Description:SYNERGISTIC INSECTICIDAL COMPOSITION
FIELD OF INVENTION
The present invention relates to a novel, synergistic insecticidal composition for effective control of insect pests in agricultural crops, particularly in cotton. The present invention further relates to a process for preparing said insecticidal composition.

BACKGROUND OF THE INVENTION
Insecticides are chemicals that are used to control insects by killing them or preventing them from engaging in undesirable or destructive behaviors. They are of varied types depending upon structure and mode of action. Lately, the application of combination of insecticides has emerged as an attractive agricultural practice to ensure that the chemical load in the environment is reduced. A combination of agricultural pesticides may be applied sequentially, as a tank mix application or as a ready-to-use composition.

A tank mix of pesticides in many cases results in a heterogenous solution and suffers from several other problems like difficult handling, sedimentation, etc. Hence a pre-mix composition is much preferred. However, it is a known fact that not all actives are compatible with each other, and therefore experimentation is required to develop a stable, synergistic, efficacious pesticidal composition or formulation.

Cotton (Gossypium spp.) is a key cash crop worldwide. It is prone to damage from various pests, including Jassids (Amrasca biguttula biguttula) and Whitefly (Bemisia tabaci), which cause substantial economic losses. These pests inflict harm by sucking plant sap, leading to stunted growth, chlorosis, and reduced yields. Moreover, Whitefly acts as a vector for viral diseases, further aggravating crop damage (Nagrare et al., 2019). The increasing resistance of these pests to conventional insecticides necessitates the development of innovative pest control measures.

To address this problem, the present inventors have developed the novel insecticidal active combination of Flonicamid and Spirotetramat along with polysaccharides or modified polysaccharides mixture that shows potential in not only addressing the pest control but also stimulates plant growth and enhances yield. Flonicamid is a selective feeding disruptor that effectively targets sap-sucking pests (Morita et al., 2007), while Spirotetramat, a systemic ketoenol insecticide, disrupts lipid biosynthesis and provides prolonged pest control (Bruck et al., 2009). The addition of polysaccharides or modified polysaccharides enhances the formulation's adhesion, efficacy, and resistance to environmental factors, ensuring consistent performance in field conditions. Further, polysaccharides are known to have bio stimulant effect on crops and plants.

The present invention further aims to provide an insecticidal composition that is synergistic in action. More particularly, the present invention aims at providing a composition wherein the insecticidal efficacy of the composition is significantly enhanced as compared to the sum of the activities of the solo active compounds when applied alone.

OBJECT OF THE INVENTION
It is an object of the present invention to provide a novel, stable, synergistic, insecticidal composition comprising a) Flonicamid b) Spirotetramat and c) polysaccharides or modified polysaccharides along with at least one agriculturally acceptable excipient.

It is yet another object of the present invention to provide a ready-to-use insecticidal composition that shows an enhanced bioefficacy at a reduced dosage as compared to insecticidal actives when applied individually.

It is yet another object of the present invention to provide an insecticidal composition that is highly effective in controlling sucking pests including but not limited to white fly, jassids, aphids, thrips in agricultural crops particularly in cotton.

It is yet another object of the present invention to provide an insecticidal composition that is environmentally safe and non-phytotoxic.

SUMMARY OF THE INVENTION
Accordingly, in one aspect, the present invention provides a novel, stable, synergistic, cost-effective insecticidal composition comprising a) Flonicamid b) Spirotetramat and c) polysaccharides or modified polysaccharides along with at least one agriculturally acceptable excipient.
In another aspect, the present invention provides a novel, stable, synergistic insecticidal composition that is highly effective in controlling sucking pests comprising whitefly, jassids, aphids and thrips in agricultural crops, particularly in cotton.

In yet another aspect, the present invention provides a process for preparing said insecticidal composition in the form of a suspension concentrate.

DETAILED DESCRIPTION OF THE INVENTION:
Those skilled in the art will be aware that the invention described herein is subject to variations and modifications other than those specifically described. It is to be understood that the invention described herein includes all such variations and modifications. The invention also includes all such steps, features, compositions and compounds referred to or indicated in this specification, individually or collectively, and all combinations of any two or more of said steps or features.

Unless otherwise specified, all terms used in disclosing the invention, including technical and scientific terms, have the meaning that is commonly understood by one of the ordinary skill in the art to which the invention belongs. For further guidance, term definitions may be included to better appreciate the teaching of the present invention.

As used herein, the term “plant” refers to any plant or part thereof including serial and subterranean parts of the plant. It is contemplated that the parts of the plant may be for example, flowers, fruits or vegetables, shoots, leaves, needles, stalks, stems, fruiting bodies, seeds also roots and that parts of the plants may or may not be attached to the remainder of the plant.

The term “locus” of a plant as used herein is intended to embrace the place on which the plants are growing, where the plant propagation materials of the plants are sown or where the plant propagation materials of the plants will be placed into the soil.

The term “plant propagation material” is understood to denote generative parts of a plant, such as seeds, vegetative material such as cuttings or tubers, roots, fruits, tubers, bulbs, rhizomes and parts of plants, germinated plants and young plants which are to be transplanted after germination or after emergence from the soil. These young plants may be protected before transplantation by a total or partial treatment by immersion.

It is to be understood that the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise.

It should be emphasized that the term “comprises/comprising” when used in this specification is taken to specify the presence of stated features, steps or components but does not preclude the presence or addition of one or more other features, steps, components or groups thereof.

The term "insects" as used herein, includes all organisms in the class "Insecta."

The term "Insecticidal" as used herein, refers to the ability of a pesticide to increase mortality or inhibit growth rate of insects.

The term "insecticidally effective amount" refers to an amount of an active that kills or inhibits the insect pest on which control is desired, in an amount not significantly toxic to the plant being treated.

The term “Polysaccharide” and “Modified Polysaccharides” can be used interchangeably.

The abbreviation "D" as used herein, has been used in the context of particle size distribution. D90 means that 90% of the total particles are smaller than this size.

The term “square” as used herein, in the context of cotton plants, refers to the flower bud, a precursor to the actual cotton flower and eventually the boll (the fruit containing the fibers).

The terms “weight percent”, “wt-%”, “percent by weight”, “% by weight” and variations thereof, as used herein, refer to the concentration of a substance as the weight of that substance divided by the total weight of the composition and multiplied by 100. It is understood that, as used here, “percent “, “%” and the like are intended to be synonymous with “weight percent”, “wt. %”, etc.
Abbreviations:
DM Demineralised water
RTU Ready to use
QS Quantum satis
A.I/ a.i Active ingredient
Ai/ha Active ingredient per hectare
DAA Days after application
RPM Revolutions per minute
SC Suspension concentrates

Features that are described and/or illustrated with respect to one embodiment may be used in the same way or in a similar way in one or more other embodiments and/or in combination with or instead of the features of the other embodiments.

Flonicamid (IUPAC name: N-cyanomethyl-4-(trifluoromethyl)nicotinamide) is a pyridine organic insecticidal compound discovered by Ishihara Sangyo Kaisha that inhibits feeding. It has a systemic and translaminar activity. Besides showing excellent aphicidal activity it gives a long-term control over sucking insect pests such as thrips, whiteflies, planthoppers, leafhoppers, plant bugs and mealybugs in fruit trees, cereals, rice, potatoes, cotton, vegetables, ornamentals and horticultural plants. Flonicamid has no negative impact on pollinating insects or natural enemies. Flonicamid also has a good toxicological, environmental and ecotoxicological profile.

Spirotetramat, IUPAC name [3-(2,5-dimethylphenyl)-8-methoxy-2-oxo-1-azaspiro[4.5]dec-3-en-4-yl] ethyl carbonate is a keto-enol insecticide which is the only modern 2-way systemic insecticide in the world i.e. it translocates in the xylem and phloem and thus offers “shoot to root” protection of the crop leaving pests nowhere to hide. It acts by inhibiting lipid biosynthesis, acting mainly after ingestion but also on contact, affecting eggs and larvae, with additional effects on adult fecundity. After penetrating the leaves, it is de-esterified and transported within both the xylem and phloem. It also possesses translaminar activity. It is highly effective in controlling the sucking pests including white flies, mealy bugs, root aphids and selected thrips in vegetables, cotton, soybeans and fruits.

Polysaccharides are major classes of biomolecules. They are biological polymers consisting of long chains of carbohydrate molecules, composed of several smaller units -monosaccharides. Polysaccharides contain more than ten monosaccharide units, whereas oligosaccharides contain three to ten monosaccharide units.

Their function in living organisms is usually either structure- or storage-related. Starch (a polymer of glucose) is used as a storage polysaccharide in plants, being found in the form of both amylose and the branched amylopectin. Cellulose and chitin are examples of structural polysaccharides. Cellulose is present in plant cell wall.

Modified polysaccharides are natural polysaccharides that have undergone chemical modifications to enhance their properties or create new functionalities, such as improved water solubility or enhanced biological activity. The structural modification of polysaccharides by chemical means can enhance their biological activity. The chemical modification of polysaccharides involves the use of chemical methods to modify some special structures or functional groups of polysaccharides on the main chains or side chains. Some of the chemical methods of modification include sulfation, phosphorylation, acetylation, carboxymethylation etc.
Studies have shown that the modified polysaccharide has various biological activities, such as antioxidant, antitumor, immune regulation, antiviral, antibacterial and anticoagulant functions in vitro.

In an embodiment, the polysaccharides or modified polysaccharides form an essential component of the insecticidal composition of the present invention. They play a dual role in growth stimulation, yield enhancement and as an adjuvant (thickener).

Polysaccharides or modified polysaccharides play a crucial role as formulation adjuvants in agrochemicals. They contribute in enhancing their efficacy and safety by enabling slow release, improving water-holding capacity, and reducing environmental impact.

Polysaccharides or modified polysaccharides can also play a significant role in enhancing crop growth and productivity through various biological and physiological mechanisms, as detailed below.

1. Improved Soil Health
Soil Structure and Water Retention: Polysaccharides secreted by soil microbes act as natural soil conditioners. They bind soil particles into aggregates, improving porosity, water retention, and aeration. This creates a favorable environment for root development of certain crops.
Enhanced Microbial Activity: Polysaccharides serve as carbon sources for beneficial soil microbes, promoting microbial activity that enhances nutrient cycling and availability.
2. Plant Growth Promotion
Bio stimulant Effect: Polysaccharides enhance plant metabolic activities, including photosynthesis, respiration, and enzyme production, leading to better growth and development of cotton plants.
Stress Tolerance: They help plants cope with abiotic stresses like drought, salinity, and heat by regulating osmotic balance and stabilizing cellular structures.

3. Nutrient Uptake and Utilization
Chelation of Nutrients: Polysaccharides can bind with essential nutrients (e.g., calcium, iron, and magnesium), making them more bioavailable to plants.
Improved Root Growth: By enhancing root elongation and branching, they facilitate better nutrient and water uptake.

4. Resistance to Pathogens
Induced Systemic Resistance (ISR): Some polysaccharides, like chitosan, stimulate the plant’s natural defense mechanisms, making plants more resistant to bacterial, viral and fungal diseases.

5. Growth Regulation
Hormonal Interaction: Polysaccharides influence the production of growth hormones like auxins and gibberellins, which promote cell division, elongation, and overall plant vigor.
Seed Germination: Polysaccharides improve seed hydration and enzyme activation, enhancing germination rates and seedling vigor.

In an embodiment, the insecticidal composition of the present invention comprises polysaccharide selected from the group comprising starch, cellulose, chitin, chitosan, arabinoxylans, pectins and combinations thereof.

In an embodiment, modified polysaccharides is selected from the group comprising modified forms of cellulose, starch, chitin, and chitosan, guar gum, sesbania gum, Acacia gum, karaya gum, mixture of agar and guar gum and combinations thereof.
The Applicant submits that present invention does not use any biological material originating in India. The polysaccharides or modified polysaccharides used in the instant invention is procured from Chongqing Shining Fine Chemicals Co., Ltd, China.

In an embodiment, the present invention provides a pesticidal composition comprising a) Flonicamid b) Spirotetramat and c) polysaccharides or modified polysaccharides along with at least one agriculturally acceptable excipient.

In an embodiment, the present invention provides an insecticidal composition comprising said combination that possesses an enhanced insecticidal efficacy at reduced cumulative dosage as compared to combined dosage of the solo active ingredient compounds when applied individually.

A combination of pesticidal actives may be applied sequentially or as tank mix or as ready -to- use composition or formulation. While sequential application increases the chemical load in the soil and hence is not an environmentally friendly approach, a tank mix is an unscientific practice adopted by the farmers which usually results in a heterogenous mixture. It has applicator and inhalation hazards besides other disadvantages like sedimentation issues, higher transportation costs and are environmentally less safe.

A ready-to-use (RTU) composition or formulation results in a homogenous composition, is easy to handle with lower chances of sedimentation, application hazard, phytotoxicity and are environmentally safer.

Thus, in an embodiment, the present invention provides a ready-to-use insecticidal composition comprising a) Flonicamid b) Spirotetramat and c) polysaccharides or modified polysaccharides along with at least one agriculturally acceptable excipient.

In a preferred embodiment, the present invention provides an insecticidal composition comprising a) Flonicamid in an amount ranging from 5-25% w/w b) Spirotetramat in an amount ranging from 5-25% w/w and c) polysaccharides or modified polysaccharides in an amount ranging from 0.2-2% w/w along with at least one agriculturally acceptable excipient.
In a more preferred embodiment, the present invention provides an insecticidal composition comprising Flonicamid in an amount of 18% w/w, Spirotetramat in an amount of 13.5% w/w, polysaccharides or modified polysaccharides in an amount of 2% w/w along with at least one agriculturally acceptable excipient.

In a further embodiment, the insecticidal composition of the present invention comprising said active ingredients along with polysaccharides or modified polysaccharides when combined in specific weight percentage, exhibits synergistic effect in control of sucking pests, particularly whitefly, jassids, aphids, thrips in agricultural crops, particularly in cotton.

In an embodiment, the insecticidal composition of the present invention is suitable for applying to a variety of agricultural crops including but not limited to cotton, cucurbits, okra, brinjal, chili, tomato, ornamental and horticultural crops.

In an embodiment, the insecticidal composition of the present invention further comprises at least one agriculturally acceptable excipient.

In an embodiment, said at least one agriculturally acceptable excipient is selected from the group comprising an anti-freezing agent, a wetting-cum-dispersing agent, an antifoaming agent, an antimicrobial agent and a thickening agent.

In a preferred embodiment, the insecticidal composition of the present invention is formulated as a suspension concentrate comprising Flonicamid active, Spirotetramat active, polysaccharides or modified polysaccharides, at least an anti-freezing agent, at least a wetting-cum-dispersing agent, at least an anti-foaming agent and at least an antimicrobial agent and at least a thickening agent.
In an embodiment, said anti-freezing agent is selected from the group comprising monoethylene glycol, diethylene glycol, polypropylene glycol, ethylene alcohols, polyethylene glycols, glycerine, epichlorohydrin, urea, glycerol, sorbitol or a mixture thereof. In a preferred embodiment, the antifreeze agent is polypropylene glycol. In an embodiment, the antifreeze agent is present in an amount ranging from 5-12% w/w.

In an embodiment, the wetting-cum-dispersing agent is selected from the group consisting of alkyl phenol Ethoxylate, salts of alkyl naphthyl sulphonate, salts of alkyl aryl sulphonate, derivative of sulfonated fatty alcohol, blend of aromatic sulphonic acid sodium salt, blend of alkyl aryl sulphonic acid sodium salt, sodium -N-Methyl-N-Oleyl taurate. In a preferred embodiment, said wetting-cum-dispersing agent is alkyl phenol ethylene oxide ammonium sulphonates. In an embodiment, the wetting-cum-dispersing agent is present in an amount ranging from 2.0-8.0 %w/w.

In an embodiment, the antifoaming agent is selected from the group consisting of silicone emulsions, long-chain alcohols, fatty acids, organic fluorine compounds and combinations thereof. In a preferred embodiment, the antifoaming agent is silicon based. In a preferred embodiment, the antifoaming agent is present in an amount ranging from 0.1-2.0% w/w.

In an embodiment, the antimicrobial agent is selected from the group comprising Clavulanic acid, Avibactum, stigmasterol, formaldehyde, organic compounds including Benzisothiazolinone. In a preferred embodiment, the antimicrobial agent is 1,2- Benzisothiazolin-3-one. In an embodiment, the anti-microbial agent is present in an amount ranging from 0.1-1.0 % w/w.

In an embodiment, other well-known thickening agents may be used in the present invention. Such a thickening agent may be selected from the group comprising polyacrylamide, polyethylene polymers, polysaccharides (long-chain 15 sugars), vegetable oils, xanthan gum. In a preferred embodiment, the thickening agent is polysaccharides or modified polysaccharides.

In the most preferred embodiment, the presently claimed insecticidal composition comprises Flonicamid in an amount of 18% w/w, Spirotetramat in an amount of 13.5% w/w, propylene glycol (anti-freeze agent) in an amount of 7% w/w, alkylphenol ethylene oxide ammonium sulphonates (wetting-cum dispersing agent) in an amount of 3% w/w, polyalkyl aryl ethyele oxide phosphate ester (emulsifier) in an amount of 2%w/w, Polydimethyl siloxane (anti-foaming agent) in an amount of 0.5% w/w, 1,2- Benzisothiazolin-3-one (anti-microbial agent) in an amount of 0.2% w/w, polysaccharides or modified polysaccharides (thickener and bio stimulant) in an amount of 2% w/w and Q.S. demineralised water.
In an embodiment, the insecticidal composition of the present invention is formulated in a form selected from the group comprising water-soluble concentrates (SL), emulsifiable concentrates (EC), emulsions (EW), micro-emulsions (ME), Suspension concentrates (SC), oil-based suspension concentrates (OD), flowable suspensions (FS), water-dispersible granules (WG), water-soluble granules (SG), wettable powders (WP), water soluble powders (SP), granules (GR), encapsulated granules (CG), fine granules (FG), macrogranules (GG), dry flowables (DF), aqueous suspo-emulsions (SE), capsule suspensions (CS) and microgranules (MG). In a preferred embodiment, the insecticidal composition is in the form of suspension concentrates (SC), water-dispersible granule (WDG) and wettable powder (WP).

In a preferred embodiment, the insecticidal composition of the present invention is formulated as a suspension concentrate (SC), wherein said suspension concentrate comprises Flonicamid active, Spirotetramat active, polysaccharides or modified polysaccharide, anti-freezing agent, a wetting -cum-dispersing agent, an antifoaming agent, a thickening agent and an antimicrobial.

A suspension concentrate offers many advantages over other types of formulations. It is water-based and provides good safety and user convenience. It is suitable for many active ingredients with low water solubility. It has built- in adjuvants that can be used for bio enhancement. Absence of dusts and flammable liquids in this formulation type reduces the application hazards.

EXAMPLES:
The examples below are given solely for the purpose of illustration and are not to be construed as limitations of the present invention, as many variations thereof are possible without departing from the spirit and scope of the invention.

Example 1: Preparation of insecticidal composition as a suspension concentrate

In a preferred embodiment, the chemical constitution of the present insecticidal composition is depicted below in Table 1.
Table 1: Preferred composition
S.no. Constituent Weight Percentage w/w
1. Flonicamid (Basis of 100%) 18.0
2. Spirotetramat (Basis of 100%) 13.5
3. Polysaccharide or Modified Polysaccharides (Thickener and Bio Stimulant) 2
4. Antifreeze agent (Propylene glycol) 7.0
5. Wetting-cum-dispersing agent (alkyl phenol ethylene oxide ammonium sulphonates) 3.0
6. Anti foaming agent (Polydimethyl siloxane) 0.5
7. Anti-bacterial (1,2-Benzisothiazolin-3-one) 0.2
8. Demineralised water Q.S
Total

Preparation method of Suspension concentrate (SC):
Step 1: Required amount of demineralised water was taken in a pre-mixing vessel followed by addition of anti- freezing agent (Propylene Glycol), dispersing agent (Alkyl phenol ethylene oxide ammonium sulphonates), wetting agent (Poly alkylaryl ethylene oxide phosphate ester) under continuous stirring and required amount of silicon based anti-foaming agent (Polydimethyl siloxane) was added to reduce foaming.
Step 2: Spirotetramat technical was gradually added to the vessel followed by Flonicamid technical under continuous stirring.
Step 3: The material was passed through a Dyno Mill to reduce the particle size. The mean particle size value targeted was D (90) <10 µ (microns)
Step 4: The material was transferred in post mixing blender and balance quantity of antifoaming agent (Polydimethyl siloxane), antimicrobial (1,2-Benzisothiazolin-3-one) and thickener and stimulant (polysaccharides or modified polysaccharides) was added to the mix at low RPM to get a homogeneous suspension.
Step 5: After mixing all the contents till homogeneous, quality parameters (such as wettability, suspensibility, etc.) were checked. If tested ok desired packing or transfer to storage vessel was carried out.

Example 2: Field evaluation of the bio efficacy of the present Insecticidal composition
The presently disclosed insecticidal combination was tested for its bioefficacy against White fly (Bemisia tabaco) and jassids (Amrasca biguttula) in a local variety of cotton. The plants were aligned in a plot size of 50 sq. meter in a randomized block design. The trial was carried out in Aurangabad, Maharashtra during the kharif season 2024-2025 at a temp 25? and 74% relative humidity and windspeed of roughly 3.7 m/s. A total of 12 treatments were carried out in three replications. The application of the insecticidal trimix was foliar (over the top).

Measured quantity of the chemical was added to required volume of water @ 375 lit. /ha for spray. The spray tank was filled with half the quantity of clean required volume of water and then the measured chemical (according to the dose) was added followed by the rest half quantity of water. The solution was stirred well before application while ensuring thorough coverage.

Table 2 below provides the treatment details in cotton.
Table 2: Treatment Details.
Treatment Dose a.i./ha (gm) Dose Formulation (gm/ml/ha)
Untreated
Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC (RTU) 54+ 40.5 + 6 300 ml
Flonicamid 18% + Spirotetramat 13.5% SC (RTU w/o polys) 54+ 40.5 300 ml
Flonicamid 50% WG + Spirotetramat 15.31 OD (Tank Mix) 54 + 40.5 108 gm + 300 ml
Flonicamid 50% WG + Spirotetramat 15.31 OD + Polysaccharides mixtures 2% SC (Tank Mix) 54 + 40.5 +6 108 gm + 300 ml + 300 ml
Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC (RTU) 72 + 54 +8 400 ml
Flonicamid 18% + Spirotetramat 13.5% SC (RTU w/o polys) 72 + 54 400 ml
Flonicamid 50% WG + Spirotetramat 15.31 OD + Polysaccharides mixtures 2% SC (Tank Mix) 72 + 54 + 8 144 + 350 + 400ml
Flonicamid 50% WG + Spirotetramat 15.31 OD (Tank Mix) 72 + 54 144 + 350ml
Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC (RTU) 75 + 60 +10 416 ml
Flonicamid 18% + Spirotetramat 13.5% SC (RTU w/o polys) 75 + 60 416 ml
Flonicamid 50% WG + Spirotetramat 15.31 OD + Polysaccharides mixtures 2% SC (Tank Mix) 75 + 60 +10 150 + 60 + 500 ml
Flonicamid 50% WG + Spirotetramat 15.31 OD (Tank Mix) 75 + 60 150 + 60
Flonicamid 50 WG (UPL)- solo application 75 150g
Spirotetramat 15.31 OD (BAYER)- solo application 60 400 ml
Polysaccharides mixtures 2% SC)- solo application 10 500 ml
Note: RTU: Ready to use
RTU w/o polys: Ready to use without polysaccharides

Each plot measured 5 × 5 meters, with cotton sown at a standard spacing. Recommended agronomic practices were followed throughout the study.

Evaluation of Bio efficacy:
The bio-efficacy evaluation involved counting the number of live Jassids and Whiteflies on 10 tagged plants before application (pre-count) and at intervals of 1, 3, 5, 7 and 10 days after application (DAA), as well as at 5-day intervals thereafter until re-infestation. Observations included the count of live adults and nymphs from 30 leaves across the tagged plants. The percentage control of the live population was calculated using the formula based on the number of live sucking pests (Jassids and Whiteflies).

Percent Control=(C-T)/C×100

Where C is No. of live Jassids/Whitefly population in control plot
T is No. of live Jassids/Whitefly population in treated plot

Data Collection:
Pest population assessment pre-treatment and post-treatment pest counts both Jassids and Whitefly were recorded at 3, 7, and 10 days after treatment (DAT) from five randomly selected plants per plot. Crop parameters observations on crop vigor and leaf damage were taken to correlate pest suppression with plant health. The final seed cotton yield data was recorded to assess treatment impact.

Statistical Analysis:
Analysis of Variance (ANOVA):
ANOVA was performed to compare the mean pest population, crop health parameters, and yield data across treatments. Significant differences among treatments were identified using the least significant difference (LSD) test at a 5% significance level (Gomez & Gomez, 1984).

Colby’s Method for Synergism/Antagonism:
To determine interaction effects, the observed mortality rates of Jassids and Whitefly in two-three-way combinations were compared with expected mortality rates using Colby's formula (Colby, 1967).

The synergistic action expected for a given combination of two-way active components can be calculated as follows:

E=A+B-AB/100
The synergistic action expected for a given combination of three-way active components can be calculated as follows:

E=(A+B+C)-((AB+BC+CA))/100+ABC/10000

where ?? is the expected mortality, and A, B and C are the observed mortalities of individual components.

Synergistic effects were indicated if the observed mortality was greater than the expected mortality, while antagonistic effects were noted if the observed mortality was lower.

Validation and Interpretation:
Results from ANOVA provided insights into treatment effectiveness, while Colby’s method elucidated the interactions among insecticide components. The combined results guided the evaluation of the novel formulation's performance in pest suppression and yield enhancement.

The average values for different mixtures of insecticides on cotton were assessed against jassids and whitefly, focusing on key parameters such as the number of squares per plant, number of flowers per plant, number of balls per plant, and total yield (qt/acre). The mixtures tested included doses (gm a.i./ha) of 54+40.5+6 a.i./ha (gm). Results indicated a progressive improvement in plant productivity and yield with increasing doses, demonstrating the efficacy of the mixtures in controlling Jassids and promoting overall crop health and productivity. Detailed analysis highlights the optimal dose for maximum yield in table no. 5.

Table 3: Mean value of various mixtures on Cotton against jassids and whitefly
Treatment Dose a.i./ha (gm) Dose Formulation (gm/ml/ha) Mean Value
No. of square/Plants No. of Flower/Plants No. of Bolls/Plants Total Yield (qt/acre)
Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC (RTU) 54+ 40.5 + 6 300 ml 16.07 19.23 23.81 23.81
Flonicamid 18% + Spirotetramat 13.5% SC (RTU w/o polys) 54+ 40.5 300 ml 8.93 7.69 4.76 4.76
Flonicamid 50% WG + Spirotetramat 15.31 OD (Tank Mix) 54 + 40.5 108 gm + 300 ml 12.50 15.38 23.81 23.81
Flonicamid 50% WG + Spirotetramat 15.31 OD + Polysaccharides mixtures 2% SC (Tank Mix) 54 + 40.5 +6 108 gm + 300 ml + 300 ml 10.71 11.54 14.29 14.29
Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC (RTU) 72 + 54 +8 400 ml 17.86 17.31 14.29 14.29
Flonicamid 18% + Spirotetramat 13.5% SC (RTU w/o polys) 72 + 54 400 ml 9.82 11.54 14.29 14.29
Flonicamid 50% WG + Spirotetramat 15.31 OD + Polysaccharides mixtures 2% SC (Tank Mix) 72 + 54 + 8 144 + 350 + 400 20.54 19.23 24.76 24.76
Flonicamid 50% WG + Spirotetramat 15.31 OD (Tank Mix) 72 + 54 144 + 350 11.61 11.54 9.52 9.52
Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC (RTU) 75 + 60 +10 416 ml 17.86 15.38 23.81 23.81
Flonicamid 18% + Spirotetramat 13.5% SC (RTU w/o polys) 75 + 60 416 ml 10.71 3.85 7.62 7.62
Flonicamid 50% WG + Spirotetramat 15.31 OD + Polysaccharides mixtures 2% SC (Tank Mix) 75 + 60 +10 150 + 60 + 500 ml 18.75 14.23 14.29 14.29
Flonicamid 50% WG + Spirotetramat 15.31 OD (Tank Mix) 75 + 60 150 + 60 14.29 7.69 10.44 10.44
Flonicamid 50 WG (UPL) -solo 75 150 3.57 3.85 3.81 3.81
Spirotetramat 15.31 OD (BAYER)- solo 60 400 7.14 7.69 3.81 3.81
Polysaccharides mixtures 2% SC- solo 10 500 ml 3.57 4.62 7.62 7.62
SE (m) 0.14 0.15 0.12 0.29
C.D. 5% 0.43 0.44 0.37 0.84
* Normalisations: Arc sin transformed value used: SEM; Standard error of means; CD@5% level of significance: Critical Difference
Note : RTU : Ready to use
RTU w/o polys: Ready to use without polysaccharides
Table 4: Synergistic effect of Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC on cotton crop for Jassids
S. No. Treatment Dose a.i./ha (gm) Dose Formulation (gm/ml/ha) % reduction against control (Observed-Jassids) % reduction expected (Colby’s formula)
1 Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC (RTU) 54+ 40.5 + 6 300 ml 91.90 89.57
2 Flonicamid 18% + Spirotetramat 13.5% SC (RTU w/o polys) 54+ 40.5 300 ml 83.30 87.91
3 Flonicamid 50% WG + Spirotetramat 15.31 OD (Tank Mix) 54 + 40.5 108 gm + 300 ml 72.16 87.91
4 Flonicamid 50% WG + Spirotetramat 15.31 OD + Polysaccharides mixtures 2% SC (Tank Mix) 54 + 40.5 +6 108 gm + 300 ml + 300 ml 66.59 89.57
5 Flonicamid 50 WG (UPL) 75 150 63.81 -
6 Spirotetramat 15.31 OD (BAYER) 60 400 66.59 -
7 Polysaccharides mixtures 2% SC 10 500 ml 13.70 -

Table 5: Synergistic effect of Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC on cotton crop for Jassids
S. No. Treatment Dose a.i./ha (gm) Dose Formulation (gm/ml/ha) % reduction against control (Observed-Jassids) % reduction expected (Colby’s formula)
1 Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC (RTU) 72 + 54 +8 400 ml 91.65 89.57
2 Flonicamid 18% + Spirotetramat 13.5% SC
(RTU w/o polys) 72 + 54 400 ml 88.86 87.91
3 Flonicamid 50% WG + Spirotetramat 15.31 OD + Polysaccharides mixtures 2% SC (Tank Mix) 72 + 54 + 8 144 + 350 + 400 80.51 89.57
4 Flonicamid 50% WG + Spirotetramat 15.31 OD (Tank Mix) 72 + 54 144 + 350 72.16 87.91
5 Flonicamid 50 WG (UPL) 75 150 63.81 -
6 Spirotetramat 15.31 OD (BAYER) 60 400 66.59 -
7 Polysaccharides mixtures 2% SC 10 500 ml 13.70 -

Table 6: Synergistic effect of Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC on cotton crop for Jassids
S. No. Treatment Dose a.i./ha (gm) Dose Formulation (gm/ml/ha) % reduction against control (Observed-Jassids) % reduction expected (Colby’s formula)
1 Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC (RTU) 75 + 60 +10 416 ml 94.43 89.57
2 Flonicamid 18% + Spirotetramat 13.5% SC (RTU w/o polys) 75 + 60 416 ml 88.86 87.91
3 Flonicamid 50% WG + Spirotetramat 15.31 OD + Polysaccharides mixtures 2% SC (Tank Mix) 75 + 60 +10 150 + 60 + 500 ml 80.51 89.57
4 Flonicamid 50% WG + Spirotetramat 15.31 OD (Tank Mix) 75 + 60 150 + 60 72.16 87.91
5 Flonicamid 50 WG (UPL) 75 150 63.81 -
6 Spirotetramat 15.31 OD (BAYER) 60 400 66.59 -
7 Polysaccharides mixtures 2% SC 10 500 ml 13.70 -

Calculation by Colbys method:
The synergistic action expected for a given combination of two-way active components can be calculated for jassids:

Expected (E)=63.81+66.59-(63.81*66.59)/100

Expected (E)=87.91

The synergistic action expected for a given combination of three-way active components can be calculated for jassids:

Expected (E)=(63.81+66.59+13.70)-((63.81*66.59+66.59*13.70+13.70*63.81))/100+(63.81*66.59*13.70)/10000

Expected (E)=89.57

Table 7: Synergistic effect of Flonicamid 18% + Spirotetramat 13.5% SC + polysaccharides mixtures 2% SC on cotton crop for whitefly
S. No. Treatment Dose a.i./ha (gm) Dose Formulation (gm/ml/ha) % reduction against control (Observed-Whitefly) % reduction expected (Colby’s formula)
1 Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC (RTU) 54+ 40.5 + 6 300 ml 88.92 88.72
2 Flonicamid 18% + Spirotetramat 13.5% SC (RTU w/o polys) 54+ 40.5 300 ml 82.86 87.26
3 Flonicamid 50% WG + Spirotetramat 15.31 OD (Tank Mix) 54 + 40.5 108 gm + 300 ml 71.43 87.26
4 Flonicamid 50% WG + Spirotetramat 15.31 OD + Polysaccharides mixtures 2% SC (Tank Mix) 54 + 40.5 +6 108 gm + 300 ml + 300 ml 65.71 88.72
5 Flonicamid 50 WG (UPL) 75 150 62.86 -
6 Spirotetramat 15.31 OD (BAYER) 60 400 65.71 -
7 Polysaccharides mixtures 2% SC 10 500 ml 11.43 -

Table 8: Synergistic effect of Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC on cotton crop for Whitefly
S. No. Treatment Dose a.i./ha (gm) Dose Formulation (gm/ml/ha) % reduction against control (Observed-Whitefly) % reduction expected (Colby’s formula)
1 Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC (RTU) 72 + 54 +8 400 ml 91.43 88.72
2 Flonicamid 18% + Spirotetramat 13.5% SC (RTU w/o polys) 72 + 54 400 ml 88.57 87.26
3 Flonicamid 50% WG + Spirotetramat 15.31 OD + Polysaccharides mixtures 2% SC (Tank Mix) 72 + 54 + 8 144 + 350 + 400 80.00 88.72
4 Flonicamid 50% WG + Spirotetramat 15.31 OD (Tank Mix) 72 + 54 144 + 350 71.43 87.26
5 Flonicamid 50 WG (UPL) 75 150 62.86 -
6 Spirotetramat 15.31 OD (BAYER) 60 400 65.71 -
7 Polysaccharides mixtures 2% SC 10 500 ml 11.43 -

Table 9: Synergistic effect of Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC on cotton crop for Whitefly
S. No. Treatment Dose a.i./ha (gm) Dose Formulation (gm/ml/ha) % reduction against control (Observed-Whitefly) % reduction expected (Colby’s formula)
1 Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC(RTU) 75 + 60 +10 416 ml 94.29 88.72
2 Flonicamid 18% + Spirotetramat 13.5% SC (RTU w/o polys) 75 + 60 416 ml 88.57 87.26
3 Flonicamid 50% WG + Spirotetramat 15.31 OD + Polysaccharides mixtures 2% SC (Tank Mix) 75 + 60 +10 150 + 60 + 500 ml 80.00 88.72
4 Flonicamid 50% WG + Spirotetramat 15.31 OD (Tank Mix) 75 + 60 150 + 60 71.43 87.26
5 Flonicamid 50 WG (UPL) 75 150 62.86 -
6 Spirotetramat 15.31 OD (BAYER) 60 400 65.71 -
7 Polysaccharides mixtures 2% SC 10 500 ml 11.43 -

Calculation:
The synergistic action expected for a given combination of two-way active components can be calculated for jassids:

Expected (E)=62.86+65.71-(60.86*65.71)/100

Expected (E)=87.26

The synergistic action expected for a given combination of three-way active components can be calculated for jassids:

Expected (E)=(62.86+65.71+11.43)-((62.86*65.71+65.71*11.43+11.43*62.86))/100+(62.86*65.71*11.43)/10000

Expected (E)=88.72

Conclusion
The efficacy achieved through application of Flonicamid 18%, Spirotetramat 13.5% SC, and polysaccharides mixtures 2% SC against Jassids and whiteflies on cotton was evaluated using ANOVA and Colby’s method to assess pest control and synergistic effects. The study tested the three-way combination (Flonicamid+ Spirotetramat + modified polysaccharides) at the dosage of 54+40.5+6 a.i./ha (g), alongside higher doses, to determine their effectiveness in reducing pest infestations and enhancing crop yields.
ANOVA results revealed significant differences in pest reduction and yield improvements across the tested doses. The optimized dose of 54+40.5+6 a.i./ha (gm) demonstrated superior control over Jassids and whitefly infestations (measured on five leaves per plant). This dose consistently achieved the best outcomes in terms of pest mortality and crop yield, with statistically significant improvements compared to other tested dosage or individual components.

Colby’s method was employed to evaluate the potential synergistic interactions between the formulation components. The analysis confirmed synergy, as the observed pest mortality rates exceeded the expected additive effects at most of the dosages tested.

The synergy was most pronounced at the dose (54+40.5+6 a.i./ha), where the active combination exhibited amplified efficacy and yield enhancement. The results underscore the robustness of the three-way combination of Flonicamid, Spirotetramat and modified polysaccharides as a pest management solution for cotton.

The observed synergy not only enhances pest control but also ensures higher yields, making the formulation a valuable component of integrated pest management (IPM) strategies. By improving both pest control and productivity, this combination provides significant economic and agricultural benefits to cotton growers.

Example 3: Evaluation of Phytotoxicity:
Observations were recorded at 3, 7, and 10 days after application to evaluate potential phytotoxic effects. The parameters assessed included leaf injury on tips and surfaces, necrosis, vein clearing, epinasty, hyponasty, and wilting. Each parameter was rated using a standard phytotoxicity scale ranging from 0 to 10, where 0 indicated no damage and 10 represented severe damage. This systematic approach ensured consistent and accurate assessment of the treatment's impact on plant health, allowing for a comprehensive evaluation of its safety and suitability for use in cotton cultivation.

The results of the observations are tabulated below:
Table 10: Phytotoxicity symptoms scoring and rating for leaf injury on tip/surface
Leaf injury on tips/surface 0% 1-10% 11-20% 21-30% 31-40% 41-50% 51-60% 61-70% 71-80% 81-90% 91-100%
Rating 0 1 2 3 4 5 6 7 8 9 10

Table 11: Phytotoxicity effect of composition Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC on cotton crop for Jassids and Whitefly
Treatment Dose a.i./ha (gm) Dose Formulation (gm/ml/ha) 1DAA 3 DAA 5
DAA 7 DAA 10
DAA
Untreated 0 0 0 0 0
Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC (RTU) 54+ 40.5 + 6 300 ml 0 0 0 0 0
Flonicamid 18% + Spirotetramat 13.5% SC (RTU w/o polys) 54+ 40.5 300 ml 0 0 0 0 0
Flonicamid 50% WG + Spirotetramat 15.31 OD (Tank Mix) 54 + 40.5 108 gm + 300 ml 0 0 0 0 0
Flonicamid 50% WG + Spirotetramat 15.31 OD + Polysaccharides mixtures 2% SC (Tank Mix) 54 + 40.5 +6 108 gm + 300 ml + 300 ml 0 0 0 0 0
Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC (RTU) 72 + 54 +8 400 ml 0 0 0 0 0
Flonicamid 18% + Spirotetramat 13.5% SC (RTU w/o polys) 72 + 54 400 ml 0 0 0 0 0
Flonicamid 50% WG + Spirotetramat 15.31 OD + Polysaccharides mixtures 2% SC (Tank Mix) 72 + 54 + 8 144 + 350 + 400 0 0 0 0 0
Flonicamid 50% WG + Spirotetramat 15.31 OD (Tank Mix) 72 + 54 144 + 350 0 0 0 0 0
Flonicamid 18% + Spirotetramat 13.5% SC + Polysaccharides mixtures 2% SC (RTU) 75 + 60 +10 416 ml 0 0 0 0 0
Flonicamid 18% + Spirotetramat 13.5% SC (RTU w/o polys) 75 + 60 416 ml 0 0 0 0 0
Flonicamid 50% WG + Spirotetramat 15.31 OD + Polysaccharides mixtures 2% SC (Tank Mix) 75 + 60 +10 150 + 60 + 500 ml 0 0 0 0 0
Flonicamid 50% WG + Spirotetramat 15.31 OD (Tank Mix) 75 + 60 150 + 60 0 0 0 0 0
Flonicamid 50 WG (UPL) 75 150 0 0 0 0 0
Spirotetramat 15.31 OD (BAYER) 60 400 0 0 0 0 0
Polysaccharides mixtures 2% SC 10 500 ml 0 0 0 0 0

Results and Discussion
Phytotoxicity symptoms of jassids and whitefly, including leaf injury on tips and surfaces, were systematically scored and rated using a standardized scale ranging from 0 to 10, where 0 indicated no visible symptoms and 10 represented severe damage. During the evaluation at 1, 3, 5, 7, and 10 days after application, no phytotoxicity symptoms such as necrosis, vein clearing, epinasty, hyponasty, or wilting were observed on the treated plants. The absence of leaf injury and other adverse effects confirmed the safety of the tested insecticide combination, indicating its suitability for use without compromising the health and vigor of the cotton crop in table no. 4 against jassids and whitefly.
Therefore, the present inventors arrived at the conclusion that the combination of Flonicamid 18%, Spirotetramat 13.5% SC and polysaccharides or modified polysaccharides mixtures 2% SC is highly effective in controlling Jassids and whiteflies in cotton. The optimal dose of 54+40.5+6 a.i./ha (gm) consistently delivered the best results, offering enhanced efficacy, synergistic action without any noticeable phytotoxicity thus delivering a promising sustainable and economical approach to pest management.
, Claims:We Claim:
1. An insecticidal composition comprising:
a) Flonicamid in an amount ranging from 5-25 % w/w.
b) Spirotetramat in an amount ranging from 5-25 % w/w and
c) Polysaccharides or modified polysaccharides in an amount ranging from 0.2-2% w/w and
d) at least one agriculturally acceptable excipient

2. The insecticidal composition as claimed in claim 1, wherein said polysaccharide is selected from the group comprising from the group comprising starch, cellulose, chitin, chitosan, arabinoxylans, pectin and combinations thereof.

3. The insecticidal composition as claimed in claim 1, wherein said modified polysaccharides is selected from the group comprising modified forms of cellulose, starch, chitin, and chitosan, guar gum, sesbania gum, Acacia gum, karaya gum , mixture of agar and guar gum and combinations thereof.

4. The insecticidal composition as claimed in claim 1, wherein said at least one agriculturally acceptable excipient comprises an anti-freezing agent, a wetting-cum- dispersing agent, an antifoaming agent and an antimicrobial agent.

5. The insecticidal composition as claimed in claim 4, wherein the anti-freeze agent is present in an amount ranging from 5-12 %w/w, wetting-cum-dispersing agent is present in an amount ranging from 2-8% w/w, antifoaming agent is present in an amount ranging from 0.1-2% w/w and antimicrobial agent is present in an amount ranging from 0.1-1% w/w.

6. The insecticidal composition as claimed in claim 4, wherein anti-freeze agent is selected from the group comprising monoethylene glycol, diethylene glycol, polypropylene glycol, ethylene alcohols, polyethylene glycols, glycerine, epichlorohydrin, urea, glycerol, sorbitol, or a mixture thereof.

7. The insecticidal composition as claimed in claim 4, wherein the wetting-cum-dispersing agent is selected from the group comprising alkyl Phenol Ethoxylate, salts of alkyl naphthyl sulphonate, salts of alkyl aryl sulphonate, derivative of sulfonated fatty alcohol, blend of aromatic sulphonic acid sodium salt, Blend of alkyl aryl sulphonic acid sodium salt and sodium -N-Methyl-N-Oleyl taurate.

8. The insecticidal composition as claimed in claim 4, wherein the anti-foaming agent is selected from the group comprising silicone emulsions, long-chain alcohols, fatty acids, organic fluorine compounds and combinations thereof.

9. The insecticidal composition as claimed in claim 4, wherein the antimicrobial agent is selected from the group comprising Clavulanic acid, Avibactum, stigmasterol, formaldehyde and Benzisothiazolinone.

10. The insecticidal composition as claimed in claim 1, wherein said insecticidal composition is formulated in a form selected from the group comprising water-soluble concentrates (SL), emulsifiable concentrates (EC), emulsions (EW), micro-emulsions (ME), Suspension concentrates (SC), oil-dispersion (OD), flowable suspensions (FS), water-dispersible granules (WG), water-soluble granules (SG), wettable powders (WP), water soluble powders (SP), dry flowables (DF), granules (GR), encapsulated granules (CG), fine granules (FG), macrogranules (GG), aqueous Suspo-emulsions (SE), capsule suspensions (CS) and microgranules (MG).

11. The insecticidal composition as claimed in claim 1, wherein said insecticidal composition is preferably formulated in the form of a suspension concentrate.

12. The insecticidal composition as claimed in claim 1, wherein Flonicamid is present in an amount of 18%w/w, Spirotetramat is present in an amount of 13.5%w/w, polysaccharide or modified polysaccharide is present in an amount of 2% w/w, an antifreeze agent in an amount of 7% w/w, wetting -cum-dispersing agent in an amount of 3% w/w, anti-foaming agent in an amount of 0.5% w/w and anti-microbial agent in an amount of 0.2% w/w of the insecticidal composition.

13. A process for preparing an insecticidal composition as claimed in claims 1-12 in the form of a suspension concentrate, said process comprising:
a) adding a required amount of demineralized water to a pre-mixing vessel, followed by the addition of an anti-freezing agent (propylene glycol), a wetting-cum dispersing agent (namely alkyl phenol ethylene oxide ammonium sulphonates) under continuous stirring and adding silicone-based antifoaming agent (polydimethyl siloxane);
b) adding sequentially spirotetramat technical and flonicamid technical into the vessel under continuous stirring;
c) milling the mixture in a Dyno Mill to obtain a particle size D(90) is less than 10 micrometers;
d) transferring the milled material to a post-mixing blender and adding the remaining quantity of the antifoaming agent, polydimethyl siloxane, an antimicrobial agent, 1,2-benzisothiazolin-3-one, and a thickener, selected from polysaccharides or modified polysaccharides, at a low rotational speed to ensure a homogeneous suspension; and
e) homogenizing the mixture, assessing quality parameters including wettability and suspensibility, and testing, and proceeding to packaging or transferring the final product to a storage vessel.

14. The process as claimed in claim 13, wherein Flonicamid is present in an amount ranging from 5-25% w/w, Spirotetramat is present in an amount ranging from 5-25% w/w and modified polysaccharide is present in an amount ranging from 0.2-2% w/w.

15. A method of controlling or preventing insect pests on plants or propagation materials thereof, said method comprising applying an insecticidally effective amount of the insecticide composition as claimed in claims 1-12, on a plant or a part thereof or at the locus or on the surface of plant propagation material.