DESC:FORM 2

THE PATENTS ACT, 1970
(SECTION 39 OF 1970)
&
The Patents Rules, 2003

COMPLETE SPECIFICATION
(SECTION 10 and Rule 13)

AZADIRACHTIN FORMULATIONs AND PROCESS FOR PREPARING AND USE THEREOF

We, COROMANDEL INTERNATIONAL LIMITED,
an Indian company incorporated under Companies Act of 1956, having its principal place of business at Coromandel House, Sardar Patel Road, Secunderabad – 500 003, Telangana, India.

The following specification particularly describes the invention and the manner in which it is to be performed:
FIELD OF THE INVENTION
The present invention relates to stable Azadirachtin formulations comprising Azadirachtin as active ingredient and other agronomic additives.

The present invention relates to stable Azadirachtin Ready-To-Use formulation, comprising Azadirachtin as active ingredient, other agronomic additives selected from surfactant and one or more solvents.

The present invention also relates to stable Azadirachtin microemulsion formulation comprising Azadirachtin as active ingredient, other agronomic additives selected from a surfactant and one or more solvents.

The present invention more specifically relates to stable Azadirachtin microemulsion formulation comprising Azadirachtin as active ingredient, a surfactant and one or more solvents, wherein said formulation is devoid of additional plant extract/oils.

The present invention also relates to a process for the preparation of Azadirachtin formulations and other agronomic additives comprising steps of dissolving all ingredients and stirring to get clear solution.

BACKGROUND OF THE INVENTION
Azadirachtin is a chemical compound present in neem seeds. It is known to act
mainly as an anti-feedant and growth disruptor on a variety of insect species, while at the same time has minimal effects on beneficial arthropods. Consequently, Azadirachtin fulfills the criteria of an exceptionally good insecticide with widespread applicability.

Off late due to health and environmental concerns over the use of synthetic pesticides, there is a growing trend toward the use of natural pesticides which have little or no harmful environmental effects. Extensive research is carried out over the use of Azadirachtin in different forms such as neem powder, neem oil sprays, as well as formulated into emulsions.

US 5,110,591 A discloses water-in-oil microemulsion which, when diluted with water, forms a storage stable oil-in-water macroemulsion, said concentrated emulsion containing from about 50 to about 90 percent by weight of neem oil containing a pesticidally effective amount of azadirachtin and from about 50 to about 10 percent by weight of an emulsifier composition containing from about 40 to about 60 weight percent of nonionic emulsifying agent selected from the group consisting of sorbitan esters, ethoxylated and propoxylated mono- or diglycerides, lactated mono- or diglycerides, acetylated mono- or diglycerides, sugar-esters polyoxyethylene sorbitan esters containing from 4 to 20 polyoxyethylene groups, polyglycerol esters, or mixtures thereof, from about 20 to about 40 weight percent of polyhydric alcohol containing from 2 to 6 carbon atoms, and from about 10 to about 25 weight percent of water.

US 6,703,034 B2 discloses oil in water microemulsion comprising: at least one surfactant selected from the group consisting of non-ionic alkyl phenol ethoxylated surfactants; and a plurality of oil droplets comprising at least one oil including Neem Oil, wherein said microemulsion is primary short chain (C1 to C6) alcohol-free, ionic cosurfactant-free and stable upon dilution. Specific disclosed contain 20 grams of Neem Oil, 80.0 grams of a nonylphenol ethoxylate and sufficient water to total 200 grams which were blended until a nearly clear microemulsion concentrate was formed. The concentrate formed contained 10.0% Neem Oil and 40% emulsifier.

US 6,733,802 B1 discloses an emulsion comprising a biologically effective amount of azadirachtin in an essential oil, and a surface active agent, wherein said emulsion is made from naturally occurring materials and is devoid of synthetic compounds.

CN 103299992 A discloses Azadirachtin microemulsion, characterized by comprising the following components in mass ratio: Neem powder 0.5% ~ 5%; Solvents 18% ~ 25% ; Emulsifier 15% ~ 20% ; Synergist 1% ~ 5% and Deionized water 64.5 ~ 45%. The synergist is one or more of synergistic esters, synergistic sensitizers, synergistic amines, octachlorodipropyl ether, a formaldehyde aqueous solution with a mass concentration of 37%,

CN 1164176 C discloses azadirachtin microemulsion, extracted from the plant of the family Neem, it is characterized in that the azadirachtin in the seeds of the Azadirachta indica plant is used as the insecticidal active ingredient to provide the aid of the role of emulsifier, and the additives are added to make It is evenly dispersed in water to form a translucent azadirachtin microemulsion; the azadirachtin microemulsion is an oil-in-water type; the weight percentage composition is: azadirachtin 0.01% -85% emulsifier 0.05% -40% ; Cosolvent 0.01% -20% ; Stabilizer 0.01% -20% ; Synergist 0.05% -20% Penetration enhancer 0.1% -25% ; Antifreeze 0.1% -5% pH adjuster 0.01% -0.5% and Water 5% -80%; the emulsifiers are calcium alkylbenzene sulfonate, ammonium styrene polyoxyethylene ether sulfate, magnesium dialkyl ether disulfonate, alkyl ammonium salt, triethanolamine salt, benzyl biphenol polyoxyethylene Ether, phenethylphenol polyoxyethylene polyoxypropylene ether, nonylphenol polyoxyethylene ether, alkylphenol polyoxyethylene ether formaldehyde condensate, biphenol polyoxyethylene ether, the stabilizer is 3-chloro-1,2-epoxypropane, butyl glycidyl ether, phenyl glycidyl ether, tolyl glycidyl ether, polyvinyl glycol diglycidyl ether, sodium sorbate.

World Journal of Pharmaceutical Research 2019, Volume 8, Issue 3, 1507-1517 discloses neem oil Microemulsion system, which contains Neem oil, Lemon grass oil, Surfactants and water. The surfactants used are Atlox 4896 and nonylphenol 13. It is mentioned in this article that addition of 4% lemon grass oil, stability of Azadirachtin in neem oil microemulsion system improved significantly.

There are few Azadirachtin RTU products in the market which contain Neem oil (0.9%), potassium salts of fatty acid (1.0%); sodium benzoate and water (98.1%). However, all Neem oil based products are not stable and are not suitable for long-term applications.

In view of the importance of Azadirachtin as an acaricide or fungicide there is need in the art to provide environmentally safer and stable fungicidal formulations. The inventors of the present invention have developed new Azadirachtin microemulsion formulation with longer stability by using novel surfactant 2-ethyl hexanol ethylene oxide-propylene oxide for stabilizing the emulsion.

OBJECTIVE OF INVENTION
The main objective of the present invention is to provide a stable Azadirachtin formulations comprising Azadirachtin as active ingredient and other agronomic additives.

Another objective of the present invention is to provide a stable Azadirachtin Ready-To-Use formulation, comprising Azadirachtin as active ingredient, other agronomic additives selected from a surfactant and one or more solvents.

Another objective of the present invention is to provide a stable Azadirachtin microemulsion formulation, comprising Azadirachtin as active ingredient, other agronomic additives selected from a surfactant and one or more solvents.

Still another objective of the present invention is to provide a stable Azadirachtin Ready-To-Use formulation, comprising Azadirachtin as active ingredient, a surfactant and solvents, wherein said formulation is devoid of additional plant extract/oils.

Still another objective of the present invention is to provide stable Azadirachtin microemulsion formulation, comprising Azadirachtin as active ingredient, a surfactant and solvents, wherein the said formulation is devoid of additional plant extract/oils.

Yet another objective of the present invention is to provide to a process for the preparation of stable Azadirachtin formulations and other agronomic additives comprising steps of dissolving all ingredients and stirring to get clear solution.

SUMMARY OF INVENTION
Accordingly, the present invention provides stable Azadirachtin formulations comprising Azadirachtin as active ingredient and other agronomic additives.

In one embodiment, the present invention provides a stable Azadirachtin Ready-To-Use formulation, comprising Azadirachtin as active ingredient, other agronomic additives selected from a surfactant and one or more solvents.

In another embodiment, the present invention provides a stable Azadirachtin microemulsion formulation, comprising Azadirachtin as active ingredient, other agronomic additives selected from a surfactant and one or more solvents.

In another embodiment, the present invention provides a stable Azadirachtin Ready-To-Use formulation, comprising Azadirachtin as active ingredient, a surfactant, solvents, wherein said formulation is devoid of additional plant extract/oils.

In another embodiment, the present invention provides a stable Azadirachtin microemulsion formulation, comprising Azadirachtin as active ingredient, a surfactant, solvent, wherein said formulation is devoid of additional plant extract/oils.

In another embodiment, the present invention provides a stable Azadirachtin microemulsion formulation, comprising Azadirachtin as active ingredient, a surfactant, propylene carbonate as solvent, wherein said formulation is devoid of additional plant extract/oils.

In another embodiment, the present invention provides a stable Azadirachtin microemulsion formulation comprising:
(a) Azadirachtin as active ingredient,
(b) 2-ethyl hexanol ethylene oxide-propylene oxide as surfactant,
(c) propylene carbonate as solvent,
wherein said formulation is devoid of additional plant extract/oils.

In another embodiment, the present invention provides a stable Azadirachtin microemulsion formulation comprising:
(a) Azadirachtin as active ingredient,
(b) 2-ethyl hexanol ethylene oxide-propylene oxide as surfactant,
(c) propylene carbonate and demineralised water as solvent,
wherein said formulation is devoid of additional plant extract/oils.

In another embodiment, the present invention provides a stable Azadirachtin microemulsion formulation comprising:
(a) Azadirachtin in the range of 0.01% to 15% w/w, and
(b) ethoxylated sorbitan monolaurate in the range of 10% to 99% w/w.

In another embodiment, the present invention provides a stable Azadirachtin microemulsion formulation comprising:
(a) Azadirachtin in the range of 0.01% to 15% w/w,
(b) 2-ethyl hexanol ethylene oxide-propylene oxide in the range of 10% to 60% w/w, and
(c) propylene carbonate in the range of 40% to 90% w/w.

In another embodiment, the present invention provides a stable Azadirachtin 1.2% microemulsion formulation comprising:
(a) Azadirachtin extract in the range of 0.01% to 15% w/w,
(b) 2-ethyl hexanol ethylene oxide-propylene oxide in the range of 10% to 60% w/w, and
(c) propylene carbonate in the range of 40% to 90% w/w.

In another embodiment, the present invention provides a stable Azadirachtin microemulsion formulation comprising:
(a) Azadirachtin in the range of 0.01% to 15% w/w,
(b) 2-ethyl hexanol ethylene oxide-propylene oxide in the range of 10% to 60% w/w,
(c) propylene carbonate in the range of 40% to 90% w/w, and
(d) demineralised water in the range of 5% to 20% w/w.

In another embodiment, the present invention provides a process for the preparation of Azadirachtin formulations comprising steps of dissolving all ingredients and stirring to get clear solution.

In another embodiment, the present invention provides a process for the preparation of Azadirachtin microemulsion, wherein said process comprising steps of:
(a) dissolving Azadirachtin in surfactant, and
(b) stirring the mixture for 30 min to obtain clear formulation without any undissolved matter or haziness.
In another embodiment, the present invention provides a process for the preparation of Azadirachtin microemulsion, wherein said process comprising steps of:
(a) dissolving Azadirachtin technical in solvent,
(b) adding surfactant, and
(c) stirring the mixture for 30 min to obtain clear formulation without any undissolved matter or haziness.

In another embodiment, the present invention provides a process for the preparation of Azadirachtin microemulsion, wherein said process comprising steps of:
(a) dissolving Azadirachtin in ethoxylated sorbitan monolaurate, and
(b) stirring the mixture for 30 min to obtain clear formulation without any undissolved matter or haziness.

In another embodiment, the present invention provides a process for the preparation of Azadirachtin microemulsion, wherein said process comprising steps of:
(a) dissolving Azadirachtin technical in propylene carbonate,
(b) adding 2-ethyl hexanol ethylene oxide-propylene oxide, and
(c) stirring the mixture for 30 min to obtain clear formulation without any undissolved matter or haziness.

In another embodiment, the present invention provides a process for the preparation of Azadirachtin microemulsion, wherein said process comprising steps of:
(a) dissolving Azadirachtin technical in propylene carbonate and de-mineralized water,
(b) adding 2-ethyl hexanol ethylene oxide-propylene oxide, and
(c) stirring the mixture for 30 min to obtain clear formulation without any undissolved matter or haziness.

In still another embodiment, the present invention provides a process for the preparation of Azadirachtin RTU formulation, wherein said process comprising steps of:
(a) dissolving Azadirachtin technical in propylene carbonate,
(b) adding ethylene oxide-propylene oxide sorbitan monolaurate and
(c) the mixture was stirred for 30 minutes to obtain clear composition without any undissolved matter or haziness.

DETAILED DESCRIPTION OF THE INVENTION
The term "comprising", which is synonymous with "including", "containing", or "characterized by" here is defined as being inclusive or open-ended, and does not exclude additional, unrecited elements or method steps, unless the context clearly requires otherwise.

The present invention specifically provides stable Azadirachtin Ready-To-Use formulation comprising:
a) Azadirachtin technical in the range of 0.01% to 15 %,
b) surfactant in the range of 10% to 99.8%,
c) one or more solvents.

The present invention specifically provides stable Azadirachtin microemulsion formulation comprising:
a) Azadirachtin technical in the range of 1% to 15 %,
b) surfactant in the range of 10% to 99%,
c) one or more solvents.

The present invention specifically provides stable Azadirachtin 1.2% microemulsion formulation comprising :
a) Azadirachtin extract in the range of 0.01% to 15% w/w,
b) 2-ethyl hexanol ethylene oxide-propylene oxide in the range of 10% to 60% w/w, and
c) propylene carbonate in the range 40% to 90% w/w.

The stable Azadirachtin formulations of the present invention are devoid of any additional plant extract/oils.

The term "agrochemically effective amount" is that quantity of active agent, applied in any amount which will provide the required control of diseases. The particular amount is dependent upon many factors including, for example, type of formulations, the crop, disease sought to be controlled and environmental conditions. The selection of the proper quantity of active agent to be applied, however, is within the expertise of one skilled in the art.

In an embodiment of the present invention, the formulation comprises of: a Azadirachtin technical present in a range from 0.01% to 15% (w/w); surfactant present in a range from 10% to 99.8% (w/w). The Azadirachtin technical suitable for use in the present invention has minimum percentage purity in the range of 30 to 40%. The Azadirachtin technical suitable for use in the present invention has base value in the range of 0.01 to 2.0%.

In another embodiment of the present invention, the formulation comprises from 10% to 99.8% (w/w) of surfactant which also acts as emulsifier and dispersant. Surfactants are used not only to improve dispersion and to emulsify the product, but also to increase suspension stability, wettability, penetration and translocation, and to provide the mixing ability and suspension/emulsion stability of a product after dilution.

As surfactants, anionic, cationic, non-ionic and ampholytic surfactants and mixtures thereof can be used. Suitable surfactants are, for example, nonionic emulsifiers and dispersants, such as: polyalkoxylated, preferably polyethoxylated, saturated and unsaturated aliphatic alcohols, having 8 to 24 carbon atoms in the alkyl radical, which is derived from the corresponding fatty acids or from petrochemical products, and having 1 to 100, preferably 4 to 40, ethylene oxide units (EO); polyalkoxylated, preferably polyethoxylated; polyalkoxylated, preferably polyethoxylated, alkylphenols having one or more alkyl radicals, such as, for example, nonylphenol or tri-sec- butylphenol, and a degree of ethoxylation of between 2 and 40, preferably from 4 to 20; polyalkoxylated, preferably polyethoxylated, hydroxyl-fatty acids or glycerides which contain hydroxyl-fatty acids, such as, for example, castor oil, having a degree of ethoxyiation of between 10 and 80; sorbitan or sorbitol esters with fatty acids or polyalkoxylated, preferably polyethoxylated, sorbitan or sorbitol esters; polyalkoxylated, preferably polyethoxylated, amines; di- and tri-block copolymers, for example from alkylene oxides, for example from ethylene oxide and propylene oxide, having average molar masses between 200 and 8000 g/mol, preferably from 1000 to 4000; alkylpolyglycosides or polyalkoxylated, preferably polyethoxylated, alkylpolyglycosides. Preferred nonionic surfactants are polyethoxylated alcohols, preferably from renewable resources, such as ethoxylated (4-8 EO) C12-C14 natural alcohol; polyethoxylated triglycerides which contain hydroxyfattyacids. Preferably non-ionic surfactant is Lutensol (Ethoxylate of alkyl polyethylene glycol ether). Lutensol® is a non-ionic surfactant. It is alkyl polyethylene glycol ether made from a C10-Guerbet alcohol and ethylene oxide which contains also higher alkylene oxide in small amounts.

Anionic surfactants are for example: polyalkoxylated, preferably polyethoxylated, surfactants which are ionically modified, for example by conversion of the terminal free hydroxyl function of the alkylene oxide block into a sulfate or phosphate ester; polyelectrolytes, such as lignosulfonates, condensates of naphthalenesulfonate and formaldehyde, polystyrenesulfonate or sulfonated unsaturated or aromatic polymers; anionic esters of alkylpolyglycosides, such as alkylpolygiucosidesulfosuccinate or citrate; sulfosuccinates which are esterified once or twice with linear, or branched aliphatic, cycloaliphatic and/or aromatic alcohols, or sulfosuccinates which are esterifted once or twice with (poly)alkylene oxide adducts of alcohols. Preferred anionic surfactants are, for example, salts of aikylarylsulfonic and sulfosuccinic acids, and polyelectrolytes Examples of cationic and ampholytic surfactants are quanternary ammonium salts, alkyl amino acids, and betaine or imidazolineamphotensides and Ethoxylated Castor Oil, Tristyrylphenol Alkoxylate.

Preferred surfactants used in the composition of the present invention are sorbitane monolaurate, ethoxylated sorbitan monolaurate (Emulsogen 4156), polyoxyethylene (20) sorbitan monolaurate (Polysorbate 20), Polyoxyethylene (20) sorbitan monooleate (Polysorbate 80) and 2-ethyl hexanol ethylene oxide-propylene oxide (ECOSURF EH-6).

The solvent used in the compositions of the present invention include Propylene carbonate, water or mixtures thereof.

The compositions of the present invention do not require essential oil. The surfactant used in the present invention itself acts as good stabilizer, readily solubilizes Azadirachtin, act as an inbuilt adjuvant, thereby minimizes the Azadirachtin degradation. Due to the stability provided by Surfactant, the compositions of the present invention show stability on storage at temperatures of 54°C for up to 2 weeks and also stable at ambient temperature for up to 2 years.

Azadirachtin 1.2% ME formulation is stable for 2 years whereas Azadirachtin with other formulations such as EC are not stable.

The fungicidal compositions according to the present invention possess advantageous curative, preventive and antisporulant fungicidal activity to protect plants, fruit and seeds. The present composition can be used to protect plants or parts of plants, e.g. fruit, blossoms, flowers, foliage, stems, roots, cuttings, tubers of plants or culture plants infected, harmed or destroyed by micro-organisms, whereby later-growing parts of plants are protected against such micro-organisms. Application may be made by ground or aerial spray equipment.

A particular mode of administering an antifungal composition of the present invention is the administration to the aboveground parts of plants, in particular to the leaves thereof (leaf-application). The number of applications and the administered doses are chosen in accordance with the biological and climatic conditions of life of the causative agent. The antifungal compositions though, can also be applied to the soil and get into the plants through the root system (systemic activity), in case the locus of the plants is sprayed with a liquid composition or if the components are added to the soil in a solid formulation e.g. in the form of granulate (soil application).

Although the invention has been described above with reference to the disclosed embodiments, those skilled in the art will readily appreciate that the specific embodiments detailed are only illustrative of the invention. It should be understood that various modifications can be made without departing from the spirit of the invention. Accordingly, the invention is limited only by the following claims.

The following examples describes the nature of the invention which are given only for the purpose of illustrating the present invention in more detail and are not limitative and relate to solutions, which have been particularly effective on bench scale.

EXAMPLES
Example 1: Azadirachtin RTU composition
S.No Ingredients/Raw Materials Role Quantity (w/w %)
1. Azadirachtin Technical @35.2%purity, Base@0.02% Technical 0.057
2. Propylene carbonate Solvent 49.97
3. ethylene oxide-propylene oxide sorbitane monolaurate Surfactant 49.97
Total 100

Manufacturing process:
Azadirachtin technical was dissolved in propylene carbonate, stirred for 30 minutes. Ethylene oxide-propylene oxide sorbitane monolaurate was added to the above mixture and stirred for 30 minutes to get clear solution without any undissolved matter or haziness.

Example 2: Azadirachtin RTU composition
S.No Ingredients/Raw Materials Role Quantity (w/w %)
1. Azadirachtin Technical @35.2%purity, Base@0.02% Technical 0.057
2. Propylene carbonate Solvent 64.94
3. 2-ethyl hexanol ethylene oxide-propylene oxide (ECOSURF EH-6) Surfactant 25.0
4. De-mineralized water Solvent 10.0
Total 100

Manufacturing process:
Azadirachtin technical was dissolved in Propylene carbonate, stirred for 30 minutes. De-mineralized water was added to the above mixture. 2-ethyl hexanol ethylene oxide-propylene oxide (ECOSURF EH-6) was added to the above mixture and stirred for 30 minutes to get clear solution without any undissolved matter or haziness.

Example 3: Azadirachtin Microemulsion composition
S.No Ingredients/Raw Materials Role Quantity (w/w %)
1. Azadirachtin (a) 35.21% (b) 1.2% Technical 3.41
2. Propylene carbonate (oil) Solvent 61.6
3. 2-ethyl hexanol ethylene oxide-propylene oxide (ECOSURF EH-6) Emulsifier / Surfactant 25.0
4. Demineralised water Solvent 10.0
Total 100

Manufacturing process:
Azadirachtin technical was dissolved in Propylene carbonate, stirred for 30 minutes. De-mineralized water was added to the above mixture. 2-ethyl hexanol ethylene oxide-propylene oxide (ECOSURF EH-6) was added to the above mixture and stirred for 30 minutes to get clear solution without any undissolved matter or haziness.

Stability data of Example 3 composition
Physicochemical test RT sample@ 7day 7 AHS sample@54°C
Appearance Clear transparent liquid, without any haziness Clear transparent liquid, without any haziness. Slight dark in colour as compared to RT
A.I content(%w/w) 1.24% 0.99%
Emulsion Stability after 24hr Clear column without any settling matter Clear column without any settling matter

Example 4: Azadirachtin Microemulsion composition:
S.No Ingredients/Raw Materials Role Quantity (w/w %)
1. Azadirachtin (a) 35.21% (b) 1.2% Technical 3.41
2. Propylene carbonate (oil) Solvent 71.6
3. 2-ethyl hexanol ethylene oxide-propylene oxide (ECOSURF EH-6) Emulsifier / Surfactant 25.0
Total 100

Manufacturing process:
Azadirachtin technical was dissolved in Propylene carbonate, stirred for 30 minutes. 2-ethyl hexanol EO-PO (ECOSURF EH-6) was added to the above mixture and stirred for 30 minutes to get clear solution without any undissolved matter or haziness.

Stability data of Example 4 composition
Physicochemical test RT sample@ 7day 7 AHS sample@54°C
Appearance Clear transparent liquid, without any haziness Clear transparent liquid, without any haziness. Slight dark in colour as compared to RT
A.I content(%w/w) 1.14% 0.94%
Emulsion Stability after 24hr Clear column without any settling matter Clear column without any settling matter

Example 5: Azadirachtin Microemulsion composition
S.No Ingredients/Raw Materials Role Quantity (w/w %)
1. Azadirachtin (a) 35.21% (b) 1.2% Technical 3.70
2. Propylene carbonate (oil) Solvent 61.3
3. 2-ethyl hexanol ethylene oxide-propylene oxide (ECOSURF EH-6) Emulsifier / Surfactant 25.0
4. Demineralised water Solvent 10.0
Total 100

Manufacturing process:
Azadirachtin technical was dissolved in Propylene carbonate, stirred for 30 minutes. De-mineralized water was added to the above mixture. 2-ethyl hexanol ethylene oxide-propylene oxide (ECOSURF EH-6) was added to the above mixture and stirred for 30 minutes to get clear solution without any undissolved matter or haziness.

Stability data of Example 5 composition
Physicochemical test Specification RT 14days AHS 14day@54°C
Appearance Clear
solution Clear solution Clear
solution
A.I content(%w/w) 1.02-1.38% 1.28% 1.08%
Emulsion Stability after 24hr Clear solution without any settling Clear solution without any settling matter Clear solution without any settling matter

Example 6: Azadirachtin Microemulsion composition:
S.No Ingredients/Raw Materials Role Quantity (w/w %)
1. Azadirachtin (a) 35.21% (b) 1.3% Technical 3.70
2. Propylene carbonate Solvent 71.3
3. 2-ethyl hexanol ethylene oxide-propylene oxide (ECOSURF EH-6) Emulsifier 25.0
Total 100

Manufacturing process:
Azadirachtin technical was dissolved in Propylene carbonate, stirred for 30 minutes. 2-ethyl hexanol ethylene oxide-propylene oxide (ECOSURF EH-6) was added to the above mixture and stirred for 30 minutes to get clear solution without any undissolved matter or haziness.

Stability data of Example 6 composition
Physicochemical test Specification RT 14days AHS 14day@54°C
Appearance Clear solution Clear solution Clear solution
A.I content(%w/w) 1.02-1.38% 1.27% 1.06%
Emulsion Stability after 24hr Clear solution without any settling Clear solution without any settling matter Clear solution without any settling matter

Example 7: Azadirachtin Microemulsion composition:
S.No Ingredients/Raw Materials Role Quantity (w/w %)
1. Azadirachtin (a) 35.21% (b) 1.2% Technical 3.40
2. ethoxylated sorbitan monolaurate (Emulsogen 4156) Emulsifier 96.59
Total 100

Manufacturing process:
Azadirachtin technical was dissolved in ethoxylated sorbitan monolaurate Emulsogen 4156 and stirred for 30 minutes to get clear solution without any undissolved matter or haziness.

Example 8: Azadirachtin 1.2 Microemulsion composition:
S.No Ingredients/Raw Materials Role Quantity (w/w %)
1. Azadirachtin (a) 35.21% (b) 1.2% Technical 3.40
2. Polyoxyethylene (20) sorbitan monolaurate (Polysorbate 20) Emulsifier 96.59
Total 100

Manufacturing process:
Azadirachtin technical was dissolved in polyoxyethylene sorbitan monolaurate Polysorbate 20 and stirred for 30 minutes to get clear solution without any undissolved matter or haziness.

Example 9: Azadirachtin 1.2 Microemulsion composition:
S.No Ingredients/Raw Materials Role Quantity (w/w %)
1. Azadirachtin (a) 35.21% (b) 1.2% Technical 3.40
2. Polyoxyethylene (20) sorbitan monooleate (Polysorbate 80) Emulsifier 96.59
Total 100

Manufacturing process:
Azadirachtin technical was dissolved in polyoxyethylene sorbitan monooleate (Polysorbate 80) and stirred for 30 minutes to get clear solution without any undissolved matter or haziness.

Phytotoxicity of Azadirachtin Microemulsion formulations on Mulberry

The plants with new shoots were selected and tagged with treatment details. The test solutions were prepared as per the protocol and sprayed using hand sprayer. The plants were drenched thoroughly. Untreated control plants were given with water spray. Observations were recorded 24 hours after treatment and up to seven days (1st day, 3rd day, 5th day and 7th day). The parameters such as scorching, wilting, necrosis, vein clearing, epinasty and hyponasty.

Table 1: Phytotoxicity of AZADIRACHTIN Microemulsion formulations on Mulberry
S.No Treatments with dosage Phytotoxicity symptoms Period of observation
1 DAT
3 DAT 5 DAT 7 DAT
1 Azadirachtin Microemulsion - (3.9 ml/lit) Injury on leaf tips 0 0 0 0
Injury on leaf surface 0 0 0 0
Leaf wilting 0 0 0 0
Necrosis 0 0 0 0
Epinasty 0 0 0 0
Hyponasty 0 0 0 0
2 Azadirachtin Microemulsion - (7.8 ml/lit) Injury on leaf tips 0 0 0 0
Injury on leaf surface 0 0 0 0
Leaf wilting 0 0 0 0
Necrosis 0 0 0 0
Epinasty 0 0 0 0
Hyponasty 0 0 0 0
3 Azadirachtin Microemulsion - (12 ml/lit) Injury on leaf tips 0 0 0 0
Injury on leaf surface 0 0 0 0
Leaf wilting 0 0 0 0
Necrosis 0 0 0 0
Epinasty 0 0 0 0
Hyponasty 0 0 0 0
4 Un treated control Injury on leaf tips 0 0 0 0
Injury on leaf surface 0 0 0 0
Leaf wilting 0 0 0 0
Necrosis 0 0 0 0
Epinasty 0 0 0 0
Hyponasty 0 0 0 0
*DAT- Days after Treatment

Based on the observation, it can be concluded that the Phytotoxicity tests of Azadirachtin Microemulsion at doses such as 3.9 ml/lit, 7.8 ml/lit and 12 ml/lit, performed in Mulberry plants (Morus sp.) does not shows phytotoxic symptoms like Scorching, Wilting, Necrosis, Vein clearing, Epinasty and Hyponasty. Hence it can be concluded that Azadirachtin Microemulsion formulation is safe to plants.

Bio efficacy evaluation of Azadirachtin microemulsion (ME) formulation:

Table 2: Treatment details
Treatment details
Treatment Dose (ml/lit)
T1 – AZADIRACHTIN ME 3.9
T2 - AZADIRACHTIN ME 7.8
T3 - AZADIRACHTIN ME 12
T4 – AzaMax 3.9
T5 – AzaMax 7.8
T6 – AzaMax 12
T7 - Botanical Check – Neemazal T/S 3
T8 - Un-Treated control (water spray) -

The treatment product applications were given during experimentation. Spray was given with Hand operated sprayer using spray volume of 500 liters per hectare. After 30 mins of spray once after the plant become dry. Whitefly will be released in plant and locked using clip cage (5 Adults/Cage/leaf) as top, middle, and bottom. Observation on live Adults will be monitored on 1 DAT, 3 DAT, 5 DAT and 7 DAT (Day After Treatment). The per cent control against Whitefly over untreated control was also worked out.
Table. 3: Trial 1- Evaluation of Azadirachtin ME formulation against Whitefly (Bemisia tabaci) on cotton
No of live adults/Leaf (or) Clip cage
Treatment details Dose (ml/l) (0 Day) 1 DAT 3 DAT 5 DAT 7 DAT Mean % ROC


T1 – AZADIRACHTIN ME 3.9 5 3.89 3.22 1.56 0.33 2.25 (1.59) 55.00
T2 - AZADIRACHTIN ME 7.8 5 3.67 2.67 1.22 0.22 1.94 (1.49) 61.11
T3 - AZADIRACHTIN ME 12 5 3.22 2.11 0.78 0.00 1.53 (1.34) 69.44
T4 – AzaMax 3.9 5 4.22 3.33 1.78 0.44 2.44 (1.65) 51.11
T5 – AzaMax 7.8 5 3.67 2.67 1.33 0.33 2.00 (1.52) 60.00
T6 – AzaMax 12 5 3.22 2.56 1.00 0.00 1.69 (1.40) 66.11
T7 - Botanical Check – Neemazal T/S 3 5 3.89 2.78 1.56 0.44 2.17 (1.57) 56.67
T8 - Un-Treated control (water spray) - 5 5.00 5.00 5.00 5.00 5.00 (2.34) -
Three leaves (Top, Middle, Bottom /Plant), Figures in Parentheses are square root transformed.
*DAT-Days After Treatment

The Semi field experiment results revealed that the mean reduction of Whitefly adult population was higher in AZADIRACHTIN ME when compared with AzaMax and Neemazal T/S.
Table.4: Trial 2- Evaluation of Azadirachtin ME formulation against Whitefly (Bemisia tabaci) on cotton
No of live adults/Leaf (or) Clip cage
Treatment details Dose (ml/l) (0 Day) 1 DAT 3 DAT 5 DAT 7 DAT Mean % ROC


T1 – AZADIRACHTIN ME 3.9 5 3.44 2.67 1.33 0.44 1.97 (1.52) 60.56
T2 - AZADIRACHTIN ME 7.8 5 3.22 2.22 0.89 0.33 1.67 (1.41) 66.67
T3 - AZADIRACHTIN ME 12 5 3.11 1.78 0.33 0.22 1.36 (1.29) 72.78
T4 – AzaMax 3.9 5 3.67 2.78 1.56 0.56 2.14 (1.57) 57.22
T5 – AzaMax 7.8 5 3.22 2.22 1.33 0.44 1.81 (1.47) 63.89
T6 – AzaMax 12 5 2.67 1.78 0.67 0.11 1.31 (1.28) 73.89
T7 - Botanical Check – Neemazal T/S 3 5 3.56 2.56 0.89 0.33 1.83 (1.46) 63.33
T8 - Un-Treated control (water spray) - 5 5.00 5.00 5.00 5.00 5.00 (2.34) -
Three leaves (Top, Middle, Bottom /Plant), Figures in Parentheses are square root transformed. *DAT-Days After Treatment

The Semi field experiment results revealed that the mean reduction of Whitefly adult population was higher in AZADIRACHTIN ME when compared with AzaMax and Neemazal T/S.
Table.5: Evaluation of Azadirachtin ME formulation against Whitefly (Bemisia tabaci) on cotton plant
No of live adults/Leaf (or) Clip cage
Treatment details Dose (ml/lit) 0 Day Trial 1 Trial 2 Mean % ROC


T1 – AZADIRACHTIN ME 3.9 5 2.25 (1.59) 1.97 (1.52) 2.11 57.8
T2 - AZADIRACHTIN ME 7.8 5 1.94 (1.49) 1.67 (1.41) 1.81 63.9
T3 - AZADIRACHTIN ME 12 5 1.53 (1.34) 1.36 (1.29) 1.45 71.1
T4 – AzaMax 3.9 5 2.44 (1.65) 2.14 (1.57) 2.29 54.2
T5 – AzaMax 7.8 5 2.00 (1.52) 1.81 (1.47) 1.91 61.9
T6 – AzaMax 12 5 1.69 (1.40) 1.31 (1.28) 1.50 70.0
T7 - Botanical Check – Neemazal T/S 3 5 2.17 (1.57) 1.83 (1.46) 2.00 60.0
T8 - Un-Treated control (water spray) - 5 5.00 (2.34) 5.00 (2.34) 5.00
Three leaves (Top, Middle, Bottom /Plant), Figures in Parentheses are square root transformed.
*DAT-Days After Treatment

The Semi field experiment results revealed that the mean reduction of Whitefly adult population was higher in AZADIRACHTIN ME when compared with AzaMax and Neemazal T/S.

,CLAIMS:WE CLAIM:

1. A stable Azadirachtin 1.2% microemulsion formulation comprising Azadirachtin as active ingredient, a surfactant, wherein the said formulation is devoid of additional plant extract/oils.
2. The formulation as claimed in claim 1, wherein said surfactants are selected from sorbitane monolaurate ethylene oxide-propylene oxide, ethoxylated sorbitan monolaurate and 2-ethyl hexanol ethylene oxide-propylene oxide.
3. The formulation as claimed in claim 1, wherein said formulation contains solvent selected from propylene carbonate, water or mixtures thereof.
4. The formulation as claimed in claims 1–2, wherein said Azadirachtin microemulsion formulation comprising:
(a) Azadirachtin in the range of 0.01% to 15% w/w, and
(b) ethoxylated sorbitan monolaurate in the range of 10% to 99% w/w.
5. An Azadirachtin 1.2% microemulsion formulation comprising:
a) Azadirachtin extract in the range of 0.01% to 15% w/w,
b) 2-ethyl hexanol ethylene oxide-propylene oxide in the range of 10% to 60% w/w, and
c) propylene carbonate in the range of 40% to 90% w/w.
6. The formulation as claimed in claims 1 - 5, wherein said Azadirachtin microemulsion formulation comprising:
(a) Azadirachtin in the range of 0.01% to 15% w/w,
(b) 2-ethyl hexanol ethylene oxide-propylene oxide in the range of 10% to 60% w/w,
(c) propylene carbonate in the range of 40% to 90% w/w, and
(d) demineralised water in the range of 5% to 20% w/w.
7. The process for the preparation of formulation as claimed in claims 1 – 6, wherein said process comprising steps of:
(c) dissolving Azadirachtin in surfactant, and
(d) stirring the mixture for 30 min to obtain clear formulation without any undissolved matter or haziness.
8. The process for the preparation of formulation as claimed in claim 7, wherein said process comprising steps of:
(a) dissolving Azadirachtin technical in solvent,
(b) adding surfactant, and
(c) stirring the mixture for 30 min to obtain clear formulation without any undissolved matter or haziness.
9. The process for the preparation of formulation as claimed in claim 5, wherein said process comprising steps of:
(a) dissolving Azadirachtin technical in propylene carbonate,
(b) adding 2-ethyl hexanol ethylene oxide-propylene oxide, and
(c) stirring the mixture for 30 min to obtain clear formulation without any undissolved matter or haziness.
10. The process for the preparation of formulation as claimed in claims 6, wherein said process comprising steps of:
(a) dissolving Azadirachtin technical in propylene carbonate and de-mineralized water,
(b) adding 2-ethyl hexanol ethylene oxide-propylene oxide, and
(c) stirring the mixture for 30 min to obtain clear formulation without any undissolved matter or haziness.

Dated this Sixth (6th) day of January, 2022

__________________________________
Dr. S. Padmaja
Agent for the Applicant
IN/PA/883