Science of the Total Environment 675 (2019) 686–693

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Science of the Total Environment

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

Release of micro- and nanoparticles from biodegradable plastic during in

situ composting
Henry Y. Sintim a,b, Andy I. Bary a,b, Douglas G. Hayes c, Marie E. English c, Sean M. Schaeffer c, Carol A. Miles d,
Alla Zelenyuk e, Kaitlyn Suski e, Markus Flury a,b,⁎
a
Department of Crop & Soil Sciences, Washington State University, Pullman, WA 99164, United States of America
b
Department of Crop & Soil Sciences, Washington State University, Puyallup, WA 98371, United States of America
c
Department of Biosystems Engineering and Soil Science, University of Tennessee, Knoxville, TN 37996, United States of America
d
Department of Horticulture, Washington State University, WSU Mount Vernon, Northwestern Washington Research & Extension Center, Mount Vernon, WA 98273, United States of America
e
Pacific Northwest National Laboratory, Richland, WA 99354, United States of America

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

• Plastic pollution is a ubiquitous environ-

mental problem.
• Biodegradable plastic can help reduce
plastic pollution.
• Not all components in biodegradable
plastics may degrade.
• Non-biodegradable additives, such as
carbon black, may be released upon
degradation of plastic.

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

Article history: Plastic is ubiquitous in modern life, but most conventional plastic is non-biodegradable and accumulates as waste
Received 16 February 2019 after use. Biodegradable plastic is a promising alternative to conventional plastic. However, biodegradable plas-
Received in revised form 9 April 2019 tics must be thoroughly evaluated to ensure that they undergo complete degradation and have no adverse impact
Accepted 11 April 2019
on the environment. We evaluated the degradation of biodegradable plastics during 18-week full-scale
Available online 18 April 2019
composting, and determined whether additives from the plastics are released upon degradation. Two biodegrad-
Editor: Jay Gan able plastic films—one containing polybutylene co-adipate co-terephthalate (PBAT) and the other containing
polylactic acid/poly-hydroxy−alkanoate (PLA/PHA)—were placed into meshbags and buried in the compost.
Keywords: Degradation was assessed by image analysis, scanning electron microscopy, Fourier-transformed infrared spec-
Biodegradable plastic troscopy, electrophoretic mobility, δ13C isotope analyses, and single particle mass spectrometry of mulch frag-
ASTM D6400 ments. The results showed N99% macroscopic degradation of PLA/PHA and 97% for PBAT film. Polymers in the
ASTM D5338 biodegradable films degraded; however, micro- and nanoparticles, most likely carbon black, were observed on
Mulch the meshbags. Overall, biodegradable plastics hold promise, but the release of micro- and nanoparticles from bio-
Compost
degradable plastic upon degradation warrants additional investigation and calls for longer field testing to ensure
Microparticles
that either complete biodegradation occurs or that no long-term harm to the environment is caused.
Nanoparticles
Degradation © 2019 Elsevier B.V. All rights reserved.

⁎ Corresponding author at: Department of Crop & Soil Sciences, Washington State University, Puyallup, WA 98371, United States of America.
E-mail address: fl[email protected] (M. Flury).

https://doi.org/10.1016/j.scitotenv.2019.04.179
0048-9697/© 2019 Elsevier B.V. All rights reserved.
 H.Y. Sintim et al. / Science of the Total Environment 675 (2019) 686–693 687

1. Introduction 2. Materials and methods

Plastic pollution of terrestrial and aquatic ecosystems is an omni- 2.1. Experimental approach
present and ever-growing threat to the environment. On average,
every person in western Europe and the US consumes 100 kg of plas- Two biodegradable plastic mulch films and one conventional non-
tic products every year, while in Asia this number is 20 kg, and rising biodegradable polyethylene mulch film were used under normal agri-
quickly (Gourmelon, 2015). Global production of plastic in 2015 was cultural practices for one season of pumpkin production. Mulch films
400 million metric tons, but 300 million metric tons of plastic waste were in the field for 113 days and removed after pumpkin harvest.
was generated (Geyer et al., 2017). Only a small fraction (9 to 26%) of Mulch film pieces were then placed into meshbags that were buried
used plastic is recycled, with 22 to 43% being discarded in landfills in a compost pile to assess their degradation during composting. Degra-
(Gourmelon, 2015). It is projected that by 2050, globally 9 billion dation was quantified by image analysis, scanning electron microscopy,
metric tons will have been recycled, 12 billion metric tons inciner- electrophoretic mobility, organic carbon analysis, FTIR, and Scanning
ated, and 12 billion metric tons discarded in landfills or the natural Mobility Particle Sizer and single particle mass spectrometry
environment (Geyer et al., 2017). Most of the plastics produced (miniSPLAT).
worldwide are non-biodegradable, which has the undesired conse-
quence that plastics inadvertently released into the environment
2.2. Plastic mulch films
will accumulate over time.
The increasing use of plastics and the associated waste that is gener-
Three plastic mulch films were used: two biodegradable mulches
ated cause enormous environmental and economic challenges. Proper
(BioAgri® and PLA/PHA) and a conventional polyethylene mulch,
disposal of plastics has not been achieved as shown by the plastic pollu-
which was used as a control. The two biodegradable plastic films repre-
tion of the oceans, which are often the final receptors of plastic waste
sent two typical classes of biodegradable polymers used by the industry:
(Worm et al., 2017; Haward, 2018). Efforts are underway to find ways
(1) a co-polyester containing PBAT, and (2) polylactic acid (PLA)-poly
to degrade conventional plastics (Yang et al., 2014; Yang et al., 2015;
(hydroxy−alkanoate) (PHA) blend, respectively. These plastics are typ-
Bombelli et al., 2017), and initial results of using both macroorganisms
ically used as mulch films in agricultural applications. The PBAT contain-
(Yang et al., 2014; Yang et al., 2015; Bombelli et al., 2017; Yoshida
ing plastic film is a commercial product called BioAgri®, whose key
et al., 2016) and microorganisms (Yoshida et al., 2016) to biodegrade
constituent is Mater-Bi®; the PLA/PHA film is an experimental product.
conventional plastics have been promising, although some of the find-
All the plastics were black, due to the presence of carbon black, com-
ings have been questioned (Drahl, 2018; Weber et al., 2017). A promis-
monly used as an additive for color and enhanced performance and du-
ing alternative to conventional plastics is biodegradable plastics.
rability. For instance, it has been demonstrated that the presence of
Biodegradable plastics can be converted by microorganisms into carbon
carbon black helps to retain the mechanical mulch properties of PLA/
dioxide, microbial biomass, and water (Brodhagen et al., 2015;
PHA films during weathering (Mosnackova et al., 2017). Specifics
Zumstein et al., 2018).
about manufacturers and composition of the mulches are given in the
The use of biodegradable plastic will reduce disposal needs and the
Supporting Information “Plastic Mulch Films” and Tables S1 and S2.
negative environmental impacts associated with conventional plastics,
The two biodegradable plastic films used here are known to biode-
but it has to be demonstrated that they indeed completely degrade ei-
grade in compost as evaluated with ASTM D6400 (ASTM, 2012),
ther in the environment or during composting (Sintim and Flury,
which specifies that biodegradation is demonstrated when after
2017). Composting, in particular, may be a viable means of disposal
180 days N90% of the organic carbon of the entire plastic or of each or-
for biodegradable plastics, particularly in climates where complete deg-
ganic constituent, which is present in the material at N1%, has been con-
radation in soil does not occur (Calabr and Grosso, 2018; Hottle et al.,
verted to carbon dioxide when tested with ASTM D5338 (ASTM
2013; McGill and Oshins, 2018).
International, 2015). The polymers in the biodegradable plastics used
Several biodegradable polymers have been developed and tested
in our study satisfy this biodegradation criterion (BioBag, 2018;
for biodegradability under a composting environment. For instance,
Greene, 2018).
poly(butylene adipate-co-terephthalate) (PBAT), an important co-
Detailed mechanistic studies on biodegradation, including CO2 re-
polyester used for the production of biodegradable plastics, can be
lease and change in mechanical and molecular properties of PBAT and
metabolized into carbon dioxide, water, and microbial biomass in
PLA/PHA films, have been reported elsewhere (Kijchavengkul et al.,
compost (Kijchavengkul et al., 2010). Standard biodegradation
2010; Mosnackova et al., 2017). However, what is not known is whether
tests for plastics in compost, such as the ASTM D6400 (ASTM,
additives, such as carbon black, are released during the degradation of
2012) and ASTM D5338 (ASTM International, 2015), consider a plas-
plastic polymers. In laboratory degradation tests, such as ASTM D5338
tic to be biodegradable when during 180 days of composting N90% of
(ASTM International, 2015), the primary constituents/ingredients are
the organic carbon of the entire plastic, or each organic constituents
ground to a powder and assayed, so these tests do not necessarily assess
present in quantities N1%, are converted into CO2 when compared to
the degradation of the final plastic product.
a positive control. The test specimens can be in the form of films,
granules, or powder (ASTM International, 2015), while constituents
present in the plastic b1% in mass do not need to be tested (ASTM, 2.3. Field experiment
2012).
However, it still needs to be demonstrated that biodegradable plas- A field experiment with Cucurbita pepo L. (pumpkin) was carried out
tics completely degrade in situ in the natural or composting environ- at Mount Vernon, WA. Details are given in the Supporting Information
ment. It is also not known to what degree minor components, such as “Field Experiment”. After pumpkin harvest, mulch pieces (10 cm ×
carbon black, used in the production of biodegradable plastics, will be 12 cm) were transferred into clear nylon meshbags (mesh opening
released upon degradation of biodegradable plastic and whether they 250 μm), which were then placed into a compost pile to investigate
will undergo biodegradation. Thus, our objectives were to (a) evaluate mulch degradation (see Supporting Information “Processing of Field-
the degradation of biodegradable plastic in situ in compost and Weathered Mulch Films and Meshbags”, Fig. S1). Exposure of plastic
(b) determine whether particulates that are added in the production mulches to UV radiation can either lead to cross-linking and thereby a
of plastics are released upon degradation. This is the first time that par- reduction of biodegradability (Kijchavengkul et al., 2008), or alterna-
ticle release from biodegradable mulch under real composting condi- tively weaken the mechanical properties of plastic films through depo-
tions is reported. lymerization (Hayes et al., 2017), thereby promoting biodegradation.
688 H.Y. Sintim et al. / Science of the Total Environment 675 (2019) 686–693

2.4. Composting 2.8. Scanning Mobility Particle Sizer (SMPS) and single particle mass spec-
trometry (miniSPLAT)
The compost was an aerated static pile, consisting of dairy manure
solids (28% vol.), broiler litter (28%), yard wastes (28%), animal bedding Scanning Mobility Particle Sizer and single particle mass spectrome-
(14%), and fish carcasses (2%), set up at Washington State University's try (miniSPLAT) were conducted to determine size distributions and
Puyallup Research & Extension Center, Puyallup, WA. The feedstock particle mass spectra of the residues remaining on the meshbags. Single
was formulated to provide a C:N ratio of 25–30:1 and 55–65% gravimet- particle mass spectrometer, miniSPLAT was used to characterize vac-
ric moisture content, the recommended conditions to generate suffi- uum aerodynamic diameter and composition of individual particles.
cient heat that will meet the requirements for pathogen reduction The mass spectra of particles produced from aerosolizing all the samples
(U.S. Composting Council, 2001; USEPA, 1999). The compost pile was were classified into classes or clusters based on the similarities in their
≈5-m wide × 9-m long × 2-m high, and was under a roof cover to pro- mass spectra using SpectraMiner software (Zelenyuk et al., 2006) and
tect it from rain. displayed using interactive circular hierarchical tree or dendrogram.
The meshbags were placed at 60-cm depth in the compost pile (at For more details, see the Supporting Information “Scanning Mobility
the time of pile construction). The meshbags were retrieved at 2-week Particle Sizer and Single Particle Mass Spectrometry”.
intervals for the first seven sampling times, and the eighth sampling
time was after 18 weeks of composting. The pile temperature at 60- 2.9. Stable isotope analysis (δ13C)
cm depth was monitored continuously at different locations within
the compost pile using temperature sensors and data loggers (TMC50- Stable isotope analysis was conducted on mulch residues dislodged
HD sensors and U12-008 data loggers, Onset Computer Corp., Bourne, from the meshbag fibers. Pieces of the nylon meshbags were soaked in
MA). Air temperature was monitored at the Washington State Univer- deionized water for 24 h to dislodge the residues. The dried residues
sity AgWeatherNet Station in Puyallup, located 300 m away from the were then analyzed for their 13C/12C ratio. For more details, see the
compost pile. Supporting Information “Stable Isotope Analysis”.

2.5. Macroscopic surface area 2.10. Carbon black

After retrieval from the compost, the meshbags were transferred to a Carbon black, the same batch that was used for the production of the
laboratory, cleaned gently to remove adhering compost, air-dried, and PLA/PHA mulch film, was obtained from Techmer PM Polymer Modi-
then photographed to provide a visual configuration of mulch degrada- fiers (Clinton, TN). This carbon black was also analyzed for electropho-
tion. The meshbags were then cut open and the mulch pieces trans- retic mobility, FTIR, δ13C, and SMPS and miniSPLAT.
ferred onto a white paper. A non-serrated sandwich spreader was
used to transfer mulch fragments adhered to the meshbags onto the 2.11. Statistical analysis
white paper. The mulch fragments were spread uniformly on the
white surface and photographed. ImageJ software (Rasband, 2014) Statistical analyses were performed with ANOVA in R (Version 3.3.2)
was used to digitize the photographs and to measure the number of par- (R Core Team, 2016). Significance and mean comparisons (performed
ticles, total area, and total perimeter of the degraded mulch samples. with the Least Significant Difference (LSD) comparison) were deter-
The total area was expressed as a percentage of the initial surface area mined at P = 0.05 and P = 0.1.
of the buried mulch samples.
3. Results

2.6. Scanning Electron microscopy and electrophoretic mobility 3.1. Macroscopic mulch degradation

Meshbag fibers and mulch fragments were examined by scanning The biodegradable plastic mulches visually degraded during
electron microscopy (SEM FEI Quanta 200F SEM; GenTech Scientific composting, whereas the polyethylene did not (Fig. 1A). Despite signif-
Inc., Arcade, NY). No particles were visible on clean, non-used icant macroscopic degradation of the biodegradable plastic films, black
meshbags. For composted meshbags, however, we could visually and stains were observed on the meshbags used to contain the plastic
microscopically see mulch residues adhered to the nylon fibers of the films (Fig. 1A). The stains were more apparent in the meshbags that
meshbags. These residues were analyzed for electrophoretic mobility, contained PLA/PHA than those that contained BioAgri®, but no stains
FTIR, and δ13C. For electrophoretic mobility, meshbags were placed were found in the meshbags that contained polyethylene and no-
into deionized water and soaked overnight. This caused the residues plastic controls. These stains were caused by residues from the biode-
to dislodge from the fibers. The pH was adjusted to about pH 7 and gradable plastic films. The stains were no longer identifiable by the
the ionic strength to 1 mM NaNO3. The solution was then decanted, son- naked eye after 18 weeks of burial, because the meshbags were then
icated for 1 min, and analyzed for electrophoretic mobility with a strongly darkened with compost material due to the prolonged expo-
ZetaSizer Nano S (Malvern Instruments, Malvern, UK). sure to compost.
Fig. 1B shows the quantification of the macroscopic film degradation
2.7. Fourier-transformed infrared spectroscopy (i.e., loss of area%) and the temperatures during composting. Both bio-
degradable plastics showed rapid macroscopic degradation (Fig. 1B):
Mulch residues were further analyzed with Attenuated Total Reflec- PLA/PHA showed 92% loss in surface area within the first two weeks,
tance (ATR)-FTIR Spectroscopy (IRAffinity-1, Shimadzu Co., Tokyo, whereas BioAgri® showed a 60% loss. After 18 weeks of composting
Japan; MIRacle ATR, PIKE Technologies, Madison, WI, USA). We could the loss of surface area was N99% for PLA/PHA and 97% for BioAgri®.
not obtain sufficient mulch material for FTIR by scraping of mulch As expected, there was no degradation of polyethylene even after
from the meshbag fibers, so we cutoff meshbag pieces of 1 cm × 1 cm 18 weeks of composting.
that contained mulch residues, and then analyzed these pieces with
FTIR. Clean meshbags were also analyzed to obtain the background 3.2. Microscopic mulch degradation
spectrum for the nylon. Measurement resolution was 2 cm−1, and 16
scans per spectrum were recorded. Peaks were assigned following Scanning electron micrographs showed no evidence of particles ad-
Hayes et al. (2017). hered to clean, non-used meshbags, but that there were residues of the
 H.Y. Sintim et al. / Science of the Total Environment 675 (2019) 686–693 689

A Initial Week 4 Week 8 Week 12 Week 18

BioAgri A B

BioAgri
PLA/PHA

500µm 50µm

C D

PLA/PHA
Polyethylene

500µm 50µm

B E F

Polyethylene
Temperature (°C) Mulch surface area remaining (%)

100
BioAgri
PLA/PHA
75 Polyethylene

50 500µm 50µm

25 G H

Unused
0
80 Atmosphere
60 Compost
40
20
500µm 50µm
0
0 4 8 12 16 Fig. 2. Scanning electron micrographs of meshbag nylon fibers after 18 weeks of
Time (week) composting. Images show meshbag fibers at two different magnifications. (A,
B) BioAgri®, (C, D) PLA/PHA, (E, F) polyethylene, (G, H) unused meshbag.
Fig. 1. Macroscopic degradation of plastic films in compost. (A) Visual degradation of
plastic films in meshbags after composting for different times. Plastic films were
enclosed in nylon meshbags for composting. Size of initial plastic pieces is 10 cm × functional groups of compost materials with time. The particles in the
12 cm, as shown as dark rectangles. The round token is the aluminum label used for meshbag that contained BioAgri® and PLA/PHA were also negatively
identification. The extensive dark staining at week 18 is caused by the compost itself.
charged. After 4 weeks of composting, the electrophoretic mobility of
(B) Percentage of plastic film surface area remaining at different stages of composting,
and atmospheric and compost temperatures during composting. Error bars (n = 4) and BioAgri® and PLA/PHA were significantly different (P = 0.014), but nei-
gray ribbon (n = 8 at initial date, but reduced by 1 at each subsequent sampling date) ther of the two was different from carbon black and compost. After
represent the standard deviation of the mean. 18 weeks, the electrophoretic mobility of both PLA/PHA and BioAgri®
particles approached that of carbon black, whereas the compost (from
plastic films on the meshbag nylon fibers after 18 weeks of composting the polyethylene meshbags) had more negative electrophoretic mobil-
(Fig. 2). Residues were observed on the meshbags of all three films; ity. Differences, however, were not statistically significant (P = 0.110
however, those found on the meshbag that contained polyethylene to 0.150).
were much less, and clearly distinct in shape and structure from those Fig. 3B shows the δ13C values for the initial plastic films (0 weeks)
on the meshbags that contained BioAgri® and PLA/PHA films. BioAgri® and the film residues at different times during composting. The δ13C of
meshbags appear to contain plastic fragments, whereas the PLA/PHA field-weathered polyethylene (0 weeks) is −29.9(%). The original
meshbags contained much smaller particles that were not recognizable PLA/PHA had a significantly less negative δ13C than BioAgri®, carbon
as plastic fragments. Higher magnification micrographs suggest that the black, and compost (P b 0.001). This is because PLA/PHA is made of
residues are micro- and nanoparticles that formed aggregates (Fig. 2B, corn feedstock (C4 carbon). During composting, however, the -δ13C of
D). Even micrographs of the unburied films indicate the presence of par- the PLA/PHA residues more and more approached the values of
ticulates (Supporting Information, Fig. S2). BioAgri®, polyethylene, and carbon black. At week 18, the δ13C of
The black color of these residues suggests the presence of carbon BioAgri® (−27.1%) and PLA/PHA (−27.1%) were not significantly dif-
black. The carbon black pigment used for preparing PLA/PHA consisted ferent, but their δ13C were significantly more negative than that of poly-
of a polydisperse population of particles, and some particles formed ag- ethylene (−26.8 % ), but less negative than that of carbon black
gregates (Supporting Information, Fig. S3A). The black residues on the (−27.7%). This suggests that the corn-based components of the PLA/
meshbags that contained BioAgri® and PLA/PHA after 18 weeks of PHA plastic were degrading, leaving behind a non-corn based residue,
composting also possessed a polydisperse size distribution, but they which was different from the compost.
were bigger than the pure carbon black component of PLA/PHA. The FTIR spectra of recovered meshbags indicate that the polymers
of the biodegradable plastics degraded with time (Fig. 3C). The spectra
3.3. Particle characterization reflect the signal from the nylon meshbag, which did not degrade. Char-
acteristic peaks for the biodegradable plastics that do not overlap with
The particles on the nylon fibers of the meshbags that contained nylon are at 1750–1700 cm−1 (carbonyl stretch) and
polyethylene were negatively charged, and their electrophoretic mobil- 1150–1000 cm−1 (C\\O stretch). For BioAgri®, the 1750–1700 cm−1
ity became more negative with composting time (weeks 4, 8, and 18) peak decreased with time, but was still present after 18 weeks of
(Fig. 3A). As the polyethylene did not degrade, we conclude that the composting (Fig. 3C). This indicates that the plastic is degrading, but
particles from the polyethylene meshbags stem from the compost itself. has not fully decomposed at the end of composting. This is supported
The decrease in electrophoretic mobility suggests an increase in by the microscopic observations that showed possible plastic fragments
690 H.Y. Sintim et al. / Science of the Total Environment 675 (2019) 686–693

Electrophoretic mobility
BioAgri C (a) BioAgri Carbon black
A PLA/PHA 8 Unused meshbag

(μm s−1/ V cm−1)

Absorbance
Initial mulch
Polyethylene Week 4
−1 Carbon black 6
Week 8
Week 18
4
−2
2

−3 0
0
B (b) PLA/PHA
8

Absorbance
−10
6
δ13C (‰)

−20 4

−30 2

0
0 4 8 12 16 20
Time (Week) (c) Polyethylene
8

Absorbance
6

0
1800 1600 1400 1200 1000
Wavenumber (cm−1)

Fig. 3. Physico-chemical characterization of plastic residues. (A) Electrophoretic mobility and (B) δ13C of plastic residues collected from meshbag nylon fibers as a function of composting
time. Pure carbon black was not composted and is indicated as a horizontal line. Measurements of electrophoretic mobility were made at pH 6.7 to 7.1 and ionic strength of 1 mM NaNO3.
The black box indicates values determined from the initial field-weathered plastic mulches. (fC) FTIR spectra of meshbag fibers with adhered plastic: BioAgri®, PLA/PHA, and polyethylene
films at different times during composting. Black lines indicate spectra of pure carbon black nanoparticles (dashed lines) and unused nylon meshbag (dotted lines); colored lines indicate
spectra of plastic films.

Fig. 4. Particle size distribution and Single Particle Mass Spectrometry (miniSPLAT). (A, B) Particle mobility size distribution of carbon black and residues on meshbags after week 8 and 18
of composting. Data represent means (lines) and standard deviations (bands) of three replicates. Data for carbon black are multiplied by a factor of 5 to make them more visible.
(C) Dendrograms of cluster analysis of carbon black particles and particles adhered to meshbags after week 8 and 18of composting. Each line represents a type of particle, and the
color indicates the fraction of particles of that composition. The red lines indicate many particles and blue very few. Black lines indicate zero particles. Numbers are percentages in
branches.
 H.Y. Sintim et al. / Science of the Total Environment 675 (2019) 686–693 691

on the meshbag fibers (Fig. 2A, B). For PLA/PHA, the C_O stretch de- all fragments would pass a 2-mm sieve. The two plastics are also certi-
creased with time and completely disappeared at 18 weeks (Fig. 3C), in- fied biodegradable in compost (N90% conversion of organic carbon
dicating complete degradation of the plastic polymers. This is again into CO2) according to the ASTM D5338 test (BioBag, 2018; Greene,
supported by the microscopic images (Fig. 2C, D). Similar patterns can 2018).
be seen in the 1150–1000 cm−1 absorbance region. Carbon black, due
to its dominantly pure elemental carbon structure, does not contribute 4.2. Carbon black
any FTIR peaks (Fig. 3C). FTIR spectra for polyethylene did not change
(Fig. 3C), supporting the hypothesis that polyethylene did not degrade. The remaining residues of the biodegradable plastics after 18 weeks
Size distributions acquired by the scanning mobility particle sizer of composting likely contain carbon black. Multiple evidence points to
(SMPS) showed that the residues on the meshbags contained the presence of carbon black: (1) While the interior of the nylon mesh
nanometer-sized particles (Fig. 4A, B). The distribution of pure carbon was completely empty, the nylon fibers were stained black, leaving be-
black particles peaked at a mobility diameter of 60 nm, BioAgri® at hind a “shadow” of the original mulch film piece (Fig. 1). The only agent
around 90–100 nm, and PLA/PHA and polyethylene (compost) both at that renders black color in the plastics is carbon black; (2) Scanning
60 nm. The mass spectrometry measurements by single particle mass electron micrographs showed that the black stain was of particulate na-
spectrometer, miniSPLAT, are indicative of predominantly carbon- and ture (Fig. 2); (3) Electrophoretic mobility of these particles was similar
oxygen-containing particles (Supporting Information, Fig. S4). The to that of the pure carbon black used in the production of the plastics;
data point to the presence of particles of different sizes, shapes, and (4) δ13C values of the particles were similar to that of carbon black;
compositions. We classified the particles into different classes or clus- and (5) FTIR showed successive degradation of plastic polymers during
ters based on the similarities in their mass spectra and quantified for composting, with almost complete degradation after 18 weeks of
each sample the relative fractions of particles in each cluster composting. A definitive identification, however, was not possible be-
(Zelenyuk et al., 2006). This analysis indicates that the residues on the cause of background noise from compost particles.
meshbags were all distinctly different from each other, but also different Carbon black is a type of elemental carbon produced mainly by
from pure carbon black (Fig. 4C). With increasing composting time, the vapor-phase pyrolysis of hydrocarbons under controlled conditions
particles originating from compost (indicated by the polyethylene clus- (IARC, 2010b; Long et al., 2013). It is classified as a carcinogen in exper-
ters) became more dominant, consistent with the more pronounced imental animals and as a Group 2B possible carcinogen to humans by
staining of the meshbags (Fig. 1). the International Agency for Research on Cancer (IARC, 2010a; IARC,
Taken together, the electrophoretic mobility, FTIR, SMPS, and 2010b). The National Institute for Occupational Safety and Health
miniSPLAT analyses suggest that the two biodegradable plastics de- (NIOSH) recommends treating carbon black as a potential human car-
graded, but micro- and nanometer sized particles remained on the cinogen when its concentration of PAH's is N0.1% (U.S. Department of
nylon fibers. These particles could not unequivocally be identified as Health and Human Services, 1988). Carbon black was found not to pres-
pure carbon black; however, based on their color, chemical, and physi- ent a risk to the environment or human health by EPA (Environmental
cal characteristics, they likely contained carbon black. Carbon black is Protection Agency, 2009) and Environment Canada (Environment
expected to undergo chemical and physical transformations, Canada, 2013). Like other carbonaceous materials, carbon black can be
e.g., adsorb various species that are present in the environment and un- a sink for organic pollutants in soil (Fitch and Smith, 1979; Prete et al.,
dergo surface reactions like oxidation, and this is why we could not un- 2017; Skinner and Beattie, 1916); but overall, little is known about the
equivocally identify the particles. fate of carbon black in terrestrial ecosystems and its toxicity to soil
micro- and macroorganisms. Carbon black is considered to be non-
4. Discussion biodegradable (Environment Canada, 2013); thus, if the polymers in
biodegradable plastic films will ultimately completely degrade, carbon
4.1. Degradation into micro- and nanoparticles black will be released.

The biodegradable plastic films studied here underwent different 4.3. Limitations
degradation in compost, which can be attributed to the type of source
material and the physicochemical properties of the plastics. BioAgri® The particles observed on the meshbags in this study were detected
is made of N-type Mater-Bi®, which consists of polybutylene co- because the white nylon meshbags contrasted with the dark particles
adipate co-terephthalate (PBAT) (Borchani et al., 2015). PBAT is re- and also by the change of FTIR spectra with time. Scanning electron mi-
ported to be readily degradable by bacteria in compost (Müller et al., croscopy revealed that particles from the plastics were adhering to the
2001; Witt et al., 2001), but less so by fungi (Saadi et al., 2013). PLA/ nylon fibers. Particles b250 μm may have not necessarily degraded,
PHA mulch film is made from a blend of polylactic acid (PLA) and poly but rather may have fallen through the meshbag, except for those that
(hydroxy−alkanoate) (PHA), both of which are biobased polymers. adhered to the nylon fibers. This would lead to an underestimation of
PLA has high glass transition temperature (≈60 °C) and a limited num- the particles observed in our study. The fragmentation of the biodegrad-
ber of microbial degraders; hence does not degrade readily in soil able plastics, as observed in the first 8 weeks of composting, produced
(Hayes et al., 2012; Mosnackova et al., 2017; Suyama et al., 1998; thousands of macro and microscopic particles from a 10 cm × 12 cm
Tokiwa et al., 2009). Studies, however, show that PLA undergoes satis- plastic piece (Table S3), and even more nanosized particles as detected
factory degradation in compost (Hayes et al., 2012; Rudnik and by SMPS and miniSPLAT mass spectrometry. Some of these particles
Briassoulis, 2011). In contrast to PLA, PHA is readily degradable under may have further degraded in the compost after 18 weeks.
most environmental conditions (Rudnik and Briassoulis, 2011; It is likely that the 250 μm nylon meshbags used in our study slowed
Vroman and Tighzert, 2009). the degradation of the plastics because some of the plastic was covered
International standards for satisfactory disintegration of plastics in by the nylon fibers (see Supporting Information “Effect of Meshbags on
compost (e.g., ASTM D6400 and ISO 17088) specify that after Mulch Film Degradation”). Biodegradation of plastics occurs at the sur-
12 weeks of composting, b10% of the original weight of plastics must re- face; thus the presence of a non-biodegradable cover can affect the deg-
main upon washing the composted materials in water and fractionating radation rate. Nonetheless, without a protective net, the retrieval of
on a 2-mm sieve (ASTM, 2012; ISO, 2012). By this criterion, the two fragments and particles becomes virtually impossible because frag-
studied biodegradable plastic mulch films underwent satisfactory disin- ments and particles possibly released by the plastic film would become
tegration, because after 12 months of composting, all the fragments left entangled with the surrounding compost. The use of mesh bags can
in the meshbags were b2 mm in size as observed microscopically, and serve as a potential source of experimental artifacts. Meshbags may
692 H.Y. Sintim et al. / Science of the Total Environment 675 (2019) 686–693

have decreased the degradation rate, and thus the particles detected at Borchani, K.E., Carrot, C., Jaziri, M., 2015. Biocomposites of alfa fibers dispersed in the
Mater-Bi type bioplastic: morphology, mechanical and thermal properties. Compos.
the nylon fibers could have completely degraded in the absence of a Part A 78, 371–379.
shielding net. While this may likely be the case for the residual polymers Brodhagen, M., Peyron, M., Miles, C., Inglis, D.A., 2015. Biodegradable plastic agricultural
detected for the BioAgri® plastic, we presume that carbon black nano- mulches and key features of microbial degradation. Appl. Microbiol. Biotechnol. 99,
1039–1056.
particles will be released even in the absence of a meshbag. Calabr, P.S., Grosso, M., 2018. Bioplastics and waste management. Waste Manag. 78,
800–801.
5. Implications Drahl, C., 2018. The truth about plastic eaters. Headlines about plastic-munching mi-
crobes and worms belie tough, competitive road to recycling processes. C&EN
(June), 30–33.
When biodegradable plastics are composted, and the plastic poly- Environment Canada, 2013. Screening Assessment for the Challenge: Carbon Black. Envi-
mers degrade, minor additives, such as carbon black are released from ronment Canada, Health Canada, Ottawa, Canada.
Environmental Protection Agency, 2009. Carbon black; exemption from the requirement
the plastic. The amounts of such additives in plastic films are not always of a tolerance. Fed. Regist. 74, 40509–40513.
known. If these additives do not biodegrade, the repeated application of Fitch, W.L., Smith, D.H., 1979. Analysis of adsorption properties and adsorbed species on
biodegradable plastic mulch films can thus lead to accumulation (not commercial polymeric carbon. Environ. Sci. Technol. 13, 341–346.
Geyer, R., Jambeck, J.R., Law, K.L., 2017. Production, use, and fate of all plastics ever made.
necessarily in mass, but in number) of such additives in soil and com- Sci. Adv. 3, e1700782.
post. Overall, biodegradable plastics hold promise as a replacement of Gourmelon, G., 2015. Global Plastic Production Rises, Recycling Lags. Worldwatch Insti-
polyethylene mulch, but the release of additives after degradation is of tute, Washington DC http://www.worldwatch.org/global-plastic-production-rises-
recycling-lags-0, Accessed date: August 2018.
concern and demands longer field testing to ensure that either complete Greene, J., 2018. Biodegradation of biodegradable and compostable plastics under indus-
biodegradation occurs or that no long-term harm to the environment is trial compost, marine and anaerobic digestion. Sci. Environ. Bull. 1, 13–18.
caused. Haward, M., 2018. Plastic pollution of the world's seas and oceans as a contemporary
challenge in ocean governance. Nat. Commun. 9. https://doi.org/10.1038/
s41467–018–03104–3.
Author contributions Hayes, D.G., Dharmalingam, S., Wadsworth, L.C., Leonas, K.K., Miles, C., Inglis, D.A., 2012.
Biodegradable agricultural mulches derived from biopolymers. In: Khemani, K.
(Ed.), Degradable Polymers and Materials: Principles and Practice, 2nd edition ACS
H.S., A.B., C.M., M.F. conceived and designed research; H.S., M.F. led Symposium Series. American Chemical Society, Washington, DC, pp. 201–223.
overall study; H.S., M.F. wrote manuscript and analyzed data; H.S., Hayes, D.G., Wadsworth, L.C., Sintim, H.Y., Flury, M., English, M., Schaeffer, S., Saxton, A.M.,
A.B., M.F conducted composting experiments; D.H. contributed FTIR 2017. Effect of diverse weathering conditions on the physicochemical properties of
biodegradable plastic mulches. Polym. Test. 62, 454–467.
analyses; M.E., S.S. contributed δ13C isotope analyses; A.Z., K.S. contrib- Hottle, T.A., Bilec, M.M., Landis, A.E., 2013. Sustainability assessments of bio-based poly-
uted SMPS and miniSPLAT; all co-authors contributed to data interpre- mers. Polym. Degrad. Stab. 98, 1898–1907.
tation and editing. IARC, 2010a. IARC monographs on the evaluation of carcinogenic risks to humans. Carbon
Black, Titanium Dioxide, and Talc. Vol. 93. International Agency for Research on Can-
cer, World Health Organization, Lyon, France.
Acknowledgments IARC, 2010b. Identification of Research Needs to Resolve the Carcinogenicity of High-
priority IARC Carcinogens. International Agency for Research on Cancer, Lyon, France.
ISO, 2012. Specifications for Compostable Plastics. ISO 17088:2012. International Organi-
Financial support for the research was provided by the USDA Spe- zation for Standardization, Geneva, Switzerland.
cialty Crops Research Initiative, CAP (Award 2014-51181-22382), the Kijchavengkul, T., Auras, R., Rubino, M., Ngouajio, M., Fernandez, T., 2008. Assessment of
Western SARE Program (Award 2015-38640-23779), and the NIFA aliphatic-aromatic copolyester biodegradable mulch films. Part I: Field study.
Chemosphere 71, 942–953.
Hatch Projects 1014527 and 1008680. We thank the WSU Franceschi Kijchavengkul, T., Auras, R., Rubino, M., Selke, S., Ngouajio, M., Fernandez, R.T., 2010. Bio-
Microscopy Center for access to their facility. A portion of the research degradation and hydrolysis rate of aliphatic aromatic polyester. Polym. Degrad. Stab.
was performed by A.Z. and K.S. using EMSL (Ringgold ID 130367), a 95, 2641–2647.
Long, C.M., Nascarella, M.A., Valberg, P.A., 2013. Carbon black vs. black carbon and other
DOE Office of Science User Facility sponsored by the Office of Biological
airborne materials containing elemental carbon: physical and chemical distinctions.
and Environmental Research. Bushan Despande of Techmer PM (Clin- Environ. Pollut. 181, 271–286.
ton, TN) provided the carbon black used in PLA/PHA. We thank Michele McGill, E., Oshins, C., 2018. Open source field testing for certified compostable packaging.
BioCycle 59. https://www.biocycle.net/2018/01/11/open-source-field-testing-certi-
Riggs and Cedar Grove Composting, Inc., for allowing us to use the in-
fied-compostable-packaging/.
dustrial composting facility at Maple Valley, WA for the meshbag size Mosnackova, K., Danko, M., Siskova, A., Falco, L.M., Janigova, I., Chmela, S., Vanovcanova,
test. We thank Francesco Degli Innocenti from Novamont, Novara, Z., Omanikova, L., Chodaka, I., Mosnacek, J., 2017. Complex study of the physical prop-
Italy, for contributing the ASTM 5338 test. erties of a poly(lactic acid)/poly(3-hydroxybutyrate) blend and its carbon black com-
posite during various outdoor and laboratory ageing conditions. RSC Adv. 7,
47132–47142.
Appendix A. Supplementary data Müller, Kleeberg, I., Deckwer, W.-D., 2001. Biodegradation of polyesters containing aro-
matic constituents. J. Biotechnol. 86, 87–95.
Prete, M.C., de Oliveira, F.M., Tarley, C.R.T., 2017. Assessment on the performance of nano-
Supporting information includes details on the plastic films, field ex- carbon black as an alternative material for extraction of carbendazim, tebuthiuron,
periment, meshbag preparation, Scanning Mobility Particle Sizer hexazinone, diuron and ametryn. J. Environ. Chem. Eng. 5, 93–102.
(SMPS) and Single Particle Mass Spectrometry (miniSPLAT), stable iso- R Core Team, 2016. R: A Language and Environment for Statistical Computing. R Founda-
tion for Statistical Computing, Vienna, Austria.
tope analysis, effects of meshbags on mulch degradation, and ASTM Rasband, W.S., 2014. ImageJ. U.S. National Institutes of Health, Bethesda, Maryland avail-
D5338 test. Supplementary data to this article can be found online at able online:. http://imagej.nih.gov/ij.
https://doi.org/10.1016/j.scitotenv.2019.04.179. Rudnik, E., Briassoulis, D., 2011. Comparative biodegradation in soil behaviour of
two biodegradable polymers based on renewable resources. J. Polym. Environ.
19, 8–39.
References Saadi, Z., Cesar, G., Bewa, H., Benguigui, L., 2013. Fungal degradation of poly(butyl-
ene adipate-co-terephthalate) in soil and in compost. J. Polym. Environ. 21,
ASTM, 2012. Standard Specification for Labeling of Plastics Designed to be Aerobically 893–901.
Composted in Municipal or Industrial Facilities. ASTM 6400:2012. ASTM Interna- Sintim, H.Y., Flury, M., 2017. Is biodegradable plastic mulch the solution to agriculture's
tional, West Conshohocken, PA. plastic problem? Environ. Sci. Technol. 51, 1068–1069.
ASTM International, 2015. Standard Test Method for Determining Aerobic Biodegra- Skinner, J.J., Beattie, J.H., 1916. A study of the action of carbon black and similar absorbing
dation of Plastic Materials Under Controlled Composting Conditions, Incorporat- materials in soils. Soil Sci. 2, 93–101.
ing Thermophilic Temperatures, D5338–15. ASTM International, West Suyama, T., Tokiwa, Y., Ouichanpagdee, P., Kanagawa, T., Kamagata, Y., 1998. Phylogenetic
Conshohocken, PA. affiliation of soil bacteria that degrade aliphatic polyesters available commercially as
BioBag (2018). Agriculture. Biodegradable Mulch Film. BioBag International AS, https:// biodegradable plastics. Appl. Environ. Microbiol. 64, 5008–5011.
biobagworld.com/products/agriculture, last accessed Oct 2018, Norway. Tokiwa, Y., Calabia, B.P., Ugwu, C.U., Aiba, S., 2009. Biodegradability of plastics. Int. J. Mol.
Bombelli, P., Howe, C.J., Bertocchini, F., 2017. Polyethylene bio-degradation by caterpillars Sci. 10, 3722–3742.
of the wax moth Galleria mellonella. Curr. Biol. 27. https://doi.org/10.1016/j. U.S. Composting Council, 2001. Field Guide to Compost Use. U.S. Composting Council, Be-
cub.2017.02.060. thesda, MD.
 H.Y. Sintim et al. / Science of the Total Environment 675 (2019) 686–693 693

U.S. Department of Health and Human Services, 1988. Occupational Safety and Health Yang, Y., Yang, J., Wu, W.-M., Zhao, J., Song, Y., Gao, L., Yang, R., Jiang, L., 2015. Biodegra-
Guidelines for Carbon Black Potential Human Carcinogen. U.S. Department of Health dation and mineralization of polystyrene by plastic-eating mealworms: part 1. Chem-
and Human Services, Washington, D.C. ical and physical characterization and isotopic tests. Environ. Sci. Technol. 49,
USEPA, 1999. Standards for the use or disposal of sewage sludge. Code of Federal Regula- 12080–12086.
tions Title 40, Part 503. U. S. Environmental Protection Agency, Washington, D.C. Yoshida, S., Hiraga, K., Takehana, T., Taniguchi, I., Yamaji, H., Maeda, Y., Toyohara, K.,
Vroman, I., Tighzert, L., 2009. Biodegradable polymers. Materials 2, 307–344. Miyamoto, K., nd Kohei Oda, Y.K., 2016. A bacterium that degrades and assimilates
Weber, C., Pusch, S., Opatz, T., 2017. Polyethylene bio-degradation by caterpillars? Curr. poly(ethylene terephthalate). Science 351, 1196–1199.
Biol. 27. https://doi.org/10.1016/j.cub.2017.07.004. Zelenyuk, A., Imre, D., Cai, Y., Mueller, K., Han, Y.P., Imrich, P., 2006. SpectraMiner, an in-
Witt, U., Einig, T., Yamamoto, M., Kleeberg, I., Deckwer, W.-D., Müller, R.-J., 2001. Biodeg- teractive data mining and visualization software for single particle mass spectros-
radation of aliphatic-aromatic copolyesters: evaluation of the final biodegradability copy: a laboratory test case. Int. J. Mass Spectrom. 258, 58–73.
and ecotoxicological impact of degradation intermediates. Chemosphere 44, Zumstein, M.T., Schintlmeister, A., Nelson, T.F., Baumgartner, R., Woebken, D., Wagner, M.,
289–299. Kohler, H.-P.E., McNeill, K., Sander, M., 2018. Biodegradation of synthetic polymers in
Worm, B., Lotze, H.K., Jubinville, I., Wilcox, C., Jambeck, J., 2017. Plastic as a persistent ma- soils: tracking carbon into CO2 and microbial biomass. Sci. Adv. 4, eaas9024.
rine pollutant. Annu. Rev. Environ. Resour. 42, 1–26.
Yang, J., Yang, Y., Wu, W.-M., Zhao, J., Jiang, L., 2014. Evidence of polyethylene biodegra-
dation by bacterial strains from the guts of plastic-eating waxworms. Environ. Sci.
Technol. 48, 13776–13784.