Marine Pollution Bulletin 100 (2015) 82–91

Contents lists available at ScienceDirect

Marine Pollution Bulletin

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

A critical assessment of visual identification of marine microplastic using

Raman spectroscopy for analysis improvement
Robin Lenz a,⁎, Kristina Enders a, Colin A. Stedmon a, David M.A. Mackenzie b, Torkel Gissel Nielsen a
a
National Institute of Aquatic Resources, Technical University of Denmark, DTU, Kavalergården 6, 2920 Charlottenlund, Denmark
b
Department of Micro- and Nanotechnology, Technical University of Denmark, DTU, Ørsteds Plads, 2800 Lyngby, Denmark

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

Article history: Identification and characterisation of microplastic (MP) is a necessary step to evaluate their concentrations,
Received 21 July 2015 chemical composition and interactions with biota. MP ≥10 μm diameter filtered from below the sea surface in
Received in revised form 8 September 2015 the European and subtropical North Atlantic were simultaneously identified by visual microscopy and Raman
Accepted 15 September 2015
micro-spectroscopy. Visually identified particles below 100 μm had a significantly lower percentage confirmed
Available online 9 October 2015
by Raman than larger ones indicating that visual identification alone is inappropriate for studies on small
Keywords:
microplastics. Sixty-eight percent of visually counted MP (n = 1279) were spectroscopically confirmed being
Small microplastics plastic. The percentage varied with type, colour and size of the MP. Fibres had a higher success rate (75%) than
RAMAN particles (64%). We tested Raman micro-spectroscopy applicability for MP identification with respect to varying
Spectroscopy chemical composition (additives), degradation state and organic matter coating. Partially UV-degraded post-
Photodegradation consumer plastics provided identifiable Raman spectra for polymers most common among marine MP, i.e. poly-
ethylene and polypropylene.
© 2015 Elsevier Ltd. All rights reserved.

1. Introduction known endocrine disruptors, which suggests that sub-lethal effects

The accumulation of plastic waste in the marine environment has
been addressed by numerous studies during the last decade (Andrady,
2003; Ivar do Sul and Costa, 2014). Recently the ubiquitous spread of
microscopic plastic particles has been of high concern as a threat to
biota. The increasing attention during the past 10 years is reflected by
the publication of more than eighty studies investigating various im-
pacts of this invisible pollutant (Hidalgo-Ruz et al., 2012; Ivar do Sul
and Costa, 2014; Rocha-Santos and Duarte, 2014; Wright et al., 2013;
Cole et al., 2011).
Several laboratory studies have observed the ingestion of
micrometre-sized plastic particles by suspension feeding organisms
such as copepods (Cole et al., 2015; Lee et al., 2013; Cole et al., 2013;
Setälä et al., 2014) and mussels (Avio et al., 2015; Wegner et al.,
2012). Although, microplastics (MP) and nanoplastics may inertly
pass the digestive system along with any natural non-digestible matter,
their associated toxic effects can become a relevant hazard (Magnusson
and Wahlberg, 2014). Through fragmentation of plastic debris to small-
er sizes, the particles' surface-to-volume ratio increases, leading to an
accelerated ad- and desorption of toxic chemicals like persistent organic
pollutants (POPs) (Lee et al., 2014; Bakir et al., 2014) or heavy metals
(Graca et al., 2014). Several POPs and plastic-associated chemicals are
⁎ Corresponding author.
E-mail address: roble@aqua.dtu.dk (R. Lenz).
and induced community changes could be a major effect.
The fragmentation of marine plastic litter through physical, chemical
and biological processes is constantly adding to the pool of MPs which
become more bioavailable and widespread with decreasing size
(Browne et al., 2007; Barnes et al., 2009). Assuming that plastic debris
disintegrates in all three spatial dimensions simultaneously, the number
of fragments produced raises to the power of 3 with decreasing particle
diameter. So far, no lower size limitation of this fragmentation has been
described in the scientific literature. Biodegradation studies suggest that
if i.e. polyolefin macromolecular chains are broken down to sizes below
500 g mol−1 the polymers are readily metabolised by microbial commu-
nities (Zheng et al., 2005; Bonhomme et al., 2003).
Despite intensive research it is yet to be shown if the current and
predicted increasing future concentrations of plastic pollution in the
ocean will result in significant alterations of marine biota and ecosys-
tems. Knowledge of plastic concentrations, spatial and temporal chang-
es, size and polymer distributions and fragmentation dynamics are a
prerequisite for understanding fate and impact of MP. Sampling and
analysis of MP become increasingly more difficult and demanding the
smaller the particles. This is one reason why a majority of microplastic
surveys used a size range from 0.3 to 5 mm (Moore, 2008;
Morét-Ferguson et al., 2010; Eriksen et al., 2013; Lattin et al., 2004), a
range where visual sorting is a legitimate and quick approach. Few stud-
ies considered particles smaller than 300 μm (Desforges et al., 2014;
Norén et al., 2014; Song et al., 2014). These studies developed or com-
pared methods for MP isolation (Cole et al., 2013; Imhof et al., 2012),

http://dx.doi.org/10.1016/j.marpolbul.2015.09.026
0025-326X/© 2015 Elsevier Ltd. All rights reserved.
 R. Lenz et al. / Marine Pollution Bulletin 100 (2015) 82–91
and polymer identification using vibrational spectroscopy (Bonhomme
et al., 2003; Goldstein, 2012; Imhof et al., 2012; Cole et al., 2013; Song
et al., 2015) or electron microscopy (Norén and Naustvoll, 2011;
Vianello et al., 2013; Fries et al., 2013) in addition to visual quantifica-
tion. The common aims in these methodological explorations are
(a) to achieve higher accuracy, precision and certainty of MP identifica-
tion by avoiding false-positive and/or false-negative misidentification,
(b) to enhance analysis results with indication of polymer types and
(c) to transfer work intensive steps from manual to automated routines
(Vianello et al., 2013).
The application of spectroscopic methods allows the discrimination
of plastic and natural particles. Spectra from absorption or scattering
phenomena are analysed for the presence or absence of characteristic
frequency bands that result from excited vibrational states in the sam-
ple material. The energy difference between an excited and ground
state is characteristic for a particular molecular structure and is reflected
in the spectral signal. Fourier transform infrared (FTIR) micro-
spectroscopy has been applied in this context and can allow polymer
identification of single particles at a minimum spot size of 10 μm
(Bhargava et al., 2003). Similarly, Raman micro-spectroscopy reveals vi-
brational information of the molecular structure of samples where mod-
ern equipment achieves horizontal and vertical resolutions below 1 μm.
It is complementary to FTIR measurements i.e. Raman spectroscopy pro-
vides a better response of non-polar, symmetric bonds while FTIR al-
lows clearer identification of polar groups.
An evaluation of FTIR in comparison to visual microscopy has shown
that misidentification of small MP can be reduced by a combination of
visual search and FTIR spectral library match (Song et al., 2015). Howev-
er, poor spectrum signal qualities can result from weathered polymers
as reported by that study. Further contaminated sample surfaces hinder
automated identification by spectral library search and match functions
and increase the required manual work during spectral analysis. Both
are relevant when identifying ocean-collected MP. In recent studies
FTIR microspectroscopy has been applied for marine MP identification
more often than Raman techniques although Raman spectroscopy pro-
vides a number of advantages such as lower minimum particle size and
the usability of the complete spectral range for identification. Further,
Raman measurements are not depending on transmission of the excit-
ing light through the sample material which allows identification of
thicker or strongly absorbing particles.
Polymers collected from the marine environment can be suspected
of having been exposed to a variety of conditions that alter the original
polymer composition. Among these environmental parameters are UV-
induced photo-degradation, thermal degradation, and biodegradation
(Andrady, 2011). A recent study assessing marine MP particles by
means of FTIR reported that oxidative break-down impeded the poly-
mer identification due to spectral changes and insufficient reference
spectra for polymer degradation products (Norén et al., 2014). It is im-
portant to investigate the effects of chemically weathered plastics on vi-
brational spectral measurements before spectroscopic methods can be
implemented systematically in S-MP surveys. Raman spectroscopy has
been routinely used to study a wide range of polymer properties
(Koenig, 2001) also in the context of marine MP identification (Imhof
et al., 2012; Zettler et al., 2013).
In this study we evaluated the applicability and limitations of Raman
micro-spectroscopy relative to visual identification.
2. Methods
2.1. Sampling and preparation
The samples used in this study originate from a trans-Atlantic cruise
conducted on the Danish research vessel R/V Dana in March and April
2014 as shown in Fig. 1. Sample collection was conducted using a closed
filter unit containing a 10 μm mesh with a preceding mesh of 300 μm
(both polyamide-6). Water was pumped from an underway water
83
intake at 3 m depth at the front port side of the ship. Meshes were re-
placed in cycles of 10 ± 5 h and sealed under clean conditions and
transported back to land for later analysis.
To prevent airborne contamination sample exposure time was
minimised. The work environment and equipment was cleaned with
deionised water and detergent before sample contact and white cotton
laboratory coats, hair protection and nitrile gloves were used. During
the time of sample exposure open water-filled petri dishes were placed
next to the work area and later analysed as blank controls. The sample
material was first removed from the meshes by resuspension in
150 ml water with 150 μl sodium dodecylsulfate solution (SDS,
150 g l− 1) in closed screw-top glasses in an ultrasonic bath for 20
min. This step was repeated if necessary until the filter showed a clean
appearance. After resuspension the complete sample content was col-
lected on 10 μm vacuum filters, polycarbonate (Whatman, 25 mm di-
ameter) or polyamide (filter mesh material). Possible sample residues
were removed from the original filter by rinsing with microfiltrated
water into the vacuum filtration cylinders. In case of high material con-
tent samples were reduced using a plankton splitter.
2.2. Microscopic and spectroscopic analysis
The vacuum filters were analysed under an optical microscope
coupled to a DXR Raman spectrometer (Thermo Fisher Scientific),
with a 455 nm excitation laser and a grating of 1200 lines per mm
using 10 × and 50 × objectives (Olympus M Plan, NA = 0.25 and LM
Plan, NA = 0.50). Half of the area of the filters was searched for MP.
Prior to Raman measurements items on the filters were either accepted
or rejected as MP suspects based on a catalogue of morphological
criteria. Each accepted object was classified as one of two shape catego-
ries, fibre or particle, and subsequently photographed and measured in
length and width. Replicate Raman spectra (n ≥ 3) were acquired as
point measurements on different spots of each accepted particle in
order to eliminate the influence of local impurities such as organic mat-
ter or mineral particles.
A library of references of the major commodity consumers plastic
polymers (Table 1) was created and their Raman spectra were used as
reference for identification of MP.
To account for a broader range of chemical composition within the
polymer types, several polymer samples differing in features like flexi-
bility, colour or form, e.g. fibrous, solid, film, foamed, were included in
the library. Expected non-plastic contaminants and organic materials
present in the marine environment i.e. cellulose, methylcellulose, cellu-
lose acetate, viscose rayon, keratin and aragonite were also included
into the reference library to improve the exclusion of non-plastic (but
plastic resembling) particles. The primary reference library consisted
of a total of 52 spectra.
Raman shifted spectra were measured with a resolution of
0.96 cm−1 in a wavenumber range of 100 to 3500 cm−1 using an inte-
gration time of 2 s averaged over 10 scans. The excitation power was 3
mW at an estimated spot size of 0.5 μm. As fluorescence can occur as a
major source of poor Raman signal quality the above mentioned stan-
dard settings were changed if necessary with respect to laser power as
well as the number and duration of exposures, to enhance the signal
quality of fluorescent or weak Raman-active materials.
2.3. Data analysis
Measured Raman spectra were compared against the library by cal-
culating the Pearson correlation coefficients between each possible
combination of sample and library spectra. All spectra were
preprocessed in the following order. The wavenumber range was
cropped to 3400 to 200 cm−1. Elevated baseline for instance from fluo-
rescence signals were removed by fitting and subtracting a modelled
baseline curve which was interpolated from 25 local minima based on
a shape-preserving piecewise cubic interpolation. The spectra were
84 R. Lenz et al. / Marine Pollution Bulletin 100 (2015) 82–91

Fig. 1. Cruise map with grey sections showing the transects where water was filtered from the underway intake. Plastic concentrations are reported in an accompanying paper (Enders
et al., 2015). The stations numbered and highlighted in blue are the ones that have been used in the present study for the comparison of visual and Raman identification.

smoothed using an Savitzki-Golay non-derivative algorithm on a commercial Raman libraries (BioRad-KnowItAll Informatics System
28 cm−1 window with a second order polynomial. Each spectrum was 2014, Raman ID Expert and Thermo Fisher Scientific Inc.). Spectral
normalised to the maximum. band assignments for chemically degraded polymers were taken from
After a first run spectra from MP samples that achieved a high (Lagaron et al., 1999).
correlation (Pearson r2 ≥ 0.8) with one of the polymer references were
individually examined and, in case of confirmation, used as a
2.4. Degradation effects
secondary reference for the respective polymer type. This method
allowed to expand the coverage of the spectral library to a larger
To identify photodegraded polymers we experimentally simulated
degree of chemical variance within the target polymers.
accelerated weathering. The polymer pieces that had been used for
The final identification of MP was based on individual evaluation of
the library creation were exposed to UV light in three different treat-
each spectrum. The replicates of a sample were plotted with their best
ments. For weak degrading effects samples of LDPE, PA, PET and PVC
matching library spectra and analysed together for peak alignment
were exposed in sealed quartz cuvettes filled with (a) deionised water
and height. The pivotal criterion for acceptance was if the characteristic
and (b) filtered seawater of 32 PSU.
bands of the suspected polymer could be found in the sample spectrum.
The cuvettes were installed inside a rotating cylinder placed in front
The calculated correlation values r2 were used as an additional indicator.
of a UV light source (2 x Q-Lab UV Q-Panel 80 W). Additionally a third
All spectral processing, calculations and comparison were conducted in
series was air- exposed to the same UV light source in close proximity
Matlab (MathWorks Inc.). In unclear cases the raw spectra were
to the panels. Light intensities were measured using an NIST calibrated
revisited and unexplained bands were investigated by literature
Ocean Optics Jaz spectrophotometer and are shown in Fig. 2.
(Lambert, 1987; Edwards, 2005; Socrates, 2004) and compared against
The intensity maximum of the light source is at 343 nm. The air-
exposed samples were exposed to a 4.6 times higher intensity com-
Table 1 pared to those in water. The absorbance effect of water has been
Names and abbreviations of polymers used in the spectral identification with number of
accounted for by an attenuation coefficient of μ = 0.2. The temperature
reference items included in the library.
measured at the air-exposed and water submerged samples was ap-
Abbr. Polymer # proximately 33 °C and 22 °C, respectively.
HDPE High density polyethylene 5
LDPE High density polyethylene 4
PP Polypropylene 6 2.5. Statistical analysis
PET Polyethyleneterephthalate 5
PS Polystyrene 4 The rates of success of the visual identification, i.e. the proportion of
PVC Polyvinylchloride 3
visually identified microplastics that could be confirmed being plastic
PU Polyurethane 1
PA Polyamide 3 by Raman spectroscopic analysis, were calculated for different groups
ABS Acrylonitrile butadiene styrene 6 of MP size, colour and type. The differences between the groups were
PMMA Polymethylmethacrylate 2 tested for statistical significance by means of a two-tailed z-test for sig-
PC Polycarbonate 2 nificance of difference between two independent proportions, with
PTFE Polytetrafluoroethylene 2
p b 0.05 if not stated otherwise.
 R. Lenz et al. / Marine Pollution Bulletin 100 (2015) 82–91
Fig. 2. Spectral intensities in the UVA/B wavelength range of the UV source used to simu-
late accelerated weathering in synthetic polymers exposed in air (grey dashed) and water
(grey dotted). The black graph shows the intensity of midday sunlight (clear sky,
Denmark) in the same wavelength range.
3. Results
3.1. Comparison of visual microscopy to Raman micro-spectroscopy
A total of 452 fibres and 827 particles were visually identified as
plastic of which 75% and 64% were confirmed by Raman spectra, respec-
tively. The whole MP data set had a success rate of 68% (n = 1279). The
number of Raman-confirmed MP depended on size. Visually identified
fibres and particles of larger sizes were confirmed being plastic to a
higher degree than smaller sizes (Fig. 3).
Sixty three percent of particles b50 μm were confirmed as plastic
whereas in the group between 50 and 100 μm a slightly higher propor-
tion of 67% was confirmed. Together,they had significantly lower suc-
cess rates than the largest size class of particles N100 μm where 83%
could be successfully identified as plastic (p b 0.05). The success rate
of the whole size spectrum was not significantly different (p = 0.54)
from that of the smallest size class, as the particles b 50 μm contributed
over three quarters of total particle abundance. Fibres showed the same
trend as particles. When grouped into length categories of b1000,
1000–2000 and N2000 μm success rates were 73%, 76% and 90%,
respectively.
The success rate of the visual identification, was highest for particles
of blue colour, followed by green and red with 86%, 54% and 52%, respec-
tively as shown in Fig. 4.
The distinction of those colours was made by subjective visual obser-
vation. It could be observed that misidentification is more likely among
particles of colours that appear more frequently among natural organic
matter. Particles of transparent/white or black/grey colour which have
been visually counted for having a plastic like appearance had a low
confirmation in Raman of 41% and 42%, which were significantly
lower than for coloured particles (p b 0.001). Fibres of all colours had
a higher success rate than particles, which can be attributed to the fact
that fibres often provide more structural features for identification
(Fig. 4). In contrast to particles there was a high confirmation rate of
transparent and black fibres of 85% and 67%, respectively.
3.2. Raman signal confounding factors
We tested Raman micro-spectroscopy for its applicability in
microplastic identification studies. Commercial plastic is, apart from
its basic polymer matrix also composed of a range of additive com-
pounds. Additionally, plastic from the marine environment can have
85
Fig. 3. Microplastic particles identified by visual microscopy (whole bars) compared to
Raman spectroscopy (hatched fractions). The top graph shows the relative success rate
of visual identified particles approved by Raman spectroscopy. Number of particles com-
pared in each size class: nb50 = 637, n50–100 = 155, nN100 = 35.
an altered chemical fingerprint. Here we present the most common rea-
sons that can lead to a falsified polymer type signal and must therefore
be borne in mind.
Fig. 5 shows the spectra obtained from differently coloured ABS sam-
ples all from the same manufacturer. Blue, white and black samples
(only blue shown) were well correlating with the spectra of the type
reference (ABS ref.). The most characteristic peaks for ABS were be-
tween 2900 and 3060 cm− 1 indicative of the C\\H stretch, at
2236 cm−1 assigned to the nitrile stretching (C☰N) and the most in-
tense peak at 1001 cm− 1 which displays the vibrational response of
the aromatic ring in the styrene part of the polymer (Lambert, 1987).
Red and yellow ABS show very different peaks of noticeably higher in-
tensities. These represent a typical case of a complete overlay of the ac-
tual polymer type spectra due to a more intense compound in the
polymer matrix such as additives, in this case colour pigments. The
red sample additionally shows a broad Raman signal in the range of
2200 to 3200 cm−1 which is due to fluorescence.
A frequently encountered observation was that various morpholog-
ically discriminative black particles displayed spectra matching with the
reference spectra obtained from a set of black rubber of various sources
such as from car or bike tires or tubes (Fig. 6).
By using a spectroscopy software (Bio Rad-KnowItAll) with access to
commercial Raman libraries the two bands at 1343 cm− 1 and
1581 cm− 1 were attributed to carbon. Relative peak height had a
small variance among samples. The particle in (c) highly correlated
with PP which, owed to the morphological similarities to (a), gives an
indication of the difficulties in purely visual identification and the low
success rate obtained in the comparison study. It is not surprising to
find the presence of carbon in our sample set, as carbonaceous materials
are very abundant in nature. However, black carbon, which exhibits
86 R. Lenz et al. / Marine Pollution Bulletin 100 (2015) 82–91

Fig. 4. MP of different colour appearance identified visually and by Raman. Left: fibres, right: particles. b./g. = black / grey; t./w. = transparent / white; col. = all colours. Cross marks on top
refer to secondary y-axis (percentages).

spectra of the same kind, is used as a filler in rubbers and to some degree
as a black colourant or dye in other thermoplastics. Given that, black
particles are expected to be generally underestimated in MP counting.
Fig. 7 shows a spectrum of a polymer assigned to ethylene vinyl ace-
tate (EVA), a polymer chemically similar to PE, collected in the English
channel. Due to the incorporation of titanium dioxide (TiO2) nanoparti-
cles simultaneously shows the two peaks at 444 and 610 cm−1. TiO2 is
added to various polymers as a material filler, pigment and UV-blocker.
However, the recognition of the actual polymer type is thereby not im-
peded as its intensity is among that of the polymer and does not overlay
any characteristic peaks. It has to be expected that a standard library
search (one component matching) will result in a lower correlation.
Marine microplastic has been a substrate for the growth of organ-
isms of which residuals can remain on the particles' surface during spec-
tral analysis. Likewise, MP recovered from sediments or animal
intestines may be covered by an organic matrix. High quality Raman
spectra can only be obtained from clean, uncovered surfaces as can be
seen in replicate 3 (Fig. 8). The example shows three measured spots
on a piece of plastic debris collected from Charlottenlund beach,
Denmark. Replicate 1 shows a carbon-like band fully overlaying the
polymer type signal due to clear fouling appearances in this area. For
the cleaner replicate 2 the type can be recognised, although the finger-
print region displays a lifted baseline and the overall spectrum is noisier.
3.3. UV degradation effects on Raman signals
Fig. 9 shows the characteristic peaks of LDPE at 1063 cm−1,
1130 cm− 1, 1296 cm− 1, 1418 cm− 1, 1441 cm−1 and 1463 cm−1 in
the fingerprint region, normalised to the maximum peak. The resulting
Raman spectra after exposure to UV radiation showed a lower intensity
of several characteristic bands (marked with arrows). Changes occur at
1418 cm−1, which is assigned to CH2 wagging in crystalline regions, at

Fig. 5. Spectra acquired from ABS from the same manufacturer in different colours. Reference spectrum from a different product with pale colour. Note the high intensity ranges for yellow
and red caused by colour pigment signal overlays. Collection time 3 s.
 R. Lenz et al. / Marine Pollution Bulletin 100 (2015) 82–91
Fig. 6. Two spectra (blue and black) showing the vibrational band of amorphous carbon as found for many black particles of diverse morphologies, (a) and (b). Image (c) which is of very
similar morphology as (a) displays a PP spectrum (red) instead. Scale bars 20 μm.
1441 cm−1 corresponding to CH2 deformation and at 1463 cm−1 due to
CH2 bending from crystalline and amorphous regions. The peak at
2849 cm−1 with its adjacent minimum also displays a decreasing
trend. We observed these effects in repeated measurements of PE ob-
jects obtained from different sources. The other peaks in the fingerprint
region do not show considerable changes.
The Raman spectrum of PVC after photo-degradation (Fig. 10) shows
the simultaneous decrease of the neighbouring peaks at 693 cm−1 and
637 cm−1, which correspond to the characteristic C\\Cl bonds in the
polymer. The spectra have been normalised to the peak at 2919 cm−1,
which is assigned to C\\H stretching vibrations and considered unaffect-
ed by degradation (Gilbert et al., 2013). The spectrum with highest UV
exposure (Fig. 10f) does not exert this peak, wherefore the relative inten-
sity of the peaks in this spectrum cannot be compared. The spectrum is
characterised by the complete absence of the double peaks and the de
novo appearance of two strong peaks (marked with *, Fig. 10) at
1139 cm−1 and 1540 cm−1, assigned to carbon double bonds (C_C).
PA showed a marginal decrease in the vibrational frequency range at
3306 cm−1 assigned to the amide group in the polymer. PET samples
did not show any consistently traceable spectral changes over the peri-
od of the UV exposure experiment.
Generally we could not observe consistent differences between the
replicates submerged in MilliQ water and seawater throughout the
course of the experiment.
4. Discussion
4.1. Applicability of Raman for MP analysis
Raman microscopy was successfully applied as a technique for
microplastic identification and quantification in the present study.
Fig. 7. Spectrum of a blue microplastic particle (blue, inset with scale bar 100 μm) matching with the EVA reference (red) and two additional peaks due to TiO2 (black).
87
Using a Raman microscope samples can be searched and measured at
the same time resulting in a higher degree of identification certainty
at the cost of approximately double work time required. Because the mi-
croscopic analysis in this study was a precursor to Raman analysis the
numbers of Raman-confirmed MP cannot be larger than the numbers
of visually identified plastics, in contrast to a comparable FTIR study
(Song et al., 2015).
The application of Raman or any other spectroscopic method can as-
sist MP identification as an extension to normal light microscopy. For this
approach the analysis is still visual but the inclusion or exclusion of
doubtful particles can be improved by exemplary spectroscopic analysis.
However, two major drawbacks occur using this procedure. Firstly the
spectral quality will not always allow unambiguous decisions, as marine
MP comprises of a complex mixture of biological, synthetic and inorganic
materials. Secondly, only the number of false-positive identified objects
can be minimised, whereas MP that was not detected by visual analysis
will remain as a unknown false-negative fraction unless it is possible to
measure each particle present in the sample. The latter will, however, be-
come very costly if a large number of samples were to be processed.
The success rate of purely visually compared to Raman analysed par-
ticles decreased with size. For particles b100 μm the success rate falls
below 80%, which makes microscopic analysis an inappropriate method.
The reason for the low success rate is not attributable to visibility alone,
as it would be possible to use a higher magnification. MP of small sizes
are difficult to recognise because decision criteria are based purely on
morphological features which diminish with decreasing size. We show
that with increasing particle size recognition improves because larger
particles exhibit more identification criteria, such as hardness and phys-
ical bending, which become of relevance for particles N1 mm (Song
et al., 2014). However, the size classes investigated in this study were
(except a few fibres) below this threshold.
88 R. Lenz et al. / Marine Pollution Bulletin 100 (2015) 82–91
Fig. 8. Spectra showing the effect of organic matter coating on the polymer matrix, with decreasing effects from rep. 1 to 3. Images to the right show corresponding degree of plastic surface
fouling. Scale bar 20 μm. Red square indicates point of Raman measurement. Collection time 20 s.
Apart from the gain in accuracy of identification results by using a
spectral analysis method, the additional information of the polymer
type is useful in regard to environmental impact or risk assessment.
Polymers differ in sorption capacities and toxicity wherefore it is valu-
able to know their respective distributions (Lee et al., 2014).
4.2. Raman signal confounding factors
Raman spectra of the polymer type can be overlaid by the signal de-
rived from additives, resulting in a considerable modification of the
spectra. These additives are mostly restricted to fillers, pigments and
dyes. Their effects on the Raman signal can be categorised as i) foreign
band overlay, ii) fluorescence and iii) absorbance. Each of these are
discussed in turn below.
There is a chance that MP particles or fibres were not recognised
or disqualified if the overlay of an unknown additive resulted in a
Fig. 9. Comparison of Raman spectra of a photo-exposed LDPE sample (b, c) and an un-
treated control (a). Exposure time is given as an equivalent to midday sun hours. The spec-
tra have been intensity normalised and smoothed by a window width of 24 cm−1.
complete masking of the polymer type. This has to be expected as a
drawback of library matching spectroscopic analysis, as reference librar-
ies cannot cover the whole variety present in the marine samples. In fi-
brous samples obtained from the marine environment it is likely that in
the presence of a very clear dye spectrum, the material is of cellulosic or-
igin (cotton, viscose rayon) due to its own weak response signal (Bart,
2006). Spectra recognised to be only partly overlaid by foreign peaks
were accepted if the underlying polymer type signal was still clearly
present.
Many organic colourants, pigments and dyes strongly fluoresce in
visible light and can thereby hinder spectra acquisition, whereas inor-
ganic pigments are more likely to change the displayed peak pattern
(Fredericks, 2012). Fluorescence is often named as a major factor of
poor Raman signal quality (Fredericks, 2012; Jochem and Lehnert,
2002). The light intensities emitted by fluorescence can be orders of
magnitude stronger than the Raman scattering. Especially measure-
ments of red and yellow coloured items are known to be affected
Fig. 10. Comparison of Raman spectra of a photo-exposed PVC sample (b–f) and an un-
treated control (a). Exposure time is given as an equivalent to midday sun hours. The spec-
tra have been intensity normalised and smoothed by a window width of 24 cm−1. * new
occurrence of peaks.
 R. Lenz et al. / Marine Pollution Bulletin 100 (2015) 82–91
(Jochem and Lehnert, 2002). In our samples red and yellow particles
(Fig. 4, only red shown) had the lowest rates of Raman confirmation
of all colours. The laser wavelength applied in this study was in the vis-
ible blue range (455 nm). Although most authors reported that using a
lower wavelength can mitigate fluorescence signals (Fredericks, 2012),
there is evidence that particularly red dyes used in textiles can have a
fluorescence minimum at blue excitation wavelengths 458 - 488 nm
(Massonnet et al., 2005). The effect of Raman scattering increases with
excitation energy, wherefore short laser wavelength can be used to im-
prove detection of weak Raman active compounds (Wartewig, 2003).
Improvements of detection may also be achieved using techniques
such as Resonance Raman, Surface Enhanced Raman (SERS),
Coherent-Anti-Stokes-Raman (CARS, see also (Cole et al., 2013)) or
Fourier-Transform-Raman (FT-Raman) which can – in specific cases –
substantially increase signal-to-noise ratios, but come at the cost of
more advanced equipment and/or required sample preparation
(Fredericks, 2012). Further investigations have to reveal optimal instru-
mentation and excitation wavelengths for the application of Raman
spectroscopy for marine microplastic analysis.
A quarter of the microplastic data set were assigned plastic-like
polymers giving moderate match scores with PET, however, missing
the characteristic peak at 1616 cm−1. Scanning a number of representa-
tives using the spectroscopy software, Bio Rad-KnowItAll, with a
large library of polymeric compounds and additives, synthetic azo-
compounds appeared among the top scores. However, no single compo-
nent could be attributed to describe this group which hence was includ-
ed as anthropogenic material of unidentified polymeric composition.
Azo-dyes and pigments are commonly used in textiles of cellulosic
and synthetic polymers, and to some extent in other plastic objects as
well. Because the azo-group signal (N_N) is intense in Raman scatter-
ing it partly overlies the actual material type. Other contemplable com-
pounds that could explain some of the present spectral bands were PU
and several polyesters. A minor group of the analysed MP (1.3%)
showed similarities with the above named group in the region around
the dominant peak of 1593 cm−1. In contrast to the afore-mentioned
group no further bands in the fingerprint region were distinguishable
from the background noise. A library match revealed a weak correlation
with a spectrum of a LDPE reference which was overlaid by strong
Raman scattering yellow pigment.
Common polymer fillers such as glass, clay and silica lead to consid-
erable obscuration of spectra in FTIR, however are poorly Raman-active
because of the low polarisability of Si\\O bonds (Bart, 2006). Titanium
dioxide, another filler occasionally observed in our spectra (Fig. 7)
gave additional peaks, which however did not mask the remaining
part of the spectrum.
Black polymers that contain carbon fillers such as carbon black,
amorphous coke or graphite which strongly absorb the light scattered
by the polymer can thus exhibit a carbon-like spectrum. By using a
high laser power it is possible to even burn the sample (Bart, 2006). Par-
ticles originating from road wear (tires) are therefore not discernible to
other natural carbon-rich sources. This can be part of the reason why
black particles were sparsely confirmed via Raman spectroscopy
(Fig. 4). The problem of non-identifiable black particles has been stated
in an earlier study using FTIR (Norén and Naustvoll, 2010).
Other additives are poorly documented for Raman libraries, due to
weak Raman scattering and unsatisfactory reproducibility (Hummel,
2002). Nonetheless, these chemicals can lead to partial masking or a de-
crease of the signal to noise ratio leading to a lower degree of spectral
matching. On the other hand, most additives are not covalently bonded
to the polymer and are leaching out during exposure to the marine con-
ditions such as UV radiation and varying salinity and temperature
(Teuten et al., 2009; Engler, 2012). Regarding the detrimental effect of
organic matter coating (Fig. 8) it is helpful to obtain clean MP surfaces
prior to spectral acquisition and replicate measurements of different
spots on a particle should be conducted. Several studies have applied
acid, base, oxidant or enzymatic digestions steps in sample preparation
89
to remove natural material and coatings (Cole et al., 2014). For S-MP,
however, chemical pre-treatments can easily result in loss of particles
of susceptible polymers even if the bulk polymer can be regarded as re-
sistant to the reagent. In the present study we have applied SDS in the
washing of samples. The surfactant facilitates the dissolution of coatings
from hydrophobic surfaces. SDS is known as a strong Raman scatterer
(Bart, 2006), however, can be removed by thorough rinsing with
water. In the spectral analysis we found no sign of the presence of SDS
in the samples.
4.3. UV degradation effects on Raman signals
Photo-oxidation in polyolefin polymers usually involves reactions
due to free radicals resulting in chain cleavage, cross-linking and the for-
mation of hydroxyl- and carbonyl groups. The formation of these polar
oxidative functional groups can be easily observed via IR measurements.
This is part of the reason why FTIR obtained spectra of marine plastics
often report considerably high levels of impurity, occasionally impeding
accurate spectra acquisition (Norén, 2007; Song et al., 2015). This is less
of a problem with Raman spectroscopy acquired spectra.
The polyolefins PE and PP are most abundant in the surface of the
ocean due to their lower density than seawater (48% in the present
data set, (Enders et al., 2015). Our results on the UV exposure of various
kinds of polymers showed that Raman polymer identification is to a
large extent unaffected of UV-induced weathering of PE and PP. After
1634 h of simulated sunlight in air, both PE and PP showed a slight re-
duction in intensity of characteristic peaks (Fig. 9). Correlation to the
corresponding reference spectra was slightly reduced: for PE by
5.6% ± 2.6% and for PP 8.5% ± 6.4%. The alteration is small and polymer
type recognition was not impeded. Signs of degradations as loss of flex-
ibility and embrittlement were also clearly observable, where especially
the investigated PP samples showed cracks covering the entire polymer
which is in consistency with observations other UV exposure studies
(Andrady, 2011).
PVC degradation is mostly governed by the dehydrochlorination re-
action, which can be chemically, thermally or photochemically induced
(Bowley et al., 1988). In contrast to the above discussed polymers it is
free of hydroxilic or carboxylic group formation. The observed changes
in the investigated PVC spectra with increased UV exposure can be ex-
plained as follows. The exposure to UV radiation initialises the elimina-
tion of hydrochloric acid HCl and the subsequent formation of a C_C
double bond. The loss in chlorine is apparent in the shrinkage of the
carbon-chlorine stretching C-Cl bands as observed in Fig. 10a–e. The
continuation of this, known as the ‘unzipping’ reaction, occurs increas-
ingly fast and leads to the formation of conjugated polyene, which in
the Raman spectra lead to the emergence of the two intense peaks
(Fig. 10f). The vibrational bonds of C_C are easy to detect as they are
very intense Raman scatterers. The intensity of the C_C bands is deter-
mined by the length of the conjugated sequences.
When degradation has reached a certain degree, cross-linking be-
tween the highly reactive polyenes takes place that in turn highly influ-
ences physical and mechanical properties. This then causes a shortening
of the polyenes and consequently to the elimination of the plasticiser
(Bowley et al., 1988). This then leads to typical weathering effects,
such as embrittlement and yellowing, as has been clearly observed
for the investigated PVC sample. Documentation and quantification of
the rapid reduction in mechanical integrity has been studied before
(Al-Dossary et al., 2009).
PVC is, on the one hand, relatively prone to photo-degradation in the
marine environment as the leakage of the large content of additives,
such as photo-stabilisers, is enhanced under high humidity conditions
(Singh and Sharma, 2008). On the other hand, low water temperatures
generally decelerate reactivity. For polymer identification studies via
Raman spectroscopy we recommend to include spectra of degraded
polymer stages to the reference library in order to enhance recognition
possibility. Our results from the degradation test revealed that PVC
90 R. Lenz et al. / Marine Pollution Bulletin 100 (2015) 82–91
Raman spectra can change to a level that makes successful identification
impossible if based on a virgin PVC reference. For PA and PET
degradation-related spectral changes are possible in Raman spectrosco-
py (Neha et al., 2014), although such trends could not be observed to be
strong enough to impede the polymer identification from our data.
It has proven successful to extend the used library by secondarily ob-
tained spectra. We included some of the sample spectra that had highest
matches to the library base after individual evaluation which substan-
tially increased the coverage of chemical variance for the targeted plas-
tic types. Additionally, a useful spectroscopic library for environmental
MP identification should include spectra of exemplary additive groups,
as well as degradation products and intermediate steps of those poly-
mers that are known to exhibit Raman detectable changes under expo-
sure conditions. Library searches should be performed by multi-
component correlations to better identify copolymers and increase
matches in case of signal overlays from additives.
Raman measurements should preferably take place on a non-
Raman-active carrier such as microscopy glass slides to avoid back-
ground signals. In the present study sample material was measured on
two types of vacuum filters, PA and PC. They were deemed suitable
due to their smooth surface and were available in 10 μm pore size,
which was set as the lower size limit of this study. Modern Raman
equipment has a high vertical resolution at the lower micrometre
scale (Sasic and Ozaki, 2011) and scattering effects of underlying or
overlying sample material can be avoided by accurate focussing on the
particles surface. For the development of spectral mapping for automat-
ed sample measurements a non-scattering carrier would be a necessity.
First progress in development of such methods has recently been made
by the utilisation of silicon wafers as MP filter substrate (Käppler et al.,
2015).
4.4. Conclusion and perspectives
Raman spectroscopy provides adequate techniques for the identifi-
cation and chemical analysis of marine microplastic in the lower
micrometre range. Bench-top Raman micro-spectrometer can be used
by the non-expert user and relevant chemical information can be ob-
tained using advanced spectral library software. Raman signals from
photodegraded samples allowed an extended identification for the
most common plastic types in the marine environment. We have
shown, that pure visual identification using morphological criteria led
to a misidentification of a substantial amount of particles (32%) and fi-
bres (25%), which suggests that microplastic studies using visual identi-
fication only involve a high level of uncertainty of the reported
concentrations.
For future research modern Raman microscopes can provide
spectral mapping at high enough speeds to map a whole sample area
in high spatial resolution. In combination with reliable analysis algo-
rithms a manual investigation on individual particle basis could
be made obsolete for counting, polymer type identification and size
determination.
Besides the afore-mentioned mapping and imaging procedures an
alternative approach lies within applications of in situ acquisition of
MP data. This could be realised in continuously measuring flow-
through Raman microspectrometers that can detect MP directly in sea-
water without the need of conserving samples on filters. Scientific
cruises applying such equipment would be able to directly monitor hor-
izontal and vertical profiles of small microplastic concentrations, types
and sizes. Similar techniques have been developed for detection of inor-
ganic nanoparticles in gas flows (Liu et al., 2010), as well as for measur-
ing dissolved organic pollutants in seawater (Kolomijeca, 2013). The
sample volumes that can be ‘scanned’ per unit time will probably be
the most limiting factor in such set-ups. However, it might be overcome
by combining the detection with a preceding step of up-concentration.
Providing these automated, rapid and reliable MP analysis methods is
a key to answer open questions of MP dynamics and their environmen-
tal effects.
Acknowledgements
We thank Eik Ehlert Britsch and Magnus Bohr, which contributed to
the sample collection and Peter Munk for excellent logistics and organi-
sation of the expedition. Thanks to Peter Bøggild from DTU Nanotech for
the opportunity to use the Raman microspectrometer. The research was
funded by DTU Aqua, Carlsberg Foundation (2012-01-0272) and the
Danish Center for Marine Research (2013-02).
References
Al-Dossary, A.K., Gilbert, M., Hitt, D., 2009. Evaluating PVC degradation using UV and
Raman spectroscopies. Adv. Mater. Res. 83-86, 923–930. http://dx.doi.org/10.4028/
www.scientific.net/AMR.83-86.923.
Andrady, A.L., 2003. Plastics and the Environment. John Wiley & Sons, Inc., Hoboken,
New Jersey.
Andrady, A.L., 2011. Microplastics in the marine environment. Mar. Pollut. Bull. 62 (8),
1596–1605. http://dx.doi.org/10.1016/j.marpolbul.2011.05.030.
Avio, C.G., Gorbi, S., Milan, M., Benedetti, M., Fattorini, D., D'Errico, G., Pauletto, M.,
Bargelloni, L., Regoli, F., 2015. Pollutants bioavailability and toxicological risk from
microplastics to marine mussels. Environ. Pollut. 198, 211–222. http://dx.doi.org/
10.1016/j.envpol.2014.12.021.
Bakir, A., Rowland, S.J., Thompson, R.C., 2014. Enhanced desorption of persistent
organic pollutants from microplastics under simulated physiological conditions.
Environ. Pollut. 185, 16–23. http://dx.doi.org/10.1016/j.envpol.2013.10.007.
Barnes, D.K.a., Galgani, F., Thompson, R.C., Barlaz, M., 2009. Accumulation and
fragmentation of plastic debris in global environments. Philos. Trans. R. Soc. Lond.
Ser. B Biol. Sci. 364 (1526), 1985–1998. http://dx.doi.org/10.1098/rstb.2008.0205.
Bart, J.C.J., 2006. Plastics Additives: Advanced Industrial Analysis. IOS Press.
Bhargava, R., Wang, S.-q., Koenig, J.L., 2003. FTIR microspectroscopy of polymeric systems.
Adv. Polym. Sci. 163, 137–191. http://dx.doi.org/10.1007/b11052.
Bonhomme, S., Cuer, a., Delort, a M., Lemaire, J., Sancelme, M., Scott, G., 2003. Environ-
mental biodegradation of polyethylene. Polym. Degrad. Stab. 81 (3), 441–452.
http://dx.doi.org/10.1016/S0141-3910(03)00129-0.
Bowley, H., Gerrard, D., Biggin, I., 1988. The use of laser Raman spectroscopy to study
the degradation of poly(vinyl chloride). Polym. Degrad. Stab. 20 (3-4), 257–269.
http://dx.doi.org/10.1016/0141-3910(88)90072-9.
Browne, M., Galloway, T., Thompson, R., 2007. Microplastic an emerging con-
taminant of potential concern? Integr. Environ. Assess. Manag. 3 (4), 559–566.
Cole, M., Lindeque, P., Halsband, C., Galloway, T.S., 2011. Microplastics as contaminants
in the marine environment: a review. Mar. Pollut. Bull. 62 (12), 2588–2597. http://
dx.doi.org/10.1016/j.marpolbul.2011.09.025.
Cole, M., Lindeque, P., Fileman, E., Halsband, C., Goodhead, R., Moger, J., Galloway, T.S.,
2013. Microplastic ingestion by zooplankton. Environ. Sci. Technol. 47 (12),
6646–6655. http://dx.doi.org/10.1021/es400663f.
Cole, M., Webb, H., Lindeque, P.K., Fileman, E.S., Halsband, C., Galloway, T.S., 2014.
Isolation of microplastics in biota-rich seawater samples and marine organisms.
Sci. Rep. 4, 4528. http://dx.doi.org/10.1038/srep04528.
Cole, M., Lindeque, P., Fileman, E., Halsband, C., Galloway, T.S., 2015. The Impact of Poly-
styrene Microplastics on Feeding, Function and Fecundity in the Marine Copepod
Calanus helgolandicus. Environ. Sci. Technol. http://dx.doi.org/10.1021/es504525u.
Desforges, J.P.W., Galbraith, M., Dangerfield, N., Ross, P.S., 2014. Widespread distribution
of microplastics in subsurface seawater in the NE Pacific Ocean. Mar. Pollut. Bull. 79
(1–2), 94–99. http://dx.doi.org/10.1016/j.marpolbul.2013.12.035.
Edwards, H.G.M., 2005. Modern Raman Spectroscopy—A Practical Approach. vol. 36.
http://dx.doi.org/10.1002/jrs.1320.
Enders, K., Lenz, R., Stedmon, C.A., Nielsen, T.G., 2015. Abundance, Size and Polymer
Composition of Marine Microplastics Larger 10 Micrometre In The Atlantic Ocean
And Their Modelled Vertical Distribution. Mar. Pollut. Bull. http://dx.doi.org/10.
1016/j.marpolbul.2015.09.027 (in preparation).
Engler, R.E., 2012. The complex interaction between marine debris and toxic chemicals in
the ocean. Environ. Sci. Technol. 46, 12302–12315. http://dx.doi.org/10.1021/
es3027105.
Eriksen, M., Maximenko, N., Thiel, M., Cummins, A., Lattin, G., Wilson, S., Hafner, J., Zellers,
A., Rifman, S., 2013. Plastic pollution in the South Pacific subtropical gyre. Mar. Pollut.
Bull. 68 (1-2), 71–76. http://dx.doi.org/10.1016/j.marpolbul.2012.12.021.
Fredericks, P., 2012. Forensic analysis of fibres by vibrational spectroscopy. In: Chalmers,
J.M., Edwards, H.G.M., Hargreaves, M.D. (Eds.), Infrared and Raman Spectroscopy in
Forensic. Wiley, pp. 153–169 (Ch. 4.4,).
Fries, E., Dekiff, J.H., Willmeyer, J., Nuelle, M.-T., Ebert, M., Remy, D., 2013. Identification
of polymer types and additives in marine microplastic particles using pyrolysis-GC/
MS and scanning electron microscopy. Environ. Sci.: Processes Impacts 15 (10),
1949–1956. http://dx.doi.org/10.1039/c3em00214d.
Gilbert, M., Ho, K.C., Hitt, D.J., Vrsaljko, D., 2013. Assessment of PVC stabilisation
using hydrotalcites — Raman spectroscopy and other techniques. Polym. Degrad.
Stab. 98 (8), 1537–1547. http://dx.doi.org/10.1016/j.polymdegradstab.2013.04.005.
Goldstein, M.C., 2012. Abundance And Ecological Implications Of Microplastic Debris In
The North Pacific Subtropical Gyre (Ph.D. thesis) University of California, San Diego.
 R. Lenz et al. / Marine Pollution Bulletin 100 (2015) 82–91
Graca, B., BeLdowska, M., Wrzesień, P., Zgrundo, A., 2014. Styrofoam debris as a potential
carrier of mercury within ecosystems. Environ. Sci. Pollut. Res. Int. 21 (3), 2263–2271.
http://dx.doi.org/10.1007/s11356-013-2153-4.
Hidalgo-Ruz, V., Gutow, L., Thompson, R.C., Thiel, M., 2012. Microplastics in the marine
environment: a review of the methods used for identification and quantification.
Environ. Sci. Technol. 46 (6), 3060–3075. http://dx.doi.org/10.1021/es2031505.
Hummel, D.O., 2002. Atlas of Plastics Additives Analysis by Spectrometric Methods.
Springer-Verlag Berlin and Heidelberg GmbH & Co. K.
Imhof, H.K., Schmid, J., Niessner, R., Ivleva, N.P., Laforsch, C., 2012. A novel, highly efficient
method for the separation and quantification of plastic particles in sediments of
aquatic. Limnol. Oceanogr. Methods 10, 524–537. http://dx.doi.org/10.4319/lom.
2012.10.524.
Ivar do Sul, J.a., Costa, M.F., 2014. The present and future of microplastic pollution in the
marine environment. Environ. Pollut. 185, 352–364. http://dx.doi.org/10.1016/j.
envpol.2013.10.036.
Jochem, G., Lehnert, R., 2002. On the potential of Raman microscopy for the forensic anal-
ysis of coloured textile fibres. Sci. Justice 42 (4), 215–221. http://dx.doi.org/10.1016/
S1355-0306(02)71831-5.
Käppler, A., Windrich, F., Löder, M.G.J., Malanin, M., Fischer, D., Labrenz, M., Eichhorn, K.-J.,
Voit, B., 2015. Identification of microplastics by FTIR and Raman microscopy: a novel
silicon filter substrate opens the important spectral range below 1300 cm−1 for FTIR
transmission measurements. Anal. Bioanal. Chem. 6791–6801 http://dx.doi.org/10.
1007/s00216-015-8850-8.
Koenig, J.L., 2001. Infrared and Raman Spectroscopy of Polymers. vol. 12. Smithers Rapra
Press.
Kolomijeca, A., 2013. An Autonomous Sea Going Raman/SERS Instrument For In Situ De-
tection Of Chemicals In Sea Water (Ph.D. thesis) Technische Universität Berlin.
Lagaron, J.M., Dixon, N.M., Reed, W., Pastor, J.M., Kip, B.J., 1999. Morphological character-
isation of the crystalline structure of cold-drawn HDPE used as a model material
for the environmental stress cracking (ESC) phenomenon. Polymer 40 (10),
2569–2586. http://dx.doi.org/10.1016/S0032-3861(98)00500-X.
Lambert, J.B., 1987. Peak interpretation lists.pdf. Introduction to Organic Spectroscopy.
Lattin, G.L., Moore, C.J., Zellers, a.F., Moore, S.L., Weisberg, S.B., 2004. A comparison of
neustonic plastic and zooplankton at different depths near the southern California
shore. Mar. Pollut. Bull. 49 (4), 291–294. http://dx.doi.org/10.1016/j.marpolbul.
2004.01.020.
Lee, K.-w., Shim, W.J., Kwon, O.Y., Kang, J.-h., 2013. Size-dependent effects of micro poly-
styrene particles in the marine copepod Tigriopus japonicus. Environ. Sci. Technol. 47,
11278–11283. http://dx.doi.org/10.1021/es401932b (August).
Lee, H., Shim, W.J., Kwon, J.-H.H., 2014. Sorption capacity of plastic debris for hydrophobic
organic chemicals. Sci. Total Environ. 470-471, 1545–1552. http://dx.doi.org/10.1016/
j.scitotenv.2013.08.023.
Liu, X., Smith, M.E., Tse, S.D., 2010. In situ Raman characterization of nanoparticle aerosols
during flame synthesis. Appl. Phys. B Lasers Opt. 100 (3), 643–653. http://dx.doi.org/
10.1007/s00340-010-4091-x.
Magnusson, K., Wahlberg, C., 2014. Mikroskopiska skräppartiklar i vatten frå n
avloppsreningsverk. Tech. repNR B 2208. Svenska Miljöinstitutet.
Massonnet, G., Buzzini, P., Jochem, G., Stauber, M., Coyle, T., Roux, C., Thomas, J.,
Leijenhorst, H., Van Zanten, Z., Wiggins, K., Russell, C., Chabli, S., Rosengarten, A.,
2005. Evaluation of Raman spectroscopy for the analysis of colored fibers: a
collaborative study. J. Forensic Sci. 50 (5), 1028–1038. http://dx.doi.org/10.1520/
JFS2004532.
Moore, C.J., 2008. Synthetic polymers in the marine environment: a rapidly increasing,
long-term threat. Environ. Res. 108 (2), 131–139. http://dx.doi.org/10.1016/j.envres.
2008.07.025.
Morét-Ferguson, S., Law, K.L., Proskurowski, G., Murphy, E.K., Peacock, E.E., Reddy, C.M.,
2010. The size, mass, and composition of plastic debris in the western North Atlantic
91
Ocean. Mar. Pollut. Bull. 60 (10), 1873–1878. http://dx.doi.org/10.1016/j.marpolbul.
2010.07.020.
Neha, S., Tripathi, B.K., Shrivastava, A.K., Chauhan, R.S., 2014. Laser Raman Studies Of
Fibrous Polymer — A Review. 5 (2), 128–139.
Norén, F., 2007. Small plastic particles in Coastal Swedish waters. Tech. Rep. 0. KIMO.
Norén, F., Naustvoll, L.-j., 2010. Survey of Microscopic Anthropogenic Particles in Skager-
rak. Tech. Rep. Institute of Marine Research, Flø devigen November.
Norén, F., Naustvoll, L.-J., 2011. Pilot study: survey of microscopic anthropogenic
particles in Skagerrak. Tech. RepKlima- og Forurensningsdirektoratet: TA2779/2011.
N-research and Havforskningsinstituttet, Lysekil, Sweden.
Norén, F., Norén, K., Magnusson, K., 2014. Marint mikroskopiskt skräp Undersökning
längs svenska västkusten. Tech. rep.
Rocha-Santos, T., Duarte, A.C., 2014. A critical overview of the analytical approaches
to the occurrence, the fate and the behavior of microplastics in the environment.
TrAC Trends Anal. Chem. 65, 47–53. http://dx.doi.org/10.1016/j.trac.2014.10.011.
Sasic, S., Ozaki, Y., 2011. Raman, Infrared, and Near-Infrared Chemical Imaging.
Wiley.
Setälä, O., Fleming-Lehtinen, V., Lehtiniemi, M., 2014. Ingestion and transfer of
microplastics in the planktonic food web. Environ. Pollut. 185, 77–83. http://dx.doi.
org/10.1016/j.envpol.2013.10.013.
Singh, B., Sharma, N., 2008. Mechanistic implications of plastic degradation. Polym.
Degrad. Stab. 93 (3), 561–584. http://dx.doi.org/10.1016/j.polymdegradstab.2007.
11.008.
Socrates, G., 2004. Infrared and Raman Characteristic Group Frequencies.
Song, Y.K., Hong, S.H., Jang, M., Kang, J.-H., Kwon, O.Y., Han, G.M., Shim, W.J., 2014. Large
accumulation of micro-sized synthetic polymer particles in the sea surface
microlayer. Environ. Sci. Technol. 48, 9014–9021. http://dx.doi.org/10.1021/
es501757s.
Song, Y.K., Hong, S.H., Jang, M., Han, G.M., Rani, M., Lee, J., Shim, W.J., 2015. A
comparison of microscopic and spectroscopic identification methods for analysis of
microplastics in environmental samples. Mar. Pollut. Bull. 1–8 http://dx.doi.org/10.
1016/j.marpolbul.2015.01.015.
Teuten, E.L., Saquing, J.M., Knappe, D.R.U., Barlaz, M.a., Jonsson, S., Björn, A., Rowland, S.J.,
Thompson, R.C., Galloway, T.S., Yamashita, R., Ochi, D., Watanuki, Y., Moore, C., Viet,
P.H., Tana, T.S., Prudente, M., Boonyatumanond, R., Zakaria, M.P., Akkhavong, K.,
Ogata, Y., Hirai, H., Iwasa, S., Mizukawa, K., Hagino, Y., Imamura, A., Saha, M.,
Takada, H., 2009. Transport and release of chemicals from plastics to the environment
and to wildlife. Philos. Trans. R. Soc. Lond. Ser. B Biol. Sci. 364 (1526), 2027–2045.
http://dx.doi.org/10.1098/rstb.2008.0284.
Vianello, A., Boldrin, A., Guerriero, P., Moschino, V., Rella, R., Sturaro, A., Da Ros, L., 2013.
Microplastic particles in sediments of Lagoon of Venice, Italy: first observations on oc-
currence, spatial patterns and identification. Estuar. Coast. Shelf Sci. 130, 54–61.
http://dx.doi.org/10.1016/j.ecss.2013.03.022.
Wartewig, S., 2003. IR and Raman Spectroscopy — Fundamental Processing. Wiley-VCH
Verlag GmbH & Co. KGaA, Weinheim, FRG http://dx.doi.org/10.1002/3527601635.
Wegner, A., Besseling, E., Foekema, E.M., Kamermans, P., Koelmans, a.a., 2012. Effects of
nanopolystyrene on the feeding behavior of the blue mussel (Mytilus edulis L.). Envi-
ron. Toxicol. Chem. 31 (11), 2490–2497. http://dx.doi.org/10.1002/etc.1984.
Wright, S.L., Thompson, R.C., Galloway, T.S., 2013. The physical impacts of microplastics
on marine organisms: a review. Environ. Pollut. 178, 483–492. http://dx.doi.org/10.
1016/j.envpol.2013.02.031.
Zettler, E.R., Mincer, T.J., Amaral-Zettler, L.a., 2013. Life in the “plastisphere”: microbial
communities on plastic marine debris. Environ. Sci. Technol. 47 (13), 7137–7146.
http://dx.doi.org/10.1021/es401288x.
Zheng, Y., Yanful, E.K., Bassi, A.S., 2005. A review of plastic waste biodegradation. Crit. Rev.
Biotechnol. 25 (4), 243–250. http://dx.doi.org/10.1080/07388550500346359.