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ABSTRACT: Emamectin benzoate is highly eective against insect pests and widely used in the world. However, its biological
activity is limited because of high resistance of target insects and rapid degradation speed in elds. Preparation and
physicochemical characterization of degradable microcapsules of emamectin benzoate were studied by modied solvent
evaporation/extraction method using polylactide (PLA) as wall material. The inuence of dierent compositions of the solvent in
internal organic phase and external aqueous phase on diameter, span, pesticide loading, and entrapment rate of the microspheres
was investigated. The results indicated that the process of solvent extraction and the formation of the microcapsules would be
accelerated by adding water-miscible organic solvents such as ethyl ether, acetone, ethyl acetate, or n-butanol into internal organic
phase and external aqueous phase. Accelerated formation of the microcapsules would result in entrapment rates of emamectin
benzoate increased to as high as 97%. In addition, by adding ethanol into the external aqueous phase, diameters would reduce to
6.28 m, whereas the loading eciency of emamectin benzoate did not increase. The PLA microspheres prepared under
optimum conditions were smoother and more spherical. The degradation rate in PLA microspheres of emamectin benzoate on
the 10th day was 4.29 0.74%, whereas the degradation rates of emamectin benzoate in methanol solution and solid technical
material were 46.3 2.11 and 22.7 1.51%, respectively. The PLA skeleton had combined with emamectin benzoate in an
amorphous or molecular state by using dierential scanning calorimetry (DSC) determination. The results indicated that PLA
microspheres of emamectin benzoate with high entrapment rate, loading eciency, and physicochemical characteristics could be
obtained by adding water-miscible organic solvents into the internal organic phase and external aqueous phase.
KEYWORDS: solvent evaporation/extraction method, emamectin benzoate, polylactide microspheres, physicochemical characteristics,
dierential scanning calorimetry
INTRODUCTION
Insect pests are the main limiting factor for vegetable
production in tropical Asia. Farmers often use synthetic
insecticides indiscriminately, and heavy insect resistance to
insecticides is very common in the elds. To control pests
eciently, some new pesticides, with novel modes of action,
have been developed recently. Among them, emamectin
benzoate is a novel insecticide deriving from the natural
product avermectin family with improved thermal stability,
greater water solubility, and a broader spectrum of insecticidal
activity than those of avermectin.1 Emamectin benzoate, with
novel modes of action similar to that of avermectin (GABAand glutamate-gated chloride channel agonist), high selectivity,
and greater safety to nontarget organisms, is highly eective
against lepidopteran pests and widely used in the world. It is
1880400-fold more potent against diamondback moth,
Plutella xylostella, cabbage looper, Trichoplusia ni (Hubner),
and beet armyworm, Spodoptera exigua (Hubner), than other
traditional insecticides, such as pronil, chlorfenapyr, and
tebufenozide.2 However, signicant resistance to emamectin
benzoate in target pests was found because of long-term heavy
usage of this insecticide.3 On the other hand, emamectin
benzoate was sensitive to light and had a rapid degradation rate
in natural conditions so that its biological activity was limited
2013 American Chemical Society
Received:
Revised:
Accepted:
Published:
12219
Article
Article
6000
7000
8000
9000
7000
7000
7000
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7000
7000
1.5%
1.5%
1.5%
1.5%
1.5%
1.5%
1.5%
1.5%
1.5%
1.5%
gelatin
gelatin
gelatin
gelatin
gelatin
gelatin
gelatin
gelatin
gelatin
gelatin
7000
1.5% gelatin
7000
7000
7000
7000
diameter (m)
DCM
DCM
DCM
DCM
DCM/ether (1:1)
DCM/n-butanol (1:1)
DCM/acetic ether(1:1)
DCM/acetone(1:1)
DCM/acetone (1.5:0.5)
DCM/acetone
(1.25:0.75)
DCM/acetone
(0.75:1.25)
DCM
DCM
DCM
DCM
14.4
11.6
9.02
6.02
17.2
9.77
13.4
16.5
12.2
14.8
1.09
0.51
0.59
0.57
0.64
0.98
1.10
1.41
0.74
0.44
fg
e
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a
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d
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hi
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fg
15.4 0.47 gh
10.2
9.04
7.99
6.82
0.38
0.36
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d
cd
bc
ab
pesticide loading
(%)
span
1.50
1.45
1.44
1.40
1.47
1.55
1.53
1.49
1.24
1.44
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0.03
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bcd
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26.0
24.4
22.4
19.7
24.2
23.5
28.0
31.8
25.9
29.4
0.34
0.78
0.58
0.36
0.82
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0.73
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def
d
b
a
entrapment rate
(%)
79.2
74.3
68.2
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73.8
71.6
85.3
97.0
79.0
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1.02
2.37
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84.3 2.22 hi
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52.1
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1.34
1.34
3.82
2.12
def
d
b
a
DCM, dichloromethane; EtOH, ethanol. Dierent lower case letters indicate signicant dierence (P < 0.05).
Figure 2. Eects of internal organic phase on the quality of microcapsules. In control, dichloromethane alone was used. In other treatments, the
dichloromethane was mixed with ether, n-butynol, ethyl acetate, or acetone at the ratio of 1:1, respectively.
methanol/acetonitrile/triethylamine solution (0.1%, 50:40:10), with a
ow rate of 1.0 mL/min. The detection wavelength was 245 nm, and
the injection amount was 20 L. From the concentration, the
entrapment rate and content of emamectin benzoate in the prepared
microspheres were calculated.
Mean Diameter and Span. Amounts of 0.25 g of microspheres and
0.2 g of sodium phosphate were dissolved with 30 mL of distilled
water under ultrasonic condition. The mean diameter was measured
using an LS-POP(7) laser particle size analyzer (OMEC Instrument
Co., Ltd., Zhuhai, China) after high-speed sample introduction. Span =
(D90 D10)/D50. D50 represents the mean diameter of microspheres,
D90 represents the diameter of 90% of the microspheres, and D10
represents the diameter of 10% of the microspheres.
Dierential Scanning Calorimetry (DSC). Ten milligrams of four
samples, including emamectin benzoate, PLA microspheres of
emamectin benzoate, a physical mixture of emamectin benzoate and
blank PLA microspheres, and blank PLA microspheres without
emamectin benzoate, was used for analysis by DSC. Thermal analyses
RESULTS
Eects of Emulsifying Speed on Microsphere Characteristics. The mean diameter and the entrapment rate
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Article
Figure 3. Scanning electron micrographs of microspheres prepared using dierent internal organic phases: (A, C) internal organic phase was the
mixture of dichloromethane with acetone (1:1); (B, D) internal organic phase was dichloromethane alone.
Figure 4. Eects of acetone content in internal organic phase on the quality of microcapsules.
Article
Figure 5. Eects of ethyl alcohol content in external aqueous phase on the quality of microcapsules.
Figure 6. DSC spectrogram diagram (left) and thermal weight loss (right): a, emamectin benzoate; b, PLA microspheres of emamectin benzoate; c,
physical mixture of emamectin benzoate and blank PLA microspheres; d, blank PLA microspheres without emamectin benzoate.
Article
DISCUSSION
It was known that emulsifying speed was the main parameter
for controlling the dispersions droplet size and directly aected
the diameter of microspheres under the existence of sucient
emulsier condition.16 Our results indicated dierent diameters
of microspheres could be obtained by changing the emulsifying
speed according to actual needs in experiments. With the
increase of emulsifying speed, the dispersion and the specic
surface area of organic emulsion droplets in continuous phase
increased, but the organic emulsion droplet sizes decreased.
The facts resulted in the diusion probability of the emamectin
benzoate to external aqueous phase increased and the
entrapment rate decreased. Generally speaking, higher
emulsifying speed and stronger shear force would result in
better dispersion eect of dispersed organic phase in external
aqueous phase and smaller particle size of microspheres.30
Therefore, in the preparation process of emamectin benzoate
microspheres by using the emulsion solvent evaporation/
extraction method, the emulsifying speed was the critical factor
that aected the emulsion droplet size and nally inuenced the
size and entrapment rate of microspheres.
Bodmeier et al. reported that the entrapment rate of drug
increased when dichloromethane was mixed with organic
solvents such as acetone, methanol, ethanol, dimethyl sulfoxide,
and ethyl acetate in the preparation of poly(D,L-lactide)
microspheres containing quinidine or quinidine sulfate
prepared by the solvent evaporation method.26 Niwa et al.
reported that nanospheres with higher entrapment rate and
lower size could be obtained when a mixture of acetone and
dichloromethane (or acetone and chloroform) was used as
organic solvent in the preparation of PLGA nanospheres
containing indomethacin and 5-uorouracil, and the diameter
of nanospheres decreased when the amount of acetone in
internal organic phase increased.25 In this paper, the entrapment rate of emamectin benzoate increased when a suitable
amount of acetone or ethyl acetate was added into the internal
organic phase. However, dierent from the results above,25,26
the diameters of microspheres increased when the amount of
acetone in internal organic phase increased. The increase of
microphere diameter might be related to the fact that the
formation speed of microspheres was so fast that microspheres
gathered together easily. On the other hand, microspheres with
high entrapment rate and suitable diameter could be also
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Article
ASSOCIATED CONTENT
S Supporting Information
*
AUTHOR INFORMATION
Funding
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