DESC:FORM – 2

THE PATENTS ACT, 1970
(39 OF 1970)

COMPLETE SPECIFICATION
(SECTION 10, RULE 13)

“A PROCESS FOR PREPARATION OF NANO STRUCTURED NATURAL AND HYBRID COMPOSITE PLANT NUTRIENTS”

RAY NANO SCIENCE AND RESERCH CENTRE LLP
Having address at
Plot no. 7 & 8, GIDC, V U Nagar,
Anand, Gujarat, India - 388001

The following specification particularly describes the nature of this invention and the manner in which it is performed:

FIELD OF INVENTION:
The present invention relates to a process for the preparation of nano structured natural and hybrid composites of plant nutrient formulation. Moreover, present invention relates to a process for the preparation of nano structured natural and hybrid composites of plant nutrient formulation using cow urine and/or its distillate as dispersing medium.
BACKGROUND OF INVENTION:
Increased food demands and ceasing nutrient deposits have resulted in a great loss between the food supply and demands over the period of time and this same phenomenon continues further. Due to higher inputs of synthetic fertilizers on lands, greater impacts have been resulted and observed in environment leading to physicochemical conditions of environmental pollution as well as persistent changes in the soil ecology. This impact has greatly decreased the fertility of natural soil thereby hindering the agricultural productivity, human health, and hygiene.
Chemical fertilizers are unsustainable for a variety of reasons, including high application inefficiency in terms of plant tissue drying up, massive energy and input costs for manufacture, and possibly severe environmental consequences. Food security attainment and sustainability are critical global concerns that necessitate adapted agricultural practices to effectively counteract the implications of climate change, population growth, and arable land loss. Unlike commercial chemical fertilizers, cow urine or ark fortified with natural and/or inorganic nutrients-based fertilizers have the potential to improve crop output growth and production without overfertilization.
The most preferred choice of fertilizer by farmers is synthetic fertilizer. This choice of selection is due to various reasons, which includes ease of application, availability, proven dosage usage, etc. However, various problems like less nutrient efficiency, leaching, volatilization, have been a continuous threat to the ecosystem, which has exponentially increased over the years due to population growth and to overcome the demand-supply gap.
Cow urine and its distillates are acknowledged as a complete nutrient for agriculture and have the potential to be utilized in foliar application to meet the nutritional requirements of plants. Here, foliar application of nutrients is a method of supplying essential nutrients to plants by spraying them directly onto the aerial part of plants (foliage), which includes leaves, stems, branches, etc. This approach is particularly useful when plants are experiencing nutrient deficiencies or when the soil conditions are not optimal for nutrient uptake. Thus, foliar application of nutrients offers several advantages over conventional soil application methods, which include direct supply of nutrients to the plants with rapid nutrient absorption through the leaves, efficient usage of nutrients for essential physiological processes with minimum nutrient wastage, and greater flexibility in nutrient management with minimal nutrient runoff and leaching into waterways contributing to pollution and eutrophication.
However, most of the contents of cow urine are generally disposed of as liquid waste in landfills or in waterbodies, thereby creating environmental issues such as the generation of ammonia in land and waterbodies, which causes foul odor in open fields if proper disposal procedures are not performed. However, to overcome these issues, cow urine is collected and transferred to the collection site for use as organic fertilizer on land farms. Through this, farmers can get extra income and also minimize the wastage of cow urine and its distillates from natural sources.
Cow urine and its distillates are natural, organic, inorganic, or synthetically fortified. Innovations in mineral nano-fortification, nano-sizing, and formulations based on cow urine have the potential to boost nutritional bioavailability and efficacy for productivity by a multifold increase. Biomolecules consist of various forms, such as natural organic or inorganic nutrients, that are required to complete the life cycle of plants. For the completion of a plant’s life cycle, different aqueous suspensions, such as cow urine and/or its distillates with nanoscale nutrients, are utilized for high agricultural production and other development prospects. Moreover, cow urine offers various advantages over the usage of chemical fertilizers. These advantages are:
1. Replacement of Water: The liquid formulation of a product containing >85% high-purity water are generally extracted from underground reservoirs are replaced with freely available cow urine or ark as a medium.
2. Environment pollution reduction: Cow urine contains urea as a major element which upon release in environment create water and air pollution thereby by utilizing the cow urine, significant reduction of pollution can be achieved.
3. Biodegradability: Cow urine is a natural, biodegradable substance that is environmentally friendly compared to synthetic dispersants and breaks down into harmless byproducts after the applicability thereby reducing the risk of environmental pollution.
4. Cost-Effectiveness: Cow urine is a cost-effective and readily available alternative to synthetic dispersants, making it a popular choice for farmers and agricultural practitioners in dairy and agricultural regions.
5. Nutrient Content: Cow urine contains essential nutrients and organic compounds that enhances the plant growth and productivity by providing additional nutrition sources.
Therefore, there is a need to provide a novel method for the preparation of nanostructured natural and hybrid composites of plant nutrient formulations using the distillation process of cow urine, i.e., “cow ark”. This “cow ark” is utilized as a natural dispersion medium for the application of nanofertilizers mixed with the ark either naturally or by fortifying with the required amount of macro- as well as micronutrients as per the need.
PRIOR ART AND ITS DISADVANTAGES:
The patent application no. CN1472176A relates to a biological compound fertilizer, in particular to a nano type active organic essence fertilizer and a preparation method thereof. A nano-class fermented active organic fertilizer is prepared from ammonium humate organonitrogen, ammonium polyphosphate, ammonium phosphate, calcium potassium pyrophosphate, organo potassium, additive and water through screening raw materials, grinding, biochemical reacting, adding additive, granulating, compounding, drying, cooling and coating. Its advantages are no poison and environmental pollution, and multiple functions including improving soil and crops, regulating growth of plant, increasing yield of crops, resisting diseases and bacteria, etc.
The major drawback of the said prior art is the non-usage of natural nutrients such as cow urine or its distillates as dispersing mediums for nanostructured natural and hybrid composite plant nutrients. Here, cited prior art provides a nano-class fermented active organic fertilizer prepared from synthetic substrates such as ammonium humate organonitrogen, ammonium polyphosphate, ammonium phosphate, calcium potassium pyrophosphate, etc. through screening raw materials, grinding, biochemical reactions, adding additives, granulating, compounding, drying, cooling, and coating. Whereas, the present invention utilizes natural components such as cow urine or its distillates as dispersing mediums for manufacturing nanostructured natural and hybrid composite plant nutrients.
The non-patent published paper titled “Effect of cow dung as organic manure on the growth, leaf biomass yield of Stevia rebaudiana and post-harvest soil fertility” represents an impact of organic manure such as cow dung on the growth, leaf biomass production, and post-harvest soil fertility of Stevia rebaudiana. Herein, a pot experiment was conducted to observe the effects of cow dung on growth, yield of stevia along with post-harvest fertility status of soil. Moreover, an overall performance of different forms of soils (non-calcareous soil and acid soil) were perceived wherein, non-calcareous soil was better than acid soil. Furthermore, all essential plant nutrients viz. total N, available P exchangeable K, Ca, Mg, available S, Zn, B and organic matter content of soil were significantly increased with the increased levels of Cow Dung in both the soils.
However, the said non-patentable prior art utilizes cow dung for the growth leaf biomass production, and post-harvest soil fertility of plant Stevia rebaudiana. Whereas, the present invention utilizes natural components such as cow urine or its distillates as dispersing mediums for manufacturing nanostructured natural and hybrid composite plant nutrients. In comparison to the present invention, the cited prior art does not use naturally structured nutrients in nano form, making it not economically feasible at the industrial level.
DISADVANTAGES OF THE PRIOR ART:
Most or all of the prior art and non-patent prior arts suffers from all or at least one of the following disadvantages:
• The prior art fails to provide a process for the preparation of nanostructured natural and hybrid composites of plant nutrient formulations.
• The prior art fails to provide a process for the preparation of nanostructured natural and hybrid composites of plant nutrient formulations that requires cow urine and/or its distillate as dispersing agent.
• The prior art fails to provide a process for the preparation of nanostructured natural and hybrid composites of plant nutrient formulations that utilize a natural nutrient-rich and/or fortified dispersion medium having nanoscale (l – 1000 nm) particles of the desired element and/or carrier and/or biomolecule for providing balanced nutrient and protection for the growth of plants.
• The prior art fails to provide a process for the preparation of nanostructured natural and hybrid composites of plant nutrient formulations that reduces the wastage of urine and is an extremely useful dispersion medium for the application of fertilizers, stimulants, nano formulations, etc.
• The prior art fails to provide a process for the preparation of nanostructured natural and hybrid composites of plant nutrient formulations that reduces the dependency on synthetic fertilizer.
• The prior art fails to provide a process for the preparation of nanostructured natural and hybrid composites of plant nutrient formulations that provides formulations to replenish the natural ecosystem.
• The prior art fails to provide a process for the preparation of nanostructured natural and hybrid composites of plant nutrient formulations that provide all the necessary macro- and micronutrients to plants encapsulated in natural and/or synthetic polymers in their required dosage and rate as per the standard practices followed worldwide.
• The prior art fails to provide a process for the preparation of nanostructured natural and hybrid composites of plant nutrient formulations that reduces the dependency fully and/or partially on fresh water, which acts as a dispersion medium for nano-formulation used for foliar application.
• The prior art fails to provide a process for the preparation of nanostructured natural and hybrid composites of plant nutrient formulations that reduces and converts the waste by-product into a viable product.
• The prior art fails to provide a process for the preparation of nanostructured natural and hybrid composites of plant nutrient formulations that reduces and converts the waste by-product into a growth-stimulating product.
• The prior art fails to provide a process for the preparation of nanostructured natural and hybrid composites of plant nutrient formulations that is ecologically friendly in nature.
• The prior art fails to provide a process for the preparation of nanostructured natural and hybrid composites of plant nutrient formulations that is economically feasible at the industrial level.

Thus, there is an unmet need to develop an invention that suffices the purpose of providing a novel manufacturing process of nano structured natural and hybrid composite of plant nutrient formulation using cow urine and/or its distillate as dispersing medium.
OBJECTS OF THE INVENTION:
Accordingly, the primary object of the present invention is to provide a novel manufacturing process of nano structured natural and hybrid composite of plant nutrient formulation.
Yet another object of the present invention is to provide a process for preparation of nano structured natural and hybrid composite of plant nutrient formulation that requires cow urine and/or its distillate as dispersing agent.
Yet another object of the present invention is to provide a process for the preparation of a nanostructured natural and hybrid composite of plant nutrient formulation that utilize a natural nutrient-rich and/or fortified dispersion medium having nanoscale (l – 1000 nm) particles of the desired element and/or carrier and/or biomolecule for providing balanced nutrient and protection for the growth of plants.
Yet another object of the present invention is to provide a process for the preparation of a nanostructured natural and hybrid composite of plant nutrient formulations that reduces the wastage of urine and is an extremely useful dispersion medium for the application of fertilizers, stimulants, nano formulations, etc.
Yet another object of the present invention is to provide a process for the preparation of a nanostructured natural and hybrid composite of plant nutrient formulation that reduces the dependency on synthetic fertilizer or formulation to replenish the natural ecosystem.
Yet another object of the present invention is to provide a process for the preparation of a nanostructured natural and hybrid composite of plant nutrient formulation that provide all the necessary macro- and micronutrients to plants encapsulated in natural and/or synthetic polymers in their required dosage and rate as per the standard practices followed worldwide.
Yet another object of the present invention is to provide a process for the preparation of a nanostructured natural and hybrid composite of plant nutrient formulation that reduces the dependency fully and/or partially on fresh water, which acts as a dispersion medium for nano-formulation used for foliar application.
Yet another object of the present invention provides a process for the preparation of a nanostructured natural and hybrid composite of plant nutrient formulation that reduces and converts the waste by-product into a viable product.
Yet another object of the present invention provides a process for the preparation of a nanostructured natural and hybrid composite of plant nutrient formulation that reduces and converts the waste by-product into a growth-stimulating product.
Yet another object of the present invention provides a process for the preparation of a nanostructured natural and hybrid composite of plant nutrient formulation that is ecologically friendly in nature.
Yet another object of the present invention provides a process for the preparation of a nanostructured natural and hybrid composite of plant nutrient formulation that uses green technology.
Yet another object of the present invention provides a process for the preparation of a nanostructured natural and hybrid composite of plant nutrient formulation that is economically feasible at the industrial level.
BRIEF DESCRIPTION OF DRAWING:
Fig. 1 : Shows the schematic diagram of the process for the preparation of the present invention.
Fig. 2 : Shows the electron micrograph of TEM based particle size analysis for natural nano NP
Fig. 3 : Graph showing DLS based particle size distribution analysis of natural nano NP
Fig. 4 : Graph showing zeta potential distribution analysis of natural nano NP

Meaning of reference numeral of the said component parts of the present invention to provide a process for the preparation of nano structured natural and hybrid composite of plant nutrient formulation:
D : Distillation
C : Cow
1 : Natural plant nutrient source
2 : Cow ark
3 : Ark + Natural plant nutrient
4 : Natural polymer
5 : Ark + Natural plant nutrient + Natural polymer
6 : High pressure homogenizer or Sonication
7 : Final product
8 : Commercially available cow urine

SUMMARY OF THE INVENTION
The present invention provides a novel manufacturing process of nano structured natural and hybrid composite plant nutrients and/or biomolecules using natural and/or synthetic polymers as an encapsulating agent. Moreover, the present invention provides a novel manufacturing a process for preparation of nano structured natural and hybrid composite of plant nutrient formulation that requires cow urine and/or its distillate as dispersing agent. The synthesized novel nano formulation reduces the dependency on natural fresh fully and/or partially which acts as a dispersion medium for nano-formulation used for foliar application as well as provide essential macro and micro nutrients to plants. The present invention reduces the dependency on using commercially available synthetic fertilizer to provide nutrient to plants for optimum growth and development. Furthermore, the present invention enhances the quality of plant production using naturally available nutrients and / or fortified nutrients through novel nano-structured formulation. Moreover, the present invention involves a synthesized process with eco-friendly formulation using green chemistry as well as to reduce the dependency of large consumption of the fertilizers.
DETAILED DESCRIPTION OF THE INVENTION
The present invention provides a novel manufacturing process of nano structured natural and hybrid composite plant nutrients and /or biomolecules using natural polymers as an encapsulating agent. Here, the raw materials utilized for the synthesis of nanostructured formulations include amino acids in single or combined nutrients as active natural sources of nitrogen and rock phosphate nutrients as active natural sources of phosphorus.
In particular, the present invention provides a novel manufacturing process for the preparation of nanostructured natural and hybrid composites of plant nutrient formulations that require cow urine and/or its distillate as dispersing agents. Here, cow ark or cow urine is utilized as a natural nutrient rich source having nitrogen as well as phosphorus for its formulation in said invention. In natural conditions, cow urine having adequate amount of nitrogen and phosphorus are not sufficient for the development of plants; thereby admixture of active nutrients such as nitrogen and phosphorus from organic sources are utilized along with cow ark as dispersing mixture for fortifying the nanoformulation and providing proper development to the plants. Moreover, the present invention provides an effective method of synthesizing a novel nano formulation that reduces the dependency on natural freshwater fully and/or partially and acts as a dispersion medium for nano-formulation used for foliar application as well as providing essential macro and micronutrients (currently 17 elements) for proper growth and development of plants.
Essential macronutrients primarily include Nitrogen (N), Phosphorus (P), Potash (K), Calcium (Ca), Sulfur (S), and Magnesium (Mg) in larger quantity requirement whereas, essential micronutrients such as Iron (Fe), Zinc (Zn), Copper (Cu), Boron (B), Manganese (Mn), Molybdenum (Mb), Chloride (Cl), and others are required in smaller quantity. In reference to worldwide acceptance, essential macro- and micronutrients vary on the basis of external environmental factors, including crop usage, soil, plant growth stage, etc. However, the standard NPK ratio accepted worldwide is 4:2:1, which improves soil health and increases crop yields in bulk when directly applied to the soil. With reference to the same, Nitrogen (N) and Phosphorus (P) are the two most important macronutrients, which play a plethora of roles in the life cycle of plants in terms of their growth and development. Here, N and P present in said invention fall in the ratio of 1:1, as these elements are applied directly to the leaves as a foliar application.
A healthy plant requires three key macronutrients: nitrogen (N), phosphorus (P), and potassium (K). Nitrogen is a necessary component of proteins for stem, leaf, and root growth; Phosphorus aids in the development of strong roots; and Potassium is required for tuber and fruit growth. Crop-livestock integration promotes nutrient cycling for sustainable agriculture. Cow urine has a wide range of agricultural applications. The custom stretches back to the Vedic era. It is a nutrient-rich natural resource that promotes soil fertility and crop productivity while encouraging agricultural sustainability through direct or indirect use. Except for small-scale practical applications, developing technologies for cow urine utilization have been under development for several decades due to their significant mineral and nutritional composition.
Here, said elements utilized are either naturally available or synthetically procured from authorized dealers, which imparts functioning in the form of either inhibiting at low concentrations or promoting biological function at optimum concentrations as plant nutrients, e.g., macro- and micronutrients. The present invention has the direct implication of drastically reducing the dosage of fertilizers, either through macro or micronutrients in the form of individual or mixed (hybrid) forms, as well as conserving the natural groundwater as a dispersing medium while increasing the efficiency of the nutrients supplied to plants.
According to the embodiments herein, urine (from cows, cattle, and/or other mammals), urine distillate form “ark” are procured from commercially available parties wherein distillate form of cow urine is obtained through direct complete distillation of cow urine (100%) and said distillation process of urine is taken through standard certified practices. Here, with respect to the quantity of cow urine admixed with freshwater is represented in tabular format thereby showcasing amount of cow urine admixed with fresh water required for the present invention.
Fresh water (%) Cow Urine (%)
1-25 99-75
25-50 75-50
50-75 50-25
75-99 25-1

According to the embodiments herein, plant nutrients or fertilizers refer to macronutrients, which include the group of nitrogenous fertilizer (amino acids and its chelates in single and/or in combination or any other bio-molecules having nitrogen content in the range of 1-40%), phosphoric fertilizer (mushroom compost, hair, rock phosphate, bone meal, burned cucumber skins, bat guano, fish meal, cottonseed meal, worm castings, blood meal, manure, and compost having phosphorus content in the range of 0.2–50%), potash, and/or other essential thirteen nutrients in combination, which may be of naturally occurring bio-molecules and/or deposits or sediments, compost, manure, etc., having the desired nutrient content. In terms of nitrogen (N) as active nutrient, most preferred nutrient sources are Acidic, basic and neutral side chain amino acids alone or in combination and its chelated form, sea weed etc. having nitrogen content in the range of 1-40% whereas for phosphorus (P) as active nutrient, most preferred nutrient sources are mushroom compost, hair, rock phosphate, bone meal, burned cucumber skins, bat guano, fish meal, cottonseed meal, worm castings, blood meal, manure, and compost having phosphorus content in the range of 0.2–50%.
Synthetic and/or organic forms of commercially available fertilizers that have all the necessary mineral nutrients, like macro- and micronutrients, which is used for the synthesis of cow urine or distillate-based nano formulations.
According to the embodiments herein, natural bio-polymers such as agar, starches, alginates, chitosan, cellulose, xanthan gum, Arabic gum, guar gum, or any other similar natural polymer capable of providing polymerization. In terms of natural bio-polymers, most preferred nutrient sources are cellulose and chitosan.
An embodiment for the process of nano structured natural and hybrid composite of plant nutrient formulation that employs cow urine or its distillates as a dispersing medium as illustrated in Figure 1 comprises of following steps:
i. collecting 500-1000 ml raw cow urine packed in plastic bottle from commercially available manufacturers in beaker,
ii. filtrating said cow urine in Whatman No. 1 filter paper having a pore size of 25 µm for the removal of large sized impurities, debris, or precipitated materials through conventional filtration and sedimentation processes,
iii. carrying out the distillation process of filtered cow urine obtained from step (ii) to obtain 650–750 ml of cow ark as distillate form of the available cow urine (1000 ml) and / or commercially available cow ark
iv. preparing concentrated aqueous solution of natural plant nutrients such as amino acids in single or combined form, rock phosphate etc. by dissolving said natural plant nutrient in filtered and distillated form of cow urine (cow ark) and followed by agitation and stirring for particular time intervals with a regular speed interval in the form of rpm under standard room temperature conditions for complete dissolution,
v. adding and mixing natural polymer as stabilizing and capping agent with concentrated aqueous solution of natural plant nutrients (as output from step iv) for obtaining polymerized form of cow ark mixture at an appropriate temperature condition ranging from 28 to 38°C in beaker,
vi. substantially, preparing a homogenous solution of said mixture (as output from step v) through a high-pressure homogenizer with an optimal pressure ranging from 1-1000 bar and passing through each pressure for 1-3 times with a feeding flow rate of 8–10 L/hr depending upon the reduction of particle size at an appropriate temperature condition ranging from 28 to 38°C in the beaker, and
vii. obtaining 1000ml resulting solution of colloidal dispersion urine or ark-based plant nutrient nanoparticles as final end product.
The first step in an embodiment of the present invention is to collect 500-1000 mL of raw cow urine packed in plastic bottle from commercially available manufacturers in beaker.
The second step of an embodiment is to filter out said commercially available cow urine obtained from step (i) by using Whatman No.1 filter paper with a pore size of 25 µm through conventional filtration and sedimentation processes for the removal of large-sized impurities, debris, or precipitated materials present in said cow urine.
The third step in an embodiment of the present invention is to carry out the distillation process of filtered cow urine from step (ii) for obtaining 650-750 ml of cow ark as distillate form of 1000ml available cow urine. At the end of the process, approximately 65-75% of the cow ark is recovered after carrying out the distillation process of cow urine.
Herein, distillate water can also be used for nanoparticles formulation as a dispersion medium when there is less or no supply of cow urine or ark. Thus, formulation of specific macro- or micronutrients using natural and/or organic sources of nutrients are not limited to the distillation of available cow urine.
The fourth step in an embodiment of the present invention is to prepare a concentrated aqueous solution of plant nutrients containing nitrogen (N) in the range of 1-5% and phosphorus (P) in range of 1-8% by dissolving said solution in the filtered or distillate urine, followed by agitation and/or stirring for at least 10, 20, 30, 45, 60, or 120 minutes at 100 to 200, 200 to 300, or 300 to 400 rpm under laboratory standard room temperature conditions ranging from 28 to 38°C until the fortified or added plant nutrients solubilize or dissolve completely. Here, agitation is performed to allow optimum mixing of the raw materials, thereby allowing maximum interaction between the opposite charge entities for their bonding and the formation of stable nanostructures.
The fifth step in an embodiment of the present invention is to add and mix natural polymer mixture as stabilizing and capping agent with concentrated aqueous solution of natural plant nutrients (as output from step iv) for obtaining polymerized form of cow ark mixture in an appropriate temperature condition ranging from 28 to 38°C followed by addition of pH stabilizing agent.
The sixth step in an embodiment of the present invention is to prepare a homogenous solution of said mixture (as output from step v) through high pressure homogenizer having optimal pressure ranging from 1-1000 bar (1-100, 100-200, 200-300, 300-400, 400-500, and 500-1000 bar) and passing through each pressure for 1-3 times with a feeding flow rate of 8–10 L/hour depending upon the reduction of particle size at an appropriate temperature condition ranging from 28 to 38°C in the beaker and / or through sonication process.
The seventh and final step in an embodiment of the present invention is to formulate resulting solution of colloidal dispersion urine or ark-based plant nutrient nanoparticles as final end product having pH ranging between 6.5-8.5.
Further physical and chemical properties of said resulting solution of colloidal dispersion urine or ark-based plant nutrient nanoparticles for providing a suitable formulation for balanced plant growth and development are enlisted below in TABLE 1.
Sr. No. Property Result for nanoformulation
1 Form Liquid
2 Colour Transparent to light yellowish
3 Total Nitrogen (percent by weight) 1.0-2.0
4 Total Phosphorous (as P2O5) percent by weight 1.0-8.0
5 pH (Liquid) 6.5-8.5
6 Viscosity 18.20
7 Particle size by TEM 34.45 nm
8 Particle size by DLS <90 nm
9 Surface charge / zeta potential (+/- scale) -15.0
10 Solubility in water Yes
11 Changes in physical state No

A tabular format showcasing the role of certain chemical or agents utilized in the process of nano structured natural and hybrid composite of plant nutrient formulation is represented in TABLE 2:
Sr No Name of Chemical Role Examples used in present invention Stages during the process
1 Acidic, basic and neutral side chain amino acids alone or in combination, amino acid chelates and sea weed Active ingredient and source of N Aspartic Acid, Glutamic Acid, Arginine, Histidine, Threonine, Glycine, Alanine alone or in combination and with metal chelates. Sea weed can also be utilized. Step iv
2 Rock phosphate Active ingredient and source of P Rock phosphate Step iv
3 Natural bio-polymer Stabilizing or capping agent Cellulose and chitosan Step v
3 Water soluble cationic polymeric material Capping agent Cellulose and chitosan and its oligomers, Step v
4 Water soluble anionic polymeric material Slow releasing agent Alginates, pectin, guar gum, xanthan gum Step v
5 pH stabilizing agent pH stabilization Citric acid Step vi
6 Preservative agent Preservation during the storage Thyme, citric acid alone Step vi

Said present invention is a simple process for the synthesis of nano plant nutrients using naturally available dispersing medium based on cow urine or its distillate form, 'ark', resulting in an environmentally and plant-friendly procedure. Furthermore, said invention utilizes naturally occurring dispersing medium as well as precursors and polymers of natural origin, thus, saving natural groundwater as well as energy. Herein, energy is referred to electric energy employed for collecting water from the underground and utilized for the purification of groundwater to attain distilled water.
WORKING EXAMPLES OF THE INVENTION:
Maize is known as one of the world’s leading crop is widely cultivated as cereal grain in India and most primarily acknowledged as queen of cereals due of its highest genetic yield potential by individuals, thereby, maize crop is considered as third important food crop after rice and wheat in regards to nutrient consumption.
Various field experiments were conducted at Anand Agricultural University, Anand, Gujarat to study the comparative analysis between nano-structured and bulk fertilizer treatment in maize crop for their growth and biochemical attributes. These experiments were laid out in a Randomized Complete Block Design as illustrated in Tables 3-6, with 6 treatments (T1, T2, T3, T4, T5 and T6). Herein, various aspects of the study for their morpho-physiological parameters along with their biochemical attributes were recorded as tabular data with reference to dual spraying of Nano-DAP among treatments over control and fertilizer application either with 50% or 75% were recorded at 30-35 and 50-55 days after sowing for their spray efficiency estimation. The spraying was done with the help of knapsack sprayer having flat fan nozzle with a spray fluid of 500 L/ha.
These treatments were:
T1: Control treatment (no fertilizer treatment)
T2: 100% Recommended Dosage of Nitrogen and Phosphorous (RDNP)
T3: 75% Recommended Dosage of Nitrogen and Phosphorous (RDNP)
T4: 50% Recommended Dosage of Nitrogen and Phosphorous (RDNP)
T5: Addition of 4mL/L Natural nano NP formulation (NP2) with T3 treatment as foliar spray
T6: Addition of 4mL/L Natural nano NP formulation (NP2) with T4 treatment as foliar spray
TABLE 3: Comparative analysis between nanostructured and bulk fertilizer in maize over control treatment and 75% of fertilizer application with respect to their morpho-physiological parameters.
Sr. no. Growth and Yield Parameters Comparison among treatments with respect to spray efficiency of dual spraying of Natural nano NP formulation over control treatment and 75% of fertilizer application
T3 (%) increase over control T5 (%) increase over control T5 (%) increase over respective T3 concentration
1. Plant height at harvest 18.94 52.89 28.54
2. LAI at 60 DAS 28.29 68.29 31.18
3. No. of cobs/plant 10.57 39.02 25.74
4. Cob length (cm) 11.77 40.34 25.56
5. Cob weight (g) 23.33 85.38 50.31
6. Cob yield (kg/ha) 8.52 34.91 24.32
7. Grain yield (kg/ha) 51.38 80.10 18.97
8. Straw yield (kg/ha) 50.09 78.62 19.01
9. Seed index (g) 18.98 38.16 16.13
TABLE 4: Comparative analysis between nanostructured and bulk fertilizer in maize over control treatment and 50% of fertilizer application with respect to their morpho- physiological parameters.
Sr. no. Growth and Yield Parameters Comparison among treatments with respect to spray efficiency of dual spraying of Natural nano NP formulation over control treatment and 50% of fertilizer application
T3 (%) increase over control T5 (%) increase over control T5 (%) increase over respective T3 concentration
1. Plant height at harvest 10.33 28.29 16.27
2. LAI at 60 DAS 17.56 38.54 17.84
3. No. of cobs/plant 8.94 20.33 10.45
4. Cob length (cm) 10.23 24.35 12.81
5. Cob weight (g) 8.28 52.88 41.18
6. Cob yield (kg/ha) 4.68 15.91 10.73
7. Grain yield (kg/ha) 49.23 65.29 10.76
8. Straw yield (kg/ha) 43.11 62.77 13.74
9. Seed index (g) 17.81 25.67 6.68

TABLE 5: Comparative analysis between nanostructured and bulk fertilizer in maize over control treatment and 75% of fertilizer application with respect to their biochemical attributes.
Sr. no. Biochemical attributes Comparison among treatments with respect to spray efficiency of dual spraying of Natural nano NP formulation over control treatment and 75% of fertilizer application
T3 (%) increase over control T5 (%) increase over control T5 (%) increase over respective T3 concentration
1. Chlorophyll a
(mg/gm) 5.66 44.30 36.56
2. Chlorophyll b
(mg/gm) 51.87 123.36 47.08
3. Total Chlorophyll (mg/gm) 12.32 55.69 38.61
4. Total soluble Sugars (%) 2.27 22.73 20.00
5. Phenol (%) 2.56 42.31 38.75
6. Reducing sugars (%) 5.17 39.66 32.79
7. Crude protein (%) 5.25 9.17 3.72
8. Crude fiber (%) 1.32 3.51 2.16
9. Starch (%) 0.66 3.70 3.02
TABLE 6: Comparative analysis between nanostructured and bulk fertilizer in maize over control treatment and 50% of fertilizer application with respect to their biochemical attributes.
Sr. no. Biochemical attributes Comparison among treatments with respect to spray efficiency of dual spraying of Natural nano NP formulation over control treatment and 50% of fertilizer application
T3 (%) increase over control T5 (%) increase over control T5 (%) increase over respective T3 concentration
1. Chlorophyll a
(mg/gm) 4.80 44.30 9.98
2. Chlorophyll b
(mg/gm) 43.46 123.36 13.36
3. Total Chlorophyll (mg/gm) 10.37 55.69 10.62
4. Total soluble Sugars (%) 0.45 22.73 14.93
5. Phenol (%) 15.38 42.31 13.33
6. Reducing sugars (%) 5.17 39.66 9.84
7. Crude protein (%) 1.96 9.17 3.84
8. Crude fiber (%) 2.63 3.51 -1.71
9. Starch (%) 0.92 3.70 1.19

For morpho- physiological observation:
As illustrated in tabular format, Tables 3-4 represent comparative analysis table between nanostructured and bulk fertilizer in maize over control treatment and fertilizer application of 75% and 50% with respect to their morpho- physiological parameters showcasing their optimal growth and yield of plants. Here, in each net plot area, five plants were randomly selected for recording the morpho-physiological observations such as plant height at harvesting stage, leaf area index (LAI), number of cobs per plant, cob length, cob weight, cob yield, seed index, grain and straw yield.
For biochemical observation:
As illustrated in tabular format, Tables 5-6 represent comparative analysis table between nanostructured and bulk fertilizer in maize over control treatment and fertilizer application of 75% and 50% with respect to their biochemical attributes including chlorophyll content, total soluble sugars, total phenol, reducing sugars, crude protein, crude fiber and starch content. Here, biochemical attributes are carried out as per the methodology depicted in literature studies as standard analytical reference method.
From the above-mentioned tables (Tables 3-6), said invention facilitated an experimental study on the impact of nano-NP on nutrient-demanding maize crops, wherein conventional urea and DAP application with a reduced dosage of fertilizer application of 75% and 50% is combined with dual spray application of novel nano-NP over 100% conventional fertilizer application, thereby, providing a similar exhibition of optimal growth and development of maize crops. Thus, from overall observation and results, it can be concluded that said invention, having a reduced dosage of nano-NP foliar spraying treatment, exhibits similar morpho-physiological and biochemical attributes as compared to conventional bulk synthetic fertilizer treatments, leading to no negotiation on the economic potential of crops required for technology acceptance by the farmers.
TABLE 7: Effect of natural nano-NP application on the total microbial count in soil of maize crops.
Treatments Soil microbial count (cfu/gm)
Location 1 Location 2 Pooled
Initial estimation 5.72
(5.2 x 105) 5.97
(9.4 x 105) -
T1 6.64
(4.4 x 106) 6.95
(8.8 x 106) 6.79
(6.6 x 106)
T2 6.46
(2.9x 106) 6.95
(8.6x 106) 6.69
(5.7 x 106)
T3 6.67
(4.8 x 106) 6.98
(9.5 x 106) 6.82
(7.1 x 106)
T4 6.69
(5.0 x 106) 6.99
(9.9 x 106) 6.84
(7.4 x 106)
T5 6.66
(4.6 x 106) 6.98
(9.5 x 106) 6.82
(7.0 x 106)
T6 6.68
(4.9 x 106) 6.98
(9.5 x 106) 6.83
(7.2 x 106)
cfu = colony forming unit
An embodiment referring to Table 7 shows the microbial population recorded for each treatment (T1–T6), which highlights the effect of natural nano-NP application as a foliar spray fertilizer on the total microbial count in the soil of maize crops. Here, a negative impact is observed on the soil microbial population for chemical fertilizer when it is directly applied to the soil surface. These microbial populations include beneficial organisms, which play an important role in the overall plant response to abiotic and biotic stresses. Thus, there is a major worldwide concern about soil degradation due to excessive usage of chemical fertilizers, which hampers the overall production and productivity of crops. To overcome the soil degradation issue, foliar application of nano NP is directly applied to the plant canopy through foliar spraying, thereby showcasing no negative impact on soil microbes when directly exposed to plants. Thus, the foliar application of nano NP directly results in the improvement of soil as well as better crop response to fertilizer application and resilience towards abiotic and biotic stresses.
In another embodiment, an image from Transmission electron microscopy (TEM) as illustrated in figure 2 has been recorded for natural nano NP sample formulation. Here, particle size analysis of natural nano NP formulation sample is determined to be 34.45 nm approximately through TEM microscopic analysis.
In an embodiment of present invention, figure 3 and figure 4 represents the graphical illustrations on particle size distribution and zeta potential distribution for formulation of nanostructured NP fertilizer respectively. The measurement of particle size potential value distribution and zeta potential value distribution for nano-NP dispersion was carried out using Particle Size Analyzer. Here, zeta potential distribution of nano NP was determined to be -15.0 mV according to the Smoluchowski equation while the nanoparticle size distribution was measured using the dynamic light scattering (DLS) method. Here, the hydro-dynamic diameter of particles was determined to be <90 nm using diffusion coefficients of Stoke-Einstein equation. For a poly-dispersed suspension, the obtained mean particle size called ‘z-average’ diameter (dz) is determined by the intensity of the light scattered by the particles and through DLS method, measurements were conducted at 25°C to obtain the z-average particle size ranging from 2 nm to 6 µm with polydispersity index of 0.29.
For zeta potential measurements, electrokinetic potential is specified for a colloidal system, which is a physical property exhibited by any particle in a suspension, and the magnitude of the zeta potential offers the net charge at the diffuse boundary of a particle in a suspension, which ultimately indicates the potential stability of a colloidal system. Hence, if the particles in a suspension have large negative or positive zeta potential values, particles will repel each other with no aggregation of nanoparticles. On the other hand, if particles have small zeta potential values, they will exhibit mutual attraction towards each other and form aggregates of nanoparticles without any external force to keep them apart.
Various safety studies, such as cytotoxicity safety studies, genotoxicity safety studies, and environmental safety studies, were performed in which the formulation of nanostructured NP liquid fertilizer successfully passed all their safety tests as per the OECD or SOP guidelines at an NABL-certified laboratory. Thus, said formulation of nanostructured NP liquid fertilizer is safer for the environment with a good significance of non-toxicity level.
ADVANTAGES OF THE INVENTION:
The present invention provides a process for the preparation of nano structured natural and hybrid composite of plant nutrient formulation.
The present invention provides a process for the preparation of nano structured natural and hybrid composite of plant nutrient formulation that requires cow urine and/or its distillate as dispersing agent.
The present invention provides a process for the preparation of nano structured natural and hybrid composite of plant nutrient formulation that utilizes a natural nutrient-rich and/or fortified dispersion medium having nanoscale (l – 1000 nm) particles of the desired element and/or carrier and/or biomolecule for providing balanced nutrient and protection for the growth of plants.
The present invention provides a process for the preparation of nano structured natural and hybrid composite of plant nutrient formulation that reduces the wastage of urine and is an extremely useful dispersion medium for the application of fertilizers, stimulants, nano formulations, etc.
The present invention provides a process for the preparation of nano structured natural and hybrid composite of plant nutrient formulation that reduces the dependency on synthetic fertilizer or formulation to replenish the natural ecosystem.
The present invention provides a process for the preparation of nano structured natural and hybrid composite of plant nutrient formulation that provide all the necessary macro- and micronutrients to plants encapsulated in natural and/or synthetic polymers in their required dosage and rate as per the standard practices followed worldwide.
The present invention provides a process for the preparation of nano structured natural and hybrid composite of plant nutrient formulation that reduces the dependency fully and/or partially on fresh water, which acts as a dispersion medium for nano-formulation used for foliar application, thereby saving lots of natural ground water.
The present invention provides a process for the preparation of nano structured natural and hybrid composite of plant nutrient formulation that reduces and converts the waste by-product into a viable product.
The present invention provides a process for the preparation of nano structured natural and hybrid composite of plant nutrient formulation that reduces and converts the waste by-product into a growth-stimulating product.
The present invention provides a process for the preparation of nano structured natural and hybrid composite of plant nutrient formulation that is energy efficient.
The present invention provides a process for the preparation of nano structured natural and hybrid composite of plant nutrient formulation that is ecologically friendly in nature.
The present invention provides a process for the preparation of nano structured natural and hybrid composite of plant nutrient formulation that uses green technology
The present invention provides a process for the preparation of nano structured natural and hybrid composite of plant nutrient formulation that is economically feasible at the industrial level.

,CLAIMS:We claim,

1. A process for preparation of nano structured natural and hybrid composite plant nutrients comprises of:
i. collecting commercially available cow urine and/or cow ark and preparing concentrated aqueous solution of natural plant nutrients containing nitrogen and phosphorus constituents by dissolving said natural plant nutrient in collected cow urine and/or cow ark, followed by agitation and stirring for particular time intervals of 10-120 minutes with a regular speed interval of 100-400 rpm under standard room temperature conditions ranging from 28-38 °C for complete dissolution,
ii. adding and mixing natural polymer as stabilizing and capping agent with concentrated aqueous solution of natural plant nutrients obtained from step (i), and obtaining polymerized form of cow ark mixture at an appropriate temperature condition ranging from 28 to 38°C in beaker,
iii. substantially, preparing a homogenous solution of said mixture obtained from step (ii) through a high-pressure homogenizer with an optimal pressure ranging from 1-1000 bar and passing through each pressure for 1-3 times with a feeding flow rate of 8–10 L/hour depending upon the reduction of particle size at an appropriate temperature condition ranging from 28 to 38°C in the beaker, and
iv. obtaining resulting solution of colloidal dispersion urine or ark-based plant nutrient nanoparticles as final end product through high pressure homogenizer and sonicator at a specific pH level ranging from 6.5-8.5.

2. The process for preparation of nano structured natural and hybrid composite plant nutrients as claimed in claim 1, wherein said concentrated aqueous solution of natural plant nutrients are selected from group comprising nitrogenous fertilizer (arginine and/or any other bio-molecules having nitrogen content in the range of 1-40%), phosphoric fertilizer (mushroom compost, hair, rock phosphate, bone meal, burned cucumber skins, bat guano, fish meal, cottonseed meal, worm castings, blood meal, manure, and compost having phosphorus content in the range of 0.2–50%), potash, and other essential nutrients in combination of naturally occurring bio-molecules, deposits, sediments, compost or manure.

3. The process for preparation of nano structured natural and hybrid composite plant nutrients as claimed in claims 1 and 2, wherein selected concentrated aqueous solution of natural plant nutrients having nitrogen constituents are amino acids in single or combined form having acidic, basic and neutral side chain, amino acid chelates and sea weed in range of 1-5%.

4. The process for preparation of nano structured natural and hybrid composite plant nutrients as claimed in claim 1, wherein selected concentrated aqueous solution of natural plant nutrients having phosphorus constituents is rock phosphate in range of 1-8%.

5. The process for preparation of nano structured natural and hybrid composite plant nutrients as claimed in claim 1, wherein said natural polymer selected from group comprising agar, starches, alginates, chitosan, cellulose, xanthan gum, Arabic gum, guar gum.

6. The process for preparation of nano structured natural and hybrid composite plant nutrients as claimed in claims 1 and 5, wherein selected natural polymer are cellulose and chitosan.

7. The process for preparation of nano structured natural and hybrid composite plant nutrients as claimed in claim 1, wherein said pH stabilizing agent is citric acid.

Dated this 10th Day of May, 2024.

Gopi Jatin Trivedi (Ms)
IN/PA/993
Authorized Agent of Applicant
To,
The Controller of Patents,
The Patent Office,
At Mumbai.