Chapter

5 Stable O/W Emulsions of Karanj, Castor and Neem Oil for Pesticide Applications.

Chapter 5 Storage Stable O/W Emulsions of Karanj

(Pongamia glabra), Castor (Ricinus communis
L.) and Neem Oil (Azadirachita indica A. Juss)
for Pesticide Applications

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5.1 Introduction
Synthetic agrochemicals are responsible for ecological imbalance, food chain
disruption, stream water contamination resulting into human and animal toxicity and
traces in agricultural products even in breast milk. These agrochemicals also have preapplication hazards such as process pollution, occupational hazard and handling exposure.
The exaggerated perception of such hazards has forced researchers to find a
comparatively safer alternative for these chemicals. Some of the easily available nonedible vegetable oils are having major anti insect actives in addition to their fatty acid
composition.
Karanj based products are found to be effective against insect pests of stored
grains, field and plantation crops, and household commodities.315 More than nineteen
biologically active components have been identified from karanj plant. Oil, organic leaf
extract, methanolic and aqueous seed extract, of karanj have shown potential to act as
oviposition deterrents, antifeedants, antibacterial, antifungal, mosquito repellent and
larvicidal against a wide range of insects.316 - 321 Karanj oil (Pongamia glabra) contains the
non-glyceride toxins Karanjin and Pongamol.322, 323 The bioefficacy of Karanj, and Neem
oil against Mustard aphid, Lipaphis erysimi (Kaltenbach) under different cropping
systems were shown significant reduction in the mustard aphid population. Effects of
karanj seed extracts on growth and development of Plutella xylostella L. (Lep.,
Yponomeutidae) and on oviposition and egg hatching of Plutella xylostella (Lepidoptera:
Yponomeutidae) have proved their potential as a promising agrochemical.324 - 326
The ricin and ricinine are active ingredients of Ricinus communis that acts against
S. frugiperda. Castor seed extract keeps more insecticidal and insectistatic potential than
the leaf extract.327 It resulted in effective seed protection from Z. subfasciatus infestations
comparable to the control of malathion with least seed damage, weight loss and without
any adverse effect on germination capability of the seeds.328 Castor and Hazelnut oil have
shown insecticidal activity against Callosobruchus maculatus (F.) (Coleoptera:
Bruchidae),329 Musca domestica330 and termites.331
A tetranortriterpenoid Azadirachtin (C35H44O16) present in Neem oil is the most
potent, natural insect feeding deterrent as well as insect growth regulator to which over
200 species of insects are found to be susceptible.332, 333 The activity of Neem seed oil on
various pests is explored viz. Lutzomyia longipalpis (Diptera: Psychodidae),334
Rhipicephalus (Boophilus) microplus,335 whorl larva, stem-borer and panicle insect pests
of

sorghum,336

red

flour

beetle,

Tribolium

castaneum (Herbst)

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Tenebrionidae), lphitobius diaperinus (Coleoptera: Tenebrionidae).337

In addition to

synergistic fungicidal activity Neem oil is responsible for repellency against Phlebotomus
orientalis, P. bergeroti (Diptera: Psychodidae),338 large pine weevil, Hylobius abietis L.
(Coleptera, Curculionidae).339,

340

The in vitro toxicity of neem seed oil was proved

against the larvae of a one-host tick, Boophilus decoloratus (family: Ixodidae or hard
tick,commonly known as blue tick) and for ovicidal activity on the eggs of bhendi.341
Tropical environment is suitable for Karanj, Castor and Neem tree. They are
grown in a wide area stretching from Australia, India to south and North American
territory. These oils are readily available in market at reasonable cost. Profitable oil
isolation methods from well dried seeds include processes like cold dry pressing, steam
distillation and solvent extraction technique. Formulation of crude vegetable oil is
advantageous over the isolation and formulation of actives as it includes other potentially
biotoxins present in seed and possesses excellent storage stability.342, 343
Vegetable oils, like all other botanical resources, will considerably vary with
respect to their bioactive content depending upon the variation in tropical conditions and
efficiency of oil extraction technique. The agrochemical and medicinal importance of
Karanj, Castor and Neem oil is proved by the series of above discussed work. However,
direct application of vegetable oil has drawback like high viscosity, less spreadability,
cost of oil, probable overdose resulting into unconsumed residues in soil and aquatic
system. This demands a suitable economically feasible formulation technique for their
widespread agrochemical applications. The present study was focused on developing the
stable O/W emulsions with varying oil content while keeping emulsifier content to
minimum level. Emulsifying agents are well known to play an important role in the
stability of emulsion and proper selection of the emulsifying agents gives the synergistic
effect.344 The blend of nonionic surfactants viz. NP-13 (HLB No. 15) and Span-80 (HLB
No. 4.5) was used for emulsion formation. The emulsion stability studies were carried out
at 30C 0.5C for 24 hrs and the stable emulsions were further tested for their
rheological and insecticidal properties.

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5.2 Materials and Methods

5.2.1 Materials
Karanj, Castor and Neem oil were procured from M/s. Ashwin Pharma Ltd.
Mumbai, India. NP13 (Nonyl Phenol Ethoxylate 13 Moles) and Span80 (Sorbitan
Monooleate) were procured from M/s. SD Fine Chemicals, Mumbai, India. All other
chemicals were of analytical grade. Deionized water was used for experimental work.
Mosquito larves (Third stage/early fourth stage) were supplied by Dr. Balasaheb Sawant
Konkan Krishi Vidyapeeth. (Dr. Balasaheb Sawant Konkan Agriculatural University,
Dapoli, India). All other chemicals were obtained from M/s. S. D. Fine Chemicals Ltd.
Mumbai, India and used without further purification.
5.2.2 Methods
5.2.2.1 Preparation of Emulsion
Emulsions were prepared under the influence of external agitation with the help of
homogenizer. In each case calculated quantity of emulsifier blend was dissolved in oil
phase which was then added to the aqueous phase under the influence of external
agitation with the help of electrically driven motor (2000 RPM- Remi Motors Ltd. India).
The prepared emulsions were then observed for macroscopic stability for 24 h at 30C
0.5C. The emulsion instability was expressed in terms of creaming while the white
milky layer is treated as a stable emulsion.
5.2.2.2 Effect of Various Parameters
Initially the emulsifier content was kept constant at 3% (w/w) level and oil content
was varied from 1% (w/w) to 15% (w/w). However, the emulsions were found to be
unstable. This procedure was repeated by varying the emulsifier content. Emulsifier blend
of various HLB numbers were prepared by properly adjusting the percentage of
individual surfactant as shown in Table 5.1.

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Table 5.1 - Preparation of emulsifier blends with varying HLB number.

NP-13 (wt. %)
Span-80 (wt. %)
HLB number of emulsifier blend
0.00
100.00
4.5
4.60
95.40
5
14.28
85.70
6
23.80
76.20
7
33.30
66.70
8
42.86
57.10
9
52.40
47.60
10
61.90
38.10
11
71.40
28.60
12
81.00
19.00
13
90.50
9.50
14
100.00
0.00
15
5.2.2.3 Rheological Characterization
Rheological properties of the final emulsion intended to be used as a sprayable
product decides the physical stability, ease of spraying, spreadability and its overall
performance.345 In case of O/W emulsions viscosity is typically governed by the amount
of internal oil phase. Haake VT500 Viscometer (Germany) operated by Thermo
Rheowin 2.97 software equipped with DC-5 cooling system was used for rheological
characterization. The 35 g of each emulsion sample is subjected to a shear programme
from 6.45S-1 to 645S-1 with 1 minute time interval. Each rheological measurement was
performed in three replications and average S.D. values were plotted. Rheological
parameters like consistency index (K), flow index (n) were calculated by applying
Power law model.346, 347
5.2.2.4 Spreadability on plant leaves and average globule size
The emulsions were checked for there spreadabilty on horizontal coconut leaves.
A circle of 1 cm diameter was drawn on leaf and a drop of emulsion was placed at the
centre of the circle. The average time required for emulsion drop to touch the opposite
ends of the circle is noted for three independent replications. The average globule size of
emulsion is determined in five replications using COULTER LS230 instrument.
5.2.2.5 Larvicidal Activity
The most efficient and simpler way to control mosquito population is
larviciding.348 In the present paper the larvicidal activity of developed formulations were
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tested using twenty five larve of each mosquito species at 25-28C with a photo period of
12 hr L: 12 hr D.349 Initially a series of trial experiment (non replicated) were conducted
with different test formulations to optimize the dose with a geometric factor of 2.0 giving
595% mortality. Test formulations were composed of single as well as equal volume
mixtures of two and more emulsions. Single oil emulsion concentrations used were 0.01,
0.02, 0.04, 0.08, 0.16, 0.32, 0.64, 1.28, 2.56 and 5.12% whereas for an equal volume
mixture of two and three oil emulsions the lower limit of the test formulation percentage
were started from 0.0025 and 0.001 respectively. Each measurement is performed in four
replications. After 24 hr, the number of dead larves was counted and the data was
subjected to probit analysis350 using NCSS 2007 statistical package. Increase in mosquito
larvicidal efficacy is expressed in terms of fold increase and calculated using the formula:
Fold increase = LC50 or LC90 or LC95 of individual emulsion treatment / LC50 or
LC90 or LC95 of combination emulsion treatment.

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5.3 Results and Discussion

5.3.1 Optimum Emulsion Parameters
A series of stable emulsion products were developed by trial and error method
considering the ease of emulsion formation and emulsion stability for 24 hrs at 30 0.5C
are depicted in Table 5.2. Practically the apparent viscosity, specific gravity of these oils
was matching and the peak emulsification performance is achieved by matching HLB
numbers. The equipment used for emulsification such as beaker dimensions, speed of
agitation and agitation blade geometry were found to be ideal in case of emulsions with
10% oil concentration as the same quantity of emulsifier i.e. 5% was required. The
increase in emulsion particle size is obtained with increasing internal oil phase. The
increase in oil content has increased viscosity of the emulsion there by reducing
spreadability of emulsion droplet.
Table 5.2 - Optimum emulsion parameters.
For
mula

Oil

Composition (wt %)

Oil
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24

Karanj

Castor

Neem

5
10
15
20
25
30
35
40
5
10
15
20
25
30
35
40
5
10
15
20
25
30
35
40

Emulsifier
blend
2.5
5.0
8.3
11.2
12.5
13.7
15.2
17.3
2.4
5.0
7.8
10.3
11.9
13.5
14.6
16.1
3.1
5.0
7.9
10.1
12
13.5
14.3
15.9

NP
13:
Span 80

Phase
mass ratio
(m)

Avg.
Spred
ability
(S)

Avg. globule
size (m)

3.3:6.7

0.0540
0.1176
0.1955
0.2906
0.4000
0.5328
0.7028
0.9367
0.0539
0.1176
0.1943
0.2869
0.3961
0.5309
0.6944
0.9111
0.0544
0.1176
0.1945
0.2861
0.3968
0.5309
0.6903
0.9070

3
7
9
12
19
27
38
42
2
5
9
14
20
24
26
34
2
4
7
12
16
23
30
37

0.972
1.710
2.073
4.085
6.097
9.452
12.983
17.35
0.345
0.723
1.570
2.654
3.871
6.286
8.592
13.233
1.004
2.320
3.002
4.975
6.087
9.073
11.238
15.985

Water
92.5
85
76.7
68.8
62.5
56.3
49.8
42.7
92.6
85.0
77.2
69.7
63.1
56.5
50.4
43.9
91.8
85.0
77.1
69.9
63
56.5
50.7
44.1

9.05:0.9
5

8.1:1.9

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5.3.2 Effect of Oil Content on Stability

At a given emulsifier percentage (3% w/w) as the oil percentage increased, the
stability of emulsion increased up to a maximum value at 10% of oil content and further
increase in the oil content was resulted into the destabilization of emulsion. Emulsion
instability at lower oil content may be due to the difference in the gravitational forces
whereas the surfactant deficiency unable to form the tight monolayer around dispersed oil
droplet caused instability at higher oil content. This ultimately resulted into successive
collisions of oil droplets leading to coalescence and destabilization of emulsion. Figure
5.1 shows the effect of oil content on stability of emulsion.

Figure 5.1 - Effect of oil content on stability of emulsion at 3% (w/w) emulsifier

content.

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Chapter 5 Stable O/W Emulsions of Karanj, Castor and Neem Oil for Pesticide Applications.

5.3.3 Effect of Emulsifier Content on Stability

Polyethoxylated nonionics are soluble in water due to hydrogen bonding of water
molecule to the oxygen atom of polyoxyethylene chain. The surfactant molecules from oil
phase and water phase get associated in a particular way at the O/W interface. Sufficient
emulsifier concentration is necessary to produce the tightly packed monolayer of
surfactant molecules at the interface which acts as a mechanical barrier to flocculation
and coalescence of oil droplets thereby stabilizing the emulsion. The effect of emulsifier
content on the stability of emulsion is shown in Figure 5.2

Figure 5.2 - Effect of emulsifier content on stability of emulsion at 10% (w/w) oil
content.
At emulsifier concentration of 5% the maximum stable emulsion was obtained.
The emulsifier concentration lower than this was unable to prevent the flocculation and
coalescence of oil droplets and consequently destabilized the emulsion. The emulsifier
concentration above 5% also destabilized the emulsion due to the temporary emulsifier
saturation in inter droplet region.

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5.3.4 Effect of HLB Number of Surfactant Blend on Stability

Maximum stability for karanj, castor and neem oil emulsions is obtained at HLB
of surfactant blend 8, 14 and 13 respectively. The HLB numbers of surfactant blend fairly
match with those of oils, which is one of the conditions to get the stable emulsion. Figure
5.3 shows the effect of HLB number of surfactant blend on the stability of emulsion.

Figure 5.3 - Effect of HLB of surfactant blend on stability of emulsion at 10% (w/w)
oil content and 5 % (w/w) emulsifier content.

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Chapter 5 Stable O/W Emulsions of Karanj, Castor and Neem Oil for Pesticide Applications.

5.3.5 Effect of Hardness of Water on Stability

In agricultural applications the hardness of water is the matter of great concern as
it varies with the location. Figure 5.4 shows the effect of hardness of water on stability of
emulsion. The maximum limit of hardness of water was found to be 342 ppm, above
which the emulsions were gradually destabilized. Neem oil emulsion showed maximum
tolerance for hard water followed by karanj oil emulsion and castor oil emulsion showed
the least tolerance.

Figure 5.4 - Effect of hardness of water on stability of emulsion at 10% (w/w) oil
content and 5% (w/w) emulsifier content.

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Chapter 5 Stable O/W Emulsions of Karanj, Castor and Neem Oil for Pesticide Applications.

5.3.6 Rheological Characterization

All formulated emulsions showed a pseudoplastic flow behavior i.e. shear
thinning nature which is the characteristics of a good sprayable O/W emulsion product.
Viscosity measurement of karanj oil emulsion is shown in Figure 5.5.

Figure 5.5 - Viscosity measurement of formulated karanj oil emulsions.

The oil content is the main viscosity determining parameter being viscous and
variable. In case of all emulsion varying oil content a rapid decrease in viscosity at the
initial shear rates is obtained which further decreased slowly at higher shear rates which
an indication of increased spray efficiency. Rheogram of karanj oil emulsion is shown in
Figure 5.6. Rheograms of castor and Neem oil emulsion are found to be identical in graph
nature and flow behavior hence only rheological parameters are given for them.

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Figure 5.6 - Rheogram of formulated emulsions.

Table 5.3 contains the values for power law model showing the empirical
consistency (k) and flow behavior indices (n). The correlation coefficient (r2) values
ranging from 0.989 to 0.998 suitably explains the experimental data. The increase in oil
percentage has resulted into increase in apparent viscosity thus statistical increase in
empirical consistency index which is the indication of viscosity.

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Chapter 5 Stable O/W Emulsions of Karanj, Castor and Neem Oil for Pesticide Applications.

Table 5.3 - Rheological parameters of stable emulsions.

Formula

Consistency Index (k)

Flow Index (n)

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24

62 3.4
70 8.1
76 1.5
83 12
90 5.3
113 16.2
125 10.5
130 2.5
57 6.0
60 8.5
72 7.1
85 12.0
95 10.4
110 22.3
129 19.0
136 23.3
47 3.2
53 5.5
67 6.1
79 7.2
92 10.2
123 15.6
136 20.2
157 23.5

0.34 8.5 10-3

0.35 1.5 10-3
0.46 5.6 10-3
0.46 7.4 10-2
0.34 3.9 10-3
0.34 5.7 10-2
0.47 2.9 10-2
0.46 2.1 10-2
0.34 8.9 10-3
0.34 2.2 10-2
0.47 3.2 10-2
0.46 8.2 10-2
0.34 2.3 10-2
0.34 5.1 10-3
0.48 6.5 10-3
0.47 1.2 10-2
0.44 4.2 10-2
0.37 2.9 10-2
0.46 3.1 10-2
0.39 7.1 10-3
0.42 4.2 10-2
0.45 3.4 10-2
0.34 6.9 10-2
0.34 2.5 10-2

Correlation coefficient
(r2)
0.991
0.994
0.994
0.992
0.992
0.993
0.994
0.991
0.989
0.992
0.993
0.993
0.991
0.993
0.989
0.994
0.989
0.924
0.990
0.994
0.998
0.992
0.993
0.989

Values of flow index were consistently below unity which indicates a

pseudoplastic flow behavior. The consistency index increased with increase in oil content.
From the rheological data the formulated emulsions were found to have optimal viscosity,
pseudoplastic flow nature and good consistency.
5.3.7 Larvicidal Activity:
The results of the larvicidal activity are depicted in Table 5.4 to 5.7. From the
study, it was clearly observed that the combination of Karanj, Neem and Castor oil
emulsions were found to be more effective than their individual treatment against all the
mosquito larvae tested. The increase in efficacy of the combination Karanj, neem &
castor oil emulsion treatment (1:1:1) over individuals in all the mosquito larvae tested
was found to range from 8.4 to 9.5 fold for karanj oil, 5.5 16.0 fold for neem oil and
16.2 to 23.9 fold for castor oil emulsion in terms of LC50. Similar increaments were also

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Chapter 5 Stable O/W Emulsions of Karanj, Castor and Neem Oil for Pesticide Applications.

observed in case of LC90 and LC95 for a combination treatment of karanj, neem and castor
oil emulsion (1:1:1)
The overall order of larvicidal activity is found to be,
karanj, neem & castor (1:1:1)>karanj and neem (1:1) > Neem and castor (1:1)> Karanj
and neem (1:1)> karanj > Neem > castor
Hence, the oil formulations are economically and practically feasible method to
control mosquito larvae. From the present investigation, it is obvious that the performance
of combined application of neem and karanj oil emulsion was better against the mosquito
larvae than their individual application. Though the individuals are good against mosquito
larvae, the synergistic effect is well exhibited in this experiment. Use of the above
products in alternation with the chemical insecticides may help in causing delay in the
development of resistance in variety of pests.

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Table 5.4 - Larval mortality data of karanj, neem and castor oil emulsions.
Emulsion
Corrected larval mortality at 24 hr (%)*
concentrations (%)
Culex
Aedes aegypti Anopheles stephensi
quinquefasciatus
Karanj oil
0.01
5 (0.35)
6(1.91)
8(1.2)
0.02
14 (2.95)
13(2.58)
20(3.4)
0.04
36 (3.2)
29(1.63)
52(4.2)
0.08
42 (1.95)
37(3.65)
59(2.83)
0.16
63 (2.5)
68(3.27)
65(2.75)
0.32
74 (3.7)
78(2.52)
78(3.52)
0.64
84 (2.58)
86(2.58)
89(3.80)
1.28
91 (3.27)
92(1.23)
94(2.38)
2.56
100
100
100
Untreated control
0.00
0.00
0.00
Castor oil
0.01
0.00
0.00
0.00
0.02
4(0.48)
7(0.45)
2(0.85)
0.04
12(0.93)
14(1.03)
15(0.85)
0.08
32(1.28)
25(0.64)
27(1.16)
0.16
48(1.64)
47(0.82)
41(2.10)
0.32
65(2.83)
70(1.02)
59(3.64)
0.64
78(3.54)
81(1.85)
75(3.97)
1.28
86(3.73)
92(3.28)
93(4.86)
2.56
93(3.95)
100
98(4.07)
5.12
100
100
Untreated control
0.00
0.00
0.00
Neem oil
0.01
2(0.26)
0.00
0.00
0.02
19(0.83)
9(0.47)
7(0.38)
0.04
37(1.09)
17(0.67)
15(1.16)
0.08
59(1.59)
33(1.28)
28(1.24)
0.16
76(2.06)
69(1.83)
55(2.74)
0.32
92(2.68)
85(2.06)
79(2.93)
0.64
98(3.06)
91(2.72)
85(3.02)
1.28
100
100
93(3.9)
2.56
100
Untreated control
0.00
0.00
0.00
*Mean of four replications; Values in parentheses are standard errors

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Table 5.5 - Larval mortality data of combination of emulsions against different

mosquito species.
Emulsion (%)
Karanj + castor + neem oil
0.001
0.0025
0.005
0.01
0.02
0.04
0.08
0.16
0.32
Untreated control
Karanj + castor oil
0.0025
0.005
0.01
0.02
0.04
0.08
0.16
0.32
0.64
Untreated control
Castor + neem oil
0.0025
0.005
0.01
0.02
0.04
0.08
0.16
0.32
0.64
Untreated control
Karanj + neem oil
0.0025
0.005
0.01
0.02
0.04
0.08
0.16
0.32
0.64
Untreated control

Corrected larval mortality at 24 hr (%)*

Culex quinque.
Aedes aegypti
Anoph. steph.
9(2.43)
18(2.93)
34(3.74)
44(3.83)
64(4.04)
76(4.32)
80(5.02)
96(5.08)
100
0.00

3(0.31)
15(0.48)
32(0.83)
49(1.03)
67(1.28)
78(1.93)
91(2.37)
98(3.28)
100
0.00

5(0.28)
19(0.3)
42(1.29)
58(2.03)
69(2.84)
78(2.93)
92(4.80)
100
0.00

1(0.04)
7(0.23)
24(0.48)
38(0.73)
49(0.83)
65(1.8)
79(2.93)
94(3.04)
100
0.00

3(0.32)
10(1.63)
19(1.90)
35(2.63)
55(3.84)
67(4.83)
81(5.25)
98(5.05)
100
0.00

7(0.05)
21(0.83)
33(0.73)
45(1.73)
59(1.83)
75(2.18)
87(3.67)
93(5.48)
100
0.00

6(1.28)
13(1.58)
25(2.04)
39(2.54)
55(3.19)
79(3.27)
92(4.03)
98(5.38)
100
0.00

9(0.08)
21(0.82)
31(1.20)
52(1.73)
69(2.19)
82(3.28)
92(4.10)
100
0.00

3(0.012)
7(0.27)
14(0.59)
25(0.92)
58(1.03)
75(1.28)
89(2.86)
100
0.00

4(0.03)
19(0.37)
35(0.92)
45(1.29)
67(1.73)
79(2.95)
85(3.74)
93(4.70)
100
0.00

9(1.03)
21(1.37)
42(2.04)
63(2.97)
74(3.28)
83(3.79)
92(4.02)
99(4.93)
100
0.00

7(0.76)
21(1.13)
33(1.85)
45(2.18)
59(3.14)
75(3.82)
87(4.58)
93(5.32)
100
0.00

*Mean of four replications; Values in parentheses are standard errors

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Chapter 5 Stable O/W Emulsions of Karanj, Castor and Neem Oil for Pesticide Applications.

Table 5.6 -Data on LC50, LC90, LC95 of karanj, neem and castor oil emulsions
against mosquito larvae.
LC50

LC90

LC95

Regression

(%)

(%)

(%)

equation

Culex quinquefasciatus

0.011

0.106

0.203

Y=1.319X + 7.561

0.979

Castor +

Aedes aegypti

0.011

0.064

0.107

Y=1.681X + 8.279

0.992

Neem

Anopheles stephensi

0.009

0.065

0.114

Y=1.581X + 8.078

0.978

Culex quinquefasciatus

0.024

0.193

0.350

Y=1.437X + 7.304

0.974

Aedes aegypti

0.015

0.098

0.167

Y=1.604X + 7.845

0.983

Anopheles stephensi

0.027

0.273

0.530

Y=1.289X + 7.005

0.940

Culex quinquefasciatus

0.040

0.237

0.394

Y=1.670X + 7.324

0.973

Aedes aegypti

0.033

0.188

0.312

Y=1.691X + 7.504

0.975

Anopheles stephensi

0.026

0.219

0.412

Y=1.348X + 7.169

0.993

Culex quinquefasciatus

0.024

0.141

0.234

Y=1.693X + 7.717

0.989

Aedes aegypti

0.018

0.134

0.236

Y=1.507X + 7.594

0.998

Anopheles stephensi

0.034

0.185

0.299

Y=1.768X + 7.575

0.986

Culex quinquefasciatus

0.106

0.948

1.785

Y=1.346X + 6.311

0.962

Aedes aegypti

0.094

0.769

1.409

Y=1.408X + 6.440

0.967

Anopheles stephensi

0.088

0.739

1.365

Y=1.390X + 6.462

0.997

Culex quinquefasciatus

0.063

0.268

0.400

Y=2.126X + 7.494

0.984

Aedes aegypti

0.109

0.506

0.788

Y=1.927X + 6.849

0.979

Anopheles stephensi

0.149

0.836

1.375

Y=1.712X + 6.413

0.987

Culex quinquefasciatus

0.215

1.527

2.690

Y=1.505X + 6.003

0.983

Aedes aegypti

0.182

0.857

1.993

Y=1.588X + 6.174

0.994

Anopheles stephensi

0.222

1.229

2.016

Y=1.724X + 6.125

0.977

Emulsion

Mosquito species

Karanj +

Karanj +
Neem

Karanj +
Castor

Neem +
Castor

Karanj

Neem

Castor

r2

LC50, LC90 and LC95are the concentration required to kill 50, 90 and 95% of the test
populations respectively,
LC50 / LC95 Fiducial limits with 95 % confidence,
Y = Probit mortality; X = Concentration

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Chapter 5 Stable O/W Emulsions of Karanj, Castor and Neem Oil for Pesticide Applications.

Table 5.7 - Increase in efficacy of karanj, neem and castor oil emulsion
combination over individuals against mosquito larvae.
Mosquito
Species

Culex quinq.
Aedes aegyp.
Anoph. steph

Culex quinq.
Aedes aegyp.
Anoph. steph

Culex quinq.
Aedes aegyp.
Anoph. steph

Culex quinq.
Aedes aegyp.
Anoph. steph

Fold increase of combination

Karanj + Neem + Castor oil emulsion (1:1:1)
Over karanj oil
Over Neem oil
Over castor oil emulsion
emulsion
emulsion
LC50

LC90

9.29
8.47
9.53

8.87
11.9
11.3

LC95

LC50

LC90

LC95

LC50

LC90

LC95

8.75 5.53 2.51 1.96 18.9 14.3

13.2
13.1 9.79 7.84 7.34 16.2 13.2
18.5
11.9 16.0 12.8 12.0 23.9 18.8
17.5
Karanj + Neem oil emulsion (1:1)
Over karanj oil emulsion
Over Neem oil emulsion
LC50
LC90
LC95
LC50
LC90
LC95
4.25
4.892
5.091
2.534
1.385
1.143
6.08
7.82
8.42
7.03
5.14
4.709
3.190
2.700
2.574
5.377
3.052
2.593
Karanj + castor oil emulsion (1:1)
Over karanj oil emulsion
Over castor oil emulsion
LC50
LC90
LC95
LC50
LC90
LC95
2.617
4.000
4.520
5.320
6.445
6.814
2.875
4.077
4.509
5.524
4.542
6.378
3.411
3.377
3.313
8.557
5.615
4.893
Neem + castor oil emulsion (1:1)
Over Neem oil emulsion
Over castor oil emulsion
LC50
LC90
LC95
LC50
LC90
LC95
2.544
1.896
11.48
8.689
10.78
11.48
5.804
3.772
3.336
9.654
6.385
8.437
4.283
4.516
4.587
6.375
6.644
6.724

5.3.8 House Fly Repellency

An UCR strain of Musca domestica is used in the repellency assay. A continuous
access to a mixture of skimmed milk powder / sugar (50:50) and water was given to flies.
A fly repellency chamber was made using a nylon-66 fibre net (having windows of ~2
mm diameter) and acrylic glue. The chamber dimensions were 20X20X10cm. The
multiple holes of the net maintained the air ventilation in the chamber. The chamber was
divided in four equal parts of dimension 10X10X10 cm. Four tightly woven cotton cloths
of 10X10cm were fixed at each divided floor part. Opposite corner cloths are used to
support test and placebo emulsion formulation. Test and placebo formulations (2mg/cm2)
were evenly distributed on the cotton cloth using a small pipette. The 50 flies (briefly
anaesthetized using CO2) were added to the chamber. Fly count was performed after the

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Chapter 5 Stable O/W Emulsions of Karanj, Castor and Neem Oil for Pesticide Applications.

interval of 1 hr and up to 3 days depending upon the fly positioning on the test and
placebo emulsion applied cotton cloth.
Fly count within the chamber showed that the flies were far away from the test
formulation applied cloth and were found to be very commonly distributed over the
untreated area of the chamber. 2 mg/cm2 concentration of oil emulsion showed a
promising fly repellency up to 72 hrs. The fly repellency potential of formulated oils were
found in the order of karanj > neem > castor. Table 5.8 shows the Musca domestica
repellency as a function of time.
Table 5.8 - Musca domestica repellency potential of formulated emulsions.
Musca domestica repellency % (after hrs)

Emulsion
(Formula)

12

24

48

72

Karanj (1)

100

100

100

100

100

100

92

Castor (9)

100

100

100

78

63

60

54

Neem(17)

100

100

100

100

100

85

70

5.3.9 Anti Maggot Activity (Exploratory study)

In a short term trial, the anti maggot potential of emulsion was tested in the case
of Myasis (Development of maggotted wound caused by the larva of Musca
domestica). The consistent spray of Karanj & Neem oil emulsion was found to be
effective remedy for expelling maggots from the wound. The process has some added
benefits like easy handing, minimal side effects and pain free alternative. Thus, the scope
of these formulations for detailed investigation is strongly recommended.
5.4 Conclusion
Karanj, castor and neem oil were successfully formulated as stable O/W emulsion.
The correlation between various physico-chemical parameters viz. oil content, surfactant
content, HLB number of surfactant blend, hardness and emulsion stability is established
to form a series of emulsion formulations. Rheological characterization has proved their
pseudoplastic flow nature thus making them a sprayable product. These emulsions have
shown a promising activity as mosquito larvicide, house fly repellent and maggoticide.

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