1. A herbal pesticide, herbal growth promoter and antimicrobial formulation comprising; a) seed oil of Azadiracta indica; b) leaves oil of Cymbopogon citratus; c) grain flour of Pennisetum typhoides; d) tween 80S; e) oleic acid; and f) water wherein this formulation is in a microemulsion form.

2. A herbal pesticide, herbal growth promoter and antimicrobial formulation comprising; a) seed oil of Azadiracta indica; b) leaves oil of Cymbopogon citratus; c) grain flour of Pennisetum typhoides; d) citric acid; e) tartaric acid; and f) sodium bicarbonate wherein this formulation is in a dispersible tablet form.

3. The herbal pesticide, herbal growth promoter and antimicrobial formulation as claimed in claim 1 wherein 25 ml of microemulsion formulation comprises 7.5 mg grain flour of Pennisetum typhoides, 4 ml seed oil of Azadiracta indica and 4 ml leaves oil of Cymbopogon citratus.

4. The herbal pesticide, herbal growth promoter and antimicrobial formulation as claimed in claim 1 wherein 25 ml of microemulsion comprises 3 ml of tween 80S and 3 ml of oleic acid as emulsifying agents, and 11 ml water.

5. The herbal pesticide, herbal growth promoter and antimicrobial formulation as claimed in claim 2 wherein a batch of 11 gm (11 tablets of 1 gm each) comprises 1 ml seed oil of Azadiractica indica, 1 ml leaves oil of Cybopogon citratus and 1 gm grain flour of Pennisetum typhoides.

6. The herbal pesticide, herbal growth promoter and antimicrobial formulation as claimed in claim 2 wherein a batch of 11 gm (11 tablets of 1 gm each) comprises 1.8 gm of citric acid, 2.7 gm of tartaric acid and 5.1 gm of sodium bicarbonate as effervescent.

7. The herbal pesticide, herbal growth promoter and antimicrobial formulation as claimed in claim 1 wherein microemulsion formulation is prepared by a process comprising steps of; a. Mixing of seed oil of Azadiractica indica and leaves oil of Cybopogon citratus in 1:1 (4 ml each). b. Adding of grain flour of Pennisetum typhoides (7.5 mg) in above mixture. Stirring well to form a homogenous mixture (mixture A). c. Mixing 3 ml tween 80S and 3 ml oleic acid (1:1) in a separate beaker. Stirring well to form the mixture B. d. Adding mixture B to mixture A on a sonicator. e. Sonicating the resultant mixture for 15 minutes. f. Adding 11 ml water and keeping the final mixture for 5 minutes followed by sonication to form a proper microemulsion.

8. The herbal pesticide, herbal growth promoter and antimicrobial formulation as claimed in claim 1 wherein dispersible tablet formulation is prepared by a process comprising steps of; a. Milling and mixing of 1.8 gm citric acid, 2.7 gm tartaric acid, 5.1 gm sodium bicarbonate and 1 gm of grain flour of Pennisetum typhoides in mortar and pestle. b. Preparing a wet mass of above powder mixture by using 1 ml seed oil of Azadiractica indica and 1 ml leaves oil of Cybopogon citratus as active ingredient and as solvent. Planetry mixer is used for wet mixing of the powders. c. Forcing the wet mass of step b through a sieve (20 mesh screen) to produce wet granules. d. Drying the wet granules in oven at 60 degree celcius temperature for 2 hrs. e. Compressing the dried granules of step d by tablet press machine at a pressure of 6 tonnes.

FIELD OF INVENTION

The purpose of this invention is to prevent the side effects of synthetic pesticides and to utilize the benefits of herbal pesticides over them. The present invention relates to herbal pesticide formulations. This invention is specifically related with herbal pesticidal formulations in microemulsion and tablet form. The present invention also provide a process for the preparation of these herbal pesticide formulations.

BACKGROUND OF INVENTION

Introduction

Insects are responsible for two major kinds of damage to growing crops. First is direct injury done to the plant by the feeding insect, which eats leaves or burrows in stems, fruit, or roots. There are hundreds of pest species of this type, both in larvae and adults, among orthopterans, homopterans, heteropterans, coleopterans, lepidopterans, and dipterans. The second type is indirect damage in which the insect itself does little or no harm but transmits a bacterial, viral, or fungal infection into a crop. Examples include the viral diseases of sugar beets and potatoes, carried from plant to plant by aphids. Generally, synthetic pesticides (insecticides) are used to remove these insects. Synthetic pesticides have harmful effects not only on flora and fauna but also on the environment. Many incidence of mortality of farmers due to toxic effects of synthetic pesticides have been reported. It is also reported that many farmers have committed suicide by consuming the synthetic pesticide. These types of incidence and harmful effects of synthetic pesticides can be prevented by using herbal pesticides which are environmental friendly and non-toxic to humans.

Pesticidal agents

Pests are organisms that might compete with or damage crop species. They reduce plant density, cause plant stunted growth and their death, cause lower production capacity, damage berries and in many other ways reduce the yield or quality of agricultural products. Agricultural pests are plants, animals and fungus that reduce crop yields. One of the most common ways to control or eliminate pests is to use poisons that kill pests, pesticides. They work by causing physical or biological harm to the pest organisms and may interfere with biological processes, such as photosynthesis, or cause damage to vital organs. Alternatively, some pesticides are indirectly applied or can be sprayed on a plant and later consumed by an insect.
Pesticides provide many substantial benefits for farmers and consumers by controlling insects and preventing diseases, as well as increasing crop yield and keeping cost down; however, these potent chemicals have also put our health in great danger. The side effects of synthetic pesticides (organophosphate, organochlorine, carbamates etc.) includes cancer, endocrine complications, infertility and sterility, brain damage, birth defects, respiratory disorders, organ failure, skin irritations etc.
Botanical pesticides or insecticides have a proven track record and long use. Public concern over use of synthetic pesticides is growing. So botanicals are now replacement for synthetics , the new environmental movement has provided a favourable environment for the rebirth of botanical insecticides.
Synthetic pesticides are very efficient in insect control but can be harmful for the environment and health. They cause disturbances in ecosystem functioning, are toxic for a wide range of non-target organisms, and have a high tendency to accumulate in the environment. Hence the development and discovery of new substances acting as pesticide are being derived from plants which are considered to be the best source of bioactive substance. Numerous herbs like Azadiractin indica, Allium sativum, Mentha piperita, Allium sativum, Artemisia absinthium, Citrullus colocynthis, Laurus nobilis, Mentha pulegium, Myrtus communis, Nerium oleander, Ocimum basilicum and Origanum majorana etc have shown pesticidal activity.

Melvin Blum et al (United States patent no. US5885600A) disclosed that the combination of neem, citronella and cedarwood oils made from cold processed extracted oils have insect repellent activity. They explained that pure cold processed crude neem oil has the same degree of repellent effectiveness as a 30% DEET formula.
A. Kumari et al [International Journal of Pharmaceutical Research, vol. 9(1), page no. 18] disclosed that different concentrations of lemongrass and neem seed oil containg formulations show synergistic action against the mosquitoes.
I.F. Mustafa et al [Nanomaterials, vol. 10(8), page no. 1608] disclosed the nanoemulsion preparation with herbal oils as active ingredient such as neem oil, citronella and lemongrass oil, thyme oil, sweet flag oil, eugenol oil, eucalyptus oil for pesticidal activity. This is formulation shows herbicidal,fungicidal and insecticidal activity.
K. Manhas et al (United States patent no. US20140242199A1) disclosed the various embodiments, based upon the natural oils such as neem oil, citronella oil, cinnamon oil, thyme oil, garlic oil lavender oil, geranium oil as pesticides.

From the above discussion we can say that a number of pesticide, synthetic as well as herbal, formulations are presently available with their various pros and cons. As for as synthetic drugs are concerned their cost-effectiveness, toxicity for humans are major issues. Most of the herbal pesticide formulations are presently not available in market extensively due to lack of the commercialization and absence of proper formulation which can be marketed easily. So there is still a need for a herbal formulation which has good herbicidal activity and which can be made available to the public in a suitable effective form.

OBJECT OF THE INVENTION

The main object of the present invention is to provide herbal pesticide formulations in dispersible tablet and microemulsion form which has very good pesticidal activity and which can be easily made available to the farmers for use. The proposed herbal formulations are also cheap in comparison to existing synthetic pesticides and fertilizers.
The another priniciple object of the present invention is to provide a herbal pesticide comprising of Neem (Azadiractica indica) seed oil, Lemongrass (Cybopogon citratus) oil and Bajra (Pennisetum typhoides) flour in dispersible tablet and microemulsion formulation.
Another principle object of present invention is to provide a process for preparation of such herbal pesticide formulations.

SUMMARY OF THE INVENTION

The present invention relates to the herbal pesticide formulations in dispersible tablet and microemulsion form which has very good pesticidal activity and which can be easily made available to the farmers for use.
The principle embodiment of the present invention provides a herbal pesticide formulation in microemulsion form comprising active ingredient as oil from seed of Azadiractica indica, oil from leaves of Cymbopogon citratus and flour of grains from Pennisetum typhoides. Excipients include tween 80S and oleic acid or citric acid, tartaric acid and sodium bicarbonate.
The another principle embodiment of the present invention provides a herbal pesticide formulation in microemulsion form comprising active ingredient as oil from seed of Azadiractica indica, oil from leaves of Cymbopogon citratus and flour of grains from Pennisetum typhoides. Excipients include tween 80S and oleic acid as emulsifying agents along with water.
The another principle embodiment of the present invention provides a herbal pesticide formulation in dispersible tablet form comprising active ingredient as oil from seed of Azadiractica indica, oil from leaves of Cymbopogon citratus and flour of grains from Pennisetum typhoides. Excipients include citric acid, tartaric acid and sodium bicarbonate as effervescent.
A further embodiment of present invention discloses a herbal pesticide formulation comprising
grain flour of Pennisetum typhoides 7.5 mg, Azadiracta indica seed oil 4 ml, Cymbopogon citratus leaves oil 4 ml, tween 80S 3 ml, oleic acid 3 ml and water 11 ml in a microemulsion form.
A further embodiment of present invention discloses a herbal pesticide formulation comprising Azadiractica indica seed oil 1 ml, Cybopogon citratus leaves oil 1 ml, grain flour of Pennisetum typhoides 1 gm, citric acid 1.8 gm, tartaric acid 2.7 gm and sodium bicarbonate 5.1 gm in a dispersible tablet form.
Another embodiment of present invention discloses the process for preparation of the herbal pesticide formulation in microemulsion form comprising steps of:
1. Mixing of seed oil of Azadiractica indica and leaves oil of Cybopogon citratus in 1:1 (4 ml each).
2. Adding of grain flour of Pennisetum typhoides (7.5 mg) in above mixture. Stirring well to form a homogenous mixture (mixture A).
3. Mixing 3 ml tween 80S and 3 ml oleic acid (1:1) in a separate beaker. Stirring well to form the mixture B.
4. Adding mixture B to mixture A on a sonicator.
5. Sonicating the resultant mixture for 15 minutes.
6. Adding 11 ml water and keeping the final mixture for 5 minutes followed by sonication to form a proper microemulsion.
Another embodiment of present invention discloses the process for preparation of the herbal pesticide formulation in dispersible tablet form comprising steps of:
1. Milling and mixing of 1.8 gm citric acid, 2.7 gm tartaric acid, 5.1 gm sodium bicarbonate and 1gm of grain flour of Pennisetum typhoides in mortar and pestle.
2. Preparing a wet mass of above powder mixture by using 1 ml seed oil of Azadiractica indica and 1 ml leaves oil of Cybopogon citratus as active ingredient and as solvent. Planetry mixer is used for wet mixing of the powders.
3. Forcing the wet mass of step 2 through a sieve (20 mesh screen) to produce wet granules.
4. Drying the wet granules in oven at 60 degree celcius temperature for 2 hrs.
5. Compressing the dried granules of step 4 by tablet press machine at a pressure of 6 tonnes.
BRIEF DESCRIPTION OF DRAWINGS
Figure 1 shows Crop image before application of formulation dose of 1.5 ml/Litre
Figure 2 shows Crop image after application of formulation at dose of 1.5 ml/Litre
Figure 3 shows Crop image before application of formulation dose of 2 ml/Litre
Figure 4 shows Crop image after application of formulation dose of 2 ml/Litre
Figure 5 shows Crop image before application of formulation dose of 2.5 ml/Litre
Figure 6 shows Crop image after application of formulation dose of 2.5 ml/Litre
Figure 7 shows Crop image before application of control
Figure 8 shows Crop image after application of control
DETAILED DESCRIPTION
In describing the embodiments of the invention, specific terminology is resorted for the terms so selected and it is to be understood that each specific term includes all technical equivalents that operate in a similar manner to accomplish a similar purpose.
Illustrative embodiments of the invention now will be described more fully hereinafter with reference to the examples, in which some, but not all embodiments of the invention are shown. Indeed, the invention may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will satisfy applicable legal requirements.
Pest insects can have adverse and damaging impacts on agricultural production and market access,the natural environment, and our lifestyle. Pest insects may cause problems by damaging crops and food production, parasitising livestock, or being a nuisance and health hazard to humans. Presently available synthetic pesticides have problem of the cost effectiveness and toxicity. Hence the development of new substances acting as pesticide are being derived through plants which are considered to be the best source of bioactive substances.
Azadirachta is a genus of two species of trees in the family Meliaceae. Numerous species have been proposed for the genus but only two are currently recognized, Azadirachta exceisa and the economically important tree Azadirachta indica, the Neem tree, from which neem oil is extracted. Both species are native to the Indomalaysian region, and A. indica is also widely cultivated and naturalized outside its native range. In traditional medicine in India, the resin from the trees have been attributed with medical benefits. A component in the resin which is an effective insecticide is azadiractin. Another component is an effective anti-fungal. Neem (Azadirachta indica) plant parts shows antimicrobial role through inhibitory effect on microbial growth/potentiality of cell wall breakdown.
Cymbopogon, also known as lemongrass, barbedgrass, silkyheads, cochingrass, Malabargrass, oilyheads or fevergrass, is a genus of Asian, African, Australian, and tropical island plants in the grassfamily. Some species (particularly Cymbopogon citratus) are commonly cultivated as culinary and medicinal herbs because of their scent, resembling that of lemons (Citrus limon). The name cymbopogon derives from the Greekwords kymbe and pogon which mean in most species, the hairy spikelets project from boat-shapeds pathes. Lemongrass is closely related to citronella, which is well known for being a great natural insect repellent. Lemongrass works on a wide variety of pests including chiggers, fleas, flies, fruitflies, no-see-ums, mosquitoes, and ticks.
Pennisetum is one of the most important genera of the family Poaceae. It includes such important species as pearlmillet, Pennisetumglaucum (L.) R. Brown [=Pennisetum typhoides (Burm.) StapfetHubb., Pennisetum americanum (L.), avaluable grain and forage crop; and its tetraploid relative Napiergrass (P.purpureum Schum.), prized for its fodder grown throughout the wet tropics of the world.

MATERIALS AND METHODS

Plant material
Grain flour of Pennisetum typhoides, Azadiracta indica seed oil, Cymbopogon citratus leaves oil and the excipients were collected from Meerut district.

Formulation of Dispersible Tablet
Ingredients (for a 11 gm batch size i.e. 11 tablets of 1 gm weight)
Azadiractica indica seed oil 1 ml (active constituent), Cybopogon citratus leaves oil 1 ml (active constituent), grain flour of Pennisetum typhoides 1 gm (active constituent), citric acid 1.8 gm (effervescent), tartaric acid 2.7 gm (effervescent) and sodium bicarbonate 5.1 gm (effervescent).

Process for the preparation of tablet
The process for preparation of the herbal pesticide formulation in dispersible tablet form comprising steps of:
1. Milling and mixing of 1.8 gm citric acid, 2.7 gm tartaric acid, 5.1 gm sodium bicarbonate and 1gm of grain flour of Pennisetum typhoides in mortar and pestle.
2. Preparing a wet mass of above powder mixture by using 1 ml seed oil of Azadiractica indica and 1 ml leaves oil of Cybopogon citratus as active ingredient and as solvent. Planetry mixer is used for wet mixing of the powders.
3. Forcing the wet mass of step 2 through a sieve (20 mesh screen) to produce wet granules.
4. Drying the wet granules in oven at 60 degree celcius temperature for 2 hrs.
5. Compressing the dried granules of step 4 by tablet press machine at a pressure of 6 tonnes.

Preformulation studies
The granules obtained for the trial batch were satisfactory. No rat holing was observed during the flow of granules from the hopper. Capping and sticking was not observed. From the compressibility index and Hausner’s ratio values obtained for granules of trial batch, the granules were found to have good flow properties.

Disintegration time of the formulated dispersible tablet
The formulated dispersible tablet was found to be disintegrated with 2 minutes in distilled water as well as in tap water at room temperature (25 degree celcius
Formulation of microemulsion
Ingredients (for a batch of 25 ml)
Grain flour of Pennisetum typhoides 7.5 mg (active constituent), Azadiracta indica seed oil 4 ml (active constituent), Cymbopogon citratus leaves oil 4 ml (active constituent), tween 80S 3 ml (emulsifying agent), oleic acid 3 ml (emulsifying agent) and water 11 ml.

Process for the preparation of microemulsion
The process for preparation of the herbal pesticide formulation in microemulsion form comprising steps of:
1. Mixing of seed oil of Azadiractica indica and leaves oil of Cybopogon citratus in 1:1 (4 ml each).
2. Adding of grain flour of Pennisetum typhoides (7.5 mg) in above mixture. Stirring well to form a homogenous mixture (mixture A).
3. Mixing 3 ml tween 80S and 3 ml oleic acid (1:1) in a separate beaker. Stirring well to form the mixture B.
4. Adding mixture B to mixture A on a sonicator.
5. Sonicating the resultant mixture for 15 minutes.
6. Adding 11 ml water and keeping the final mixture for 5 minutes followed by sonication to form a proper microemulsion.

Stability study of microemulsion
The physical stability of microemulsion was investigated via clarity and phase separation, which was observed at 2–8°C and at room temperature for 3 months. Microemulsion was found to be stable.

PESTICIDE TRIAL REPORT OF MICROEMULSION FORMULATION

CROP DETAILS
Name of crop: Brinjal Variety: Daftari- Manjusha
Date of Dosing: 20/01/2021 Irrigated / Non-Irrigated / Drip
Crop Stage: Vegetative Growth

EXPERIMENT DETAILS

Product Code/Name: Insecticide sample
Method of application: Foliar Spraying
Doses tested: 1 ml/Liter; 1.5 ml/Liter; 2.0 ml/Liter; 2.5 ml/Liter

Observation Data Sheet: Table:1

S.No Doses- 1 ml/Liter 1.5 ml/Liter 2.0 ml/Liter 2.5 ml/Liter Control

Observation- 0

Days 8

Days 0

Days 8

Days 0

Days 8

Days 0

Days 8

Days 0

Days 8

Days
1 Plant Height cm 60.24 73.26 58.26 68.16 59.58 69.2 58.12 78.0 57.82 63.78
2 No. of leaves 37.8 46.2 35.6 41.4 34.4 39.8 31.8 46.0 33 37.6
3 No. of new Shoots 19.2 23.6 18.0 20.8 18.8 22.0 17.4 25.0 17.2 19.2
4 No. of New Branches 4.0 5.8 4.0 6.0 3.8 6.5 4.0 10.0 3.4 4.4
5 No. of flower 0.0 4.0 0 6.0 1.0 8.0 0.0 12.0 0.0 2.0
6. No. of white flies’ population 15.0 12.0 10.0 9.0 10.0 8.0 10.0 6.0 10.0 12.0

Results
The results showed significant vegetative and reproductive growth in brinjal crop in terms of different parameters. There was also significant reduction in white flies population in the crops treated with 2.0 ml/Liter and 2.5 ml/Liter of the formulation sample.

Table 2: Insecticidal and growth promotor activity of microemulsion formulation
PESTICIDE TRIAL REPORT OF DISPERSIBLE TABLET FORMULATION

EXPERIMENT DETAILS

Product Name: Dispersible tablet
application: Spraying
Doses tested: 1 tablet weighing 750 mg dispersed in 100 ml of water
Insects: White aphids and Lady bug
Results: Insects were paralysed immediately after application and were dead within 10 minutes.

ANTIMICROBIAL ACTIVITY OF MICROEMULSION FORMULATION
Method used: Agar well diffusion method
Test Duration: Feb 22, 2021 to Feb 25, 2021.
Results: Result are described in Table 3.
Table 3: Antimicrobial activity of microemulsion formulation
SAMPLE CODE STRAIN
YB26 YB25 YB24 YB23 YB21
Control - - - - -
Microemulsion Formulation + + + + +
+ = Zone of inhibition was observed
- = No Zone of inhibition was observed
YB26 = Pseudomonas Sp.
YB25 = Staphylococcus Sp.
YB24 = Candida Sp.
YB23 = Pseudomonas Sp.
YB21 = Bacillus Subtilis

Claims:

WE CLAIM:
1. A herbal pesticide, herbal growth promoter and antimicrobial formulation comprising;
a) seed oil of Azadiracta indica;
b) leaves oil of Cymbopogon citratus;
c) grain flour of Pennisetum typhoides;
d) tween 80S;
e) oleic acid; and
f) water
wherein this formulation is in a microemulsion form.

2. A herbal pesticide, herbal growth promoter and antimicrobial formulation comprising;
a) seed oil of Azadiracta indica;
b) leaves oil of Cymbopogon citratus;
c) grain flour of Pennisetum typhoides;
d) citric acid;
e) tartaric acid; and
f) sodium bicarbonate
wherein this formulation is in a dispersible tablet form.

3. The herbal pesticide, herbal growth promoter and antimicrobial formulation as claimed in claim 1 wherein 25 ml of microemulsion formulation comprises 7.5 mg grain flour of Pennisetum typhoides, 4 ml seed oil of Azadiracta indica and 4 ml leaves oil of Cymbopogon citratus.

4. The herbal pesticide, herbal growth promoter and antimicrobial formulation as claimed in claim 1 wherein 25 ml of microemulsion comprises 3 ml of tween 80S and 3 ml of oleic acid as emulsifying agents, and 11 ml water.

5. The herbal pesticide, herbal growth promoter and antimicrobial formulation as claimed in claim 2 wherein a batch of 11 gm (11 tablets of 1 gm each) comprises 1 ml seed oil of Azadiractica indica, 1 ml leaves oil of Cybopogon citratus and 1 gm grain flour of Pennisetum typhoides.

6. The herbal pesticide, herbal growth promoter and antimicrobial formulation as claimed in claim 2 wherein a batch of 11 gm (11 tablets of 1 gm each) comprises 1.8 gm of citric acid, 2.7 gm of tartaric acid and 5.1 gm of sodium bicarbonate as effervescent.

7. The herbal pesticide, herbal growth promoter and antimicrobial formulation as claimed in claim 1 wherein microemulsion formulation is prepared by a process comprising steps of;
a. Mixing of seed oil of Azadiractica indica and leaves oil of Cybopogon citratus in 1:1 (4 ml each).
b. Adding of grain flour of Pennisetum typhoides (7.5 mg) in above mixture. Stirring well to form a homogenous mixture (mixture A).
c. Mixing 3 ml tween 80S and 3 ml oleic acid (1:1) in a separate beaker. Stirring well to form the mixture B.
d. Adding mixture B to mixture A on a sonicator.
e. Sonicating the resultant mixture for 15 minutes.
f. Adding 11 ml water and keeping the final mixture for 5 minutes followed by sonication to form a proper microemulsion.

8. The herbal pesticide, herbal growth promoter and antimicrobial formulation as claimed in claim 1 wherein dispersible tablet formulation is prepared by a process comprising steps of;
a. Milling and mixing of 1.8 gm citric acid, 2.7 gm tartaric acid, 5.1 gm sodium bicarbonate and 1 gm of grain flour of Pennisetum typhoides in mortar and pestle.
b. Preparing a wet mass of above powder mixture by using 1 ml seed oil of Azadiractica indica and 1 ml leaves oil of Cybopogon citratus as active ingredient and as solvent. Planetry mixer is used for wet mixing of the powders.
c. Forcing the wet mass of step b through a sieve (20 mesh screen) to produce wet granules.
d. Drying the wet granules in oven at 60 degree celcius temperature for 2 hrs.
e. Compressing the dried granules of step d by tablet press machine at a pressure of 6 tonnes.