Polymer Testing 32 (2013) 60–70

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Polymer Testing
journal homepage: www.elsevier.com/locate/polytest

Material behaviour

Biodegradation behavior of P(3HB,4HB)/PLA blends in real soil

environments
Yun-Xuan Weng a, *, Lei Wang a, Min Zhang a, Xiu-Li Wang b, Yu-Zhong Wang b
a
Beijing Technology & Business University, Beijing 100037, China
b
Center for Degradable and Flame-Retardant Polymeric Materials, College of Chemistry, Sichuan University, Chengdu 610064, China

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

Article history: Both poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB,4HB)] and poly(lactic acid)

Received 19 August 2012 (PLA) are biobased polymers, and their blends can be used to prepare biodegradable
Accepted 28 September 2012 polymeric materials with good comprehensive properties. In this paper, the biodegrada-
tion behavior of P(3HB,4HB)/PLA blends in real soil environments was for the first time
Keywords: investigated via appearance analysis, SEM, FTIR and elemental analysis of their degraded
P(3HB,4HB)
residues. It was found that P(3HB,4HB) and P(3HB,4HB)/PLA blends can be biodegraded
PLA
under real soil conditions. The order of biodegradability is as follows: PHA-100
Soil condition
Degradation [P(3HB,4HB)] > PHA-75 [P(3HB,4HB)/PLA, 75/25] > PHA-50 [P(3HB,4HB)/PLA, 50/
50] > PHA-25 [P(3HB,4HB)/PLA, 25/75] > PLA, which correlates well with the PLA content.
Due to the different degradation mechanisms, P(3HB,4HB) and PLA have the fastest
degradation rate in 20 and 40 cm soil depth, respectively. Their blends also have different
degradation rates in different depths of soil.
Ó 2012 Elsevier Ltd. All rights reserved.

1. Introduction Moreover, the biodegradation rate of the blends in soil was

Polyhydroxyalkanotes (PHAs) are a family of biobased
and biodegradable polymers [1–8], in which poly(3-
hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB,4HB)] has
been commercialized by Tianjin GreenBio Materials Co., Ltd
(GreenBio, China) with a scale of 10,000 tones per year. By
changing the molar ratio of 3HB to 4HB, P(3HB,4HB) can be
adjusted from highly crystalline plastics to soft elastomers
[9]. In order to widen its application field, it can be blended
with other polymers such as poly(lactic acid) (PLA) [10–14].
Wang et al. prepared PLA/PHBV blends, and found that the
toughness and elongation at break of the PLA/PHBV (70/30)
blends were greatly improved by the addition of PEG.
* Corresponding author. Beijing Technology & Business University, 1001
Room, 11th Building, 11 Fucheng Road, Beijing 100037, China. Tel.: þ86 10
68985380; fax: þ86 10 68983573.
E-mail addresses: wyxuan@th.btbu.edu.cn, wengyx@degradable.org.
cn (Y.-X. Weng).
also accelerated due to the introduction of PEG [15].
The biodegradation behavior of PHAs, PLA and their
blends has been investigated by some researchers. Weng
et al. [16] investigated the biodegradation behavior of PHAs
with different chemical structures under controlled com-
posting conditions. It was found that the degree of biodeg-
radation of P(3HB, 4HB) was 90.3% according to ISO 14855-1.
Rudnik and Briassoulis [17,18] compared the biodegradation
of PLA and PHA in soil environment and in composting
conditions. The results showed that PHA polymer could be
biodegraded in both real soil environment and composting
conditions with a high degradation rate. However, the
degradation rate of PLA in soil was slow even if it was
biodegradable under composting conditions. Kale et al. [19]
investigated the degradation of PLA under real composting
(65
C) and simulated (58
C) conditions. They found that
PLA was biodegradable but needed longer time and/or
higher temperature (at least 58
C) to degrade. The
biodegradation of PHBV/PP blends in soil showed that PHBV
was biodegradable while PP could not be biodegraded [20].

0142-9418/$ – see front matter Ó 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.polymertesting.2012.09.014
 Y.-X. Weng et al. / Polymer Testing 32 (2013) 60–70 61

Although there has been some literature focusing on the

biodegradation of PHA, PLA, or their blends in composting
conditions, the biodegradation behavior of P(3HB,4HB)/PLA
blends in real soil environments has not been reported. In
this paper, we will report for the first time the biodegrad-
ability of P(3HB,4HB)/PLA blends with different P(3HB,4HB)
contents in real soil environments.

2. Experimental

2.1. Materials

Fig. 2. Curve of temperature variation of soil with time.

P(3HB,4HB) powder (containing 10% mol 4HB) was
supplied by Tianjin Greenbio Material Co.(Tianjin, China). The
material has a number-average molecular weight (Mn) of
44.6  104 g/mol, as determined by gel permeation chroma-
tography (GPC), its glass transition temperature (Tg) is 3
C and
its melting point (Tm) is 164
C, as determined by differential
scanning calorimetry (DSC). PLA, NatureWorksÒ PLA (2002D),
has a weight-average molecular weight (Mn) of 11.0  104 g/
mol, and its Tg and Tm are 60
C and 156
C, respectively.
2.2. Preparation of sample films
PLA pellets and P(3HB,4HB) powders were premixed in
a high-speed mixer at 400 rpm for 5 min, and the mixtures
were subsequently melt-extruded via a co-rotating twin-
screw extruder (PIE-35, Coperion Keya Machinery Co. Ltd.,
Nanjing, China) with a L/D ratio of 40, operated at 160
C
with a screw speed of 90 rpm. Bends with different weight
ratios of P(3HB,4HB)/PLA, 100/0, 75/25, 50/50, 25/75 and 0/
100 were produced, and coded as PHA-100, PHA-75, PHA-
50, PHA-25 and PLA, respectively. The Films were
prepared from the blends by extrusion casting with a single
screw extruder (LP-S-50, LabTech, Sweden).
Films were cut into 5 cm  10 cm specimens for testing
under real soil conditions.

Fig. 1. The schematic diagram and photos of the soil pond: (a) Schematic diagram (b) digital photo before testing (c) digital photo after testing.
62 Y.-X. Weng et al. / Polymer Testing 32 (2013) 60–70

2.3. Biodegradation under soil conditions Beijing, China; Fig. 1 shows a schematic diagram and digital
photos.
Biodegradation under real soil conditions was carried The actual size of the soil pond is 4.5 m  2.5 m  1.5 m
out at the experimental soil pond located in the west of (length  width  height). At the bottom of the pond wall

Fig. 3. The apparent variation of P(3HB,4HB)/PLA films during the burial in 20 cm depth soil: (a) PHA-100 (b) PHA-75 (c) PHA-50 (d) PHA-25 (e) PLA.
 Y.-X. Weng et al. / Polymer Testing 32 (2013) 60–70 63

there are three weep holes in the two opposite sides
through which excess water can ooze out, and air can get in
and out. Field soil was put into the pond until the surface
was 20 cm lower than the top of the pond wall. Samples
with different compositions were buried in the soil, and
located at depths of 20 cm and 40 cm. Five specimens were
tested for each blend film.
After the samples were buried, tall fescue seeds were
planted on the soil surface. The plants were watered
regularly so as to imitate the real soil in the natural

Fig. 4. The apparent variation of P(3HB,4HB)/PLA films during the burial in 40 cm depth soil: (a) PHA-100 (b) PHA-75 (c) PHA-50 (d) PHA-25 (e) PLA.
64 Y.-X. Weng et al. / Polymer Testing 32 (2013) 60–70

environment. The soil temperature at 20 cm depth was 3. Results and discussion

recorded every day. One specimen of each sample was
taken out every month for testing. 3.1. Temperature of soil in 20 cm depth

2.4. Characterization The soil temperature at 20 cm depth was recorded

The surface morphology of the samples biodegraded for
different times was recorded by a digital camera (Sony DSC-
W80, Japan) and scanning electron microscope operated at
3.0 kV (TESCAN VEGA II, TESCAN, s.r.o, Czech). The surfaces
of all the samples were coated with gold prior to SEM
examination. FT-IR spectra of the samples were obtained
with an Avater 370 FT-IR spectrophotometer (Nicolet Co.,
USA) using a potassium bromide pellet. The C, H, O contents
of samples were determined with an elemental analyzer
(vario EL cube, Germany).
everyday until the test was terminated.
From Fig. 2, it can be seen that the temperature of the
soil was mainly at 20  3
C, which was suitable for the
living and reproduction of bacteria.
3.2. Appearance changes of P(3HB,4HB)/PLA films before and
after soil burial
The changes in appearance of P(3HB,4HB)/PLA films
during the burial in 20 and 40 cm of soil were observed by
digital camera, and the photos are shown in Figs. 3 and 4.

Fig. 5. SEM pictures of P(3HB,4HB)/PLA films at different degradation time buried in 20 cm depth: (a) PHA-100 (b) PHA-75 (c) PHA-50 (d) PHA-25 (e) PLA.
 Y.-X. Weng et al. / Polymer Testing 32 (2013) 60–70
It can be seen from Fig. 3 that the size of all P(3HB,4HB)/
PLA films with different PLA contents become smaller with
the increase of burial time at 20 cm depth of soil. After 1
month, PHA-100 and PHA-75 films lost their integrated
appearance, and PHA-100 film was difficult to collect,
indicating that its degradation rate is faster than those of
the others. After 2 months, the PHA-100, PHA-75 films
were almost biodegraded and could not be collected, while
PHA-50 became fragmented. After 3 months, the residue of
PHA-50 film was very small, and PHA-25 and PLA films
began to break into small fragments. After 5 months, only
PLA fragments were found in the soil. During testing, the
films were either digested by the microorganisms or
broken into small fragments. Higher material loss and
greater appearance change reflected the higher biodegra-
dation rate. Based on the appearance variations in Fig. 3,
the order of biodegradation rate can be preliminarily
determined as follows: PHA-100 > PHA-75 > PHA-
50 > PHA-25 > PLA.
When the samples were buried at the 40 cm depth, their
size also became smaller with the increase of testing time.
After 1 month, PHA-100 film lost its integrated appearance.
After 2 months, PHA-100, PHA-75 films were biodegraded
to small fragments. PHA-50, PHA-25 and PLA began to be
disintegrated. After 3 months, it was difficult to find the
residual PHA-100, while PHA-75 and PHA-50 became
Fig. 6. SEM pictures of P(3HB,4HB)/PLA films at different degradation time buried in 40 cm depth: (a) PHA-100 (b) PHA-75 (c) PHA-50 (d) PHA-25 (e) PLA.
65
smaller fragments. After 5 months, only PHA-25, PLA
fragments could be found in the soil. Based on the results,
the order of biodegradation rate can be summarised as
follows: PHA-100 > PHA-75 > PHA-50 > PHA-25 > PLA,
which was the same as that for 20 cm depth soil.
Compared the results shown in Figs. 3 and 4, it was
found that the biodegradation rates of PHA-100, and PHA-
75 buried at 20 cm depth were higher than those at 40 cm.
However, the biodegradation rates at 40 cm of soil of PHA-
50, PHA-25 and PLA were faster, illustrating that when the
PLA content was increased the blends were more easily
degraded in 40 cm of soil, which can be ascribed to the
anaerobic conditions at 40 cm depth of soil which were
more favorable for the biodegradation of PLA.
3.3. Micro-morphology of P(3HB,4HB)/PLA films before and
after soil burial
The micro-morphologies of P(3HB,4HB)/PLA films
during degradation at 20 and 40 cm of soil were investi-
gated by SEM (shown in Figs. 5 and 6).
It can be seen from Fig. 5 that the surfaces of
P(3HB,4HB)/PLA films were smooth before degradation.
After being buried in 20 cm of soil for 1 month, the surface
of PHA-100, PHA-75 and PHA-50 were eroded and became
rough. The surfaces of PHA-100 were seriously eroded after
66 Y.-X. Weng et al. / Polymer Testing 32 (2013) 60–70

2-month burial. In fact, after 2 months it was difficult to
find the PHA-100 fragments, and this phenomenon can
only be observed for PHA-75 after 3-month burial. After 3
months burial, the surface of PHA-50 film was obviously
eroded and the surface of PHA-25 also became rough. The
surface of PLA film began to be eroded after 4 months. After
4-month burial, many small gullies appeared on the
surfaces of PHA-25 and PLA films. From Fig. 5, we can see
clearly that the appearance of degraded P(3HB,4HB)/PLA
surfaces was different from that of PLA. The surface of
degraded PHA-100 film showed many large cavities after 1
month burial. In addition, with the increase of PLA content,
the roughness of the degraded blend surface lessened,
indicating that the degradation rate was decreased. For
pure PLA film, the serious surface corrosion cannot be
observed even after 4 months. This phenomenon illus-
trated that the degradation of P(3HB,4HB) was firstly
caused erosion by microorganisms starting from the
surface, and then spreading gradually to the interior. PLA
is more easily degraded by hydrolysis rather than enzy-
matic degradation, and its degradation products can be
totally digested by microorganisms such as fungi or
bacteria [21–25]. The order of biodegradation rate deter-
mined from the SEM was as same as that recorded by the

Fig. 7. The FT-IR spectra of P(3HB,4HB)/PLA films before and after degradation: (a) PHA-100; (b) PHA-75; (c) PHA-50; (d) PHA-25; (e)PLA.
 Y.-X. Weng et al. / Polymer Testing 32 (2013) 60–70
digital photos. When the samples were buried in 40 cm of
soil, their micro-morphology variations were similar to
those in 20 cm of soil.
3.4. FT-IR spectra of P(3HB,4HB)/PLA films before and after
burial
The chemical structure changes of P(3HB,4HB)/PLA
films before and after degradation were investigated by FT-
IR (shown in Fig. 7).
It can be seen from Fig. 7 that P(3HB,4HB)/PLA films
before and after burial showed similar absorption peaks in
their FTIR spectra. For PHA (Fig. 7a), before burial, the
characteristic peak at 1735 cm1 was assigned to C]O
stretching vibration, the peaks at 1456 cm1 and 1381 cm1
were assigned to CH3 deformation vibration and C–O–C
stretching vibration, and the peaks at 1089 cm1 and
1057 cm1 were ascribed to the formation of the 21-helix
structure of PHAs [26]. However, after it was buried in
20 cm or 40 cm of soil, the absorption peak of C]O
stretching vibration was shifted from 1735 cm1 to
Fig. 7. (continued)
67
1759 cm1. The other characteristic peaks were nearly
unchanged. Similar FT-IR spectra were obtained for
P(3HB,4HB)/PLA films with different PLA contents.
For PLA (Fig. 7e), before burial, the absorption peak of
C¼O stretching vibration was found at 1749 cm1, –CH–
deformation vibration was observed at 1457 cm1 and
1386 cm1, and –C–O– stretching vibration was found at
1186 cm1. After 4-month burial in 20 cm of soil, the
absorption peak of C]O stretching vibration was shifted to
1758 cm1. However, the absorption peak of C]O
stretching vibration in 40 cm was almost unchanged. These
phenomena demonstrated that the degradation mecha-
nism of PLA buried in different depths of soil is different.
The degradation of PLA in 20 cm of soil was mainly aerobic
biodegradation, and that in 40 cm of soil was anaerobic
biodegradation.
For all samples, it was found that the peaks in the 4000-
3000 cm1 region became broadened after degradation,
which was due to the formation of hydroxyl and carboxylic
groups. This was also confirmed by the shift of the absorp-
tion peak of C]O stretching vibration after degradation.
68 Y.-X. Weng et al. / Polymer Testing 32 (2013) 60–70
The FTIR results demonstrated that the biodegradation
of P(3HB,4HB) films was mainly caused by microorganisms
and many low molecular weight polymers with carboxyl
groups were produced [27–30]. As far as PLA are concerned,
its different degradation mechanisms produced different
degraded products. Lower molecular weight polymers with
carboxyl groups were produced during aerobic biodegra-
dation. However, during anaerobic biodegradation, high
molecular weight PLA’s were degraded into lower molec-
ular weight ones, which made C]O stretching vibration
unchanged.
3.5. Element content of P(3HB,4HB)/PLA films before and after
burial
The element contents of C, H, O of P(3HB,4HB)/PLA films
before and after burial were determined, and the results
were listed in Table 1.
Fig. 7. (continued)
From Table 1, it can be seen that the carbon content of all
samples decreased, while their oxygen content increased
after degradation. When PHA-100 and PHA-75 were
degraded in 20 cm of soil for 1 month, their carbon contents
were decreased from 55.8% to 32.9%, 53.7% to 48.8%,
respectively. When they were degraded in 40 cm of soil,
however, their carbon contents were decreased to 46.1% and
49.7%, respectively. These results demonstrate that both
PHA-100 and PHA-75 can degrade in soil conditions and the
degradation rate of PHA-100 is higher than that of PHA-75.
Furthermore, the degradation rates of PHA-100 and PHA-
75 in 20 cm of soil are higher than those at 40 cm depth.
The decrease of carbon content for PHA-50 could be
observed after 3 month burial, its value decreased from 52.2%
to 42.4% or 44.6% in 20 or 40 cm of soil. After 3 month burial in
40 cm of soil, the carbon content for PLA was decreased from
50.0% to 47.5%. However, when it was degraded in 20 cm of
soil, its carbon content was little decreased (from 50.0% to
 Y.-X. Weng et al. / Polymer Testing 32 (2013) 60–70 69

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