Science of the Total Environment 728 (2020) 138449

Contents lists available at ScienceDirect

Science of the Total Environment

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

In situ surface-enhanced Raman spectroscopy for detecting microplastics

and nanoplastics in aquatic environments
Lulu Lv a,b, Lei He a, Shiqi Jiang a,b, Jinjun Chen c, Chunxia Zhou b,d,e, Junhao Qu e, Yuqin Lu a, Pengzhi Hong b,d,e,
Shengli Sun a,⁎, Chengyong Li a,b,d,⁎⁎
a
School of Chemistry and Environment, Guangdong Ocean University, Zhanjiang 524088, China
b
Shenzhen Institute of Guangdong Ocean University, Shenzhen 518108, China
c
College of Coastal Agricultural Sciences, Guangdong Ocean University, Zhanjiang 524088, China
d
Southern Marine Science and Engineering Guangdong Laboratory, Zhanjiang 524088, China
e
College of Food Science and Technology, Guangdong Ocean University, Zhanjiang 524088, China

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• A method for detecting micro/

nanoplastics in water environment was
developed.
• Plastic particles in the size range from
nanometers to microns can be detected.
• Effective identification of different types
of plastic particles.
• Capable of detecting 100 nm plastic par-
ticles down to 40 μg/mL.

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

Article history: The detection of microplastics and nanoplastics in the environment, especially plastic particles in aquatic envi-
Received 25 January 2020 ronments in situ, still faces challenges due to the limitations of current methods, instruments and size of plastic
Received in revised form 17 March 2020 particles. This paper evaluates the potential of surface-enhanced Raman spectroscopy for the analysis of
Accepted 2 April 2020
microplastics and nanoplastics. The condition of different tests including the volume ratio of sample to silver col-
Available online 20 April 2020
loid, the concentrations of NaCl, and the concentrations of the samples, are assessed for the study of microplastics
Editor: Damia Barcelo and nanoplastics (polystyrene (PS), polyethylene (PE) and polypropylene (PP)) in pure water and seawater. A
method based on SERS, that uses silver colloid as the active substrate, is developed for the qualitative analysis
Keywords: of microplastics and nanoplastics in aquatic environments. The particle sizes of microplastics and nanoplastics in-
Microplastics clude 100 nm, 500 nm and 10 μm. The Raman signals of microplastics and nanoplastics in pure water and seawa-
Nanoplastics ter both show good enhancement efficiency. The optimal enhancement factor is 4 × 104. The SERS-based
Pollutant detection method overcomes the limitations of microplastics and nanoplastics in liquids and can detect
Qualitative analysis 100 nm plastics down to 40 μg/mL. It provides more possibility for the rapid detection of microplastics and
nanoplastics in aquatic environments in the future.
© 2020 Published by Elsevier B.V.

⁎ Corresponding author.
⁎⁎ Correspondence to: C. Li, School of Chemistry and Environment, Guangdong Ocean University, Zhanjiang 524088, China.
E-mail addresses: sunsl@gdou.edu.cn (S. Sun), cyli@gdou.edu.cn (C. Li).

https://doi.org/10.1016/j.scitotenv.2020.138449
0048-9697/© 2020 Published by Elsevier B.V.
2 L. Lv et al. / Science of the Total Environment 728 (2020) 138449
1. Introduction
The mass production of plastics has brought convenience to human
daily life (Lodge, 2017). However, the emergence of plastic litter has
brought global threat (Barnes et al., 2009; Browne et al., 2011). Plastic
fragments produced in industrial production, synthetic fibers in cloth-
ing and microbeads in daily chemicals are the main sources of plastic lit-
ter (Cole and Galloway, 2015; Eriksen et al., 2013; Fendall and Sewell,
2009; Geyer et al., 2017). An increasing number of studies have reported
that plastic litter widely exists in almost all habitats worldwide (Eerkes-
Medrano et al., 2015; Eriksen et al., 2014; Law et al., 2010; Law et al.,
2014; Mani et al., 2015; Mintenig et al., 2018; Schlining et al., 2013).
Microplastics are particles smaller than 5 mm in size, which are formed
by the degradation of larger plastic fragments through physical and bi-
ological effects (Gewert et al., 2015; Harshvardhan and Jha, 2013). Al-
ternatively, these particle sizes are produced intentionally.
Microplastics have attracted widespread attention, because they are
easily ingested by organisms and may pose a potential threat to more
organisms and even human health through the food chains (Farrell
and Nelson, 2013; Nam Ngoc et al., 2018; Revel et al., 2018; Setala
et al., 2014; Wright and Kelly, 2017). However, numerous studies
have investigated plastic particles with N20 μm in size (Cabernard
et al., 2018; Eo et al., 2018; Erni-Cassola et al., 2017), while nanoplastics
have rarely been reported (Gigault et al., 2018). The discovery of
nanoplastics in facial scrubs (Hernandez et al., 2017) and ocean surface
water (Ter Halle et al., 2017) has attracted great attention. There is no
uniform standard for this definition so far. Some researchers have de-
fined nanoplastics as particles smaller than 1 μm (da Costa et al.,
2016; Lambert and Wagner, 2016; Ter Halle et al., 2017). Others believe
that plastic particles with at least two dimensions between 1 and
100 nm are defined as nanoplastics (Klaine et al., 2012; Klaine et al.,
2008; Koelmans et al., 2015a; Peiponen et al., 2019).
At present, there are various detection techniques for the analysis of
microplastics, including Pyrolysis gas chromatography–mass spectrom-
etry (Pyrolysis-GC/MS) (Dekiff et al., 2014; Duemichen et al., 2017; Fries
et al., 2013; Kaeppler et al., 2018), Fourier transform infrared spectros-
copy (FTIR) (Huppertsberg and Knepper, 2018; Ngoc Nam et al., 2017;
Suaria et al., 2016) and Raman spectroscopy (Araujo et al., 2018; Lenz
et al., 2015; Shim et al., 2017; Wang and Wang, 2018). These techniques
enable the qualitative and quantitative analysis of microplastics in the
environment. They play a very important role in the detection of
microplastics. However, pyrolysis-GC/MS can only analyze a small num-
ber of samples in each run, only 0.5 mg, and it is not suitable for analyz-
ing complex environmental samples (Duemichen et al., 2017). For FTIR
and Raman spectroscopy, their common limitation is the inability to de-
tect small-sized plastic particles (b1 μm), which has led to a lack of
nanoplastics data (like quantity, chemical properties) in the
environment.
Especially for the detection of microplastics and nanoplastics in
aquatic environments, there are still challenges. Currently, transmission
electron microscopy (TEM) (Gigault et al., 2016) and scanning electron
microscopy (SEM) (Koelmans et al., 2015b) are commonly used to char-
acterize nanoplastics. However, they cannot provide chemical informa-
tion. In the newly published report (Schirinzi et al., 2019), polystyrene
(PS) microplastics and nanoplastics in natural water were quantita-
tively analyzed by liquid chromatography coupled to high-resolution
mass spectrometry (LC-HRMS). The method has high sensitivity and
low detection limits, allowing the analysis in real environmental sam-
ples. However, there are some limitations: (1) It is only applicable to
the detection of PS; (2) The pre-treatment of the sample is cumbersome
and the operation process is complicated.
Surface-enhanced Raman spectroscopy (SERS) is broadly used in
medical (Vo-Dinh et al., 2002; Xie and Schlücker, 2013), food safety
(Feng et al., 2019; He et al., 2011) and environmental monitoring
(Feng et al., 2016; Li et al., 2016; Ma et al., 2013; Xu et al., 2015) due
to its high sensitivity and unique molecular specificity. Generally, the
Raman signal of the sample itself is very weak, but it can be greatly en-
hanced when the sample molecules are adsorbed on the corrugated
metal surface (usually gold or silver nanoparticles). Based on this prin-
ciple, we believe that the detection of microplastics and nanoplastics
with low-concentration aqueous solutions and a smaller size can be
achieved by SERS technology. Metal colloid (Mecker-Pogue et al.,
2012), gold nanocolumn array (Kim et al., 2012) and silver nanoparticle
modified silver nanowires (Chen and Liu, 2012) are used as SERS active
substrates. Among the above active substrates (Mecker-Pogue et al.,
2012; Kim et al., 2012; Chen and Liu, 2012), metal colloids are employed
as substrates for SERS detection because of the simplest preparation and
better SERS activity. Furthermore, some studies have demonstrated that
the addition of salts and acids to the silver colloid would cause the ag-
gregation of the nanoparticles, which can generate a rich SERS hot
spot and enhance the Raman signal (Lin et al., 2014).
In this work, we developed a qualitative analytical method that can
determine the chemical composition of microplastics and nanoplastics
in aqueous solutions. Different plastic types (polystyrene, polyethylene,
and polypropylene) and different plastic sizes (100 nm, 500 nm, 10 μm)
were applied for our experiments. The analytical method is based on
surface-enhanced Raman spectroscopy techniques to test the Raman
signals of microplastics and nanoplastics in pure water and seawater.
Silver colloid was chosen as the active substrate for SERS, and sodium
chloride (NaCl) was employed as an aggregating agent for silver colloid.
The enhanced efficiency of plastic particles with different sizes and
types were investigated by changing the volume ratio of sample to silver
colloid, the concentration of NaCl and the concentration of samples. Fur-
thermore, the microstructure and particle size of silver colloid and
nanoplastics were characterized by transmission electron microscopy
(TEM) and dynamic light scattering (DLS). The results of this study indi-
cated that SERS techniques can be a good analytical approach for the de-
tection of microplastics and nanoplastics in aquatic environments.
2. General methods
2.1. Preparation of microplastics and nanoplastics
The plastic particles of 100 nm and 500 nm PS spheres were synthe-
sized using the method reported by Nandiyanto (Nandiyanto et al.,
2012). First, a round bottom flask containing ultrapure water was
placed in a reactor system with vigorous stirring (600 rpm) and heated
at a specific temperature under a nitrogen atmosphere. The heating pro-
cess was maintained for 30 min to remove oxygen from the solution.
Styrene monomer (ratio of styrene and aqueous solution, 100 nm PS,
v:v = 1:100; 500 nm PS, v:v = 1:9) was added to the reactor for
10 min to ensure that styrene was uniformly dispersed in aqueous solu-
tion. Subsequently, 2,2-azobis (isobutyramidine) dihydrochloride
(AIBA) (100 nm, 10 mg; 500 nm, 0.3 g) previously dissolved in water
was added to the mixture of styrene-water solution. The mixture was
reacted at this temperature (synthetic 100 nm PS spheres at 90 °C and
500 nm PS spheres at 55 °C) for 10 h under nitrogen. A total of the
200 mL of aqueous solution was added. After cooling the reacted solu-
tion to room temperature, excess AIBA and styrene monomer were
washed away with absolute ethanol. The PS sphere of 500 nm was
washed by filtration (over 0.22 μm of filter membrane) and the
100 nm PS sphere was washed by dialysis method. Finally, it was
dried at 50 °C to obtain a solid powder sample. To avoid laboratory con-
tamination, laboratory coats were worn during all procedures, glass-
ware were washed with acetone, other experimental equipment were
thoroughly rinsed with ultrapure water.
PE and PP plastic powder with a size of about 10 μm were purchased
from Yineng Plastic Materials Co., Ltd. (Dongguan, China). They were
separately dispersed in pure water and sea water (50 mg in 10 mL).
The seawater was collected from the surface seawater of Zhanjiang
Bay. Since both microplastics were difficult to disperse in water,
 L. Lv et al. / Science of the Total Environment 728 (2020) 138449
100 μL of surfactant (v/v = 1:100) was added to uniformly disperse the
microplastics.
2.2. Preparation of silver colloid
Silver colloid was selected as the SERS active substrate, which was
prepared by chemical reduction method (Lee and Meisel, 1982). Briefly,
100 mL of a 0.18 g/L solution of AgNO3 was placed in a 250 mL clean
round bottom flask for heating. After the solution boiled, 2.0 mL of 1%
trisodium citrate was quickly added and reacted for 1 h. Here sodium
citrate acted as a reduction and prevented the aggregation of formed
nanoparticles. The solution was naturally cooled to room temperature
and filtered through a 0.22 μm filter membrane to obtain homogeneous
silver nanoparticles. Then it was stored in the dark at 4 °C.
2.3. SERS measurements
Raman spectra were collected using a portable Raman spectrometer
(Ocean Optics, SR-510 Pro) with a 785 nm excitation light. The spectral
measurements were conducted with a 5 s exposure time and laser
power of 105 mW. Each acquisition was repeated 3 times or more to cal-
culate the average value. The spectrometer can provide Raman spectra
in the range of 200 to 3000 cm−1 with a spectral resolution of 5 cm−1.
Accuman software was used for spectra data collection and analysis.
All Raman spectra represented in this study were baseline corrected.
3. Results and discussion
3.1. Detection of microplastics and nanoplastics in pure water
As a liquid SERS substrate, silver colloid can be mixed with a sample
solution in any proportion to produce different enhancement efficien-
cies. The optimal SERS signal can be achieved only when the sample
and silver nanoparticles are maintained at the proper ratio. In our ex-
periments, 100 nm PS and 500 nm PS spheres were respectively
mixed with silver colloid in different ratios to analyze their SERS signals.
The concentration of PS spheres in the aqueous solution was 0.4 mg/mL
and the concentration of NaCl was 1 mol/L. The SERS signals of 100 nm
and 500 nm PS spheres at the characteristic peak of 1000 cm−1 are
shown in Fig. 1a. It can be observed that the SERS signal of both reaches
the highest when the volume ratio of the PS sphere aqueous solution to
the silver colloid is 1:1. Here, the SERS intensity of the 100 nm PS sphere
is significantly lower than that of the 500 nm PS sphere, which is mainly
due to the lower enhancement efficiency of the 100 nm PS sphere at a
NaCl concentration of 1 mol/L.
Fig. 1b shows the SERS signals of 100 nm and 500 nm PS spheres at
different NaCl concentrations. NaCl solutions was used with different
concentrations of 0, 0.1, 0.25, 0.5, 0.75, 1, 2, 3 mol/L. The results indi-
cated that the 100 nm and 500 nm PS spheres have the optimal SERS
signals when the NaCl concentration was 0.25 mol/L. We found that
the SERS signal of the 100 nm PS sphere decreased rapidly in the NaCl
concentration range of 0.25–3 mol/L. This may be because the size of
the 100 nm PS sphere is similar to that of the silver nanoparticles.
When the salt concentration increases in the solution, the silver nano-
particles are heavily aggregated and cannot be adsorbed on the surface
of the 100 nm PS sphere.
Based on the above results, the final volume ratio of sample solution
to silver colloid was 1:1 and NaCl with a final concentration of 0.25 mol/
L in sample was applied as aggregating agent. we tested the SERS inten-
sity at different concentrations of the sample. All samples were mea-
sured three times to average. The concentration of PS sphere was set
to 0, 0.04, 0.08, 0.16, 0.2, 0.4, 0.6, 0.8, 1.2, 1.6 mg/mL. As depicted in
Fig. 1c-d, the SERS signal of 100 nm PS sphere increases with increasing
concentration in the range of 0–0.8 mg/mL. The optimal enhancement
factor for a 100 nm PS sphere is calculated as 5 × 102. The enhancement
factor is not high for two reasons: (1) The 100 nm PS sphere exists in the
3
form of solid powder in aqueous solution, and its surface enhancement
signals must be far less than the surface enhancement of molecules in
solution; (2) Although the Raman signal of the 100 nm PS sphere of
the aqueous solution is very low, it also affects the calculation of the en-
hancement factor. The enhancement factor is calculated according to
the formula provided in the reference (Feng et al., 2019).
The SERS signal of the 500 nm PS sphere is the highest at a concen-
tration of 0.08 mg/mL and the enhancement factor at this time is
4 × 104. The reason why the enhancement factor of 500 nm PS sphere
is higher than that of 100 nm PS sphere is that the Raman signal of
500 nm PS sphere in aqueous solution is very weak or not even at
high concentrations. As presented in Fig. 1e-f, the SERS intensity of the
500 nm PS sphere gradually decreased with increasing concentration
in the range of 0.08–1.6 mg/mL, which may be caused by the aggrega-
tion of excessive silver nanoparticles adsorbed on the surface of
samples.
The SERS signals of PE and PP microplastics were recorded. The
number of acquisitions was set to 10 times, once every 60 s. Due to
the large size, it takes time for the silver aggregates to adhere on the sur-
face of microplastics to achieve signal enhancement. The Raman signals
of PE and PP are shown in Fig. S1. It can be seen that the Raman signal of
PE is enhanced from the beginning and does not change much in
10 min. The Raman signal of PP increases with time in the range of
10 min. Besides, we have found that the enhanced signal of
microplastics is not as strong as nanoplastics. This may be the silver
nanoparticles generated fewer hot spots on the surface of the
microplastics due to the large size of the microplastics.
3.2. Detection of microplastics and nanoplastics in seawater
There is more difficult for the detection of microplastics and
nanoplastics in seawater. This may be because seawater itself has a
strong background (Fig. S2) and there may be various impurities, such
as microorganisms and minerals. We have performed experiments on
100 nm and 500 nm PS spheres in seawater. Firstly, 100 nm PS and
500 nm PS sphere were respectively mixed with silver colloid in differ-
ent ratios to analyze their SERS signals. The concentration of PS sphere
in the aqueous solution was 0.4 mg/mL and the concentration of NaCl
was 1 mol/L. It can be seen from Fig. 2a that the SERS signal of both
reaches the highest when the volume ratio of the PS sphere aqueous so-
lution to the silver colloid is 1:1. Secondly, the SERS intensity of 100 nm
and 500 nm PS sphere under different NaCl concentration conditions
were also tested (Fig. 2b). It can be observed that the optimal NaCl con-
centration for enhancing the Raman signal of 100 nm PS and 500 nm PS
spheres in seawater is 0.25 mol/L, which is the same as pure water. The
reason may be that the salt concentration in the seawater is low without
affecting the concentration of the final solution.
Finally, the samples with different concentrations were dispersed in
seawater to measure Raman signals. The final volume ratio of the sam-
ple solution to silver colloid was 1:1 and the NaCl concentration was
0.25 mol/L. As depicted in Fig. 2c-d, the SERS intensity of the 100 nm
PS sphere increases with increasing concentration. The optimal en-
hancement factor for a 100 nm PS sphere is calculated as 7 × 102 and
the results are similar to those of a 100 nm PS sphere in pure water.
The 500 nm PS sphere in seawater has the strongest SERS signal at
0.08 mg/mL of concentration (Fig. 2e-f), and its enhancement factor is
calculated as 1.1 × 104. The SERS signal of PE in seawater is also tested.
The results are shown in Fig. S3, the Raman signal of PE is enhanced after
adding the silver colloid compared to the conventional Raman signal.
But the enhanced signal is not very high for the same reason as in
pure water.
From the above results, we find that the SERS signal of 100 nm and
500 nm PS sphere in seawater have similar trends to those in pure
water. For example, the SERS signal of 100 nm PS sphere in pure
water or seawater gradually increases with increasing concentration.
For 500 nm PS sphere in pure water or seawater, the SERS signal
4 L. Lv et al. / Science of the Total Environment 728 (2020) 138449

Fig. 1. SERS intensity of 100 nm PS spheres and 500 nm PS spheres in pure water at 1000 cm−1 characteristic peaks. (a) The effect of different volume ratios of sample and silver colloid on
the SERS intensity of PS spheres. (b) The influence of different NaCl concentrations on the SERS signal of PS spheres. (c) The SERS spectra of 100 nm PS sphere at different concentrations.
(d) The SERS intensity of 100 nm PS sphere at different concentrations. (e) The SERS spectra of 500 nm PS sphere at different concentrations. (f) The SERS intensity of 500 nm PS sphere at
different concentrations.

increases first and then decreases with increasing PS concentration. An-
other interesting result is that 500 nm PS sphere both in pure water and
seawater has the strongest SERS signal at 0.08 mg/mL. Therefore, we
consider that the SERS signal of the PS sphere does not seem to be dis-
turbed by the seawater environment. The possible reason is that the col-
lected seawater itself is relatively pure, and the impurities in the
seawater have been removed after filtering through a 0.22 μm filter
membrane.
3.3. Characterization analysis
The optical absorption characteristics of the silver colloid were mea-
sured using an ultraviolet-visible spectrum. As can be seen from Fig. S4
a, the maximum absorption peak of the silver colloid is 407 nm, and its
peak shape is narrow and no other peaks appear. It is preliminarily be-
lieved that the prepared silver colloid has a uniform particle size distri-
bution. NaCl solution was used in our experiments because it allowed
 L. Lv et al. / Science of the Total Environment 728 (2020) 138449 5

Fig. 2. SERS intensity of 100 nm PS spheres and 500 nm PS spheres in seawater at 1000 cm−1 characteristic peaks. (a) The effect of different volume ratios of sample and silver colloid on the
SERS intensity of PS spheres. (b) The influence of different NaCl concentrations on the SERS signal of PS spheres. (c) The SERS spectra of 100 nm PS sphere at different concentrations.
(d) The SERS intensity of 100 nm PS sphere at different concentrations. (e) The SERS spectra of 500 nm PS sphere at different concentrations. (f) The SERS intensity of 500 nm PS
sphere at different concentrations.

the silver nanoparticles to aggregate. Fig. S4 b shows the ultraviolet-
visible spectrum of silver nanoparticles after the addition of NaCl solu-
tion. It can be seen that there is an obvious surface plasmon resonance
at 795 nm which indicates the formation of the aggregated silver nano-
particles. The aggregation state of the silver nanoparticles was deter-
mined by characterizing the particle size in the solution before and
after the aggregation. Transmission electron microscopy (TEM, Tecnai
G2F20S) was used to characterize silver nanoparticles and silver aggre-
gates. The results are shown in Fig. 3a and b, most of the silver
nanoparticles are spherical in shape and dispersed. After the addition
of NaCl solution, the potential balance of the colloid is destroyed,
resulting in the aggregation of silver nanoparticles. To study the particle
size distribution of silver nanoparticles before and after aggregation,
they were characterized by Dynamic Light Scattering (DLS). As shown
in the subplot in Fig. 3a and b, the particle diameter of the silver nano-
particles prepared is mainly distributed in the range of 40 nm to
70 nm, and the average diameter is 56 ± 0.82 nm. The size of the silver
nanoparticle aggregates is mainly distributed between 1500 nm and
6 L. Lv et al. / Science of the Total Environment 728 (2020) 138449

Fig. 3. The characterization images of silver nanoplastics and PS spheres. (a) TEM and DLS image of a dispersed silver nanoparticles. (b) TEM and DLS image of aggregated silver
nanoparticles. (c) TEM and DLS image of a dispersed 100 nm PS spheres. (d) TEM and DLS image of 100 nm PS spheres after adding silver colloid. (e) TEM and DLS image of a
dispersed 500 nm PS spheres. (f) TEM and DLS image of 500 nm PS spheres after adding silver colloid.

2000 nm, which is nearly 20 times larger than that before adding coag-
ulant. This indicates that the silver colloid has good SERS enhancement
efficiency. Furthermore, we also characterized the PS sphere solution
before and after the addition of silver colloid by TEM and DLS, respec-
tively. As observed in Fig. 3c–f, the diameters of the synthesized PS
sphere are about 100 nm and 500 nm, respectively. After adding the sil-
ver colloid, it is found that the surface of the PS spheres of both sizes is
surrounded by a large number of silver nanoparticles. The silver nano-
particles clustered on the surface of the PS spheres have a richer SERS
hot spot, which can enhance the Raman signal of the PS sphere.
4. Conclusions
In summary, surface enhanced Raman spectroscopy is used to detect
microplatics and nanoplastics in pure water and seawater. It not only
can effectively detect the plastic particles in the size range from nano-
meter to the micrometer level, but also can identify different types of
plastic particles. For SERS detection of microplastics and nanoplastics
in seawater, it is hardly affected by the complex environment of the
ocean. The SERS-based detection method overcomes the limitations of
microplastics and nanoplastics in liquids and enables detection of
 L. Lv et al. / Science of the Total Environment 728 (2020) 138449
100 nm plastics as low as 40 μg/mL. It provided more possibility for
rapid detection of microplastics and nanoplastics in aquatic environ-
ments s in the future.
CRediT authorship contribution statement
Lulu Lv:Methodology, Investigation, Formal analysis, Writing - orig-
inal draft.Lei He:Software, Validation.Shiqi Jiang:Investigation, Data
curation.Jinjun Chen:Conceptualization, Resources.Chunxia Zhou:
Methodology, Resources.Junhao Qu:Investigation, Resources.Yuqin
Lu:Investigation, Resources.Pengzhi Hong:Supervision, Resources.
Shengli Sun:Conceptualization, Writing - review & editing, Project ad-
ministration.Chengyong Li:Conceptualization, Methodology, Writing -
review & editing, Supervision.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgments
This work is supported by Guangdong Yangfan Program
(201635018), Guangdong Special Support Program (2017TQ04N706),
Science and Technology Planning Project of Zhanjiang City
(2018A02014), Guangdong MEPP Fund (No. GDOE2019A52, R&D on
Degradation of DL-PCBs and Microplastics Exposure to Microbes in
South China Sea (Zhanjiang)), Industrial Development Special Funds
of Dapeng New Area (KY20180202), Southern Marine Science and Engi-
neering Guangdong Laboratory (Zhanjiang) (ZJW-2019-07, ZJW-2019-
08), Innovation and Development Project about Marine Economy Dem-
onstration of Zhanjiang City (2017C8B1), and Training Programs of In-
novation and Entrepreneurship for Undergraduates (CXXL2018008,
CXXL2019302).
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.scitotenv.2020.138449.
References
Araujo, C.F., Nolasco, M.M., Ribeiro, A.M.P., Ribeiro-Claro, P.J.A., 2018. Identification of
microplastics using Raman spectroscopy: latest developments and future prospects.
Water Res. 142, 426–440. https://doi.org/10.1016/j.watres.2018.05.060.
Barnes, D.K., Galgani, F., Thompson, R.C., Barlaz, M., 2009. Accumulation and fragmenta-
tion of plastic debris in global environments. Phil. Trans. R. Soc. B Biol. Sci. 364
(1526), 1985–1998. https://doi.org/10.1098/rstb.2008.0205.
Browne, M.A., Crump, P., Niven, S.J., Teuten, E., Tonkin, A., Galloway, T., Thompson, R.,
2011. Accumulation of microplastic on shorelines woldwide: sources and sinks. Envi-
ron. Sci. Technol. 45 (21), 9175–9179. https://doi.org/10.1021/es201811s.
Cabernard, L., Roscher, L., Lorenz, C., Gerdts, G., Primpke, S., 2018. Comparison of Raman
and Fourier transform infrared spectroscopy for the quantification of microplastics
in the aquatic environment. Environ. Sci. Technol. 52, 13279–13288. https://doi.org/
10.1021/acs.est.8b03438.
Chen, L.-M., Liu, Y.-N., 2012. Ag-nanoparticle-modified single Ag nanowire for detection
of melamine by surface-enhanced Raman spectroscopy. J. Raman Spectrosc. 43,
986–991. https://doi.org/10.1002/jrs.3137.
Cole, M., Galloway, T.S., 2015. Ingestion of nanoplastics and microplastics by pacific oyster
larvae. Environ. Sci. Technol. 49, 14625–14632. https://doi.org/10.1021/acs.
est.5b04099.
da Costa, J.P., Santos, P.S.M., Duarte, A.C., Rocha-Santos, T., 2016. (Nano)plastics in the en-
vironment - sources, fates and effects. Sci. Total Environ. 566, 15–26. https://doi.org/
10.1016/j.scitotenv.2016.05.041.
Dekiff, J.H., Remy, D., Klasmeier, J., Fries, E., 2014. Occurrence and spatial distribution of
microplastics in sediments from Norderney. Environ. Pollut. 186, 248–256. https://
doi.org/10.1016/j.envpol.2013.11.019.
Duemichen, E., Eisentraut, P., Bannick, C.G., Barthel, A.-K., Senz, R., Braun, U., 2017. Fast
identification of microplastics in complex environmental samples by a thermal deg-
radation method. Chemosphere 174, 572–584. https://doi.org/10.1016/j.
chemosphere.2017.02.010.
7
Eerkes-Medrano, D., Thompson, R.C., Aldridge, D.C., 2015. Microplastics in freshwater sys-
tems: a review of the emerging threats, identification of knowledge gaps and
prioritisation of research needs. Water Res. 75, 63–82. https://doi.org/10.1016/j.
watres.2015.02.012.
Eo, S., Hong, S.H., Song, Y.K., Lee, J., Lee, J., Shim, W.J., 2018. Abundance, composition, and
distribution of microplastics larger than 20 μm in sand beaches of South Korea. Envi-
ron. Pollut. 238, 894–902. https://doi.org/10.1016/j.envpol.2018.03.096.
Eriksen, M., Mason, S., Wilson, S., Box, C., Zellers, A., Edwards, W., Farley, H., Amato, S.,
2013. Microplastic pollution in the surface waters of the Laurentian Great Lakes.
Mar. Pollut. Bull. 77, 177–182. https://doi.org/10.1016/j.marpolbul.2013.10.007.
Eriksen, M., Lebreton, L.C.M., Carson, H.S., Thiel, M., Moore, C.J., Borerro, J.C., Galgani, F.,
Ryan, P.G., Reisser, J., 2014. Plastic pollution in the world’s oceans: more than 5 tril-
lion plastic pieces weighing over 250,000 tons afloat at sea. PLoS One 9. https://doi.
org/10.1371/journal.pone.0111913.
Erni-Cassola, G., Gibson, M.I., Thompson, R.C., Christie-Oleza, J.A., 2017. Lost, but found
with nile red: a novel method for detecting and quantifying small microplastics
(1 mm to 20 μm) in environmental samples. Environ. Sci. Technol. 51,
13641–13648. https://doi.org/10.1021/acs.est.7b04512.
Farrell, P., Nelson, K., 2013. Trophic level transfer of microplastic: Mytilus edulis (L.) to
Carcinus maenas (L.). Environ. Pollut. 177, 1–3. https://doi.org/10.1016/j.
envpol.2013.01.046.
Fendall, L.S., Sewell, M.A., 2009. Contributing to marine pollution by washing your face:
microplastics in facial cleansers. Mar. Pollut. Bull. 58, 1225–1228. https://doi.org/
10.1016/j.marpolbul.2009.04.025.
Feng, J., Xu, L., Cui, G., Wu, X., Ma, W., Kuang, H., Xu, C., 2016. Building SERS-active
heteroassemblies for ultrasensitive bisphenol a detection. Biosens. Bioelectron. 81,
138–142. https://doi.org/10.1016/j.bios.2016.02.055.
Feng, Y., Mo, R., Wang, L., Zhou, C., Hong, P., Li, C., 2019. Surface enhanced Raman spec-
troscopy detection of sodium thiocyanate in milk based on the aggregation of Ag
nanoparticles. Sensors 19. https://doi.org/10.3390/s19061363.
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. Environmental Science-Processes & Impacts 15,
1949–1956. https://doi.org/10.1039/c3em00214d.
Gewert, B., Plassmann, M.M., MacLeod, M., 2015. Pathways for degradation of plastic
polymers floating in the marine environment. Environmental Science-Processes &
Impacts 17, 1513–1521. https://doi.org/10.1039/C5EM00207A.
Geyer, R., Jambeck, J.R., Law, K.L., 2017. Production, use, and fate of all plastics ever made.
Sci. Adv. 3. https://doi.org/10.1126/sciadv.1700782.
Gigault, J., Pedrono, B., Maxit, B., Ter Halle, A., 2016. Marine plastic litter: the unana-
lyzed nano-fraction. Environ. Sci. Nano 3, 346–350. https://doi.org/10.1039/
C6EN00008H.
Gigault, J., Halle, A.T., Baudrimont, M., Pascal, P.-Y., Gauffre, F., Phi, T.-L., El Hadri, H., Grassl,
B., Reynaud, S., 2018. Current opinion: what is a nanoplastic? Environmental pollu-
tion (Barking, Essex : 1987) 235, 1030–1034. https://doi.org/10.1016/j.
envpol.2018.01.024.
Harshvardhan, K., Jha, B., 2013. Biodegradation of low-density polyethylene by marine
bacteria from pelagic waters, Arabian Sea, India. Mar. Pollut. Bull. 77, 100–106.
https://doi.org/10.1016/j.marpolbul.2013.10.025.
He, L., Haynes, C., Diez-Gonzalez, F., Labuza, T., 2011. Rapid detection of a foreign protein
in milk using IMS–SERS. J. Raman Spectrosc. 42, 1428–1434. https://doi.org/10.1002/
jrs.2880.
Hernandez, L.M., Yousefi, N., Tufenkji, N., 2017. Are there nanoplastics in your personal
care products? Environ. Sci. Technol. Lett. 4, 280–285. https://doi.org/10.1021/acs.
estlett.7b00187.
Huppertsberg, S., Knepper, T., 2018. Instrumental analysis of microplastics-benefits and
challenges. Anal. Bioanal. Chem. 410. https://doi.org/10.1007/s00216-018-1210-8.
Kaeppler, A., Fischer, M., Scholz-Boettcher, B.M., Oberbeckmann, S., Labrenz, M., Fischer,
D., Eichhorn, K.-J., Voit, B., 2018. Comparison of mu-ATR-FTIR spectroscopy and py-
GCMS as identification tools for microplastic particles and fibers isolated from river
sediments. Anal. Bioanal. Chem. 410, 5313–5327. https://doi.org/10.1007/s00216-
018-1185-5.
Kim, A., Barcelo, S., Williams, S., Li, Z., 2012. Melamine sensing in milk products by using
surface enhanced Raman scattering. Anal. Chem. 84. https://doi.org/10.1021/
ac302025q.
Klaine, S., Koelmans, A., Horne, N., Carley, S., Handy, R., Kapustka, L., Nowack, B., von der
Kammer, F., 2012. Paradigms to assess the environmental impact of manufactured
nanomaterials. Environ. Toxicol. Chem. 31, 3–14. https://doi.org/10.1002/etc.733.
Klaine, S.J., Alvarez, P.J.J., Batley, G.E., Fernandes, T.F., Handy, R.D., Lyon, D.Y., Mahendra, S.,
McLaughlin, M.J., Lead, J.R., 2008. Nanomaterials in the environment: behavior, fate,
bioavailability, and effects. Environ. Toxicol. Chem. 27, 1825–1851. https://doi.org/
10.1897/08-090.1.
Koelmans, A., Besseling, E., Shim, W.J., 2015a. Nanoplastics in the aquatic environment.
Mar. Anthropogenic Litter, 329–344 https://doi.org/10.1007/978-3-319-16510-3_12.
Koelmans, A.A., Besseling, E., Shim, W.J., 2015b. Nanoplastics in the aquatic environment.
Crit. Rev., 325–340 https://doi.org/10.1007/978-3-319-16510-3_12.
Lambert, S., Wagner, M., 2016. Characterisation of nanoplastics during the degradation of
polystyrene. Chemosphere 145, 265–268. https://doi.org/10.1016/j.
chemosphere.2015.11.078.
Law, K.L., Moret-Ferguson, S., Maximenko, N.A., Proskurowski, G., Peacock, E.E., Hafner, J.,
Reddy, C.M., 2010. Plastic accumulation in the North Atlantic subtropical gyre. Sci-
ence 329, 1185–1188. https://doi.org/10.1126/science.1192321.
Law, K.L., Moret-Ferguson, S.E., Goodwin, D.S., Zettler, E.R., De Force, E., Kukulka, T.,
Proskurowski, G., 2014. Distribution of surface plastic debris in the eastern Pacific
Ocean from an 11-year data set. Environ. Sci. Technol. 48, 4732–4738. https://doi.
org/10.1021/es4053076.
8 L. Lv et al. / Science of the Total Environment 728 (2020) 138449
Lee, P.C., Meisel, D., 1982. Adsorption and surface-enhanced Raman of dyes on silver and
god sols. J. Phys. Chem. 86. https://doi.org/10.1016/j.colsurfa.2011.12.048.
Lenz, R., Enders, K., Stedmon, C.A., Mackenzie, D.M.A., Nielsen, T.G., 2015. A critical assess-
ment of visual identification of marine microplastic using Raman spectroscopy for
analysis improvement. Mar. Pollut. Bull. 100, 82–91. https://doi.org/10.1016/j.
marpolbul.2015.09.026.
Li, A., Tang, L., Song, D., Song, S., Ma, W., Xu, L., Kuang, H., Wu, X., Liu, L., Chen, X., Xu, C.,
2016. A SERS-active sensor based on heterogeneous gold nanostar core-silver nano-
particle satellite assemblies for ultrasensitive detection of aflatoxinB1. Nanoscale 8,
1873–1878. https://doi.org/10.1039/C5NR08372A.
Lin, X., Hasi, W.-L.-J., Lou, X.-T., Lin, S., Yang, F., Jia, B.-S., Cui, Y., Ba, D.-X., Lin, D.-Y., Lu, Z.-
W., 2014. Rapid and simple detection of sodium thiocyanate in milk using surface-
enhanced Raman spectroscopy based on silver aggregates. J. Raman Spectrosc. 45,
162–167. https://doi.org/10.1002/jrs.4436.
Lodge, T.P., 2017. Celebrating 50 years of macromolecules. Macromolecules 50 (24),
9525–9527. https://doi.org/10.1021/acs.macromol.7b02507.
Ma, W., Sun, M., Xu, L., Wang, L., Kuang, H., Xu, C., 2013. A SERS active gold nanostar dimer
for mercury ion detection. Chem. Commun. 49, 4989–4991. https://doi.org/10.1039/
C3CC39087J.
Mani, T., Hauk, A., Walter, U., Burkhardt-Holm, P., 2015. Microplastics profile along the
Rhine River. Sci. Rep. 5. https://doi.org/10.1038/srep17988.
Mecker-Pogue, L., Tyner, K., Kauffman, J., Arzhantsev, S., Mans, D., Gryniewicz, C., 2012.
Selective melamine detection in multiple sample matrices with a portable Raman in-
strument using surface enhanced Raman spectroscopy-active gold nanoparticles.
Anal. Chim. Acta 733, 48–55. https://doi.org/10.1016/j.aca.2012.05.001.
Mintenig, S.M., Bäuerlein, P.S., Koelmans, A.A., Dekker, S.C., van Wezel, A.P., 2018. Closing
the gap between small and smaller: towards a framework to analyse nano- and
microplastics in aqueous environmental samples. Environmental Science: Nano 5,
1640–1649. https://doi.org/10.1039/c8en00186c.
Nam Ngoc, P., Zalouk-Vergnoux, A., Kamari, A., Mouneyrac, C., Amiard, F., Poirier, L.,
Lagarde, F., 2018. Quantification and characterization of microplastics in blue mussels
(Mytilus edulis): protocol setup and preliminary data on the contamination of the
French Atlantic coast. Environ. Sci. Pollut. Res. 25, 6135–6144. https://doi.org/
10.1007/s11356-017-8862-3.
Nandiyanto, A., Suhendi, A., Ogi, T., Iwaki, T., Okuyama, K., 2012. Synthesis of additive-free
cationic polystyrene particles with controllable size for hollow template applications.
Colloids Surf. A Physicochem. Eng. Asp. 396, 96–105. https://doi.org/10.1016/j.
colsurfa.2011.12.048.
Ngoc Nam, P., Zalouk-Vergnoux, A., Kamari, A., Mouneyrac, C., Amiard, F., Poirier, L.,
Lagarde, F., 2017. Quantification and characterization of microplastics in blue mussels
(Mytilus edulis): protocol setup and preliminary data on the contamination of the
French Atlantic coast. Environ. Sci. Pollut. Res. Int. 25. https://doi.org/10.1007/
s11356-017-8862-3.
Peiponen, K.-E., Raty, J., Ishaq, U., Pelisset, S., Ali, R., 2019. Outlook on optical identification
of micro- and nanoplastics in aquatic environments. Chemosphere 214, 424–429.
https://doi.org/10.1016/j.chemosphere.2018.09.111.
Revel, M., Châtel, A., Mouneyrac, C., 2018. Micro(nano)plastics: a threat to human health?
Current Opinion in Environmental Science & Health 1, 17–23. https://doi.org/
10.1016/j.coesh.2017.10.003.
Schirinzi, G.F., Llorca, M., Sero, R., Moyano, E., Barcelo, D., Abad, E., Farre, M., 2019. Trace
analysis of polystyrene microplastics in natural waters. Chemosphere 236, 124321.
https://doi.org/10.1016/j.chemosphere.2019.07.052.
Schlining, K., von Thun, S., Kuhnz, L., Schlining, B., Lundsten, L., Jacobsen Stout, N., Chaney,
L., Connor, J., 2013. Debris in the deep: using a 22-year video annotation database to
survey marine litter in Monterey canyon, Central California, USA. Deep-Sea Res. I
Oceanogr. Res. Pap. 79, 96–105. https://doi.org/10.1016/j.dsr.2013.05.006.
Setala, O., Fleming-Lehtinen, V., Lehtiniemi, M., 2014. Ingestion and transfer of
microplastics in the planktonic food web. Environ. Pollut. 185, 77–83. https://doi.
org/10.1016/j.envpol.2013.10.013.
Shim, W.J., Hong, S.H., Eo, S.E., 2017. Identification methods in microplastic analysis: a re-
view. Anal. Methods 9, 1384–1391. https://doi.org/10.1039/C6AY02558G.
Suaria, G., Avio, C.G., Mineo, A., Lattin, G., Magaldi, M., Belmonte, G., Moore, C., Regoli, F.,
Aliani, S., 2016. The Mediterranean plastic soup: synthetic polymers in Mediterra-
nean surface waters. Sci. Rep. 6, 37551. https://doi.org/10.1038/srep37551.
Ter Halle, A., Jeanneau, L., Martignac, M., Jarde, E., Pedrono, B., Brach, L., Gigault, J., 2017.
Nanoplastic in the North Atlantic subtropical gyre. Environ. Sci. Technol. 51,
13689–13697. https://doi.org/10.1021/acs.est.7b03667.
Vo-Dinh, T., Allain, L., Stokes, D., 2002. Cancer gene detection using surface-enhanced
Raman scattering (SERS). J. Raman Spectrosc. 33, 511–516. https://doi.org/10.1002/
jrs.883.
Wang, W., Wang, J., 2018. Investigation of microplastics in aquatic environments: an
overview of the methods used, from field sampling to laboratory analysis. Trac-
Trends in Analytical Chemistry 108, 195–202. https://doi.org/10.1016/j.
trac.2018.08.026.
Wright, S.L., Kelly, F.J., 2017. Plastic and human health: a micro issue? Environ. Sci.
Technol. 51, 6634–6647. https://doi.org/10.1021/acs.est.7b00423.
Xie, W., Schlücker, S., 2013. Medical applications of surface-enhanced Raman scattering.
Physical Chem. Chem. Phys. 15. https://doi.org/10.1039/C3CP43858A.
Xu, L., Yin, H., Ma, W., Kuang, H., Wang, L., Xu, C., 2015. Ultrasensitive SERS detection of
mercury based on the assembled gold nanochains. Biosens. Bioelectron. 67,
472–476. https://doi.org/10.1016/j.bios.2014.08.088.