Colloids and Surfaces B: Biointerfaces 144 (2016) 38–45

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Colloids and Surfaces B: Biointerfaces

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

Construction of a controlled-release delivery system for pesticides

using biodegradable PLA-based microcapsules
Baoxia Liu a,1 , Yan Wang b,1 , Fei Yang a , Xing Wang a , Hong Shen a , Haixin Cui b,∗ ,
Decheng Wu a,∗
a
Beijing National Laboratory for Molecular Sciences, State Key Laboratory of Polymer Physics & Chemistry, Institute of Chemistry, Chinese Academy of
Sciences, Beijing 100190, China
b
Institute of Environment and Sustainable Development in Agriculture, Chinese Academic of Agriculture Sciences, Beijing 100081, China

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

Article history: Conventional pesticides usually need to be used in more than recommended dosages due to their loss
Received 18 November 2015 and degradation, which results in a large waste of resources and serious environmental pollution. Encap-
Received in revised form 29 March 2016 sulation of pesticides in biodegradable carriers is a feasible approach to develop environment-friendly
Accepted 31 March 2016
and efficient controlled-release delivery system. In this work, we fabricated three kinds of polylactic acid
Available online 1 April 2016
(PLA) carriers including microspheres, microcapsules, and porous microcapsules for controlled delivery of
Lambda–Cyhalothrin (LC) via premix membrane emulsification (PME). The microcapsule delivery system
Keywords:
had better water dispersion than the other two systems. Various microcapsules with a high LC contents
Controlled release
Water dispersion
as much as 40% and tunable sizes from 0.68 to 4.6 ␮m were constructed by manipulating the process
Biodegradable parameters. Compared with LC technical and commercial microcapsule formulation, the microcapsule
Microcapsule systems showed a significantly sustained release of LC for a longer period. The LC release triggered by LC
Thermal-stability diffusion and matrix degradation could be optimally regulated by tuning LC contents and particle sizes of
UV-shielding properties the microcapsules. This multi-regulated release capability is of great significance to achieve the precisely
controlled release of pesticides. A preliminary bioassay against plutella xylostella revealed that 0.68 ␮m
LC-loaded microcapsules with good UV and thermal stability exhibited an activity similar to a commercial
microcapsule formulation. These results demonstrated such an aqueous microcapsule delivery system
had a great potential to be further explored for developing an effective and environmentally friendly
pesticide-release formulation.
© 2016 Elsevier B.V. All rights reserved.

1. Introduction Conventional formulations usually rapidly fall below the effec-

Currently, about 2 million tons of pesticides are applied to con-
trol pathogens and pests every year all over the world. However, it
is estimated that ∼90% of the pesticides actually are lost due to their
degradation, photolysis, evaporation and surface runoff, and only
∼0.1% of the pesticides are finally deposited on the harmfully bio-
logical targets [1,2]. This long-term extensive and inefficient use of
pesticides has caused serious social concerns on food safety and
ecological environment [3–6]. The social concerns motivate the
researchers to develop efficient, safe and green pesticide formu-
lations [7–9].
∗ Corresponding authors at: No.2, Zhongguancun North First Street Haidian Dis-
trict, Beijing 100190, China.
E-mail addresses: cuihaixin@caas.cn (H. Cui), dcwu@iccas.ac.cn (D. Wu).
1
These authors contributed equally to this work.
tive level after an initial burst release of pesticide. A controlled
delivery system could achieve continuous and stable release of
active ingredients, maintaining a predetermined minimum effec-
tive level of drugs for a specified period of time [10–12], so it is
an effective way to improve the utilization of pesticide via pro-
longing the effective duration with reduced spraying times and
pesticide dosages [13,14]. Controlled-release technology of pes-
ticides is gradually developing from the simple and qualitative
release toward the precise and quantitative controlled release,
ultimately achieving the more economic, safe and effective insect
control and reducing environmental pollution.
Encapsulation of pesticides into polymeric carriers has attracted
emerging interests. The polymeric matrix prevents direct expo-
sure of pesticides to environment, reducing loss of evaporation and
degradation. Biodegradable polymers, including chitosan [15,16],
alginate [8,9], polyacrylamide and starch [17,18] et al., has been
widely investigated for constructing pesticide carriers [19]. Polylac-

http://dx.doi.org/10.1016/j.colsurfb.2016.03.084
0927-7765/© 2016 Elsevier B.V. All rights reserved.
 B. Liu et al. / Colloids and Surfaces B: Biointerfaces 144 (2016) 38–45
Fig. 1. Schematic description for preparation of microspheres, microcapsules and porous microcapsules (A) and PME process (B).
tic acid (PLA) is a FDA-approved material widely used as drug/cell
carriers in the medical field [20–26]. However, PLA has been rarely
reported and studied systemically as carrier materials in the field
of pesticide. The final metabolized products of PLA in vivo are car-
bon dioxide and water, which have no harm to human and the
environment. Industrial PLA is cheap (∼US$3/kg) and affordable as
pesticide carriers. The molecular weights, physical properties and
degradation rates of PLA can be tuned to optimize release proper-
ties of pesticides [27–30]. It is preferred to construct PLA carriers
for development of safe and green pesticide formulations.
The conventional encapsulation techniques of pesticides
include in situ polymerization [31], interfacial polycondensation
[32,33], nanoprecipitation [34], suspension cross-linking [35], and
solvent evaporation/extraction [36]. However, these techniques
usually require complicated process and rigorous conditions. The
size of particles is difficult to control and the size distribution is
very broad via mechanical stirring, homogenization or ultrasoni-
cation. Poor dispersion and uniformity as well as residual organic
solvents would hinder the applications of the resulting delivery sys-
tems. Good dispersion and uniform size of the carriers can improve
their adhesion on the surface of foliage and permeability to harm-
ful insects, increasing the utilization rate and biological activity of
pesticides [37–39]. Premix membrane emulsification (PME) tech-
nology is a novel and simple method to prepare uniform particles
[19,40–42]. The whole procedure of PME involves using an applied
pressure to force coarse emulsion to pass through the membrance
without intense ultrasound and heat, which avoids the degradation
of pesticides under turbulent ultrasound and high temperature.
The resulting particle size in a range from hundreds nanometers to
micrometers is primarily controlled by membrane pore size as well
as the process parameters including applied membrane, membrane
pressure and viscosity of emulsion, which is suitable to obtain var-
ious reproduced carries [43,44]. In a word, the PME is a promising
industrialized technique in construction of pesticide-delivery sys-
tems because of its distinguishing characters such as low energy
consumption, easy operation and simple equipment, amendable
for large scale production [45].
The reported microcapsules and commercial microcapsules
for pesticides are generally in micron scale and inhomogeneous.
Herein, we adopt the PME combining with the emulsion method
39
to fabricate uniform PLA-based carriers with tunable sizes from
0.68 to 4.6 ␮m for controlled release of pesticides. Several deliv-
ery systems, employing microcapsules, microspheres, and porous
microcapsules, were fabricated and investigated. The results
showed that the microcapsule-based delivery system had better
dispersion, high loading content and entrapment efficiency, an
effective size-controlled property, and good ultraviolet (UV) shield-
ing and stability.
2. Materials and methods
2.1. Materials
Polylactide (PLA) and Lambda–Cyhalothrin (LC) were kindly
provided by Dongguan Zhuyou Plastic Co., Ltd. (Dongguan, China),
and Yangnong Chemical Co., Ltd. (Yangzhou, China), respectively.
Bovine serum albumin (BSA) was obtained from Beijing Biodee
Biotechnology Co., Ltd. (Beijing, China). Poly(vinyl alcohol) (PVA)
with a molecular weight of 30,000–70,000 and a hydrolysis of
87–89% was purchased from Sigma-Aldrich (St. Louis, MO). The
specification of the dialysis membrane purchased from Beijing
Tianan Technology Co., Ltd., (Beijing, China) was 50,000. Other
chemical reagents were of analytical grade and purchased from
Beijing Chemical Works (Beijing, China).
2.2. Preparation of the LC-loaded microcapsules
The PLA microcapsules were prepared via a water-in-oil-in-
water (W1 /O/W2 ) double-emulsion method combined with PME.
Briefly, PLA and LC were dissolved in methylene chloride as the oil
phase. The oil phase was mixed with deionized water and sonicated
to form the primary emulsion of W1 /O. Then, the primary emulsion
was immediately poured into a large volume of aqueous PVA solu-
tion under mechanical stirring to prepare coarse double emulsion.
The coarse double emulsion was extruded through an SPG mem-
brane under a certain nitrogen pressure several times. The uniform
double emulsion was obtained and solidified under magnetic stir-
ring overnight. The hardened microcapsules were collected via
centrifugation and washed with deionized water three times. The
obtained microcapsules were freeze-dried using a lyophilizer to
40 B. Liu et al. / Colloids and Surfaces B: Biointerfaces 144 (2016) 38–45
Fig. 2. SEM images of the (A) microspheres, (B) microcapsules, and (C) porous
microspheres before and after cut by a super thin blade and optical photos of their
suspensions (D) in water after setting for 4 h. The concentrations both are 2 mg/mL,
respectively.
yield a free-flowing powder. The dried powder was stored at 4 ◦ C
prior to use.
The microcapsules with different sizes were prepared with var-
ious optimal process parameters. The 4.6 ␮m microcapsules were
obtained with 100 mg/mL PLA in the oil phase, 1 wt.% PVA concen-
tration in the external water phase, and 4 circulations through a
9.0 ␮m SPG membrane under 80 KPa pressure. The 2.4 ␮m micro-
capsules were produced with 50 mg/mL PLA in the oil phase, 1 wt.%
PVA concentration in the external water phase, and 4 circulations
through the 7.2 ␮m SPG membrane under 80 KPa pressure. The
0.68 ␮m microcapsules were yielded with 30 mg/mL PLA in the oil
phase, 0.1 wt.% PVA concentration in the external water phase, and
3 circulations through the 1.0 ␮m SPG membrane under 1000 KPa
pressure. The feed weight ratios of LC/PLA were 1/9, 1/4, 3/7, 2/3 and
1/1 to produce various microcapsules with different LC contents for
further study.
The preparation process of LC-loaded porous microcapsules
(50 wt.% LC) was the same as that of producing 4.6 ␮m microcapsule
except that 0.4 g BSA was added into 4 mL of the inner water phase
to induce formation of micropores on surface of the microcapsules.
2.3. Preparation of the microspheres
The PLA microspheres were prepared via an oil-in-water (O/W)
emulsion method combined with PME. Briefly, PLA and LC were
dissolved in methylene chloride. The solution was immediately
poured into an aqueous PVA solution under mechanical stirring to
prepare the coarse double emulsion. The coarse double emulsion
was then extruded through an SPG membrane under a certain nitro-
gen pressure several times. The uniform emulsion was obtained
and solidified under magnetic stirring overnight. The hardened
microspheres were collected via centrifugation and washed with
deionized water three times. The microspheres were further freeze-
dried to yield a free-flowing powder. The dried powder was stored
at 4 ◦ C prior to use. The 4.6 ␮m LC-loaded microspheres (50 wt.%
LC) were prepared by the following optimal process parameters:
200 mg/mL PLA in the oil phase, 1 wt.% PVA concentration in the
water phase, and 4 circulations through a 9.0 ␮m SPG membrane
under 100 KPa pressure.
2.4. Characterization of the LC-loaded microcapsules and
microspheres
The morphology of the microparticles was observed via scan-
ning electron microscopy (SEM, JSM-6700F; JEOL Ltd., Tokyo, Japan)
with an accelerating voltage of 5 kV and a working distance of 8 mm.
The sample was mounted to metal stubs using double-sided tape
and vacuum-coated with a thin layer of platinum using a sputter
coater (EM SCD 500; Leica, GER). The sizes of the microparticles
were measured with laser scatter using a zetasizer (Zetasizer Nano
ZS90; Malvern Instruments Ltd., Malvern, UK).
2.5. Drug content and entrapment efficiency
The loading content and entrapment efficiency of LC in the
formulations were determined by high-performance liquid chro-
matography (HPLC) (Agilent 1260, Agilent Technologies, Santa
Clara, CA). An approximately 30 mg sample was fully dissolved
in 30 mL of methylene chloride. The solution was added to ethyl
alcohol and dried via reduced-pressure distillation. Then, 5 mL
of HPLC-grade ethanol was added to soak the LC from the dried
precipitation. Finally, the mixture was filtered to form a clear
solution for HPLC analysis. The mobile phase consisting of ace-
tonitrile/water (90:10) was applied. A reverse-phase Inertsil C-18
column (150 mm × 4.6 mm, pore size 5 mm, GL Science Inc., Tokyo,
Japan) was used. The flow rate of the mobile phase was set at
1 mL/min. The sample was detected at 230 nm with a UV–vis detec-
tor.
2.6. In vitro release of LC
To study in vitro release of the encapsulated LC from the micro-
capsules, a 10 mL of LC suspension in an ethanol/water mixture
 B. Liu et al. / Colloids and Surfaces B: Biointerfaces 144 (2016) 38–45 41

Fig. 3. SEM images of LC-loaded microcapsules (A) MC1, (B) MC2, and (C) MC3 with average sizes of 4.6, 2.4 and 0.68 ␮m and PDIs of 0.122, 0.090 and 0.066, respectively.

Table 1 where Y is the mortality rate of treated group. X is the logarithm of

The LC loading content and loading efficiency of the microcapsules with different
pesticide dosage. LC50 means the dosage when the mortality rate
feed ratios of PLA/LC.
of the treated group was 50%.
Weight contents of LC (%) Loading content (%) Loading efficiency (%)

10 9.5 95.0 2.8. Stability test

20 18.2 89.3
30 28.7 86.2
40 34.4 83.6 The microcapsules were packed in glass tubes and stored at
50 41.0 82.0 4 ◦ C for 7 days and 50 ◦ C for 14 days, and the changes in load-
ing contents and surface of the microcapsules were studied. The
UV-shielding properties of the microcapsules were tested as fol-
Table 2 lows. A certain amount of LC-loaded microcapsules were mixed
Constants from fitting the generalized model of Eq. (1) to the release data of LC from
with an ethanol/water mixture (70:30, v/v), and the suspension
the different capsule suspensions.
was transferred into a culture dish. In the center of the reactor, a
Sample r n T50 500 W (Emax = 365 nm) UV lamp was applied to the sample for a
MC1 0.9891 0.73 16.0 desired period of time. The temperature was maintained at 25 ◦ C
MC2 0.9914 0.82 14.5 during the experiments. At various time intervals, the culture dish
MC3 0.9942 0.83 14.1 was removed from the reactor, and the content of LC in the culture
MC4 0.9930 0.72 18.6
dish was determined using the method described above. The con-
MC5 0.9934 0.75 16.2
MC6 0.9954 0.65 14.6 centration of LC was measured with a UV–vis spectrophotometer
at a wavelength of 280 nm to determine the kinetic profile of the
released LC.

(30:70, v/v) was loaded in the dialysis membrane. Then the mem-
brane was placed into a wild-mouth flask with 90 mL of the mixed
solution of ethanol and water. The flask was incubated in the
incubator shakers under shaking at 300 rpm. Next, 2 mL of the
mixed solvent outside the dialysis membrane was withdrawn over
a period of 250 h at various time intervals and was replaced with a
fresh mixed solution. The concentration of LC was measured using a
UV–vis spectrophotometer at a wavelength of 280 nm to determine
the kinetic profile of the released LC.
2.7. Biological assay
A preliminary bioassay of LC-loaded microcapsules with various
sizes (from 0.68 to 4.6 ␮m) against plutella xylostella was evaluated
using a commercial formulation as control. The LC-loaded micro-
capsules and commercial formulation with a LC concentration to be
25 g/L were diluted into different concentrations with 0.1% triton
to prepare the test suspensions according to statistical require-
ments. The 2rd in stars of plutella xylostella were immersed in the
prepared test suspensions for 10 s, and then were placed on cab-
bage leaves in petri dishes to investigate the mortalities after 48 h.
Assessments were made on a dead/alive basis, and mortality rates
were corrected using Abbott’s formula. SPSS software was used to
calculate toxicity regression equations, LC50 and confidence limits.
The toxicity regression equation was given as below:
Y = bX + a
3. Results and discussion
3.1. Preparation of the LC delivery systems
The structure of the delivery system is one of the most key
factors affecting the dispersion of pesticide. Good uniformity and
dispersion of the pesticide-delivery system is conducive to improv-
ing adhesion and permeability of the pesticide on target crops
and to achieving effective utilization and high bioavailability of
the pesticide. To obtain an optimal delivery system with stable
and homogeneous dispersion, we constructed three kinds of LC-
delivery systems based on microspheres, microcapsules and porous
microcapsules, by combining the emulsion method (O/W for micro-
spheres and W/O/W for microcapsules) with PME and the osmosis
induction method for formation of the porous structures, as illus-
trated in Fig. 1.
Fig. 2 depicts the morphologies and cross-sections of the var-
ious delivery systems. The microspheres (Fig. 2A) had an almost
identical spherical shapes and smooth surfaces as the microcap-
sule systems (Fig. 2B), but a distinct solid inner structure instead of
a hollow inner cavity for the microcapsule system from the cross-
sectional images. The results demonstrated the O/W emulsion
and W1 /O/W2 double-emulsion methods combined with PME can
successfully prepare solid-microsphere and hollow-microcapsule
delivery systems, respectively. During the preparation of the
microcapsule-based delivery systems, addition of porogen BSA in
the inner water phase was favorable to form porous microcapsules.
Fig. 2C indicated that some small pores appeared in the surfaces
42 B. Liu et al. / Colloids and Surfaces B: Biointerfaces 144 (2016) 38–45

Fig. 4. Effects of the sizes (A); 4.6, 2.4 and 0.68 ␮m for MC1, MC2 and MC3 with 50% feeding contents of LC) and LC loading contents (B); 22%, 28% and 45% for MC4, MC5
and MC6 constructed using 1 ␮m SPG membrane) on the release behavior of LC, and SEM images of surface morphology of the MC1 before (C) and after (D) soaking in an
ethanol/water mixture (30:70, v/v) for 250 h.

of the microcapsules. The formation of the pores should result
from water shift under osmotic pressures between the internal and
external water phases after introduction of BSA in the inner water
phase. Some water molecules go into the inner, also resulting in
formation of bigger hollow cavity than the microcapsules. It must
note that the entire PME procedure used an applied pressure to
force coarse emulsion to pass through a membrane without intense
ultrasound or heat, avoiding the degradation of the pesticide under
turbulent-ultrasound and high-temperature conditions.
The dispersion capacity and stability of the pesticide deliv-
ery systems in water are vital for successful construction of their
aqueous formulations. The aqueous microcapsule suspension could
maintain stable and homogeneous even after standing for 4 h
instead of obvious deposition observed from the microsphere and
porous microcapsule suspensions, resulting from different gravity
effects as shown in Fig. 2D. The microcapsule is light and has high
buoyancy in water due to its hollow structure, which is favorable to
slow down the settling of the microcapsule. The microsphere with
a solid structure possesses high density, resulting in its rapid set-
tlement. For the porous microcapsule suspension, the open pores
allow water to go into the inner cores, causing a loss of buoyancy
and subsequently accelerated precipitation. So the microcapsule is
optimal to be further studied for developing the aqueous formula-
tion of pesticides.
 B. Liu et al. / Colloids and Surfaces B: Biointerfaces 144 (2016) 38–45
Fig. 5. The LC50 values of the MC1, MC2, MC3 and commercial formulations in
bioassay study.
3.2. The loading content and entrapment efficiency of LC
The loading content and entrapment efficiency of pesticides in
the microcapsules have an important effect on overall performance
of the pesticide delivery system. High entrapment efficiency and
loading content are preferred to reduce the waste of pesticide dur-
ing the preparation process as well as to avoid its extensive use
in spraying and subsequent environmental pollution. To achieve
high loading content as well as suitable entrapment efficiency, we
constructed several specimens by feeding various weight ratios of
PLA/LC. Table 1 indicated that all the specimens had loading effi-
ciencies of higher than 82%. The results demonstrate the mild PME
process can effectively avoid damage to the pesticide activity with-
out a prolonged period of intense ultrasound and/or heating. As the
feeding weight contents of LC increased from 10% to 50%, the load-
ing contents of LC increased from 9.5% to 41.0% with the loading
Fig. 6. Comparison of the LC photolysis percentage of LC technical and MC3 formulations under UV irradiation (A) and LC contents (B) and its SEM images of the MC3
microcapsules before (C), and after 4 ◦ C for 7 days (D) and 54 ◦ C for 14 days (E).
43
efficiency decreased from 95% to 82%. It is reasonable as the LC feed-
ing contents increased, the corresponding reduced concentrations
of PLA lowered the LC encapsulation ability, subsequently lead-
ing to decrease of the loading efficiency. However, even for 50 wt%
feeding contents of LC, the loading efficiency still can reach as high
as 82%. So, we chose 50 wt% feeding contents of LC to produce the
LC delivery systems with high loading contents.
3.3. Preparation of the LC-loaded microcapsules with different
sizes
The particle size is an important tunable parameter in controlled
release system and definitely has a pronounced influence on the
release kinetics due to its different surface-area-to-volume ratios.
PME can easily adjust the size of the microcapsules in a large range
by simply changing the size of membrane pores, the viscosity of
the emulsion and the transmembrane pressure. Here, we adopted
three SPG membranes with pore sizes of 9.0, 7.2 and 1.0 ␮m to pro-
duce various LC-loaded microcapsules with sizes to be 4.6, 2.4 and
0.68 ␮m and LC contents to be 40.8%, 40.5% and 40.9%, designated
as MC1, MC2, and MC3 as indicated in Fig. 3. All the microcap-
sules had smooth surfaces and spherical shapes, and narrow size
distributions.
3.4. Controlled release of LC in vitro
In recent years, development of pesticide-release systems has
transited toward accurate and quantitative release from slow and
qualitative release. To achieve controllable and precise release, we
systematically investigated the release profiles of the LC-loaded
microcapsules with various particle sizes and pesticide contents.
Compared with LC technical and commercial microcapsule formu-
lation, all our microcapsules released LC in relatively slow speeds
and maintained its sustained release for longer periods (Fig. 4).
The LC technical was totally released after 18 h, and the cumula-
tive release of commercial microcapsule formulation reached 90%
44 B. Liu et al. / Colloids and Surfaces B: Biointerfaces 144 (2016) 38–45
in 12 h. Our microcapsules had an initial burst release in the first
25 h and then maintained a sustained and stable release. In the ini-
tial stage, the loaded pesticide at the surface of the microcapsules
would dissolve and leak into the surroundings quickly, resulting
in the burst release. It must note that the release profiles in vitro
obtained from an incubation medium of ethanol-water cannot pre-
cisely exhibit a practical release law in nature. Generally, when
the microcapsules are exposed to air or aqueous environments in
nature, the release rate of pesticides should turn slower, and initial
rapid release may be reduced. However, the various release profiles
of different drug delivery systems still can provide an insightful
understanding to evaluate their discrepancies in pesticide release,
especially for further release study in vivo. The effects of load-
ing content and the size of the capsules on the release of LC are
described in more detail below.
3.4.1. Effects of size on the LC release
We investigated the effects of size on the release profile of LC.
As shown in Fig. 4A, all the microcapsules had a slow release,
lasting for 250 h. As the size decreased from 4.6 nm to 0.68 ␮m,
the cumulative release increased from 67% to 82% after 48 h and
from 80% to 95% after 250 h. The results showed that the LC-
microcapsule delivery system with the smaller particle size had
the faster release of the active ingredient due to the higher surface
area being exposed to the surroundings, aiding the permeation and
effusion of the pesticide located in the shell of the microcapsule.
The results demonstrate change of particle sizes is an effectively
tunable way for the precise regulation of pesticide release.
3.4.2. Effects of the loading content on the LC release
In order to achieve a multi-regulated release system, the effects
of the LC loading content on the release behavior were further
investigated. The microcapsules with different LC contents were
obtained by changing the LC feeding content while maintaining the
same PLA concentration. As shown in Fig. 4B, the cumulative release
rates of LC increased from 78% to 95% after 250 h with increased
loading content from 22% to 45%. Higher LC contents in MC6 occupy
more spaces in the shells of the microcapsules. When the pesticide
molecules migrated and dissolved to the surroundings, more voids
in MC6 are formed, resulting in subsequent accelerated pesticide
release.
3.4.3. The release mechanism of the LC-loaded micocapsules
In order to investigate the release mechanism of the LC-loaded
microcapsules, the release data were analyzed by applying the
exponential relation proposed by Ritger and Peppas [46]:
M t /M 0 = kt n
where Mt /M0 is the percentage of active ingredient released at time
t, k is a constant, and n is the diffusion exponent. (The values of the
diffusion exponent n and T50 , the time taken for 50% of the active
ingredient to be released into water, were calculated and presented
in Table 2.) There was a good correlation between the release pro-
files of LC and the empirical equation with correlation coefficients
greater than 0.98. The lowest values of T50 for MC3 and MC6 indi-
cated the faster release of LC from suspensions of microcapsules
with smaller capsule sizes and higher loading contents.
The values of the diffusion exponent n were larger than 0.45,
indicating that the LC release from the microcapsules was con-
trolled by both a diffusion and matrix degradation. As depicted
in Fig. 4(C, D), the surface of the microcapsules after 250 h in the
medium became rough, and some pores appeared in the shells,
indicating the degradation of the microcapsules. The degradation
occurred because of the hydrolysis of the PLA on the surface.
3.5. Bioassay study of the LC-loaded microcapsules
To verify feasibility of the microcapsule suspension as a novel
pesticide formulation, bioactivity of LC in different microcapsules
against plutella xylostella was tested. Fig. 5 indicated the MC3
microcapsule (0.68 ␮m) had a lower LC50 value and exhibited obvi-
ously higher activity than the other two microcapsules, suggesting
that efficacy of pesticide gradually increased with the decrease of
the microcapsule size. The MC3 microcapsule had the similar effi-
cacy as a commercial formulation. The high efficacy should ascribe
to that the nano-sized microcapsules can enhance the adhesivity
and penetrability of pesticide on surface of crops, subsequently
reducing leaking loss of pesticide during spraying operation. How-
ever, compared with the commercial formulation, our LC loaded
microcapsules are more environmentally friendly because they
could be directly dispersed in water, avoiding the usage of toxic
organic solvents and a large number of pesticide adjuvants.
3.6. Stability of the LC-loaded microcapsules
3.6.1. UV-shielding properties of the microcapsules for LC
The photodegradation of pesticides after spraying seriously
destroys the activities of these pesticides and reduces their utiliza-
tion rates. This encapsulation technology is an important mean to
improve the optical stability of pesticides by incorporating pesti-
cides into capsule shells, preventing photodegradation and slowing
down the release rate of pesticide.
Fig. 6A showed that the decomposition rate of the encapsu-
lated LC was significantly reduced compared to that of LC technical.
The photolysis rate of LC technical was more than 11% after 12 h
of UV irradiation, while that of LC in the microcapsules was only
3%. Even after 72 h, only less than 18% of LC was degraded for
the microcapsules compared with up to 60% for the LC techni-
cal. The results clearly demonstrate that the microcapsules could
effectively reduce LC photolysis, indicating the microcapsule for-
mulation has the remarkable UV-shielding properties for LC.
3.6.2. Thermal-stability of the LC-loaded microcapsules
The storage stability is another key factor that affects the qual-
ity of a pesticide formulation. During storage, delamination, caking,
and degradation of the active pesticides may occur, reducing the
utilization of the pesticides and thus wasting resources. To deter-
mine LC thermal stability of the microcapsules, the samples were
stored at 4 ◦ C for 7 days and 54 ◦ C for 14 days. As shown in
Fig. 6(C–E), there were no obvious morphologic changes observed,
indicating that the microcapsules had good thermal stability. As
shown in Fig. 6B, the LC content only had a negligible loss after
storage at 4 ◦ C for 7 days. A small loss of LC was observed after
(1) 14 days at 54 ◦ C due to the degradation of LC at high temperatures.
The results showed that the solid microcapsules had good storage
stability.
4. Conclusion
In this work, we developed an environment-friendly controlled-
release delivery system for LC using biodegradable PLA as the
microcapsule shells. The LC-loaded microcapsule system had good
water dispersion and stability. Manipulating the process parame-
ters constructed various systems with LC contents as higher than
40% as well as tunable sizes from 0.68 to 4.6 ␮m. Compared with
both LC technical and commercial microcapsule formulations, the
LC-loaded microcapsule systems yielded LC sustained release for
a longer period. The LC release was controlled by LC diffusion and
matrix degradation. The 0.68 ␮m microcapsule system with good
UV and thermal stability had the similar efficacy as the commercial
formulation. We envision that such an environmentally friendly
 B. Liu et al. / Colloids and Surfaces B: Biointerfaces 144 (2016) 38–45
novel pesticide formulation should be greatly potential for wide
applications in the field of agriculture.
Acknowledgments
The authors thank the Major National Scientific Research
Program of China (Nos. 2014CB932200and 2014BAI11B04), the
National Young Thousand Talents Program and the National
Natural Science Foundation of China (NSFC, Nos. 21174147,
21474115and 51103165), and the Basic Scientific Research Fund
of the National Nonprofit Institutes for financial support.
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