Title of Invention	" A PROCESS FOR THE PREPARATION OF A SYNERGISTIC COMPOSITION USEFUL AS FUMIGANT AGAINST STORED GRAIN INSECT PESTS"
Abstract	A process for preparation of a synergistic composition useful as fumigant against stored grain insect-pests which comprises mixing by conventional manner such as herein described, 0.02 to 1.5%(wt) of 1, 8 cineole, obtained from the plant Eucalpytus globulus and 0.02 to 2.0%(wt) of one or more constituents of essential oil, obtained from plant species such as Ocimum basilicum, Mentaha spp., Cymbopogon spp., Anethum graveolens, Zingiber officnale, Pelargonium graveolens Cinnamomum spp., Eugenia spp., and Hyptis sp., wherein preferred ratio of said constituents ranges from 1:1:1 to 4:5:3.

Title of Invention

" A PROCESS FOR THE PREPARATION OF A SYNERGISTIC COMPOSITION USEFUL AS FUMIGANT AGAINST STORED GRAIN INSECT PESTS"

Abstract

A process for preparation of a synergistic composition useful as fumigant against stored grain insect-pests which comprises mixing by conventional manner such as herein described, 0.02 to 1.5%(wt) of 1, 8 cineole, obtained from the plant Eucalpytus globulus and 0.02 to 2.0%(wt) of one or more constituents of essential oil, obtained from plant species such as Ocimum basilicum, Mentaha spp., Cymbopogon spp., Anethum graveolens, Zingiber officnale, Pelargonium graveolens Cinnamomum spp., Eugenia spp., and Hyptis sp., wherein preferred ratio of said constituents ranges from 1:1:1 to 4:5:3.

Full Text	The present invention relates to a process for the preparation of a composition useful as fumigant against stored grain insect-pests. The formulation of the present invention has narcotic (fumigant) action against stored grain insects, Callosobruchus maculatus and Tribolium castaneum. The property is attributed to the synergistic combination of pure natural compound 1,8-cineol and other compounds isolated from essential oil bearing plants. Food commodities are considered to be first basic human need which are significantly damaged by insects during preharvest, post harvest and storage conditions, thereby making both qualitative and quantitative losses. Various stored commodities like rice, maize, wheat, legume, soyabean etc. contribute more than two third of the carbohydrate, half of protein, half of fiber and a quarter of the oil and fat requirements of everyday life (Evan, 1975, Crop Physiology, Cambridge). But these stored commodities when infested by insects, make a significant loss and degradation of food quality. Such infested grains with insects become totally unfit for human consumption. Hitherto known insecticides for control of stored grain insects are malathion, aluminium phosphide, methyl bromide and ethyl dibromide etc. They are totally synthetic and hazardous in one way or other. The use of these present day synthetic insecticide under storage conditions are highly unsafe due to increasing resistance, bioconcentration and residual problems. Introduction of synthetic pyrethroids clearly demonstrates the potential of this approach in terms of scientific and economic rewards (M. Elliot, ACS Series, 42, 1977). Apart from this, production of new chemicals has been declining in recent years and currently fewer than two insecticides are being introduction each year (Ware, Pesticides Theory and Application, San Fransico, 1983). Under the circumstances, replacement of "Conventional" insecticides with indigenous and environmentally friendly chemicals has the potential to provide adequate financial return and decreasing the environmental impact of synthetic chemicals apart from making substantial contribution towards increased global food production. During the course of our research, in an attempt for preparing insecticides from natural products,we have found that a natural essential oil constituent 1,8-cineol can be employed for controlling or killing the stored grain insect pulse beetle and other beetles. The technical ingredient used in the present invention, 1,8-cineole occurrs in more than 260 essential oils (Ganapathi, Current Science, 1937, 6:19). It is a colourless optically active liquid possessing a camphoraceous odour with a spicy cooling taste. It is used to combat flies, mosquitoes, bedbugs and chiggers etc. (Jacobson and Haller,1947,• Maugh.1982). It has antiseptic and expectorant activity and is used in many oily nose and throat sprays. 1,8-cineole can be prepared synthetically by the dehydration of terpene hydrate. It is also used widely in pharmaceutical preparations, applied internally or locally. Furthermore, it is used in room sprays, lotions and in all kinds of cosmetic preparations etc. The main objective of the present invention is to provide a formulation useful as fumigant against stored grain insect-pests which obviates the drawbacks as detai1ed above. Another object of the present invention is to provide a composition of the essential oil constituents as a formulation which is useful as protectant for the control of stored grain insects; which is safe, cheap, biodegradable, environmentally friendly, without any residual effect and commercially viable. Still another object of the present invention is to provide a composition of the volatile oil constituents as a formulation which is useful as stored grain protectant; which also take care of microbes, hot-spots produced in grains as a result of insect infestation during storage conditions due to its known fungal properties. The basis by which the invention has been developed is due to our finding that when 1,8-cineole isolated from essential oil bearing plants is mixed with one or more other natural essential oil constituents; the resultant composition develops insecticidal property which is especially active against stored grain insects. Accordingly, the composition is useful as fumigant for control of stored grain insects. Accordingly, the present invention provides a process for the preparation of a synergistic coi\mposition useful as fumigant against stored grain insect-pests which comprises mixing by conventional manner such as herein described 0.02 to 1.5% (wt) of 1, 8 cineole obtained from the plant Eucalyptus globulus and 0.02 to 2.0% (wt) of one or more constituents of essential oil obtained from plant species such as Ocimum basilicum, Mentha spp., Cymbopogon spp., Anethum graveolens, Zingiber officinale, Pelargonium graveolens, Cinnamomum spp., Eugenia spp., and Hyptis sp., wherein preferred ratio of said constituents ranges from 1:1:1 to 4:5:3. The volatile oil constituents used may be 1,8-cineole, linalool, menthol, carvone, methyl chavicol, anethole, citronellol, limonoids, curcumene and citral from plant genera Eucalyptus globulus, Ocimum basilicum, Mentha spp., Cymbopogon spp., Anethum graveolens, Coriandrum sativum, Foeniculum vulgare, Zingibe- officinale and Pelargonium graveolens. In the present invention a formulation useful as an fumigant against, stored grain insect-pests comprises blending the essential oil constituent 1.8-cineol extracted from the plant of the genus Eucalypts globulus with one or more essential natural oil constituent(s). In an embodiment of the present invention, has several components which include sustained release of aroma from the prepared composition of aromatic constituents to be effective as respiratory poision in killing certain stored grain insects like pulse beetle and red flour beetle attacking chickpea and wheat respectively; shooting down effect and rate of absorption in grains is very slow; shooting down effect ' in terms of exposure period under space trial test; safety in handling and minimised risk to human health. The present invention thereby provides for the preparation of a herbal fumigant which when used in stored grains kept in airtight storage conditions provide complete protection from insect-pest damage. The response is the result of aromatic properties of the product enabling respiratory arrest in target insects. The preferred composition of the present invention consists of pure volatile oil constituent 1,8-cineole , linalool and methyl chavicol isolated from plants. The amount of these components may range from 1:1:1 to 4:5:3. The invention is illustrated by the following examples which should not be construed to limit the sope of the present invention. EXAMPLE - 1 1,8-cineole of 98% purity obtained, from essential oil of Eucalyptus globulus was evaluated for the fumigant action at a dose of 0.3 ml./litre space against stored grain insects, pulse beetle (Ca11osobruchu3 maculatus) and red flour beetle (Tribo 1 iurn castaneum) . Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above liquid compound was applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication. Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds. It was observed that 1,8-eineole required 97,0 and 142=0 minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above. EXAMPLE - 2 Another pure essential oil constituent linalool isolated from plant genus Ocimum basilicum was evaluated for the fumigant action at a dose of 3 ml/litre space against stored grain insects, pulse beetle (Ca11osobruchus maculatus) and red flour beetle (Tribol ium castaneurn) . Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above liquid compound was applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication. Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds. It was observed that linalol required 155.0 and 198.0 minutes to Xill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above. EXAMPLE - 3 Another pure essential oil constituent methyl chavicol isolated from plant genus Ocimum basil 1icum was evaluated for the fumigant action at a dose of 3 ml/litre space against stored grain insects, pulse beetle (Callosobruchus maculatus) and red flour beetle (Tribolium castaneum). Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above liquid compound was applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication. Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds. It was observed that methyl chavicol required 212=0 and 310.0 minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above. EXAMPLE - 4 1,8-cineole (8.0 ml.) of 98% purity obtained from essential oil of Eucalyptus globulus is mixed with another volatile oil constituent linalool (8.0 ml.) in ratio of 1:1 to make a composition and evaluated for the fumigant action at a dose of 3 ml/litre space against stored grain insects, pulse beetle (Ca11osobruchus maculatus) and red flour beetle (Tribolium castaneum). Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above liquid compound was applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication. Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds. It was observed that above composition required 135.0 and 180.0 minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above. EXAMPLE - 5 1.8-cineole (8.0 ml.) of 98% purity obtained from essential oil of Eucalyptus globulus is mixed with another volatile oil constituent methyl chavicol and evaluated for the fumagant action at a dose of 3 ml/litre space against stored grain insects, pulse beetle (Callosobruchu3 maculatus) and red flour beetle (Tribo1ium ca3taneum). Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above liquid compound was applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication. Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds. It was observed that the above composition required 176.0 and 22D.U minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above. EXAMPLE-6 1,8-cineole (5.0 ml.) of 98* purity obtained from essential oil of Eucalyptus q1obu1us is mixed with compound linalool (5.0 ml.) and compound methyl chavicol (5.0 ml.) to make a composition in the ratio of 1:1:1 and evaluated for the fumigant action at a dose of 3 ml/litre space against stored grain insects, pulse beetle (Ca11osobruchus maculatus) and red flour beetle (Tribolium castaneum). Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above composition is applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication. Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds. It was observed that the above composition required 85.0 and 135.0 minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above. EXAMPLE-7 1,8-cineole (5.0 ml.) of 98* purity obtained from essential oil of Eucalyptus globulus is mixed with compound linalool (10.0 ml.) and compound methyl chavicol (10.0 ml.) to make a composition in the ratio of 1:2:2 and evaluated for the fumigant action at a dose of 3 ml/litre space against stored grain insects, pulse beetle (Callosobruchus maculatus) and red flour beetle (Tribolium castaneum). Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above composition is applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication. Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds. It was observed that the above composition required 107.0 and 176.0 minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above. EXAMPLE-8 1,8-cineole (10.0 ml.) of 98% purity obtained from essential oil of Eucalyptus globulus is mixed with compound linalool (15.0 ml.) and compound methyl chavicol (5.0 ml.) to make a composition in the ratio of 2:3:1 and evaluated for the fumigant action at a dose of 3 ml/litre space against stored grain insects, pulse beetle (Callosobruchus maculatus) and red flour beetle (Tribolium castaneum). Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above composition is applied on it. The treated blotting paper was Kept in empty pearl pet jar of one litre capacity and considered as one replication. Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds. It was observed that the above composition required 80.0 and 105.0 minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above. EXAMPLE-9 1,8-cineole (15.0 ml.) of 98* purity obtained from essential oil of Eucalyptus globulus is mixed with compound linalool (10.0 ml.) and compound methyl chavicol (5.0 ml.) to make a composition in the ratio of 3:2:1 and evaluated for the fumigant action at a dose of 3 ml/litre space against stored grain insects, pulse beetle (Ca11osobruchus maculatus) and red flour beetle (Tribo1iurn castaneum). Bioassay was done by using ordinary blotting paper of sise 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above composition is applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication. Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds. It was observed that the above composition required 68.0 and 120.0 minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above. EXAMPLE-10 1.8-cineole (20.0 ml.) of 98% purity obtained from essential oil of Eucalyptus globulus is mixed with compound linalool (25.0 ml.) and compound methyl chavicol (15.0 ml.) to make a composition in the ratio of 4:5:3 and evaluated for the fumigant action at a dose of 3 ml/litre space against stored grain insects, pulse beetle (Ca11osobruchus maculatus) and red flour beetle (Tribolium castaneum). Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above composition is applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication. Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds. It was observed that the above composition required 94.0 and 112.0 minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above. EXAMPLE - 11 This examples relates to the effect of a known insecticide so as to compare the results with that of the composition of the present invention. Aluminium phosphide is a synthetic fumigant commercially available in the form of tablet or powder in the market under the trade name Fumino bags or Celphos tablets. One tablet was kept at the bottom of an empty pearl pet jar (1.0 lit.) and considered as one replication. Fifty insects of pulse beetle were released and mouth of the jar was covered by its lid airtightly. The time required to kill 99.0 per cent pulse beetles in the jar was recorded and considered as exposure period to kill 99.00 per cent insects at a fixed dosage and volume. Five replications were used for the observation of shooting down effect for a total of 250 insects. Same procedure was followed for red flour beetle using wheat seeds. It was observed that one tablet required 65 minutes to kill 99.0 per cent of both the insects at a fixed dosage and volume as stated above. The summary of results of above examples (1-11) are given in Table - 1. Table 1: Effect of pure essential oil constituents or their mixtures as fumigant against two stored grain insects, pulse beetle and red flour beetle. (Table Removed) It is observed from the above table that the compound 1,8-cineol when mixed with other compounds linalool and methyl chavicol into stored grain protectant formulation at varying degree of ratio, resulted into varying degree of shooting down effect. Accordingly it can be observed that the composition of the present invention is not a mere admixture having the aggregate properties of all the ingredients but is a synergistic mixture resulting in enhanced property with sustained effect. EXAMPLE - 12 In order to observe phytotoxic effects of the composition prepared. in terms of seed germination, viability test was carried out: Chickpea aeede (50 am) were taken in a beaker and 1 ml. of the composition having ingredients 1,8-cineole (2.0 ml.), linalool (2.0 ml) and methyl chavicol (2.0 ml.) (ratio of 1:1:1) was mixed and stirred for 10 minutes and kept under room conditions to get dried. Thus treated seeds were sown in earthen pots. Five replications were used with 50 gm seeds per replication. Germination of the seeds was recorded for 7 days. Same procedure was followed for wheat seeds. The experimental data revealed 100.00 per cent germination of chickpea and wheat seeds when treated with above composition. EXAMPLE - 13 In order to observe phytotoxic effects of the composition prepared, in terms of seed germination, viability test was carried out. Chickpea seeds (50 gm) were taken in a beaker and 1 ml. of the composition having ingredients 1,8-cineole (2.0 ml.), linalool (4.0 ml) and methyl chavicol (4.0 ml.) (ratio of 1:2:2) was mixed and stirred for 10 minutes and kept under room conditions to get dried. Thus treated seeds were sown in earthen pots. Five replications were used with 50 gm seeds per replication. Germination of the seeds was recorded for 7 days. Same procedure was followed for wheat seeds. 20 The experimental data revealed 100.00 per cent germination of chickpea and wheat seeds when treated with above composition. EXAMPLE - 14 In order to observe phytotoxic effects of the composition prepared, in terms of seed germination, viability test was carried out. Chickpea seeds (50 gm) were taken in a beaker and 1 ml. of the composition having ingredients 1,8-cineole (4.0 ml.), linalool (6.0 ml) and methyl chavicol (2.0 ml.) (ratio of 2:3:1) was mixed and stirred for 10 minutes and kept under room conditions to get dried. Thus treated seeds were sown in earthen pots. Five replications were used with 50 gm seeds per replication. Germination of the seeds was recorded for 7 days. Same procedure was followed for wheat seeds. The experimental data revealed 100.00 per cent germination of chickpea and wheat seeds when treated with the above compos i t i on. EXAMPLE - 15 In order to observe phytotoxic effects of the composition prepared, in terms of seed germination, viability test was carried out. Chickpea seeds (50 gm) were taken in a beaker and 1 ml. of the composition having ingredients 1,8-cineole (6.0 ml.), linalool (4.0 ml) and methyl chavicol (2.0 ml.) (ratio of 3,2-1) was mixed and stirred for 10 minutes and kept under room conditions to get dried. Thus treated seeds were sown in earthen pots. Five replications were used with 50 gm seeds per replication. Germination of the seeds was recorded for 7 days. Same procedure was followed for wheat seeds. The experimental data revealed' 100.00 per cent germination of chickpea and wheat seeds when treated with the above composition. EXAMPLE - 16 In order to observe phytotoxic effects of the composition prepared, in terms of seed germination, viability test was carried out. Chickpea seeds (50 gm) were taken in a beaker and 1 ml. of the composition having ingredients 1,8-cineol (8.0 ml.), linalool (10.0 ml) and methyl chavicol (6.0 ml.) (ratio of 4:5:3) was mixed and stirred for 10 minutes and kept under room conditions to get dried. Thus treated seeds were sown in earthen pots. Five replications were used with 50 gm seeds per replication. Germination of the seeds was recorded for 7 days. Same procedure was followed for wheat seeds. The experimental data revealed 100.00 per cent germination of chickpea and wheat seeds when treated with the above composition. EXAMPLE - 17 This example relates to the phytotoxic effect of a known insecticide so as to compare the results with that of the composition of the present invention. Aluminium phosphide is a synthetic fumigant commercially available in the form of tablets. One tablet of Celphos was kept in a one litre capacity pearl pet jar. Fifty gram of chickpea seed was placed in it and mouth of the jar was covered with its lid. After one hour, seeds were taken out and kept at room temperature for 24 hours. After that treated gram seeds were sown in earthen pots. Five replication were used with 50 gm. seed per replication. Same procedure was followed in case of wheat seeds. Germination of the seed was recorded after 7 days. Experimental data revealed 100.00 percent germination of the treated seeds. Summary of the above example (12-17) is given in table - 2. Tattle 8 Effect of pure essential oil constituent in combination product on the germination of chickpea and wheat seeds. (Table Removed) Table-2 clearly indicates that preparation of this invention has no phytotoxic effect in terms of germination of chickpea and wheat seeds. The advantage of present invention is that being plant based formulation, it is eco-friendly, non-toxic, non-corrosive and non-phytotoxic apart from sustained and shooting down effect and safe in use. We claim: 1. A process for the preparation of a synergistic composition useful as fumigant against stored grain insect-pests which comprises mixing by conventional manner such as herein described, 0.02 to 1.5% (wt) of 1, 8 cineole, obtained from the plant Eucalyptus globulus and 0.02 to 2.0% (wt) of one or more constituents of essential oil; obtained from plant species such as Ocimum basilicum, Mentha spp., Cymbopogon spp., Anethum graveolens, Zingiber officinale, Pelargonium graveolens, Cinnamomum spp., Eugenia spp., and Hyptis sp., wherein preferred ratio of said constituents ranges from 1:1:1 to 4:5:3. 2. A process as claimed in claim 1 wherein the constituents of essential oil used is selected from linalool, methyl chavicol, menthol, carvone, anethole, citronellol, citral, and curcumene 3. A process for the preparation of a synergistic composition useful as fumigant against stored grain insect-pests substantially as herein described with reference to the examples nos 6-10

Full Text

The present invention relates to a process for the preparation of a composition useful as fumigant against stored grain insect-pests.
The formulation of the present invention has narcotic (fumigant) action against stored grain insects, Callosobruchus maculatus and Tribolium castaneum. The property is attributed to the synergistic combination of pure natural compound 1,8-cineol and other compounds isolated from essential oil bearing plants.
Food commodities are considered to be first basic human need which are significantly damaged by insects during preharvest, post harvest and storage conditions, thereby making both qualitative and quantitative losses. Various stored commodities like rice, maize, wheat, legume, soyabean etc. contribute more than two third of the carbohydrate, half of protein, half of fiber and a quarter of the oil and fat requirements of everyday life (Evan, 1975, Crop Physiology, Cambridge). But these stored commodities when infested by insects, make a significant loss and degradation of food quality. Such infested grains with insects become totally unfit for human consumption.
Hitherto known insecticides for control of stored grain insects are malathion, aluminium phosphide, methyl bromide and ethyl dibromide etc. They are totally synthetic and hazardous in one way or other. The use of these present day

synthetic insecticide under storage conditions are highly unsafe due to increasing resistance, bioconcentration and residual problems.
Introduction of synthetic pyrethroids clearly demonstrates the potential of this approach in terms of scientific and economic rewards (M. Elliot, ACS Series, 42, 1977). Apart from this, production of new chemicals has been declining in recent years and currently fewer than two insecticides are being introduction each year (Ware, Pesticides Theory and Application, San Fransico, 1983).
Under the circumstances, replacement of "Conventional" insecticides with indigenous and environmentally friendly chemicals has the potential to provide adequate financial return and decreasing the environmental impact of synthetic chemicals apart from making substantial contribution towards increased global food production.
During the course of our research, in an attempt for preparing insecticides from natural products,we have found that a natural essential oil constituent 1,8-cineol can be employed for controlling or killing the stored grain insect pulse beetle and other beetles.
The technical ingredient used in the present invention, 1,8-cineole occurrs in more than 260 essential oils (Ganapathi, Current Science, 1937, 6:19). It is a colourless optically

active liquid possessing a camphoraceous odour with a spicy cooling taste. It is used to combat flies, mosquitoes, bedbugs and chiggers etc. (Jacobson and Haller,1947,• Maugh.1982). It has antiseptic and expectorant activity and is used in many oily nose and throat sprays. 1,8-cineole can be prepared synthetically by the dehydration of terpene hydrate.
It is also used widely in pharmaceutical preparations, applied internally or locally. Furthermore, it is used in room sprays, lotions and in all kinds of cosmetic preparations etc.
The main objective of the present invention is to provide a formulation useful as fumigant against stored grain insect-pests which obviates the drawbacks as detai1ed above.
Another object of the present invention is to provide a composition of the essential oil constituents as a formulation which is useful as protectant for the control of stored grain insects; which is safe, cheap, biodegradable, environmentally friendly, without any residual effect and commercially viable.
Still another object of the present invention is to provide a composition of the volatile oil constituents as a formulation which is useful as stored grain protectant;

which also take care of microbes, hot-spots produced in grains as a result of insect infestation during storage conditions due to its known fungal properties.
The basis by which the invention has been developed is due to our finding that when 1,8-cineole isolated from essential oil bearing plants is mixed with one or more other natural essential oil constituents; the resultant composition develops insecticidal property which is especially active against stored grain insects. Accordingly, the composition is useful as fumigant for control of stored grain insects.
Accordingly, the present invention provides a process for the preparation of a synergistic coi\mposition useful as fumigant against stored grain insect-pests which comprises mixing by conventional manner such as herein described 0.02 to 1.5% (wt) of 1, 8 cineole obtained from the plant Eucalyptus globulus and 0.02 to 2.0% (wt) of one or more constituents of essential oil obtained from plant species such as Ocimum basilicum, Mentha spp., Cymbopogon spp., Anethum graveolens, Zingiber officinale, Pelargonium graveolens, Cinnamomum spp., Eugenia spp., and Hyptis sp., wherein preferred ratio of said constituents ranges from 1:1:1 to 4:5:3.
The volatile oil constituents used may be 1,8-cineole, linalool, menthol,
carvone, methyl chavicol, anethole, citronellol, limonoids, curcumene and
citral from plant genera Eucalyptus globulus, Ocimum basilicum, Mentha
spp., Cymbopogon spp., Anethum graveolens, Coriandrum sativum,
Foeniculum vulgare, Zingibe- officinale and Pelargonium graveolens.

In the present invention a formulation useful as an fumigant against, stored grain insect-pests comprises blending the essential oil constituent 1.8-cineol extracted from the plant of the genus Eucalypts globulus with one or more essential natural oil constituent(s).
In an embodiment of the present invention, has several components which include sustained release of aroma from the prepared composition of aromatic constituents to be effective as respiratory poision in killing certain stored grain insects like pulse beetle and red flour beetle attacking chickpea and wheat respectively; shooting down effect and rate of absorption in grains is very slow; shooting down effect ' in terms of exposure period under space trial test; safety in handling and minimised risk to human health. The present invention thereby provides for the preparation of a herbal fumigant which when used in stored grains kept in airtight storage conditions provide complete protection from insect-pest damage. The response is the result of aromatic properties of the product enabling respiratory arrest in target insects.
The preferred composition of the present invention consists of pure volatile oil constituent 1,8-cineole , linalool and methyl chavicol isolated from plants. The amount of these components may range from 1:1:1 to 4:5:3.

The invention is illustrated by the following examples which should not be construed to limit the sope of the present invention.
EXAMPLE - 1
1,8-cineole of 98% purity obtained, from essential oil of Eucalyptus globulus was evaluated for the fumigant action at a dose of 0.3 ml./litre space against stored grain insects, pulse beetle (Ca11osobruchu3 maculatus) and red flour beetle (Tribo 1 iurn castaneum) .
Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above liquid compound was applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication.
Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds.

It was observed that 1,8-eineole required 97,0 and 142=0 minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above.
EXAMPLE - 2
Another pure essential oil constituent linalool isolated from plant genus Ocimum basilicum was evaluated for the fumigant action at a dose of 3 ml/litre space against stored grain insects, pulse beetle (Ca11osobruchus maculatus) and red flour beetle (Tribol ium castaneurn) .
Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above liquid compound was applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication.
Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds.

It was observed that linalol required 155.0 and 198.0 minutes to Xill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above.
EXAMPLE - 3
Another pure essential oil constituent methyl chavicol isolated from plant genus Ocimum basil 1icum was evaluated for the fumigant action at a dose of 3 ml/litre space against stored grain insects, pulse beetle (Callosobruchus maculatus) and red flour beetle (Tribolium castaneum).
Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above liquid compound was applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication.
Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds.

It was observed that methyl chavicol required 212=0 and 310.0 minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above.
EXAMPLE - 4
1,8-cineole (8.0 ml.) of 98% purity obtained from essential oil of Eucalyptus globulus is mixed with another volatile oil constituent linalool (8.0 ml.) in ratio of 1:1 to make a composition and evaluated for the fumigant action at a dose of 3 ml/litre space against stored grain insects, pulse beetle (Ca11osobruchus maculatus) and red flour beetle (Tribolium castaneum).
Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above liquid compound was applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication.
Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was

followed for another stored grain insect, red flour beetle using wheat seeds.
It was observed that above composition required 135.0 and 180.0 minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above.
EXAMPLE - 5
1.8-cineole (8.0 ml.) of 98% purity obtained from essential oil of Eucalyptus globulus is mixed with another volatile oil constituent methyl chavicol and evaluated for the fumagant action at a dose of 3 ml/litre space against stored grain insects, pulse beetle (Callosobruchu3 maculatus) and red flour beetle (Tribo1ium ca3taneum).
Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above liquid compound was applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication.
Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume.

Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds.
It was observed that the above composition required 176.0 and 22D.U minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above.
EXAMPLE-6
1,8-cineole (5.0 ml.) of 98* purity obtained from essential oil of Eucalyptus q1obu1us is mixed with compound linalool (5.0 ml.) and compound methyl chavicol (5.0 ml.) to make a composition in the ratio of 1:1:1 and evaluated for the fumigant action at a dose of 3 ml/litre space against stored grain insects, pulse beetle (Ca11osobruchus maculatus) and red flour beetle (Tribolium castaneum).
Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above composition is applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication.
Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The

time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds.
It was observed that the above composition required 85.0 and 135.0 minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above.
EXAMPLE-7 1,8-cineole (5.0 ml.) of 98* purity obtained from essential oil of Eucalyptus globulus is mixed with compound linalool (10.0 ml.) and compound methyl chavicol (10.0 ml.) to make a composition in the ratio of 1:2:2 and evaluated for the fumigant action at a dose of 3 ml/litre space against stored grain insects, pulse beetle (Callosobruchus maculatus) and red flour beetle (Tribolium castaneum).
Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above composition is applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication.

Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds.
It was observed that the above composition required 107.0 and 176.0 minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above.
EXAMPLE-8 1,8-cineole (10.0 ml.) of 98% purity obtained from essential oil of Eucalyptus globulus is mixed with compound linalool (15.0 ml.) and compound methyl chavicol (5.0 ml.) to make a composition in the ratio of 2:3:1 and evaluated for the fumigant action at a dose of 3 ml/litre space against stored grain insects, pulse beetle (Callosobruchus maculatus) and red flour beetle (Tribolium castaneum).
Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above composition is applied on it. The treated

blotting paper was Kept in empty pearl pet jar of one litre capacity and considered as one replication.
Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds.
It was observed that the above composition required 80.0 and 105.0 minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above.
EXAMPLE-9 1,8-cineole (15.0 ml.) of 98* purity obtained from essential oil of Eucalyptus globulus is mixed with compound linalool (10.0 ml.) and compound methyl chavicol (5.0 ml.) to make a composition in the ratio of 3:2:1 and evaluated for the fumigant action at a dose of 3 ml/litre space against stored grain insects, pulse beetle (Ca11osobruchus maculatus) and red flour beetle (Tribo1iurn castaneum).

Bioassay was done by using ordinary blotting paper of sise 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above composition is applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication.
Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds.
It was observed that the above composition required 68.0 and 120.0 minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above.
EXAMPLE-10 1.8-cineole (20.0 ml.) of 98% purity obtained from essential oil of Eucalyptus globulus is mixed with compound linalool (25.0 ml.) and compound methyl chavicol (15.0 ml.) to make a
composition in the ratio of 4:5:3 and evaluated for the fumigant action at a dose of 3 ml/litre space against stored

grain insects, pulse beetle (Ca11osobruchus maculatus) and red flour beetle (Tribolium castaneum).
Bioassay was done by using ordinary blotting paper of size 6x8 cm. in triplicate. A fixed amount of 3.0 ml. of the above composition is applied on it. The treated blotting paper was kept in empty pearl pet jar of one litre capacity and considered as one replication.
Fifty insects of pulse beetle were released inside the jar and mouth of the jar was covered airtightly by its lid. The time required to kill 99.0 per cent insect population in the jar was recorded and considered as exposure period to kill 99.0 per cent pulse beetles at a fixed dosage and volume. Five replications were used for observation of shooting down effect for the total of 250 insects. Same procedure was followed for another stored grain insect, red flour beetle using wheat seeds.
It was observed that the above composition required 94.0 and 112.0 minutes to kill 99.0 per cent of pulse beetle and red flour beetle respectively at a fixed dosage and volume as stated above.
EXAMPLE - 11
This examples relates to the effect of a known insecticide so as to compare the results with that of the composition of

the present invention. Aluminium phosphide is a synthetic fumigant commercially available in the form of tablet or powder in the market under the trade name Fumino bags or Celphos tablets. One tablet was kept at the bottom of an empty pearl pet jar (1.0 lit.) and considered as one replication.
Fifty insects of pulse beetle were released and mouth of the jar was covered by its lid airtightly. The time required to kill 99.0 per cent pulse beetles in the jar was recorded and considered as exposure period to kill 99.00 per cent insects at a fixed dosage and volume. Five replications were used for the observation of shooting down effect for a total of 250 insects. Same procedure was followed for red flour beetle using wheat seeds.
It was observed that one tablet required 65 minutes to kill 99.0 per cent of both the insects at a fixed dosage and volume as stated above.
The summary of results of above examples (1-11) are given in Table - 1.

Table 1: Effect of pure essential oil constituents or their mixtures as fumigant against two stored grain insects, pulse beetle and red flour beetle.
(Table Removed)

It is observed from the above table that the compound 1,8-cineol when mixed with other compounds linalool and methyl chavicol into stored grain protectant formulation at varying degree of ratio, resulted into varying degree of shooting down effect.
Accordingly it can be observed that the composition of the present invention is not a mere admixture having the aggregate properties of all the ingredients but is a synergistic mixture resulting in enhanced property with sustained effect.
EXAMPLE - 12
In order to observe phytotoxic effects of the composition prepared. in terms of seed germination, viability test was

carried out: Chickpea aeede (50 am) were taken in a beaker and 1 ml. of the composition having ingredients 1,8-cineole (2.0 ml.), linalool (2.0 ml) and methyl chavicol (2.0 ml.) (ratio of 1:1:1) was mixed and stirred for 10 minutes and kept under room conditions to get dried. Thus treated seeds were sown in earthen pots. Five replications were used with 50 gm seeds per replication. Germination of the seeds was recorded for 7 days. Same procedure was followed for wheat seeds.
The experimental data revealed 100.00 per cent germination of chickpea and wheat seeds when treated with above composition.
EXAMPLE - 13
In order to observe phytotoxic effects of the composition prepared, in terms of seed germination, viability test was carried out. Chickpea seeds (50 gm) were taken in a beaker and 1 ml. of the composition having ingredients 1,8-cineole (2.0 ml.), linalool (4.0 ml) and methyl chavicol (4.0 ml.) (ratio of 1:2:2) was mixed and stirred for 10 minutes and kept under room conditions to get dried. Thus treated seeds were sown in earthen pots. Five replications were used with 50 gm seeds per replication. Germination of the seeds was recorded for 7 days. Same procedure was followed for wheat seeds.
20

The experimental data revealed 100.00 per cent germination of chickpea and wheat seeds when treated with above composition.
EXAMPLE - 14
In order to observe phytotoxic effects of the composition prepared, in terms of seed germination, viability test was carried out. Chickpea seeds (50 gm) were taken in a beaker and 1 ml. of the composition having ingredients 1,8-cineole (4.0 ml.), linalool (6.0 ml) and methyl chavicol (2.0 ml.) (ratio of 2:3:1) was mixed and stirred for 10 minutes and kept under room conditions to get dried. Thus treated seeds were sown in earthen pots. Five replications were used with 50 gm seeds per replication. Germination of the seeds was recorded for 7 days. Same procedure was followed for wheat seeds.
The experimental data revealed 100.00 per cent germination of chickpea and wheat seeds when treated with the above compos i t i on.
EXAMPLE - 15 In order to observe phytotoxic effects of the composition prepared, in terms of seed germination, viability test was carried out. Chickpea seeds (50 gm) were taken in a beaker and 1 ml. of the composition having ingredients 1,8-cineole (6.0 ml.), linalool (4.0 ml) and methyl chavicol (2.0 ml.)

(ratio of 3,2-1) was mixed and stirred for 10 minutes and kept under room conditions to get dried. Thus treated seeds were sown in earthen pots. Five replications were used with 50 gm seeds per replication. Germination of the seeds was recorded for 7 days. Same procedure was followed for wheat seeds.
The experimental data revealed' 100.00 per cent germination of chickpea and wheat seeds when treated with the above composition.
EXAMPLE - 16 In order to observe phytotoxic effects of the composition prepared, in terms of seed germination, viability test was carried out. Chickpea seeds (50 gm) were taken in a beaker and 1 ml. of the composition having ingredients 1,8-cineol (8.0 ml.), linalool (10.0 ml) and methyl chavicol (6.0 ml.) (ratio of 4:5:3) was mixed and stirred for 10 minutes and kept under room conditions to get dried. Thus treated seeds were sown in earthen pots. Five replications were used with 50 gm seeds per replication. Germination of the seeds was recorded for 7 days. Same procedure was followed for wheat seeds.
The experimental data revealed 100.00 per cent germination of chickpea and wheat seeds when treated with the above composition.

EXAMPLE - 17
This example relates to the phytotoxic effect of a known insecticide so as to compare the results with that of the composition of the present invention. Aluminium phosphide is a synthetic fumigant commercially available in the form of tablets. One tablet of Celphos was kept in a one litre capacity pearl pet jar. Fifty gram of chickpea seed was placed in it and mouth of the jar was covered with its lid. After one hour, seeds were taken out and kept at room temperature for 24 hours. After that treated gram seeds were sown in earthen pots. Five replication were used with 50 gm. seed per replication. Same procedure was followed in case of wheat seeds.
Germination of the seed was recorded after 7 days. Experimental data revealed 100.00 percent germination of the treated seeds. Summary of the above example (12-17) is given in table - 2.

Tattle 8 Effect of pure essential oil constituent in
combination product on the germination of chickpea and wheat seeds.
(Table Removed)
Table-2 clearly indicates that preparation of this invention has no phytotoxic effect in terms of germination of chickpea and wheat seeds.
The advantage of present invention is that being plant based formulation, it is eco-friendly, non-toxic, non-corrosive and non-phytotoxic apart from sustained and shooting down effect and safe in use.

We claim:
1. A process for the preparation of a synergistic composition useful as
fumigant against stored grain insect-pests which comprises mixing by
conventional manner such as herein described, 0.02 to 1.5% (wt) of 1, 8 cineole,
obtained from the plant Eucalyptus globulus and 0.02 to 2.0% (wt) of one or
more constituents of essential oil; obtained from plant species such as Ocimum
basilicum, Mentha spp., Cymbopogon spp., Anethum graveolens, Zingiber
officinale, Pelargonium graveolens, Cinnamomum spp., Eugenia spp., and Hyptis
sp., wherein preferred ratio of said constituents ranges from 1:1:1 to 4:5:3.
2. A process as claimed in claim 1 wherein the constituents of essential oil used
is selected from linalool, methyl chavicol, menthol, carvone, anethole, citronellol,
citral, and curcumene
3. A process for the preparation of a synergistic composition useful as
fumigant against stored grain insect-pests substantially as herein described with
reference to the examples nos 6-10

Documents:

372-del-1998-abstract.pdf

372-del-1998-claims.pdf

372-del-1998-complete specification (granted).pdf

372-del-1998-correspondence-others.pdf

372-del-1998-correspondence-po.pdf

372-del-1998-description (complete).pdf

372-del-1998-form-1.pdf

372-del-1998-form-2.pdf

372-del-1998-form-4.pdf

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Patent Number

188378

Indian Patent Application Number

372/DEL/1998

PG Journal Number

37/2002

Publication Date

14-Sep-2002

Grant Date

20-Jun-2003

Date of Filing

13-Feb-1998

Name of Patentee

COUNCIL OF SCIENTIFIC AND INDUSTRIAL RESEARCH

Applicant Address

RAFI MARG, NEW DELHI-110001 INDIA.

Inventors:

#	Inventor's Name	Inventor's Address
1	SUSHIL KUMAR	CENTRAL INSTITUTE OF MEDICINAL & AROMATIC PLANTS LUCKNOW-226015, INDIA
2	VEENA PRAJAPATI	CENTRAL INSTITUTE OF MEDICINAL & AROMATIC PLANTS LUCKNOW-226015, INDIA
3	KISHAN KUMAR AGGARWAL	CENTRAL INSTITUTE OF MEDICINAL & AROMATIC PLANTS LUCKNOW-226015, INDIA
4	SUMAN PREET SINGH KHANUJA	CENTRAL INSTITUTE OF MEDICINAL & AROMATIC PLANTS LUCKNOW-226015, INDIA
5	ARUN KUMAR TRIPATHI	CENTRAL INSTITUTE OF MEDICINAL & AROMATIC PLANTS LUCKNOW-226015, INDIA.

PCT International Classification Number

A01N 65/00

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA