WO2019199238A1

WO2019199238A1 - Inorganic mineral entrapped in nanoparticle production method thereof

Info

Publication number: WO2019199238A1
Application number: PCT/TH2018/000015
Authority: WO
Inventors: Orawan AMNUCKSORADEJ; Pattapong KHONKHAYAN; Nattapong CHOOMKASIAN; Nitwarat RUENNARONG; Katawut NAMDEE; Teerapong YATA; Mattaka KHONGKOW; Somrudee KAEWMALUN; Warut KENGKITTIPAT
Original assignee: Vet Products Research And Innovation Center Company Limited
Priority date: 2018-04-09
Filing date: 2018-04-09
Publication date: 2019-10-17

Abstract

Inorganic mineral entrapped in nanoparticle according to this invention are comprised inorganic mineral, phospholipid, solvent, surfactant, bile salt and water. A production method of inorganic minerals entrapped in nanoparticle has the following steps. First step is preparation of oil phase solution comprising solvent homogenously mixing with phospholipid, then adding surfactant and bile salt in said solution. Second step is preparation of water phase solution by weighing inorganic minerals, water and co-surfactant. Then, said oil phase solution are homogenously mixed with said water phase solution by controlling of syringe pump. The oil phase solution is dropped into the water phase and mixed by magnetic stirrer until the final product is the solution of inorganic mineral entrapped in nanoparticle. The inorganic mineral entrapped in nanoparticle according to this invention is used the liponiosome delivery system, that having the properties to greatly store the high polarity substance or soluble substance, especially inorganic minerals. Moreover, these nanoparticles are comprised the bile salt that is an emulsifier in the animal digestive tract, to increase the stability of nanoparticles and inorganic mineral absorption ability, and to increase the production of meat, milk and egg in livestock or poultry industries.

Description

Title of invention

INORGANIC MINERAL ENTRAPPED IN NANOPARTICLE AND PRODUCTION METHOD THEREOF

Field of invention

Chemistry and nanotechnology especially in an inorganic mineral entrapped in nanoparticle and production method thereof

Background of invention

Animal feed is essential for animal livelihood and growth. Adding minerals or other essential nutrients into the animal feed that is aimed to increase the production of meat, milk or egg per the animal unit. The nutrient most filled into the animal feed as follows amino acid, vitamins or minerals such as Copper, Zinc, Iron, Manganese etc. The trace minerals are needed in small quantities, but it is necessary because they are several components in many important processes in the body such as metabolism, component of bone and teeth, blood, hormone or enzyme. Therefore, if the body is not being taken of these trace minerals, it might affect to abnormal of several body system function. In generals, minerals can be as inorganic minerals such as chloride salt, sulfate salt or oxide salt etc. Refer to US patent as publication no.US8802180 in the title of “Micronutrient supplement” disclosed that a micronutrient supplement which is made by reacting a metal oxide, or metal hydroxide, or metal carbonate of an essential mineral and an acid and/or a metal salt of an essential mineral containing a digestible binder to form an agglomerated particle of the micronutrient crystals from the slurry. The agglomerated particles provide a non-dusting, free-flowing micronutrient supplement that can be produced with desired particle sizes and densities so as to be easily mixed with a variety of feed mixtures. The digestible binder in the agglomerated particles also reduces the opportunity for interactions between the micronutrient crystals and other ingredients.

Refer to US patent with publication no. US20130035399A1 in the title of“MARKED INORGANIC ADDITIVES” disclosed inorganic mineral additive contains heavy stable isotope that can be applied in several industries including animal feed industry. However, there are some evidence that using of high quantities of inorganic minerals can be inhibited the absorption of other minerals in animal digestive system and can be reacted to Phytase enzyme, so it can reduce the capability of phosphorus releasing from the animal feed. Further, high quantities of inorganic minerals can be the pollution in environment. Using organic minerals can be solving above problems. Organic minerals is the combination of minerals and organic substance as follows amino acids, peptides, organic acid, or polysaccharides. According to The America Feed Control Officials (AAFCO), they classified organic minerals into 4 main products as follows; 1) Metal Amino Acid Complex, 2) Metal Amino Acid Chelate that the complex of salt of minerals and amino acids 1-3 molecules, preferably 2 molecules. 3) Metal Polysaccharide Complex and 4) Metal Proteinate that the product resulting from the chelation of a soluble salt with amino acids and/or partially hydrolyzed protein.

Furthermore, there are some patent that related to using organic minerals in animal feed. For example, the New Zealand patent with publication no. NZ541878A in the title of “Derivatives of seleno-amino acids with improved bioavailability and method for assuring adequate dietary requirements of selenium for livestock” disclosed organic minerals in salt form of L-seleno-alpha amino acid complex wherein the metal or mineral can be selected from Zinc, Manganese, Copper, Cobalt, or Chromium. Seleno-amino acid complex can be selected from L-seleno-methionine or Se-methyl-L-Selenocysteine. These metal L-seleno- alpha-amino acid complex salts are used as a source of selenium in animal nutrition.

Refer to Chinese patent with publication no. CN101786961B in the title of “Chromium threonine, preparation method and application thereof’ disclosed additives in the animal feed that contained chromium in organic forms with threonine amino acid resulting in chromium threonine that is organic minerals in type of metal amino acid chelate. Since, chromium combined with trivalent of threonine amino acids in the molar ratio of 0.9 - 1.1 of chromium to 3 threonine amino acids. The chromium threonine in accordance to this invention can be used as an animal feed additive and can promote synchrotron absorption of the threonine and a trace element especially chromium. It can also enhance animal growth and immunity of the animals.

Refer to Korean patent with publication no. KR1705319B1 in the title of“The feed additive composition using organic and inorganic cluster ion mineral extracting from the natural mineral and a manufacturing method thereof’ disclosed preparation of animal feed containing natural mineral extracts as the following steps (1) grinding natural minerals, mixing obtained power with inorganic acid and water, stirring the mixture, and filtering (2) adding amino acid solution to obtained water-soluble complex ionic mineral and performing metal chelating reaction (3) adding organic acid to obtained solution, adding water dispersed nano-sulfur power into the obtained mixture, and (4) mixing obtained stabilized water- dispersed nano-sulfur sol in feed materials, drying in a dryer, and grinding and processing into a powder. According to above step, all supplement components of invention contained both inorganic acid and organic acid in the animal feed.

However, organic minerals are high cost, so it will be added especially in high-graded animal feed or only special request from customer. Therefore, using of organic minerals are limitation in some special consumers. Furthermore, most of animal feed producers still using inorganic minerals.

As mentioned above, gastrointestinal epithelium is physical barrier of inorganic mineral absorption into blood circulatory system. In the present, there are some researches in related to development of drug delivery system via several carriers to increase the capability of nutrient absorption into the blood stream, to reduce the problem of using drug in overdose, and contamination of chemical in the environment.

The carriers in drug delivery can be prepared from several biodegradable materials such as lipids-based materials. The carriers need to the basic properties as stable, non-toxicity to increase effective and safety of contained drug. In the present, the lipid-based nanoparticles are greatly well-known such as synthetic lipids or natural lipids to be used as the drug delivery carriers to increase bioavailability of the drugs, controlled released and nutrient absorption efficacy of gastrointestinal epithelium by increasing drug dissolution, and promote gastric emptying rate and longer drug releasing. Moreover, lipid can be enhanced lymph production and circulation wherein the contained drug within lipid-based nanoparticles can be simultaneous absorbed with lipid absorption that it is called“Trojan horse strategy”.

Considering to patent related to drug delivery system in nanoparticles form using in animal feed, there is Korean patent with publication no. KR10163298B1 in the title of “A manufacturing method for nanoliposome liquid feedstuff and liquid feed stuff manufactured by the same” disclosed nanoliposome liquid feed stuff for animal preparation especially in livestock, poultry that contained the main component as follows; hypochlorus acid, ion silicate, beta-glucan, bentonite, zeolite, illite, neem oil, distilled water and liquid sulfur. First, the above components are grinded into 30 - 100 meshes to perform a mucus product. Then, the mixture is grinded into 600 - 800 meshes. Next step is a pretreatment step of forming a purified product by firstly removing impurities by agitating the mucus product and secondly removing impurities after setting the impurities; a liposome step of forming a vesicle by mixing the purified product with a phospholipid mixture and agitating and drying the purified product to saturate particles of the purified product in the phospholipid and to capsulate the phospholipid. Next step is a post-treatment step of completing a nanoliposome composition by agitating a mixture obtained by mixing 5 - 10 wt% of the purified product with 90 - 95 wt% of distilled water to disperse the purified product and agitating the mixture at 100-200 rpm for 30 - 60 minutes.

Nanoparticles or liponiosomes are mostly used as the drug delivery carriers including animal feed industries. Because, the liponiosomes contains soluble active ingredients that structural based of liposome are phospholipid bilayer and non-ionic surfactant as the particle wall, so these particles can greatly store only high polarity or soluble ingredients. However, the limitation of these particles is instability especially in acid environment in digestive tract.

Therefore, inorganic minerals entrapped in nanoparticles in accordance with this invention has been developed using bile salt, wherein has an emulsifier in the digestive system, becomes part of ingredients of the liponiosome wall to entrap the inorganic mineral in the animal feed. Bile salt contains hydrophilic side and hydrophobic side, A typical micelle forms an aggregrate with the hydrophobic side in the micelle center, while the hydrophilic side arranged into the outside that contact with surrounding solvent. Arrangement of micelle structure is the benefit to use as the main component of the lipid-based nanoparticle. Moreover, the bile salt should be Sodium glycolate and sodium cholate that are the main functions in fat or oil digestion in small intestine by reducing the size of lipid droplet to increase digestive surface area, so it affects more efficiency of lipid digestion and absorption.

Summary of invention Inorganic mineral entrapped in nanoparticle according to this invention are comprised inorganic mineral, phospholipid, solvent, surfactant, bile salt and water. A production method of inorganic minerals entrapped in nanoparticle has the following steps. First step is preparation of oil phase solution comprising solvent homogenously mixing with phospholipid, then adding surfactant and bile salt in said solution. Second step is preparation of water phase solution by weighing inorganic minerals, water and co-surfactant. Then, said oil phase solution are homogenously mixed with said water phase solution by controlling of syringe pump with flow rate of 0.1 - 1.0 ml per minute. The oil phase solution is dropped into the water phase and mixed by magnetic stirrer with speed of 300 - 500 rounds per minute (RPM) until the final product is the solution of inorganic mineral entrapped in nanoparticle.

Inorganic mineral entrapped in nanoparticle according to this invention is used the liponiosome delivery system, that having the properties to greatly store the high polarity substance or soluble substance, especially inorganic minerals. Moreover, these nanoparticles are comprised the bile salt that is an emulsifier in the animal digestive tract, to increase the stability of nanoparticles and inorganic mineral absorption ability, and to increase the production of meat, milk and egg in livestock or poultry industries.

Brief description of the drawings

FIG.l illustrates stability comparison results of inorganic mineral entrapped in nanoparticle solution in different temperature FIG.2 illustrates stability comparison results from adding bile salt that is the main ingredient of inorganic mineral entrapped in nanoparticle (FIG.2a) illustrates appearance of nanoparticle solution without bile salt and (FIG.2b) illustrates appearance of nanoparticle solution with bile salt. Detailed description of the invention

Inorganic mineral entrapped in nanoparticle in accordance with this invention are comprised

Inorganic mineral 0.5 - 5 % by weight

Phospholipid 0.5 - 2 % by weight

Solvent 25 - 35 % by weight

Surfactant 0.1 - 0.6 % by weight

Co-surfactant 0.1 - 0.6 % by weight

Bile salt 0.01 - 0.08 % by weight

Water 55 - 70 % by weight

The inorganic mineral can be selected from metal or non-metal or combination thereof, preferably metal. The metal can be selected from copper, manganese, zinc, iron, cobalt, chromium and combination thereof.

The solvent can be selected from alcoholic compound or glycol or combination thereof, such as ethyl alcohol or ethoxydiglycol or propylene glycol or butylene glycol or dipropylene glycol or combination thereof, preferably ethyl alcohol.

The surfactant can be selected from ionic surfactant or non-ionic surfactant or combination thereof, preferably the non-ionic surfactant. The non-ionic surfactant can be selected from Sorbitan monopalmitate or Sorbitan monooleate or Sorbitan monolaurate or combination thereof, preferably Sorbitan monooleate.

The co-surfactant can be selected from Polyoxyethylene (20) sorbitan monooleate or Polyoxyethylene (20) sorbitan monostearate or Polyoxyethylene (20) sorbitan monopalmitate or Polyoxyethylene (20) sorbitan monolaurate or combination thereof, preferably Polyoxyethylene (20) sorbitan monolaurate. The bile salt can be selected from at least one of the following: bile acid compounds from cholesterol or combination thereof such as cholic acid or chenodeoxycholic acid or lithocholic acid or deoxycholic acid or taurocholic acid or glycocholic acid or taurochenodeoxycholic acid or glycochenodeoxycholic acid or combination thereof. The most preferably bile salt is deoxycholic acid.

The inorganic mineral entrapped in nanoparticle is provided different of the optimum ratio in one embodiment of composition of inorganic mineral entrapped in nanoparticle are shown subsequently wherein the embodiments do not limit the chemicals and other ratios which are known to the skilled person in the related arts that these chemicals have similar result, and by which any improvements or modifications may be possible within the claims according to this invention.

Example 1 Composition for synthesizing inorganic mineral entrapped in nanoparticle comprised the following:

Inorganic mineral 3 % by weight

phospholipid 1 % by weight

solvent 28.5 % by weight

surfactant 0.5 % by weight

co-surfactant 0.5 % by weight

bile salt 0.054 % by weight

water 66.5 % by weight

Example 2 Composition for synthesizing inorganic mineral entrapped in nanoparticle comprised the following:

Inorganic mineral 0.5 % by weight

phospholipid 1 % by weight

solvent 28.5 % by weight

surfactant 0.5 % by weight

co-surfactant 0.5 % by weight

bile salt 0.054 % by weight

water 69.0 % by weight

Example 3 Composition for synthesizing inorganic mineral entrapped in nanoparticle comprised the following: Inorganic mineral 1.5 % by weight

phospholipid 1 % by weight

solvent 28.5 % by weight

surfactant 0.5 % by weight

co-surfactant 0.5 % by weight

bile salt 0.054 % by weight

water 59.0 % by weight

A production method of inorganic mineral entrapped in nanoparticle according to this invention are comprised the following steps; a. Preparation of oil phase solution: weighing the phospholipid in the range of 0.5 - 2 % by weight, preferably 1 % by weight, and weighing the solvent in the range of 25 - 35 % by weight, preferably 28.5 % by weight, then adding the phospholipid into the solvent and mixing homogenously, then adding the surfactant in the range of 0.1 - 0.6 % by weight, preferably 0.5 % by weight and the bile salt in the range of 0.01 - 0.08 % by weight, preferably 0.054 % by weight, and mixing whole solution homogenously. The product of this step called oil phase solution. b. Preparation of water phase solution: weighing the inorganic mineral in the range of 0.5 - 5 % by weight, preferably 3 % by weight, then dissolving into the water, then adding the co-surfactant in the range of 0.1 - 0.6 % by weight, preferably 0.5 % by weight, and mixing whole solution homogenously.

c. Mixing both phase solutions: the oil phase solution is homogenously mixed with the water phase solution by controlling of syringe pump with flow rate of 0.1 - 1.0 ml per minute, preferably 0.8 ml per minute. The oil phase solution is dropped into the water phase and mixed by magnetic stirrer with speed of 300 - 500 rounds per minute (RPM) until the final product is the solution of inorganic mineral entrapped in nanoparticle.

The inorganic mineral according to above production method can be selected from one component metal or non-metal or combination thereof, preferably metal. The metal can be selected from at least one of copper, manganese, zinc, iron, cobalt, chromium or combination thereof. The solvent according to above production method can be selected from alcoholic compound or glycol or combination thereof, such as ethyl alcohol or ethoxydiglycol or propylene glycol or butylene glycol or dipropylene glycol or combination thereof, preferably ethyl alcohol.

The surfactant according to above production method can be selected from ionic surfactant or non-ionic surfactant or combination thereof, preferably the non-ionic surfactant. The non-ionic surfactant can be selected from Sorbitan monopalmitate or Sorbitan monooleate or Sorbitan monolaurate or combination thereof, preferably Sorbitan monooleate.

The co-surfactant according to above production method can be selected from Polyoxyethylene (20) sorbitan monooleate or Polyoxyethylene (20) sorbitan monostearate or Polyoxyethylene (20) sorbitan monopalmitate or Polyoxyethylene (20) sorbitan monolaurate or combination thereof, preferably Polyoxyethylene (20) sorbitan monolaurate.

The bile salt according to above production method can be selected from at least one of the following: bile acid compounds from cholesterol or combination thereof such as cholic acid or chenodeoxycholic acid or lithocholic acid or deoxycholic acid or taurocholic acid or glycocholic acid or taurochenodeoxycholic acid or glycochenodeoxycholic acid or combination thereof. The most preferably bile salt is deoxycholic acid.

Then, the inorganic mineral entrapped in nanoparticle is obtained to test the stability as shown the following results;

Example 4 Considerations of particle stability based on hydrodynamic diameter and zeta potential

Through the analysis of the hydrodynamic diameter and zeta potential, together with the physical characteristics of the alternative accelerated conditions and accelerated conditions for 1 month at 25 and 45 degrees Celsius, the test results showed that the storage temperature of inorganic mineral entrapped in nanoparticle at 45 degrees Celsius will change the hydrodynamic diameter significantly and color appearance as shown in FIG.l. However, it is not occurred precipitate or layer separation While, the storage temperature at 25 degrees Celsius that shown the great stability, acceptable of zeta potential and applicable to use for further researches. Table 1 The hydrodynamic diameter and the zeta potential of the sample of the inorganic mineral entrapped in nanoparticle for 1 -month period.

Inorganic mineral entrapped in nanoparticle using liponiosome according to this invention can be further developed or incorporated with other methods to derive any appropriate forms - such as in liquid, solid or powdered forms or any other forms of choices with no limitations.

Example 5 stability comparison results from adding bile salt that is the main ingredient of inorganic mineral entrapped in nanoparticle

Bile salt have the properties to increase the nanoparticle stability according to this invention, wherein there is not any precipitate in the solution with adding the bile salt into the system under the optimal condition as shown in FIG.2a. The solution without the bile salt can not be formed the particle in nanometer size, but larger that cannot be measured by nanosizer, while there is the precipitate and layer separation of the solution after leaving for a period to study the stability as shown in FIG.2b. Although this invention has been disclosed in the context of certain embodiment and examples, it will be understood by those skilled in the art that the present invention extends beyond the specifically disclosed embodiment to other alternative embodiments which any improvements or modifications may be possible within the claims according to this invention.

Claims

1. An inorganic mineral entrapped in nanoparticle are comprising component as follows;

Inorganic mineral 0.5 - 5 % by weight

Phospholipid 0.5 -2 % by weight

Solvent 25 - 35 % by weight

Surfactant 0.1 - 0.6 % by weight

Co-surfactant 0.1 - 0.6 % by weight

Bile salt 0.01 - 0.08 % by weight

Water 55 - 70 % by weight

2. The inorganic mineral entrapped in nanoparticle according to claim 1 wherein the inorganic mineral can be selected from at least one of metal or non-metal or combination thereof.

3. The inorganic mineral entrapped in nanoparticle according to claim 1 or 2 wherein the inorganic mineral is preferably metal.

4. The inorganic mineral entrapped in nanoparticle according to eithers of claim 1 to 3 wherein the metal can be selected from at least one of copper, manganese, zinc, cobalt or chromium or combination thereof.

5. The inorganic mineral entrapped in nanoparticle according to claim 1 wherein the solvent can be selected from at least one of alcoholic compound or glycol or combination thereof.

6. The inorganic mineral entrapped in nanoparticle according to claim 1 or 5 wherein the solvent can be selected from ethyl alcohol or ethoxy glycol or propylene glycol or butylene glycol or dipropylene glycol or combination thereof.

7. The inorganic mineral entrapped in nanoparticle according to claim 1 wherein the surfactant can be selected from at least one of ionic surfactant or non-ionic surfactant or combination thereof.

8. The inorganic mineral entrapped in nanoparticle according to claim 1 or 7 wherein the surfactant is preferably non-ionic surfactant that can be selected from at least one of sorbitan monopalmitate or sorbitan onooleate or sorbitan onolaurate or combination thereof.

9. The inorganic mineral entrapped in nanoparticle according to claim 1 wherein the cosurfactant can be selected from at least one of Polyoxyethylene (20) sorbitan monooleate or Polyoxyethylene (20) sorbitan monostearate or Polyoxyethylene (20) sorbitan monopalmitate or Polyoxyethylene (20) sorbitan monolaurate or combination thereof,

10. The inorganic mineral entrapped in nanoparticle according to claim 1 wherein bile salt can be selected from at least one of bile acid from cholesterol or combination of said bile acid thereof.

11. The inorganic mineral entrapped in nanoparticle according to claim 10 wherein the bile acid can be selected from cholic acid or chenodeoxycholic acid or lithocholic acid or deoxycholic acid or taurocholic acid or glycocholic acid or taurochenodeoxycholic acid or glycochenodeoxycholic acid or combination thereof.

12. The inorganic mineral entrapped in nanoparticle according to claim 10 or 11 wherein the bile acid is most preferably deoxycholic acid.

13. A production method of inorganic mineral entrapped in nanoparticle are comprised the following steps;

a. Preparation of oil phase solution: weighing the phospholipid in the range of 0.5 - 2 % by weight and weighing the solvent in the range of 25 - 35 % by weight, then adding the phospholipid into the solvent and mixing homogenously, then adding the surfactant in the range of 0.1 - 0.6 % by weight and the bile salt in the range of 0.01 - 0.08 % by weight, and mixing whole solution homogenously;

b. Preparation of water phase solution: weighing the inorganic mineral in the range of 0.5 - 5 % by weight, then dissolving into the water, then adding the co-surfactant in the range of 0.1 - 0.6 % by weight and mixing whole solution homogenously;

c. Mixing both phase solutions: the oil phase solution is homogenously mixed with the water phase solution by controlling of syringe pump with flow rate of 0.1 - 1.0 ml per minute, the oil phase solution is dropped into the water phase and mixed by magnetic stirrer with speed of 300 - 500 rounds per minute (RPM) until the final product is the solution of inorganic mineral entrapped in nanoparticle.

14. The production method of inorganic mineral entrapped in nanoparticle according to claim 13 wherein the inorganic mineral can be selected from at least one of metal or non-metal or combination thereof.

15. The production method of inorganic mineral entrapped in nanoparticle according to claim 13 or 14 wherein the inorganic mineral is preferably metal.

16. The production method of inorganic mineral entrapped in nanoparticle according to either of claims 13 to 15 wherein the metal can be selected from at least one of copper, manganese, zinc, cobalt or chromium or combination thereof.

17. The production method of inorganic mineral entrapped in nanoparticle according to either of claims 13 to 16 wherein the inorganic mineral is preferably concentration of 3% by weight.

18. The production method of inorganic mineral entrapped in nanoparticle according to claim 13 wherein the solvent can be selected from at least one of alcoholic compound or glycol or combination thereof.

19. The production method of inorganic mineral entrapped in nanoparticle according to claim 13 or 18 wherein the solvent can be selected from ethyl alcohol or ethoxy glycol or propylene glycol or butylene glycol or dipropylene glycol or combination thereof.

20. The production method of inorganic mineral entrapped in nanoparticle according to either of claim 13 or 18 or 19 wherein the solvent is preferably in concentration of 28.5 % by weight.

21. The production method of inorganic mineral entrapped in nanoparticle according to either of claims 13 wherein the surfactant can be selected from at least one of ionic surfactant or non-ionic surfactant or combination thereof.

22. The production method of inorganic mineral entrapped in nanoparticle according to claim 13 or 21 wherein the surfactant is preferably non-ionic surfactant that can be selected from at least one of sorbitan monopalmitate or sorbitan monooleate or sorbitan monolaurate or combination thereof.

23. The production method of inorganic mineral entrapped in nanoparticle according to either of claims 13 or 21 or 22 wherein the surfactant is preferably in concentration of 0.5 % by weight.

24. The production method of inorganic mineral entrapped in nanoparticle according to claim 13 wherein the co-surfactant can be selected from at least one of Polyoxyethylene (20) sorbitan monooleate or Polyoxyethylene (20) sorbitan monostearate or Polyoxyethylene (20) sorbitan monopalmitate or Polyoxyethylene (20) sorbitan monolaurate or combination thereof,

25. The production method of inorganic mineral entrapped in nanoparticle according to claim 13 or 24 wherein the co-surfactant is preferably in concentration of 0.5 % by weight.

26. The production method of inorganic mineral entrapped in nanoparticle according to claim 13 wherein the phospholipid is preferably in concentration of 1 % by weight.

27. The production method of inorganic mineral entrapped in nanoparticle according to claim 13 wherein the bile salt is preferably in concentration of 0.054 % by weight.

28. The production method of inorganic mineral entrapped in nanoparticle according to claim

3 wherein the flow rate of syringe pump is preferably at 0.8 ml per minute.