feathers or foam handling and the risk for lung cancer among female
workers. Limitations to the studies have, however, been identified, including lack
of data on smoking, uncertain exposure estimates, estimates only for exposure by
inhalation and, in some instances, limited cohort size.

Numerous epidemiological studies have been conducted
among workers exposed to various NMP in the plastics
and textiles industries. Studies of occupational exposure
to particulates do not, however, reflect the exposure of the
general population, and caution is warranted in extrapolating
results for various types and concentrations of particles
associated with occupational activities to indoor and
outdoor environments. Data obtained from occupational
epidemiology may, however, be helpful in identifying hazards
and pathological adverse effects after long-term, elevated
exposures to specific NMP. There is some evidence of
specific lung pathology in occupational settings. There is
inadequate evidence of the carcinogenic risk of exposure to
NMP.
Key messages
• Evidence in the literature that inhalation or oral uptake of NMP can

affect the gastrointestinal tract or other organs apart from the lung
is limited and of inadequate quality.
• Better estimates are required of exposure of the general population
to NMP and co-pollutants by inhalation and in the diet.
47
4. DOSIMETRY AND
BIOKINETICS
Assessment of the risks posed by xenobiotics in studies in experimental animals
usually requires extrapolation from high to low doses and extrapolation of data from
animal species to humans. Extrapolation may require clarification of the exposure
pathway. For example, if exposure to airborne particles results in deposition only in
the upper respiratory tract, most, if not all, of the particles will be swallowed, resulting
in oral exposure. The toxicity of those particles can thus be tested directly by oral
exposure of experimental animals.
This section summarizes studies on how exposure, dose and biokinetics influence
the uptake of NMP by the body, their distribution among organs and their clearance.
Considerable research has been conducted on the biokinetics of particulates,
including NMP of specific shape, size and polymeric composition. An important
consideration is extrapolation of observations on well-defined particles to the
concentrations and properties of NMP in the environment. This section provides
summaries of studies on various types of particles. While caution should be exercised
in extrapolating laboratory results to the heterogeneous mixture of NMP in the
environment, the data can inform future studies and provide a perspective of the
implications for human health.
4.1 Dosimetry: extrapolation from external to

internal exposure
4.1.1 Dosimetry: inhalation
Exposure-dose–response relationships are the basic model
for assessing the effects of stressors, activity patterns and
exposure on various biological end-points in assessing
risks to human health. As toxicity is related to the dose
of a substance, it is important to determine whether an
external exposure can provide an internal dose capable of
causing an adverse effect. This information is also required
in extrapolating results obtained in vitro or in experimental
animals to humans. The challenge is to simulate human
physiology realistically, to evaluate the kinetics of particles in
experimental animals and cells and to use biokinetic models
to extrapolate measured doses in cells to humans in vivo
(129, 338–341). Fig. 5 provides an example of extrapolation
of concentrations and doses from rats to humans.
The dose of particles required to elicit an adverse effect at
the most critical biological target depends on factors that
influence their deposition and clearance. “Particle deposition”
refers to deposition onto tissues in the respiratory tract, which
49
Fig. 5 Attributes of NMP to be considered in assessing both exposure

and hazard
Source: reference 342 (https://creativecommons.org/licenses/)
is influenced by aerodynamic behaviour (inhalation) and

thermodynamic or electrostatic interactions (340). Although
various metrics can be used (e.g., mass, number, surface
area, volume), deposited mass is most frequently used to
describe the relationship with a toxicological response (308,
340). Clearance of inhaled deposited particulates depends
on the initial site of deposition and the physicochemical
properties of the particles, which determine the relative
importance of specific biokinetics and biodynamics.
Considerable research has been conducted on the dosimetry
of inhaled particles, resulting in clear understanding of the
biokinetics of the deposition and clearance of insoluble
particles such as carbon black, coal, diesel soot, talc and
titanium dioxide (341–343). Particle size, shape, density,
surface properties (i.e., surface charge, area and chemistry)
and the distribution of those properties influence the
deposition of all types of particle, whereas clearance is
related not only to the properties of the particle but also to its
physiological location in vivo (342, 344).
50
4. Dosimetry and biokinetics
The epithelial surfaces of the respiratory tract are the

largest interface between humans and the environment. For
instance, the respiratory tract is estimated to cover > 100 m2,
while the gastrointestinal tract, the next most important
interface, covers 30–40 m2 (345, 346). The epithelium of the
respiratory tract is constantly exposed to a heterogeneous
mixture of particles suspended in inspired air (346, 347), and
the fractions of respired particulates deposited in various
regions of the respiratory tract have been investigated
and modelled for a range of particle sizes and ventilatory
patterns (129, 131, 348). Many of these aspects are included
in particle dosimetry models, such as the multiple-path
particle dosimetry model (349). The latter can be used to
model particle deposition and clearance in humans (age-
specific) and experimental animals (rat, mouse, rabbit, pig
and monkey) for extrapolation of results from one species to
another (Fig. 5).
4.1.2 Particle inhalation models

The mechanisms of particle deposition in the respiratory tract include settling
or sedimentation, in which the density of particles is an important factor; inertial
impaction, in which changes in the direction of flow can determine whether particles
deposit on surfaces; and diffusion, which is influenced by Brownian forces between
gaseous molecules and particles, which cause particles to collide and deposit onto
surfaces. Secondary processes of deposition include interception (typical for fibres)
and electrostatic interactions between predominately positively charged particles and
the negatively charged surfaces of the respiratory tract (347).
Fig. 6 illustrates the relations among particle sizes deposited in different regions
of the respiratory tract. Particles ≤ 10 μm (mass median aerodynamic diameter,
MMAD, the diameter above and below which 50% of particles in the aerodynamic
size distribution lie, according to mass) deposit mainly in the nasopharyngeal region.
Smaller particles, including those measuring nanometres, are more likely to deposit in
the alveolar pulmonary regions due to diffusion (Fig. 6) (347).
In view of the heterogeneity of size, shape, density and surface charge of NMP,
models may be useful for screening and identifying the most hazardous combinations
of properties. The properties relevant for particle inhalation include a neutral or
negatively charged surface and a relatively narrow distribution of densities, centred
on about 1 g/cm3. It is likely, for instance, that positively charged NMP interact with
the larger fraction of naturally occurring negatively charged particles, causing them
to aggregate into a larger, heterogeneous mixture of particles, whereas neutral NMP
can be inhaled directly or undergo weathering to a predominately negatively charged
surface. The material density of the most commonly encountered plastic materials
(Table 1), such as polyethylene, polypropylene, nylon and polyester, is usually
0.85–2.3 g/cm3, although the effective density in a matrix or an agglomerate may be
lower. Particles of the same geometric diameter but with different (effective) densities
51
Fig. 6 Main regions of particle deposition in the human respiratory

system and modelled deposition of a 1-g/cm3 spherical particle
in relation to the diameter of the particle
Source: Adapted from references 347 and 349

D, diameter
have a different deposition pattern: lower densities result in smaller aerodynamic

diameters.
All the mechanisms of deposition are strongly influenced by the pattern of breathing,
with significant differences between breathing patterns at rest and during exercise in
individuals of different ages and sex (347). Fig. 6 shows important differences in the
deposition pattern of particles. For particles > 2 μm MMAD, there is a stronger (but not
exclusive) likelihood of deposition in the upper respiratory tract (head, nasopharynx)
and upper airways (tracheobronchial), while particles measuring 0.01–1 μm are
deposited deeper in the lung (pulmonary, alveoli, gas exchange area). The deposition
of particles in relation to their size distribution can be compared with the typical
particle size distribution of PM in an urban area.
The dose of particles in the respiratory system therefore depends on their
(aerodynamic) size, micrometre-sized particles being deposited mainly in the upper
respiratory tract and particles > 10 μm MMAD being less likely to move beyond the
nose and mouth. Most deposited particles with a diameter > 2.5 μm MMAD will be
transferred to the digestive system, as they tend to be swallowed, whereas smaller
particles are more likely to interact with tissues of the alveoli, where they can be
engulfed by alveolar macrophages and transported to the mucociliary escalator
or become senescent. The main mechanism for deposition of nanoparticles is
diffusion, indicating that slow air speed such as in alveolar spaces and near the
olfactory epithelium will result in a higher likelihood of deposition. Typically, there is
little deposition when diffusion and impaction are less dominant, which is the case
52
for particles of 0.2–1.0 μm MMAD. Monitoring of the physicochemical properties,

particle size distribution, density and surface properties of NMP in air would add
important data for models of particulate inhalation and support characterization and
quantification of human inhalation exposure.
Particle shape should also be considered, as experimental data indicate that fibres
reach the lower airways more efficiently than spheres (350–352).
4.1.3 Particulate inhalation: experimental studies

Inhalation of particulates, including NMP, by experimental
animals in vivo has been studied for several decades. For
instance, Stemmer et al. (353) administered a single dose
of 5 mg polyurethane dust to weanling and 9-month or older
rats to assess the pulmonary inflammatory response. The
dose was based on the total amount of dust estimated to
be inhaled by rats exposed for 8 h per day for 30 days; the
MMAD was < 5 μm for 52% of the particles and < 10 μm for
93.5%. Exposure was by intratracheal instillation, a commonly
used method for introducing materials into the lungs of
test animals because of its simplicity, relatively low cost,
delivery of a well-defined dose, delivery to the rodent lung of
particles that are not respirable by rodents but are relevant to
humans (such as long fibres) and for practicality and safety
(354). This method of exposure does not, however, represent
physiological exposure, limiting extrapolation for evaluating
pulmonary toxicity. As summarized by Driscoll et al. (354) in
a critical review, there are inconsistencies between dosing by
instillation and by inhalation, and data based on instillation
should be interpreted cautiously for evaluating respiratory
toxicity; limiting assumptions should be stated and guidelines
followed carefully. These caveats should be kept in mind in
interpreting the toxicological data summarized in section 5.
More recent studies have involved approaches for better
understanding the adverse effects of exposure dose and
specific stressors. For instance, Xu et al. (355) compared the
adverse effects of spherical PVC particles measuring
0.2–2.0 μm and crystalline fragments of silica measuring
0.5–3.0 μm. Crystalline silica particles were used as a positive
control because of their strong association with silicosis, lung
cancer and autoimmune diseases, the crystalline structure
of the particles accounting for their pathogenicity (356). Xu
et al. (355) administered the particles to male Wistar rats
by intratracheal instillation at 10 or 50 mg/kg body weight
(bw). The differences in the adverse effects of PVC and silica
particles were attributed to differences in their biokinetics,
PVC particles being cleared more efficiently than those of
53
crystalline silica. The mechanism of the slow clearance and

biopersistence of silica particles is poorly understood; however,
it would appear that the surface-dependent properties of silica
particles, such as the reactivity of silanol groups and changes
in surface area that increase surface reactivity due to particle
fracture, influence clearance rates (356–358).
Translocation of NMP from the respiratory tract epithelial
surfaces has been proposed, with concern about the ultimate
fate and potential accumulation of NMP in the body. The
size of insoluble particles influences their translocation into
interstitial spaces, decreasing size being associated with
greater potential translocation. Ferin et al. (359) observed
that particles measuring 20–30 nm penetrated the interstitial
space more easily than those of 200–500 nm. In a study of
polystyrene nanoparticles (average diameter, 56.4 nm and
202 nm), Chen et al. (360) gave male Sprague-Dawley rats
0.6 mg of radioiodinated polystyrene particles by intratracheal
instillation and found that only a small fraction passed rapidly
into the systemic circulation but that translocation was
markedly increased after infusion of lipopolysaccharides to
induce pulmonary inflammation. The use of radiolabelled
particles helps to characterize and quantify the fate and
transport of particles and thus strengthens understanding
of the mechanisms of translocation, including physiological
processes (e.g., pulmonary inflammation) in relation to the
size and shape of particles.
The shape of NMP is important (361). Although atmospheric
monitoring indicates exposure to a wide range of fragments
and fibres (136), most experimental studies of the adverse
effects of NMP in animals have been conducted with
spherical particles; a few studies involved exposure to
respirable fibres. Warheit et al. (362) exposed male rats
(Crl:CD(SD)IGS BR) to aerosols of nylon-66 respirable fibres at
0, 4, 15 or 57 fibres/cm3 by nose-only inhalation for
6 h/day for 20 exposures. The nylon-66 fibres were described
as white, 18-denier, trilobal, chopped and ground fibres > 99%
pure with a mean length of 9.8 µm and a diameter of 1.6 μm.
The results showed rapid lung clearance over 12 months,
with a no-observed-effect level of 57 fibres/cm3, the highest
concentration tested.
In general, complementary use of dosimetry models and
careful study design, including controls, are required to
understand the mechanisms of effects on human health
of inhalation of NMP (see section 5). NMP that are
representative of environmentally relevant exposure should
be characterized and quantified.
54
4.1.4 Dosimetry: ingestion

The physicochemical properties of particles, such as size, shape, surface area and
chemistry, are usually measured in a dry state and then used to compare a biological
response to an oral dose (363, 364). In the same way as for inhalation, the relation
between particle properties and an adverse effect may, however, be influenced by
interactions between particles and between particles and the physiological fluids or
food suspension used to deliver the dose (364–368). Dispersion of particles in the
dosing medium is therefore an important factor to consider; homogeneous dispersion
allows robust interpretation of results (363, 365, 368). The fate of particles in the
dosing medium influences various dose metrics, including the actual delivered mass
and particle number (364, 366, 369). Delivery of particle stressors to cell systems
in vitro or to animals in vivo to evaluate their fate after oral ingestion may vary, and
standardized methods are necessary for comparisons and evaluations, not only
between in-vivo test systems but also for quantitative in vitro to in vivo extrapolation
(QIVIVE).
4.1.5 Dosimetry: in vitro

Cell culture systems, which are simple models of complex
biological systems, have provided valuable insight into the
biological, physiological and pathological responses of cells
exposed to chemical and physical stressors; however, these
systems have limitations, including uniform distribution
of a dose, particularly for particles (363, 364, 368, 370,
371). Administered particle doses are usually reported as
an initial mass or number concentration, which does not
represent the dose actually delivered to cells over time
(371). Actual exposure is to particles that come into direct
contact with cells and can trigger an adverse effect, which
is not necessarily equivalent to the nominal concentration
of particles in the medium (370). The mass, surface area
or number of particles delivered to receptor sites on the
cell surface or the corresponding area under the time–
concentration curve reflect the dose at the site of action
better than the nominal media concentration (338, 371). The
delivered dose is also determined by interactions between the
physicochemical properties of the particles, the properties
of the medium in which the particles are suspended and the
duration of the test. Particles are subjected to dispersive
forces in the test medium, which can result in formation of
larger agglomerates. This in turn influences the effective
density of the particles, further affecting deposition kinetics.
Standard methods are thus required for dispersion and
dosing of NMP in in-vitro systems to ensure adequate
analysis of adverse effects (369, 371–373). The properties of
the cell systems themselves should also be considered, as
55
the presence of a sticky mucus layer, such as in co-cultures

of epithelial and goblet cells, or the presence of macrophages
can influence dose deposition and uptake kinetics, whereas
monoculture cell systems are devoid of mucus (364).
Interpretation of data from in-vitro models often depends
on the nature of the exposure and the complexity of the
models (367). Most systems involve submerged cultures. To
ensure quality and for extrapolation to an equivalent human
dose, several critical parameters should be reported to allow
computational modelling of the dose and dose rate, essential
for extrapolating data to in-vivo models:
• the biologically effective dose (369–371), i.e., the
number or mass of particles in direct contact with a
certain number or surface of cells (cm2);
• the dose rate and duration of exposure (371);
• the size (and size distribution) of the primary particles
and the agglomerates or aggregates (16, 371, 374);
• sample preparation (e.g., use of proteins to disperse
the plastics stably and sonication, which may generate
reactive oxygen species) (16, 374); and
• the presence of contaminants, such as chemical
toxicants or endotoxins (16, 374).
Other requirements may be added, according to the study
hypothesis. For example, a study of the relevance of size for
uptake into cells would require at least two sizes of plastic of
the same composition and appropriate controls for assessing
the influence of surface properties such as area, charge and
reactivity (374). A special case is use of “air–liquid interface”
exposure systems, in which cells are exposed to NMP via an
aerosol under static or dynamic flow conditions, resulting
in a more realistic but also more complicated system to
assess the deposited dose. Some exposure systems include
a microbalance in parallel to the exposed cell surface, which
gives a fair indication of the dose delivered over an exposure
period.
An appropriate dose range is particularly important for in-vitro
studies, because cell monocultures or co-cultures differ from
cells in an organ and from physiologically relevant oral and
inhalation exposure (364). There is currently no standardized
method for determining the dose to be administered in vitro
that is equivalent to published estimates of human exposure.
56
Table 6. Criteria for evaluating the reliability and quality of in-vivo and
in-vitro studies on the effects and biokinetics of nano- and
microplastic particles
Criterion Comment
Human exposure See section 2. Exposure must be characterized to identify
characterization environmentally relevant concentrations and properties of NMP.
Specifically, size, shape, polymer composition and surface properties
should be quantified and relevant exposure pathways (inhalation, oral,
dermal) characterized.
Characterization • particle size distribution
of NMP • agglomeration state
biokinetics • particle shape
• chemical composition
• particle surface area
• surface chemistry and charge
Characterization Important to increase the quality of data on the relation between dose
of NMP after and an adverse effect, as the physicochemical properties and dose may
administration change during administration in either in-vivo or in-vitro test systems.
Method of Details of how particles are introduced into an in-vivo or an in-vitro test
administration system, i.e., type of inhalation exposure, such as intratracheal or nose-
only, solvent or delivery vehicle used, method of aerosolization, sample
volume, cell surface
Duration of Times of observation and, ideally, particle properties at each time
exposure
Use of negative Necessary to evaluate the performance of the study by demonstrating
and positive that the adverse effects associated with particles, such as crystalline
controls silica (positive control), are consistent with other observations and that
negative controls, such as a solvent or vehicle, provide an appropriate
baseline for interpreting adverse effects.
4.1.6 Dosimetry: summary and implications

This section addresses the dosing of NMP for assessing the adverse effects of
their inhalation and ingestion and characterizing and quantifying the biokinetics.
Several methods for the dosimetry of particles are summarized, with their challenges
and implications. Dosimetry is critical to the quality of a study. A testing strategy
will contribute to hazard identification and risk assessment only if the dose, route
of exposure and particles are fully characterized (374, 375). The principles are
summarized in Table 6. Many are included in tools for evaluating toxicity screening
methods, such as the modified version of the ToxRTool (374) and the NMP toxicity
study assessment tool (16), which was used to evaluate the quality of studies cited in
section 5.
4.2 Biokinetics
Once a foreign substance enters the body by inhalation
or ingestion, it may or may not cross the biological
barriers and be distributed in the body. Some substances
accumulate in lipid-rich tissues, for instance, while others
are readily eliminated via the urine or bile, transported to the
gastrointestinal tract and excreted in faeces. The process that
57
influences physiological uptake, distribution and elimination

throughout the body is known as biokinetics. Metabolism,
another important biokinetics mechanism, is not expected to
influence the physiological fate of NMP (83, 86, 376).
4.2.1 Inhalation
After deposition of NMP in the respiratory system (Fig. 6), they are cleared by various
mechanisms that effectively eliminate inhaled particles. These include mechanical
mechanisms, such as sneezing, mucociliary clearance, phagocytosis by alveolar
macrophages and lymphatic transport (312).
The basic clearance processes are solubility, macrophage function, mucociliary
transport, cellular endocytosis, intercellular sieving and lymph and capillary blood
flow. The site of deposition in the respiratory tract and the physicochemical properties
of particles, such as size, shape and surface reactivity, influence the clearance
mechanism (129, 131, 347).
Numerous studies have been conducted to characterize the clearance of insoluble
particles deposited on surfaces in the lower respiratory tract (see, e.g., 342, 377).
Clearance is generally understood to occur in two phases: an initial phase with a half-
life of 3–12 h in the tracheobronchial region and a second, alveolar phase that may
last several months or longer (347). In experiments with whole-body exposure of rats
to PVC particles at 8.3 mg/m3 for 25 h/week for 7 months, the average mass of PVC
retained in the lungs 1 month after cessation of exposure was 2 mg/lung (378).
Marked differences in clearance kinetics at high dose levels have been observed
between rodents and humans, and caution should be taken in extrapolating
observations from experimental animals.
Mucocillary clearance of particles depends on the mobility of mucus, a viscoelastic
secretion that protects the mucosa from dehydration and provides a medium for
inhaled particles. As a result of constant ciliary action, the mucus flows and is
eventually cleared from the airway and transferred to the digestive system, from which
particles are egested (341, 347, 379). Inhaled particles that reach the alveolar region
of the lung can be transferred into the interstitium. Their presence at the epithelial
surface can also stimulate chemotactic signals that attract alveolar macrophages
to the site of particle deposition, where phagocytosis by the macrophages initiates
particle clearance from the alveolar region. Alveolar macrophages, however, have
a finite lifespan and decay, releasing any undissolved particles for phagocytosis by
another alveolar macrophage. Particles that cross the alveolar epithelium into the
interstitium may encounter interstitial macrophages, initiating a process similar to
that for alveolar macrophages. A fraction of particles can also be transferred from the
interstitium to lymph nodes. Modelling of this process is informative for assessing
risks for human health of inhaled particulates (341, 379). Another fraction of particles
may be retained in the interstitium.
Kevlar para-aramid fibrils have been shown to be biodegradable in the lung, as the
recovered fibres appeared to be “shorter” than the original fibres (380). The Kevlar
fibres were reported to have a half-life in the lung of 30 days. The mean length of
58
Kevlar fibres recovered from digested lung tissue decreased from 12.5 μm to 7.5 μm
over 6 months after exposure, and the mean fibre diameter decreased from 0.33 μm
to 0.23 μm. Warheit et al. (362) assessed mucociliary clearance of nylon fibres in
a 4-week nose-only study in rats and measured the recovery and dimensions of
fibres several times after exposure. They reported a rapid decrease in the number
of recovered nylon fibres at 3, 6 and 12 months after exposure (from an exposure
concentration of 57 fibres/cm3). In this study, the fibre lengths did not change up
to 180 days after exposure, indicating that biodegradability does not affect lung
clearance of nylon fibres. Rapid lung clearance of inhaled nylon fibres was, however,
reported, with an estimated half-time clearance of inhaled fibres of about 2 months at
the high exposure and 1 month at the medium exposure.
Recently, MP were detected in lung tissue collected during routine coroner autopsies
of 20 non-smoking adults aged 48–94 years (381). A total of 31 synthetic polymer
particles and fibres were observed in 65% of individuals, dominated by fragments with
a mean particle size of 3.92 ± 1.96 μm. Polyethylene and polypropylene were the main
plastic polymers detected, and 16% of particles were identified as cotton. Although
inhalation is understood to be the most likely exposure route for particles observed
in the lung, Amato-Lourenço et al. (381) did not rule out the possibility that some
particles may reach the lungs by systemic translocation.
4.2.2 Ingestion
Particles that are ingested are considered to be available
systemically only when they are absorbed by the intestinal
epithelium, pass through the liver and are distributed via the
bloodstream throughout the body. A number of physiological
barriers significantly limit the absorption and systemic
bioavailability of particles from the gastrointestinal tract,
although local absorption may occur. As discussed in the
previous WHO report (1), microplastics > 150 μm ingested
from drinking-water are expected to pass through the
gastrointestinal tract without being absorbed.
A fundamentally important physiological barrier in the
gastrointestinal tract is mucus, a selectively permeable
hydrogel that acts as a physical barrier to particle diffusion
across the epithelial tissues. The main structural component
of the mucus layer is mucin, a highly glycosylated protein with
oligosaccharide side-chains that include terminal sialic acid
and sulfate residues, resulting in a net negative charge (382).
The average pore size of the mesh-like structure formed
by the interactions of mucins is 10–500 nm. The mucus
layer significantly impedes the diffusion of small particles
by interaction filtering (i.e., electronic and hydrophobic
interactions) and can fully block the penetration of larger
particles by both steric (i.e., size) and interaction filtering. The
rate of passage of particles along the gastrointestinal tract
59
is usually effective in ensuring that most ingested particles

are excreted. Szentkuti (383) studied the rate of diffusion of
carboxylated fluorescent latex NP of various sizes across the
mucus layer to the enterocyte surface and found that
14-nm particles passed through the mucus layer within 2 min
and 415-nm particles within 30 min; however, micrometre-
size particles did not diffuse through the mucus. Although
the author observed permeation of both 14- and 415-nm
particles, none of the particles was endocytosed by the
enterocytes but appeared to move in the opposite direction
with mucus. Therefore, while particle size is an important
factor in permeation of mucus, the accumulation of particles
on the enterocyte surface is insufficient to trigger phagocytic
events, and the mechanism probably does not represent a
significant translocation pathway.
O’Hagan (384) identified several physiological sites
of particle uptake according to their size, by ordinary
enterocytes, intestinal macrophages, the epithelium of
Peyer’s patches and the villus tips. The main function of
the epithelial cells (enterocytes) is to absorb and transport
nutrients for systemic distribution (385); however, they can
also endocytose particles in the nanometre range, such as
partially digested dietary ferritin (~12 nm in diameter) from
meat and plant foods (386). In an early study, Sanders and
Ashworth (387) suggested that ordinary enterocytes are
responsible for the uptake of 200-nm polystyrene particles in
rats, although Jani et al. (388) proposed an upper size limit
of 100 nm for uptake by enterocytes. Garrett et al. (389) also
observed uptake of polymeric nanoparticles by enterocytes
in the mouse gut in vivo, using 30–50-nm ammonium
palmitoyl glycol chitosan particles. The particles were then
transported to the liver through the circulatory system and
were recirculated through the bile to the small intestine and
excreted in the faeces. According to Yoo et al. (390), the
upper particle size limit for endocytosis is about 500 nm.
It has been proposed that particles > 500 nm are absorbed
by intestinal macrophages (390), as observed for 1-µm
polystyrene microplastics in dogs and rats (391). Uptake of
particles > 1 μm is, however, currently considered to occur
most frequently across specialized “microfold cells” in
Peyer’s patches, which are domed regions that form gastro-
associated lymphoid tissue (384, 392). There are, however,
important differences in the rate of uptake by enterocytes and
by Peyer’s patches in the large and small intestine (393–395).
Microfold cells sample lumenal microparticles such as
viruses and bacteria (1–10 µm) and transport them to
60
the underlying lamina propria by a process known as

“transcytosis” (396). Most studies on transcytosis have
been conducted with latex microplastics. As latex has no
pathogenic activity, observation of their absorption suggests
that microfold cells are “particle agnostic” and are possibly
influenced by the physicochemical properties of particles,
such as size and surface charge (396).
Translocation and absorption of microparticles < 10 μm
across microfold cells has been observed after active
phagocytic transport of various inert materials from the gut
lumen, with the particles migrating to the blood via mesentery
nodes and the thoracic lymph duct (397). Microfold cells
therefore not only represent a conduit through which
particles can permeate Peyer’s patches but also play a key
role in the body’s immune surveillance, actively sampling the
contents of the gastrointestinal tract for potentially harmful
particulate antigens. This implies that particles transported
by microfold cells first enter the lymphatic system and that
this immunological barrier facilitates clearance of foreign
substances by immune cells (394, 398).
The rate of uptake across Peyer’s patches depends on the
size of particles. After a single oral dose (12.5 mg/kg) of
polystyrene microspheres to female Sprague–Dawley rats,
uptake was rapid for those measuring 50 nm, moderate
for 500-nm and slow for 1-μm microspheres (388). After
the Peyer’s patches, particles are translocated towards
the mesentery node via mesentery blood and lymph
vessels (10 days after daily dosing) (399), and 50-nm
fluorescent polystyrene microspheres were found in the
mesentery networks, their concentration peaking 12 h after
administration; the levels of 500-nm microspheres peaked
between 12 and 18 h after administration. It is hypothesized
that particles enter the mesentery network and vessels
via phagocytes and open lymphatic tubules in the Peyer’s
patches. After 18 h, particles were present in liver and spleen
(388). While 1-μm spheres were present at lower levels, they
persisted in Peyer’s patches, the mesentery network and
nodes for 36 h, and most of the 50- and 500-nm spheres were
transported to the liver and spleen within 24 h.
Another mechanism of translocation of microplastics of
the size of those in foods and beverages is villous uptake.
Evidence of this mechanism was obtained over a century
ago in studies of various microparticle types. A phenomenon
referred to as “persorption” has been described, which is
passive absorption of microparticles measuring 5–150 μm
from the intestinal lumen through gaps in the mucosa that
61
may result from mechanical kneading of single-layered

intestinal epithelium resulting from cell shedding, particularly
at desquamation zones of the villi (383, 400–402). Volkheimer
(400, 401) conducted many experiments in animals and in
humans and observed that starch particles up to 130 µm in
diameter could be detected in blood after ingestion. Following
persorption, particles are immediately removed from the
intestinal wall into the lumen of blood and lymph vessels
and are finally excreted; 12 h after ingestion of a starch
suspension, for instance, only a few starch granules were
observed in peripheral blood, and negligible numbers were
present after 24 h (402). The phenomenon of persorption
and its influence on uptake of MP should be studied further
with standardized methods and materials. Generally,
however, absorption of particles > 10 μm is considered
to be negligible, whereas absorption of particles < 10 μm
increases with decreasing size. In a review in 2012, Carr et
al. (403) concluded that the mechanism of uptake of small
microparticles (1–5 μm) is almost entirely villous, at least
after a single exposure.
Overall intestinal absorption of MP is reported to be low. Carr
et al. (403) conducted experiments in various rodent species
and observed that only 0.04–0.3% of latex MP measuring
2 µm was absorbed. Pathological tissue, e.g., tissue from
patients with inflammatory bowel disease, may transport
more particles than healthy tissue (0.45% as compared to
0.2%), as shown by Schmidt et al. (404) in human colon tissue
in vitro in an Ussing chamber. The reason was suggested
to be greater permeability of gut tissue in inflammatory
bowel syndrome than in healthy tissue. Stock et al. (405)
investigated the uptake of 1-, 4- and 10-µm polystyrene MP
in vitro in the Caco-2 monolayer, the microfold cell model
with specialized cells and the mucus model, and in vivo in
HOTT mice. Consistent with the observations of Carr et al.
(403), negligible numbers of particles were found in cells
of the jejunum and duodenum of mice in vivo (405), and no
particles were found in any other organ. Caco-2 cells took up
more 1- and 4-μm particles (≤ 0.8% and 3.8% of total particle
recovery, respectively) in vitro, whereas few 10-µm particles
were found (0.07% recovery). The authors suggested that
fewer 1-μm particles were absorbed because they were taken
up only by phagocytosis, while the 4-μm particles may have
been absorbed by both phagocytosis and micropinocytosis,
although the 1-μm particles may also have lower settling
rates, reducing their availability for absorption. Furthermore,
the surface chemistry of the two particle sizes differed, and
greater absorption of 4-μm particles might have been due
62
to the presence of a sulfate group. Both the 1-μm and the

4-μm particles were recovered at significantly higher rates
in co-cultures than in the Caco-2 monoculture. The authors
concluded that only a minor fraction of small MPs enters the
intestinal wall (405).
The potential for NMP to cross the intestinal barrier is thus
understood to occur in a size-dependent manner whereby the
uptake and transport of particles measuring up to 5–10 μm
into intestinal cells is possible and intracellular uptake of
larger particles is unlikely, because of incompatibility with
the size of intestinal epithelial cells (about 10 μm) (310).
Consistent with observations by EFSA (3), FAO (66) and the
previous WHO report (1), NP may thus be absorbed, although
the rate of absorption appears to decrease with increasing
particle size, becoming negligible for particles > 150 μm.
Caution should, however, be exercised in extrapolating from
the results of the available studies, which are limited to
a homogeneous group of polymers and sizes that do not
necessarily represent the heterogeneous mixture of NMP
encountered in the environment.
Polystyrene NP, for instance, have been used in numerous
studies of toxicity in mammals in vivo and in vitro (reviewed
in section 5). The oral bioavailability of 50-nm and 100-nm
neutral and positively and negatively charged particles was
investigated by Walczak et al. (406) in vitro. Size was a major
determinant of translocation of NP, up to 7.8% of 50-nm NP
and 0.8% of 100-nm NP being translocated. Surface charge
and chemistry were also found to influence translocation.
A study in rats in vivo by the same group indicated ≤ 1.7%
particle uptake in kidney, heart, stomach wall and small
intestine wall, which was less than that in the in-vitro study
(407). Sinnecker et al. (408) added fluorescent latex NP of
various sizes to isolated perfused rat small intestine. They
found no particles in the vascular or lymphatic systems but
a significant amount in lumen samples. The results indicate
that intestinal tissue provides a sink function for NP. Most
particles were detected in the mucus lining and did not
permeate the epithelium. The authors concluded that a
healthy small intestine provides an effective barrier against
NP uptake and is strongly influenced by the health of the
mucus film layer.
The rates of uptake and translocation depend not only on
size but also on intestinal location and time. Oral exposure
of male Sprague-Dawley rats to a single dose of polystyrene
microspheres (1.6 µm; 1.65 × 109 microspheres per animal)
resulted in maximum concentrations (representing 1.4%
63
of the initial dose) in the proximal segment of the Peyer’s

patches 0.5 h after administration. The percentage of
microspheres taken up by Peyer’s patches decreased with
both location (i.e., towards the distal regions) and time (409).
In a study with polymethyl methacrylate nanoparticles, 95%
of the dose administered to rats was eliminated within 48 h
(410).
In a study of eight individuals, a median of 20 MPs/10 g of
stool was observed, and the particles detected ranged in size
from 50 to 500 μm (411). More recently, Zhang et al. (412)
reported detection of MP in the stool of 96% of participants
(n=26) at concentrations ranging from 1 to 36 MP/g and
sizes from 20 to 800 μm. These two studies provide only
preliminary assessments of human exposure to MP and
subsequent elimination and recommend that more robust
mass balance studies be conducted to characterize and
quantify excretion as an effective elimination pathway and
possibly for use in biomonitoring to improve understanding of
human exposure. Additional research should be conducted on
NMP and particularly on their retention and translocation.
4.3 Biokinetics: summary and recommendations

To better understand exposure to and the main mechanisms of biodistribution of
NMP from experimental results in animals, consideration should be given to use of
experimental data, such as from in-vitro bioassays, in biokinetic models, such as
physiologically based pharmacokinetic (PBPK) models and their use to complement
understanding of how NMP are retained, cleared, translocated and distributed in
the body (367, 413, 414). PBPK models that have been developed for engineered
nanomaterials could be used to estimate the retention, clearance and translocation of
NMP. Use of existing models would indicate similarities in behaviour and also, perhaps
more importantly, differences. By understanding the physicochemical properties of
NMP and the physiological processes that might influence NMP and nanomaterials
differently, research can be directed to assessing the implications for human health.
Whether NMP are inhaled or ingested, their biokinetics is strongly influenced by
their size, shape, density and surface chemistry. The experimental data summarized
above suggest that caution should be exercised in extrapolating from the limited
data available, but they are sufficient to conclude that MP > 150 µm are unlikely to
be absorbed and that absorption increases with decreasing particle size, via oral
exposure. MP < 1 µm, which include the nano-sized fraction, are most likely to be
absorbed, but characterization and quantification of uptake are limited.
Research to strengthen exposure assessment would complement dosimetry and
biokinetics. Specifically, quantitative data on the (rate of) translocation of NMP and
on size distribution, shape, polymer composition and surface chemistry in air, food
and beverages, including drinking-water, are necessary to determine the properties
64
of NMP that are most relevant for use in studies of biokinetics and effects. This will
require different measurement techniques according to the media in which the NMP
are dispersed.
• Physiological mechanisms for the uptake, distribution and

elimination of MP minimize tissue exposure. The probability of
Key messages
uptake into the body increases with decreasing particle size.

• There is insufficient information to assess biodistribution (uptake,
retention, clearance, rate of translocation), including the likelihood
that NMP will cross biological barriers after deposition on the
epithelium tissue or after reaching the circulation
• Dosimetry models are available for extrapolation of results on
particle inhalation obtained in experimental animals to humans, but
they have not been evaluated or validated for NMP.
• Data on the biokinetics of NMP obtained in models in vitro cannot
currently be extrapolated to the situation in vivo.
65
5. TOXICOLOGICAL EFFECTS
The sections above show that MP are ubiquitous in the environment but that the data
on exposure in the diet and by inhalation are insufficient for quantitative assessment
of exposure. Furthermore, there is concern about the use of non-standard methods
for generating the data and the fact that monitoring has been mainly of particles
measuring > 10 μm (section 2). There is thus great uncertainty about human exposure
to biologically relevant NMP measuring < 10 μm. We recommend that research be
conducted on the adverse effects of NMP in studies which account for their dosimetry
and characterization, with quantification of the properties of particles, such as size,
shape, surface properties and polymer composition, and biokinetics (section 4).
Although uncertainty about human exposure to NMP is a significant barrier to
assessing risks to human health, several studies have reported adverse effects both
in vivo and in vitro, and occupational epidemiological data are available (11, 12, 26,
195). Better understanding of the toxicological effects of NMP will require studies
of the relations between particle properties and their toxicity. The physicochemical
properties of particles, such as their size, shape and surface chemistry, are
understood to contribute to some toxicological end-points. Thus, the toxicity of a
fragment or fibre can be attributed to interaction of the particle with tissues, the
effect of a chemical or biological contaminant on the particle, including desorption
of chemicals or pathogens on the surface, and the complex interaction of several
factors (415). This section summarizes studies on the toxicological effects of plastic
particles and fibres, particularly those due to physical interaction with particles.
The studies were identified in a literature review in PubMed with the keywords
“microplastic”, “microplastics” AND “toxicity” and were supplemented by studies
referenced in published reviews on the toxicity of NMP. Additional references on
synthetic fibres were obtained from the reference list in a report by the Health and
Safety Executive in the United Kingdom (416) on the hazards and risks of fibres,
supplemented by a search with the keywords “synthetic fibre” AND “toxicity” OR
“health” in PubMed up to December 2021. As noted in the introduction, although
every effort has been made to ensure that the literature reviewed and evaluated for
this report is as comprehensive as possible, it is not possible to guarantee that every
study has been captured in this rapidly emerging field.
All studies identified were evaluated with a recently published NMP toxicity study
assessment tool, the purpose of which is to screen and prioritize studies for risk
assessment according to their reliability, which is scored on a number of criteria
of quality assurance and quality control (QA/QC) (16), summarized in Fig. 7, which
include consideration of:
• identification of the test substance,
• characterization of the test system,
• description of the study design,
• documentation of the results and
• the plausibility of the design and the results for risk assessment purposes.
67
Fig. 7 Approach used to evaluate studies of effects in vivo and in vitro

for use in assessing human health risks due to exposure to nano-
and microplastic particles
Source: from reference 16 (https://creativecommons.org/licenses/by/4.0/)

All the criteria have equal weight. Thus, studies with non-zero scores against all criteria ideally represent those
that should be prioritized for risk assessment.
The approach is based on previous methods for evaluating study designs and
reporting details, such as the principles of the Klimisch score (417), guidance and
criteria used in a modified version of the ToxRTool (374) and those proposed by de
Ruijter et al. (418) for assessing the quality of ecotoxicological studies. The objective
of the NMP toxicity study assessment tool is to provide a standard procedure for
evaluating and scoring the quality of toxicity studies of relevance to human health in
a transparent approach (16). The results can thus be used to screen and prioritize a
study for the purposes of risk assessment and can also be used to provide guidance
for strengthening the quality of future studies, which is perceived to be a principal
factor for assessing human health risks.
5.1 Literature review and experimental study

evaluation
A total of 109 studies with data from in-vivo or in-vitro test
systems, representing a variety of exposure pathways and
toxicological end-points, were identified and evaluated (315,
353, 355, 362, 405, 419–522). Nearly all provided results
for a monodisperse group of plastic particles: polystyrene
68
5. Toxicological effects
spheres were tested in 67% of the studies in mammals

in vivo and 80% of bioassays in vitro, in 57% of which
NMP < 1 μm were used. The second most common plastic
particle tested was polyethylene, with results in 14% of
in-vivo and 12.5% of in-vitro studies. Limited numbers of
studies provided results for polyurethane, polypropylene, PVC,
PET, acrylic ester copolymer and nylon. While the common
use of monodisperse NMP in the studies may strengthen
understanding of toxicological mechanisms of action for
specific polymeric particles of well-defined shapes and sizes,
environmental monitoring implies that human exposure
is dominated by exposure to a heterogeneous mixture of
particles of varying shape, size and polymer composition.
It is difficult therefore to extrapolate the results for elevated
concentrations of monodisperse particles to environmentally
relevant exposure scenarios (16, 523–525).
5.1.1 Experimental studies in vivo

The results of scoring with the quantitative quality assessment scoring tool developed
by Gouin et al. (16) for all studies in mammals in vivo are summarized in Figs 8 and
9. As not all the studies were designed for quantitative assessment of human health
risk, they should be scored against each criterion for comparison with other studies
rather than simply evaluated according to their rank on the total score. This approach
is consistent with evaluations proposed by de Ruitjer et al. (418) and Gouin et al. (16),
who argued that the scores presented should not be considered value judgements
but a guide for screening and prioritizing studies for further interpretation, as well as
providing guidance to improve the design and execution of future studies.
Several experimental studies have been conducted of the effects of exposure to
NMP by inhalation (Fig. 9), most of them in rats. Five polymers have been tested, with
variable results. Intratracheal instillation of PVC measuring 0.2 to < 5 µm1 at doses
of 10–125 mg/kg bw induced various biochemical and histopathological changes
in rats (355, 488). A dose of 50 mg/kg bw PVC had acute effects on biochemical
and cytotoxic parameters and lung weight, comparable to those of crystalline
silica at 10 mg/kg bw, 2 days after exposure, with recovery to normal by day 28
(355). The effects were milder than those of quartz particles (483), and the authors
concluded that PVC powders are not cytotoxic, fibrogenic or pathogenic in rats (355).
Polyurethane foam particles (< 10 μm) induced progressive inflammation in the
airways of rats after intratracheal instillation (5 mg per animal), leading to fibrosis of
the lower respiratory tract 12 months after exposure, formation of a few scars and
papillary adenomas in four rats 18 months after exposure (353). The authors noted
that the response was typical of exposure to PM. Acrylic ester polymeric particles
(10–1500 nm; +/- 1.2 μm MMAD) were not toxic in rats after inhalation at 3 and
10 mg/m3 for 6 h/day for 5 days, with a no-observable-adverse-effect level of
1
Note that this is the geometric diameter. In an aerosol, the MMAD of these particles might be different,
which would influence the dose and location of deposition in the respiratory tract.
69
Fig. 8 QA/QC evaluation scores for 76 studies in mammals in vivo
Average scores per criterion for three elements: particle characterization, experimental study design and
applicability for risk assessment. Individual criteria are summarized in Fig. 7 and in reference 16.
Fig. 9 QA/QC evaluation scores for 76 studies in mammals in vivo

exposed by ingestion or inhalation
Average scores per criterion for three elements: particle characterization, experimental study design and
applicability for risk assessment. See references 15 and 16 for further information.
70
10 mg/m3 (475). Ethylene oxide–propylene oxide polymer particles (1.1 μm MMAD)

were highly toxic when administered to rats by inhalation, the toxicity increasing with
chain length and molecular weight. The median 4-h lethal concentration of the most
toxic compound, U-1500, was 106 (range, 4–245) mg/m3. This compound induced
pulmonary haemorrhage after repeated exposure to aerosols (6 h/day, 5 days/
week) for 2 weeks at concentrations as low as 5 mg/m3 (484), although there were
significantly fewer lesions after a 2-week recovery period. U-1500 ethylene oxide–
propylene oxide also induced biologically significant focal (0.3 mg/m3) or multi-focal
(5.2 mg/m3) fibrosis in rats after inhalation for 6 h/day, 5 days/week for 13 weeks,
with no change during a 5-week recovery period (482). Long-term exposure to U-1500
particles thus caused irreversible pulmonary fibrosis at concentrations as low as
0.3 mg/m3. Other ethylene oxide–propylene oxide polymer particles were not toxic,
causing only slight alveolitis after inhalation for 6 h/day, 5 days/week for 2 weeks at
100 mg/m3 (480).
The occurrence of “frustrated phagocytosis”2 in the distal lung has not been reported
in occupational epidemiology studies on NMP (section 3), and no association has
been found with mesothelioma, as the diameter and aspect ratios of synthetic fibre
dust are different from those of certain asbestos fibres and some carbon nanotubes.
Flock dust was found to contain fibres 10–15 µm in diameter and ~ 1000 µm long,
consisting of respirable particles and elongated shreds of nylon (aerodynamic
diameter, 4–8 µm) (314). While a proportion of the shreds were characterized as
fibres, with an aspect ratio > 3:1, they were morphologically distinct from high-aspect
ratio asbestos and carbon fibres, with heterogeneous width and surface. The main
properties common to asbestos and carbon fibres and nylon flock dust are durability
and biopersistence. It might therefore be hypothesized that the structure–toxicity
relation for synthetic fibres shifts above a certain diameter to physicochemical
pathogenicity influenced by (positive) surface charge. Thus, it is essential to
determine the surface charge of environmental synthetic fibres and how it changes
with fibre “age” and weathering, both indoors and outdoors.
The few studies of inhalation exposure suggest that the toxicity of some polymers
– but not of others of similar size – is due to properties such as relative molecular
mass, whereas other studies indicate that size proportionately influences responses.
After intratracheal instillation of polystyrene beads (0.125 or 1 mg), the smallest
particles (64 nm) caused more lung inflammation (487), indicating that surface area is
also important. Fig. 9 indicates that the studies with the highest scores and which met
the minimal number of QA/QC criteria are those of Warheit et al. (362) and Ma-Hock
et al. (475). Most of the studies of inhalation effects were assigned a score of “0” (i.e.,
of “unacceptable quality” for extrapolating the results for use in risk assessment),
as only one or two doses were tested and a threshold effect concentration was not
used. Additional concerns are lack of information on the presence of impurities and
on surface area and surface charge, which can influence toxicity (15, 16). More than
half of the 19 studies evaluated were conducted before 2000, which may indicate
that they are not pertinent for assessing the toxicity of NMP to which populations
are exposed today . Standardized methods and reference materials representative of
2
Frustrated phagocytosis occurs when phagocytic cells present in the lung cannot remove NMP, e.g.,
because they are fully loaded already or the NMP are substantially larger than those that the cell can
remove.
71
environmentally relevant NMP (section 2) should be made available for use in toxicity
studies to ensure robust analyses of implications for human health and assessment
of risk. In the interim, study reports should include thorough characterization of
the test particles, which is important for understanding the relations between the
properties of particles and toxicological end-points, and measures to ensure that the
observations are not influenced by an artefact, such as a chemical contaminant or
endotoxin.
The number of studies on the toxicity of NMP after oral exposure has increased
recently, allowing comparison with the adverse health effects of nanoparticles such as
titanium dioxide, which is widely used as a food colourant. Effects such as changes in
inflammatory response have been summarized (526), and Pinget et al. (527) reported
a significant effect of titanium dioxide on immune cells, with increased macrophages,
and effects on gut microbiota that could trigger diseases such as inflammatory
bowel disease and colorectal cancer. Results for other particle types, including NMP,
have been both similar and contrasting. Grouping and read-across approaches for
comparing the physicochemical properties of nanoparticles of different composition,
shape, size and surface chemistry have been used (447, 508, 528–532). In view of
the uncertainties of both exposure to and the toxic effects of NMP, however, findings
on the toxicity of other nanoparticles cannot be extrapolated to NMP. Nevertheless,
understanding how the properties of microparticles influence adverse health
effects could be important and should be considered in studying toxicity and in risk
assessment (533).
In its report on MP in drinking-water (1), WHO noted that there were no
epidemiological studies on ingested MP and that the data, at that time, from studies in
animal models in vivo were limited and inadequate for a risk assessment of ingested
MP. Currently, data on the absorption and toxicity of plastic particles are available
for only a few polymeric particles, i.e., polystyrene, polyethylene and PET, and the
reliability of some of the studies is doubtful (1), as discussed below. The relevance
and reliability of current data for evaluating the implications of exposure to NMP for
human health should therefore be evaluated.
Since the WHO report (1), several relevant publications on toxicity in vivo after short-
term exposure of the gastrointestinal tract to NMP have been published and reviewed,
(see, for instance, 15, 16, 27, 523, 524, 534–536). For this report, the studies were
evaluated with regard to characterization of particles, study design and applicability
for risk assessment (Figs 9 and 10). The NMP used in toxicity tests should be
characterized better to improve understanding of their toxicological mechanisms of
action (537), such as by use of standardized reference materials (15, 16, 538).
Fig. 9 summarizes the studies evaluated. In most, polystyrene particles of different
sizes, mainly in the micrometre range, were tested, and the species used were limited
to mice and rats. The toxic effects observed were mainly pathological changes in
the gut and liver and disorders of energy metabolism (27); reduced mucus secretion
and intestinal inflammation were also observed (448, 454). Dysfunction of the gut
barrier was reported by Jin et al. (455), and changes in the composition of the caecal
microflora were also found (448, 454, 455). Liver inflammation, lipid accumulation
and changes in the lipid profile were reported (455, 527) as well as changes in
72
Fig. 10 QA/QC evaluation scores for 37 studies of effects in vitro
Average scores per criterion for the three categories of particle characterization, experimental study design
and applicability for risk assessment. Individual criteria are summarized in Fig. 7.
markers of lipid metabolism (422, 539). Disorders in energy metabolism (422,

539) and in bile acid metabolism (455) were described. In a study of co-exposure
to organophosphorus flame retardants and NMP, Deng et al. (436) found that MP
aggravated oxidative stress, neurotoxicity and metabolic disorder. Rafiee et al. (437)
observed no significant change or abnormality in neurobehaviour after exposure to
NMP, and Stock et al. (405) found no or no significant inflammatory response or any
histologically detectable lesion in mice fed polystyrene NMP.
In a study on MP, about which there is some controversy, Deng et al. (422) exposed
groups of 5-week-old male ICR mice to 5 or 20 μm pristine polystyrene MP at a dose
of 0.01 mg/day (1 × 105 5-μm and 2 × 103 20-μm particles), 0.1 mg/day (1 × 106 and
2 × 104 particles) or 0.5 mg/day (5 × 106 and 1 × 105 particles), respectively, by oral
gavage for 4 weeks. No adverse effect was reported on body weight; however, the
relative liver weight increased at the high dose of each size of MP, with histological
evidence of inflammation and lipid droplet accumulation in affected livers. Biological
parameters of energy metabolism (ATP and lactate dehydrogenase), lipid metabolism
(total cholesterol and triglycerides), oxidative stress (glutathione peroxidase, catalase
and superoxide dismutase) and neurotoxic responses (acetylcholinesterase) in the
liver were affected in response to all doses of MP. Metabonomic analyses also implied
adverse effects on energy metabolism, lipid metabolism, response to oxidative stress
and response to neurotoxicity.
Braeuning (540) raised a number of concerns with respect to the study of Deng et
al. (350, 422), noting that the histological evidence of liver inflammation and hepatic
lipid accumulation in treated mice does not provide unequivocal evidence of an effect
owing to the quality of the histopathological analyses. Thus, Braeuning considered
73
that the small variations in biochemical measurements (Fig. 4 of Deng et al. (422))
might be due to the biological variance expected from five animals per group. Concern
has also been raised about the particle mass balance, with the tissue burden of kidney,
liver and gut combining to exceed the administered dose (541).
Several studies reported adverse effects on mammalian reproductive health. Luo et al.
(458) studied the reproductive effects of pristine 5-μm polystyrene MP administered in
drinking-water to groups of pregnant ICR mice at a concentration of 100 or 1000 μg/L
throughout gestation and lactation, with some of the female offspring at the high
concentration mated with untreated males to produce an F2 generation. The actual
concentrations, however, were not verified by analytical quantification. Exposure to MP
was reported to have no effect on body weight. The relative liver weight increased in
F1 offspring at both concentrations but not in dams, and hepatic triglyceride and total
cholesterol levels increased in dams and decreased in F1 mice at both concentrations.
Exposure to the high concentration resulted in changes in the caecal microflora in
both dams and F1 offspring. Changes in colon mucus secretion and ion transporter
transcription profile were also observed in exposed dams. Transcriptomic and
metabonomic analyses of liver and plasma indicated that exposure to MP can cause
metabolic disorder in offspring and that some consequences are still evident in F1
(280 days) and F2 offspring.
Luo et al. (452) studied the developmental effects of pristine 0.5-μm and 5-μm
polystyrene NMP administered in drinking-water to groups of pregnant ICR mice at
a concentration of 100 or 1000 μg/L throughout gestation. The F1 offspring were
maintained until postnatal day 42, when they were terminated for examination.
Exposure to MP had no effect on the sex ratio or survival of offspring, and no
statistically significant adverse effects were reported on body weight or liver to
body weight ratio. Liver and serum cholesterol and triglyceride levels were altered in
male offspring of exposed groups, and metabonomic analyses of serum, verified by
transcriptomic analysis of liver, indicated that exposure to MP in utero could cause
disordered fatty acid metabolism after birth, particularly with larger diameter particles.
Lack of information on dose, however, makes it difficult to interpret the significance of
some of the findings.
Several limitations should be considered when using these results for risk
assessment. Inadequate characterization of chemical impurities that may be
associated with the monodisperse type of particles used in the studies reduces
the usefulness of the results for assessing the implications for human health of
exposure to the complex, heterogeneous mixture of NMP expected to occur in the
environment. Thus, standard reference materials representative of environmentally
relevant ingested NMP should be made available, which will be possible only with
better characterization of NMP in food and beverages representative of human diets,
and methods are required to verify the dose actually delivered. As discussed in section
4, uncertainty in dosimetry poses challenges to interpretation and extrapolation of in-
vivo data in experimental animals to humans.
There is continuing debate about how the size of particles influences their intestinal
absorption and systemic biodistribution (section 4), with subsequent adverse effects
at the cellular level. Information on the biokinetics of particles could be combined with
74
quantitative structure–activity models to estimate their toxicity for risk assessment

(323).
In their review, the Norwegian Scientific Committee for Food and Environment (5)
suggested that the quality of current data on MP is insufficient to reach a conclusion
about the risk to human health. The conclusion was based on a systematic search
of literature published up to February 2019 and reports from EFSA (10), FAO (83)
and the European Commission’s Science Advice for Policy (11). In an analysis of the
latest studies, the German Federal Institute for Risk Assessment (344) concluded that
plastic particles in food cannot be assumed to pose a risk to human health, although
understanding of the toxicity of NMP after ingestion is still limited and largely
influenced by particle properties; however, the adverse effects of realistic exposure
concentrations in relation to tissue and individual susceptibility require further
research. These observations are consistent with those of other bodies (2, 4–6, 15).
There is thus a general awareness that the relevance and reliability of the available
data are insufficient for a quantitative assessment of risk.
5.1.2 Experimental study design

As mentioned above, a number of studies have reported
adverse effects on mammalian reproductive health in both
males and females. The studies addressed adverse effects
of NMP throughout the reproductive cycle, including on
the number of viable sperm in the epididymis (470), sperm
deformities and disruption of the blood–testis barrier (467),
translocation of NMP to the placenta and fetal tissues
(464, 471) and apoptosis of sperm cells with dose-related
expression of cytokines (467), which can serve as biomarkers
of underlying inflammation.
In their evaluation of 12 studies of mammalian toxicity, Coffin
et al. (15) identified a number of shortcomings, including
some mentioned here, that made them inadequate for
deriving a threshold value of NMP in drinking-water for effects
on human health. As noted above, in most studies only one
polymer type, usually polystyrene spheres, was tested, and
the observed adverse effects cannot be extrapolated to those
of the heterogeneous mixture of particles to which humans
are exposed. Another concern is the limited characterization
of the particles tested, and it is unclear whether the adverse
effects observed were due to the particles themselves or to
other factors, such as a chemical or endotoxin contaminant.
Other factors that could also influence the results were not
always well documented, making it difficult to attribute the
effects to exposure to the test particles.
To address these concerns, Coffin et al. (15) strongly
recommended use of the standard guidelines that have been
75
Table 7. Experimental study design criteria related specifically to animal

husbandry
Criterion Comment
Breeder, supplier, • Use of outbred animals is recommended when the aim of the
species and study is to assess responses in a heterogeneous population of
strain individuals.
• Use of inbred animals, which are genetically identical, is
recommended to control introduction of population variance due
to heterogeneity among individuals.
• Age is particularly important in the context of studies of
reproductive effect, as it is an indicator of maturity in relation to
specific reproductive end-points such as fertility, sperm counts and
pregnancy.
Housing • Acclimatization period: Researchers must demonstrate that the
conditions animals have been acclimatized to the experimental conditions
in order to evaluate stress-related factors, which are known to
influence reproductive end-points strongly.
• Environmental conditions: Light cycles, humidity and temperature
must be reported.
• Use of any environmental enriching materials must be reported, as
they may be a source of NMP and chemicals such as phthalates
that could influence interpretation of any observed adverse effect.
Reports should include:
• diet supplier and analysis of the diet;
• analytical composition of drinking-water;
• composition of water and food containers; and
• composition of materials used for bedding, including the
source and analysis.
• Animal group size and group housing, when groups consist of
more than one individual.
• Details of the availability of food. In most studies, it was reported
that food and water were available ad libitum, but none of the
studies considered the influence of food and water intake on the
biokinetics and biodynamics of NMP.
Route of • For instance, reports of studies of oral gavage did not indicate
exposure whether stomachs were devoid of food and/or the time since the
previous feeding.
• Vehicle
• How the particles are dispersed in the vehicle
Measurements • Body weight
• Body temperature
• Food and water consumption
• Animal behaviour
• Clinical and blood chemistry
• Specifics regarding when blood samples were taken,
including collection of blood samples before exposure to
demonstrate an experimental baseline
• Historical data on mating and pregnancy specific to the species
and strain used should be made available and referenced.
developed to study reproductive effects in mammals, such

as OECD 421, 422 and 443 (542–544). These guidelines
list key points to be considered in studies for evaluating
the effects of NMP on reproductive toxicity, from fertility to
effects on the fetus, birth and weaning, which strengthen the
reliability, relevance and comparability of the data obtained.
The systematic approach of the OECD guidelines enables
76
quantitative analysis of mammalian reproductive effects,

such as those on sperm counts and the impact of a stressor
on ovarian histology.
Consistent with the observations of Coffin et al. (15), we
raise concern about the reporting of animal husbandry
with respect to data interpretation. To ensure consistency
and to strengthen the comparability of future studies, it is
recommended that the study design criteria shown in Fig. 7
and the criteria specific to animal husbandry summarized in
Table 7 always be reported.
Consistent with the observations of Coffin et al. (15), we
emphasize the importance of reporting aspects of study
design. Robust, transparent reporting of the details of
animal husbandry is strongly recommended. In their review
of studies on bisphenol A, Thigpen et al. (545) showed that
external factors, such as the materials used (bedding, caging,
water bottles and standard rat chow), can strongly influence
interpretation of the observed effects (546) by introducing
estrogenic chemicals into the experimental test system.
Given the presence of plastic additives such as bisphenol
A and phthalates in a variety of consumer products and
their potential impact on reproductive health (547, 548), the
potential role of any other chemical stressors in the test
system that may influence the results should be thoroughly
addressed.
Stress on test animals is another important consideration.
Stress is influenced by a variety of environmental factors
and also experimental design (549–554). Test animals must
therefore be adequately acclimatized before the start of
a study. In their study on the effects of NMP on sperm in
male mice, Hou et al. (445) reported that the animals were
acclimatized before starting exposure; however, the mice
were housed singly during the acclimatization period, which
is a form of stress (555). As Hou et al. (445) did not state
how the animals were housed during the experimental phase
of the study, it is not possible to fully evaluate the potential
influence of the study design on the results. Housing animals
singly or in groups can influence hormonal levels, a particular
concern in studies of female reproduction (556–558). Any
change in housing conditions can cause stress in animals,
with negative effects on endocrine pathways, thereby
introducing uncertainty in interpretation of test results.
The route of exposure should be stated clearly. Exposure by
ingestion or inhalation should adhere to recommendations,
77
and standard methods should be used (e.g., OECD 39

for inhalation studies (559)), particularly in extrapolating
observed adverse effects to humans. The route of exposure
should ideally be consistent with exposure anticipated in
humans for extrapolation purposes. In several studies of
reproductive effects, NMP were introduced in drinking-water.
Very few studies, however, provided satisfactory information
on the homogeneity of the exposure or the stability of NMP
in drinking-water, with respective average scores of 0.57
and 0.47 (Fig. 8). For example, in their study of the adverse
effects of 10-μm polystyrene spheres on testicular tissues in
Sprague-Dawley rats, Ijaz et al. (470) used saline as a control
but used a different culture medium to disperse the particles
that were then administered by oral gavage. The authors
provided no analysis of the composition of the medium, the
dispersion of particles in the culture medium or the stability
of the particles in the exposure vehicle. Furthermore, they
gave no explanation for using a different vehicle for control
and test animals. Inconsistency among studies also includes
the reporting on the water used. Use of pure, distilled or tap
water as the vehicle to disperse NMP was reported, with
no analysis of the water for possible contaminants. The
examples given here are representative of issues that arise
when attempting to interpret study results, as various indirect
factors can influence adverse effects.
None of the studies reported the timing of administration
of NMP. This is particularly important in the context of oral
gavage, as the presence or absence of food in the stomach of
animals can influence biokinetics (15). Furthermore, details of
when and how physiological measurements are taken should
be provided, including those for body weight, behaviour and
food and water consumption. When clinical biochemical
samples are taken, details of how and when samples were
taken and how they were stored, including details of the
sample containers, should be reported, as these can indicate
whether adverse effects occurred before termination of the
study.
Ideally, animals should be necropsied one by one in a
separate room to avoid stress to other animals; furthermore,
the order of necropsy should be counterbalanced among
groups, and both physiological and biochemical samples
should be processed in a randomized manner (560, 561).
Blood samples should be collected during a defined period
(e.g., 09:00–13:00) to avoid diurnal variation in hormonal
levels, as recommended in the endocrine disruptor testing
guidelines of the United States Environmental Protection
78
Agency (562, 563). Additionally, when blood samples are

collected for hormone measurements, animals should be
moved to a holding room the day before necropsy to minimize
stress-related effects due to cage transport, which has been
known for some time to affect thyroid hormone levels (564).
In some studies, such as that of Ijaz et al. (470), blood
samples were collected and stored in sterile tubes at the
end of the live phase for further analysis; however, there
was no mention of how the blood was taken, the volume of
blood or the storage conditions, which raises concern about
the interpretation and therefore the validity of the results
reported. It is common practice, for instance, to collect
blood before initiating a study to characterize a baseline for
biochemical analysis for comparison with levels observed at
the end of the experiment. This helps to address variation in
the study population and strengthens data interpretation.
Ijaz et al. (470) used diethyl ether to anaesthetize the
animals in their experiment; however, this is not permitted
in many other countries and can influence the biochemical
analysis of, e.g., hepatic enzyme activity. As a general rule,
experiments in which animals are used for the purpose of
advancing scientific understanding must be designed on the
basis of a harm–benefit analysis (561). Particularly in studies
in mammals in vivo, researchers are ethically required to
adhere to standard test methods or to provide appropriate
justification of why standard practices were not followed.
We note that the studies evaluated were designed to consider
various questions and consequently differ in duration,
exposure dose and types of NMP used, toxicological end-
points, species and sample sizes. Nevertheless, adoption of
the three segments of the OECD test guidelines is strongly
recommended for any study of reproductive toxicity:
segment I: Fertility and general reproductive
performance, applicable to studies of both male and
female rats;
segment II: Teratology or embryo-fetal toxicity, applicable
to studies in rats and rabbits; and
segment III: Perinatal and postnatal development,
applicable to studies in rats on the effects of active
pharmaceutical ingredients during the last trimester of
pregnancy and during lactation.
In the study of Ijaz et al. (470), who used a 60-day exposure,
it is unclear whether that duration corresponds to a relevant
reproductive segment. It is likely that the duration was
79
selected to correspond to the length of one of the four cycles

of the seminiferous epithelium during the development of
mature sperm. The duration of spermatogenesis from type
A spermatogonia of stage VIII tubules is estimated to be 35
days in mice and 56 days in Sprague-Dawley rats (565–567).
As Ijaz et al. (470) used Sprague-Dawley rats, the 60-day
exposure was presumably meant to correspond to the
duration of spermatogenesis. To reduce potential ambiguity,
future study reports should clarify important elements of the
study design (560).
Non-standard methods were usually used for assessing
sperm and their motility and histological examination of
the testes and related structures, and the results are often
semi-quantitative rather than quantitative. Some of the
methods used were simply inappropriate, such as use of
a haemocytometer to count sperm (440). More accurate
methods for testicular histology are necessary to evaluate
reliably whether effects on testicular function represent a
direct response to exposure to a test material.
It is further recommended that measurements such as those
obtained from immunoassays be validated against negative
and positive controls. Validation is important for interpreting
toxicological responses, as use of controls can rule out
cross-reactivity and ensure comparison of matrix samples
with the reference standard (568). Negative and positive
control chemicals should be included in assay validation,
as recommended in guidance from the European Medicine
Agency (569) and the US Food and Drug Administration (570).
Interpretation of the results of most of the studies on the
toxicity of NMP with regard to human health is therefore
limited (15). Concerns other than those raised above include
insufficient statistical power due to small sample sizes, the
methods used to section tissues, lack of understanding
of the development of reproductive tissues and lack of
histopathological evidence of the presence of particles and
pathological processes, such as inflammation. Strengthening
QA/QC of both particle characterization and experimental
study design consistent with the recommendations in
this report are therefore critical for improving overall
understanding of the toxicity of NMP and the implications of
exposure for human health.
80
Fig. 11 QA/QC evaluation scores for 37 studies of effects in vitro

designed to reflect exposure by ingestion and by inhalation
Score per study for particle characterization, experimental study design and applicability for risk assessment.
See references 15 and 16 for additional details.
5.1.3 Experimental studies in vitro

In experimental studies in vitro, biological and mechanistic pathways are studied
under controlled conditions that cannot be achieved in vivo. The assays include those
of portal-of-entry toxicity in models of the lungs and gastrointestinal tract, non-cellular
assessment of the durability of NMP, protein interactions, complement activation
and pro-oxidant activity. In-vitro studies can be used to assess the effect of exposure
to particles on various physiologically relevant end-points, including cell viability,
inflammatory response, proliferation, necrosis and apoptosis; changes in these
parameters imply adverse effects on tissue homeostasis (571). Well-designed studies
can indicate the effects of the physicochemical properties of NMP on intercellular
uptake and toxicological mechanisms (572).
In-vitro studies are usually performed in monocellular systems. This, however,
excludes intercellular communication, whereas signalling among cells is central to
tissue and organ homeostasis. Caution is therefore warranted in extrapolating the
effects of particles in vitro to in-vivo systems (571). Toxic effects observed in vivo,
for instance, are not restricted to the expression of signalling molecules but include
the migration of inflammatory cells, changes in the vascular compartment, tissue
injury and fibrotic alterations. A combination of in-vitro and in-vivo data is therefore
necessary to elucidate the toxicological mechanisms of action of particles (537, 572).
Complex organotypic human models, such as organoids, 3D tissue culture and organ-
81
on-chip appear to be promising for assessing the toxicity of NMP and would also
reduce the use of animals in testing. A variety of tools will be required for QIVIVE to
determine the implications of human exposure to NMP (572).
Figs 10 and 11 summarize the results of in-vitro studies on the toxicity of NMP and the
models used, which include the airways (516, 519), intestine (493, 501, 512), placental
epithelium (493) and skin (573). The models of airways included both bronchial (516)
and alveolar epithelium. Nanosized polystyrene beads were used in most studies,
except in one, in which bronchial epithelial cells were exposed to polycarbonate
and ABS particles generated from 3D printer emissions (574). The particles used in
studies of the airways measured 0.025–2.0 μm, which are substantially smaller than
the MP detected and quantified in environmental samples (section 2) but relevant to
those implicated in adverse effects on human health (< 0.1 μm). Ultrafine particles
trigger inflammatory mechanisms in vitro that may play a role in chronic pulmonary
inflammation (517, 574). Particle surface area and the oxidative potential of ultrafine
particles appear to be critical parameters in the inflammatory response (533, 571).
Particle interactions with lipid mediators, such as COX2 protein, play a central role in
interference with regulatory pathways after exposure to ultrafine particles.
Confounding factors such as the presence of impurities (e.g., endotoxins) or chemicals
added to plastic should be considered when assessing the effects of NMP in vitro. Xu
et al. (513, 575) concluded that much of the toxicity of PVC in their tests was due to
leaching of chemical additives by assessing the toxicity of the original particles and
those from which chemical additives had been removed and comparing them with silica
particles at a similar dose and particle size distribution (513). The PVC particles were
less toxic than silica, which was considered to be due partly to faster clearance from
the lung. These observations are consistent with those of Pigott and Ishmael (483),
who assessed PVC powders (1–250 μm) obtained from an industrial source, α-quartz
(median diameter, 33 μm) as a positive control and polymethylmethacrylate powder as a
non-cytotoxic material. Aliquots of PVC dust suspensions were added to culture vessels
to a final concentration of 0.5 mg dust/106 cells and exposed for 2 h. Comparison of
alcohol-washed and unwashed powders indicated a cytotoxic effect of a surfactant
associated with the PVC dust, as the toxic response was mitigated when it was removed
(483). Separation of the effects of the particles and of the chemical thus supported
results obtained in vivo, in which minimal tissue damage was observed.
In-vitro studies relevant to oral toxicity are based on human intestinal cell models
(mainly Caco-2 cells), as reviewed by Yong et al. (27). In most studies, cellular uptake
of NMP was observed, but they generally had insignificant toxicity, except at high
concentrations. No significant effects on cell viability were observed after exposure to
5-μm polystyrene beads (501, 512). Stock et al. (405) reported significant loss of cell
viability only at very high concentrations of the smallest particles tested (1 μm); no
cytotoxicity was observed at any concentration of the larger particles. Similar findings
were reported by Hesler et al. (493), who found a significant increase in metabolic
activity after exposure to 46-nm polystyrene beads at 100 μg/mL and significantly
decreased metabolic activity after exposure to 0.01 μg/mL of 446-nm polystyrene
beads. Wu and colleagues (501) reported low toxicity after exposure to 0.1- and 5-µm
polystyrene particles but observed depolarization of mitochondria and inhibition of the
toxicant efflux pump, ATP-binding cassette transporter, which increased the toxicity
82
of arsenic. Exposure to 0.1- and 5-μm polystyrene beads at 200 μg/mL induced
significant generation of reactive oxygen species. In another study by Wu et al. (512),
5-µm polystyrene beads had no significant effect on superoxide dismutase, catalase
or glutathione activity or on malondialdehyde levels after exposure to 12.5–50 μg/mL
for 48 h.
Polystyrene beads (0.046–5 μm) did not adversely affect membrane integrity, even
at the highest concentrations (493, 501). Genes attributed to tight junction pathways
were found to be differentially expressed after exposure to 50 μg/mL polystyrene
beads (5 μm) (512), which suggested an effect on membrane integrity. Hesler et
al. (493) did not observe translocation, although polystyrene beads were observed
in intestinal cells and more 0.446-μm polystyrene beads were internalized than the
smaller 0.046-µm beads; no explanation was provided. Wu et al. (512) reported that
inflammatory and immune pathways were affected by exposure to 5-μm polystyrene
beads, although their conclusions are not supported by the data presented. Stock
et al. (405) found that uptake of microplastics did not affect the polarization of
macrophages or the release of chemokines.
Although all the in-vitro studies give insights into the potential toxicity of NMP, the
results are inadequate for risk assessment because of the use of unrealistically
high concentrations and testing predominantly of polystyrene beads, which are not
considered to be representative of environmental exposure. Moreover, as in the
reports of in-vivo studies, the properties of the particles tested were not adequately
described. Nevertheless, the factors that appear to determine dose-dependent
relations are particle size, surface chemistry (NH2-polystyrene was generally more
potent than neutral and COOH particles) and exposure duration (518). As most of
the studies were of nano-sized polystyrene particles, the results for environmentally
relevant MP of irregular or fibrous shape and different polymer composition could be
used to identify potentially hazardous particles. The in-vitro study with the highest
quality score was that of Choi et al. (489), who observed that differences in the shape
of polyethylene particles result in significant differences in toxicity. They found that
irregularly shaped particles with a rough surface structure have effects on cells
that include pro-inflammatory cytokine release and haemolysis, whereas spherical
particles were not severely cytotoxic at the concentrations tested.
5.1.4 Summary
Concern about human exposure to airborne NMP is
increasing, and characterization of their contribution to the
concentrations of atmospheric particles is important for
assessing the implications for human health (section 2).
In the Global Burden of Disease programme (311), it was
estimated that 4.2 million people had died prematurely in
2015 due to exposure to airborne PM. The components of PM
that represent the greatest risk to human health, however, are
poorly understood, although a contribution of NMP cannot be
excluded (111).
83
Dietary exposure implies potential concern about a variety

of end-points; however, the reliability and relevance of the
available data make it difficult to reach definitive conclusions.
Furthermore, contradictory results have been obtained
for monodisperse NMP, which are not representative of
environmentally relevant human exposure. Observations
both in vivo and in vitro are generally consistent with respect
to various biochemical responses, such as those related
to reactive oxygen species and various inflammatory
biomarkers. More research is required, however, on whether
mechanistic responses are direct physical effects in which
particles overwhelm cellular tissues by a particle overload
effect or are indirect effects, such as leaching of a chemical
contaminant or an indirect immune response.
A number of studies in which rodents were exposed to
polystyrene spheres (0.2–20 µm) showed adverse effects in
the digestive, hepatic, renal, thyroid, cardiac and reproductive
systems. In all the studies, the adverse effects consistently
included oxidative stress, altered metabolic profile and lipid
metabolism and chronic inflammation (419, 422).
The findings with various biomarkers suggest an association
with particle-mediated toxicity. If these biomarkers are
relevant to adverse outcomes, they will provide additional
insight into the mechanisms of the effects initiated or
aggravated by exposure to NMP. The in-vitro studies
evaluated, for instance, provide evidence that exposure to
NMP results in a pattern of inflammation and reactive oxygen
species. While the results of a number of studies suggest
a trend to inflammation and oxidative stress, which might
be associated with a molecular event, the QA/QC of study
design must be strengthened, particularly when different
organ and cell systems and different types of NMP are tested.
Inconsistent QA/QC, discussed above and elsewhere (15, 16),
limits definitive conclusions.
5.2 NMP as vectors of chemical exposure

NMP may present a hazard in various ways: because of their physical form; as vectors
of chemicals, including monomers, additives and sorbed chemicals; and as vectors
of microorganisms in biofilms (section 6). As risk is a function of both toxicity and
exposure, reliable, relevant characterization of both components is essential. The
leaching of chemicals from NMP has been identified in studies both in vivo and in
vitro as a factor in the observed adverse effects; therefore, attribution of adverse
effects to the particles themselves is difficult to establish if the concentrations of
chemical contaminants are not measured. The particles tested in most studies
84
were virgin polystyrene spheres. These could leach monomer residues or various
chemical additives, which, at high concentrations, could trigger adverse effects such
as inflammation and oxidative stress. To assess the implications to human health of
NMP as vectors of chemicals associated with MP, as occurs during environmentally
relevant exposure, each pathway of exposure to the associated chemicals should
be assessed, including in the diet, by inhalation and due to leaching from NMP. The
toxicity of chemicals associated with plastics does not necessarily present a risk in
drinking-water or food if the exposure is sufficiently lower than a defined margin of
exposure (MOE) or if the exposure resulting from leaching is negligible or minor as
compared with that from other sources. Quantification of exposure to both NMP and
associated chemicals is thus critical for risk assessment.
Plastic commodities contain a variety of chemical additives and unbound monomers
that can leach into water, air or, in the case of plastic packaging, into food before
consumption (576–578). As shown in Table 1, the application and use of chemical
additives varies widely according to the polymer composition and the intended use of
the plastic product. Consequently, evaluation of the role of NMP as a vector for human
exposure to chemical additives will require characterization and quantification of
their concentrations in NMP and in the diet (124). In the absence of this information,
risk has been estimated mainly by applying conservative assumptions and exposure
scenarios for qualitative assessment of the relative implications for human health
(1, 10, 11, 83, 124, 579). For instance, EFSA (10) and FAO (83) adopted a conservative
approach in estimating that a meal of mussels could result in exposure to 4 MP/g
(Table 5). For MP with a diameter of 25 μm and a density of 0.92 g/cm3, EFSA (10)
estimated an intake of 7 μg on the basis of typical consumption of a 225-g portion
of mussels. In a conservative scenario in which chemical additives and other
sorbed chemical contaminants are assumed to be present in MP at the maximum
concentrations reported and that the total mass of chemical is bioavailable after
ingestion, EFSA estimated that exposure to sorbed chemicals represents a negligible
fraction of total intake, with increases of < 0.006% in polychlorinated biphenyls
(PCBs), < 0.004% in polycyclic aromatic hydrocarbons and about 2% in bisphenol A
over those seen in other exposure pathways.
Using the approaches of EFSA and FAO in relation to mussels and estimates of human
exposure to MP in drinking-water, WHO (1) also evaluated exposure to chemicals
from ingestion of water contaminated with MP. Assumptions were made that would
result in very high exposure to total MP on a mass basis: spherical MP with a diameter
of 150 μm, a density of 2.3 g/cm3 and an exposure of 10.4 µg/L. At a default water
consumption of 2 L/day, daily intake of MP was estimated to be 85 μg. In this highly
conservative scenario, exposure to MP would be to 1.4 μg/kg bw per day for an adult
with a default body weight of 60 kg. The report noted that a more realistic estimate
would be about 0.03 µg/kg bw per day. The plastic-associated chemicals evaluated
for their implications for human health were bisphenol A, cadmium, chlordane, di(2-
ethylhexylphthalate), dichlorodiphenyltrichloroethane, hexachlorobenzene, polycyclic
aromatic hydrocarbons, polybrominated diethyl ethers and PCBs. If these plastic-
associated chemicals are present at the maximum reported concentrations in MP
and the chemicals are 100% bioavailable after ingestion, the MOEs for each chemical
suggest that their levels are of little concern for human health.
85
Section 2 summarizes data on human exposure to MP in food, beverages, drinking-

water and air. In view of the significant difficulty of quantifying human exposure to
NMP, it would be inappropriate to estimate total exposure to chemicals leached from
NMP in food, beverages and air as was done by EFSA (10) and FAO (83) for seafood
and by WHO (1) for drinking-water. Unlike previous estimates, based on specific
exposure scenarios, the inherent uncertainties associated with estimates of total
human exposure would be highly speculative and inconclusive. We thus propose
tiered approaches based on more relevant, reliable data to quantify the concentrations
of both NMP and associated chemicals for assessing total exposure from all relevant
pathways.
For example, 80–90% of all chemical plasticizers are used in a single polymer,
PVC (67, 68). Further, metal stabilizers are added to PVC, including those based on
cadmium and lead (580), which may be present at toxic levels in plastic products.
Turner and Filella (580) observed that, at the regulatory concentration limits in
electrical and electronic plastic commodities of 100 mg/kg for cadmium and
1000 mg/kg for lead, the concentrations in recycled plastic and plastic debris
in the environment could result in non-compliant levels. For example, maximum
concentrations of 6760 mg/kg cadmium and 23 500 mg/kg lead have been reported in
plastic litter (581). Quantification of exposure to NMP characterized as PVC should be
a priority, as this polymer may be the largest source of chemical additives for humans
and the environment (582). While PVC is found in about 10% of consumer plastic
products, it was also detected at 2% in plastic litter in the environment (580). The
relatively small proportion of MP from PVC is also consistent with data presented in
section 2 on potential exposure via drinking-water, food and beverages.
Turner and Filella (583) estimated a threshold limit of 8 mg/day of metal-contaminated
PVC and simulated mobilization of cadmium and lead from PVC in simulated gastric
fluids of seabirds, finding maximum exposure to 20 mg/kg cadmium and 1800 mg/kg
lead (583). Mohamed Nor and Koelmans (584) simulated mobilization of a number
of PCBs in artificial gut fluid and also uptake of chemicals by MP from contaminated
food, including the effect of digestion (585). Biphasic kinetics was observed, which
allowed derivation of kinetic rate parameters for each of the two sorption reservoirs.
Bioavailability was defined as the fraction of chemical desorbed from these reservoirs
according to the human gut retention time, estimated from the calibrated model.
Bioavailability was thus shown to depend on gut retention time.
There is growing interest in evaluating the bioavailability (or at least bioaccessibility)
of plastics-associated chemicals (582, 584, 586–593). Although use of simulated
gastric fluids, such as by Turner and Filella (583), allows quantification of
bioaccessibility, additional methods would be necessary to characterize the
bioavailable fraction (583, 591). In their assessment of the weight of evidence for MP
as vectors of plastic-associated chemicals, Koelmans et al. (591) observed that, while
many studies reported bioaccessible amounts or the leached fraction of chemicals
from MP in simulated gastric fluids, most did not consider possible interactions
with other components of the digestive system in vivo. Thus, according to the
physicochemical properties of a chemical, the leached fraction might be partitioned
into other compartments in the gut; for example, indigestible fractions of the natural
diet would be eliminated by excretion. While the bioaccessible fraction would be a
86
refinement of highly conservative assumptions derived from screening, which are

based on 100% bioavailability, evaluation of bioavailability in a more realistic exposure
scenario would improve assessment. The approach would be consistent with
standards for assessing the migration of chemical additives from consumer products,
such as EN 71-3:2019 for the migration of certain elements from toys in the European
Union (594) and methods that have been proposed for assessing the bioaccessibility
of chemicals (586, 595).
A possible framework for quantifying the bioaccessible fractions of chemicals was
proposed by Mohamed Nor et al. (124) and Mohamed Nor and Koelmans (584), who
used a probabilistic kinetics exposure model for plastic-associated chemicals, with
three components. The first is a probabilistic exposure model for 1–5000 µm MP
(see section 2.4). In the second component, given a known average particle retention
time in humans, desorption or resorption of plastic-associated chemicals in the gut is
modelled according to particle size-specific adsorption and desorption rate constants
measured independently in artificial desorption experiments of intestinal fluid in vitro
(584, 585), with dynamically changing concentration gradients. Bioavailability was
thus modelled over time. The background concentrations and fluxes of the same
chemicals in common foods were included in quantification of the concentration
gradients in the gut. The third component is a traditional PBPK model that simulates
subsequent uptake and biodistribution of bioavailable plastic-associated chemicals in
the body. Simulations were performed with and without the inclusion of NMP in food
and drinking-water, with probabilistic account of the multidimensionality of NMP and
uncertainty in model parameters. Four representative chemicals were investigated:
benzo[a]pyrene, di-(2-ethylhexyl)phthalate, PCB126 and lead. The integrated, realistic
chemical modelling approach demonstrated that, at the 50th percentile of exposure,
the concentrations of chemicals leached from NMP resulted in a negligible change
in the tissue concentrations of the four chemicals (124). This conclusion is case-
specific, and it is recommended that the framework be applied to other chemicals of
concern.
The MOE can be estimated according to the extremely conservative assumption that
100% of a chemical is bioavailable on a mass/mass basis or as the total mass of
NMP equivalent to the MOE of concern. The most sensitive MOE from the estimates
for drinking-water reported by WHO (2) was for cadmium. With this approach, it can
be assumed that concern about exposure to cadmium would increase as the mass
of NMP increases if all NMP contain the maximum concentration of cadmium. For
instance, an MOE > 10 000 can be derived from the adult median mass of 0.6 μg/
person day and an assumed maximum level of cadmium in NMP of 6760 μg/g
(581). Performing the calculation in reverse, an assumption of a maximum mass of
NMP of 17 mg/person day can be derived that would result in an MOE of cadmium
of < 1, whereas the MOEs for other plastic-associated chemicals remain > 100.
Thus, given the relative toxic potency of cadmium and its known use in plastic
products, evaluation of its bioavailability after exposure to NMP should be a priority.
Ideally, such studies should be accompanied by monitoring to quantify the polymer
composition of NMP to which humans are exposed and the amounts of cadmium
associated with environmentally relevant exposure, with appropriate methods
to evaluate both bioaccessiblity and bioavailability. As noted above, PVC may be
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the greatest source of metal additives. Li et al. (582), however, using non-targeted
analysis, found that most organic chemical leachates that migrated into simulated
gastric fluids were from PVC. An additional concern was organic chemicals leaching
from NMP originating from recycled plastic. This is an important observation, as in
most conservative approaches it is assumed that all NMP contain the maximum
amounts of various plastic-associated chemicals. Li et al. (582), however, suggest that
conservative approaches should be revised to account for differences in relative mass
and the types of chemicals in polymers. If PVC, for instance, represents a relatively
small fraction of total exposure, estimates of chemical-specific MOE that approach
values of concern should be refined to obtain more accurate estimates of implications
for human health.

Many studies have been conducted in the past few decades
to improve scientific understanding of the toxicity and
implications for human health of exposure to a variety of
natural and synthetic particles. Adverse effects associated
with exposure by both inhalation and ingestion have been
investigated. A limited subset of NMP have been assessed,
including epidemiological data on the adverse effects of
occupational inhalation of synthetic fibres, such as nylon
and plastic dusts generated from PVC and polyurethane
foam. While adverse effects, including accumulation of
macrophages, frustrated phagocytosis, decreased lung
function, interstitial lung disease and lung cancer, have
been reported, the studies have substantial limitations,
such as limited cohort size and insufficient accounting
for confounding factors. The data are also contradictory,
as several studies found no significant relation between
exposure and adverse effects. Furthermore, occupational
exposure to particles is not representative of the exposure
of the general population. Caution is thus warranted in
extrapolating results for different types of particles and
exposure concentrations associated with occupational
activities to indoor and outdoor environments.
Controlled tests of the toxicity of NMP in vivo and in vitro
after inhalation or ingestion indicate that high concentrations
of some types of NMP elicit various biochemical effects,
some of which depend on the physical characteristics
of the particle (e.g., size, shape) and others on their
chemical characteristics (e.g., solubility, surface chemistry,
composition) (508, 596, 597). Only a few types of particle
have been studied, however, consisting mainly of polystyrene
particles of various sizes and surface chemistry. Caution
should be exercised in extrapolating observations on a
homogeneous test particle to the heterogeneous mixture of
88
particles that comprise NMP to which the general population

is exposed.
To better interpret the results of toxicity testing of NMP,
studies both in vivo and in vitro of exposure by inhalation and
ingestion have been evaluated with respect to various QA/QC
criteria (16). The results show that the majority of the reports
do not provide sufficient information on the test particles,
which is critical for understanding the mechanisms of
observed adverse effects and various toxicological end-points
and the implications of NMP for human health. Inadequate
characterization of test particles can also obviate comparison
and replication of studies. For instance, adverse effects
may not be due to the test particle itself but to a chemical
contaminant or endotoxin, the relative importance of which
may differ from one supplier to another (15, 16). Therefore,
a series of reference NMP should be made available that,
ideally, represent the NMP to which humans are exposed.
Advances in characterizing and quantifying human exposure
(section 2) are essential for future toxicity testing. For
instance, most of the data currently available on exposure are
limited to MP measuring > 10 μm, while systems to test the
properties of MP to which human exposure is most relevant
are required for risk assessment. While some studies have
investigated effects in vitro with cells representative of
internal organs (liver or brain), the results should be analysed
in quantitative extrapolation models specific to NMP. There
is significant uncertainty about the absorption and systemic
bioavailability of NMP, and integrated tools are necessary to
guide and prioritize research.
Uncertainty about exposure to NMP must be reduced. Thus,
data are required to characterize and quantify the properties
(size, shape, polymer composition, surface chemistry) of
NMP in air, drinking-water, food and beverages to be used
in a probabilistic exposure assessment (124). The selection
of in-vitro and in-vivo test systems must be guided by
accurate data on exposure, with a series of well-characterized
reference NMP in relevant, robust dosimetry models.
The applicability of existing QIVIVE and PBPK models to
interpretation of test data is uncertain, and these tools should
be assessed and new models developed as necessary that
are consistent with the principles of replacement, reduction
and refinement (the “3Rs”) for humane testing in animals.
89
• Data on toxicity after inhalation or dietary exposure for

characterizing the hazard of NMP are limited to studies with
Key messages
polystyrene beads. Information is required on the effects of particle
size, shape, polymer composition and other factors representative
of environmentally relevant NMP.
• The limited hazard characterization of NMP suggests that they may
have adverse effects similar to those of other well-studied solid and
insoluble particles through similar modes of action.
• The available data are insufficient to determine whether exposure
to NMP is associated with any direct or indirect characteristic
pathology, as concern about QA/QC has been poorly accounted for
in published studies.
90
6. NANO- AND MICRO-
PLASTICS AS VECTORS OF
PATHOGENS
Microorganisms can populate numerous surface types by forming biofilms, which
contain diverse bacteria, algae, protozoans and fungi. In the environment, plastic
can provide a new surface substrate for biofilm-forming microbial communities,
often referred to as “plastispheres” (598–600). The propensity of microorganisms to
populate plastic particles depends on physical, chemical and biological factors, which
have been studied mainly in marine environments. A conditioning film consisting of
organic and inorganic substances forms within seconds around submerged surfaces
by adsorption, and this material-specific alteration of surface properties strongly
influences the composition of the colonizing microbial community (601–603).
Environmental conditions, including high nutrient concentrations (nitrogen and
phosphorus), salinity, temperature, high ultraviolet radiation and oxygen content,
influence the formation of plastics and microplastics biofilms (604–608). Material
properties such as hydrophobicity and surface roughness also affect microorganism
attachment and propagation (609–611). The increasing number of plastic surfaces
available for biofilm colonization on aquatic ecosystems is a topic of increasing
concern and research. A limited number of studies suggest that plastispheres can
disperse over longer distances than other particles, potentially introducing invasive
species into vulnerable ecosystems (612–614).
6.1 Microplastic-associated biofilms in water

Biofilms can protect microorganisms from external stressors
such as ultraviolet light and toxic substances and facilitate
nutrient accumulation and horizontal gene exchange (615).
While most microorganisms in biofilms are thought to be non-
pathogenic, they may harbour pathogens that multiply only
after they have infected a host (616).
The WHO report on MP in drinking-water (1) discussed
the hazards and potential risks associated with biofilms,
which may attach to and colonize MP and find their way into
drinking-water or drinking-water sources. Although MP may
serve as vectors for harmful organisms, including enteric
viruses and protozoa, the significance of microplastic-
associated biofilms is probably negligible because the much
larger surface area of other particles in drinking-water and
drinking-water distribution systems can attract more biofilms
than MP. A limited number of studies of fresh water suggest
that MP might function as vectors for long-distance transport
of pathogens and thus increase the transfer of organisms
91
with antimicrobial resistance. Plastic-mediated transport of

pathogens should not, however, be overestimated, as there
are significantly larger sources of opportunistic and obligate
pathogens in surface waters used as sources of drinking-
water. In addition, clarification and membrane treatment
of drinking-water remove most plastic particles, and
disinfection, including in distribution systems, can inactivate
pathogens and control their growth. The possibility that
non-pathogenic microorganisms could acquire and spread
antimicrobial resistance genes is an issue of concern and
should be studied further. WHO (1), while acknowledging
substantial lack of data, concluded that there was no
evidence of a risk to human health of microplastic biofilms
in drinking-water. Research should, however, be conducted
on horizontal transfer of antimicrobial resistance genes in
plastisphere microorganisms and in other biofilms, such as
those in wastewater treatment plants.
Since publication of the WHO report in 2019, Wu et al. (617)
conducted a comparison of biofilms associated with MP and
those on rock and leaf microparticles in river water using
high-throughput sequencing. Microplastic biofilms were
found to have not only a distinctive community structure but
also distinct patterns of enrichment of antibiotic-resistance
genes, including in two opportunistic human pathogens
(Pseudomonas monteilii and P. mendocina). This finding
underlines the importance of further research on drivers of
antimicrobial resistance associated with MP (618).
MP in the marine environment also provide a novel
surface substrate for microbial communities, including
typical aquatic colonizers such as Rhodobacteraceae
and Gammaproteobacteria (619, 620) and also a few
microplastics-specific colonizers such as Hyphomonadaceae
and Erythrobacteraceae (604). It is unclear, however,
whether the diversity of microplastic-associated microbial
communities is different from that of assemblages that
colonize natural particles and are present normally in water
(611). While some studies indicate less diversity on plastic
than on non-plastic substrates (617, 621), others found no
difference in microbial composition on natural and artificial
surfaces in the ocean but rather that the assemblage is
influenced by environmental factors (622). Most of the
microorganisms reported to be associated with MP are
non-pathogenic; however, several studies have shown
that microplastics in the marine environment can harbour
opportunistic pathogens, in particular Vibrio spp., which were
found enriched on a polypropylene particle sampled in the
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6. Nano- and microplastics as vectors of pathogens
North Atlantic Gyre (598), on plastic particle samples from the

Bay of Brest, France (623), and on MP from the North Sea and
the Baltic Sea (612). Other studies did not confirm enrichment
of potential pathogens associated with plastic (624–626).
Vibrio spp. are known to form biofilms on various substrates,
including glass, wood and other natural materials. A study of
the bacterial assemblages on polyethylene, polystyrene and
wood microparticles in the Baltic Sea showed that Vibrio were
present on MP but were more abundant on wood particles.
Environmental factors and nutrient availability appeared to
be the major drivers of microbial assemblage, rather than the
substrate itself (620). A meta-analysis of studies in marine
environments concluded that the abundance of potential
human pathogens on MP and on naturally occurring particles
might be comparable (622).
Several studies found enrichment of antimicrobial resistance
genes in microplastic-associated biofilms (627–630). Gene
transfer and metabolic functions are more extensive in
biofilms than in free-living microorganisms; however, no
well-controlled comparison has been made of antimicrobial
resistance gene enrichment in biofilms on microplastic and
on natural particles. It has been suggested that not only the
composition of the microorganisms associated with MP
but also heavy metals adsorbed on MP drive enrichment of
antimicrobial resistance genes (631).
6.2 Microplastic-associated biofilms in food

Although exposure to MP in seafood, including to any associated biofilms, is expected
to be very low (see section 2.4), many pathogens efficiently establish infection with
a very small inoculum. It is not known whether exposure to microplastic-associated
pathogens can result in established infection of aquatic species and, if they do,
through what route of exposure, and it is also unknown whether humans could
subsequently be exposed to the pathogens by ingestion of contaminated seafood.
There is no experimental evidence that microplastic-associated pathogens can
establish infection in seafood. One study showed direct transfer of Escherichia coli
tagged with green fluorescent protein from MP to the gut tissue of a coral species
in a laboratory experiment, which provides preliminary proof that MP are vectors for
pathogens (632). No alteration in microbial composition was observed, however, on
MP as compared with natural chitin microparticles after passage through the gut of
the marine mussel Mytilis edulis (633). This raises the question of whether exposure
to microplastics-associated biofilms represents a greater risk of infection than
exposure to biofilms associated with naturally occurring microparticles, which are
more abundant in aquatic environments.
Although studies are lacking on potential infection due to exposure to microplastics,
food safety regulations and risk management strategies in many countries mitigate
93
the risk of exposure to pathogens in food. Cooking of seafood inactivates pathogens,

including those associated with MP.
Little is known about the adverse effects of microplastic-associated biofilms. The
available data provide no evidence of a risk to human health of such exposure. MP
constitute only a fraction of the particles in aquatic environments that provide the
surface area for biofilm formation. The possibility of enrichment of antimicrobial
resistance genes in microplastic-associated biofilms and that MP might be vectors for
pathogen transmission should, however, be studied further.
• It is unclear whether the diversity of microplastic-associated

microbial communities, including pathogens, is different from that
Key messages
of assemblages on other types of particles.

• Research should be conducted on whether exposure to
microplastic-associated pathogens can result in established
infection (and, if so, through what route of exposure) and whether
humans could subsequently be exposed to microplastic-associated
pathogens.
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7. SUMMARY AND
RESEARCH TOPICS
7.1 Summary
Environmental monitoring of air, water and biota provides convincing evidence that
NMP are distributed across the planet. The concentrations are, however, highly
variable and are influenced by human activity. In most of the studies conducted to
date, MP have been characterized and quantified in marine and freshwater systems,
and wastewater treatment effluent has been identified as an important source of NMP
in the aquatic environment (1). Characterization and quantification of NMP in air raise
awareness about the importance of the atmospheric fate and transport of NMP as
a source for both marine and freshwater systems, for human exposure by inhalation
and for contamination of food and beverages (section 2). The available studies of
concentrations of NMP in air, food and beverages were conducted in only a few
locations for only a few food categories, resulting in only crude estimates of human
exposure. The data on foods and beverages are limited to a few product types, which
are not necessarily the main foods in human diets, and limited quantitative data are
available on exposure to the inhalable fraction of particles. Although one objective of
this report was to assess risks to human health, the available data are insufficient for
a quantitative assessment of total human exposure, as estimated intake is based on
limited data with well-known analytical limitations. As observed previously (634), the
evidence is insufficient to determine risks to human health
The findings cited in this report do not, however, imply that exposure to NMP is “safe”,
as concluded by some stakeholders (635). The limits to the reliability and relevance
of the available data for quantifying exposure to and the effects of NMP on human
health and the environment and how those uncertainties should best be addressed
in attempting to determine the presence or absence of risk have been discussed
elsewhere (634, 636–638). The constructive momentum built by widespread public
awareness and an overwhelming consensus among stakeholders that plastics do
not belong in the environment should be leveraged for transformation to a more
sustainable plastics economy. In addition to measures for better management of
plastic, such as better waste treatment, and initiatives to reduce the use of plastic,
innovations should also be encouraged in materials science, particularly with regard
to the substantial releases of NMP from plastic products used throughout commerce.
As it is clear that human exposure to NMP is ubiquitous, a reduction in exposure can
only have widespread benefits for humans and the environment.
In order to assess the risk and the implications of exposure to NMP on human health,
we collected and evaluated all the available data for an assessment of the overall
weight of evidence for a risk to human health. The shortcomings that obviated a
risk assessment included inconsistencies in the data, such as in sampling and the
experimental design of studies, and the absence of clear approaches to extrapolate
the adverse effects observed in experimental test systems with monodisperse
particles to those of the complex mixture of heterogeneous NMP present in the
95
environment. The recommendations made in this report should be perceived as

guidance to decision-makers in advancing scientific understanding and reducing the
barriers to risk assessment. For instance, better understanding of the sources of,
exposure to and effects of NMP could assist prioritization of measures for mitigation.
Exposure to NMP can occur by both inhalation and in the diet. Fig. 12 summarizes the
main routes of exposure and biokinetics that are considered important for assessing
the probable effects of exposure to NMP on human health. The data on contamination
of air with MP imply that atmospheric contamination should be better characterized
and quantified. The physicochemical properties of particles, such as size, shape and
density, influence their potential deposition in the alveolar regions of the lungs, where
their biopersistence can have adverse effects (section 5). Positively charged NP,
such as NH2-polystyrene nano-sized particles, appear to be more potent than neutral
particles of similar size. The human health effects of exposure to NMP such as PVC
dust and nylon flock are well documented in occupational epidemiological studies
(section 3), and mitigation of acute and chronic exposure to elevated concentrations
of these particles is strongly recommended. The properties of NMP in air, the fraction
that contributes to PM10 and PM2.5 and their absolute concentrations should be better
characterized in order to assess the effects of inhalation of NMP on human health.
Characterization and quantification of NMP in air would improve understanding of
potential deposition onto food and beverages, which could influence estimates of
the amounts ingested. Information is required on the probability distributions of
the physicochemical properties of deposited and ingested particles, such as size,
shape, composition and surface chemistry. Section 4 summarizes the biokinetics
of distribution, translocation, clearance and elimination and indicates that particle
size determines whether particles are absorbed across gastrointestinal epithelial
tissue and where they can be distributed in the circulatory system, as the fraction
of absorbed particles increases with decreasing size. Understanding of human
exposure to particles of different sizes in the atmosphere, food and beverages could
guide research on quantifying the extent of absorption after environmentally relevant
exposure. Particles that are not absorbed are excreted directly in faeces, but limited
research has been conducted on the passage and elimination of particles through
the gut. With better data on concentrations in air, food and beverages, mass balance
models could be developed for better understanding of human exposure to NMP.
The adverse effects of inhalation of NMP (section 5) include oxidative stress,
inflammation, lipid peroxidation, DNA damage and aggravation of underlying
effects such as asthma and chronic obstructive pulmonary disease. Experimental
studies, both in vivo and in vitro, show that adverse effects are triggered at high
concentrations. Particle size, shape and surface chemistry are important properties,
but it is not known which are the most important in determining potency. For instance,
does the effect observed represent an intrinsic property of the particle or does the
concentration-dependent response indicate physical stress on the cell or organism?
If it is the latter, are the effects reversible after elimination of exposure, or are NMP
intrinsically biopersistent, resulting in long-term effects? Better understanding is
required of the factors that influence the biodynamics and biokinetics of NMP after
exposure. The current knowledge base is insufficient to differentiate adverse effects
associated with exposure to NMP from those of particles occurring naturally in the
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7. Summary and research topics
Fig. 12 Uptake and biokinetics that influence the effects on human

health of exposure to nano- and microplastic particles
diet or inhaled. Although it is known that exposure to high concentrations of PM is

associated with respiratory effects, limited quantification of NMP in air obviates a
robust risk assessment. Thus, research to identify adverse effects that are intrinsic to
NMP would provide guidance for an NMP-specific human health risk assessment. The
available data do not allow firm conclusions on the risks to human health of inhalation
or ingestion of NMP, but, as NMP are part of the PM mixture, the health impacts will
not exceed those of PM.
The role of NMP as a vector for chemicals associated with plastics and for other
contaminants and pathogens is summarized in sections 5 and 6. Estimates based
on highly conservative assumptions suggest that exposure would have to be several
orders of magnitude higher than that from drinking-water before the MOEs of
concern for plastic-associated chemicals would be exceeded. The available data are
insufficient to conclude whether leaching of plastic-associated chemicals from NMP
represents a risk for human health. Although transport of pathogens may be minimal,
exposure to pathogens and other harmful microorganisms in food and beverages
due to inadequate hygiene or improper food handling is a well-understood risk; hence,
precautions to minimize and protect humans from exposure to pathogens in food and
beverages should also protect against contaminated NMP.
Although there are several sources of uncertainty, it is recommended that risk
management strategies for mitigating exposure to NMP be considered, as reducing
exposure is key to reducing any of the potential risks considered in this report.
97
7.2 Options for reducing exposure

As briefly summarized in section 2 and in reports from various regulatory bodies,
microplastics originate from many sources, including by degradation of larger
plastic items, and are ubiquitous. As there are only limited data on the numbers and
composition of NMP in air, water, food and beverages, the most important sources of
NMP cannot be identified.
Given the importance of degradation of discarded plastic into NMP, strategies for
better management and use of plastics are critical to minimize exposure to NMP. As
noted by WHO (1), even simple, low-cost measures can reduce the input of plastics
into the environment. The Rio Declaration (639) includes a statement about the
precautionary approach, which includes cost–effectiveness: “Where there are threats
of serious or irreversible damage, lack of full scientific certainty shall not be used as a
reason for postponing cost-effective measures to prevent environmental degradation”.
As outlined in the WHO report (1), consistent with the European Union Plastics
Strategy (640), the following measures are proposed to prevent entry of plastics into
the environment.
• Improve the economics and quality of plastic recycling.
• Curb plastic waste and littering.
• Drive innovation and investment towards circular solutions and sustainable
manufacturing practices to decrease the input of waste to the environment.
• Engage in international initiatives to minimize and eliminate plastic waste.
Key messages are summarized at the end of each section of this

and research topics
report. They are brought together here.

Key messages
Introduction:
• NMP are a heterogeneous mixture of particles and fibres of
various shapes, sizes, polymer composition, surface chemistry
and associated chemicals.
• In this report, a pragmatic definition of microplastics is used, in
which synthetic polymeric particles are < 5 mm in diameter, while
NP are particles < 1 μm in diameter.
• The properties and composition of NMP change during their life-
cycle in the environment.
Human exposure:
• Human exposure to NMP is ubiquitous and occurs by all routes.
• Information on exposure from air, drinking-water, food and
beverages is limited. Data on the characteristics of NMP and their
quantification in each of these media are necessary, with better
understanding of their sources.
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7. Summary and research topics
Observations from epidemiology:

• Evidence in the literature that inhalation or oral uptake of NMP
can affect the gastrointestinal tract or other organs apart from the
lung is limited and of inadequate quality.
• Better estimates are required of exposure of the general
population to NMP and co-pollutants by inhalation and in the diet.
Dosimetry and biokinetics:

• Physiological mechanisms for the uptake, distribution and
elimination of MP minimize tissue exposure. The probability of
uptake into the body increases with decreasing particle size.
• There is insufficient information to assess biodistribution (uptake,
retention, clearance, rate of translocation), including the likelihood
that NMP will cross biological barriers after deposition on the
epithelium or after reaching the circulation.
• Dosimetry models are available for extrapolation of results on
particle inhalation obtained in experimental animals to humans,
but they have not been evaluated or validated for NMP.
• Data on the biokinetics of NMP obtained in models in vitro cannot
currently be extrapolated to the situation in vivo.
Toxicological effects:
• Data on toxicity after inhalation or dietary exposure for
characterizing the hazard of NMP are limited to studies with
polystyrene beads. Information is required on the effects of
particle size, shape, polymer composition and other factors
representative of environmentally relevant exposure to NMP.
• The limited hazard characterization of NMP suggests that they
may have adverse effects similar to those of other well-studied
solid and insoluble particles through similar modes of action.
• The available data are insufficient to determine whether exposure
to NMP is associated with any direct or indirect characteristic
pathology, as concern about QA/QC has been poorly accounted
for in published studies.
NMP as vectors for pathogens:

• It is unclear whether the diversity of microplastic-associated
microbial communities, including pathogens, is different from that
of assemblages on other types of particles.
• Research should be conducted on whether exposure to
microplastic-associated pathogens can result in established
infection (and, if so, through what route of exposure) and
whether humans could subsequently be exposed to microplastic-
associated pathogens.
99
Several themes can be identified in the key messages and are used
to define the necessary research. Generally, the characterization
and quantification of exposure to NMP and the associated human
health effects are incomplete and insufficient for an assessment of
risk, although the potential effects of NMP on human health should
continue to be monitored. As more data become available for better
understanding of mechanisms of action and subsequent effects, it may
be possible to characterize and quantify human health risk in the future.
The basic research requirements necessary to advance scientific
understanding are listed below.
• Standard methods: Sampling and analysis of NMP in air, water,
food and beverages require robust, quality-assured methods and
suitable reference standards representative of environmentally
relevant NMP.
• Particle characterization: Quality-assured environmental
monitoring studies should be conducted to characterize the
distributions of size, shape and composition of NMP in the
environment for studies of the effects of exposure on human
health and to prepare reference standards for environmentally
relevant testing of toxicity.
• Sources of NMP: Although NMP are ubiquitous in the
environment, their sources cannot currently be accurately defined.
They include tyre and road wear particles, textiles, degradation
and fragmentation of plastic, but it is not known which source
predominates. The contributions of different factors would guide
strategies for mitigating exposure.
• Uptake and fate of both inhaled and ingested NMP: Information
on the absorption and systemic uptake of NMP is available from
only a few studies with a limited number of plastic polymers.
More information is required on the absorption, distribution and
elimination of NMP. More research should be conducted on the
influence of the food matrix on the bioavailability of ingested
particles and the efficiency of their absorption and elimination.
• Toxicology: Quality-assured experiments suitable for risk
assessment should be conducted, with adequate characterization
of exposure to the types of NMP to which humans are most
commonly exposed.
100
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136
ANNEX
Quality assurance and quality control criteria to be met to achieve
each score
The criteria are considered to have been met if they are referred to in a publication.
The table below summarizes the scores for quality, with the categories and criteria
described in the literature.
Score
Activity 2 1 0
Sampling Sampling Dust: Only a subset of the Insufficient reporting
methods • Location required criteria are of sampling methods
• Date reported (e.g., date,
• Apparatus location, materials
• Mass/area collected used); however, the
data are reproducible.
Atmospheric deposition:
• Sampler description (including
collection surface area) and whether
bulk or wet deposition collected
• Location
• Date
• Height (of sampler and site, if
appropriate)
• Sampling duration (per sample and
per campaign)
• Materials used (e.g., filtered water) in
sample collection
Suspended particulate (air):

• Location
• Sampling instrument (make, model)
• Aerodynamic size fraction
• Flow rate
• Height (of sampler and site for
atmospheric air)
• Filter substrate
• Sampling duration (per sample and
campaign)
• Date and meteorological conditions
Sampling Atmospheric deposition: typically coarse Application of Inconsistent sampling
duration resolution (e.g., 1 week or 1 month) consistent sampling duration unrelated
resolution to the best to research question
Suspended particles (air): 24 – 72 h* for of the authors’ ability, or sample type, or
low volume (16.7 L/min) sampler which is appropriate insufficient reporting
Sampling duration may differ according to address the
to the nature of the sample (e.g., if highly research question
polluted, high organic content), use of
a high- or a low-volume sampler and
research question (e.g., if interested in a
specific activity)
*72 h defined as optimum by Liu et al. (1), while

24 h is typical for collecting PM10 sample (EN
12341)
137
Score
Activity 2 1 0
Sample Atmospheric deposition: Insufficient storage Insufficient reporting
processing and Sample collection in filtered water. at room temperature
storage Store sample shortly after collection in and/or or storage
the dark at 4 °C, or filter, dry and store in
a cool, dark place Unnecessary
exposure or
Suspended particles (air): contamination risk
Transfer filter to a petri dish. Store in during transportation
cool, dark place.
Mitigation of Laboratory • Cotton laboratory coat or non- Criteria met only No precautions or
contamination preparation synthetic clothes partially, e.g., only insufficient reporting
• Equipment and laboratory surfaces wiping laboratory
wiped and rinsed surfaces and
• Plastic avoided in the protocol when equipment, not
appropriate wearing a cotton
• All apparatus used is rigorously laboratory coat
cleaned with ultrapure water and/or
filtered solvents.
• All reagents and solvents used are
filtered.
Clean air • Clean room or laminar flow cabinet Mitigation of airborne No regard for airborne
conditions • A clean room should be classified contamination by contamination, use
in accordance with ISO 14644 keeping samples only of a normal fume
and/or with an indication of the closed as much as hood or insufficient
maximum permitted airborne particle possible if negative reporting
concentration. samples were run in
parallel and examined
for contamination
Negative • Field controls collected either in Insufficient type of No negative controls
control (blanks) parallel to samples (paired) or a control, e.g., fewer or insufficient
throughout the sampling period (at than three replicates, reporting
least in triplicate), but without with no reporting of negative
exposure to air or /deposition control results with no
• Laboratory (procedural) controls indication of whether
(at least in triplicate) treated and sample data were
analysed in parallel to with actual blank corrected
samples
Sample concentrations reported should

account for controls, i.e., deduct on of
baseline microplastic count, shape and
polymer type
Sample Positive control Controls (at least in triplicate) with Insufficient type of No positive controls
purification and added microplastic particles treated at a positive control or insufficient
handling the same time as the samples and for (e.g., only part of the reporting
which the particle recovery rates are protocol is tested)
determined
Sample Dust only: If wet peroxide If no proof is provided
treatment Sieving oxidation is carried that polymers are
(if necessary; if out without cooling or not affected by the
not necessary, All sample types: digestion temperature protocol (e.g., heated
a score of 2 is Digestion of sample in a protocol exceeds 50 °C KOH > 50 °C) or/
assigned) such as wet peroxide oxidation and/ insufficient reporting
or enzymes. If another chemical was
used, the effects on different polymers
should be tested before application and
reported
All sample treatments should be

onducted at < 50 °C to prevent damage
to microplastics or changes in glass
transition temperature
138
Annex Quality assurance and quality control criteria to be met to achieve each score
Score
Activity 2 1 0
Microplastic Filter/substrate Appropriate for subsequent analysis, i.e., Quartz fibre filters Insufficient reporting
characterization composition inert, flat membrane (for direct analysis by
and application micro-spectroscopy)
for assessing or composition
human that interferes with
exposure analysis
Polymer Automated, semi-automated or rigorous Hit quality indices No polymer
identification operator-approach: < 70 % when identification
library matches; performed or
Detailed, repeatable method, including low percentage insufficient reporting
whether microparticles are analysed of suspected
directly in the sample or transferred microparticles/
to new substrate, spread of particles sample area analysed;
analysed for all samples or per filter ≥ no indication of
25% of the surface area analysed. High whether microplastics
percentage of suspected microparticles are evenly distributed
analysed, i.e., all particles for which the among samples; no
numbers of pre-sorted particles are < indication of whether
100 or ≥ 50% when particle numbers > microplastics were
100; high hit quality Indices accepted analysed directly in a
(> 70%); sample or transferred
manually
Details of library or/database included or
details of software or/programme Identification with
scanning electron
microscopy and
energy-dispersive
X-ray to distinguish
polymer from non-
polymer materials
Particle Detailed reporting, including maximum No mention of Insufficient reporting
characterization and/minimum particle size and particle minimum size or/
for human size limit of detection limits of detection
exposure
Length and diameter of fibres reported Sizes based
on suspected
Classified as fibres if aspect ratio > 3:1 (not confirmed)
microplastic)
Reference
1. Liu K, Wang X, Fang T, Xu P, Zhu L, Li D. Source and potential risk assessment of suspended atmospheric
microplastics in Shanghai. Sci Total Environ. 2019;675:462–71.
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Dietary and inhalation

ISBN 978-92-4-005460-8 (electronic version)

© World Health Organization 2022

Cataloguing-in-Publication (CIP) data. CIP data are available at http://apps.who.int/iris.

Sales, rights and licensing. To purchase WHO publications, see http://apps.who.int/bookorders.

Experts who provided insights, wrote text, offered peer review,

• Annemarie van Wezel, University of Amsterdam, The

action, including generation of reactive oxidation species and

of the toxicity of environmentally relevant NMP should

Strengthening of the evidence necessary for reliable

1.1 Background and scope

food items, their use in sterile medical devices, such as

Several reports on the effects of NMP on human health

Box 1 Definitions of nano-, micro-, meso- and macroplastic particles

or as an aggregate or an agglomerate, in which ≥ 50% of the particles in the

1.3 Composition and properties of particles

Fig. 1 Attributes of NMP to be considered in assessing both exposure

Fig. 2 Classifications of manufactured and commonly encountered

Source: adapted from references 64 and 65

instance, classification is based on structure (61–63). Other

of elastomers, such as rubbers, or modified natural or

Table 1. Average densities of commonly used polymers, the applications

• Low-density polyethylene 0.89   

• High-density polyethylene 0.96   

Sources: references 1, 62, 67 and 68

1.3.4 Surface chemistry

1.3.5 Particle size

1.3.6 Particle shape

• NMP are a heterogeneous mixture of particles and fibres of

2.1 Occurrence in drinking-water

• analytical verification of polymers associated with

Box 2 Recommendations for improving sampling and analytical

• Standard methods should be developed for sampling and analysis, which

The scoring system of Koelmans et al. (2) also includes an

Table 2. Recent studies on the numbers and characteristics of

Lower size Particles/L Particles/L Predominant Quality

Fig. 3 Concentrations of MP in drinking-water according to particle size

2.2 Occurrence in air

Coarse particles (i.e., 2.5–10 μm) are usually formed by

2.2.2 Nano- and microplastic particles in air

Characterization of NMP in air can thus contribute to overall understanding of the

selected to avoid analytical signal interference or disintegration during processing,

Table 3. Studies with a total assessment score > 10 of microplastic

Additional information presented by Wright et al. (3) and in Annex 1

inhalation of indoor air is probably an important exposure pathway; however, in view

2.3 Dermal exposure

Transdermal delivery of pharmaceuticals to optimize

the negatively charged lipids in the skin and the particles.

2.4 Occurrence in food

concentrations in the tissues of fish consumed by humans. MP are therefore ingested

Table 4. Reported numbers of microplastic or microplastic-like particles

ww, per wet body weight

Table 5. Estimated daily and annual per capita ingestion of microplastic

Fig. 4 Dietary consumption from 16 food categories derived from all

of these studies is use of databases with different definitions of MP and use of

2.5 Summary and recommendations

ensure consistency in the analysis of particles within the

• Human exposure to NMP is ubiquitous and occurs by all routes.

accumulation of macrophages with some haemorrhage and increased numbers of

3.1 Summary and recommendations