Trends in Analytical Chemistry 110 (2019) 116e128

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Trends in Analytical Chemistry

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

Micro- (nano) plastics in freshwater ecosystems: Abundance,

toxicological impact and quantification methodology
Stefan-Adrian Strungaru a, 1, Roxana Jijie a, 1, Mircea Nicoara b, Gabriel Plavan b,
Caterina Faggio c, *
a
“Alexandru Ioan Cuza” University of Iasi, Department of Research, Faculty of Biology, Bd. Carol I, 20A, 700505, Iasi, Romania
b
“Alexandru Ioan Cuza” University of Iasi, Department of Biology, Faculty of Biology, Bd. Carol I, 20A, 700505, Iasi, Romania
c
Department of Chemical, Biological, Pharmaceutical and Environmental Sciences, University of Messina, Viale Ferdinando Stagno d’Alcontres, 31, 98166
Agata-Messina, Italy

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

Article history: Plastics entering the environment will persist and continue to degrade and fragment to smaller particles
Available online 9 November 2018 under the action of various environmental factors. These microplastics (MP) and nanoplastics (NP) are
likely to pose a higher environmental impact, as well as they are more prone to adsorb organic con-
Keywords: taminants and pathogens from the surrounding media, due to their higher surface area to volume ratio.
Microplastics Little known on their characteristics, fragmentation, distribution and impact on freshwater ecosystems.
Freshwater
In order to respond to these open questions concerning the plastic particles dynamics and their envi-
Spectroscopic techniques
ronmental effects, detailed sampling strategies as well as an automated, rapid, cheap and reliable
Thermo-analytical methods
Nano plastics
analytical technique suitable for routine analysis need to be developed. The review outlines the recent
advances made on examination of the potential toxicological effects of MP and NP on freshwater biota, as
well as the available analytical methods employed to identify and quantify them in environmental
samples.
© 2018 Elsevier B.V. All rights reserved.

1. Introduction
In recent years, the aquatic contaminants resulted from
anthropogenic activities, especially contamination of marine and
freshwater environments with plastic debris have drawn the sci-
entific community and the public attention [1e12]. Due to the high
production of plastics coupled with their physic-chemical proper-
ties such as buoyancy and very slow (bio)degradation rate, as well
as with the ineffective and irresponsible waste collection and
recycling, it is only a question of time when synthetic polymers
became a global ecological and environmental problem [13e15].
Therefore, the huge quantity of different plastic types deposited in
garbage landfills without any recycling plan can be easily trans-
ported in aquatic environments as plastic debris where is frag-
mented in smaller pieces under different degradation factors [16].
While, the plastic litter composed from large pieces can be easily
removed from environment and sent to recycling process, the small
* Corresponding author.
E-mail address: cfaggio@unime.it (C. Faggio).
1
These authors contributed equally.
plastic particles (<5 mm) are almost impossible to be removed from
environmental medium. Consequently, a more serious and wide-
spread ecological impact is expected for the small plastic particles
(<5 mm).
In spite of large number, approximately 30,000 different poly-
mers registered for use in the European Union, the most common
plastics, which account for about 81% of European plastic demand
in 2016, are polystyrene (PS), poly(ethylene terephthalate) (PET),
polyurethane (PUR), poly(vinyl chloride) PVC, polyethylene (PE)
and polypropylene (PP) (Fig. 1). The polymeric materials have been
designed to cover the different needs of thousands of end products,
like packaging materials (39.9%), in building and construction
(19.7%), automotive components (10%), electronic appliances
(6.2%), household appliances and sporting equipment (4.2%) and
agricultural materials (3.3%), with the remainder including furni-
ture components, medical supplies etc. [17e19].
As a result of a great variety of polymeric materials, the chemical
composition, density, size and shape of observed plastic debris in
the environments varies significantly. The plastic particles can be
classified by their shape into pellet, fragment, fiber, film and sty-
rofoam and by their size in macroplastics (>25 mm), mesoplastics

https://doi.org/10.1016/j.trac.2018.10.025
0165-9936/© 2018 Elsevier B.V. All rights reserved.
 S.-A. Strungaru et al. / Trends in Analytical Chemistry 110 (2019) 116e128
Fig. 1. Most usual polymer types with their chemical structure and their fraction of the total European demand [19].
(between 5 and 25 mm) and microplastics (<5 mm) [16,20].
Moreover, the microplastics (MP) can be subdivided either into
1e5 mm and 20 mme1 mm according to the Guidance on Moni-
toring of Marine Litter in European Seas of the EU Marine Strategy
Framework Directive (MSFD): Technical Subgroup on Marine Litter
[21] or into 500 mme5 mm and 1e500 mm [22]. From an analytical
point of view, the size differentiation according to Hidalgo-Ruz and
co-workers [22] is more suitable, particularly the first fraction
(>500 mm) for visual presorting and single spectroscopic mea-
surement. Also, a distinction can be performed between primary
and secondary MP. Primary MP comprise the manufactured plastics
of microscopic size, for example those used in cosmetics and
exfoliating scrubs or industrial pellets that served as precursor for
manufactured plastic products. Secondary MP derived from
macroscopic debris due to the influence of mechanical forces,
oxidation and photochemical processes [16,22]. The nanoplastics
(NPs) are usually defined as plastic particles which measure less
than 100 nm [23].
Despite the large and significant literature describing the source
and impacts of microplastics and nanoplastics in the ocean [24e28]
little information exists on this problem for freshwater ecosystems
[14,17,29e31]. It is considered that freshwater and terrestrial en-
vironments are the origins and transport pathways of plastics to the
oceans [31]. In order to gain a better understanding of the negative
impact of MP on aquatic biota, the physical and chemical properties
of MP must be well characterized, the MP characteristics deter-
mining their abundance, distribution and impact in the environ-
ment. Although, the literature provides a wide range of analytical
techniques for the identification and quantification of MP in envi-
ronmental samples (e.g. water, sediments and biological tissues),
these methods are difficult to apply as routine analysis. Major
limitations of these analytical methods are the time consuming
investigation procedure, their robustness and the high cost of the
instrument. Furthermore, the heterogeneous nature of the MP still
leads to strong analytical challenges.
Present paper outlines the recent advances made on the toxi-
cological impact of micro- and nanoplastics on freshwater biota, as
well as the available analytical methods employed to identify and
quantify MP in environmental samples with their advantages and
limitations. In addition, this study can contribute to improvement
of strategies and regulations regarding the plastic contamination of
the freshwater ecosystems. More than that, any reader can judge
him(her) self and take actions in order to decrease the impact of
plastic pollution and damage to life forms.
2. Actual concerns regarding the abundance of micro- (nano)
plastics in freshwater systems
Several studies have reported the presence of MP in freshwater
systems, including river beaches, surface waters and sediments of
rivers, lakes and reservoirs all over the world. Horton et al. [32]
reported for the first time the presence of MP in a UK freshwater
117
system, including both urban and rural locations. The majority of
microplastics present in sediments were locally-derived secondary
MP (e.g. broken down from larger items) and artificial fibers
introduced by sewage effluent. According to Raman analysis, the
most common polymers included polypropylene, polyester and
polyarylsulphone.
Studies carried out on rivers show high MP flow and abundance
in the Danube river [33], in rivers Rhine and Main [34] and in both
urban compartments and surface water of Greater Paris (Seine
River) [35]. Lechner et al. [33] using stationary driftnets, followed
by a visual sorting, detected approximately 937.6 ± 8543.8 (2010)
and 55.1 ± 75.4 (2012) MP items 103 m
3 in the Austrian Danube
river. Industrial raw materials, such as pellets, flakes and spherules
represented a substantial part of the plastic debris. In a worst-case
scenario, they estimated that 4.2 t plastics day
1 would discharge
Danube into the Black Sea. A study of rivers Rhine and Main (Ger-
many) showed large amount of MP with different shapes and
polymer types on the shore sediments [34]. Analysis of the MP by IR
spectroscopy showed a large amount of PE, PP and PS. Also, this
study revealed that the concentration, composition and size dis-
tribution of MP separated from river shore sediments are broadly
comparable with the results reported for sediments from marine
habitats. Furthermore, they did not find a relationship between the
MP concentration and the population density, or proximity to in-
dustrial area and sewage treatment plants. The channel current,
channel geometry, stagnant water zones and flood events may in-
fluence the microplastic accumulation and need to be addressed.
Dris et al. [35] provided for the first time the presence of MP, mostly
fibers, in total atmospheric fallout (29e280 items m
2 day
1) in
Greater Paris. Also, higher amount of fibers were found in waste-
water (260e320  103 items m
3), while in treated effluent the MP
amount significantly decreased to 14e50  103 items m
3. In
addition, by combining different sampling approaches it is possible
to collect a higher diversity of MP from small fibers (plankton net,
80 mm mesh) to fragments and beads (manta trawl, 330 mm mesh).
Further, remote areas such as Qinghai-Tibet plateau [36], Mongolia
[14] and subalpine area of Italy [37] were affected by MP pollution, as
result of a lack of plastic waste management. For example, the
abundance of MP in the surface water of Qinghai lake, the largest lake
in China, located on the northeast edge of Qinghai-Tibet Plateau, was
between 0.05  105 to 7.58  105 items km
2, in the inflowing rivers
was between 0.03  105 to 0.31  105 items km
2, in the lakeshore
sediment was between 50 and 1292 items m
2 and in the digestive
tracts of fish samples was between 2 and 15 items in specimen [36].
Small plastic debris (0.1e0.5 mm) were more abundant in the lake
surface water, larger MP (1e5 mm) dominated the river samples,
while the PE and PP were predominant in all samples where Raman
Spectroscopy was used. Microscopic plastic debris mainly were in
sheet and fiber shapes in the lake and river water samples, but were
more diverse in the lakeshore sediment. Spatially, the higher abun-
dance of MP was measured in the central part of the lake, owing to the
transportation by the lake current. The authors assumed that the main
118 S.-A. Strungaru et al. / Trends in Analytical Chemistry 110 (2019) 116e128
pollution source with MP are tourists that left plastic garbage behind
them because around the lake is not located any industrial activity and
the population density is relatively low (9.3 persons km
2). Further,
they demonstrated that MP can be ingested by organisms.
With an MP average abundance of 2.03  105 items km
2, with a
hotspot near populated areas, Lake Hovsgol, located in a mountain
area reserve, is more polluted with MP than the more developed
and densely populated Lake Huron (2779 items km
2) and Superior
(5391 items km
2) [14,38]. The majority of MP from this study were
deemed to be secondary microplastics, resulted from the frag-
mentation and degradation of plastic household debris, such as
bags, bottles, wrappers and fishing gear.
Analyzing the MP in sediments of two beaches of the subalpine
Lake Garda (Italy), Imholf et al. [37] found 108 ± 55 items m
2 and
1108 ± 983 items m
2 on the southern and northern shores,
respectively. They assumed that the higher concentration of parti-
cles on the north shore may result from the strength of the wind. It
should be noted that the number of MP are in similar range re-
ported in the studies performed on marine beach sediments.
The fragments mainly consisted of PS, PE and PP. MP recorded a
high abundance, approximately 43  103 items km
2 in the surface
waters of the Laurentian Great Lakes of the United States [38]. Erik-
sen et al. related the high level of MP pollution with the urbanization
level and population density. In addition, part of MP < 1 mm in size
were microbeads isolated from cosmetic products.
In a recent study, Ballent et al. [39] reported the occurrence of
MP in nearshore, tributary and beach sediments, along the Cana-
dian shoreline of Lake Ontario. Nearshore sediments contained on
the average the highest abundance of MP (980 items kg
1 dry
weight), followed by tributary with 610 items kg
1 and beach
sediments with 140 items kg
1. Fragments and fibers were the most
abundant MP types, while the microbeads were minor compo-
nents. Similar to previous studies, the PE was the most common
type of polymer among the Raman analyzed samples. They
assumed that MP can originate from industrial, transportation,
construction and consumer activities and their abundance can be
related to population density.
Wastewater treatment plant effluent was also identified as an
important source of MP in the freshwater environment [40]. An
incomplete removal during the treatment (>99%) from the amount
of 3.25  106 particles h
1 (>300 mm) can result in release of 1500
particles h
1 in the environment. Majority of the material identified
in the samples collected was PE, PP and thermoset plastic. It must
be noted that these data came from a highly developed country
with strong environmental politics and significant investments in
cleaning technologies and waste management.
Based on the published results, plastic particles present in
environmental matrices, such as surface and column water and/or
sediments, can originate from primary and secondary sources, been
either directly introduced through sewage discharge/storm outlets/
tributaries (e.g. PE and PP microbeads from cosmetic products and
airblast cleaning and/or artificial fibers from washing of synthetic
textiles) or derived from the breakdown of larger plastics which
may originate from numerous and varied sources (Fig. 2). In fact,
numerous studies assume that the plastic debris found in the fresh
water systems originate from land-based sources, and the rivers
and urban tributaries are the major transport pathways of MP. In
addition, the wind and currents have an important role in the
distribution of plastic debris in the freshwater environment. In
contrast to non-buoyant materials that are transported along the
tributary riverbed, the buoyant plastics are transported on surface
waters. MP in the water samples mainly were in sheet and fiber
shapes, while in sediment samples the plastic debris were more
diverse. A large number of published works have shown an increase
of MP concentration in the vicinity of cities, tourism sites, industrial
areas and sewage treatment plants, PE, PP and PS being the most
abundant MP types. Although far less data is available compared to
marine system, several studies of river sediments have shown that
the MP concentration is broadly comparable to that reported in
marine sediments studies.
3. Interactions and toxicological effects of MP and NP on
freshwater biota
In the freshwater environment both MP and NP may have direct
and indirect toxicological effects through their uptake by various
species, as well vector effects through the adsorption of organic
contaminants, such as hydrophobic persistent organic pollutants
(POPs) and of pathogens from the surrounding media (Table 1).
Further, during the degradation process monomers and additives
can be released from the polymer matrix and cause negative effects
on organisms. The link between microplastics stability, exposure
and toxicological effects highlights the complexity of this research
domain.
Analyzing the toxicological effects induced by the mere particles
is the first step before including more complex interactions, such as
the interaction between MP and other pollutants (e.g. aromatic
hydrocarbons, polybrominated diphenyl ethers and poly-
chlorinated biphenyls).
Exposure of freshwater organisms to plastic particles, particu-
larly to NP can lead to various adverse effects, including early
mortality, inflammatory responses, inhibited growth and devel-
opment, reducing energy, low feeding activity, oxidative damage,
immunity and neurotransmission dysfunction and even behavioral
abnormality [41e48]. Furthermore, the toxicological harm toward a
given species is dependent on the duration of exposure, particles
concentration, shape, chemical composition and size, with stronger
effects for smaller particles. Due to their small size, the nano-
particles can more easily pass through biological barriers and
accumulate in tissues and organs. For instance, 5 mm PS can accu-
mulate in gills, liver and gut of zebrafish, while 20 mm PS mainly
accumulate in fish gills and gut. In addition it has been demon-
strated that PS NP exposure induced a disruption of lipid meta-
bolism in zebrafish and a false indication of satiation causing less
food intake [42]. Recent studies, have demonstrated that low
density poly-ethylene (LDPE) plastic particles alone do not induce
significant effects on zebrafish after the three-week exposure
period [49,50]. According to the study performed by Rochman et al.
[43], histological abnormalities were more present in clams
exposed to PVC and PS than to PE and PET. Also, the presence of PE
microspheres reduced the predatory performance and efficiency of
Pomatoschistus microps juveniles [47].
Most evident effects have been observed for micro- (nano)
plastics with sorbed chemicals/contaminants. Ma et al. [45] re-
ported the uptake of PS beads ranging from 50 nm to 15 mm by
Daphnia magna, and a strong relationship between size of particles
and their toxicological effects and interaction with hydrophobic
pollutants from the surrounding environment. A significant toxicity
and physical damage to D. magna was observed for 50 nm PS par-
ticles, with an EC50 of 15.13 ± 3.34 mg L
1. Furthermore, the NP can
act as act as vector for phenanthrene (Phe) by absorbing and
concentrating them on the surface. The adsorption of hydrophobic
contaminants, such as Phe and 17a-ethinylestradiol (EE2) onto
unplasticized polyvinyl chloride (uPVC) has been reported by
Sleight and co-workers [51]. The direct and indirect toxic effects of
MP and NP in zebrafish (Danio rerio) larvae locomotor activity have
been investigated by Chen et al. [41]. Results showed that MP alone
exhibited no significant effects, whereas an inhibition of larval
locomotion during the last darkness period, a reduction of larvae
body length and an increase of CAT and GPx were observed for NP
 S.-A. Strungaru et al. / Trends in Analytical Chemistry 110 (2019) 116e128
Fig. 2. Some of the major ways of the contamination with micro and nanoplastics of the freshwater ecosystems.
and NP/EE2 exposure groups. It has been illustrated that co-
exposure of adult zebrafish (Danio rerio) to NP and bisphenol
(BPA) enhances the neurotoxic effects in both the central nervous
system (CNS) and dopaminergic system [44]. In another study,
Rainieri et al. [50] showed that the food supplemented with LDPE
plastic particles with sorbed chemical contaminants induced the
most obvious effects, especially in the liver. On the other hand,
different types of polymers had shown different behavior regarding
the adsorption of chemical contaminants from the surrounding
environment [43].
Rummel et al. [52] summarized in a review the influence of
microorganisms biofilm formation on plastic microparticles and its
potential impact on environment.
The majority of the studies conducted on model organisms in
laboratory conditions used PE and PS micro- (nano) spheres pre-
viously characterized at high concentrations after short-exposure,
purchased from different companies. It must be taken into ac-
count that in the freshwater ecosystems, the plastic particles have
irregular shape and are often found in the form of fragments, fibers,
foams and films. Thus, compared with microbeads, plastic frag-
ments may have sharp edge, increasing the possibility to physical
damage of gastro-intestinal wall, gills and skin. There is a lot of
information to be revealed by future studies in order to understand
in more detail the potential adverse effects of MP and NP.
4. Analytical methods used to characterize and quantify the
microplastics from environmental samples
Due to the possible harmful effects of microplastics (MP) on the
biota level, considerable efforts have been devoted by the scientist
to the development of a convenient analytical technique that
119
allows their identification and quantification in aquatic environ-
ments and in biota. In this context, the visual identification ap-
proaches using morphological criteria (e.g. shape and color),
vibrational spectroscopic techniques and thermo-analytical
methods have drawn increasing attention with regard to analyze
MP in water, sediments and biological tissues (Table 2). However,
the wide range of synthetic polymers with various chemical
structures (aliphatic, aromatic with or without functional groups)
and with different compositions as well as the presence of additives
or contaminants still give rise to strong analytical challenges.
Analysis of MP from environmental samples requires a series of
procedure, including sampling, separation, purification, identifica-
tion and quantification [56e58]. In general, selective (visual pre-
sorting), bulk and volume-reduced sampling can be distinguished
[22]. Furthermore, this can be differentiated between sampling of
the aqueous phase (e.g. surface and column water) and the sedi-
ment phase (e.g. sample from the shoreline and the river- or
lakebed) [34,59]. Basically, the larger plastic particles (>5 mm) are
removed with tweezers and their identification and quantification
is often performed by visual approaches [14,33,34]. Recent studies
have demonstrated that the visual identification alone using
morphological criteria leads to a overestimation [60,61] or under-
estimation [58] of detections. In order to analyze the smaller plastic
fraction (<5 mm), most of the MP particles (including pellets,
spheres, fragments and fibers) were extracted from sediments
samples by density separation using sodium polytungstate (SPT,
r ¼ 1.8 kg/L) [62e64] sodium chloride (NaCl, r ¼ 1.2 kg/L)
[34,65,66], sodium iodide (NaI, r ¼ 1.8 kg/L) [67], zinc chloride
(ZnCl2, r ¼ 1.6 kg/L) [68,69] and calcium chloride (CaCl2, r ¼ 1.3 kg/
L) [70,71] as separation medium or the sample were chemically or
enzimatically purified [34,61,71e73] and subsequently
120 S.-A. Strungaru et al. / Trends in Analytical Chemistry 110 (2019) 116e128

Table 1
Overview of the potential toxicological effects of various micro (nano) plastic particles and particles-associated chemicals in freshwater species, dating from 2015 to the
present.

Studied organism Studied polymer type Size Studied Exposure Interactions and toxicological effects Reference
and organic concentration
contaminants

Danio rerio (zebrafish) uPVC 200e250 mm 400 mg L
1 96 h - no mortality; [51]

larvae vector function for Phe - no changes in behavior;
and EE2 - co-exposed chemicals were involved
in genotoxicity;
PS 45 mm 1 mg L
1 120 h after 3 h post - no significant effects [41]
PS 50 nm 1 mg L
1 fertilization - higher absorption ability for EE2;
PS þ EE2 - significant suppression of locomotor
ability;
- significant reduction of larvae body
length;
- influenced mRNA expression of
nervous system genes;
- decreased the activity of AChE;
- increased the activity of CAT and GPx;
- absorption in fish body
115 ± 28 mg g
1 ww;
LDPE <17.6 mm (90%) 5, 50, 500 mg L
1 10 and 20 days - no morphological changes; [49]
- no inflammatory changes (gill, liver,
brain, kidney and intestine);
- no evidence of necrotic tissue;
- no evidence in abnormal
development;
- no changes in the transcript levels of
genes;
- it was present in intestinal lumen;
Danio rerio (zebrafish) PS 50 nm 1 mg L
1 1e3 days - absorption in fish body [44]
adults PS þ BPA - significant up-regulation of mbp
mRNA;
- significant up-regulation of a1-
tubulin;
- dopamine level was not changed;
- AChE activity decreased in the head;
LDPE 125e250 mm 4% (in food) 3 weeks - no significant effects; [50]
LDPE þ PCBs/BFRs/PFCs - no mortality;
and methylmercury - liver is the most affected organ;
PS 70 nm, 20 mg L
1, 3 weeks - inflammatory responses; [42]
5 mm and 20 mm 200 mg L
1, - increased the oxidative stress;
2000 mg L
1 - low feeding activity;
- affected the lipid metabolism;
- reducing energy;
Pimephales promelas PC 158.7 ± 41 nm e fish blood and - activated neutrophil function; - [53]
(fathead minnow) PS 41 ± 0.2 nm plasma were phagocytosis;
extracted and - degranulation;
directly exposed - neutrophil extracellular trap release;
- increased oxidative burst;
- affected the innate immune response
of fish
Pomatoschistus microps PE 420e500 mm e 24 h - reduction of the predatory [47]
(common goby) performance and efficiency;
juvenile
Chironomus tepperi (rice PE in sediment 1e4 mm, 500 MP kg
1 10 days - the size ranged 1e54 mm [48]
midge) larvae 10e27 mm, 43 - significant decreased survival;
e54 mm 100 - reduction of adult emerged number;
e126 mm
Corbicula fluminea PET 12e704 mm 4.1 mg L
1 28 days - no influence on vitellogenin [43]
(Asian clam) PET þ PCBs expression;
PE 14e704 mm 2.8 mg L
1 - histological changes in digestive
PE þ PCBs glands;
PVC 80e704 mm 4.2 mg L
1 - histological abnormalities more
PVC þ PCBs present in clams exposed to PVC or
PS 68e704 mm 3.2 mg L
1 PS;
PS þ PCBs - severe tubular dilatation in
combination of all types;
- feeding behavior was not influenced;
- no mortality;
Gammarus fossarum PA 500  20 mm 100, 540, 2680, 24 h - ingestion positively correlated with [54]
(freshwater 13,380 fibers cm
2 exposure concentration;
amphipod) - longer exposure time was responsible
for assimilation efficiency reduction
and body mass reduction;
 S.-A. Strungaru et al. / Trends in Analytical Chemistry 110 (2019) 116e128 121

Table 1 (continued )

Studied organism Studied polymer type Size Studied Exposure Interactions and toxicological effects Reference
and organic concentration
contaminants

Daphnia magna (water PE 1 mm 12.5e400 mg L
1 96 h - both dose- and time-dependent ef- [46]
fleas) 100 mm fects were observed for 1 mm PE, with
an
- EC50 ¼ 57.43 ± 3.34 mg L
1;
- 1 mm were easily ingested in guts and
were responsible for immobilization
of daphnids;
- no uptake of 100 mm particles;
PS 50 nm PS: 2.5e50 mg L
1 48 h - 14 days - a significant dose-effect was observed [45]
PS þ Phe 500 nm Phe: 0.05 for 50 nm PS
5 mm e1.2 mg L
1 (EC50 ¼ 15.13 ± 3.34 mg L
1);
10 mm - 50 nm PS act as vector for Phe by
15 mm absorbing and concentrating them
on the surface;
- physical damage to D. magna;
Chlorella pyrenoidosa PS 0.1 mm and 1 mm 10, 50 and 30 days - algal growth inhibition for 0.1 mm size [55]
(fresh water algae) 100 mg L
1 and increased with concentration;
- decreasing of photosynthetic
parameters activity and cell wall
thickness;
- cellular wall detached from plasma
membrane;
- pyrenoid was not precisely
distinguished in cell;
- distorted and unclear thylakoids.

Abbreviations: uPVC ¼ unplasticized polyvinyl chloride, Phe ¼ phenanthrene; EE2 ¼ 17a-ethinylestradiol; PS ¼ polystyrene; CAT ¼ catalase; GPx ¼ glutathione peroxidase;
GSH ¼ reduced form of glutathione; AchE ¼ acetylcholinesterase; mRNA ¼ messenger ribonucleic acids; PP ¼ polypropylene; PE ¼ polyethylene; PVC ¼ polyvinyl chloride;
PET ¼ polyethylene terephthalate; SOD ¼ superoxide dismutase; PCBs ¼ polychlorinated biphenyls; LD-PE ¼ low density polyethylene; BPA ¼ bisphenol A;
PCBs ¼ polychlorinated biphenyls; BFRs ¼ brominated flame retardants; PFCs ¼ perfluorinated compounds.

Table 2
Brief description of the main analytical methods used to identify and quantify the MPs from environmental samples with respect to their advantages and limitations.

Method Information obtained Advantages/limitations Reference

FTIR spectroscopy chemical composition þ non-destructive [62,64]

and quantification þ fast and reliable
þ aliphatic compounds and polyesters are well detectable

size (~25 mm) and thickness (<100 mm) limited

contaminations and additives can overlap polymer bands
Raman spectroscopy þ non-destructive [58,59,62,64,
þ information about the chemical composition of the polymer and contained organic and inorganic 71,72,76,77]
fillers, pigments or dyes can be achieved
þ aliphatic and aromatic compounds, C]C compounds are well detectable
compared with FTIR
þ present lower water interference
þ a higher sensitivity to non-polar functional groups
þ is not constrained by the thickness or shape of the analyzed sample by the thickness or shape of
the analyzed sample
þ can detect particles down to 1e2 mm in size

long measurement time

polymer heating and degradation

interference of fluorescence resulting from biological, organic and inorganic contamination
Pyr-GC-MS monitor MP and þ independent of MP size and shape [60,62,78]
indicate the type of þ information about the chemical composition of the polymer and contained organic additives can
polymer be achieved

destructive

dependent on MPs mass (detection limit: pg to mg) and purity (isolated MP)

inorganic additives cannot be detected

polymers with polar subunits are difficult to correctly identify
TGA-SPE/TDS-GC-MS þ independent of MP size and shape [16,79]
þ information about the chemical composition of the polymer and contained organic additives can
be achieved
compared with Pyr-GC-MS
þ independent on sample purity
þ large sample masses (up to 100 mg)

destructive
SEM surface morphology þ high-resolution images [80e82]

requires laborious preparation steps
122 S.-A. Strungaru et al. / Trends in Analytical Chemistry 110 (2019) 116e128
concentrated on filters. Cole et al. [74] have developed a rapid
method in which a proteolytic enzyme (proteinase-K) treatment
can be applied to environmental samples to digest away biological
material without destroying any microplastics debris present.
Nuelle et al. [75] have showed that using a solution of hydrogen
peroxide (H2O2) is more effective for removing biogenic material
from sediment samples than hydrochloric acid (HCl) and sodium
hydroxide (NaOH). Similarly, Cole et al. [74] demonstrated that use
of 10 mM NaOH at 60 C can damage and discolor the MPs analyzed.
The application of acid digestion in the analysis of MPs is limited
because certain strong mineral acids, such as sulfuric aid (H2SO4)
and nitric acid (HNO3) can destroy or damage polymers with a low
pH tolerance (e.g. polyamide and polystyrene). Even using a low
concentration of non-oxidizing, mineral HCl at room temperature
proved to be inefficient, yielding to large quantities of organic
material on the filter after treatment [74].
As can be seen in Table 3, the Raman and FTIR spectroscopy have
been widely applied to determine the chemical composition of
plastic debris extracted from various aquatic and terrestrial envi-
ronments. By using spectroscopic techniques, a reliable and fast
analysis of polymer debris from environmental samples is possible.
The resulting IR spectrum reveals similar information as Raman
spectrum, the sample been characterized and identified on the basis
of the chemical structure and by comparison with known reference
spectra or by knowledge of the user. With the difference that IR
absorption depends on the change of the permanent dipole moment
of a chemical bond, while the Raman spectroscopy depends on a
change in the polarisability of a chemical bond [64]. It is very inter-
esting to note that since the vibrational energy levels are unique to
each molecule, the resulting IR and Raman spectrum provide a
fingerprint of a particular molecule [83]. An advantage of these
methods is that they are non-destructive and are suitable to identify
the smaller MP fraction directly on filter. In the case of FTIR mea-
surements, the size limited is approx. 20 mm, while Raman in-
struments can detect particles down to 1e2 mm in size. Further,
compared with FTIR spectroscopy, Raman techniques present lower
water interference, a higher sensitivity to non-polar functional
groups, it is able to measure the wavenumber range below 650 cm
1
and it is not constraints by the thickness or shape of the analyzed
sample. Raman measurements can provide additional information
about contained organic or inorganic fillers, pigments or dyes, which
are not available by micro-FTIR [71]. Still the use of Raman spec-
troscopy is far from widespread, due to drawbacks such as long
measurements time, polymer heating and degradation, interference
of fluorescence resulting from micro-biological, organic and inor-
ganic contaminations and low signal to noise ratio
[58,59,62,64,72,76,77]. Cole et al. [84] showed that by using a more
advanced equipment, such as coherent anti-Stokes Raman scattering
(CARS) microscopy, an improvement of detection may be achieved.
For analysis of MP, the FTIR is carried out in attenuated total
reflection (ATR), reflectance and transmission mode. The ATR-FTIR
spectroscopy is mainly used for the assessment of the largest size
MP fraction (>500 mm) after a visual presorting by using tweezers
or a dental explorer [34,64,85,86]. For identifying the smaller MP
fraction from environmental samples, the FTIR coupled with a
microscope, called micro-FTIR has to be applied, which can be
performed in all three modes. Unfortunately, the ATR-FTIR imaging
seems to be not feasible due to the risk that MP particles adhere to
ATR crystal during the measurement. Furthermore, this approach is
very time consuming since the ATR crystal has to be focused on
every single MP particle [66]. Moreover, the reflectance micro-FTIR
spectroscopy does not give satisfying results because polymer
surfaces reflect IR light very weakly and due to the refractive error
resulting from the superposition of directed and undirected
reflection of the IR radiation by irregular shaped particle surfaces
[72,87]. Therefore, the analyses in transmission mode could facili-
tate the identification and quantification of MP within complex
samples. Transmission mode gives high quality spectra, but re-
quires IR-transparent substrates and the investigated particles need
to be sufficiently thin (<100 mm) to avoid total absorption in the
FTIR spectrum. For this purpose, Lo € der et al. [72], tested different
commercially available filter substrates and recommended an
aluminum oxide membrane filter (Anodisc 25, Whatman, GE
Healthcare) for transmission FTIR imaging. In contrast with this
filter material that is usable only from 3800 to 1250 cm
1, the novel
silicon filter substrate (10 mm holes, circular filter area of 64 mm2)
proposed by Ka €pper et al. [76] guarantees sufficient transparency
for the broad mid-infrared region of 4000-600 cm
1. Recently, by
using focal plane array detector (FPA)-based FTIR imaging, the
analysis of MP concentrated on filters without any visual presorting
became feasible [72,87e90]. Each element on such an array rep-
resents an independent IR sensor allowing a fast acquisition of
several thousand spectra within an area by one single measure-
ment. For analysis, usually the micro-FTIR data are transformed to
false-color images on the basis of integration on specific spectral
regions, such as 1480e1430 cm
1 (CeH bending, aromatic ring
stretching), 1790e1700 cm
1 (C]O stretching) or
2980e2780 cm
1 (CeH stretching), followed by an on-screen
analysis by selecting bright areas manually and comparing the
spectra to references or available databases (Fig. 3A).
Interpretation and evaluation of the complex and voluminous
FTIR imaging data were improved by Pimpke et al. [91] using an
automated software-based data analysis followed by imaging
analysis This automated approach significantly decreases the time
demand for the interpretation of data and provides information of
the particle size, particle number and polymer type of each
detected MP (Fig. 3B). In contrast with FPA-based micro-FTIR, the
Raman microscopy is not that advanced. A semi-automated Raman
microscopy method was proposed by Fre re et al. for morphological
and chemical characterization of microplastic litter [92].
In some cases, due to the presence of impurities that can overlap
the characteristic functional groups of the MP, additional structural
information needs to be provided. Moreover, chemical weathering,
aging and biochemical processes can modify their spectra features,
making identification difficult. To close this knowledge gap, in few
studies the identification of unknown MP from complex environ-
mental samples have been possible by combination of ATR-FTIR
and Raman [64] or by using the destructive thermoanalytical
methods, such as pyrolysis gas chromatography in combination
with mass spectrometry (py-GC/MS), thermogravimetric analysis
coupled to solid phase extraction (TGA-SPE) followed by thermal
desorption gas chromatography mass spectrometry (TDS-GC/MS)
and thermogravimetric analysis coupled to differential scanning
calorimetry (TGA-DSC) [16,60,75,78,79,93].
Unlike spectroscopic techniques, the py-GC/MS is a destructive
analytical technique which thermally degrades large molecules into
many individual fragmentation substances, which can be trapped by
cooling or transferred to the GC and identified by mass spectrometry.
The polymer types are identified by comparing the resulting pyro-
grams with those obtained from the pyrolysis of standard polymers.
A considerable advantage of py-GC/MS is the simultaneous detection
of polymers types of MP particles and associated organic plastic
additives (OPAs) [60,78]. But this approach is size limited by the
diameter of the thermal desorption tubes (1.5 mm) and by the ability
to manually transfer the unknown particles in the pyrolysis tube.
According to Dekiff et al. [60], the minimum particle size required for
py-GC/MS analysis was estimated to be around 100 mm. A further
disadvantage, polymers with polar subunits (e.g. polyesters and
polyethers) are often unsuitable for py-GC/MS separation and
detection. Therefore, additional structural information can be
Table 3
Overview of the different analytical methods used to assess the chemical composition, morphology and concentration of MPs in water, sediments and biological tissues, published in the last 5 years.

Analysis method MP types Size of MP Quantity Environmental matrices Location Ref.

ATR-FTIR PE, PP, PS, PU, PA, PVC, PVAL - macroplastic (>25 mm) 1029 ± 1134 MP m
2 sediments Vavvaru Island, Lhaviyani Atoll, [76]
- mesoplastic (5e25 mm) Republic of Maldives
- large MP (1e5 mm)
ATR-FTIR PE, PP, PS, 630 mme5 mm 4000 MP kg
1 shore sediments Rhine and Main rivers in Germany [34]
PET, PVC, EA, EPDM, PA and
acrylic-based polymers
ATR-FTIR PS, PP, PE, PMMA, PVC, PU 300 mme5 mm 892,777 MP km
2 aqueous phase Rhine river [73]
ATR-FTIR PE, PA, PP, PS, PET, PEST, PU <5 mm (74%) e five different fish species North and Baltic Sea [86]
reflectance and ATR micro-FTIR PE, PP, PEPP, PEST, PAN, PS, PVC, 30e500 mm (93%) from 2175 to 672 MP kg
1 sediments Lagoon of Venice [66]
PVAL, PA, alkyd resins
transmission micro-FTIR PE PP, PA, alkyd resins <5 mm from 0.002 ± 0.001 to zooplankton samples Portuguese coastal waters [98]
0.036 ± 0.027 MP m
3
FBA-based transmission and PE, PP,PE-PP copolymer, PS, PA, 10e500 mm (98%) influent: 7216 MP L
1 waste water treatment plants Denmark [88]
reflection FTIR imaging SAN, PVC, PU, PET, EVA, PVAL, effluent: 54 MP L
1

S.-A. Strungaru et al. / Trends in Analytical Chemistry 110 (2019) 116e128

acrylate
ATR-FTIR and FBA-based PE, PP, PA, PVC, PS, PU, silicone >500 mm 0e50 MP m
3 waste water treatment plants Lower Saxony, Germany [89]
transmission micro-FTIR and PU-based coatings 3 t/year of MP
imaging PE, PP, PS, PA, SAN, PEST, PVC, <500 mm 10e9000 MP m
3
PU, PET, EVA, PVAL, ABS, PLA
reflectance micro-FTIR PE 1 mme150 mm 10e1000 ppm PE-spiked sediments Lowestoft, Spurn Point and 15 offshore [87]
sites, UK
reflectance FBA-based micro- PET 1 mm e MP-spiked wastewater samples Derby, UK [90]
FTIR imaging PE, PS, nylon-6 150e250 mm
ATR-FTIR and Raman PS, PE, PAN >500 mm e sediments near Warnemünde, Germany [64]
PE, PP, PVC, PC, PS, PTFE, PET <400 mm e Gotland Basin
micro-FTIR and micro-Raman PE, PES, PA, PP <1 mm (64%) 0.1e124.7 MP L
1 in water samples waste water treatment plants Mikkeli, Finland [99]
82% MP fibers 8.2e301.4 MP g
1 dw in sludge
18% MP particles samples
Raman PET, PP, PE, PS, PVC, PSU 1e4 mm between 18.5 ± 4.2 and 66 ± 7.7 sediments Thames basin: Leach, Lambourn and [32]
MP 100 g
1 Cut rivers
Raman PP, PE, PS, PET 1e5 mm 0.05e7.58  105 MP km
2 water samples Qinghai Lake [36]
0.03e0.31  105 MP km
2 inflowing rivers
50e1292 MP m
2 lakeshore sediment
2e15 MP per samples fish samples
micro-Raman PS, PP, PE, PC, OVC, VC/VAC <5 mm 1597-12611 MP m
3 water surface Yangtze River [82]
25e300 MP kg
1 wet weight sediments
micro-Raman PP, PE, PVC and PTFE 0.5e5 mm (90%) 100e4100 MP m
3 water samples Jiaojiang, Oujiang and Minjiang [100]
Estuaries
micro-Raman PS, PE, PP, PA, PVC <5 mm north shore: 483 ± 236 MP m
2 and sediments subalpine Lake Garda, Italy [37]
1108 ± 983 MP m
2
south shore: 8.3 MP m
2 and
108 ± 55 MP m
2
micro-Raman and ICP-MS PE, PA, PS, PET, PVC, PMMA, 50e500 mm 75 ± 134 MP m
2 sediments [71]
PTFE and pigments
micro-Raman PP, EVA, PE, PS, ABS, PA, PMMA, <5 mm 13e501 MP m
3 water samples from Eropean Coast to the North [101]
PU, PET, PVC, PTFE, Atlantic Subtropical Gyre
micro-ATR FTIR PE, PP, PS, PVC, PET, PVAc, EVA, e e sediments Warnow River [62]
py-GCMS PTFE, alkyd resins
py-GCMS combined with
thermochemolysis
py-GC/MS both polymer types (e.g. PE, PP, >100 mm e sediments North Sea island of Norderney, [60]
PET, PVC, PS, PA) and OPAs Germany
py-GC/MS PP, PVC, PE, PET, PS, PU ~1 mm (91e99%) e sediments North Sea island of Norderney, [75]
Germany
(continued on next page)

123
 124 S.-A. Strungaru et al. / Trends in Analytical Chemistry 110 (2019) 116e128

obtained by combination of py-GC/MS with thermochemolysis [93].

Abbreviations: PP ¼ polypropylene; PE ¼ polyethylene; PS ¼ polystyrene; PVC ¼ polyvinyl chloride; PET ¼ polyethylene terephthalate; PVAc ¼ polyvinylacetate; EVA ¼ ethylene-vinyl acetate; PTFE ¼ polytetrafluoroethylene;
PAN ¼ polyacrylonitrile; PC ¼ polycarbonate; PU ¼ polyurethane, PA ¼ polyamide; PVAL ¼ polyvinyl alcohol; EPDM ¼ propylene diene rubber; PEST ¼ polyester; PEPP ¼ : poly(ethylene-propylene); PMMA ¼ polymethyl
methacrylate; PSU ¼ polysulphone; SAN ¼ styrene acrylonitrile; ABS ¼ acrylonitrile butadiene styrene; PLA ¼ polylactide; PES ¼ polyester; VC/VAC ¼ vinyl chloride/vinyl acetate copolymer; PTFE ¼ polytetrafluoroethylene;
[102]
[78]

[93]

[16]

[79]

[94]
Recently, Dümichen et al. proved that this combination of thermal
Ref.

extraction with thermogravimetric analysis (TGA) on solid-phase

absorbers (twisters) and subsequent analysis of these absorbers

Godesberg, Kleve-Bimmen and Lobith),

with thermal desorption gas chromatography mass spectrometry
(TGA-SPE/TDS-GC-MS) is a suitable analytic method for unknown

aquatic system: (Rhine river Bad

terrestrial system: bio gas plant

North Sea island of Norderney,

organic species in complex environmental samples [16,79]. The

advantage of this method, in contrast to other chromatographic
Ruhr and Emscher rivers

coastal waters of China

methods, such as Py-GC-MS, is the ability to analyze the polymer
Elbe and Spree rivers
North Sea, Germany

particles in complex environmental matrices without any pre-

Neureut, Germany treatment. Separation of MP from sample by flotation, filtration
and sieving and purification by using acid, alkaline and enzymatic
Germany
Location

digestion techniques are not required. Moreover, the TGA-SPE/TDS-

GC-MS can be applied for relatively large sample masses (up to
100 mg) and allows precise identification of polymer decomposition
products with very similar chemical structure. Moreover, polymers
soil, a bio gas plant, waste water

degradation products with high molecular weight can be detected. It

fish samples spiked with MP

soil, mussels and suspended

should be noted that the polymer-specific decomposition products

two wastewater effluent
Environmental matrices

need to be different from the decomposition products of the envi-

ronmental matrix. Furthermore, the solid-phase absorbers can be
treatment plant

reused and their sensitivity can be improved by varying the absor-

Mytilus edulis

bent (the most common absorbent is PDMS).

sediments

The thermogravimetric analysis coupled to differential scanning

samples
solids

calorimetry (TGA-DSC) was successfully applied to two wastewater

effluent samples for the identification of PE [94]. It has to be noted
that the thermo-analytical methods not provide information con-
cerning the number, size and shapes of polymer microfragments, due
to the destruction of the investigated particles during the analysis.
In recent years, new and innovative approaches were developed
1.5e7.6 items/individual

to analyze MP in environmental samples. For example, a rapid-

screening approach to detect and quantify MP in environmental
samples based on selective fluorescent staining using the lipophilic
0.9e4.6 items/g

fluorescent dye Nile Red (NR), followed by density-based extraction

34% and 17%

and filtration was proposed by Maes et al. [69]. Briefly, after 30 min
Quantity

of incubation with 10 mg mL
1 NP, the dye absorbs onto MP and

makes them fluorescent when irradiated with blue light. Fluores-
e

cence emission is detected using simple photography through an

orange filter and the analysis of images allows the identification
and counting of plastic particles (>100 mm). Furthermore, due the
<250 mm (17%e79%)
>1 mm (1%e34%)

ability of NR to change color as the polarity of the solvent changes

(i.e. solvatochromic nature), a categorization of plastic nature it is
below 350 mg
Size of MP

possible (Fig. 4A). Thus, it is possible to group the MP in polar

1e20 mg

polymers (nylon and PET) and hydrophobic polymers (PE, PP and

20 mg
6 mg

1 mg

PS). Future improvements are required with the aim to optimize

e

the method for identification of the individual types of plastic.

Fuller et al. [95] developed a method based on pressurized fluid
both polymer types (e.g. PE, PP,

chlorosulfonated PE) and OPAs

NIPAM ¼ poly(N-methyl acrylamide); PC12 ¼ Poly(1-dodecene).

extraction (PFE) for determining the concentration and identify

CP, PET, PE, NIPAM, PC12
PS, PA 6, chlorinated and

MPs in complex environmental samples, such as municipal waste

and soil samples. Some of the benefits of this approach include
PET, PMMA, PA6, PC

simplicity, cost, speed and uniformity in reporting concentration

results, as well as the possibility of automatization of the extraction
component, minimizing operator skill requirements.
PP, PE, PS
MP types

A new, small-scale and portable method to separate MP from

PVC

marine sediments, using the principle of density separation has

PE

PE

PE
PS

been described by Coppock et al. [96]. The Sediment-Microplastic

Isolation (SMI) unit in combination with ZnCl solution
SEM/EDS, optical microscope

(1.5 g cm
3) is able to extract MP from sediments in a single step

pyr-GC/MS combined with

with a mean efficiency of ~96%. In addition, a novel way to rapidly

thermochemolysis

TGA-SPE/TDS-GC-MS
TGA-SPE/TDS-GC-MS

monitor MP in soil by hyperspectral imaging technology and che-

and micro-FTIR
Table 3 (continued )

mometrics has been proposed recently by Shan et al. [97]. The re-
Analysis method

sults indicated that the hyperspectral imaging technology is a

potential technique to determine the MP with particle size from 0.5
py-GC/MS

TGA-DSC
and SEM

to 5 mm on soil surface directly.

Information regarding the size, shape, surface morphology and
the elemental composition of plastic debris can be achieved by
 S.-A. Strungaru et al. / Trends in Analytical Chemistry 110 (2019) 116e128 125

Fig. 3. (A) Results of transmittance FPA-based micro-FTIR chemical imaging of the spectral region 2980e2780 cm
1 of a MP sample from plankton enriched on an aluminum oxide
filter. (a) Overview of the whole sample filter. Magnified detail of the filter with a PS particle (b, red square; c. red spectrum) and a PP particle (c, blue square; c. blue spectrum). (c)
Spectra in black are reference spectra. The color band represents the intensity of the integrated band region. The edge length of a red outlined FPA detector field is 170 mm. Reprinted
with permission from Ref. [72]. (B) Novel automated analysis of FTIR imaging data from sediment sample (a) yielding polymer type (b) and particle size (c) [91].

using scanning electron microscopy plus energy-dispersive X-ray
spectroscopy (SEM/EDS). Chlorinated polymers such as PVC could
be easily identified with SEM/EDS and the high-magnification im-
ages can provide information regarding the MP surface textures.
The plastic debris analyzed using SEM were predominately eroded
fragments of larger objects with various shapes (Fig. 4B) and sur-
face textures (e.g. grooves, flakes, cracks, fractures or pits). More-
over, diatoms and bacterial cells were the most common type of
biota that exists on their surface [80e82]. However, the SEM/EDS
cannot be applied for routine analyses because it is expensive, time
consuming and requires laborious preparation steps.
Concerning the detection and occurrence of nanoplastics (NP),
the analysis methods are in early stage of development and to our
knowledge until now no NP have been detected in environmental
matrices. Under controlled conditions in the laboratory, some
analytical methods have been shown to be feasible, such as
126 S.-A. Strungaru et al. / Trends in Analytical Chemistry 110 (2019) 116e128

Fig. 4. (A) Fluorescent index vs. static contact angle values (a measure of the surface polarity) [69] (B) SEM images of MP collected from Yangtze River, classified by their shape into
(a) fiber, (b) fragment, (c) pellet, (d) film and (e) styrofoam [82].

transmission electron microscopy (TEM) [41,44,45] to measure the
particle size distribution, SEM to investigate particle surface
morphology [49,51], dynamic light scattering (DLS) for hydrody-
namic diameter of NP [41,44] and fluorescent spectrophotometer to
investigate the particle concentration in different tissues [42].
5. Conclusions and recommendations
Contamination of freshwater ecosystems with plastic debris is
an actual concern that was worldwide reported by the recent
studies. The lack of plastic debris management and recycling, as
well as the intensive usage of plastic particles in hygiene products
were identified as the main pollution sources for MP in environ-
ment that are fragmented eventually to NP. The last seems to be
more dangerous to freshwater biota than MP based on the results
provided by scientific literature. However, there is a lack of con-
sistency concerning the concentration of MP/NP used in laboratory
experiments, in relation with those measured in environmental
samples and the types of polymers tested are often limited to only
one. It is not known how the mixture between different size groups
and material types interacts with biota. In addition, it has been not
studied the synergic effect of chronic exposure at different level of
the biological communities. Thus is important to characterize the
plastic particles in order to gain a clearer understanding of their
negative impact on the aquatic biota. We herein described the
current techniques used for MP quantification and characterization
from various sample types with their advantages and limitations. It
is necessary to note that all analytical techniques have various
advantages and limitations with respect to sample handling,
equipment and their use in routine analyses is far from widespread.
Therefore, the identification and analysis of small MP from envi-
ronmental samples in a time and cost effective way as well as their
environmental effects remain open questions and challenges for
the scientific community. Further studies should be conducted to
develop an international and standardized protocol for analysis of
small sized plastic particles from environmental samples.
 S.-A. Strungaru et al. / Trends in Analytical Chemistry 110 (2019) 116e128
Acknowledgments
This work was financially supported by a grant of the Romanian
Ministry of Research and Innovation, CCCDI-UEFISCDI, project
number 26PCCDI/01.03.2018, “Integrated and sustainable processes
for environmental clean-up, wastewater reuse and waste valori-
zation” (acronym SUSTENVPRO), within PNCDI III.
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