Progress in Organic Coatings 107 (2017) 43–47

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Progress in Organic Coatings

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

Polyester amide based polyurethane coatings from algae oil and their
larvicidal, anti-ant properties
Aparna Kadam a , Manisha Pawar a , Viresh Thamke b , Omprakash Yemul a,∗
a
School of Chemical Sciences, Swami Ramanand Teerth Marathawada University, Nanded, 431606 Maharashtra, India
b
Department of Chemistry, Savitribai Phule Pune University, Pune 411007, Maharashtra, India

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

Article history: Larvicides play a vital role in controlling mosquitoes in their breeding sites. The present study was
Received 9 November 2016 carried out to establish the larvicidal and anti-ant activities of polyester amide based polyurethane
Received in revised form 10 March 2017 coating from algae oil against mosquitos’ larvae and red ants responsible for diseases of public health
Accepted 10 March 2017
importance. The polyester amide synthesized by N, N-bis (2-hydroxy ethyl) algae oil fatty amide and
ethylene diamine tetra acetic acid through condensation polymerization. After that resin cured with MDI
Keywords:
to obtained polyurethane coatings. Coating performance was tested. Larval mortality of mosquito’s larvae
Polyurethane coating
was observed at 12, 24 and 48 h of exposure to different weight of PU films. The efficacy of polyurethane
Larvicidal
Anti-ant property
coating films against mosquitos’ larvae (LC50 and LC90 ; 5.35 and 17.15 mg/mL) respectively were observed
Fatty amide by probit analysis while the mortality of ants also observed at 2, 4, 6 and 12 h of exposure to PU coatings.
Polyester amide Polyurethane coatings can be recommended effectively in mosquitos and ants controlling practices. The
cytocompatibility study was also carried out and study shows good results.
© 2017 Elsevier B.V. All rights reserved.

1. Introduction able properties against such vectors. Polymers a modern material

The mosquito is the major vector of dengue and malaria disease.
The continuous use of various kinds of insecticides has increased
mosquitoes’ resistance [1]. This action causes low insecticidal
susceptibility in the mosquitoes and thus contributes to further
development of their population. Ants are not much harmful to
public health but they cause nuisance as they enter homes in search
of food. Currently mosquitos and ants controlling practices are
mainly dependent on synthetic chemical substances. The use of
synthetic insecticides to control insect pests leads to deterioration
of ecosystem suggesting the need for other alternatives [2]. Plant
derived natural products have the advantage over synthetic insec-
ticides of being harmless to non-target organisms and have been
used by human communities as mosquito larvicides, insect growth
regulators, repellents and ovipositional attractant [2].
Therefore, to find out effective and economically viable ways to
control mosquitos and ants, there is need to search for such lar-
vicidal and anti-ant active compounds which are environmentally
safe, cost effective, biodegradable, new methods consisting of legal
scientific and technological weapons with more powerful combat-
∗ Corresponding author at: School of Chemical Sciences, Swami Ramanand Teerth
Marathwada University, Nanded − 431 606, India.
E-mail address: omyemul@gmail.com (O. Yemul).
has replaced the traditional materials like glass, ceramic, metal,
leather, wood, stone, horn, bone and paper. It has been possible
due to its extraordinary properties. Polymers are widely used for
many different tools, such as catheters, surgery plaques and screws,
as well as scaffolds for tissue engineering or drug delivery [3]. How-
ever, even though polymers have already brought in medicine (e.g.,
surgery, implants, delivery systems), allowing incredible benefits
for the patient, rather simple polymer structures have been used
and developed so far. It is possible due to their biological proper-
ties such as biocompatibility, biodegradability, antimicrobial etc.
Therefore it is important to study the biological activity of polymer
for their application in related field.
So the aim of our study is to investigate larvicidal and anti-ant
properties of algae oil based polyurethane coatings. Considering
these facts in the present work, initially algae oil fatty amide (AOFA)
and algae oil polyester amide (AO-PEA) have been synthesized. AO-
PEA resin obtained by the modification of AOFA with EDTA looks
to an attractive proposition. Poly (urethane-fatty amide) (AO-UFA)
and poly (urethane-ester amide) (AO-UPEA) have been prepared
from AOFA and AO-PEA by curing with MDI respectively. Further we
studied the larvicidal and anti-ant properties of these polyurethane
coatings.

http://dx.doi.org/10.1016/j.porgcoat.2017.03.013
0300-9440/© 2017 Elsevier B.V. All rights reserved.
44 A. Kadam et al. / Progress in Organic Coatings 107 (2017) 43–47
2. Materials and methods
All chemicals/reagents used in this experiment were of lab-
oratory grade and were procured from either SD Fine Chem or
Spectrochem, India. They were used without further purification.
Algae oil from chlorella microalgae (IV 120 and acid value 0.2 mg
KOH/g) was a gift sample from Soley Institute Turkey. Actively
swimming larvae were obtained from the tank of school of Life
sciences, SRTM University, Nanded (MS). Red ants were collected
from the moistened area of the house.
2.1. Synthesis of Algae oil based fatty amide (AOFA)
AOFA was synthesized as per the procedure previously reported
in literature for jatropha oil and Mahua oil [4,5].
2.2. Synthesis of Algae oil based polyester amide (AO-PEA)
AO-PEA was synthesized as per the procedure previously
reported in literature for linseed oil [6].
2.3. Preparation of algae oil poly (urethane −fatty amide)
(AO-UFA) and poly (urethane-ester amide) (AO-UPEA) coatings
AO-UFA and AO-UPEA coatings prepared as per the procedure
previously reported in literature by Gaikwad et al. [7].
2.4. Larvicidal assay
Larvicidal bioassay method, proposed by Nour et al. [8] was con-
ducted with slight modification. A number of 10 actively swimming
larvae (2nd and 3rd instars) were released into 50-mL capacity
beakers having 20 ml water of each set. Untreated set of larvae in
(tap) water as Control (Set a) was run for comparison while set
b, c, d and e contained 0.1 g of AO-UFA 1, AO-UFA 2, AO-UPEA 1
and AO-UPEA 2 respectively. For feeding of larvae there was pro-
vided equal amount of starch. The beakers were stored at room
temperature at about 29 ◦ C ± 2 ◦ C. Mortality of larvae was recorded
after 1-, 3-, 6-, 24- and 48-h of exposure and moribund larvae were
counted as dead. Toxicity and activity, were reported as LC50 and
LC90 , representing the concentrations in ppm that killed 50 and
90%, respectively of larvae in 24 and 48 h. While the same set of
larvae experiments have been performed for 0.3 g, 0.5 g of PU films.
2.5. Anti-ants assay
Anti-ants bioassay, proposed by Mehmood et al. [9] was per-
formed with minor changes. A number of 10 healthy ants were
released into plastic chamber coated with PU coating and provided
with aeration. 4 sets were prepared Set A coated with AO-UFA 1,
Set B coated with AO-UFA 2, Set C coated with AO-UPEA 1 and
Set D coated with AO-UPEA 2. For feeding of larvae there was pro-
vided equal amount of sugar in each set. The chambers were stored
at room temperature at about 29 ◦ C ± 2 ◦ C. Mortality of ants was
recorded after 1-, 2-, 4-, 6-, 8-, 12- and 24-h of exposure and mori-
bund ants were counted as dead.
2.6. Cytocompatibility studies
Pieces of polymers (AO-UFA 2 and AO-UPEA 2) having a diameter
of 15 mm were surface sterilized by immersing in 70% ethanol for
1 h and later exposed to UV radiation for 1 h. The polymer pieces
were then placed into the bottom of the well in a 24 well plate.
The tissue culture medium used contained 2% FBS, 1% Antibiotic
solution along with DMEM for PANC-1cells and MEM for BS-C-1
cells. Around 5 × 104 cells were seeded on polymer placed well
along with the media and kept in the CO2 incubator at 37 ◦ C and
5% CO2 for 48 h. After incubation, medium was removed and poly-
mers in the wells were washed twice with 1X Phosphate buffer
saline. The 40 ␮g/well MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-
diphenyltetrazolium bromide) was added in each well and the plate
was kept for incubation for 4 h at 37 ◦ C. MTT was removed and
300 ␮l DMSO was added. Now 300 ␮l solution of all wells from 24
well plate were distributed in triplicates in a 96 well tissue cul-
ture plate and colour of formazan formed was measured at 570 nm
in multimode plate reader (PerkinElmer) [10]. The cells grown in
absence of polymer were considered as negative control.
3. Characterization
Statistical analysis of the experimental data was performed
using computer software Excel 2007 to find the lethal concentra-
tions required to kill 50% (LC50 ) and 90% (LC90 ) of larvae in 48 h
by probit analysis with a reliability interval to establish regression
equations (Y = mortality; X = concentrations) and regression coeffi-
cient values.
PANC-1(human pancreatic cancerous cell line) and BS-C-1
(African Green monkey kidney cell line) cell lines were purchased
from National Centre For Cell Science (NCCS, India) and cultured in
25 cm2 tissue culture flask containing Dulbecco’s Modified Eagle’s
Medium (DMEM, Gibco, USA) and Modified Eagle’s Medium (MEM,
Gibco, USA) supplemented with 10% Fetal Bovine Serum (FBS,
Gibco, USA). The 5% CO2 and 37 ◦ C temperatures was maintained
in humidified atmosphere.
4. Results and discussion
Polymers have many applications in medicine (e.g., surgery,
implants, and delivery systems) due to their biological properties.
As we know, mosquitos are the cause of many diseases and if ants
enter in home they cause nuisance. In order to find out the effective
way for this, in the present work, we studied the larvicidal, Anti-ant
properties of algae oil based PU coatings.
The algae oil (non-edible plant oil) chemical composi-
tion, renewable nature has motivated researchers to develop
polyurethane coatings and studied their larvicidal, Anti-ant prop-
erties from it. The fatty acid composition of algae oil was estimated
by esterification of acids using methanol followed by GC analysis
and are given in Supplementary File.
4.1. Coating properties
PU coatings were applied on MS panels and on particle boards
too. The coating performance was assessed by studying parameters
such as drying time, gloss, scratch hardness, mar resistance and
cross cut adhesion etc. The tested values are listed in Table 1. To
determine the optimum composition of the coating that provided
the best performance in case of AO-UFA and AO-UPEA coatings, two
different compositions were prepared with two different loadings
of MDI (1.0–1.1 mol). The properties of compositions with 1.1 mol
of MDI are found better than 1.0 mol of MDI. Good impact resistance
shows higher adhesion of coatings with the substrate through coor-
dinate linkage with nitrogen and oxygen atoms. Excellent flexibility
in the coatings due to the presence of long aliphatic chain and tetra
functional EDTA moieties is found. The gloss at 60◦ is found to be
between 55 and 59. It could be inferred that the gloss of the resins
is independent of the amount of MDI used.
 A. Kadam et al. / Progress in Organic Coatings 107 (2017) 43–47 45

Table 1
Coating properties of AO-UFA and AO-UPEA coatings.

Resin code Drying time Gloss at 60◦ Scratch Mar Pencil hardness B Flexibility 1/800 Cross cut
(min) hardness (Kg) resistance (g) and H Mandrel adhesion (%)

AO-UFA 1a 27 55 1.8 110 2H PASS 94

AO-UFA 2b 25 59 1.9 120 2H PASS 95
AO-UPEA 1c 27 55 1.8 110 2H PASS 94
AO-UPEA 2d 25 59 1.9 120 2H PASS 95
a
AO-UFA 1: NCO/OH ratio 1:1.
b
AO-UFA 2: NCO/OH ratio 1.1:1.
c
AO-UPEA: NCO/OH ratio 1:1.
d
AO-UPEA: NCO/OH ratio 1.1:1.

4.2. Larvicidal activity of polyurethane films
The aim of this study is to investigate the larvicidal property
against mosquito larvae. The larvicidal activity images of different
combinations of PU films are shown in Fig. 1A and larvicidal activity
graphs of PU films for statistical study are exhibited in Fig. 1B.
The statistical study of different weights of PU films against
mosquito larvae compared to that of the control is shown in Table 2
varied with different weights and exposure time. The results of
regression analysis revealed that the mortality rate (Y) was pos-
itively co-related with the period of exposure(X) with regression
co-efficient close to 1. The result of log-probit analysis revealed that
LC50 values gradually decreased with the exposure period with the
lowest value at 48 h of exposure (Table 2). The study indicates that
the most effective coating film is that which have low LC50 value
and show highest mortality. The data reported that after 24 h of
mosquito larvae exposure, the highest mortality recorded for AO-
UPEA 2 with LC50 and LC90 values of 5.35 and 17.15 mg/mL while
LC50 and LC90 values of other PU films are respectively shown in
Table 2.
Mosquito control at the larval stage is effective method because
they are localized in space and time resulting in less-dangerous
out comes while the fight against adult is temporary and unsat-
isfactory. Long term extensive application of synthetic insecticides
results to the accumulated side effects of toxicant and environmen-
tal contamination. Polyurethane coating have promising larvicidal
efficacies may be due to the starting material used algae oil [11],
EDTA [12] and urethanes over synthetic insecticides as it is par-
tially biobased and less toxic to environment. Additional advantage
is the use of coating that is to coat the walls of houses, windows,
water hold containers in order to kill adult mosquitoes and their
larvae. However further studies are needed to investigate the chem-
ical structure and mechanism which are responsible for larvicidal
activity.
4.3. Anti-ants activity of polyurethane coating
Ants are not much harmful to public health but they cause nui-
sance as they enter homes in search of food. These ants require
moisture to survive. Increasing demand for natural sources of insec-
ticides encouraged us to undertake a comprehensive study of the
insecticidal and repellent activities of the PU coatings against red
ants normally found in houses. They can infest buildings, partic-
ularly foundation forms in contact with soil, becoming a minor
nuisance.
The study is conducted to evaluate the insecticidal activity of
PU coatings obtained from algae oil against red ants. The repellent
effect revealed that the AO-UPEA 1, 2 took least time, i.e. 2.5 h for
100% repellency whereas AO-UFA 1, 2 took 4 h with respect to AO-
UPEA 1, 2. The anti-ants activity of PU films are represented in Fig. 2.
The main benefit of polyurethane coating against anti-ant activity
is that we can coat the area where the ants cause nuisance as they
enter homes in search of food. However further studies are needed

Fig. 1. A) Larvicidal activity images of a) Control, b) AO-UFA 1 c) AO-UFA 2, d) AO-UPEA 1, e) AO-UPEA 2 and B) Larvicidal activity of a) AO-UFA 1, b) AO-UFA 2 c) AO-UPEA
1and d) AO-UPEA 2 polyurethane coating films.
46 A. Kadam et al. / Progress in Organic Coatings 107 (2017) 43–47

Table 2
Statistical study of different weights of polyurethane films against mosquito’s larvae.

Polymer Code % Mortality (hours) R2 Regression equation LC50 (mg/ml) LC90 (mg/ml)
a
AO-UFA 1 12 R2 = 0.852 y = 1.485x + 4.761 30.46 57.40
24 R2 = 0.754 y = 2.171x + 16.19 15.57 33.99
48 R2 = 0.697 y = 2.628x + 23.81 9.96 25.18

AO-UFA 2b 12 R2 = 0.754 y = 2.171x + 16.19 15.57 33.99

24 R2 = 0.697 y = 2.628x+ 23.81 9.96 25.19
48 R2 = 0.637 y = 3.314x + 35.23 4.45 16.52

AO-UPEA 1c 12 R2 = 0.818 y = 3.525x + 20.33 8.41 19.76

24 R2 = 0.457 y = 3.050x + 40.67 3.059 16.17

AO-UPEA 2d 12 R2 = 0.623 y = 3.525x + 20.23 8.41 16.17

24 R2 = 0.818 y = 3.389x + 31.86 5.35 17.15

R2 −regression coefficient values, LC50 and LC90 −lethal concentration that kills 50% and 90% mosquito’s larvae respectively.
a
AO-UFA 1: NCO/OH ratio 1:1.
b
AO-UFA 2: NCO/OH ratio 1.1:1.
c
AO-UPEA 1: NCO/OH ratio 1:1 and.
d
AO-UPEA 2: NCO/OH ratio 1.1:1.

Fig. 2. Anti-ants activity of a) Set A: Control, b) Set B: AO-UFA 1 coated c) AO-UFA 2 coated d) Set C: AO-UPEA 1 coated and d) Set D: AO-UPEA 2 coated PU coatings.

to investigate the chemical structure and mechanism which are

responsible for anti-ant activity.

4.4. Cytocompatibility studies

Biocompatibility has been defined previously as “the ability

of a material to perform, with an appropriate host response in a
specific application”. Biomedical materials have been successfully
developed for in vivo uses and in vitro tissue cultures. Because
biomedical materials are either directly or indirectly in contact with
human organs, tissues, blood or skin, the in vivo stability, toxicity,
biodegradability and biocompatibility of the materials are impor-
tant considerations. With this background we would like to check
the biocompatibility of PU coatings.
The yellow coloured MTT get transformed into violet coloured
formazan crystals with the help of mitochondrial dehydrogenase
enzyme, which is relative to amount of live cells. The formazan
crystals were easily dissolved by the DMSO and intensity of colour
differs with the number of live cells. In the PANC-1 cells incubated Fig. 3. % survival of PANC-1 cells and BSC-1 cells in presence of PU films.
with PU coated paper, it is found that AO-UFA 2 and AO-UPEA 2
showed survival 103% and 120%, which illustrates that the PU coat-
ing AO-UFA 2 and AO-UPEA 2 did not exhibit any toxicity on the [13] evaluated the biocompatibility of composites prepared from
PANC-1 cells and are biocompatible. Results also indicated that AO- chestnut shell fibre (CSF) and poly (hydroxyalkanoate) (PHA), also
UPEA is more biocompatible than AO-UFA on exposure of PANC-1 CSF and glycidyl methacrylate grafted PHA (PHA-g-GMA) on the
cells. human foreskin fibroblasts (FB) cell.% survival rate of PANC-1 cells
However on exposure of BS-C-1 cells to both PU coatings and BSC-1 cells in presence of PU films is shown in Fig. 3.
revealed that in presence of AO-UFA 2 and AO-UPEA 2 only 69%
and 29% survival rate is observed indicating considerable toxicity 5. Photographs of polyurethane coatings
of these PU coating. Results indicated that AO-UFA 2 is more bio-
compatibility than AO-UPEA 2 on exposure of BSC-1 cells. Wu et al. Fig. 4
 A. Kadam et al. / Progress in Organic Coatings 107 (2017) 43–47 47

Fig. 4. Images of A) AO-UFA 2 and B) AO-UPEA 2 PU coatings on steel and particle board.

6. Conclusions Appendix A. Supplementary data

In the present work, algae oil is successfully utilized in the Supplementary data associated with this article can be found,
preparation of polyurethane coatings. PU coatings showed good in the online version, at http://dx.doi.org/10.1016/j.porgcoat.2017.
coating properties. In conclusion, our findings showed that algae 03.013.
oil based polyurethane coating can be developed as ecofriendly lar-
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