Colloid and Polymer Science

https://doi.org/10.1007/s00396-020-04664-x

ORIGINAL CONTRIBUTION

Preparation and application performance of lignin-polyurea

composite microcapsule with controlled release of avermectin
Yuxia Pang 1 & Zhicai Qin 1 & Shengwen Wang 2 & Conghua Yi 1 & Mingsong Zhou 1 & Hongming Lou 1,3 & Xueqing Qiu 1,4

Received: 4 November 2019 / Revised: 15 February 2020 / Accepted: 25 March 2020

# Springer-Verlag GmbH Germany, part of Springer Nature 2020

Abstract
Pickering emulsion stabilized by lignin/sodium dodecyl sulfate composite nanoparticles (LSNP) was used as template to prepare
the avermectin @ lignin/polyurea composite microcapsules (AVM@LPMC) through ion cross-linking and interfacial polymer-
ization. The inner wall of the microcapsules is a firm and compact polyurea layer, and the outer wall is a loose lignin layer. The
effects of stirring speed, dosage of sodium dodecyl sulfate (SDS), and pH value in aqueous phase on the formation of micro-
capsules were systematically studied. The results showed that the optimal stirring speed was 200 rpm, and the optimal dosage
ratio in water phase was HCl (mmol):SDS (mmol):lignin AL (g) = 0.375:1.25:1 in fixed oil-water ratio (1:9) and oil phase
composition. In this way, the encapsulation efficiency of microcapsules could reach up to 85.4%, while it would slightly decrease
with the increase of lignin content in the wall materials. The polyurea layer played a key role in supporting the spherical structure
of the capsule wall and delaying the release of avermectin, while the loose lignin layer contributed less to the slow release
performance of microcapsule. By changing the amount of lignin, the polyurea-layer thickness could be regulated to adjust the
release rate of microcapsule. Remarkably, a small amount of lignin introduced in the wall material could significantly improve the
anti-photolysis performance of avermectin in microcapsules.

Keywords Lignin . Polyurea . Microcapsules . Pickering emulsion . Avermectin

Introduction play a variety of roles, such as changing the state or surface

Microcapsule, a kind of micro-container with cored-shell
structure whose size ranges from a few microns to several
hundred microns, could be used to cover various materials
including solid, liquid, and gas. This core-shell structure could
* Hongming Lou
cehmlou@scut.edu.cn
* Xueqing Qiu
xueqingqiu66@163.com
1
School of Chemistry and Chemical Engineering, Guangdong
Engineering Research Center for Green Fine Chemicals, South China
University of Technology, Wu Shan Road, Guangzhou 510640,
China
2
Guangzhou Liby Enterprise Group Limited Company,
Guangzhou 510370, China
3
State Key Laboratory of Pulp and Paper Engineering, South China
University of Technology, Guangzhou, China
4
School of Chemical Engineering and Light Industry, Guangdong
University of Technology, Guangzhou, China
properties of substances, protecting materials from destruc-
tion, controlling the release of substances and isolating com-
ponents [1]. Therefore, it has been widely applied in various
fields including medicine, cosmetics, food, pesticides, dyes,
and so on [2, 3].
At present, the walls of microcapsules are usually made of
synthetic polymers (such as polyurea [4], polyurethane [5],
polyamide [6], urea-formaldehyde resin [7], melamine form-
aldehyde resin [8]) by interfacial or in-situ polymerization and
natural polymers (mainly including proteins like gelatin [9],
carbohydrates like arabic gum [10], cellulose [11], chitosan
[12], sodium alginate [13] and so on) through physicochemi-
cal method. However, most of the wall materials mentioned
above are not effective enough to protect sensitive core mate-
rials that are easily degraded by light or oxidation.
Lignin, the second largest renewable resource after cellu-
lose in plants [14], is an amorphous polymer containing three
phenylpropanoid monomers (monolignols) linked by carbon–
carbon and ether bonds [15].Based on the large number of
benzene ring, abundant conjugated structure and phenolic hy-
droxyl which are effective to scavenge free radicals, lignin has

excellent thermal stability [16], ultraviolet absorption [17],
and antioxidant activity [18]. Therefore, compared with other
natural polymer, lignin exhibited unique advantage as micro-
capsule wall materials since it could significantly enhance the
light stability [19] of photosensitive or oxygen sensitive core
materials like pesticide avermectin (AVM) or curcumin [20,
21], which is extremely easy to decompose under UV irradi-
ation. Here, AVM is a kind of compound containing a
macrolides and conjugated double bonds that has strong par-
asite killing activity and usually used as a kind of low toxicity,
broad spectrum, and high efficiency biological insecticide.
Lignin microcapsules were prepared followed by a high-
intensity, ultrasound-assisted cross-linking of lignin at the
water/oil interface by Claudia Crestini [22]. However, the
cross-linking effect by ultrasound is not strong enough, and
the microcapsule shell is thin and easy to break. Deng [23] and
Li [24] respectively used different kind of lignin to prepare
colloidal spheres encapsulating avermectin through self-
assembly method and found that lignin could effectively
weaken the degradation effect of ultraviolet light on
avermectin and significantly prolong the existence time of
avermectin under light conditions. Besides, Liu [25] also pre-
pared lignin microcapsules with good slow-release properties
and UV-shielding properties through this way. However, the
method of self-assembly is complex and the operating con-
centration as well as the yield is low. Specially, microcapsules
prepared in this way exhibited poor mechanical performance
due to the relative thin capsule wall. All of these limited its
application in industry. Remarkably, traditional synthetic
polymer microcapsules like polyurea microcapsules have ad-
vantages of good film-forming and high chemical stability
[26]. Therefore, exploring a facile, low-cost and environmen-
tally friendly strategy to prepare composite-wall material mi-
crocapsules consisting of lignin and traditional synthetic poly-
mer and having both satisfactory shell strength, slow release
and anti-photolysis properties, has become a research hotspot.
We have reported a method of preparing novel lignin/SDS
composite nanoparticles (LSNPs) and its preliminary applica-
tion in making lignin/polyurea composite microcapsules
(LPMCs) previously [27]. In this method, novel lignin/SDS
composite nanoparticles (LSNPs) were facilely prepared by
acidifying the mixed solution of alkaline lignin and SDS.
The oil-in-water Pickering emulsion was further prepared by
using the LSNP aqueous dispersion as the water phase and
cyclohexane solution containing IPDI as the oil phase. Using
the emulsion as the template, LPMCs were prepared by
adding ethylenediamine to conduct ion-cross-linking reaction
and interfacial polymerization. As microcapsules for encapsu-
lating pesticide, LPMCs could endow pesticide with good
sustained release performance, long duration of efficiency as
well as improved anti-photolysis property. As mentioned
above, AVM is an extensively used biological insecticide with
low toxicity, broad spectrum and high efficiency. However,
Colloid Polym Sci
AVM is of great photosensitive and oxygen sensitive, leading
to its easy degradation under UV irradiation and application
limit. Therefore, it would be significant to encapsulate AVM
with suitable microcapsules to improve its anti-photolysis per-
formance and achieve sustained release.
On this basis, for further studying the preparation technol-
ogy and performance of LPMCs, revealing the formation
mechanism of LPMCs as well as exploring the application
of LPMCs in encapsulating pesticide AVM to achieve
sustained release and anti-photolysis properties, we prepared
the microcapsules AVM@LPMC by dissolving IPDI and pes-
ticide AVM in toluene as oil phase. Through monitoring mor-
phology and particle size, the preparation process of lignin/
polyurea composite microcapsule was specifically studied and
an appropriate proportion of acid, SDS, and lignin in the re-
action system was found. By studying the release behavior of
AVM@LPMC with different lignin dosage, the contributions
of polyurea and lignin to the sustained release performance of
microcapsule were revealed. Besides, the effect of the amount
of lignin on the photolysis resistance of the encapsulated sub-
stance was also studied. The results could provide more de-
tailed theoretical datas for the practical application of the com-
posite microcapsules.
Experiment section
Materials
Alkali lignin (AL), from Hunan Xiangjiang Paper Industry
Co., Ltd., was obtained by acidification of the pine alkali
pulp-making black liquor and filtration with ultrapure water
and used without other modifications. The purity was about
90%, with the impurities mostly being organic acids, sugar,
low molecular and carbohydrates. The AVM powder (OP-
AVM, 92.2% w/w) was provided by Noposion Co., Ltd.
(Shenzhen, China). Hydrochloric acid, sodium hydroxide,
polyvinyl alcohol (PVA), and toluene were all purchased from
Guangzhou Chemical Factory (China). Isophorone
diisocyanate (IPDI), ethylenediamine (EDA), and sodium do-
decyl sulfate (SDS) were purchased from Shanghai Macklin
Biochemical Technology Co., Ltd. (China). All of the reagents
were analytical-grade and used without further purification.
HPLC grade methanol was purchased from Honeywell
International Co., Ltd. (Tianjin, China).
Experimental procedures
Preparation of LSNPs
In our previous study [27], lignin/SDS composite nanoparti-
cles (LSNPs) with size ranging from 70 to 200 nm were suc-
cessfully prepared via acidification of the mixed micelles,
Colloid Polym Sci
which were formed by the self-assembly of lignin and SDS.
Since the preparation and characterization of LSNPs have
been fully studied previously, in this work, LSNPs were pre-
pared according to our existing method as follows: the lignin
particles are first dissolved with SDS in an alkaline aqueous
solution to make them self-assemble, and then the key step is
to adjust the pH by adding HCl to precipitate the lignin com-
posite nanoparticles. The specific operation is as follows: first,
0.8, 1.0, 1.3, 1.5 g alkali lignin were respectively added into
90 mL ultrapure water, after which 4 mol/L NaOH was added
under continuous agitation until the pH of the lignin solutions
reached 11, and the impurities were removed by filtration.
Later, a certain amount of SDS was added and fully stirred
to obtain the mixed solution of lignin/SDS. Finally, LSNP
dispersions with different lignin dosages were prepared by
adjusting the pH of the mixed solution to 2–3 with 1 mol/L
HCl under the agitation of 400 rpm.
Preparation of AVM@LPMC and AVM@PMC
Firstly, 0.6 g AVM and 3.6 g IPDI were dissolved in 10 mL
toluene to form the mixed oil phase. After mixing the LSNP
dispersion with the oil phase, the system was immediately
emulsified at 11,000 rpm for 3 min with a homogenizer
(IKA Ultra Turrax T25 basic instrument) to obtain Pickering
emulsion. Then, the emulsion was transferred to a water bath
at 35 °C. With a stirring speed of 150~300 rpm, 10 mL
1.5 mol/L EDA solution was added dropwise for 1 h, after
which the reaction was continued for 5 h under stirring to get
lignin/polyurea composite microcapsules (AVM@LPMC).
Figure 1 illustrates the formation of AVM@LPMC. Part of
LSNPs in the aqueous phase was adsorbed on the emulsion
drops interface and formed the oil-in-water Pickering
Fig. 1 Illustration of the
preparation of LPMC
emulsion stably. The HCl in the aqueous phase reacted with
the added ethylenediamine to form ethylenediamine hydro-
chloride. The ionized ethylenediamine hydrochloride divalent
cations (EH ion) could undergo cross-linking reaction with the
negatively charged LSNPs on the surface of the emulsion
droplets to form the lignin microcapsules. Meanwhile, the
decrease in the repulsion forces permitted LSNPs, which were
cross-linked in aqueous phase to deposit onto the surface of
the microcapsules. After the HCl in the aqueous phase was
exhausted, the continually added EDA would react with IPDI
in the oil-water interface to form a polyurea layer. Eventually,
a composite microcapsule whose capsule wall was made of
smooth polyurea layer. With the ion-cross-linked lignin gran-
ules attached outside was formed.
The avermectin@polyurea microcapsules (AVM@PMC)
were prepared similarly to the preparation process of
AVM@LPMC except that LSNP dispersion was replaced by
90 mL of deionized water dissolved with 0.45 g PVA.
Characterization
Structural analysis of alkali lignin
The functional group of alkali lignin was determined by non-
aqueous conductometric titration using the automatic
porentiometric titrator (809 Titrando, Metrohm Corp.,
Switzerland). The molecular weight dates of alkaline lignin
were measured by gel permeation chromatography (Agilent
1100 series, Agilent, USA) with PLgel 5 μm 1000 Å and
PLgel 5 μm 500 Å columns. The tetrahydrofuran was set as
mobile phase, and the flow rate was set at 1 mL min−1. The
effluent was monitored using a G1362 Å RID detector. Before

test, the instrument was calibrated by using polystyrene (PS)
as the standard substance [28].
The results showed that the phenolic OH and carboxylic of
alkali lignin were 2.18 mmol/g and 2.00 mmol/g, respectively,
and the Mw as well as Mn was 4500 Da and 2470 Da,
respectively.
Determination of lignin loading in the wall of microcapsule
After keeping the reaction mixture standing for a while, the
prepared microcapsules would float to the surface and the
unbound lignin would sink. The microcapsules were poured
out and the remaining lignin was filtered, dried, and weighted.
And the mass of lignin combined in the microcapsule was
calculated as the difference between the initial added mass
and the remaining one. The capsule-formation ratio of lignin
on capsule wall was defined as the ratio of the mass of com-
bined lignin to the initial added mass.
Size analysis
The size distributions of emulsion and microcapsule were
studied using a particle size analyzer (Mastersizer 2000,
Malvern Instruments, UK) equipped with a Hydro EV wet
dispersion unit. Before the test, the sample was placed in the
sample pool and diluted with ultra-pure water until the shad-
ing rate reached 15%, and the mixing speed of the sampler
was set at 2000 rpm.
FTIR spectra
Fourier infrared spectrometer (Thermo Nico Let 380, Thermo,
detection wavelength 400–4000 cm−1) was used to analyze
the composition of the lignin, IPDI, capsule walls of micro-
capsules. Before the test, all microcapsule samples were
crashed by ultrasonic first, then the resultant slurry was rinsed
with ethanol and distilled water three times to remove the
AVM, toluene and unreacted EDA, and finally, the treated
samples were vacuum-dried at 50 °C for 24 h. All samples
were mixed with potassium bromide (1/99, w/w) and pressed
under the pressure of 10 MPa for 5 min before FTIR testing.
Microscopic observation
Emulsion or microcapsule samples were observed and
photographed with a polarizing microscope (BM-5000P,
Shanghai bimu). Before observation, the emulsion or microcap-
sule dispersion solution should be diluted. The morphological
characteristics of microcapsules were observed by CLSM
(Leica TCS-SP5, Leica, Germany, excitation light source
wavelength 405 nm) and field mission SEM (LEO1530VP).
Colloid Polym Sci
Encapsulation ratio and drug loading
The mass ratio between the pesticide encapsulated in micro-
capsules and the initial added one was defined as the encap-
sulation ratio. Drug loading refers to the mass percentage of
pesticide in the dry microcapsule.
One milliliter of microcapsule suspension which was fully
shaken was added into 50 mL of anhydrous ethanol, and two
parallel samples were prepared as follows. One sample was
ruptured by ultrasound for 1 min to ensure the AVM fully
dissolved in ethanol. Then, the relative concentration C0
(mg/L) of AVM was determined by HPLC after filtration.
The other sample was shaken and filtered to obtain microcap-
sule solid and filtrate. The microcapsules were dried at room
temperature and weighed as M0 (mg), and the relative concen-
tration C1 (mg/L) of AVM in the filtrate was detected. The
encapsulation ratio, θ (%), could be calculated according to
the Eq. (1), and the drug loading of φ (%) could be calculated
according to Eq. (2):
 
C1
θð%Þ ¼ 1−  100% ð1Þ
C0
50*ðC 0 −C 1 Þ
φð% Þ ¼  100% ð2Þ
M0
Determination of AVM concentration: using high perfor-
mance liquid chromatography (HPLC, Agilent ZORBAX,
Agilent, USA) with isocratic elution of methanol-water (88/
12, v/v) as the mobile phase, the injection volume was 20 μL
and separated at 40 °C, using a flow rate of 1 mL/min under
the detection wavelength of 245 nm, which is the maximum
absorption peak of AVM in the UV spectra [23]. Each sample
was tested 3 times and averaged.
Sustained release of microcapsules
AVM release from the microcapsules was evaluated through a
dynamic dialysis method. One milliliter of microcapsule sus-
pension was accurately measured, diluted in 20 mL ethanol/
water (80/20, v/v) mixed solution, and then transferred into a
dialysis bag with interception molecular weight of 10,000 D.
The dialysis bag was immersed in 100 mL ethanol/water (80/
20, v/v) mixed solution and continuously shaken in a shaking
bed at 25 °C and 200 rpm. At certain intervals, 1 mL solution
outside the dialysis bag was taken out to test the AVM con-
centration, and meanwhile, the same volume of ethanol/water
(80/20, v/v) mixed solution was added to maintain a constant
volume.
In order to evaluate the sustained release of AVM-loaded
microcapsules, the cumulative release rate of AVM-loaded
microcapsules was calculated by measuring the concentration
of AVM dissolved in the mixture solution. Time was taken as
the horizontal coordinate and the cumulative release
Colloid Polym Sci

percentage of AVM as the vertical coordinate to draw the Table 1 The influencing factors of microcapsule formation
release curve of AVM. Meanwhile, AVM toluene solution, Reactions SDS (g) pH Speed (rpm)
whose concentration and volume were the same as AVM in
microcapsule oil phase, was set as the control group, and the 1 0.36 2.50 150
sustained release experiment was conducted under the same 2 0.36 2.50 200
operation condition. 3 0.36 2.50 250
4 0.36 2.50 300
Photodegradation of AVM-loaded microcapsules 5 0.20 2.50 m
6 0.22 2.50 m
One milliliter of microcapsule suspension was diluted in 5 mL 7 0.28 2.50 m
deionized water, and 6 samples were prepared in parallel. 8 0.36 2.50 m
They were placed under ultraviolet lamp (16 W, 310 nm) 9 n 2.00 m
and irradiated for 6 h, 12 h, 24 h, 36 h, 60 h, and 96 h, respec- 10 n 2.25 m
tively. After the irradiation, the corresponding samples were 11 n 2.50 m
taken out and treated with ultrasound to ensure the complete 12 n 3.00 m
release of AVM. And the concentration of AVM was deter-
mined by HPLC after adding anhydrous ethanol to ensure the m, n: the optimal values of agitation speed and SDS dosage, respectively
constant volume of 50 mL. According to the detection results
of 6 samples, the light degradation curve was plotted with the
irradiation time as the horizontal coordinate and the retention For reactions of 1~4 in Table 1, the effects of different
percentage of AVM as the vertical coordinate. Meanwhile, the stirring speeds were investigated under the condition of fixed
photodegradation curve of free AVM was plotted with the SDS dosage of 0.36 g and pH of 2.5, and the microscope
control group of AVM ethanol solution, whose concentration images of LPMC are shown in Fig. 2. Most microcapsules
and volume were the same as AVM in microcapsule oil phase. prepared at 150 rpm and 200 rpm were regular spheres, espe-
cially those prepared at 200 rpm exhibited more regular mor-
phology and smaller particle size. However, at the speed of
Results and discussion 250 rpm and 300 rpm, the emulsion droplets or microcapsules
were subjected to violent collisions, causing rupture or coa-
Preparation conditions of microcapsules LPMC lescence and irregular shapes with larger particles. Therefore,
200 rpm is the optimal stirring speed for the preparation of
Single-factor method was adopted to study the preparation microcapsules.
process of lignin/polyurea composite microcapsules. That is
the remaining conditions were controlled constant (90 mL
water, 10 mL toluene, 1 g lignin, 2.4 g IPDI, 0.6 g AVM, SDS dosage
10 mL 1.5 mol/L EDA, and the emulsification speed was
11,000 rpm), and the stirring speed, SDS dosage, and pH Pickering emulsion was prepared by SDS-doped lignin nano-
value of acidifying lignin particle were respectively changed particles. As the amount of SDS increased, the particle size of
to prepare microcapsules. The optimum value of every factor nanoparticles decreased significantly, which would have an
was obtained by comparing the microcapsule morphology, impact on the particle size of emulsion droplets formed by
and the specific operating conditions are shown in Table 1. emulsification, and thus affected the size of microcapsules.
When the pH value was set at 2.5 and the stirring speed was
Stirring speed 200 rpm, the reactions of 5~8 was carried out according to
Table 1, and the influence of SDS dosage on the formation of
On the one hand, stirring could fully disperse the emulsion emulsion and microcapsule was investigated. Figures 3 and 4
system, prevent the agglomeration of emulsion droplets in the illustrate the microscope images and particle size distribution
polymerization process, and facilitate the mass transfer of the curves of Pickering emulsion and microcapsules prepared un-
reaction system that improved the utilization efficiency of der different SDS dosage, respectively. It could be observed
monomer and accelerated the formation of capsule wall. On that the emulsion droplets were all in regular spherical shape,
the other hand, excessive agitation would increase the energy and the particle size gradually decreased with the increase of
of the system and thus increase the probability of collision SDS dosage. This phenomenon indicated that the LSNPs with
between emulsion droplets, which would aggravate the coa- smaller particle size formed under larger SDS amount [27],
lescence between droplets, thus leading to demulsification and and abundant free SDS in the aqueous phase play a synergistic
hindering the formation of microcapsules. role in emulsification.

Fig. 2 Optical microscopy
images of microcapsules prepared
at different stirring speed
The particle size of emulsion template largely determined
the size of microcapsules, while the particle size of microcap-
sule was significantly larger than that of emulsion template,
and the distribution was bimodal, as shown in Fig. 4. This was
because ethylenediamine hydrochloride ions would be gener-
ated in the process of forming microcapsules, and this divalent
cation caused serious damage on the stability of Pickering
emulsion stabilized by negative charged SDS and LSNP [29,
30]. Under the mechanical agitation condition, emulsion drop-
lets would aggregate to form droplets with larger particle size,
which led to the formation of larger microcapsules. At the
same time, the free LSNP would also be cross-linked by
ethylenediamine hydrochloride ions to form aggregates
scattered in the water, resulting in the part of small particle
in bimodal distribution.
Volume (%)
Fig. 3 Optical microscopy images (left) and size distribution (right) of Pickering emulsion prepared with different amount of SDS
Colloid Polym Sci
As the optical microscopy images of microcapsules shown
in Fig. 4 (left), the particle size of microcapsules first de-
creased and then increased with the increase of SDS dosage.
This could be attributed to two aspects. On the one hand, the
emulsification of SDS reduced the particle size of the emul-
sion and the corresponding microcapsules. On the other hand,
the excessive negative charge of SDS made the emulsion
droplets easily influenced by the divalent cation of
ethylenediamine hydrochloride, leading to demulsification
and droplet enlargement. Under the SDS dosage of 0.36, the
demulsification and increase of particle size was more obvi-
ous. Therefore, the microcapsules reached the minimum size
when the SDS dosage was 0.28 g.
However, according to the data measured from a particle
size analyzer, the maximum particle size was found when the
14
0.20 g SDS
0.22 g SDS
12 0.28 g SDS
0.36 g SDS
10
0
0.1 1 10 100
Particle Size (µm)
Colloid Polym Sci
Volume (%)
Fig. 4 Optical microscopy images (left) and size distribution (right) of microcapsules prepared with different amount of SDS
SDS dosage was 0.28 g, shown in Fig. 4 (right), which was
totally different from the results of the optical microscopy
images. The actual situation was that when the dosage of
SDS reached 0.28 g, the size of microcapsules was so small
that led to a large concentration of microcapsules. In a rela-
tively high concentration, microcapsules were easy to form
large microcapsule cluster through cross-linking by
ethylenediamine hydrochloride ions, which has been proved
and circled in red in Fig. 4 (left), and thus showed the maxi-
mum size in a particle size analyzer. Moreover, when the
dosage of SDS reached 0.36 g, the concentration of microcap-
sules was lower compared with the situation where the dosage
of SDS was 0.28 g. Meanwhile, the negative charges of
LSNPs which were derived from SDS led to the electrostatic
repulsion between microcapsules. Therefore, in the case when
the SDS dosage was 0.36 g, it was relatively difficult for
microcapsules to form clusters as the SDS dosage was
0.28 g. That was why the SDS dosage of 0.36 g did not induce
larger particle size in comparison with that of 0.28 g from the
results of particle size analyzer.
In terms of morphology, microcapsules appeared as regular
spheres with a rougher surface than that of emulsion droplets,
and the larger amount of SDS was used, the darker the micro-
capsules were, indicating that increasing the SDS dosage is
beneficial to the effective utilization of lignin on the wall of
microcapsules. Therefore, in order to fully load lignin into
microcapsules, the mass ratio of SDS to lignin was determined
to be 0.36:1, that is, SDS (mmol):AL (g) = 1.25:1.
The pH of the water phase
Under the condition of fixed SDS dosage of 0.36 g and stirring
speed of 200 rpm, microcapsules were prepared at different
pH values shown as the reactions of 9–12 in Table 1 and the
microscope images are described in Fig. 5. As is shown, the
pH value of aqueous phase significantly influenced the
14
0.20 g SDS
0.22 g SDS
12
0.28 g SDS
0.36 g SDS
10
8
0
0.1 1 10 100
Particle Size (µm)
particle size and morphology of microcapsules. The pH value
of the aqueous phase reflected the concentration of HCl in the
aqueous phase. According to the formation mechanism of the
microcapsules, the lower the pH value of the aqueous phase,
the higher the HCl concentration in the aqueous phase, and
thus the more ethylenediamine hydrochloride ions would be
generated after the addition of EDA. On the one hand,
ethylenediamine hydrochloride ions caused the interfacial
LSNPs of the emulsion droplets to be ionized cross-linked to
form the capsule wall. On the other hand, excessive
ethylenediamine hydrochlorides led to the coalescence be-
tween emulsions and excessive cross-linking of LSNP,
resulting in the increasing particle-size of prepared microcap-
sules and severe agglomeration between microcapsules.
Therefore, irregular aggregations were formed of microcap-
sules at pH = 2.0 or 2.25, and microcapsules prepared at pH
values of 2.5, 2.75, and 3.0 have regular morphology, and with
the increase of pH, the particle size of microcapsules becomes
smaller and the surface becomes smoother.
In the preparation process of microcapsules, the higher the
concentration of SDS, the larger amount of SDS was bound in
LSNP, and the more HCl was required to provide more
ethylenediamine hydrochloride ions to cross-link LSNP. As
shown in Fig. 5, the appropriate pH of aqueous phase was
2.5, and it could be calculated and detected that the optimal
HCl dosage was 0.375 mmol. Therefore, considering the op-
timal dosage ratio of lignin and SDS mentioned above, the
optimal ratio of HCl:SDS:lignin in 90 mL aqueous phase was
HCl (mmol):SDS (mmol):AL (g) = 0.375:1.25:1 when the lig-
nin dosage was set at 1 g.
Lignin dosage
A series of microcapsules were prepared with different
lignin content (0.8, 1.0, 1.3, and 1.5 g, respectively) in
the water phase, which oil phases were composed of

Fig. 5 Optical microscopy images of microcapsules prepared with different aqueous pH
0.6 g AVM and 2.4 g IPDI dissolved in 10 mL toluene
and the oil-water ratios were fixed at 1:9, and the amount
of SDS and HCl was set as the above optimal feeding
proportion. The microscope images of microcapsules
were shown in Fig. 6. It could be seen that the particle
size of microcapsules gradually increased with the in-
crease of lignin dosage, and the amount of lignin bound
on the surface of microcapsules also increased, which
was reflected on the darker color and rougher surface.
Figure 7 shows the mass of combined lignin in mi-
crocapsules and the corresponding capsule-formation ra-
tio at different lignin dosage. As the lignin dosage in-
creased, the amount of lignin in the capsule wall
Fig. 6 Optical microscopy
images of microcapsules with
different weight of lignin
Colloid Polym Sci
increased from 0.61 g to 0.93 g, but the capsule-
formation ratio decreased from 84.7 to 68.8%, indicat-
ing that it was more difficult for lignin to continue
combining onto the capsule wall. Besides the cross-
linking effect of ethylenediamine hydrochloride ions,
the reaction between IPDI in oil phase and hydroxyl
groups in lignin also contributed to the cross-linking
of lignin, resulting in firmer and tighter lignin-polyurea
layer of microcapsules. When a certain amount of lignin
was combined on the surface of emulsion droplets, it is
difficult for the excess lignin to contact with the IPDI
in the oil phase and to firmly attach on the outer layer
of capsules.
Colloid Polym Sci

Fig. 7 Amount and capsule- 1.1 100

formation ratio of lignin on cap-
1.0 90
sule wall as a function of the

Percentage of lignin on capsule wall (%)

Amount of lignin on capsule wall (g)
amount of added lignin 0.9 80
0.8
70
0.7
60
0.6
50
0.5
0.93 40
0.4 0.84
0.73
30
0.3 0.61
0.2 20

0.1 10

0.0 0
0.8 1.0 1.3 1.5
Amount of added lignin (g)

Morphology and composition characterization
of microcapsules
LPMC prepared with 1 g lignin content was observed with a
scanning electron microscope and a laser confocal microscope,
as shown in Fig. 8. In order to determine the stable existence of
lignin in the microcapsule wall, the LPMC was soaked in
0.5 mol/L NaOH solution and shaken for 2 h before observation
to remove the lignin on the surface of microcapsules.
The SEM images (Fig. 8a) showed that the morphology of
LPMC was regular. Outside of the smooth and dense shell
wall, there were large number of granular substances attached.
The smooth layer was polyurea capsule wall and the granules
which embedded independently on the surface of the smooth
capsule wall were ion-cross-linked LSNPs. From Fig. 8b, the
cross-sectional SEM image of LPMC, it can be found that the
smooth capsule wall was about 90 nm. The thin capsule wall
made it difficult for the microcapsule without core material to
bear the lignin granules on the outer surface, so the capsule
wall easily deformed.
Because of the large amount of conjugate structural units in
lignin, which could emit blue-green fluorescence under the
excitation of ultraviolet light [31, 32], the distribution of lignin
in LPMC could be observed by laser confocal microscope. As
shown in Fig. 8c, LPMC has a very strong fluorescence profile
on the surface after alkali treatment, confirming that large
number of lignin was cross-linked onto the microcapsule wall
due to the reaction between lignin and IPDI.

Fig. 8 Scanning electron

microscope (a, b) and laser
confocal microscope (c) images
of LPMC

Fig. 9 FT-IR spectra of AL, IPDI, capsule wall of PMC and LPMC
Lignin, IPDI, capsule walls of AVM@LPMC and
AVM@PMC were characterized by infrared spectroscopy
and the results were shown in Fig. 9. In PMC capsule wall,
the absorption peak at 3350 cm−1 was attributed to the
stretching vibration of N-H in amide group, and the peak at
2950 cm−1 was ascribed to stretching vibration of C-H of
methyl and methylene groups in IPDI. Moreover, the asym-
metric stretching vibration absorption peak of N=C=O in IPDI
at 2250 cm−1 was very weak, indicating the successful reac-
tion between IPDI and ethylenediamine to generate polyurea.
Compared with the PMC capsule wall, strong absorption
peaks also appeared at 3350 cm−1 and 2950 cm−1 in the infra-
red spectrum of LPMC capsule wall, confirming the existence
of polyurea in LPMC. A weak absorption peak of benzene
ring C-H bending vibration at around 830 cm−1 could be ob-
served in LPMC and lignin, indicating the existence of lignin
in the capsule wall of LPMC. In addition, the absorption peak
of LPMC at 1050 cm−1, which was ascribed to the stretching
vibration of C-O in urethane bonds, was significantly stronger
than that of PMC due to the reaction between lignin and IPDI
to form lignin-polyurethane.
Encapsulation ratio and drug loading
The encapsulation ratio and drug loading of AVM@LPMC
with different lignin loading were measured. It could be seen
from Table 2 that the higher the lignin loading, the lower the
Table 2 AVM loading and encapsulation ratio of AVM@LPMC with
different amount of lignin on capsule wall
Lignin loading (g) Encapsulation ratio (%) Drug loading (%)
0.61 85.4 4.7
0.73 83.1 4.6
0.84 75.8 4.4
Colloid Polym Sci
encapsulation ratio and drug loading of the microcapsule for
avermectin. Theoretically, if all emulsion droplets obtained
after emulsification could form microcapsules, the AVM
added to the reaction system can be completely embedded
and the encapsulation ratio can reach 100%. However, there
was demulsification during the formation of microcapsules,
and the toluene released from demulsification would volatil-
ize, causing the precipitation of AVM. Therefore, the larger
amount of lignin was added, the more HCl was needed for the
cross-linking of lignin and thus the larger amount of
ethylenediamine hydrochloride was generated in the system,
which would cause more severe demulsification, and thus the
encapsulation ratio of microcapsules would also decrease.
AVM loading refers to the mass percentage of AVM in the
dry microcapsule, which was composed of AVM, toluene, and
wall materials. The larger amount of lignin in wall materials,
the greater the mass of wall material. Therefore, as shown in
Table 2, the drug loading of microcapsules decreased with the
increase of lignin loading.
Sustained release behaviors of microcapsules
The sustained release performance of microcapsules refers to
the continuous release of the encapsulated substances into the
environment over a period of time. As for pesticide microcap-
sules, the better the slow-release performance, the longer the
efficacy lasted. In this paper, ethanol/water (80/20, v/v) mixed
solution was used as the release medium to measure the slow
release performance of AVM@LPMC with different lignin
loading, compared with AVM@PMC and pure AVM (AVM
dissolved in toluene solution).
Figure 10 shows the AVM release kinetics curves of the
microcapsules. It could be seen that the unencapsulated AVM
releases quickly and almost all the AVM released in about
40 h. While the release amount of AVM@PMC only reached
50% after 80 h because the polyurea wall material was too
Fig. 10 Sustained release curves of microcapsules with different lignin
loading
Colloid Polym Sci
dense, which negatively affected the release of AVM and the
pesticide efficacy. The release rate of AVM@LPMC was
slightly faster than that of AVM@PMC, and as the lignin
content in microcapsules increased, the release of
AVM@LPMC was accelerated.
The introduction of lignin could accelerate the release of
AVM from microcapsules, which was mainly because of its
influence on the thickness of polyurea capsule wall. Lignin
which was adsorbed and subsequently deposited on the sur-
face of emulsion droplets would react with IPDI in oil phase to
form lignin-polyurethane, but this lignin layer was relatively
loose and the encapsulation ability of internal AVM was not
strong. Therefore, if the input amount of lignin was larger,
more IPDI would be consumed, resulting in thinner polyurea
layer generated by the reaction between IPDI and water or
ethylenediamine and leading to the faster release of AVM
from microcapsules. That is to say, the slow-release perfor-
mance of the microcapsule mainly came from the relatively
dense polyurea layer inside, and the introduction of different
amounts of lignin could regulate the release rate of the micro-
capsule by changing the polyurea-layer thickness.
Photodegradation studies of microcapsules
Thanks to the polyphenol structure, lignin is endowed with
excellent ultraviolet absorption properties, which means that
LPMC may have anti-photodegradation performance and pro-
tect the encapsulated photolytic materials, such as AVM. The
photodegradation kinetics curves of AVM@LPMC with dif-
ferent lignin loading, AVM@PMC and pure AVM under ul-
traviolet light were measured and the results were shown in
Fig. 11. The photodegradation ratio of AVM of the five sam-
ples was approximately proportional to the time, which is in
line with the first-order kinetic relationship. From the slope of
the curve, it could be detected that the degradation rate of pure
AVM was the fastest with only 20% retention ratio of AVM
after 96 h of ultraviolet irradiation. Although AVM@PMC
Fig. 11 Anti-photolysis performances of microcapsules with different
lignin loading
had a certain anti-ultraviolet effect by encapsulating AVM
inside microcapsules, its degradation ratio was still fast with
only 40% retention ratio of AVM after 96 h. Inspiringly,
AVM@LPMC has the slowest photodegradation ratio, with
a retention ratio of nearly 80% after 96 h, indicating that the
anti-photodegradation effect of AVM@LPMC was signifi-
cantly better than that of AVM@PMC. In addition, the
photodegradation kinetics curves of AVM@LPMC prepared
under three different lignin dosages showed almost the same
trend, indicating that the increased lignin dosages could not
further improve the anti-photodegradation performance of mi-
crocapsules. That was because, the AVM was easy to be re-
leased to the outside of the microcapsules due to the thinning
polyurea layer and could not be protected by lignin when the
amount of lignin was increased, although the lignin layer of
the microcapsule could absorb more ultraviolet rays and en-
hanced the protection of AVM inside the microcapsule.
Conclusion
AVM@LPMC was prepared in this paper, and the single-
factor method was used to study the preparation process.
The results showed that the optimal stirring speed was
200 rpm, and in the case of fixed oil-water ratio (1:9) and oil
phase composition, the optimal ratio of HCl, SDS, and lignin
in water phase is HCl (mmol):SDS (mmol):AL (g) =
0.375:1.25:1. The encapsulation ratio of avermectin, a readily
photolytic pesticide, in microcapsule was up to 85.4%, and the
drug loading was up to 4.7%. AVM@LPMC exhibited good
sustained release performance and the release rate was slightly
faster than polyurea microcapsule. Meanwhile, the larger
amount of lignin loading, the faster release of AVM from
microcapsules. The sustained release ability of the LPMC
was due to the relatively dense polyurea capsule wall. By
combining different amounts of lignin with polyurea, the ad-
justable release rate of microcapsules with composite wall
material could be obtained. Moreover, lignin exhibited ex-
tremely excellent anti-photolysis performance, which could
effectively reduce the degradation of readily photolytic
substances.
Acknowledgments This study is financially supported by the National
Natural Science Foundation of China (21878112, 21676109), Science
and Technology Program of Guangdong (2017B090903003), National
Natural Science Foundation of Guangdong (2017A030308012), and
Guangdong Provincial Special Fund for Modern Agriculture Industry
Technology Innovation Teams (2019KJ140).
Compliance with ethical standards
Conflict of interest The authors declare that they have no conflict
of interest.

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