Inhibitor inclusion complex nitrogen fertilizer compositions

Abstract

A fertilizer composition comprising a solidified nitrogen fertilizer melt, said melt comprising one or more cyclodextrin inhibitor inclusion complexes comprising at least one c cyclodextrin and / or derivative thereof complexed with an inhibitor, and methods for producing and using the same are disclosed. The inhibitor comprises at least one nitrification inhibitor and / or at least one urease inhibitor. The one or more cyclodextrin-inhibitor inclusion complexes is homogeneously dispersed throughout the solidified nitrogen fertilizer melt.

Description

INHIBITOR INCLUSION COMPLEX NITROGEN FERTILIZER COMPOSITIONS CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to and the benefit of European Patent Application No. EP23190387, filed on 08 August 2023. The contents of the referenced application are incorporated into the present application by reference.BACKGROUND OF THE INVENTIONA. Field of the Invention

[0002] This invention is in the field of controlled release fertilizer compositions. Generally, it concerns a nitrogen fertilizer composition that includes an inhibitor-inclusion complex dispersed throughout a solidified nitrogen fertilizer melt.B. Description of Related Art

[0003] Fertilizers are chemical compositions that are added to plants and crops in order to provide nutrients that promote growth. Nitrogen, phosphorus, and potassium, or NPK, are present in many fertilizers, and these three primary nutrients play key roles in plant nutrition and growth. Nitrogen is considered to be the most important nutrient, and plants absorb more nitrogen than any other element. Nitrogen is essential for the formation of proteins, and proteins make up a significant portion of most living tissues. The second primary nutrient, phosphorus, is linked to a plant’s ability to use and store energy, including the process of photosynthesis. Potassium is the third primary nutrient of commercial fertilizers. It helps strengthen resistance to disease and plays an important role in increasing crop yields and overall quality. Potassium also protects the plant when the weather is cold or dry, strengthening its root system and preventing wilt.

[0004] Fertilizers offer different release profiles in which the rate at which nitrogen, phosphorus, and potassium nutrients are released is variable. Most commercial fertilizers are water-soluble quick-release fertilizers (QRFs) that quickly release nutrients when placed in soil. These quick-release fertilizers can be impractical and / or unpredictable, especially when high-leaching events like overwatering or flooding occur. Controlled-release fertilizers (CRFs) are fertilizers that contain a plant nutrient in a form the plant cannot immediately absorb. Controlled-release fertilizers are typically coated or encapsulated with materials that control the rate, pattern, and duration of plant nutrient release. A number of fertilizer production techniques have been developed that include polymeric components in attempts toslow the release of fertilizer nutrients. Oftentimes, the encapsulating materials are not biodegradable and accumulate in soils and plants overtime.

[0005] Attempts have also been made to coat or mix fertilizers (e.g., urea fertilizers) with inhibitors such as nitrification and / or urease inhibitors. One of the issues with the coating and mixing processes is that the nitrification and / or urease inhibitors can be subjected to temperatures that can cause degradation and reduce the effectiveness of the inhibitors.SUMMARY OF THE INVENTION

[0006] A solution to at least one or more of the aforementioned problems has been discovered. In one aspect, the solution can include the formation of a fertilizer composition having a nitrogen fertilizer and a cyclodextrin-inhibitor inclusion complex. The inhibitor (e.g., a nitrification and / or urease inhibitor), when complexed with the cyclodextrin, can be thermally stable at temperatures (e.g., 140 °C or more) exceeding the melt temperature of the nitrogen fertilizer. This can be advantageous in that the complexes can be contacted with and dispersed throughout molten nitrogen fertilizer with reduced, minimal, or no degradation of the inhibitors. Once the molten nitrogen fertilizer is solidified (e.g., through cooling), the resulting fertilizer composition can include a solidified nitrogen fertilizer melt with one or more of the cyclodextrin-inhibitor inclusion complexes dispersed throughout the solidified melt. In one aspect, the fertilizer composition can include a continuous phase comprising the nitrogen fertilizer melt and a discontinuous phase comprising one or more (e.g., a plurality) of the cyclodextrin-inhibitor complexes dispersed throughout the continuous phase. Without wishing to be bound by theory, it is believed that complexing the inhibitor with the cyclodextrin results in thermal protection to the inhibitor such that it can be contacted with molten nitrogen fertilizer with minimal to no degradation of the inhibitor. Additionally, the molten nitrogen fertilizer can act like a solvent to the complexes such that individual molecules of the cyclodextrin-inhibitor inclusion complexes can be efficiently dispersed throughout the molten nitrogen fertilizer, which allows for substantially even or homogenous dispersion of the complexes throughout the molten nitrogen fertilizer; in some aspects, this can be analogous to NaCl being dissolved in water where individual molecules of Na+ and Cl- are evenly or homogenously dispersed throughout the continuous water phase with one difference — the cyclodextrin-inhibitor complex remains a complex but is otherwise dissolved into individual molecules of the complex within the molten nitrogen fertilizer. Once the molten nitrogen fertilizer is solidified, the individual molecules of the complexes remainevenly or homogenously dispersed throughout the continuous network of the solidified nitrogen fertilizer melt. The solidified nitrogen fertilizer melt can include any one of the fertilizers disclosed throughout the specification. An advantage of the fertilizer compositions of the present invention is that there is minimal to no segregation or coalescence of the cyclodextrin-inhibitor inclusion complexes with each other within the continuous network of solidified nitrogen fertilizer melt, which can result in efficient protection of the nitrogen during storage and / or use of the fertilizer composition. Still further, this efficient dispersion of molecules of the complexes throughout the continuous solidified network can allow for reduced amounts of inhibitor used to make the fertilizer composition.

[0007] A fertilizer composition is disclosed herein that employs a renewable and biodegradable complexing component, and an inhibitor component that is included within the complexing component to form an inhibitor inclusion complex. The inhibitor inclusion complex may be embedded in a nitrogen fertilizer. Upon exposure to moisture, at least part of the nitrogen fertilizer component may dissolve rapidly, thereby providing an immediate release of fertilizer nutrients. The inhibitor inclusion complex, by contrast, is less soluble and resists immediate solubilization, this allows the inhibitor component to release its fertilizer nutrients slowly over time. The resulting bi-phasic nutrient release profile provides both immediate and sustained nutrient release that is ideal for crop fertilization. The complexing component may spatially enclose the one or more inhibitor(s) in a inclusion complex. The complexing component may be a compound capable of forming an equilibrium with one or more inhibitor. The equilibrium may be a dynamic equilibrium where similar amounts of inhibitor are entering the spatial enclosure of the complexing component as are leaving the spatial enclosure of the complexing component. The complexing component may be a polymer, forming a polymer-inhibitor inclusion complex. The polymer may be a polysaccharide and / or oligomer such as a cellulose, a starch, a cyclodextrin, and / or a derivative thereof. The complexing component may be a cyclodextrin and / or a derivative thereof, forming a cyclodextrin-inhibitor inclusion complex.

[0008] Cyclodextrins (CyDs) can form an inclusion complexes with various molecules which are either hydrophobic or having hydrophobic tails. The advantage of the CyDs complexing with the said molecules is that, the guest molecules will not be fixed in the CyDs but rather they are in dynamic equilibrium; thereby, the guest molecule can be released slowly in an aqueous environment through triggering mechanism. Moreover, formedCyD / host complex can have several advantages like protecting the guest molecules from multiple external environmental factors like light, heat, biochemical degradation, and also improves the thermal stability, enhance solubility and reduce odor. However, the actual application of CyDs as hosts in agriculture is rarely reported. Here, we propose a cheaper and more straightforward formulation of the urease inhibitors (NBTPT, NPPT, and NBPT) and nitrification inhibitors (DCD and DMPP) using CyDs. The nitrification inhibitors prevent bacteria in the soil from converting the ammonium portion of N from the manure into nitrate and reduce the risk of nitrate leaching and denitrification using CyDs. The optimized CyDs encapsulated inhibitors are easy to mix with the urea melt and the thermally stable.

[0009] In one aspect, the fertilizer composition comprises a nitrogen fertilizer, and a cyclodextrin-inhibitor inclusion complex comprising at least one cyclodextrin and / or derivative thereof complexed with an inhibitor, wherein the inhibitor comprises at least one nitrification inhibitor and / or at least one urease inhibitor, and wherein at least one cyclodextrin-inhibitor inclusion complex is embedded in the nitrogen fertilizer. In another aspect, the fertilizer composition contains the cyclodextrin-inhibitor inclusion complex embedded in a solidified nitrogen fertilizer melt. In some aspects, the cyclodextrin and / or derivative thereof may comprises a-cyclodextrin, [3-cyclodextrin, and / or y-cyclodextrin. In some aspects, the cyclodextrin and / or derivative thereof comprises y-cyclodextrin.

[0010] In some aspects, the inhibitor inclusion complex may include a nitrification inhibitor.The nitrification inhibitor may be one or more of 3,4-dimethylpyrazole phosphate (DMPP), thio-urea (TU), dicyandiamide (DCD), 2-chloro-6-(trichloromethyl)pyridine (nitrapyrin), 5- ethoxy-3-trichloromethyl- 1 ,2,4-thiadiazol (TERRAZOLE™), 2-amino-4-chloro-6-methyl- pyrimidine (AM), 2-mercapto-benzothiazole (MBT), ammonium thiosulfate (ATS), 2- sulfanimalamidothiazole (ST), or a combination thereof.

[0011] In some aspects, the inhibitor inclusion complex may include a urease inhibitor. The urease inhibitor may be one or more of N-(n-butyl)-thiophosphoric triamide (NBTPT), N-(n- butyl)-phosphoric triamide, N-(n-propyl) thiophosphoric triamide, benzoylthiourea (BTU), hydroquinone, acetohydroxamic acid (AHA), hydroxyurea (HU), and / or phenyl phosphorodiamidate (PPDA). In some aspects, the urease inhibitor is N-(n-butyl)- thiophosphoric triamide (NBTPT).

[0012] In certain embodiments, the cyclodextrin-inhibitor inclusion complex comprises y- cyclodextrin or a derivative thereof and NBTPT. In certain embodiments, the solidified nitrogen fertilizer melt comprises urea, and the cyclodextrin-inhibitor inclusion complex, wherein the cyclodextrin-inhibitor inclusion complex comprises y-cyclodextrin or a derivative thereof and NBTPT. In certain embodiments, the molar ratio of y-cyclodextrin or a derivative thereofNBTPT in the cyclodextrin-inhibitor inclusion complex is 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, 5: 1, 4: 1, 3: 1, 2: 1, 1: 1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1: 10, or any range or value therebetween. In certain embodiments, the molar ratio of y-cyclodextrin or a derivative thereofNBTPT in the cyclodextrin-inhibitor inclusion complex is 2: 1, 1.5: 1, 1: 1, 1: 1.5 or 1:2, or any range or value therebetween. In certain embodiments, the molar ratio of y-cyclodextrin or a derivative thereofNBTPT in the cyclodextrin-inhibitor inclusion complex is 10: 1 to 1: 10; 5: 1 to 1:5; 4: 1 to 1:4; 2: 1 to 1:2. In certain embodiments, the molar ratio of y- cyclodextrin or a derivative thereofNBTPT in the cyclodextrin-inhibitor inclusion complex is 2: 1 to 1:2.

[0013] In some aspects, the inhibitor inclusion complex may include both a nitrification inhibitor and a urease inhibitor, such as N-(n-butyl)-phosphoric triamide and dicyandiamide. Some commercial sources of inhibitors and combination inhibitors include, but are not limited to N-SERVE (BASF), NITROGARD (YARA), NITRIFICIN (NOVOZYMES), NITROGRAD PLUS (YARA), NITROGEN FIXER PLUS (AGRI LIFE RESEARCH), UREA STABILIZER (PLANT FOOD TECHNOLOGY), UREAPLUS (BIOMIN), and NITROGNEX (A M. CASTLE).

[0014] In some aspects, the inhibitor inclusion complex is thermally stable at 130 °C or more, such as 140 °C, or at a temperature of any one of, less than, greater than, between, or any range thereof of 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143,144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161,162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179,180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197,198, 199, and 200 °C. In some aspects, the inhibitor inclusion complex is stable at a pH range between 4 to 11, such as 5 to 10, or any one of, less than, greater than, between, or any range thereof of 4, 5, 6, 7, 8, 9, 10, and 11.

[0015] One aspect of the present invention is directed to the fertilizer composition. The fertilizer composition can contain 80 to 99.9999 wt. % of the nitrogen fertilizer, and / or0.0001 to 15 wt.% of the inhibitor inclusion complex. In certain aspects, the composition may include at least any one of, at most any one of, equal to any one of, or between any two of 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 wt.% of nitrogen fertilizer. In certain aspects, the composition may include at least any one of, at most any one of, equal to any one of, or between any two of 0.0001, 0.0005, 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 wt. % of inhibitor inclusion complex. In some aspects, the fertilizer composition further comprises a wetting agent, a carrier, a fdler agent, and / or a binder. In one aspect, the nitrogen fertilizer comprises urea. In certain aspects, the urea comprises a solidified urea melt. In some aspects, the fertilizer composition is homogenous.

[0016] In some aspects, the fertilizer composition comprises a ratio of inhibitor to degradation products of the inhibitor of 80:20 to 100:0. In some aspects, the ratio is, is at least, is at most, or is between 80:20, 85: 15, 90: 10, 95:5, 99: 1, or 100:0. In some aspects, the fertilizer composition comprises a ratio of inhibitor comprised in the inhibitor inclusion complex to inhibitor not comprised in an inhibitor inclusion complex of 80:20 to 100:0. In some aspects, the ratio is, is at least, is at most, or is between 80:20, 85: 15, 90: 10, 95:5, 99: 1, or 100:0.

[0017] Some aspects of the present disclosure are directed to a method of control-release of an inhibitor into a soil, a plant, water, or a combination thereof, the method comprising applying the fertilizer composition to the soil, plant, water, or combination thereof. In some aspects of the method the inhibitor is released from the inhibitor inclusion complex over a period of at least 21 days after the fertilizer composition is applied to the soil, plant, water, or combination thereof. The inhibitor may be released from the inhibitor inclusion complex over a period of at least, at most, or between, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 days.

[0018] Some aspects of the present disclosure are directed to a method of producing the fertilizer composition comprising: (a) contacting the inhibitor with at least one complexing component to generate a inhibitor inclusion complex; and (b) contacting the inhibitor inclusion complex with a nitrogen fertilizer to form an inhibitor inclusion complex embedded in the nitrogen fertilizer. In some instances, the nitrogen fertilizer is a molten nitrogen fertilizer and solidifying the molten nitrogen fertilizer forms a inhibitor inclusion complexembedded in the nitrogen fertilizer. In some aspects of the method, steps a and b are performed as a one pot synthesis and / or multi-step synthesis. In some further aspects, the method further comprises a mechanical mixing and / or cooling.

[0019] In certain aspects, the inhibitor inclusion complex is prepared by contacting a solution, such as methanol solution of NBTPT, with a solution, such as aqueous solution of y- cyclodextrin or a derivative thereof. In certain aspects, the inhibitor inclusion complex is prepared by mechanically mixing the NBTPT in its solid form with y-cyclodextrin or a derivative thereof in its solid form.

[0020] One aspect of the disclosure is directed to a method of fertilizing comprising contacting a soil, a plant, water, or a combination thereof with the disclosed fertilizer composition herein. In another aspect of the method of fertilizing, the fertilizer composition is added directly in a solid form and / or to water before contacting the soil, plant, or soil and plant.

[0021] Also disclosed are the following Aspects 1 to 44 of the present invention. Aspect 1 concerns the fertilizer composition comprising: a nitrogen fertilizer; and a cyclodextrin- inhibitor inclusion complex comprising at least one cyclodextrin and / or derivative thereof complexed with an inhibitor, wherein the inhibitor comprises at least one nitrification inhibitor and / or at least one urease inhibitor, and wherein at least one cyclodextrin-inhibitor inclusion complex is embedded in the nitrogen fertilizer.

[0022] Aspect 2 concerns the fertilizer composition of aspect 1, wherein the cyclodextrin- inhibitor inclusion complex is embedded in a solidified nitrogen fertilizer melt.

[0023] Aspect 3 concerns the fertilizer composition of any one of aspects 1 and 2, wherein the cyclodextrin-inhibitor inclusion complex is thermally stable at 140 °C or more and / or is stable at a pH range between 5 to 10.

[0024] Aspect 4 concerns the fertilizer composition of any one of aspects 1 to 3, wherein the cyclodextrin and / or derivative thereof comprises a-cyclodextrin, [3-cyclodextrin, and / or y- cyclodextrin.

[0025] Aspect 5 concerns the fertilizer composition of any one of aspects 1 to 4, wherein the nitrification inhibitor comprises 3,4-dimethylpyrazole phosphate (DMPP), thio-urea (TU),dicyandiamide (DCD), 2-chloro-6-(trichloromethyl)pyridine (nitrapyrin), 5 -ethoxy-3 - trichloromethyl- 1 ,2,4-thiadiazol (TERRAZOLE™), 2-amino-4-chloro-6-methyl -pyrimidine (AM), 2-mercapto-benzothiazole (MBT), ammonium thiosulfate (ATS), 2- sulfanimalamidothiazole (ST), or a combination thereof.

[0026] Aspect 6 concerns the fertilizer composition of any one of aspects 1 to 5, wherein the urease inhibitor comprises N-(n-butyl)- thiophosphoric triamide, N-(n-butyl)-phosphoric triamide, N-(n-propyl) thiophosphoric triamide, benzoylthiourea (BTU), hydroquinone, acetohydroxamic acid (AHA), hydroxyurea (HU), and / or phenyl phosphorodiamidate (PPDA).

[0027] Aspect 7 concerns the fertilizer composition of any one of aspects 1 to 6, comprising 90 to 99 wt. % of the nitrogen fertilizer; and / or about 0.001 to 10 wt.% of the cyclodextrin- inhibitor inclusion complex.

[0028] Aspect 8 concerns the fertilizer composition of any one of aspects 1 to 7, further comprises a wetting agent, a carrier, a fdler agent, and / or a binder.

[0029] Aspect 9 concerns the fertilizer composition of any one of aspects 1 to 8, wherein the fertilizer composition comprises a ratio of inhibitor to degradation products of the inhibitor of 80:20 to 100:0.

[0030] Aspect 10 concerns the fertilizer composition of any one of aspects 1 to 9, wherein the fertilizer composition comprises a ratio of inhibitor comprised in the cyclodextrin-inhibitor inclusion complex to inhibitor not comprised in a cyclodextrin-inhibitor inclusion complex of 80:20 to 100:0.

[0031] Aspect 11 concerns the fertilizer composition of any one of aspects 1 to 10, wherein the nitrogen fertilizer comprises urea.

[0032] Aspect 12 concerns the fertilizer composition of aspect 11, wherein the urea comprises a solidified urea melt.

[0033] Aspect 13 concerns the fertilizer composition of any one of aspects 1 to 12, wherein the fertilizer composition is homogenous.

[0034] Aspect 14 concerns the method of controlled release of an inhibitor into a soil, a plant, water, or a combination thereof, the method comprising applying the fertilizer composition of any one of aspects 1 to 13 to the soil, plant, water, or combination thereof.

[0035] Aspect 15 concerns the method of aspect 14, wherein the inhibitor is released from the cyclodextrin-inhibitor inclusion complex over a period of at least 21 days after the fertilizer composition is applied to the soil, plant, water, or combination thereof.

[0036] Aspect 16 concerns the method of producing the fertilizer composition of any one of aspects 1 to 13, the method comprising: (a) contacting the inhibitor with at least one cyclodextrin and / or derivative thereof to generate a cyclodextrin-inhibitor inclusion complex; and (b) contacting the cyclodextrin-inhibitor inclusion complex with a molten nitrogen fertilizer and solidifying the molten nitrogen fertilizer to form a cyclodextrin-inhibitor inclusion complex embedded in the nitrogen fertilizer.

[0037] Aspect 17 concerns the method of aspect 16, wherein steps a and b are performed as a one pot synthesis and / or multi-step synthesis.

[0038] Aspect 18 concerns the method of any one of aspects 16 and 17, wherein step b further comprising mechanical mixing and / or cooling.

[0039] Aspect 19 concerns the method of fertilizing, the method comprising contacting a soil, a plant, water, or a combination thereof with the fertilizer composition of any one of aspects 1 to 13.

[0040] Aspect 20 concerns the method of fertilizing of aspect 19, wherein the fertilizer composition is added directly in a solid form and / or to water before contacting the soil, plant, or soil and plant.

[0041] Aspect 21 concerns the fertilizer composition of any one of aspects 1 to 13 or the method of any one of aspects 14 to 20, wherein the cyclodextrin and / or derivative thereof is instead a polymer that is not a cyclodextrin and / or derivative thereof.

[0042] Aspect 22 concerns the fertilizer composition of aspect 21, wherein the polymer is a polysaccharide and / or a oligosaccharide.

[0043] Aspect 23 concerns the fertilizer composition of aspect 22, wherein the polymer is a cellulose, a starch, and / or a derivative thereof.

[0044] Aspect 24 concerns a fertilizer composition comprising: a solidified nitrogen fertilizer melt; said melt comprising: one or more cyclodextrin-inhibitor inclusion complexes comprising at least one cyclodextrin and / or derivative thereof complexed with an inhibitor, wherein the inhibitor comprises at least one nitrification inhibitor and / or at least one urease inhibitor, and wherein the one or more cyclodextrin-inhibitor inclusion complexes is homogeneously dispersed throughout the solidified nitrogen fertilizer melt.

[0045] Aspect 25 concerns the fertilizer composition of aspect 24, comprising: a continuous phase comprising the nitrogen fertilizer melt; and a discontinuous phase comprising the one or more cyclodextrin-inhibitor complexes dispersed throughout the continuous phase.

[0046] Aspect 26 concerns the fertilizer composition of aspect 25, wherein the one or more cyclodextrin-inhibitor complexes are evenly dispersed throughout the continuous phase.

[0047] Aspect 27 concerns the fertilizer composition any one of aspects 24 to 26, wherein the discontinuous phase comprising the nitrogen fertilizer melt has a melt temperature of 140 °C or less, preferably 130 °C to 135 °C.

[0048] Aspect 28 concerns the fertilizer composition any one of aspects 24 to 26, wherein: the nitrogen fertilizer melt comprises at least 90 wt. %, preferably at least 95 wt. %, more preferably at least 98 wt. %, or even more preferably at least 99 wt. % of the fertilizer composition; and the one or more cyclodextrin-inhibitor inclusion complexes comprises up to 10 wt. %, preferably up to 5 wt. %, more preferably up to 2 wt. %, or even more preferably up to 1 wt. % of the fertilizer composition.

[0049] Aspect 29 concerns the fertilizer composition any one of aspects 24 to 26, wherein the cyclodextrin-inhibitor inclusion complexes are thermally stable at 140 °C or more and / or are stable at a pH range between 5 to 10.

[0050] Aspect 30 concerns the fertilizer composition any one of aspects 24 to 26, wherein the cyclodextrin and / or derivative thereof comprises a-cyclodextrin, [3-cyclodextrin, and / or y- cyclodextrin, preferably y-cyclodextrin.

[0051] Aspect 31 concerns the fertilizer composition any one of aspects 24 to 26, wherein the nitrification inhibitor comprises 3,4-dimethylpyrazole phosphate (DMPP), thio-urea (TU), dicyandiamide (DCD), 2-chloro-6-(trichloromethyl)pyridine (nitrapyrin), 5-ethoxy-3- trichloromethyl- 1 ,2,4-thiadiazol (TERRAZOLE™), 2-amino-4-chloro-6-methyl-pyrimidine (AM), 2-mercapto-benzothiazole (MBT), ammonium thiosulfate (ATS), 2- sulfanimalamidothiazole (ST), or a combination thereof.

[0052] Aspect 32 concerns the fertilizer composition any one of aspects 24 to 26, wherein the urease inhibitor comprises N-(n-butyl)— thiophosphoric triamide, N-(n-butyl)-phosphoric triamide, N-(n-propyl) thiophosphoric triamide, benzoylthiourea (BTU), hydroquinone, acetohydroxamic acid (AHA), hydroxyurea (HU), and / or phenyl phosphorodiamidate (PPDA), preferably N-(n-butyl) -thiophosphoric triamide.

[0053] Aspect 33 concerns the fertilizer composition any one of aspects 24 to 26, wherein: the solidified nitrogen fertilizer melt is a urea fertilizer melt; and the inhibitor is a urease inhibitor, preferably N-(n-butyl)-thiophosphoric triamide.

[0054] Aspect 34 concerns the fertilizer composition any one of aspects 24 to 26, further comprises a wetting agent, a carrier, a filler agent, and / or a binder.

[0055] Aspect 35 concerns the fertilizer composition any one of aspects 24 to 26, wherein the fertilizer composition comprises a ratio of inhibitor to degradation products of the inhibitor of 80:20 to 100:0.

[0056] Aspect 36 concerns the fertilizer composition any one of aspects 24 to 26, wherein the fertilizer composition comprises a ratio of inhibitor comprised in the one or more cyclodextrin-inhibitor inclusion complexes to inhibitor not complexed with cyclodextrin of 80:20 to 100:0.

[0057] Aspect 37 concerns the fertilizer composition any one of aspects 24 to 26, wherein the nitrogen fertilizer melt comprises urea.

[0058] Aspect 38 concerns the fertilizer composition any one of aspects 24 to 26, wherein the fertilizer composition is homogenous.

[0059] Aspect 39 concerns a method of controlled release of an inhibitor into a soil, a plant, water, or a combination thereof, the method comprising applying the fertilizer composition of any one of aspects 24 to 38 to the soil, plant, water, or combination thereof.

[0060] Aspect 40 concerns the method of aspect 39, wherein the inhibitor is released from the one or more cyclodextrin-inhibitor inclusion complexes over a period of at least 21 days after the fertilizer composition is applied to the soil, plant, water, or combination thereof.

[0061] Aspect 41 concerns a method of producing the fertilizer composition of any one of aspects 24 to 38, the method comprising: (a) contacting the inhibitor with a one or more cyclodextrins and / or derivatives thereof to generate a one or more cyclodextrin-inhibitor inclusion complexes; and (b) contacting the one or more cyclodextrin-inhibitor inclusion complexes with a molten nitrogen fertilizer; and (c) solidifying the molten nitrogen fertilizer to form the fertilizer composition.

[0062] Aspect 42 concerns the method of aspect 41, wherein the molten nitrogen fertilizer solubilizes the one or more cyclodextrin-inhibitor inclusion complexes such that individual molecules of the complexes are dispersed throughout the molten nitrogen fertilizer.

[0063] Aspect 43 concerns the method of any one of aspects 41 to 42, wherein: steps (a) and (b) are performed as a one pot synthesis and / or multi-step synthesis and / or step (b) further comprises mixing the one or more cyclodextrin-inhibitor inclusion complexes with the molten nitrogen fertilizer.

[0064] Aspect 44 concerns a method of fertilizing, the method comprising contacting a soil, a plant, water, or a combination thereof with the fertilizer composition of any one of aspects 24 to 38, preferably wherein the fertilizer composition is added directly in a solid form and / or to water before contacting the soil, plant, or soil and plant.

[0065] The following includes definitions of various terms and phrases used throughout this specification.

[0066] The term “fertilizer” is defined as a material applied to soils or to plant tissues to supply one or more plant nutrients essential or beneficial to the growth of plants and / or stimulants or enhancers to increase or enhance plant growth. Non-limiting examples of fertilizers include materials having one or more of urea, ammonium nitrate, calciumammonium nitrate, urea calcium sulfate adduct, one or more superphosphates, binary NP fertilizers, binary NK fertilizers, binary PK fertilizers, NPK fertilizers, molybdenum, zinc, copper, boron, cobalt, and / or iron. In some aspects, fertilizers include agents that enhance plant growth and / or enhance the ability for a plant to receive the benefit of a fertilizer, such as, but not limited to biostimulants, urease inhibitors, and nitrification inhibitors.

[0067] The term “nutrient” is defined as a chemical element or substance used for the normal growth and development of a plant. Non-limiting examples of nutrients include N, P, K, Ca, Mg, S, B, Cu, Fe, Mn, Mo, Zn, Se, and Si or compounds thereof.

[0068] The term “granule” can include a solid material. A granule can have a variety of different shapes, non-limiting examples of which include a spherical, a puck, an oval, a rod, an oblong, or a random shape. The term “prill” refers to a solid globule of a substance formed by the congealing of a liquid. The term “pellet” refers to a rounded, compressed mass of fertilizer. The term “powder” refers to dry particles produced by the grinding, crushing, or disintegration of a fertilizer composition.

[0069] The terms “about” or “approximately” are defined as being close to as understood by one of ordinary skill in the art. In one non-limiting embodiment, the terms are defined to be within 10%, preferably within 5%, more preferably within 1%, and most preferably within 0.5%.

[0070] The terms “wt.%,” “vol.%,” or “mol.%” refers to a weight percentage of a component, a volume percentage of a component, or molar percentage of a component, respectively, based on the total weight, the total volume of material, or total moles, that includes the component. In a non-limiting example, 10 grams of component in 100 grams of the material is 10 wt.% of component.

[0071] The term “substantially” and its variations are defined to include ranges within 10%, within 5%, within 1%, or within 0.5%.

[0072] The terms “inhibiting” or “reducing” or “preventing” or “avoiding” or any variation of these terms, when used in the claims and / or the specification, includes any measurable decrease or complete inhibition to achieve a desired result.

[0073] The term “effective,” as that term is used in the specification and / or claims, means adequate to accomplish a desired, expected, or intended result.

[0074] The use of the words “a” or “an” when used in conjunction with any of the terms “comprising,” “including,” “containing,” or “having” in the claims, or the specification, may mean “one,” but it is also consistent with the meaning of “one or more,” “at least one,” and “one or more than one.”

[0075] The words “comprising” (and any form of comprising, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “includes” and “include”), or “containing” (and any form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.

[0076] The inhibitor inclusion complex nitrogen fertilizer composition and methods of producing such of the present invention can “comprise,” “consist essentially of,” or “consist of’ particular ingredients, components, compositions, steps, etc. disclosed throughout the specification. With respect to the transitional phase “consisting essentially of,” in one nonlimiting aspect, a basic and novel characteristic of the inhibitor inclusion complex nitrogen fertilizer composition of the present invention is the presence of a urease and / or nitrification inhibitor complexed with a complexing component(e.g., a cyclodextrin).BRIEF DESCRIPTION OF THE DRAWINGS

[0077] Advantages of the present invention may become apparent to those skilled in the art with the benefit of the following detailed description and upon reference to the accompanying drawings.

[0078] FIGS. 1(A)-(E) SEM images and EDX mapping: (A) 0.1 wt.% NBTPT coated Urea surface; (B) higher magnitude of 0.1 wt.% NBTPT coated Urea surface; (C) EDX analysis of the spherical particle; (D) Milt-mixed Urea with 0.1 wt.% NBTPT in NBTPT / CDs surface; and (E) higher magnification of 0. 1 wt% NBTPT in NBTPT / CDs surface.

[0079] FIGS. 2A and B show the thermogravimetry (TG) curves of the y-CD:NBTPT complexes listed in Table 1. FIG. 2A shows TG data over the temperature range 0 to 500 °C. FIG. 2B shows TG data over the temperature range 100 to 250 °C. In FIGS. 2A and B, foreach TG curve, the y-CD:NBTPT molar ratio of the complex for which the TG data was obtained is shown.

[0080] FIGS. 3A and B show the TG curves of the y-CD:NBTPT complexes listed in Table 1. In FIGs. 3A and B the initial bump (see FIG. 2A) due to retained moisture in y-CD is removed. FIG. 3A shows TG data over the temperature range 0 to 500 °C, and FIG. 3B shows TG data over the temperature range 80 to 250 °C. In FIGS. 3A and B, for each TG curve, the y-CD:NBTPT molar ratio of the complex for which the TG data was obtained is shown.

[0081] FIG. 4A shows TG curves of the y-CD :NBTPT: Urea complexes listed in Table 2. In FIG. 4A, for each TG curve, the y-CD:NBTPT:Urea molar ratio of the complex for which the TG data was obtained is shown. FIG. 4B shows TG-DTA curves of y-CD:NBTPT (molar ratio 1: 1), and y-CD :NBTPT: Urea (molar ratio 1: 1: 1).

[0082] FIGS. 5A-D show thermogravimetry differential thermal analysis (TG-DTA) curves of the samples B4 (Table 1), B13 (Table 3) and B14 (Table 3). FIG. 5A shows TG (left y- axis) and DTA (right y-axis) of the samples over the temperature range 0 to 500 °C; FIG. 5B shows TG of the samples over the temperature range 20 to 295 °C; FIG. 5C shows DTA of the samples over the temperature range 20 to 70 °C; and FIG. 5D shows DTA of the samples over the temperature range 200 to 300 °C.

[0083] FIGs. 6A-F show31P NMR spectra of y-CD / NBTPT complexes. FIG. 6A shows31P NMR spectra of NBTPT (not complexed); FIG. 6B shows31P NMR spectra of a complex with y-CD / NBTPT molar ratio 1:4; FIG. 6C shows31P NMR spectra of a complex with y- CD / NBTPT molar ratio 1:3; FIG. 6D shows31P NMR spectra of a complex with y-CD / NBTPT molar ratio 1:2; FIG. 6E shows31P NMR spectra of a complex with y-CD / NBTPT molar ratio 1: 1; and FIG. 6F shows31P NMR spectra of a complex with y-CD / NBTPT molar ratio 2: 1. The inset of each of FIGs. 6A-F shows degradation of NBTPT with time, determined based on the P=S peak (around 61 ppm).

[0084] FIG. 7 shows31P NMR spectrum of y-CD / NBTPT complex (1: 1 molar ratio) after 17 days.

[0085] FIGS. 8A-B show31P NMR spectra of the y-CD:NBTPT:urea and y-CD:NBTPT complexes after heat treatment. FIG. 8A shows the31P NMR of y-CD:NBTPT:urea (1: 1: 1molar ratio) at room temperature (rt), and after heat treatment at 140 °C, 160 °C, and 180 °C, and FIG. 8B shows the31P NMR of y-CD:NBTPT (1: 1 molar ratio) at rt, and after heat treatment at 140 °C, 160 °C, and 180 °C.

[0086] FIG. 9 shows Rotating-Frame Overhauser Enhancement spectrum of y-CD:NBTPT 1: 1 (molar ratio) complex. The x-axis is expanded in the bottom figure.

[0087] FIGS 10A-B show solution based Urease inhibition of NBTPT. FIG. 10A shows Half maximal inhibitory concentration (ICso) calculation of NBTPT. FIG. 10B shows urease inhibition results under various NBTPT / y-CDs molar ratio (2: 1 to 1:4) at NBTPT concertation of 0.6 pmol in all cases.

[0088] FIGS 11A-F show urease inhibition rate of heat-treated encapsulated y- CD / NBTPT(with or without Urea) complexes. FIGs. HA shows urease inhibition rate of heated encapsulated y-CD / NBTPT(with Urea) heat treated at 140 °C. FIGs. 11B shows urease inhibition rate of heated encapsulated y-CD / NBTPT(with Urea) heat treated at 160 °C. FIGs. 11C shows urease inhibition rate of heated encapsulated y-CD / NBTPT(with Urea) heat treated at 180 °C. FIGs. HD shows urease inhibition rate of heated encapsulated y- CD / NBTPT(without Urea) heat treated at 140 °C. FIGs. HE shows urease inhibition rate of heated encapsulated y-CD / NBTPT(without Urea) heat treated at 160 °C. FIGs. HF shows urease inhibition rate of heated encapsulated y-CD / NBTPT(without Urea) heat treated at 180 °C.

[0089] FIGS. 12 A-C show urease activity measurements of soil samples. FIG. 12A show urease content of various soil types. FIG. 12B shows ammonia volatilization test results before encapsulation of the samples listed in Table 8. FIG. 12C shows ammonia volatilization test results after encapsulation and heat-treatment of the samples listed in Table 8.

[0090] FIG. 13 shows ammonia production at day 15 of from the samples listed Table 8.

[0091] FIG. 14 shows Jobs plot of a y-CD:NBTPT complex.

[0092] While the invention is susceptible to various modifications and alternative forms, specific embodiments thereof are shown by way of example in the drawings. The drawings may not be to scale.DETAILED DESCRIPTION OF THE INVENTION

[0093] A fertilizer composition comprising a combination of nitrogen fertilizer and an inhibitor inclusion complex comprising at least one complexing component with an inhibitor. The inhibitor may comprise at least one nitrification inhibitor and / or at least one urease inhibitor. The inhibitor inclusion complex may be embedded in the nitrogen fertilizer. The controlled-release fertilizer composition may include an inhibitor inclusion complex comprising at least one nitrification inhibitor and / or at least one urease inhibitor component, where the inhibitor inclusion complex is encapsulated / embedded within and / or dispersed throughout a nitrogen fertilizer melt. The inhibitor inclusion complex may be encapsulated / embedded within the nitrogen fertilizer and may also be provided on the surface of the nitrogen fertilizer. The nitrogen fertilizer component may comprise urea. The inclusion complex may be broken over time and impart a controlled-release functionality for the inhibitor and may slow the loss of nitrogen in the fertilizer to nitrification or hydrolysis. Additional advantages of this fertilizer composition include the ability to provide a required complexing component size, such as cyclodextrin types such as a-cyclodextrin, [3- cyclodextrin, y-cyclodextrin and / or its derivatives thereof, according to the size of inhibitor molecule to be complexed with the complexing component. The inhibitor inclusion complex may increase the thermal stability of the inhibitor in the complex compared to inhibitor not found in an inclusion complex.

[0094] In some instances, the complexing component is a polymer. In some instances, the polymer is an oligomer. In some instances, the polymer is a polysaccharide, a oligosaccharide, and / or a derivative thereof. The polymer may be a cellulose, a starch, and / or a derivative thereof. In some instances, the polymer is a cyclodextrin. Cyclodextrins are a family of cyclic oligosaccharides containing a macrocyclic ring of glucose subunits joined by a- 1,4 glycosidic bonds. Cyclodextrins may be produced from starch by enzymatic conversion. Typical cyclodextrins may contain six to eight glucose monomer units in a ring, creating a cone shape. For example a (alpha)-cyclodcxtrin. [3 (beta), and y (gamma)- cyclodextrins contain 6, 7, and 8 glucose subunits, respectively. These cyclodextrins have toroidal shapes, with the larger and the smaller openings of the toroid exposing secondary and primary hydroxyl groups, respectively, on the outside of the ring. Because of this arrangement, the interior of the toroids is not hydrophobic, but considerably less hydrophilic than the outside of the toroid and able to host hydrophobic molecules. In contrast, the exterior is sufficiently hydrophilic to impart cyclodextrins (or their complexes) water solubility. Thesepolysaccharide and / or derivatives may be primarily obtained from natural sources such as wood pulp and other plant sources, and may constitute a renewable scaffold for the fertilizer composition disclosed herein.A. Fertilizer Compositions

[0095] The fertilizer composition can contain about 80 to 99.9999 wt. % of the nitrogen fertilizer and / or about 0.0001 to 15 wt.% of the inhibitor inclusion complex. The fertilizer composition can contain at least any one of, at most any one of, equal to any one of, or between any two of 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100 wt.% of nitrogen fertilizer. In some instances, the nitrogen fertilizer can be a solidified nitrogen fertilizer melt. In some instances, the nitrogen fertilizer can comprises an urea, urea melt, and / or NPK. In some aspects, the fertilizer composition can contain 0.0001 wt. % to 10 wt. %, or at least any one of, equal to any one of, or between any two of 0.0001 wt. %, 0.0002 wt. %, 0.0003 wt. %, 0.0004 wt. %, 0.0005 wt. %, 0.0006 wt. %, 0.0007 wt. % 0.0008 wt. %, 0.0009 wt. %, 0.001 wt. %, 0.002 wt. %, 0.003 wt. %, 0.004 wt. %, 0.005 wt. %, 0.006 wt. %, 0.007 wt. % 0.008 wt. %, 0.009 wt. %, 0.01 wt. %, 0.02 wt. %, 0.03 wt. %, 0.04 wt. %, 0.05 wt. %, 0.06 wt. %, 0.07 wt. % 0.08 wt. %, 0.09 wt. %, 0.1 wt. %, 0.2 wt. %, 0.3 wt. %, 0.4 wt. %, 0.5 wt. %, 0.6 wt. %, 0.7 wt. % 0.8 wt. %, 0.9 wt. %, 1 wt. %, 1.2 wt. %, 1.4 wt. %, 1.6 wt. %, 1.8 wt. %, 2 wt. %, 2.2 wt. %, 2.4 wt. %, 2.6 wt. %, 2.8 wt. %, 3 wt. %, 3.2 wt. %, 3.4 wt. %, 3.6 wt. %, 3.8 wt. %, 4 % 4.2 wt. %, 4.4 wt. %, 4.6 wt. %, 4.8 wt. %, 5 wt. %, 5.2 wt. %, 5.4 wt. %, 5.6 wt. %, 5.8 wt. %, 6 %, 6.2 wt. %, 6.5 wt. %, 7 wt. %, 7.5 wt. %, 8 wt. %, 8.5 wt. %, 9 wt. %, 9.5 wt. %, 10 wt.% of inhibitor inclusion complex based on the total weight of the fertilizer composition. In some instances, the fertilizer composition further comprises a wetting agent, a carrier, a filler agent, and / or a binder.

[0096] In some instances, the complexing component comprises a cyclodextrin. In some instances, the inhibitor inclusion complex comprises a-cyclodextrin, [3-cyclodextrin, y- cyclodextrin and / or derivatives thereof. In some instances, the inhibitor inclusion complex comprises at least one nitrification inhibitor and / or at least one urease inhibitor. In some instances, the nitrification inhibitor comprises 3,4-dimethylpyrazole phosphate (DMPP), thiourea (TU), dicyandiamide (DCD), 2-chloro-6-(trichloromethyl)pyridine (nitrapyrin), 5- ethoxy-3 -trichloromethyl- 1, 2, 4-thiadiazol (TERRAZOLE™), 2-amino-4-chloro-6-methyl- pyrimidine (AM), 2-mercapto-benzothiazole (MBT), ammonium thiosulfate (ATS), 2- sulfanimalamidothiazole (ST), or a combination thereof. In some instances, the ureaseinhibitor comprises N-(n-butyl)-thiophosphoric triamide, N-(n-butyl)-phosphoric triamide, N- (n-propyl) thiophosphoric triamide, benzoylthiourea (BTU), hydroquinone, acetohydroxamic acid (AHA), hydroxyurea (HU), and / or phenyl phosphorodiamidate (PPDA). In some instances, the inhibitor inclusion complex may include both a nitrification inhibitor and a urease inhibitor, such as N-(n-butyl)-phosphoric triamide and dicyandiamide, such asNITRIFICIN™.

[0097] In some instances, the fertilizer composition comprises a ratio of inhibitor to degradation products of the inhibitor of 80:20 to 100:0 or at least any one of, equal to any one of, or between any two of 81: 19, 82: 18, 83: 17, 84: 16, 85: 15, 86: 14, 87: 13, 88: 12, 89: 11, 90: 10, 91:9, 92:8, 93:7, 94:6, 95:5, 96:4, 97:3, 98:2, 99: 1, and 100:0.

[0098] In some instances, the fertilizer composition comprises a ratio of inhibitor comprised in the inhibitor inclusion complex to inhibitor not comprised in an inhibitor inclusion complex of 80:20 to 100:0 or at least any one of, equal to any one of, or between any two of 81: 19, 82: 18, 83: 17, 84: 16, 85: 15, 86: 14, 87: 13, 88: 12, 89: 11, 90: 10, 91:9, 92:8, 93:7, 94:6, 95:5, 96:4, 97:3, 98:2, 99: 1, and 100:0.B. Method of Making the Fertilizer Composition

[0099] A method of producing the fertilizer composition comprising inhibitor inclusion complex encapsulated within nitrogen fertilizer is disclosed. In some aspects, the method comprising the following steps: (a) contacting the inhibitor with at least one complexing component to generate an inhibitor inclusion complex; and (b) contacting the inhibitor inclusion complex with a nitrogen fertilizer to form an inhibitor inclusion complex embedded in the nitrogen fertilizer.

[0100] In step a) above, a suitable complexing component material is first identified. In some aspects, the complexing component can be a a-cyclodextrin and / or its derivatives thereof. In some aspects, the cyclodextrin material can be a [3-cyclodextrin and / or its derivatives thereof. In some aspects, the cyclodextrin material can be a y-cyclodextrin and / or its derivatives thereof. In some aspects, any of the above mentioned cyclodextrin derivatives may replace with other complexing components, such as polymers, such as other polysaccharides, oligosaccharides, and / or derivatives thereof, such as a cellulose, a starch, and / or a derivative thereof. In some aspects, the inhibitor may be a urease inhibitor or a nitrification inhibitor, or a combination thereof. In one aspect, a urease inhibitor and anitrification inhibitor are included. In one aspect, the inhibitor may be a urease inhibitor. Suitable urease inhibitors include, but are not limited to, N-(n-butyl)-thiophosphoric triamide (NBTPT), N-(n-butyl)-phosphoric triamide (NBPT), N-(n-propyl) thiophosphoric triamide (NPPT), benzoylthiourea (BTU), hydroquinone, acetohydroxamic acid (AHA), hydroxyurea(HU), and / or phenyl phosphorodiamidate (PPDA). In one aspect, the inhibitor may comprise NBTPT, NBPT, NPPT, and / or PPDA, or a combination thereof. In another aspect, the inhibitor may be a nitrification inhibitor. Suitable nitrification inhibitors include, but are not limited to, 3,4-dimethylpyrazole phosphate (DMPP), dicyandiamide (DCD), thiourea (TU), 2-chloro-6-(trichloromethyl)-pyridine (Nitrapyrin), 5 -ethoxy-3 -trichloromethyl- 1 ,2,4- thiadiazol, which is sold under the tradename TERRAZOLE®, by OHP Inc., USA, 2-amino 4-chloro 6-methyl pyrimidine (AM), 2-mercaptobenzothiazole (MBT), or 2- sulfanilamidothiazole (ST), and any combination thereof. In one aspect, a nitrification inhibitor may comprise DMPP, DCD, TU, nitrapyrin, 5 -ethoxy-3 -trichloromethyl- 1,2,4- thiadiazol, AM, MBT, or ST, or a combination thereof. In one aspect, the fertilizer composition may comprise NBTPT, DMPP, TU, DCD, PPDA, nitrapyrin, 5 -ethoxy-3 - trichloromethyl- 1, 2, 4-thiadiazol, AM, MBT, or ST, or a combination thereof. Combining an urease inhibitor and / or a nitrification inhibitor with the above-mentioned suitable complexing component in a mixing vessel, and mixing under appropriate temperatures and pressures generates an inhibitor inclusion complex. In some instances, the inhibitor inclusion complex is formed in 1-2 hours. In some aspects, the mixture may be heated at a temperature ranging from about 40 °C to about 200 °C. In certain aspects, the formed inhibitor inclusion complex is thermally stable at 130 °C or more, such as 140 °C or more, or at least any one of, equal to any one of, or between any two of 130 °C, 145 °C, 150 °C, 155 °C, 160 °C, 165 °C, 170 °C, 175 °C, 180 °C, 185 °C, 190 °C, 195 °C, 150, 160, 170, 180, 190, 200 °C, 210 °C, 220 °C, 230 °C, 240 °C, 250 °C, 260 °C, 270 °C, 280 °C, 290 °C, and 300 °C. In certain aspects, the formed inhibitor inclusion complex is stable at a pH range between 4 to 11, such as 5 to 10, or at least any one of, equal to any one of, or between any two of 4, 5, 6, 7, 8, 9, 10 and 11.

[0101] In step b) above, the inhibitor inclusion complex in step-a, may be contacted with a solid or molten nitrogen fertilizer material or a solution of nitrogen fertilizer and mixing at a suitable-pressure and temperature (e.g., 65 °C, 40 MPa) to form inhibitor inclusion complex embedded in the nitrogen fertilizer. The inhibitor inclusion complex may be embedded in a uniform and stable nitrogen fertilizer. In some aspects, the inhibitor inclusion complex and the nitrogen fertilizer (e.g., urea) can be contacted at temperatures upto 250 °C, or at least any one of, equal to any one of, or between any two of 20 °C, 30 °C, 40°C, 45 °C, 50 °C, 55 °C, 60 °C, 65 °C, 70 °C, 75 °C, 80 °C, 85 °C, 90 °C, 95 °C, 100 °C, 105°C, 110 °C, 115 °C, 120 °C, 125 °C, 130 °C, 135 °C, 140 °C, 145 °C, 150 °C, 155 °C, 160°C, 165 °C, 170 °C, 175 °C, 180 °C, 185 °C, 190 °C, 195 °C, 200 °C, 205 °C, 210 °C, 215°C, 220 °C, 225 °C, 230 °C, 235 °C, 240 °C, 245 °C, and 250 °C. In some aspects, steps a and b in the method are performed as a one pot synthesis and / or multi-step synthesis. Upon solidifying the molten nitrogen fertilizer with subsequent mechanical mixing and / or cooling results the fertilizer composition comprising inhibitor inclusion complex embedded in the nitrogen fertilizer. In some aspects the source of the nitrogen fertilizer is urea. In some aspects the source of the nitrogen fertilizer is NPK.

[0102] The steps depicted above provide a fertilizer that includes inhibitors (e.g., nitrification inhibitors and / or urease inhibitors) that may be non-covalently bonded to or spatially enclosed in a complexing component host material. These inhibitor inclusion complex may be physically embedded in the nitrogen fertilizer and may be found on the surface of the nitrogen fertilizer material. In some aspects, water may then be removed from this mixture. In some instances, the steps above provide an inhibitor inclusion complex embedded / encapsulated in a nitrogen fertilizer, such as a solidified nitrogen fertilizer melt. Different techniques can be employed before, during, or after drying to provide the solid fertilizer composition in powder, prill, granule, or pellet form.

[0103] The fertilizer composition produced can, in some instances, contain low amounts of moisture. The free-moisture content of the fertilizer composition may be less than 0.6 wt.%, less than 0.5 wt.% water or 0.25 wt.% to less than 0.6 wt.% water. In some instances, the free moisture content is, is less than, is more than, or is between 0.5, 0.4, 0.3, 0.2, 0.1, or 0 wt.%, or any range thereof.

[0104] The fertilizer composition may be produced in powder, prill, granule, or pellet form. In certain non-limiting aspects, the powder may comprise particles having an average particle size that is, is less than, is more than, or is between 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, or 900 micrometers or any range thereof. In some embodiments, the particles may be elongated particles or may be substantially spherical particles or other shapes, or combinations of such shapes. Non-limiting examples of shapes include a sphere, a puck, an oval, a rod, an oblong, or a random shape.

[0105] The fertilizer composition may in some instances contain a coating on the surface of the fertilizer composition. In some instances, the coating may include nutrients for a plant, additional inhibitors, such as urea hydrolysis and / or nitrification inhibitors, agents to slow or increase the rate of degradation of the granule and / or fertilizers, agents to repel moisture and / or provide a hydrophobic layer, agents that decrease or increase the reactivity of the granule and / or fertilizers, agents that provide additional benefits to plants, agents that increase the stability and / or crush strength of the granule and / or fertilizers, pH buffering agents, drying agents, biostimulants, microorganisms, etc. or any combination thereof. The coating may be a commercially available coating, an oil, a fertilizer, a micronutrient, talc, a seaweed and / or seaweed extract, a bacteria, a wax, etc. In some instances, the coating may contain surfactants. In some instances, the coating contains a wax, surfactants, and / or an amine -based compound.

[0106] In some aspects, the fertilizer composition contains a coating on the fertilizer composition, such as a core comprising inhibitor inclusion complex encapsulated within a nitrogen fertilizer, and a shell coating provided around the core. In some aspects, the fertilizer composition contains the inhibitor inclusion complex within a matrix of the nitrogen fertilizer, such as an extruded fertilizer granule.

[0107] In some aspects, the fertilizer composition contains the nitrogen fertilizer as a matrix containing inhibitor inclusion complex, complexing components, and / or inhibitors and optionally other ingredients such as additional nutrient(s), inhibitor(s), alkali material, acidic material, biostimulant(s), microorganism(s), etc. In some aspects, the fertilizer composition contains a core with a coating, the inhibitor inclusion complex and nitrogen fertilizer encapsulated within the coating. The core may contain nutrients for a plant, inhibitors of urea hydrolysis and / or nitrification, agents to slow or increase the rate of degradation of the granule, agents to repel moisture and / or provide a hydrophobic layer, agents that decrease or increase the reactivity of the granule, agents that provide additional benefits to plants, agents that increase the stability and / or crush strength of the granule, pH buffering agents, drying agents, microorganisms, etc. or any combination thereof.

[0108] The fertilizer composition disclosed herein may also be included in a blended or compounded fertilizer composition comprising other fertilizers, such as other fertilizer granules. Additional fertilizers may be chosen based on the particular needs of certain types of soil, climate, or other growing conditions to maximize the efficacy of the fertilizercomposition in enhancing plant growth and crop yield. The other fertilizer granules may be granules of urea, single super phosphate (SSP), triple super phosphate (TSP), ammonium sulfate, monoammonium phosphate (MAP), diammonium phosphate (DAP), muriate of potash (MOP), and / or sulfate of potash (SOP), and the like.C. Method of Using the Fertilizer Composition

[0109] The fertilizer composition disclosed herein can be used in methods of increasing the amounts of nitrogen, and optionally phosphorus and / or potassium in soil and of enhancing plant growth. Such methods may include applying to the soil an effective amount of a composition comprising the fertilizer composition disclosed herein. The method may include increasing the growth and yield of crops, trees, ornamentals, etc. such as, for example, palm, coconut, rice, wheat, com, barley, oats, and soybeans. The method may include applying the fertilizer composition disclosed herein to at least one of a soil, an organism, a liquid carrier, a liquid solvent, etc.

[0110] Non-limiting examples of plants that may benefit from the fertilizer disclosed herein include vines, trees, shrubs, stalked plants, fems, etc. The plants may include orchard crops, vines, ornamental plants, food crops, timber, and harvested plants. The plants may include Gymnosperms, Angiosperms, and / or Pteridophytes. The Gymnosperms may include plants from the Araucariaceae, Cupressaceae, Pinaceae, Podocarpaceae, Sciadopitaceae, Taxaceae, Cycadaceae, and Ginkgoaceae families. The Angiosperms may include plants from the Aceraceae, Agavaceae, Anacardiaceae, Annonaceae, Apocynaceae, Aquifoliaceae,Araliaceae, Arecaceae, Asphodelaceae, Asteraceae, Berberidaceae, Betulaceae, Bignoniaceae, Bombacaceae, Boraginaceae, Burseraceae, Buxaceae, Canellaceae, Cannabaceae, Capparidaceae, Caprifoliaceae, Caricaceae, Casuarinaceae, Celastraceae, Cercidiphyllaceae, Chrysobalanaceae, Clusiaceae, Combretaceae, Comaceae, Cyrillaceae, Davidsoniaceae, Ebenaceae, Elaeagnaceae, Ericaceae, Euphorbiaceae, Fabaceae, Fagaceae, Grossulariaceae, Hamamelidaceae, Hippocastanaceae, Illiciaceae, Juglandaceae, Lauraceae, Lecythidaceae, Lythraceae, Magnoliaceae, Malpighiaceae, Malvaceae, Melastomataceae, Meliaceae, Moraceae, Moringaceae, Muntingiaceae, Myoporaceae, Myricaceae,Myrsinaceae, Myrtaceae, Nothofagaceae, Nyctaginaceae, Nyssaceae, Olacaceae, Oleaceae, Oxalidaceae, Pandanaceae, Papaveraceae, Phyllanthaceae, Pittosporaceae, Platanaceae, Poaceae, Polygonaceae, Proteaceae, Punicaceae, Rhamnaceae, Rhizophoraceae, Rosaceae, Rubiaceae, Rutaceae, Salicaceae, Sapindaceae, Sapotaceae, Simaroubaceae, Solanaceae,Staphyleaceae, Sterculiaceae, Strelitziaceae, Styracaceae, Surianaceae, Symplocaceae, Tamaricaceae, Theaceae, Theophrastaceae, Thymelaeaceae, Tiliaceae, Ulmaceae, Verbenaceae, and / or Vitaceae family.

[0111] The effectiveness of compositions comprising the fertilizer composition disclosed herein may be ascertained by measuring the amount of nitrogen, phosphorus, potassium, or nitrogen, phosphorus, and potassium in the soil at various times after applying the fertilizer composition to the soil. It is understood that different soils have different characteristics, which may affect the stability of the nitrogen in the soil. The effectiveness of a fertilizer composition may also be directly compared to other fertilizer compositions by doing a side-by-side comparison in the same soil under the same conditions.

[0112] In one aspect, the fertilizer composition disclosed herein may have a density that is greater than water. This may allow the granules and / or fertilizers to sink in water rather than float. This may be especially beneficial in instances where application is intended to a crop that is at least partially or fully submerged in water. A non-limiting example of such a crop is rice, as the ground in a rice paddy is typically submerged in water. Thus, application of fertilizer composition to such crops may be performed such that the granules and / or fertilizer are homogenously distributed on the ground that is submerged under water. By comparison, granules and / or fertilizers that have a density that is less than water would have a tendency to remain in or on the water surface, which could result in washing away and / or coalescence of the granules and / or fertilizers, either of which would not achieve homogenous distribution of the granules and / or fertilizers to the ground that is submerged under water.EXAMPLES

[0113] The present invention will be described in greater detail by way of specific examples. The following examples are offered for illustrative purposes only, and are not intended to limit the invention in any manner. Those of skill in the art will readily recognize a variety of noncritical parameters which can be changed or modified to yield essentially the same results.Example 1Methods of Making Fertilizer Particles and Analyzing their PropertiesA. Materials

[0114] Technical grade urea was obtained from SABIC, Riyadh, Saudi Arabia. N-(n- butyl)-thiophosphoric triamide (NBTPT) powder was be obtained from SAMICH (HK) Ltd., Hangzhou, China, or Tokyo Kasei Kogyo Co. DCD powder was obtained from SIGMA ALDRICH / ALZCHEM, Germany. Cyclodextrin can be obtained from Fujifilm Wako Pure Chemical Industries, Ltd.

[0115] FIGs. 1A and IB show that NBTPT coated on urea pellets appears as small spheres with particle sizes ranging from 50 to 150 pm, predominantly on the surface. In contrast, the melt-mixed samples exhibit a uniform dispersion of NBTPT throughout the urea, with no NBTPT particles visible on the surface (Figures ID and IE). EDX analysis indicates that the spherical particles have a high phosphorus (P) and sulfur (S) content. In contrast, the urea surface has no detectable P and S, suggesting that the spherical particles are NBTPT, which is not well-mixed with the urea. The corresponding amounts of NBTPT in methanol were spray-coated onto the urea pellets and air-dried. The melted urea with NBTPT / CyDs was prepared by adding the corresponding amounts of NBTPT / CyDs to the melted urea at 140°C.B. Procedure for Preparing a Cyclodextrin-Inhibitor Inclusion Complex and Characterization of the Same

[0116] As explained below, cyclodextrin-inhibitor inclusion complexes were prepared by mixing NBTPT and / or DCD with cyclodextrin mechanically and / or with heat and / or solvents to form the cyclodextrin-inhibitor inclusion complex. The amount of cyclodextrin and NBTPT and / or DCD in total can be between 0.0001 to 15 wt.% relative to the weight of the complete fertilizer composition (including the nitrogen fertilizer). In some instances, no other components are used. In some instances, additional components are used in varying amounts. In some instances, other complexing components are used in addition to the cyclodextrin or as a replacement for the cyclodextrin to complex with the inhibitors. In some instances, other nitrification and / or urease inhibitors are used in addition to the NBTPT and / or DCD or as a replacement for the NBTPT and / or DCD.Bl. Thermogravimetry Differential Thermal Analysis (TG / DTA) analysis of NBTPT gamma-cyclodextrin (y-CD) complex

[0117] Solvent based methods for preparation of samples containing y-CD and NBTPT : Aqueous y-CD solutions were mixed with NBTPT solutions in methanol to prepare the samples listed in Table 1. The steps involved in preparing the samples (B1-B7) include:

[0118] Preparing stock solutions: NBTPT was dissolved in methanol ([Sol. F]) and y- CD was dissolved in distilled water ([Sol. Clx]). Dilutions of [Sol. Clx] were created ([Sol. C2x] to [Sol. C8x]).

[0119] Mixing solutions: Solutions were mixed at different volumes to achieve desired y-CD:NBTPT molar ratios.

[0120] Sample preparation: Each mixed solution was divided into sample bottles (1 mL each, containing 10 mg NBTPT).

[0121] Freeze-drying: Samples were pre-frozen and freeze-dried, then stored under dry conditions.Table 1. y-CD:NBTPT samples*DW = Distilled Water

[0122] Solvent based methods for preparation of samples containing y-CD,NBTPT and urea: Samples containing y-CD, NBTPT and urea were prepared, listed in Table 2. The steps involved in preparing the samples (B8 -Bl 1) include:

[0123] Sample Preparation (liquid method): Samples were prepared with varying molar ratios of y-CD, NBTPT, and urea, by mixing solutions of y-CD, NBTPT, and urea.

[0124] Freeze-drying: The prepared samples were freeze-dried.Table 2. y-CD:NBTPT:Urea containing samples

[0125] Mechanical mixing based methods for preparation of the samples.Mechanical mixing based preparation of y-CD:NBTPT samples includes mechanically mixing y-CD and NBTPT using a mortar and pestle. Mechanical mixing based preparation of y-CD:NBTPT:Urea samples includes mechanically mixing y-CD, NBTPT and urea using a mortar and pestle. Table 3 lists samples prepares based on mechanical mixing.Table 3. Samples prepared by mechanical mixing

[0126] Results: The TGA study investigated the effects of y-CD and urea on the thermal stability of NBTPT. Weight loss curves (TG curves) were analyzed for samples with and without urea. The TG data shows that the presence of y-CD increased the temperature at which NBTPT experienced its second major weight loss event, indicating improved thermal stability, see FIGs. 2A and B. The presence of urea made it a bit more challenging to precisely quantify the stabilization effect. Additionally, the method of preparing the y- CD:NBTPT complex had a significant impact on its thermal properties. Samples prepared by a solvent-based method differed in their melting points and decomposition behavior compared to those prepared by mechanical mixing.

[0127] FIGS. 2A and B show the TG curves of the samples Bl- B7. The temperature is plotted on the x-axis and the TG is plotted on the y-axis. FIG. 2A shows the data over the temperature range 0 to 500 °C, and FIG. 2B shows the data over the temperature range 100 to 250 °C. The initial weight loss of complex with y-CD was due to water encapsulated in y- CD. FIGs. 3A and B show the TG curves of the samples B1-B7 omitting the initial retained moisture. FIG. 3A shows the data over the temperature range 0 to 500 °C, and FIG. 3Bshows the data over the temperature range 80 to 250 °C. The raw data indicated that y-CD had 10.1% moisture. Based on FIGs. 3A and B, the second weight loss for y-CD:NBTPT complexes occurred at temperatures higher than NBTPT only. Therefore, the thermal stabilization of NBTPT is achieved using y-CD. The ratios shown in FIGS. 2A-B and 3A-B are y-CD:NBTPT molar ratio.

[0128] FIG. 4A shows the TG curves of the samples B8- B12. The samples were not heated prior to the experiment. The ratios shown in FIG. 4A is y-CD :NBTPT: Urea molar ratio. FIG. 4B shows TG-DTA curves of sample B4 (y-CD:NBTPT molar ratio 1: 1) and B10 (y-CD:NBTPT:Urea molar ratio 1: 1: 1). Two curves were slightly different in first weight loss part. It was thought that the phenomena were derived from the amount of water included y- CD.

[0129] FIGs. 5A-D shows TG and DTA curves of the samples B4, B13 and B14. As shown in Table 1, B4 was prepared using solvent based methods and has y-CD:NBTPT molar ratio 1: 1. As shown in Table 3, B13 was prepared using mechanical mixing based preparation methods and has y-CD:NBTPT molar ratio of 1: 1; B14 was prepared using mechanical mixing based preparation methods and has y-CD :NBTPT: Urea molar ratio of 1: 1: 1. FIG. 5A shows TG (left y-axis) and DTA (right y-axis) of the samples over the temperature range 0 to 500 °C; FIG. 5B shows TG of the samples over the temperature range 20 to 295 °C; FIG. 5C shows DTA of the samples over the temperature range 20 to 70 °C; and FIG. 5D shows DTA of the samples over the temperature range 200 to 300 °C. FIGs. 5A-D show that the melting point (DTA) of NBTPT was around 60° C, and the decomposition temperature was different when mechanical mixing based preparation method was used. This indicated that mechanically mixed reagents may differ from lyophilized reagents.B2. Stability study of the NBTPT y-cyclodextrin (y-CD) complex in acidic medium

[0130] Stability of the NBTPT y-cyclodextrin (y-CD) complexes in acidic medium were studied by determining degradation of NBTPT in an acidic environment using31P NMR spectroscopy.

[0131] Sample preparation: Freeze-dried samples (prepared using the methods described in section Bl) containing y-CD and NBTPT (approximately 10 mg) was dissolved in water (250 mL) using sonication.

[0132] Acidic Environment: An acidic buffer solution (CHsCOOH / CFLCOONa, 500 mM, pH 5.0) is added to the sample solution (250 mL). pH= 5 is the extreme condition for any plant.

[0133] NMR Analysis: An H3PO4 solution in D2O (placed in a capillary) is used as the reference for the31P NMR experiment.

[0134] The degradation of NBTPT is specifically monitored by observing the signal at 61 ppm on the31P NMR spectrum.

[0135] 31P NMR spectra of y-CD / NBTPT complexes are shown in FIGs. 6A-F. FIG.6A shows31P NMR spectra of NBTPT (not complexed); FIG. 6B shows31P NMR spectra of a complex with y-CD / NBTPT molar ratio 1:4; FIG. 6C shows31P NMR spectra of a complex with y-CD / NBTPT molar ratio 1:3; FIG. 6D shows31P NMR spectra of a complex with y-CD / NBTPT molar ratio 1:2; FIG. 6E shows31P NMR spectra of a complex with y- CD / NBTPT molar ratio 1: 1; and FIG. 6F shows31P NMR spectra of a complex with y-CD / NBTPT molar ratio 2: 1. The inset in each figure shows degradation of NBTPT with time, determined based on the P=S peak (around 61 ppm)

[0136] FIGs. 6A-F, show that the y-CD / NBTPT complexes were not stable under acidic conditions in liquid, the P=S peak of NBTPT decreased in 5 hours.

[0137] 31P NMR data of y-CD / NBTPT 1:1 complex after 17 days:31P NMR of y-CD:NBTPT=1: 1 (molar ratio) complex in acetic acid buffer after 17 days was measured. As can be seen from FIG. 7, the P=S peak (around 60 ppm) completely disappeared, a P=O peak (around 52 ppm) was not detected, and only the 42 ppm peak remained. This result was observed in other complexes.B3. Thermal stability study of the NBTPT y-cyclodextrin (y-CD) complex

[0138] Preparation of samples through heat treatment. Samples containing y-CD, and NBTPT, and samples containing y-CD, NBTPT and urea were prepared for heating experiments. The freeze-dried samples, (prepared using the methods described in section Bl),were heated at 140 °C, 160 °C, and 180 °C for 30 minutes each. The samples were studied using31P-NMR and1H-NMR.

[0139] FIG. 8A shows the31P NMR of y-CD:NBTPT:urea (1: 1: 1 molar ratio) at room temperature (rt), and after heat treatment at 140 °C, 160 °C, and 180 °C, and FIG. 8B shows the31P NMR of y-CD:NBTPT (1: 1 molar ratio) at rt and after heat treatment at 140 °C, 160 °C, and 180 °C. As seen in FIGs. 8A and B, the P=S peak remained after heating. Another peak appeared in 'H-NMR after heating (FIG. 9). The amount of NBTPT remaining in the gCD:NBTPT=l: l and 1:2 samples were calculated, and a decrease of NBTPT was observed as the heating temperature increased regardless of whether urea was present or not. Table 4 shows the integral of P=S from FIG. 8A, and Table 5 shows the integral of P=S from FIG. 8B.Table 4:31P NMR of y-CD:NBTPT:urea (1: 1: 1 molar ratio)Table 5:31P NMR of y-CD:NBTPT (1: 1 molar ratio)B4. Determination of the association constant between y-CD and NBTPT

[0140] Benesi-Hildebrand method was used to determine the association constant (Ka) which describes the strength of the interaction between NBTPT and y-CD. Methanolbased NBTPT solution (5 mM) and aqueous y-CD solution (125 mM) were prepared.Varying amounts of these solutions were mixed with water to achieve a final volume of 500pL, maintaining a constant NBTPT concentration (ImM) and a range of y-CD concentrations (50-100mM). These mixtures were then freeze-dried, and the resulting solids were dissolved in D2O for 'H NMR analysis. The variation of chemical shift (A5i [CD]) for each proton (i) of NBTPT was calculated at each y-CD concentration using the following equations:1 intercept = —:— slopeA3’ Cpx

[0141] Plotting 1 / A5i [y-CD] against l / [y-CD] yielded linear relationships. The association constant (Ka) and the chemical shift changes of NBTPT in the pure complex (A5i Cpx) were determined from the equations for the intercept and slope of these lines.

[0142] Based on NMR measurement of NBTPT / y-CD =1: 1 (molar ratio) complex, the shift value (Dd) of butyl-chain of NBTPT was calculated. The results were listed in Table 6 and the calculated Ka was 5.17±6.28.Table 6: Association constant between y-CD and NBTPT.B5. Rotating-Frame Overhauser Enhancement Spectroscopy (ROESY) measurement of y-CD:NBTPT 1:1 (molar ratio) complex

[0143] NMR samples of y-CD:NBTPT 1: 1 (molar ratio) complex (lOmg) were prepared and ROESY was measured on the 1: 1 complex in D2O. As can be seen from the ROSEY spectrum (FIG. 9) the butyl-chain on NBTPT might interact with H3 and H5 of y- CD and the complex is formed.B6. Evaluation of the Urease Inhibitory Activity

[0144] Preparation of NBTPT Solutions: Stock Solution: NBTPT was dissolved in methanol (20 mg / mL or 120 mM concentration). Working Solutions: The stock solution was diluted with Tris-HCl buffer (pH=8.0) to create solutions with varying NBTPT concentrations (0-600 pM).

[0145] Preparation of Freeze-dried Samples. Dissolving: Freeze-dried samples were dissolved in methanol (20 mg / mL or 120 mM NBTPT). Centrifugation: The solution was centrifuged to remove any insoluble material. Dilution: The supernatant was diluted with Tris-HCl buffer (pH=8.0) to a 6 mM NBTPT concentration.

[0146] Urease Inhibition Assay (Jackbean Urease): Urease solution (derived from Jackbean, Fujifilm Wako Pure Chemicals) was prepared to 2 pg / mL in buffer (50 mM Tris- HCl pH 8.0). After mixing 100 pl of the prepared urease solution with 100 pl of the sample solution prepared, the mixture was incubated at 37°C for 30 minutes.

[0147] Ammonia Detection: Glutamate dehydrogenase enzyme system was used to quantify ammonia produced and the chemicals and concentrations used in the essay are shown in Table 7. Absorbance change at 340 nm (NADPH decrease) was measured over 20 minutes, providing an indication of urease activity. The peak intensity decreases while NADPH deprotonates to NADP+by reacting with ammonia.

[0148] Urease assay: Urea + H2O = CO2 + NH3 (Urease). KGA + NH3 + NADPH = L-Glutamate +NADP+ + H2O (Glutamate dehydrogenase). The decrease in absorbance at 340 nm, due to the oxidation of NADPH, is proportional to the ammonia concentration. L- Glutamate dehydrogenase reacts specifically with ammonia.Table 7: Composition of Urea inhibition assay

[0149] Calculations: Enzyme activity is defined as 1 U = 1 mmol urea degraded per minute. Measurements were done in triplicate for accuracy.

[0150] Results: Encapsulation by y-CD was expected to reduce the urease inhibitory activity of NBTPT. The aim of this study was to evaluate the urease inhibitory activity of the prepared encapsulated y-CD / NBTPTs by enzymatic methods and to select samples that maintained urease inhibition activity. The urease inhibitory activity of heat-treated encapsulated y-CD / NBTPTs was also evaluated. As shown in FIG. 10A, the half maximal inhibitory concentration (ICso) of NBTPT is around 0.65 pM. The 0.60 pM was used during the solution based inhibition study. Regardless the NBTPT / y-CDs, the final concentration of NBTPT was always kept as 0.60 pM to have a fair comparison.

[0151] The urease inhibition efficiency of pure NBTPT was decreased from 61.5% to 40% after freeze-drying, indication relatively low stability under room temperature in the water (FIG. 10A). In contrast, samples encapsulated with y-CD (2: 1, 1: 1, 1:2, 1:3, 1:4 (NBTPT / y-CDs molar ratio)) did not show any decrease in urease inhibition, and stabilization of NBTPT by inclusion was observed (FIG. 10B). Interestingly, the pure y-CD showed around 30% of inhibition efficiency, suggesting non-competitive inhibition nature (FIG. 10B)

[0152] The urease inhibition efficiency of heat-treated encapsulated y- CD / NBTPT(with or without Urea) under various temperature are shown in FIGs. 11A-F. FIGs. 11 A-C show urease inhibition rate of heated encapsulated y-CD / NBTPT(with Urea). No reduction in NBTPT activity was observed when y-CD / NBTPT was 2: 1, 1: 1 or 1:2 at 140°C, 160°C or 180°C treatment respectively; no activity was observed at 1:4 at any temperature range. FIGs. 11 D-F show The urease inhibition rate of heated encapsulated y- CD / NBTPT without urea. Inclusion with y-CD increased the activity of NBPT in each of the 140°C, 160°C and 180°C treatments. The y-axis in each of FIGs. 11A-F shows urease inhibition (%). The head-treatment was performed under N2 and the NBTPT concentrationwas 6 pM during the inhibition tests. The inhibition efficiency of all the encapsulated samples were enhanced after heat-treatment along with the Urea. The enhancement is especially significant in the case of heat-treatment at 140 °C.

[0153] Evaluation of Soil Urease Activity: Purpose: To choose soils with appropriate urease activity for further testing. Procedure: Urea solution (80 mM) is mixed with soil samples. Incubation at 37 °C for 1 hour. Ammonia produced is measured (enzymatically). Enzyme activity is calculated as above.

[0154] Urea Hydrolysis Inhibition Test in Soils: Purpose: Evaluate urease inhibition by: NBTPT alone; Encapsulated NBTPT + y-CD; Mechanically mixed NBTPT + y-CD. Effect of heating on these samples.

[0155] Table 8 lists the samples used for Soil Urease Activity, outlines specific conditions for soil tests, and details sample preparation methods for the different inhibitor formulations.

[0156] Overall Aim: These experiments were designed to assess how well NBTPT, alone or in combination with y-CD, can inhibit urease activity in real soil environments. This helps understand the potential of NBTPT-based formulations for improving fertilizer efficiency.

[0157] Soil Preparation:

[0158] Adding Urea: Soil was collected from vegetable fields. Urea granules (2 g) were mixed into 500 g of soil. This gives a final urea concentration of 0.4% in the soil, simulating fertilizer application.

[0159] Adding Samples: Table 9 outlines the different ways samples (NBTPT formulations) were added to the soil. Samples were dissolved in distilled water (200 pL). This solution was added to 10 g portions of the soil. Control samples (no NBTPT) also receive 200 pL of distilled water to maintain consistent moisture.

[0160] Soil Ammonia Volatilization Test

[0161] Setup: 10 g of prepared soil was placed into a glass vial and sealed tightly. Vials were kept at 30°C in a controlled environment.

[0162] Ammonia Measurement: Gas detector tubes were used to measure ammonia concentrations in the headspace of the vials.

[0163] Measurements are taken on days 2, 4, 8, 11, and 15 to track ammonia buildup overtime.

[0164] Purpose of the Experiment

[0165] Urease Activity: Urea in soil is broken down by urease into ammonia, which can be lost to the atmosphere (volatilization).

[0166] Inhibition: This test aims to see if the different NBTPT formulations inhibit urease activity, thereby reducing ammonia loss.

[0167] Time Course: Measuring ammonia over several days reveals how effective and long lasting the inhibition is.

[0168] Results: The results of the urease activity measurements of the four soil types are shown in FIG. 12A. Urease activity varied significantly between soils. Although no evaluation other than urease activity was carried out on the soils tested in this study, it was assumed that Soil 1 probably contained a higher content of urease -producing microorganisms. Soil 1, which had the highest urease activity of the four soils, was sterilized at 121°C for 20 min and urease activity was diminished significantly as shown in FIG. 12A. The soil 1 was selected for the soil test shown in FIGs. 12 B and C. FIG. 12B shows ammonia volatilities tests before encapsulation for the samples listed in Table 8, and FIG.12C shows ammonia volatilization tests after encapsulation and heat-treatment for the samples listed in Table 8.

[0169] FIG. 12 B shows the results up to day 15 of the soil test. Samples tested are listed in Table 8. Samples No. 2 and No. 3 represent the soils containing urea and heat- treated, ammonia is produced from day 2 of the test. The results indicate that heat-treating of the urea has no effect on the Urease activity. In samples No. 4 and No. 5, the NBTPT and heat-treated NBTPT were added to the soil with urea, respectively. The ammonia production were suppressed in both samples No. 4 and No. 5 up to day 4 in this condition. The behavior of heated NBTPT was similar to that of NBTPT up to day 8, but at day 15, ammonia production was predominantly reduced compared to NBTPT. The reason for the increasedstability of heated NBTPT is not clear as the activity reduction can be seen after free dying. Moreover, it was widely accepted that the NBTPT is not stable under heat-treatment. However, a mild heat-treatment at 150 °C for 5 min could be a reason for better performance by which NBTPTO may have produced and it is 100 times more effective than NBTPT.

[0170] Ammonia production at day 15 of the soil test is shown in FIG. 13. The heat- treated and mechanically mixed NBTPT / y-CD were shown to significantly reduce ammonia production. All inhibited ammonia production similarly to NBTPT. However, their behavior was the same as that of NBTPT. On the other hand, the sample treated with NBTPT and y- CD mixed in a mortar and then heat-treated at 150°C for 5 min under nitrogen inflow showed three times higher inhibition of ammonia formation compared to NBTPT.B7. Job’s method

[0171] A solution of y-CD in water (10 mM) and a solution of NBTPT in methanol (10 mM) were prepared.

[0172] Different ratio of both solutions were mixed together in test tubes, for a total volume of 1 mL, before freeze-drying.

[0173] The resulting solid was dissolved into 1 mL of D2O and analyzed by ’H NMR.

[0174] For each ratio of NBTPT (r) and for each proton (i) of NBTPT, the variation of chemical shift (Dd1) was calculated as r = [NBTPT] / ([NBTPT] + [CDa])A = - 8

[0175] For each ratio of y-CD (1 - r) and for each proton (j) of y-CD, the variation of chemical shift DdJi-rwas calculated as:A^r= ^r^0

[0176] Then r Ddi and (l-r) DdJi-r were plotted against r.

[0177] Resulting curves went through a maximum, at the r value corresponding to the ratio of the inclusion complex.

[0178] Based on Job’s method the graph shown in FIG. 14 was obtained. FIG. 14 shows that y-CD and NBTPT complex was formed as 1 : 1 complex.B8. Conclusion

[0179] Results presented in sections B1-B7 show that the complex of y-CD and NBTPT can be formed in 1 : 1 ratio and complete encapsulation can be achieved using the liquid phase preparation and freeze drying method. The capsulated NBTPT in y-CD was found to be stable at high temperature and remained stable after heating for 30 min.

[0180] NBTPT, NBTPT / y-CD capsules and NBTPT / y-CD capsules mechanical mixture all inhibited ammonia volatilization in soil in model tests. In particular, the NBTPT / y-CD capsule-machine mixture showed the highest inhibition. However, the mechanism needs to be investigated, e.g. the suppression effect is enhanced by heating.C. Procedure for Granulation

[0181] The cyclodextrin-inhibitor inclusion complex may be provided as described above. The cyclodextrin-inhibitor inclusion complex can be placed into motion and sprayed with a urea melt inside the granulator to produce the fertilizer particle. The granulated fertilizer particles generally will have a longest dimension of about 4 mm. The granulating process both fattens the cyclodextrin-inhibitor inclusion complex with urea and dries the fertilizer granules.

[0182] The spray rate of the urea melt can be controlled to control the agglomeration of fertilizer particles.

[0183] Examples of granulation process parameters that can be used are described in Table 9. Table 9D. Sample Analysis

[0184] The purity of the inhibitor, such as NBTPT and DCD, can be cross-checked by NMR, HPLC, and LCMS analysis.

[0185] Crush strength can be measured for some of the samples using a crush strength analyzer to determine the strength of the fertilizer particles.

[0186] The stability of inhibitors in the fertilizer particles can be measured using HPLC and LCMS.

[0187] The free and total moisture content of fertilizer particles can be measured using a moisture analyzer.

[0188] It is expected that the final fertilizer particles will have the following properties: crush strength (kgf): 1.68-3.60; abrasion analysis (wt. loss%): 0.11-1.50; impact resistance (shattered granules%): 0.05-1.20; moisture analysis (wt.%): 0.12-1; particle size distribution (granule): 2-4 mm (> 90%); Biuret%: 1.05-3.8; and Nitrogen%: 36.8-46.3.

[0189] The nitrogen volatilization and nitrogen transformation (nitrification) can be measured in different soils and compared to urea alone and urea with inhibitor that is not in a cyclodextrin-inhibitor inclusion complex and to products on the market such as AGROTAIN®, ESN®, and SUPERU®. A soil that is representative of a broader class of soil types can be used to measure the nitrogen volatilization and nitrification. Greenville soil and Crowley soil are two such representative soils. Other soils can also be used for the experiments described herein.

[0190] Greenville soil or Greenville clay-loam soil is typical of weathered tropical ultisols and is found in warm humid environments. The soil is classified as fine, kaolinitic, thermic Rhodic Kandiudults with a pH of 6.1-6. The soil has organic matter of 1.4%, total amount of nitrogen is about 0.06%, and the CEC is 5.2 cmol / kg. Accordingly, the soil has a low content of organic matter, and also low availability of sulfur and nitrogen. Thus, the soil is ideal for nitrogen and sulfur trials with fertilizers.

[0191] Crowley soil consists of very deep, somewhat poorly drained, very slowly permeable soils that formed in clayey fluviomarine deposits of the Pleistocene age. The soil exists in nearly level to very gently sloping soils and occurs on flat coastal plains terraces. The slope is dominantly less than 1 percent but ranges to up to 3 percent. The mean annual precipitation is about 1549 mm (61 in.), and the mean annual air temperature is about 20 degrees C (68 degrees F), where the soil is found. The soil is fine, smectitic, and thermic Typic Albaqualfs.

[0192] The nitrogen volatilization of various exemplary samples of fertilizer granules as compared to AGROTAIN®, ESN®, SUPERU®, urea, and urea with inhibitor that is not in a cyclodextrin-inhibitor inclusion complex can be determined as the percentage of nitrogen loss via ammonia volatilization as compared to the amount of nitrogen applied or as the absolute mass of nitrogen lost via ammonia volatilization. It is expected that embodiments of the fertilizer particles disclosed herein will lose less than 20 wt.% of the applied nitrogen after being exposed to soil for 20 days. It is also expected that embodiments of the fertilizerparticles disclosed herein will lose less than 20 wt.% of the applied nitrogen after being exposed to Greenville soil for 20 days and less than 20 wt.% of the applied nitrogen after being exposed to Crowley soil for 20 days. It is also expected that embodiments of the fertilizer particles disclosed herein will have lower levels of ammonia volatilization and / or nitrogen loss than AGROTAIN®, ESN®, and / or SUPERU® tested under substantially identical conditions in a given soil, which can include Greenville soil, Crowley soil, or other soils.

[0193] The stability of inhibitors, such as NBTPT, in the cyclodextrin-inhibitor inclusion complex can be monitored by measuring inhibitor concentrations in the cyclodextrin-inhibitor inclusion complex or nitrogen fertilizers containing such therein particles after different storage times. This can be performed in a controlled environment to be able to make comparisons between formulations. As demonstrated herein, it is expected that fertilizer particles with NBTPT in a cyclodextrin-inhibitor inclusion complex will have at least 90% of NBTPT remaining after storing the particles at 22° C for 30 days in a sealed container. It is expected that, within 24 hours after granulation, the fertilizer particles will have at least 95% of NBTPT remaining relative to the amount added during manufacturing. As demonstrated herein, it is expected that, on the 30th day after granulation, the fertilizer particles will have at least 90% of NBTPT remaining relative to the amount added during manufacturing. It is expected that, within 24 hours after granulation and / or on the 30th day after granulation, the weight ratio of NBTPT to all NBTPT degradation products in the fertilizer particles will be at least 10: 1. It is expected that, within 24 hours after granulation and / or on the 30th day after granulation, the weight ratio of NBTPT to n-butylamine will be at least 20: 1.

Claims

CLAIMS1. A fertilizer composition comprising: a solidified nitrogen fertilizer melt, said melt comprising one or more cyclodextrin- inhibitor inclusion complexes comprising at least one cyclodextrin and / or derivative thereof complexed with an inhibitor, wherein the inhibitor comprises at least one nitrification inhibitor and / or at least one urease inhibitor, and wherein the one or more cyclodextrin-inhibitor inclusion complexes is homogeneously dispersed throughout the solidified nitrogen fertilizer melt.

2. The fertilizer composition of claim 1, comprising: a continuous phase comprising the nitrogen fertilizer melt; and a discontinuous phase comprising the one or more cyclodextrin-inhibitor complexes dispersed throughout the continuous phase.

3. The fertilizer composition of claim 2, wherein the one or more cyclodextrin-inhibitor complexes are evenly dispersed throughout the continuous phase.

4. The fertilizer composition of any one of claims 2 to 3, wherein the discontinuous phase comprising the nitrogen fertilizer melt has a melt temperature of 140 °C or less, preferably 130 °C to 135 °C.

5. The fertilizer composition of any one of claims 1 to 3, wherein: the nitrogen fertilizer melt comprises at least 90 wt. %, preferably at least 95 wt. %, more preferably at least 98 wt. %, or even more preferably at least 99 wt. % of the fertilizer composition; and the one or more cyclodextrin-inhibitor inclusion complexes comprises up to 10 wt. %, preferably up to 5 wt. %, more preferably up to 2 wt. %, or even more preferably up to 1 wt. % of the fertilizer composition.

6. The fertilizer composition of any one of claims 1 and 3, wherein the cyclodextrin- inhibitor inclusion complexes are thermally stable at 140 °C or more and / or are stable at a pH range between 5 to 10.

7. The fertilizer composition of any one of claims 1 to 3, wherein the cyclodextrin and / or derivative thereof comprises a-cyclodextrin, P-cyclodextrin, and / or y- cyclodextrin, preferably y-cyclodextrin.

8. The fertilizer composition of any one of claims 1 to 3, wherein the nitrification inhibitor comprises 3,4-dimethylpyrazole phosphate (DMPP), thio-urea (TU), dicyandiamide (DCD), 2-chloro-6-(trichloromethyl)pyridine (nitrapyrin), 5-ethoxy-3- trichloromethyl- 1 ,2,4-thiadiazol (TERRAZOLE™), 2-amino-4-chloro-6-methyl- pyrimidine (AM), 2-mercapto-benzothiazole (MBT), ammonium thiosulfate (ATS), 2- sulfanimalamidothiazole (ST), or a combination thereof.

9. The fertilizer composition of any one of claims 1 to 3, wherein the urease inhibitor comprises N-(n-butyl)— thiophosphoric triamide, N-(n-butyl)-phosphoric triamide, N- (n-propyl) thiophosphoric triamide, benzoylthiourea (BTU), hydroquinone, acetohydroxamic acid (AHA), hydroxyurea (HU), and / or phenyl phosphorodiamidate (PPDA), preferably N-(n-butyl)-thiophosphoric triamide.

10. The fertilizer composition of any one of claims 1 to 3, wherein: the solidified nitrogen fertilizer melt is a urea fertilizer melt comprising urea; and the inhibitor is a urease inhibitor, preferably N-(n-butyl)-thiophosphoric triamide.

11. The fertilizer composition of any one of claims 1 to 3, further comprises a wetting agent, a carrier, a filler agent, and / or a binder.

12. The fertilizer composition of any one of claims 1 to 3, wherein the fertilizer composition comprises a ratio of inhibitor to degradation products of the inhibitor of 80:20 to 100:0 or wherein the fertilizer composition comprises a ratio of inhibitor comprised in the one or more cyclodextrin-inhibitor inclusion complexes to inhibitor not complexed with cyclodextrin of 80:20 to 100:0.

13. The fertilizer composition of any one of claims 1 to 3, wherein the nitrogen fertilizer melt comprises urea.

14. A method of producing the fertilizer composition of any one of claims 1 to 13, the method comprising:(a) contacting the inhibitor with a one or more cyclodextrins and / or derivatives thereof to generate a one or more cyclodextrin-inhibitor inclusion complexes; and(b) contacting the one or more cyclodextrin-inhibitor inclusion complexes with a molten nitrogen fertilizer; and(c) solidifying the molten nitrogen fertilizer to form the fertilizer composition, preferably, wherein the molten nitrogen fertilizer solubilizes the one or more cyclodextrin-inhibitor inclusion complexes such that individual molecules of the complexes are dispersed throughout the molten nitrogen fertilizer, and / or preferably, wherein steps (a) and (b) are performed as a one pot synthesis and / or multi-step synthesis and / or step (b) further comprises mixing the one or more cyclodextrin-inhibitor inclusion complexes with the molten nitrogen fertilizer.

15. A method of fertilizing, the method comprising contacting a soil, a plant, water, or a combination thereof with the fertilizer composition of any one of claims 1 to 13, preferably wherein the fertilizer composition is added directly in a solid form and / or to water before contacting the soil, plant, or soil and plant.

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