2020 - Marti - The Colours of The Ocean Plastics

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Contaminants in Aquatic and Terrestrial Environments

The colours of the ocean plastics
Elisa Marti, Cecilia Martin, Matteo Galli, Fidel Echevarría, Carlos M. Duarte, and A. Cozar
Environ. Sci. Technol., Just Accepted Manuscript • DOI: 10.1021/acs.est.9b06400 • Publication Date (Web): 11 May 2020
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Page 1 of 25 Environmental Science & Technology
The colours of the ocean plastics
Elisa Martí1*, Cecilia Martin2, Matteo Galli3, Fidel Echevarría1, Carlos M. Duarte2,
Andrés Cózar1*
1 Departamento de Biología, Campus de Excelencia Internacional del Mar (CEIMAR),
Instituto Universitario de Investigaciones Marinas (INMAR), Universidad de Cádiz, E-
11510 Puerto Real, Spain
2 King Abdullah University of Science and Technology, Red Sea Research Center,
Thuwal 23955-6900, Kingdom of Saudi Arabia
3 Department of Physical Sciences, Earth and Environment, University of Siena, Via
P.A. Mattioli 4, 53100, Siena, Italy
* Corresponding authors: elisa.marti@uca.es; andres.cozar@uca.es
1
ACS Paragon Plus Environment
Environmental Science & Technology Page 2 of 25
1 Abstract
2 Characterisation of the colour is often included in studies on plastic pollution. However,
3 the comparability and relevance of this information is limited by methodology or
4 observer subjectivity. Based on the analysis of thousands of floating plastic fragments
5 from a global collection, here we propose a systematic semi-automatic method to
6 analyse colours by using a reference palette of 120 Pantone colours. The most abundant
7 colours were white and transparent/translucent (47 %), yellow and brown (26 %) and
8 blue-like (9 %). The white colour increased in the smallest pieces (< 5 mm) and far
9 from coastal sources (> 500 km). Both fragmentation and discolouration of ocean
10 plastics may occur because of longer exposure time to sunlight in nature. In addition,
11 yellow items peaked at around 1 cm and brown colours at around 1 mm, supporting the
12 notion that yellowing precedes tanning in the aging process, which is paralleled by
13 fragmentation. Apart from the effects of the weathering, our results suggest a second-
14 order modulation of the colour distributions of marine plastic microplastics by the
15 selective action of visual predators. The present work provides methodological tools
16 and a wide empirical background to further the interpretation and applicability of the
17 colour information on ocean plastics.
18
19 Table of Contents (TOC) Art
2
20 Introduction
21 Plastic pollution spreads throughout the global ocean (Cózar et al. 2014; Eriksen et al.
22 2014; van Sebille et al. 2015). The weathering of plastic debris in the environment leads
23 to a progressive degradation of the chemical bond structure of the polymers (Brandon et
24 al. 2016; ter Halle et al. 2017). Despite this degradation, plastics hardly disappear in
25 nature. Plastic objects break down into smaller and smaller pieces, which are
26 transported by currents across the oceans to reach even the most remote areas (Zhang
27 2017; Cózar et al. 2017).
28 We have a limited knowledge about the transformation of the plastic debris in the
29 environment. The stages of weathering or the time scales of degradation, fragmentation
30 and transport in the marine environment are poorly understood. While staying in inland
31 environments, stranding on the shores or floating at the ocean surface, plastic debris
32 undergoes photo-oxidation and changes in the absorbance and reflection of light,
33 acquiring scratched surface and shifts in its tonality (Weinstein et al. 2016; ter Halle et
34 al. 2016; Brandon et al. 2016). Due to weathering, discolouration (Pospísil et al. 2002)
35 and the development of yellowish colours are common features on plastics aged in the
36 environment (Andrady et al. 1992; Singh et al. 2001; Yousif & Haddad 2013; Pastorelli
37 et al. 2014). Thus, the size as well as the colour of oceanic plastic debris may reflect
38 their degradation stages and be potentially used as proxies of exposure time in the
39 environment.
40 Numerous studies have accounted for the plastic colour, using from simple sorting with
41 4 colour categories (Shaw & Day 1994; Barrows et al. 2017) to more detailed
42 classifications based on up to 32 colours (Blair Crawford & Quinn 2017). There is a
43 pressing demand for replicable and objective methods to analyse the colour of ocean
3
44 plastic (Hartmann et al. 2019). However, standard procedures for colour categorisation
45 are lacking thus far. At present, the process of categorisation of colours is highly
46 subjective and depends, among others, on visual capabilities and experience of
47 observer. The lack of standardized methods imposes significant difficulties to compare
48 studies and achieve robust conclusions.
49 In the present work, we characterized the colour of thousands of items collected with
50 surface trawling plankton nets at the global scale. Our main goal is to use this extensive
51 analysis to define a consistent methodological tool to characterize colour in marine
52 plastic samples. Additionally, we provide a robust benchmark against which future data
53 sets can be compared to and explore potential insights the colour information may offer
54 on sources and fate of marine plastics. Based on weathering experiments (Andrady
55 2017), we expect photo-oxidative damage to lead to increasing fractions of white and
56 yellow-brown colours in ocean plastics with increasing exposure time in the
57 environment, parallel to reduction in their sizes (ter Halle et al. 2017). Likewise, the
58 transport of plastic fragments to distant open-ocean accumulations is proposed to be a
59 time-dependent process, with the fragments far from shore being, on average, older than
60 nearby ones (Brandon et al. 2016). Here we explore the relationships between colours of
61 marine plastic debris with particle size and distance to the coast. These relationships
62 may, in turn, support the use of colour information to provide insights onto the nature
63 and dynamics of marine plastic litter.
64 Material and Methods
65 Marine plastic items were collected by surface trawling nets in the circumnavigation
66 cruise “Malaspina 2010” together with other regional surveys carried out from 2010 to
67 2017 (Table 1). Samplings covered open-ocean plastic accumulation zones located in
4
68 each of the five sub-tropical gyres (North Pacific, North Atlantic, South Pacific, South
69 Atlantic and Indian Ocean) as well as semi-enclosed regions (Mediterranean, Arctic
70 Ocean, Red Sea and Bay of Biscay). Overall, 8,849 items with sizes from 0.2 mm to 15
71 cm were collected from surface waters with distance to land ranging from 0.2 to 3,000
72 km (Fig. S1). Distances were measured to the nearest continental coasts, excluding the
73 coasts of small oceanic islands. Regarding the polymeric composition of the plastic
74 items, a previous work carried out on a subset of 694 items showed that 94.2 % of the
75 fragments were made of polyethylene (PE) and polypropylene (PP) (Serranti et al.
76 2018). Other major commodity plastics such as polystyrene (PS), polyethylene
77 terephthalate (PET) or polyvinyl chloride (PVC) were barely represented in the dataset.
78
79 Table 1. Summary of sampling regions, date and number of plastic items (N =
80 8,849) collected using surface trawling nets. Location of sampling sites is shown
81 in Figure S1.
Region/Cruise Date No. Items Ref.

Malaspina
Circumnavigation and 2010 - 2011 2994 Cózar et al. 2014
others
Mediterranean Sea 2013 3369 Cózar et al. 2015
Arctic Ocean 2013 775 Cózar et al. 2017
Red Sea 2016 - 2017 297 Martí et al. 2017
Atlantic Ocean 2015 452 This study
Strait of Gibraltar 2015 525 This study
Bay of Biscay 2017 437 This study
82
83 Colour of all plastic items was categorized using a reference palette with 13 main
84 colours and 9 hues from black to white, besides the transparent/translucent, accounting
85 for a total of 120 possible colour codes (120-Palette, Fig. 1; see Table S1 for RGB
5
86 scores and Pantone´s codes). Based on the experience acquired during the analyses, we
87 selected a 120-colours scheme in order to find a balanced equilibrium between
88 analytical effort and level of detail provided by the results. Human capacity to assign
89 colours may vary between observers since it depends on the particular features of the
90 observer´s retina and prior experience. By using a reference palette, here we aim to
91 minimize differences in the observer´s capacity to assign colours. Two alternatives are
92 proposed to the palette-assisted assignment of colours, a visual method and a digital
93 semi-automatic method, both providing comparable results (see Supporting Methods).
94 Both methods are applied once the plastics were washed and dried. The reflectance of
95 wet and dry plastic is different, leading to more stable data from dry samples. Therefore,
96 all plastic pieces were washed with ultrapure water and analysed once they were
97 completely dry in order to render the results comparable and the analyses reproducible.
6
98
99 Figure 1. Palette of 120 colour codes (120-Palette) that includes 13 colours (vertical
100 axis) and 9 hues (horizontal axis) from black to white, besides the
101 transparent/translucent. RGB scores and Pantone´s codes are shown in Table S1. A
102 simplified version of this reference palette can be derived from the aggregation of dark,
103 medium and light hues (42-Palette, see Methods).
104
105 Visual palette-assisted method for colour assignment
106 Dried plastic items were organized into glass Petri dishes by main colours. The printed
107 colour palette was placed under the Petri dish as reference and colour codes were
108 assigned to the items by visual similarity (Fig. S2). To ensure the replicability of the
7
109 printed version of the 120-Palette, we also provide Pantone´s codes for all colours in the
110 reference palette (Table S1). When the colour of an item was markedly heterogeneous,
111 for instance due to biofouling, an additional secondary colour was registered. A low
112 percentage of double colour assignment was needed (< 2 % of the total), and finally the
113 analyses were exclusively based on the primary colour.
114 Digital semi-automatic method for colour assignment
115 From the experience acquired here, we developed a digital method for colour
116 assignment to facilitate future analyses. A subsample of plastic items (N = 103) was
117 processed by the digital method for comparative purposes. Dry plastic items were
118 photographed with a high-resolution camera (NIKON D810, exposure 1/15 sec,
119 aperture f/4.5 and no flash) and examined using ImageJ (https://imagej.nih.gov/ij/). This
120 software, like other common image analysers, provides a digital colour categorization
121 through a combination of scores for red, green and blue colours (RGB) on a 0 to 255
122 scale. Thus, RGB palette comprises 16,777,216 colours (2563). To convert any RGB
123 score into one of the 120 colours in the reference palette, RGB scores for each of the
124 colour codes from the 120-Palette were used as reference nodes in the three-
125 dimensional RGB colour space (Table S1; Fig. S3). Therefore, by identifying the
126 reference node at the minimum distance from the RGB scores of a given item, one of
127 the 120 colours of the reference palette may be automatically assigned to any RGB
128 score by proximity in the colour space (see Supporting Methods).
129 Size and shape of the plastic items
130 The size of the plastics was measured from the high-resolution photographs using
131 ImageJ software. Items were also classified according to shape/origin in six types: raw
132 industrial pellets, microbeads (likely derived from cosmetic and cleansing products),
8
133 films (mostly derived from discarded bags and wrappings), foamed plastic, rigid (thick-
134 walled) fragments, and fishing lines (likely also derived from nets). The colour pattern
135 for each plastic type was examined using principal components analysis (PCA) from the
136 SPSS statistical package (version 24). Due to the low representation of some colours,
137 we used a simplified version of the reference palette for the PCA (36-Palette). This
138 version grouped in dark (hues 1, 2 y 3, Fig. 1), medium (hues 4, 5 y 6) and light (hues 7,
139 8 and 9), and grouped violet, magenta and pink (which represented only 0.88 % in our
140 dataset) as wide magenta.
141 Possible trends in the colour spectra were examined in relation to plastic size and
142 distance to the coast, explored here as potential proxies of the age of plastic litter in the
143 marine environment (Brandon et al. 2016; ter Halle et al. 2017). A Single Value
144 Decomposition (SVD) was used to infer which colours showed similar covariance with
145 size or distance to land. SVD provides a series of eigenfunctions associated with
146 dominant modes of colour variability. For each eigenfunction, the related eigenvalue
147 indicates the fraction of the total variability explained. Significant eigenfunctions were
148 then compared to the variability patterns of the percentage of each colour in relation to
149 size or distance to nearest continental coast. Using the Pearson correlation coefficient,
150 the colour codes were linked to the main eigenfunctions and gathered into groups
151 having similar co-variation. Colours that were not significantly related to any of the
152 main eigenfunctions were represented jointly as “Rest of colours”.
153 Results
154 As a first step, the colour spectrum of the complete collection of plastic items (n =
155 8,849) was analysed by using the visual method assisted with the reference palette of
156 120 colour codes (120-Palette). We found a wide range of colours (103 of the 120 used
9
157 in the palette), with a prevalence of white, transparent/translucent, black, blue-like and
158 yellow-brown colours. Rigid (thick-walled) fragments were the most abundant plastic
159 type, accounting for 84 % of the total items analysed (Table S2). Rigid fragments also
160 showed the widest colour diversity. Film-type plastics were mainly related to
161 transparent/translucent items, while fishing lines were highly related to blue, turquoise
162 and green colours (Fig. 2, Fig. S6).
163
164 Figure 2. Biplot for the PCA analysis of plastic type and colour. All items (N = 8,849)
165 are plotted in relation to the first two principal components, accounting for 83.2 % of
166 the variability (χ2 = 1,892, df = 175, p < 0.05). Fishing lines were located in the upper
167 left, related to BD, TL, TM and ED colours (see left legend). Films were located in the
168 upper right, being mainly transparent/translucent (TRANS). Data used for this figure are
169 provided in Table S2 and graphed in bar charts in Fig. S6.
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170
171 Our analyses focused on the colour-hue matrix for rigid fragments since this plastic type
172 was by far the most abundant in the samples. Apart from white, transparent/translucent
173 and black, which were particularly common (31 %, 16 % and 7 % of the total,
174 respectively), there were three zones in the matrix showing high abundance of items
175 (Fig. 3). Amber colours, that is, those in the range of yellow-brown-orange, added a 28
176 % of the total items, bluish-green colours encompassed an 11 %, and grey colours
177 accounted for a 5 %. The lowest proportion was detected in reddish tonalities (violet,
178 magenta, pink and red), which gathered just a 2 % of the total of rigid fragments.
179 Projecting data on the hue axis of the matrix (Fig. 3), we found maxima at yellow and
180 brown colours and an increasing trend in abundance towards the lightest hues.
181
182
183
184 Figure 3. Proportions of colour for rigid plastic fragments (n = 7,395) in the 120-
185 Palette. Percentages of hues and colours are added and projected on left and back
186 planes, respectively, showing increasing percentages towards the lightest hues and
187 maxima at yellow and brown colours. Note that the graph excludes white,
188 transparent/translucent and black (31 %, 16 % and 7 %, respectively). Colour
11
189 assignment was carried out by the visual palette-assisted method. Numerical
190 percentages for each colour are provided in Table S3.
191
192 The covariance of colours in relation to distance to land and plastic size was determined
193 through SVDs using only rigid fragments for the analysis (Fig. 4). In terms of distance
194 to land, the four main principal components explained 93 % of the colour variability.
195 PC1 was related to the variability of white and light-cyan colours. The abundance of
196 these colours was around 15 % near the shores and rapidly increased from 500 km to
197 coast, accounting for 70 % of all items at distances larger than 2,000 km (Fig. 4A). The
198 offshore increase in the proportion of white and light-cyan was not observable just from
199 the semienclosed-seas dataset (Mediterranean and Red Sea, Fig. S7), likely because all
200 these sampling sites were located relatively close to land (< 500 km). Likewise, the
201 group of “Rest of colours” was mostly represented by medium colour tonality in the
202 semi-enclosed seas (53 %) while light colour tonality was predominant into this group
203 in the open-ocean dataset (55 %). “Rest of colours” showed an increasing trend
204 landward, contrary to the white colour.
205 Four components explained 82 % of the variability of colours in relation to the plastic
206 size gradient (Fig. 4B). White colour predominated throughout the whole size gradient,
207 especially in the open-ocean dataset (Fig S8), which accounted for more distant zones
208 from land. An increasing trend in the proportion of white items towards small sizes was
209 found in the joint dataset, particularly from 0.5 to 5 millimeter in size. Interestingly, the
210 proportion of yellow colours showed maxima around 10 mm in size, while brown
211 colours peaked around 1 mm. Moreover, yellow and brown colours showed similar
12
212 patterns in both datasets, open ocean and semi-enclosed seas, in spite of the differences
213 in the load of white fragments.
214
215 Figure 4. Percentages of colour in relation to distance to land (A) and plastic size (B).
216 Percentages were calculated for moving averages of 200 rigid plastic fragments. Single
217 Value Decomposition (SVD) and Pearson correlation coefficients were used to group
218 colours showing similar covariance in relation to distance to land and size. Figure
219 legends show the main colour groups. “Rest of colours” was dominated by bluish
220 colours (cyan, sky and blue, > 22 %) in B, while it comprised a higher diversity of
221 colours in A. Colour assignment was carried out by the visual palette-assisted method.
222
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223 Discussion
224 The present study provides the most exhaustive and comprehensive analysis of the
225 colour of marine plastic debris. A total of 8,849 floating items collected from around the
226 world were classified by using a 120-colour palette, accounting for differences in plastic
227 typology, size and distance to land. Rigid fragment was the most abundant plastic type
228 (84 % of the total items), in agreement with observations of other authors in oceanic
229 surface waters (Morét-Ferguson et al. 2010; Reisser et al. 2015). Therefore, the analyses
230 of colour patterns in relation to particle size and distance to land focused on the
231 collection of rigid fragments in order to avoid confounding effects of changes in plastic
232 types on colour distributions (Fig. 2).
233 Photo-oxidation is a known process that induces changes in both mechanical properties
234 and colour of the plastic polymers. Comparing recently-manufactured objects,
235 mesoplastic pieces (5 - 20 mm) and microplastics (0.3 - 5 mm), ter Halle et al. (2017)
236 found a progressive oxidative degradation of the polymers, with evident changes in
237 properties such as crystallinity and molar mass towards smaller plastic sizes. Oxidative
238 degradation of plastics during weathering determines the ease of crack formation and
239 fragmentation. In parallel, photo-oxidation generally causes changes in plastic colour,
240 with experimental weathering tests leading to the expectation of gradual change towards
241 light colours (discolouration or whitening), while the accumulation of degradation
242 products in the plastic matrix as result of the oxidation typically gives yellow or amber
243 colours (yellowing and tanning, respectively) (Andrady 2017).
244 Open-ocean samples comprised a considerably high fraction of white items (26 %),
245 much higher than those in semi-enclosed seas (6 %). The proportion of white items
246 increased from 15 % to 70 % beyond 500 km from land (Fig. 4A), supporting the
14
247 hypothesis of progressive discolouration as plastic debris moves away offshore. Based
248 on the changes in the chemical bonds of polyethylene particles, Brandon et al. (2016)
249 inferred exposure times longer than 18 months for floating particles collected far from
250 shores, into the accumulation zone of the North Pacific Central Gyre, while particles
251 sampled in both intermediate and near-shore waters could have generally weathered less
252 than 18 months. The relationship between white items and particle size showed the
253 highest proportions of white particles in the microplastic size range, from 0.5 to 5 mm
254 (Fig. 4B). This pattern was particularly evident in the open-ocean dataset (Fig. S8),
255 likely because the semi-enclosed dataset was biased towards samples collected at
256 relatively short distances from the coast (< 500 km to land) and its load of white items
257 was relatively low.
258 Like white items, the fraction of light-cyan items also increased far from shore (Fig.
259 4A). Light-cyan colour may be derived from the discolouration of bluish colours.
260 Indeed, we found a considerable number of bluish items (cyan, sky and blue),
261 contrasting with the very low number of reddish items (violet, magenta, pink and red)
262 (Fig. 3). Interestingly, Shaw and Day (1994) found similar colour-dependent loss of
263 floating microplastics in the North Pacific Ocean. A possible explanation could be
264 related to a lower removal of bluish plastics from the ocean surface by plastic-ingesting
265 predators such as seabirds or vertically migratory fish, as suggested by Shaw and Day
266 (1994). Blue has been suggested as a common camouflage colour in the ocean surface
267 (Hudelson 2011; Umbers 2012). Thus, blue pigmentation is commonly used for the
268 organisms living on the surface, with successful blue-coloured life forms in the neuston
269 such as Velella velella, Porpita porpita, Glaucus atlanticus or many species of oceanic
270 copepods (Fig. S9). A higher probability of detection and ingestion of items with non-
271 blue colours, like red coloured, by visual surface predators would lead to a progressive
15
272 enrichment in blue microplastics on the ocean surface. In lakes without
273 zooplanktivorous pressure, a more cost-effective red pigmentation is used by the
274 zooplankton to protect themselves against UV radiation; however, UV-protective
275 pigment is turned from red to blue in order to reduce the losses by predation when
276 zooplanktivorous fishes increase (Hudelson 2011). Therefore, if the removal of
277 microplastics from the surface by visual predators becomes a significant sink process in
278 the open ocean, we could expect an increase in the proportion of blue-like colours,
279 especially towards the smallest sizes, as observed (Shaw & Day 1994), because the
280 effect on the colour distribution would be cumulative. However, we note that some
281 visual predators have been shown to have preferential ingestion for blue prey, as in the
282 case of small planktivorous fishes along the coast of Easter Island, in the South Pacific
283 Subtropical Gyre. Ory et al. (2017) found that these fishes regularly ingest blue plastic
284 fragments that resemble their common prey items, blue copepods. A second possible
285 explanation for the increase of light cyan items towards remote areas and among the
286 smallest items could be that the fragmentation of fishing lines, abundant in bluish
287 colours (Fig. 2, Fig. S6) is generating small pieces not identifiable as fishing-line
288 fragments, being identified as rigid fragments and therefore increasing the abundance of
289 light-cyan items. Finally, a particular resistance of the bluish colours to the UV
290 radiation and associated nanofragmentation could also explain the observed patterns.
291 However, we know little about the effects of additives on plastic photodecomposition,
292 the colour-selectivity of visual active predators (Ryan 1987), or possible colour shifts of
293 ingested plastic after with gut acid treatment. Consequently, no hypothesis can be ruled
294 out yet in relation to the abundance of bluish plastics at sea.
295 Yellow and brown colours were also particularly abundant in our collection of marine
296 plastic debris (Fig. 3). The phenomena of yellowing and tanning are commonly
16
297 observed in all of the most common plastic polymers, including PE, PP, PS, PET or
298 PVC (Brandon et al. 2016, Andrady 2017). The degree of yellow or brown colour is
299 associated with the amount of products resulting from the photo-oxidation, hence the
300 extent of the weathering (Andrady 1997; ter Halle et al. 2016). The oxidative process
301 may act on the polymer itself (e.g. PVC, PC, PS) as well as on the thermal stabilizers
302 added to the plastic resin (Andrady 2017). The dependence of the yellow-brown
303 colouration on the nature and concentration of additives in the plastic matrix (Cooper &
304 Corcoran 2010; Fisner et al. 2017) makes the analyses based on these colours complex
305 due to the plethora of additives in the market. However, our analysis provides a
306 consistent pattern in the abundance of yellow and brown items across the plastic size
307 gradient (Fig. 4B). Light yellow items reached the highest proportions in plastic items
308 around 1 centimetre in size, while fragments around 1 millimetre tended to be light
309 brown in colour (Fig. 4B). This pattern was consistent across independent datasets,
310 from open-ocean and semi-enclosed seas, which renders this a robust result (Fig. S8).
311 We must note this analysis focused on rigid floating fragments. As in other sea-surface
312 datasets, only two low-density polymers (PP and PE) accounted for most of the
313 fragments in our dataset (Serranti et al. 2018, Pedrotti et al. 2016), which must reduce
314 the variability of yellowing and tanning patterns expected for the diversity of plastic
315 resins in the market.
316 Interestingly, weathering testing with plastic materials suggests a typical progressive
317 shift from white to yellow to brownish colour upon an extended solar exposure
318 (Andrady 2017), matching with our finding of a succession of the peaks of yellow and
319 brown items across the marine plastic size gradient. Combining colour and size
320 distributions of marine plastic items provides a pathway to infer age for microplastic
321 samples by using, for example, ratios of colours or RGB scores (Table S1). Indeed, the
17
322 combination of RGB scores into yellowing indexes is commonly used by the polymer
323 industry to assess the degradation of the plastic materials exposed to solar radiation (e.g.
324 Andrady 2017).
325 The experience acquired with the present work has allowed defining a systematic
326 method to the colour analysis based on a 120-colour palette as reference to assign
327 colours. The design of the reference palette (120-Palette) was optimized to provide an
328 operational method, in terms of observer’s capacity and analytical effort, as well as a
329 thorough colour categorization able to be applied to other studies or purposes, including
330 unexplored sites with potentially different colour spectra or other applications that
331 cannot be anticipated. In this study, we simplified the colour scheme of the 120-Palette
332 for the statistical PCA analysis, due to the low representativeness of some colours in our
333 dataset. The samples might be processed, if needed, using a simplified reference palette
334 derived from the aggregation of dark, medium and light hues (42-Palette, see Methods).
335 However, working with a 120-Palette was not excessively demanding and the gathering
336 of additional information by using the 120-Palette would contribute to expand the
337 standardized high-detail colour dataset provided here (Table S3). The current baseline
338 dataset was fully obtained from the visual method. Based on this experience, we
339 developed a comparable alternative digital method to advance in the automatization of
340 the colour assessment if future analyses (see Supporting Methods).
341 In addition to providing tools and an empirical benchmark for analysing colours, here
342 we discussed about the processes controlling the colour distribution of the marine
343 plastic. Colour changes (whitening, yellowing and tanning) possibly related to the
344 photo-oxidative degradation were identified as main processes modulating the colour
345 spectrum of marine plastic. Our results provide a first evidence for a relationship
346 between distance to land, size and colour of marine plastic debris, suggesting that colour
18
347 may provide a qualitative proxy for the age of marine plastic samples, particularly when
348 combined with size. White, yellow and brown colours, or a combination of them a priori
349 appear as the most suitable colours to devise information about ageing. However,
350 further and more focused research is required to assess whether colour and size
351 distributions can be jointly used to provide a robust proxy of the age of plastics at sea.
352 ASSOCIATED CONTENT
353 Supporting information. Additional figures and tables.
354 AUTHOR INFORMATION
355 * Corresponding authors: elisa.marti@uca.es; andres.cozar@uca.es
356 Author Contributions
357 EM, FE, CD and AC conceived and designed the study; EM, CM and MG processed the
358 samples; EM, FE, CD and AC contributed to analysis and interpretation of data; EM
359 and AC drafted the manuscript and EM, CM, MG, FE, CD and AC reviewed the
360 manuscript.
361 Conflict of Interest Statement
362 The authors declare no competing financial interest.
363 Acknowledgments
364 This study is an outcome of Malaspina 2010 expedition (Consolider-Ingenio 2010,
365 CSD2008-00077). E.M. was supported by the Campus de Excelencia Internacional del
366 Mar (CEIMAR) through a PhD Research Project Grant. We received additional support
367 from PLASTREND (BBVA Foundation) and MIDaS (CTM2016- 77106-R,
19
368 AEI/FEDER/UE) projects. We thank personnel from Malaspina 2010 expedition,
369 MEDSea (EU contract number FP7-2010-265103), TARA Arctic Ocean, MAFIA
370 (Migrants and Active Flux In the Atlantic Ocean), MEGAN (Mesoscale and
371 submesoscale processes in the Strait of Gibraltar: The Trafalgar-Alborán connection),
372 Seagrass and Mangrove Cruise through Red Sea and ETO y NST Cruise along Bay of
373 Biscay, for the help with the sample collection.
374 References
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24
493
494
25
Supporting Information for

“The Colors of the Ocean Plastics”

Elisa Martí1.*. Cecilia Martin2. Matteo Galli3. Fidel Echevarría1. Carlos M. Duarte2.
Andrés Cózar1.*

1
Departamento de Biología. Campus de Excelencia Internacional del Mar (CEIMAR).
Instituto Universitario de Investigaciones Marinas (INMAR). Universidad de
Cádiz. E-11510 Puerto Real. Spain
2
King Abdullah University of Science and Technology. Red Sea Research Center.
Thuwal 23955-6900. Kingdom of Saudi Arabia
3
Department of Physical Sciences. Earth and Environment. University of Siena. Via P.A.
Mattioli 4. 53100. Siena. Italy

*Corresponding authors. Email: elisa.marti@uca.es; andres.cozar@uca.es

This document includes:

Supporting Methods
Figures S1 to S9
Tables S1 to S3

S1

Figure S1. Location of sampling sites on the global ocean surface.

Supporting Methods:
All plastic items were categorized by visual palette-assisted method (Fig. S2).
Nevertheless, a subsample of plastic items (N = 103) was also processed by the digital
semi-automatic method for comparative purposes.

Figure S2: Modus operandi for the visual palette-assisted method for colour assignment. A
printed colour palette (defined by Pantone´s codes in Table S1) is placed under the Petri dish
as reference, and colour codes are assigned to the plastic items by visual similarity.

To assign colours from digital images, RGB scores for the colours in the 120-Palette
were used as reference nodes in the three-dimensional RGB colour space (Fig. S3,
S2
Table S1). Plastic items were photographed with a high-resolution camera (NIKON
D810) and natural light. Images were captured with an exposure time of 1/15 second,
a wide aperture of f/4.5 and no flash. Focal length and ISO were variable. RGB scores
for plastic items were obtained after averaging the colour of each item (Fig. S4). RGB
scores were obtained from digital photographs and image analysis software (Fig. S4).
Images were processed through an image analysis software (e.g. ImageJ).

Figure S3. Location of the reference colours of the 120-Palette in the RGB colour space.

Processing steps in ImageJ software are:
• “makePolygon” or “wand tool” to select plastic item partially or completely.
• Then, “average colour” to average pixels contained in the selection.
• Finally, “dropper tool” to get RGB values.

ImageJ macro for each plastic item:
• //setTool("polygon"); or //setTool("wand");
• //run("Average Color");
• //setTool("dropper");
S3

Figure S4: Example of the colour averaging process with from original image (A) to averaged
RGB (B and C). Note that the method for semi-automatic assignment of colour codes requires
the previous removal of transparent items and particular treatment for items with very
heterogeneous colour (e.g. biofouled items). Here, we show examples for these cases: the
transparent item (middle) is classified manually and the item showing heterogeneous
colouration (right) is processed by adding an additional secondary colour (C) (see Methods).
Likewise, items including biofouling could show different colours. Scale bar indicates 5 mm.

Once RGB scores for 120 reference nodes (Rn. Gn. Bn for n = 1, …, 120; Table S1) and a
given plastic item i (Ri. Gi. Bi) are known, reference colour codes may be automatically
assigned by identifying the reference node at the minimum distance from Ri. Gi. Bi,
being the distance (D) between i and any reference node n estimated as:

𝐷!.! = 𝑅! − 𝑅! ! + 𝐺! − 𝐺! ! + 𝐵! − 𝐵! !

To compare results from digital and visual methods, we calculated the distance
between the RGB score obtained from the image and the reference node
corresponding to the colour assigned by the palette-assisted method for each of the
plastic items analysed with both methods (N = 103). The matching between methods
was deemed acceptable if this distance was smaller than 27. This distance implies that
the colour assigned by the digital method exactly matches with that resulting from the
S4
visual method, although occasionally the digital RGB score can lead to an adjacent
similar colour (with a closer node), especially among the light-colored plastics (Fig. S5).
Ninety-one percent of the colours assigned with the digital method matched with
those assigned from the visual method. Special attention should be taken in assigning
colours to light plastics including dark patches due to biofouling or shadows due to
irregular surface. Including these patches into the selection for colour averaging may
result in light grey colours.

Figure S5: Example of the distances between the RGB nodes in the 120-Palette and the
RGB score obtained from the digital image of an item classified as G7 by visual method.
In this case, the minimum distance was found for the G7 node, but other nearby colour
nodes may also show short distances (e.g. T8, C8, S8, B8). The shortest distances are
shown in red.

S5

Figure S6. Abundance of colours for the different plastic types (rigid fragment, film, foam,
fishing line, pellet and microbead). Scale bars in the pictures equate to 1 centimetre. Main
colours are shown in stacking bars, using single bars for dark, medium and light hues, and
black, white and transparent/translucent colours. Colour assignment was carried out by the
visual palette-assisted method. Data used for these figures are provided in Table S2.

S6

Figure S7: Percentages of colour for rigid fragments in relation to distance to land in open-
ocean (A; n = 3,446 items) and semi-enclosed regions (B; n = 5,403 items). Note that the
distance to land ranged from 26 to 2,737 km in open-ocean dataset, while the distance to land
ranged from 2 to 334 km in the semi-enclosed dataset. Percentages were calculated for
moving averages of 200 fragments. Single Value Decomposition (SVD) and Pearson correlation
coefficients were used to group colours showing similar covariance in relation to distance to
land and size. Figure legends show the colour groups. Colour assignment was carried out by
the visual palette-assisted method.

S7

Figure S8: Percentages of colour of rigid fragments in relation to plastic size for open-ocean (A;
n = 3,446 items) and semi-enclosed regions (B; n = 5,403 items). Percentages were calculated
for moving averages of 200 fragments. Single Value Decomposition (SVD) and Pearson
correlation coefficients were used to group colours showing similar covariance in relation to
distance to land and size. Figure legends show the main colour groups. Colour assignment was
carried out by the visual palette-assisted method.

S8

Figure S9. Handheld sample of plankton collected in a surface net tow in the Equatorial
Atlantic. Photo-protective pigmentation is commonly used for the zooplankton living on the
surface waters with high UV exposure. In this sample, copepods showed red and mostly blue
pigments. Photography by Unai Basurko de Miguel (PAKEA Bizkaia).

S9

Table S1: Colour codes. RGB scores and Pantone´s codes for the 120 reference colours
in 120-Palette (see Fig. 1). Digital RGB scores allow for locating references nodes in the
RGB colours space (Fig. S3), while Pantone´s codes allow for verifying a suitable
reproduction of the 120-Palette in printing format.

Colour Colour
R G B Pantone R G B Pantone
code code
Black 0 0 0 - V5 115 76 146 P91-15C
White 255 255 255 - V6 140 108 165 P91-14C
G1 18 21 23 Black 6C V7 169 144 189 P91-12C
G2 53 56 57 447 C V8 199 182 213 P91-11C
G3 86 87 87 425 C V9 227 218 235 P91-10C
G4 115 113 113 424 C M1 34 16 21 4975CP
G5 142 142 143 423 C M2 68 6 42 690CP
G6 164 165 165 422 C M3 100 22 64 235C
G7 187 186 187 Cool Grey 5C M4 128 21 81 P72-8C
G8 210 208 207 Cool Grey 3C M5 156 13 100 P71-16C
G9 231 230 228 427 CP M6 172 68 128 P71-15C
E1 25 37 20 5535 CP M7 191 113 156 P74-4C
E2 29 72 35 357CP M8 211 160 188 P73-11C
E3 45 106 52 349CP M9 233 208 221 P73-9C
E4 67 136 60 348CP P1 48 24 31 504CP
E5 87 160 64 P145-8C P2 92 49 61 506CP
E6 117 172 96 P145-6C P3 133 75 89 P70-13C
E7 147 187 131 7479 CP P4 174 99 117 P69-14C
E8 180 208 170 P145-3C P5 208 122 144 P68-5C
E9 217 232 212 P145-1C P6 216 148 165 P68-4C
T1 17 30 23 5605 CP P7 224 173 186 P68-2C
T2 10 64 48 3302CP P8 234 200 209 P73-2C
T3 10 95 72 568CP P9 243 226 231 P73-1C
T4 18 122 92 341CP R1 50 10 14 7631CP
T5 28 149 113 3278CP R2 88 20 21 1815CP
T6 74 161 136 3268CP R3 129 27 29 P58-16
T7 116 177 161 3265CP R4 164 29 34 P58-8C
T8 159 199 188 338CP R5 194 33 55 P55-8C
T9 205 226 221 331CP R6 200 87 83 P57-6C
C1 19 42 43 5463CP R7 210 125 123 P57-13C
C2 25 80 80 7476 CP R8 221 161 162 P55-11C
C3 41 116 115 P126-15C R9 238 211 208 P56-1C
C4 62 150 150 P127-8C O1 51 22 15 1545CP
C5 83 167 172 P124-7C O2 98 40 24 181CP
C6 114 179 183 P124-6C O3 143 62 29 P38-16C

S10
C7 146 194 199 P124-5C O4 187 85 24 P37-16C
C8 180 213 216 P124-11C O5 204 101 47 P37-8C
C9 216 233 235 P124-2C O6 211 128 75 P37-7C
S1 17 28 40 303CP O7 221 157 116 P37-5C
S2 10 57 78 302CP O8 230 187 161 P40-3C
S3 17 84 113 7700CP O9 241 220 207 P40-1C
S4 27 108 146 7705CP W1 32 22 14 7596CP
S5 41 131 176 314CP W2 65 44 28 7589 CP
S6 82 153 188 632CP W3 96 67 43 463C
S7 124 176 203 P121-13C W4 124 87 56 7575C
S8 167 201 218 P121-11C W5 153 108 71 P36-12C
S9 210 227 236 P118-10C W6 171 136 106 4725C
B1 15 18 35 296CP W7 192 165 143 480C
B2 11 35 69 282CP W8 213 195 180 4755C
B3 16 54 100 534C W9 233 225 216 7604C
B4 27 71 129 7687C Y1 51 52 17 7554CP
B5 41 88 156 7685C Y2 98 98 39 581CP
B6 87 119 174 P110-14C Y3 141 142 42 7747C
B7 128 151 192 P110-12C Y4 185 184 45 7765C
B8 171 186 212 P106-11C Y5 223 223 62 P163-8C
B9 212 220 232 P110-1C Y6 228 226 96 P163-7C
V1 26 15 34 532CP Y7 234 233 133 P163-6C
V2 50 28 67 518CP Y8 239 239 171 P163-4C
V3 73 43 97 525C Y9 246 247 212 P163-2C
V4 96 59 126 P92-7C Trans. - - - -

S11
Table S2: Number of plastic items classified according to colour and typology. Colours
correspond to a simplified version of the reference palette (see Methods and Fig. 2).

PLASTIC TYPE Total
Total
Rigid Fishing Micro- percentage
COLOURS Film Foam Pellet number
fragment line beads (%)
WHITE 2306 54 81 124 10 11 2586 29.2
TRANS. 1145 259 0 10 8 0 1422 16.1
(Transparent/translucent)
BLACK 492 37 1 61 13 10 614 6.9
GD – Grey dark 72 7 0 12 0 2 93 1.1
GM – Grey medium 193 9 0 37 1 4 244 2.8
GL – Grey light 131 8 0 27 0 0 166 1.9
RD – Red dark 2 0 0 0 0 0 2 0.0
RM – Red medium 33 4 0 3 1 0 41 0.5
RL – Red light 22 0 0 3 0 0 25 0.3
OD – Orange dark 2 0 0 0 0 0 2 0.0
OM – Orange medium 44 0 0 10 0 0 54 0.6
OL – Orange light 167 10 4 21 1 1 204 2.3
WD – Brown dark 41 1 0 0 1 0 43 0.5
WM – Brown medium 191 3 8 7 2 13 224 2.5
WL – Brown light 820 9 29 44 0 33 935 10.6
YD –Yellow dark 21 1 2 1 1 0 26 0.3
YM – Yellow medium 87 0 0 6 0 0 93 1.1
YL – Yellow light 733 15 65 46 13 2 874 9.9
ED – Green dark 12 2 0 6 0 0 20 0.2
EM – Green medium 16 1 0 4 0 0 21 0.2
EL – Green light 123 1 0 28 0 0 152 1.7
TD – Turquoise dark 14 1 0 0 0 0 15 0.2
TM – Turquoise medium 56 3 1 37 0 2 99 1.1
TL – Turquoise light 160 7 1 111 0 1 280 3.2
CD – Cyan dark 7 0 0 1 0 0 8 0.1
CM – Cyan medium 107 4 0 20 0 0 131 1.5
CL – Cyan light 113 10 0 18 0 0 141 1.6
SD – Sky dark 9 1 0 2 0 0 12 0.1
SM – Sky medium 106 6 0 19 0 0 131 1.5
SL – Sky light 64 2 0 11 0 0 77 0.9
BD – Blue dark 2 0 0 2 0 0 4 0.0
BM – Blue medium 11 0 0 2 0 0 13 0.1
BL – Blue light 29 1 0 0 0 0 30 0.3
MD – Magenta dark 1 0 0 0 0 0 1 0.0
MM – Magenta medium 24 1 0 2 0 0 27 0.3
ML – Magenta light 39 0 0 0 0 0 39 0.4
Total number 7395 457 192 675 51 79 8849 100
Total percentage (%) 84 5 2 8 1 1 100

S12
Table S3: Percentages of colors of rigid fragments in the 120-Palette. A total of 7,395
items were included in the present analysis. Data are graphed in Fig. 3.
S13

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Contaminants in Aquatic and Terrestrial Environments

is published by the American Chemical Society. 1155 Sixteenth Street N.W.,

The colours of the ocean plastics

2 Characterisation of the colour is often included in studies on plastic pollution. However,

3 the comparability and relevance of this information is limited by methodology or

4 observer subjectivity. Based on the analysis of thousands of floating plastic fragments

5 from a global collection, here we propose a systematic semi-automatic method to

14 order modulation of the colour distributions of marine plastic microplastics by the

17 colour information on ocean plastics.

19 Table of Contents (TOC) Art

23 to a progressive degradation of the chemical bond structure of the polymers (Brandon et

27 2017; Cózar et al. 2017).

29 environment. The stages of weathering or the time scales of degradation, fragmentation

32 undergoes photo-oxidation and changes in the absorbance and reflection of light,

42 classifications based on up to 32 colours (Blair Crawford & Quinn 2017). There is a

46 subjective and depends, among others, on visual capabilities and experience of

47 observer. The lack of standardized methods imposes significant difficulties to compare

48 studies and achieve robust conclusions.

51 analysis to define a consistent methodological tool to characterize colour in marine

54 on sources and fate of marine plastics. Based on weathering experiments (Andrady

55 2017), we expect photo-oxidative damage to lead to increasing fractions of white and

56 yellow-brown colours in ocean plastics with increasing exposure time in the

58 transport of plastic fragments to distant open-ocean accumulations is proposed to be a

63 and dynamics of marine plastic litter.

64 Material and Methods

69 Atlantic and Indian Ocean) as well as semi-enclosed regions (Mediterranean, Arctic

76 2018). Other major commodity plastics such as polystyrene (PS), polyethylene

79 Table 1. Summary of sampling regions, date and number of plastic items (N =

Region/Cruise Date No. Items Ref.

87 selected a 120-colours scheme in order to find a balanced equilibrium between

92 proposed to the palette-assisted assignment of colours, a visual method and a digital

93 semi-automatic method, both providing comparable results (see Supporting Methods).

103 medium and light hues (42-Palette, see Methods).

105 Visual palette-assisted method for colour assignment

113 analyses were exclusively based on the primary colour.

114 Digital semi-automatic method for colour assignment

128 score by proximity in the colour space (see Supporting Methods).

129 Size and shape of the plastic items

140 dataset) as wide magenta.

152 main eigenfunctions were represented jointly as “Rest of colours”.

162 and green colours (Fig. 2, Fig. S6).

169 provided in Table S2 and graphed in bar charts in Fig. S6.

188 transparent/translucent and black (31 %, 16 % and 7 %, respectively). Colour

190 percentages for each colour are provided in Table S3.

204 landward, contrary to the white colour.

213 in the load of white fragments.

232 types on colour distributions (Fig. 2).

234 and colour of the plastic polymers. Comparing recently-manufactured objects,

239 fragmentation. In parallel, photo-oxidation generally causes changes in plastic colour,

241 light colours (discolouration or whitening), while the accumulation of degradation

243 colours (yellowing and tanning, respectively) (Andrady 2017).

257 was relatively low.

272 enrichment in blue microplastics on the ocean surface. In lakes without

273 zooplanktivorous pressure, a more cost-effective red pigmentation is used by the

274 zooplankton to protect themselves against UV radiation; however, UV-protective

276 zooplanktivorous fishes increase (Hudelson 2011). Therefore, if the removal of

294 out yet in relation to the abundance of bluish plastics at sea.

315 resins in the market.

324 Andrady 2017).

339 developed a comparable alternative digital method to advance in the automatization of

340 the colour assessment if future analyses (see Supporting Methods).

352 ASSOCIATED CONTENT