Claims:1. A method of synthesizing biotin from biofortified raw material, comprises:
taking a raw material (101), wherein the raw material is biofortified dried sesbania grandiflora leaves;
crushing the raw material to obtain a powder (102);
carrying out an extraction of the raw material (103), wherein the extraction is carried out by circulating the raw material and an aqueous solution of ethanol in a ratio to an extractor at a temperature for atleast three hours, wherein the extraction is carried out for atleast two times and filtrates are combined;
undergoing a process (1); and
undergoing a process (2), wherein the process (1) and process (2) are carried out separately and simultaneously (104); and
combining dried powders obtained from process (1) and process (2) (105);
milling the powder (106);
blending the powder for an hour (107);
sifting the powder (108); and
packing the powder (109).
2. The method as claimed in claim 1, wherein the process (1) comprises the steps of:
passing the filtrates through a membrane (201);
collecting a filtrate and a supernatant separately (202);
concentrating the supernatant to obtain a concentrated supernatant (203), wherein the concentrated supernatant has a concentration of 75 to 80%;
passing the filtrate through a column packed with agarose polyaldehyde beads (204);
eluting the column with 9% lemon juice (205);
collecting the eluate (206);
concentrating the eluate at 70°C to 80°C to obtain a concentrated solution having a TDS of 30 to 40% (207);
spray drying the concentrated solution from column elution and concentrated supernatant from membrane filtration to obtain a powder (208).
3. The method as claimed in claim 1, wherein the process (2) comprises the steps of:
concentrating the filtrate solution (301);
precipitating the filtrate solution by acidification (302), wherein the acidification is done using a lemon juice;
allowing the precipitates to settle down in the solution overnight (303);
filtering the solution and collecting a supernatant (304);
spray drying the solution to obtain a powder (305).
4. The method as claimed in claim 1, wherein the ratio of the raw material and the aqueous solution of ethanol is 1:3.
5. The method as claimed in claim 2 and 3, wherein the spray drying is done at an inlet temperature 180° to 190°C and outlet temperature of 80°C to 100°C.
6. The method as claimed in claim 1, wherein the sifting is done through a 40 size mesh or sieve.
7. The method as claimed in claim 1, wherein the extraction is carried out at a temperature in the range of 70°C to 80°C.
8. The method as claimed in claim 2, wherein the filtrate is passed through a membrane having a pore size of 0.03 to 10 microns with a feed operating pressure of approximately 100 to 400kPa.
, Description:A) TECHNICAL FIELD OF INVENTION
[001] The present invention generally relates to multivitamin and mineral supplement composition and particularly to a method of synthesizing enriched vitamins and minerals from biofortified raw materials. The present invention more particularly relates to a composition and a method of synthesizing natural vitamins Band enriched using agronomic biofortified crops for use in intestinal inflammation associated with dysbiosis of the gut microbiota.

B) BACKGROUND OF INVENTION
[002] The gut microbiota maintains a symbiotic relationship with the host and regulates several important functions including host metabolism, immunity, and intestinal barrier function. Intestinal inflammation is commonly associated with dysbiosis of the gut microbiota. Alterations in the gut microbiota and associated changes in metabolites as well as disruptions in the intestinal barrier are evidence of the relationship between the gut microbiota and intestinal inflammation. Recent studies have found that many factors may alter the gut microbiota, with the effects of diet being commonly studied.
[003] There are number of ways wherein an unhealthy gut might manifest itself. Some of the most common signs are upset stomach, high sugar diet, unintentional weight changes, sleep disturbances and constant fatigue, skin irritation, autoimmune conditions, food intolerances, etc. Diet is one of many important factors which can affect the gut microbiota and many other aspects of human health and disease. The gut microbiota uses ingested nutrients as an energy source in fundamental biological processes, and changes in diet type or pattern may change the composition of the gut microbiota, as species more suited to metabolizing novel diet types will increase in population, while other species may become less abundance. These changes in the composition of the gut microbiota may affect host physiology and disease resistance.
[004] Fortification is the practice of deliberately increasing the content of an essential micronutrient, i.e. vitamins and minerals (including trace elements) in a food, so as to improve the nutritional quality of the food supply and provide a public health benefit with minimal risk to health. Biofortification is the process by which the nutritional quality of food crops is improved through agronomic practices, conventional plant breeding, or modern biotechnology. Biofortification differs from conventional fortification in that biofortification aims to increase nutrient levels in crops during plant growth rather than through manual means during processing of the crops. Biofortification may therefore present a way to reach populations where supplementation and conventional fortification activities may be difficult to implement and/or limited.
[005] The agronomical techniques include the application of various abiotic elicitors, precursor molecules, pathway intermediates, zinc containing fertilizers, use of soil amendments like lime, gypsum, organic amendments like fym, application of bio-fertilizers like rhizobium (for legumes), azotobacter and azospirillum (non-leguminous crops) and azolla (water fern). Biofortification of curry leaves, holy basil leaves, guava fruits, guava leaves, annato seeds, lemon fruits, amla fruits, Sesbania grandiflora, and bamboo shoots are being employed commercially.
[006] However, there is still a need for a nutritional composition that is capable of promoting a well-balanced intestinal microbiota. There is a need to develop a composition and a method of synthesizing a composition or products that are enriched with essential vitamins and minerals which promote the growth of the gut microbiota and promote or restore an optimal intestinal microbiota ecosystem in an individual beneficial for it. There is a need to come up with a method of synthesizing enriched vitamin B wherein the vitamin is biotin from biofortified raw materials.
[007] The value additions and above mentioned shortcomings, disadvantages and problems are addressed herein, as detailed below.

C) OBJECT OF INVENTION
[008] The primary object of the present invention is to provide a method of synthesizing enriched biotin from biofortified raw materials.
[009] Another object ofthe present invention is to provide a multivitamin and mineral composition used to promote the growth of the gut microbiota and to promote or restore an optimal intestinal microbiota ecosystem in an individual beneficial for it.
[0010] Yet another object of the present invention is to provide a multivitamin and mineral composition which is in the form of a powder.
[0011] Yet another object of the present invention is to provide a novel method of synthesizing the multivitamin and mineral composition, wherein the vitamin is biotin.
[0012] These and other objects and advantages of the embodiments herein will become readily apparent from the following detailed description taken in conjunction with the accompanying drawings.

D) SUMMARY OF INVENTION
[0013] The embodiments of the present invention provide amethod of synthesizing biotin from enriched raw materials. The raw material is Biofortified dried sesbania grandiflora leaves.
[0014] According to an embodiment of the present invention, amethod of synthesizing biotin from biofortified raw material comprisestaking a raw material (101). The raw material is crushed to obtain a powder (102). An extraction of the raw material is carried out (103). Then, a process (1) is undergone and a process (2) is undergone, separately and simultaneously (104). The dried powders obtained from process (1) and process (2) are combined to obtain a final powder (105). The powder is milled (106). The powder is blended for an hour (107). The powder is sifted (108). The powder is packed (109).
[0015] According to an embodiment of the present invention, the raw material is biofortified dried sesbania grandiflora leaves.
[0016] According to an embodiment of the present invention, the extraction is carried out by circulating the raw material and an aqueous solution of ethanol in a ratio to an extractor at a temperature for atleast three hours, wherein the extraction is carried out for atleast two times and filtrates are combined. The ratio of the raw material and the aqueous solution of ethanol is 1:3.
[0017] According to an embodiment of the present invention, the process (1) comprises the steps ofpassing the filtrates through a membrane (201). The filtrate is passed through a membrane having a pore size of 0.03 to 10 microns with a feed operating pressure of approximately 100 to 400kPa.The filtrate and the supernatant is collected separately (202). The supernatant is concentrated to obtain a concentrated supernatant having a percentage of 75 to 80% (203). The filtrate is passed through a column packed with agarose polyaldehyde beads (204). The column is eluted with a 9% lemon juice (205). The eluate is collected (206). The eluate is concentrated at 70°C to 80°C to obtain a concentrated solution having a TDS of 30 to 40% (207). The concentrated solution obtained from the column elution and the concentrated supernatant from membrane filtration is spray dried to obtain a powder (208).
[0018] According to an embodiment of the present invention, the process (2) comprises the steps ofconcentrating the filtrate solution (301). The filtrate solution is precipitated by acidification (302). The acidification is done using a lemon juice. The precipitates are allowed tosettle down in the solution overnight (303). The solution is filtered and a supernatant collected (304). The solution is spray dried to obtain a powder (305).
[0019] According to an embodiment of the present invention, the spray drying is done at an inlet temperature 180° to 190°C and outlet temperature of 80°C to 100°C.
[0020] According to an embodiment of the present invention, the sifting is done through a 40 size mesh or sieve.
[0021] According to an embodiment of the present invention, the extraction is carried out at a temperature in the range of 70°C to 80°C.
[0022] These and other aspects of the embodiments herein will be better appreciated and understood when considered in conjunction with the following description and the accompanying drawings. It should be understood, however, that the following descriptions, while indicating preferred embodiments and numerous specific details thereof, are given by way of illustration and not of limitation. Many changes and modifications may be made within the scope of the embodiments herein without departing from the spirit thereof, and the embodiments herein include all such modifications.

E) BRIEF DESCRIPTION OF DRAWINGS
[0023] The other objects, features and advantages will occur to those skilled in the art from the following description of the preferred embodiment and the accompanying drawings in which:
[0024] FIG. 1 is a flowchart showing the steps involved in the method of synthesizing biotin from biofortified raw material, according to an embodiment of the present invention.
[0025] FIG. 2 is a flowchart showing the steps involved in the method of membrane filtration [process (1)] of the filtrate obtained after the extraction process of the raw material, according to an embodiment of the present invention.
[0026] FIG. 3 is a flowchart showing the steps involved in the process (2) undergone by the filtrate obtained after the extraction process of the raw material, according to an embodiment of the present invention.
[0027] FIG. 4 is a schematic showing the synthesis of biotin, according to an embodiment of the present invention.

F) DETAILED DESCRIPTION OF DRAWINGS
[0028] In the following detailed description, a reference is made to the accompanying drawings that form a part hereof, and in which the specific embodiments that may be practiced is shown by way of illustration. The embodiments are described in sufficient detail to enable those skilled in the art to practice the embodiments and it is to be understood that the logical, mechanical, electronic and other changes may be made without departing from the scope of the embodiments. The following detailed description is therefore not to be taken in a limiting sense.
[0029] The various embodiments of the present invention provide a multivitamin and mineral compositioncomprising natural vitamins and minerals extracted and enriched using agronomic biofortified crops. The composition is used is situation of intestinal inflammation associated with dysbiosis of the gut microbiota. According to the embodiments of the present invention, the vitamin is biotin vitamin.
[0030] FIG. 1 is a flowchart showing the steps involved in the method of synthesizing biotin from biofortified raw material, according to an embodiment of the present invention. With respect to FIG. 1, the method comprisestaking a raw material (101). The raw material is crushed to obtain a powder (102). An extraction of the raw material is carried out (103). Then, a process (1) is undergone and a process (2) is undergone, separately and simultaneously (104). The dried powders obtained from process (1) and process (2) are combined to obtain a final powder (105). The powder is milled (106). The powder is blended for an hour (107). The powder is sifted (108). The powder is packed (109).
[0031] According to an embodiment of the present invention, the raw material is biofortified dried sesbania grandiflora leaves.
[0032] According to an embodiment of the present invention, the extraction is carried out by circulating the raw material and an aqueous solution of ethanol in a ratio to an extractor at a temperature for atleast three hours, wherein the extraction is carried out for atleast two times and filtrates are combined. The ratio of the raw material and the aqueous solution of ethanol is 1:3.
[0033] FIG. 2 is a flowchart showing the steps involved in the method of membrane filtration [process (1)] of the filtrate obtained after the extraction process of the raw material, according to an embodiment of the present invention. With respect to FIG. 2, the process (1) comprises the steps of passing the filtrates through a membrane (201). The filtrate is passed through a membrane having a pore size of 0.03 to 10 microns with a feed operating pressure of approximately 100 to 400kPa.The filtrate and the supernatant is collected separately (202). The supernatant is concentrated to obtain a concentrated supernatant having a concentration of 75 to 80% (203); The filtrate is passed through a column packed with agarose polyaldehyde beads (204). The column is eluted with a 9% lemon juice (205). The eluate is collected (206). The eluate is concentrated at 70°C to 80°C to obtain a concentrated solution having a TDS of 30 to 40% (207). The concentrated solution obtained from the column elution and the concentrated supernatant from membrane filtration is spray dried to obtain a powder (208).
[0034] FIG. 3 is a flowchart showing the steps involved in the process (2) undergone by the filtrate obtained after the extraction process of the raw material, according to an embodiment of the present invention. With respect to FIG. 3, the process (2) comprises the steps ofconcentrating the filtrate solution (301). The filtrate solution is precipitated by acidification (302). The acidification is done using a lemon juice. The precipitates are allowed tosettle down in the solution overnight (303). The solution is filtered and a supernatantis collected (304). The solution is spray dried to obtain a powder (305).
[0035] According to an embodiment of the present invention, the spray drying is done at an inlet temperature 180° to 190°C and outlet temperature of 80°C to 100°C.
[0036] According to an embodiment of the present invention, the sifting is done through a 40 size mesh or sieve.
[0037] According to an embodiment of the present invention, the extraction is carried out at a temperature in the range of 70°C to 80°C.

PROCESS AND PROCEDURE IN DETAIL
[0038] FIG. 4 is a schematic showing the synthesis of biotin, according to an embodiment of the present invention. With respect to FIG. 4:
[0039] Raw material: Biofortified dried sesbania grandiflora leaves free from foreign material are selected.
[0040] Crushing: Crush the dried leaves to reduce the size to 8mm - 10mm using hammer mill.
[0041] Extraction: Charge the raw material and aqueous ethanol in the ratio 1:3, intothe extractor. Raise the temperature to 70°C to 80°C and circulate the water for 3 hours. After 3 hour filter the extract and collect the filtrate.
[0042] Repeat the above extraction step for one more time. Combine both the filtrates.
[0043] The filtrates obtained from extraction will undergo two types of process:
[0044] Process 1:
[0045] 1) Membrane filtration: Pass the filtrates through membrane with pore size 0.03 to 10 microns, with feed operating pressure approximately 100 to 400kPa (15 to 60psi). Collect both Filtrate and supernatant separately. Concentrate the supernatant.
[0046] 2) Column elution/separation: Pass the filtrate through the column packed with agarose polyaldehyde beads (resin). Elute the column with 9% lemon juice. Collect the eluate and concentrate at 70°C to 80°C to obtain TDS of 30 to 40%.
[0047] 3) Spray drying: Spray dry the concentrated solution from column elution and concentrated supernatant from membrane filtration at inlet temperature 180° to 190°C and outlet temperature 80°C to 100°C. Collect the dried powder.
[0048] Process 2:
[0049] 1) Concentration: Concentrate the filtrates obtained from extraction.
[0050] 2) Acidification: precipitate the filtrate by acidification using lemon juice.
[0051] 3) Settling: After acidification, allow the filtrate for settling overnight.
[0052] 4) Filtration: Filter the above solution and collect the supernatant.
[0053] 5) Spray dry : Spray dry the solution at inlet temperature 180° to 190°C and outlet temperature 80°C to 100°C. Collect the dried powder.
[0054] Packing : Combine dried powder from the process 1 and 2 are milled, blended an hour, sifted through 40mesh and packed in a air tight container.
[0055] According to an embodiment of the present invention, the biotin synthesized is effective in the treatment of diarrhoea, irritable Bowel Syndrome (IBS), antibiotic associated diarrhoea (AAD) related to Clostridium difficile, Clostridium difficile associated diarrhoea (CDAD) and recurrent Clostridium difficile infections (RCDI).
[0056] According to an embodiment of the present invention, the dietary supplement further comprises dietary supplemental amounts of one or more compounds selected from vitamins and minerals other than calcium or phosphorus. The supplement is biotin.

G) ADVANTAGES OF INVENTION
[0057] The present invention addresses the problem of gut dysbiosis wherein the composition of the present invention assists in promoting a gut lining and tight junction functioning, ultimately leading to associated effects like reduced inflammation in the intestinal lining.
[0058] The biotin synthesized is used for improving the condition of the digestive system by maintaining a healthy microbiota population by mending ulcerated and damaged intestinal epithelial cell surfaces for the microbiota to populate.
[0059] The biotin in combination with vitamins and minerals supplies the right amount of the right micronutrients at the right time to assure adequate intake of micronutrients needed for disease prevention and protection against nutritional losses and deficiencies due to lifestyle factors and common inadequate dietary patterns.
[0060] It is to be understood that the phraseology or terminology employed herein is for the purpose of description and not of limitation. Therefore, while the embodiments herein have been described in terms of preferred embodiments, those skilled in the art will recognize that the embodiments herein can be practiced with modification within the spirit and scope of the claims presented in the complete specification or non-provisional application.