DESC:Stabilized Vitamin K Compositions
Related application:
This application claims the priority of the provisional application number 201621005176 dated 15thFebruary 2016.

Field of the invention:
The present invention relates to compositions comprising Vitamin K and pharmaceutically acceptable excipients and / or a mixture of such pharmaceutically acceptable excipients which stabilize Vitamin K against degradation in the presence of moisture, in the presence of alkaline media, and in the presence of light. The compositions so prepared are useful in the preparation of a wide variety of products/dosage forms, more specifically tablets, capsules, sachets and emulsions in which Vitamin K is stabilized against the action of light, moisture and alkaline pH for 24 months minimum.

Background of the Invention
Vitamin K was discovered in the early twentieth century wherein it was shown to play a critical role in blood coagulation. Since then Vitamin K has been found to play a crucial role in many aspects of human health.
Vitamin K refers to a group of compounds, which share a methylated naphthoquinone ring structure in common. The compounds bear a lipophilic side chain at the 3-position of the 2-methyl-I, 4-naphthoquinone ring structure. The side chain in phylloquinone (Vitamin K1) is unsaturated and has a fixed length while the side chains of menaquinones (Vitamin K2) are unsaturated and variable in length. The vitamins are named according to the length of the side chains. VitaminK1 occurs in lettuce, broccoli, cabbage, spinach and vegetable oils while VitaminK2 is found in fermented foods like Natto and various natural cheeses.
Vitamin K2-7 (Menaquinone 7), the physiologically preferred Vitamin K, has a side chain of 7 prenyl residues. It remains biologically active in the body for a longer time than K1. It has been noted that Vitamin K1has half-life of 1.5 hours, whereas Vitamin K2-7 has half-life of 72 hours. Vitamin K2-4 has a half-life of 2 to 3 hours and is used at 1000 times the dose level (in mg) as compared to that for Vitamin K2-7 dose level (in µg). VitaminK2-7therefore is more promising therapeutic choice in enhancing human health.
Calcium plays a critical role in governing human conditions and diseases such as obesity, blood pressure, cardiovascular disease, diabetes etc. The presence of Vitamin K2, calcium and Vitamin D3 in right amounts ensures optimal absorption and binding of calcium into the bone matrix. While Vitamin D3increases the uptake of calcium from the intestine, Vitamin K2 (Vitamin K2-4 andVitamin K2-7) helps incorporate calcium into the bone structure. Vitamin K2 is reported to promote bone formation and suppress bone resorption. While the presence of calcium is necessary for bones, its presence can also lead to calcification of the vessels. In this context Vitamin K2 plays an important role in ensuring that calcium is available in the right places and is not deposited in arteries where its presence could lead to undesirable consequences. By ensuring optimal absorption of calcium in bones, Vitamin K2 helps in protection from osteoporosis, avoid cardiovascular blockages of soft tissue calcification. It also reportedly mediates anti-inflammatory effect during arthritis.
The recommended dosages of Vitamin K1 are ~ 120 µg / day for men and ~ 90 µg / day for women. While no authoritative dosage criteria has been laid down for Vitamin K2-7 it’s dose is recommended based on the Rotterdam study results of 50 % reduction of cardiovascular events, at 45 µg K2-7per day to 360 mcg per day based on the near full carboxylation of Osteocalcin in human physiology. Low levels of Vitamin K2-7 are known to lead to an increased incidence of osteoporosis. It is also known to maintain arterial elasticity. Higher dosages of Vitamin K2 are not reported to cause any adverse effects.
Natto, which is an excellent source of Vitamin K2, is a fermented soy product originated in Japan almost 1000 years ago. However it has not been universally accepted as a source of Vitamin K2 in view of its texture, and flavour. Similarly while cheese is a rich source of Vitamin K2, almost 100g Cheese needs to be consumed to meet the daily minimum requirements of the Vitamin K2. This may not be desirable in view of the high calorific content and the presence of saturated fats, not desirable in cardiovascular conditions.
Vitamin K2-4 and Vitamin K2-7 have been isolated from the natural sources and marketed in various forms. Alternatively synthetic methods have been developed for the synthesis of Vitamin K2-7 as described in US Patents 4320065, 4374775 and 4374290.
Commercially, Vitamin K2-7 products marketed to formulators, are based on Vitamin K2-7 obtained from natural sources and synthetic methods. Vitamin K2-7 is currently marketed in various forms such as oil containing 1500 ppm Vitamin K2-7, cold water dispersible powder containing 2000ppm Vitamin K2-7, Vitamin K2-7 crystals, as Vitamin K2-7 liquid containing 1000 ppm, 2000 ppm or 10000 ppm Vitamin K2-7, soluble in water and granules containing 1500 and 2500 ppm Vitamin K2-7 .The oils used are vegetable oils such as Coconut oil and Sunflower oil and the substrates for powders are microcrystalline cellulose, glyceryl monostearate etc. Several companies market Menaquinone-7, as a dietary source of Vitamin K2-7.Vitamin K2-4 is also available in similar forms.
The formulators then formulate Vitamin K2-7 into various finished products such as tablets, capsules and sachets for consumers using conventional manufacturing techniques such as dry granulation, wet granulation using non-aqueous granulation depending on the form of Vitamin K2-7, roller compaction, film coating and / or combination thereof.
It has been known that Vitamin K is beset with stability issues. It is susceptible to degradation in the presence of light, atmospheric oxygen, ionising radiation, and especially alkaline pH. The stability is especially critical since it affects product quality, safety, dosage and label requirements over the product shelf life. The bulk products are appropriately packed with instructions for storage and handling conditions and shelf life mentioned on it so that the product delivers the anticipated performance over the recommended product life. Additionally, Vitamin K2 also undergoes degradation during processing when the final dosage forms such as tablets, especially when Calcium carbonate based formulations are being manufactured. This precludes the use of aqueous wet granulation technique using Isopropyl alcohol as non-aqueous vehicle, Pearlitol 160C as major diluent and Sodium benzoate as soluble lubricant. Wet granulation in this case is particularly desirable since the quantity of Calcium is high. Wet granulation in aqueous medium is always desirable over that in non-aqueous medium since it eliminates the need for the use and recovery of expensive solvents. Degradation of Vitamin K2 also mandates processing in the absence of light, which poses a serious limitation. Further, while all necessary precautions can be taken at the manufacturing stage, the extent of degradation during storage is often unpredictable. To overcome these problems, it is customary to add overage to the product i.e. add an additional quantity over the label. According to one of the suppliers of the product, the overage could be as high as 50%. Clearly it would be desirable to minimize the overage to minimize the product cost, overcome the limitations on the choice of processing techniques and processing conditions as well as choice of excipients in product formulation and eliminate deleterious effects caused, if any, by the products of degradation, by stabilizing Vitamin K2 prior to formulation.
Realizing the need to stabilize Vitamin K2, Kappa bioscience, one of the manufacturers of Vitamin K2, recently introduced the product K2 Vital Delta VitaminK2-7based on “double microencapsulated” beadlet technology. Gum arabic and starch are used for double coating. However no details of the extent of stability achieved and the advantage accrued have been disclosed. Besides, the double microencapsulation undoubtedly would add to the process steps and product cost. Hence there is a need to develop newer methods of stabilization and formulations of Vitamin K2 which will overcome the processing limitations and enable marketing Vitamin K2 in various product forms, especially hitherto not possible.

Objectives of the invention:
The objectives of the present invention works towards overcoming the shortcomings associated with the Vitamin K compositions seen in the prior art as described below.
In one aspect, the present invention provides compositions comprising Vitamin K and pharmaceutically acceptable excipients which stabilize Vitamin K against the degrading effects of light, moisture and alkalinity, during processing and during storage.
In another aspect, the invention provides compositions comprising Vitamin K and pharmaceutically acceptable excipients which obviate the need for double microencapsulation process for the stabilization of Vitamin K.
In yet another aspect, the invention provides compositions of Vitamin K and pharmaceutically acceptable excipients which enable formulating the compositions in various dosage forms especially tablets, capsules, sachets and emulsions.
The present invention also provides the use of aqueous wet granulation technique in the formulation of Vitamin K based products which are stable to light, moisture and alkalinity.
Another objective of the present invention is to enable the manufacture of formulated Vitamin K2 products in the day light.

Summary of the invention:
It has been surprisingly found that loading Vitamin K onto certain pharmaceutical excipients in particulate form prior to formulating various dosage forms such a tablets , capsules , sachets and emulsions avoids the degradation of Vitamin K in the presence of light, moisture, alkalinity and also obviates the need for double encapsulation as reported in the prior art. Accordingly the present invention relates to compositions comprising Vitamin K and pharmaceutically acceptable excipients selected from sugar pearls, mannitol, dextrose, sorbitol, xylose, crosscaramellose sodium, hydroxypropyl methyl cellulose, ethyl cellulose gelatine , dextran, guar gum, pectin , albumin, chitosan, and carboxymethyl chitosan , which stabilize Vitamin K against light and alkaline pH, obviating the need for microencapsulation or double microencapsulation and enable preparation of a wide range of dosage forms such as tablets, capsules, sachets and emulsions which are stable against light, moisture and alkaline conditions.

Detailed description of the invention:
The present invention involves loading Vitamin K on to pharmaceutically acceptable excipients in the form of microparticles such that the loading of Vitamin K is in the range 10 ppm to 10000 ppm, mixing the loaded microparticles with pharmaceutical excipients using pharmaceutical processing techniques known in the art, under conditions which cannot be used while formulating conventional Vitamin K products. The loading of Vitamin K on to pharmaceutically acceptable excipients in the form of microparticles, helps develop new product forms such as syrups hitherto not possible.
Commercially, Vitamin K2-7 products, marketed to formulators, are based on Vitamin K2-7 obtained from natural sources and synthetic methods. Vitamin K2-7 is currently marketed in various forms such as oil containing 1500 ppm Vitamin K2-7, cold water dispersible powder containing 2000 ppm Vitamin K2-7, Vitamin K2-7 crystals, as Vitamin K2-7 liquid containing 1000 ppm, 2000 ppm or 10000 ppm Vitamin K2-7, soluble in water and granules containing 1500 and 2500 ppm Vitamin K2-7 .The oils used are vegetable oils such as Coconut oil, MCT oil and Sunflower oil and the substrates for powders are microcrystalline cellulose, glyceryl monostearate etc. Several companies market Menaquinone-7, as a dietary source of Vitamin K2-7.
The invention is illustrated below by describing the 1) preparation of Vitamin K2 loaded particles of pharmaceutically acceptable excipients, using aqueous and non-aqueous techniques and appropriate forms of Vitamin K2) preparation of a wide range of products such as tablets, capsules, sachets and emulsions incorporating Vitamin K2 loaded particles of step 1 and 3) evaluation of stability of the products obtained in step 2.

It may be however noted that the examples cited herein are for the sake of illustration only and do not limit the scope of the invention to the cited examples.

Step 1: Loading of Vitamin K2 on to pharmaceutically acceptable carrier particles.
Loading using non aqueous solvents process
A typical procedure for loading Vitamin K2 on sugar microparticles is described below.
Isopropyl alcohol is weighed in a Stainless steel container and kept stirring. Excipient grade Hydroxy propyl methyl cellulose (HPMC) powder is added to the container to obtain a fine slurry, which then turns into a clear solution as stirring is continued. Stock containing Vitamin K2-7 in vegetable oilis then added to this solution. This is followed by the addition of dichloromethane. The clear solution is then filtered through 60 mesh sieve .It is then sprayed on sugar microparticles in a fluid bed processor (Wurster type, bottom spray) to obtain Vitamin K2-7 coated sugar microparticles.Spraying is done at a flow at of 5-125 ml/minute
Typical compositions for loading Vitamin K2 are given below.

Example 1
Sr. No. Ingredients Quantity ( in gms)
1 Sugar Spheres (60-80 Mesh) 500.00
2 VitaminK2-7Oil ** 25.00
3 HPMC 6 cps 50.00
4 Iso-propyl alcohol* 394.00
5 Dichloromethane* 731.00
Total 575.00

*Solvents used in processing do not form a part of the product.
** Vitamin K2-7Oil used is obtained by diluting vitamin K2-7oil obtained by fermentation. The same oil in which the stock is prepared is used for dilution.
Example 2.
Sr. No. Ingredients Quantity ( in gms)
1 Mannitol Granules 500.00
2 Vitamin K2-7 Oil** 25.00
3 HPMC 6 cps 100.00
4 Iso-propyl alcohol* 394.00
5 Dichloromethane* 731.00
Total 625.00

*Solvents used in processing do not form a part of the product.
** Vitamin K2-7 Oil used is obtained by diluting vitamin K2-7 oil obtained by fermentation. The same oil in which the stock is prepared is used for dilution.
Example 3.
Sr. No. Ingredients Quantity
( in gms)
1 Sorbitol Granules 500.00
2 Vitamin K2-7Oil** 25.00
3 HPMC 6 cps 100.00
4 Iso-propyl alcohol* 394.00
5 Dichloromethane* 731.00
Total 625.00

*Solvents used in processing do not form a part of the product..
** Vitamin K2-7 Oil used is obtained by diluting vitamin K2-7 oil obtained by fermentation. The same oil in which the stock is prepared is used for dilution.
Example 4.
Sr. No. Ingredients Quantity
( in gms)
1 Dextrose Granules 500.00
2 Vitamin K2-7Oil** 25.00
3 HPMC 6 cps 100.00
4 Iso-propyl alcohol* 394.00
5 Dichloromethane* 731.00
Total 625.00

*Solvents used in processing do not form a part of the product.
** Vitamin K2-7 Oil used is obtained by diluting vitamin K2-7 oil obtained by fermentation. The same oil in which the stock is prepared is used for dilution.
Example 5.
Sr. No. Ingredients Quantity
( in gms)
1 Xylose Granules 500.00
2 Vitamin K2-7 Oil** 25.00
3 HPMC 6 cps 100.00
4 Iso-propyl alcohol* 394.00
5 Dichloromethane* 731.00
Total 625.00

*Solvents used in processing do not form a part of the product.
** Vitamin K2-7 Oil used is obtained by diluting vitamin K2-7 oil obtained by fermentation. The same oil in which the stock is prepared is used for dilution.
Example 6.
Sr. No. Ingredients Quantity
( in gms)
1 Crosscaramellose sodium Granules 500.00
2 Vitamin K2-7Oil** 25.00
3 HPMC 6 cps 100.00
4 Iso-propyl alcohol* 394.00
5 Dichloromethane* 731.00
Total 625.00

*Solvents used in processing do not form a part of the product.
** Vitamin K2-7 Oil used is obtained by diluting vitamin K2-7 oil obtained by fermentation. The same oil in which the stock is prepared is used for dilution.
Example 7.
Sr. No. Ingredients Quantity
( in gms)
1 Dextran 500.00
2 Vitamin K2-7Oil** 25.00
3 HPMC 6 cps 100.00
4 Iso-propyl alcohol* 394.00
5 Dichloromethane* 731.00
Total 625.00

*Solvents used in processing do not form a part of the product.
** Vitamin K2-7 Oil used is obtained by diluting vitamin K2-7 oil obtained by fermentation. The same oil in which the stock is prepared is used for dilution.
Example 8.
Sr. No. Ingredients Quantity
( in gms)
1 Guar gum 500.00
2 Vitamin K2 Oil** 25.00
3 HPMC 6 cps 100.00
4 Iso-propyl alcohol* 394.00
5 Dichloromethane* 731.00
Total 625.00

*Solvents used in processing do not form a part of the product.
** Vitamin K2-7 Oil used is obtained by diluting vitamin K2-7 oil obtained by fermentation. The same oil in which the stock is prepared is used for dilution.
Example 9.
Sr. No. Ingredients Quantity
( in gms)
1 Pectin 500.00
2 Vitamin K2-7 Oil** 25.00
3 HPMC 6 cps 100.00
4 Iso-propyl alcohol* 394.00
5 Dichloromethane* 731.00
Total 625.00

*Solvents used in processing do not form a part of the product..
** Vitamin K2-7 Oil used is obtained by diluting vitamin K2-7 oil obtained by fermentation. The same oil in which the stock is prepared is used for dilution.

Example 10.
Sr. No. Ingredients Quantity
( in gms)
1 Albumin 500.00
2 Vitamin K2-7 Oil** 25.00
3 HPMC 6 cps 100.00
4 Iso-propyl alcohol* 394.00
5 Dichloromethane* 731.00
Total 625.00

*Solvents used in processing do not form a part of the product.
** Vitamin K2-7 Oil used is obtained by diluting vitamin K2-7 oil obtained by fermentation. The same oil in which the stock is prepared is used for dilution.
Example 11.
Sr. No. Ingredients Specification Qty ( in gms)
1 Chitosan USP/NF 500.00
2 Vitamin K2-7 Oil** 25.00
3 HPMC 6 cps USP/NF 100.00
4 Iso-propyl alcohol* IP 394.00
5 Dichloromethane* IP 731.00
Total 625.00

*Solvents used in processing do not form a part of the product.
** Vitamin K2-7 Oil used is obtained by diluting vitamin K2-7 oil obtained by fermentation. The same oil in which the stock is prepared is used for dilution.
Example 12.
Sr. No. Ingredients Quantity
( in gms)
1 Gelatine 500.00
2 Vitamin K2-7 Oil** 25.00
3 HPMC 6 cps 100.00
4 Iso-propyl alcohol* 394.00
5 Dichloromethane* 731.00
Total 625.00

*Solvents used in processing do not form a part of the product.
** Vitamin K2-7 Oil used is obtained by diluting vitamin K2-7 oil obtained by fermentation. The same oil in which the stock is prepared is used for dilution.
Example 13.
Sr. No. Ingredients Qty ( in gms)
1 Sugar Spheres (60-80 mesh) 500.00
2 Vitamin K2-7Oil** 25.00
3 Ethyl cellulose 100.00
4 Iso-propyl alcohol* 394.00
5 Dichloromethane* 731.00
Total 625.00

*Solvents used in processing do not form a part of the product.
** Vitamin K2-7 Oil used is obtained by diluting vitamin K2-7 oil obtained by fermentation. The same oil in which the stock is prepared is used for dilution.
Example 14
Sr. No. Ingredients Qty ( in gms)
1 Sugar Spheres (60-80 #) 500.00
2 Vitamin K2-4 Oil** 25.00
3 HPMC 6 cps 100.00
4 Iso-propyl alcohol* 394.00
5 Dichloromethane* 731.00
Total 625.00

*Solvents used in processing do not form a part of the product..
** Vitamin K2-4 Oil used is obtained by diluting vitamin K2-4 oil obtained by fermentation. The same oil in which the stock is prepared is used for dilution.

Loading using aqueous solvents
Typical composition used for loading Vitamin K2 is given below.
Hot water is weighed in a Stainless steel container and kept stirring. Excipient grade Ethyl cellulose (EC) powder is added to the container to obtain a fine slurry, which then turns into a clear solution as stirring is continued. This then followed by the addition of Hydroxy ethyl cellulose (HEC) and stirring is continued till a clear solution is obtained. A Stock containing Vitamin K2-7 in vegetable oilis then added to this solution. This is then filtered through 60 mesh sieve .It is then sprayed on sugar micro particles in a fluid bed processor (Wurster type, bottom spray) to obtain Vitamin K2-7coated sugar micro particles.Spraying is done at a flow at of 5-125 ml/minute with continuous mixing of the emulsion.

Example 15
Sr. No. Ingredients Quantity
( in gms)
1 Sugar Spheres (60-80 mesh) 500.00
2 Vitamin K2-7 Oil** 25.00
3 Ethyl cellulose 100.00
4 Hydroxy ethyl cellulose 394.00
5 Hot water* 731.00
Total 625.00

*Solvents used in processing do not form a part of the product.
** Vitamin K2-7 Oil used is obtained by diluting vitamin K2-7 oil obtained by fermentation. The same oil in which the stock is prepared is used for dilution.
Example 16
Acetone is weighed in a Stainless steel container and kept stirring. Excipient grade Ethyl cellulose (EC) powder is added to the container to obtain a fine slurry, which then turns into a clear solution as stirring is continued. This then followed by the addition of water and Hydroxy ethyl cellulose phthalate (HEC) and stirring is continued till a clear solution is obtained. A Stock containing Vitamin K2-7 in vegetable oilis then added to this solution. The clear solution is then filtered through 60 mesh sieve .It is then sprayed on sugar microparticles in a fluid bed processor (Wurster type, bottom spray) to obtain Vitamin K2-7 coated sugar microparticles.Spraying is done at a flow at of 5-125 ml/minute. A typical formulation is described below.
Sr. No. Ingredients Quantity
( in gms)
1 Sugar Spheres (60-80 mesh) 500.00
2 Vitamin K2-7 Oil* 25.00
3 Ethyl cellulose 100.00
4 Hydroxy ethyl cellulose phthalate 394.00
5 Acetone : Water (95:5) 731.00
Total 625.00

*Solvents used in processing do not form a part of the product.
** Vitamin K2-7 Oil used is obtained by diluting vitamin K2-7 oil obtained by fermentation. The same oil in which the stock is prepared is used for dilution.
Step 2: Preparation of pharmaceutical excipient composition for preparing
sachets and capsules and tablets

The Vitamin K loaded pharmaceutical excipient carrier particles prepared according to the procedure and formulations described in preceding paragraphs are subsequently mixed in a second pharmaceutical composition. These compositions can be prepared using a) aqueous wet granulation technique b) non-aqueous wet granulation technique c) direct compression technique.
Each of these is briefly described below along with typical examples of compositions prepared using this technique.
Aqueous wet granulation
Lactose, soluble starch and Calcium carbonate are mixed in requisite quantity of water and treated with starch paste. pH at this stage is between 11-13. The mass is passed through granulator and granules are dried. Then Vitamin K 2-7 loaded micro particles selected from those described in the step1 and Vitamin .D3 are added in a mixer and further mixed with Magnesium stearate. The granules are then used as such by packing into sachets, capsules. The granules are further made into tablets using conventional techniques of tablet making viz. compression.
Non-aqueous wet granulation
Calcium carbonate is wetted with isopropyl alcohol and passed through coarse granulator. The granules are dried, mixed with lactose amylum granules. Vitamin K 2-7 loaded micro particles selected from those described in the step1 and Vitamin .D3 are mixed with the above granules. The dried granules are mixed with Magnesium stearate. The granules are then used as such by packing into sachets, capsules. The granules are further made into tablets using conventional techniques of tablet making.
Direct compression
Calcium carbonate is slugged by passing through a roll compacter 2-3 times. The slugged CaCO3 is then mixed with direct compressible grade lactose. Vitamin K 2-7 loaded micro particles selected from those described in the step1 and Vitamin .D3 are mixed with the above in a mixer. The granules are mixed with Magnesium stearate. The granules are further made into tablets using conventional techniques of tablet making.
For the preparation of tablets, the granules obtained from any of the methods described above are then compressed into tablets following standard roller compaction procedures practiced in pharmaceutical processing.
Compositions of typical products obtained by the above methods are summarized below

Example 17
Vitamin K2-7 loaded on to excipients of the microparticles in step 1 by aqueous wet granulation processed and mixed with the composition of the second step made by aqueous wet granulation process and made into tablets.
Composition/Tablet:
CaCO3: 0.635 gms,
Vitamin K2-7 0.0015 gms,
Vitamin D3 Equivalent to 150 IU.
Lactose/ amylum granules: 0.300 gms,
Mg Stearate: 0.010 gm
Example 18
Vitamin K2-7 loaded on to excipients of the microparticles in step 1 by aqueous wet granulation processed and mixed with the composition of the second step made by aqueous wet granulation process and granules made into sachets
Composition:
CaCO3: 0.635 gms,
Vitamin K2-7 0.0003 gms,
Vitamin D3 Equivalent to 150 IU.
Lactose/ amylum granules: 0.300 gms,
Mg Stearate: 0.010 gm

Example 19
Vitamin K2-7 loaded on to excipients of the microparticles in step 1 by aqueous wet granulation processed and mixed with the composition of the second step made by aqueous wet granulation process and granules made into capsules
Composition:
CaCO3: 0.635 gms,
Vitamin K2-7: 0.0030 gms,
Vitamin D3 Equivalent to 150 IU.
Lactose/ amylum granules: 0.300 gms,
Mg Stearate: 0.010 gm
Example 20

Composition:
CaCO3: 0.635 gms,
Vitamin K2-7: 0.0075 gms,
Vitamin D3: Equivalent to 150 IU.
Lactose/ amylum granules: 0.300 gms,
Mg Stearate: 0.010 gm

Preparation of emulsions
Example 21
Sr. no Ingredients Gms/litre
1 Sorbitol solution 100.00
2 Glycerin 100.00
3 Avicel RC 591 22.50
4 Polysorbate 80 1.00
5 Calcium carbonate # 100.00
6 Xanthan gum 3.00
7 Sucrose 400.00
8 Sodium benzoate 2.50
9 Di sodium EDTA 0.50
10 Citric acid monohydrate 4.50
11 Propylene glycol 125.00
12 Vitamin K2 – 7 loaded on micro particles 0.012
13 Vitamin D3 0.0015
14 Peppermint flavour 4.30
15 Titanium dioxide 1.50
16 Purified water Quantity sufficient to 1litre
A typical procedure for the preparation of emulsions is as follows. Weighed amounts of all ingredients except the flavours are added to half the amount of water and the contents are stirred for 15 minutes. The flavour (s) are then added and the contents are stirred for another 15 minutes. The contents are then stored in tightly closed jars until ready for packing.

Example 22
Sr. No. Ingredients Gms./litre
1 Calcium chloride dihydrate 183.75
2 Sorbitol solution 100.00
3 Glycerin 100.00
4 Sucrose 300.00
5 Sodium benzoate 5.00
6 Vitamin D3 0.00075
7 Vitamin K2 7 loaded on micro particles 0.010
8 Strawberry flavour x
9 Acesulfame potassium x
10 Citric acid anhydrous x
11 Orange flavour x
12 Peppermint flavour 5.500
13 Lemon flavour x
14 Sucralose 2.00
15 Sunset yellow x
16 Quinoline yellow x
17 Purified water Quantity sufficient to 1litre

Example 23
Sr. No. Ingredients Gms/litre
1 Calcium aspartate anhydrous 195.000
2 Sorbitol solution 50.000
3 Glycerin 50.000
4 Propylene glycol 50.000
5 Avicel RC 591 22.500
6 Polysorbate 80 1.000
7 Calcium carbonate 62.500
8 Xanthan gum 1.00
9 Sodium benzoate 2.5
10 Disodium edetate 0.5
11 Citric acid monohydrate 1.00
12 Vitamin D3 0.00075
13 Vitamin K2 7 loaded on micro particles 0.010
14 Acesulfame potassium 10.00
15 Orange flavour 5.50
16 Peppermint flavour 4.3
17 Sucralose x
18 Sunset yellow x
19 Polyethylene glycol x
20 Purified water Quantity sufficient to 1litre

Example 24

Sr. No. Ingredients Gms/litre
1 Calcium chloride dihydrate 183.75
2 Sorbitol 100.00
3 Glycerin 100.00
4 Sucrose 300.00
5 Sodium benzoate 1.00
6 Vitamin D3 0.00075
7 Vitamin K2 7 loaded on micro particles 0.010
8 Strawberry flavour x
9 Acesulfame potassium x
10 Citric acid anhydrous x
11 Orange flavour x
12 Peppermint flavour 5.50
13 Lemon flavour x
14 Sucralose 2.000
15 Sunset yellow x
16 Quinoline yellow x
17 Purified water Quantity sufficient to 1litre

Step 3: Stability study of products
The stability study of the tablets is carried out at 1) room temperature 30 +20C and humidity 65 + 5 % and forced degradation at 40 + 20C and humidity 75 + 5 % as follows.
Tablets containing Vitamin K2-7 along with glyceryl mono stearate were used as a control.
Tablets to serve as control were prepared by normal aqueous granulation. CaCO3 with GMS triturated 2500 ppm Vitamin K2-7 (normal grade) used as control. Vitamin K2-7 was added at the lubrication stage along with vitamin D3 of magnesium stearate.
Tablets using stabilised vitamin K2 were with normal aqueous granulation CacO3, with stabilised 2500 ppm Vitamin K2-7 granules was added at the lubrication stage along with vitamin D3 of magnesium stearate.
For capsules- CaCO3 mix was granulated with aqueous granulation technique. Vitamin K2-7 was added at the lubrication stage along with vitamin D3 of magnesium stearate.
The accelerated aging of Vitamin K2-7 was monitored by estimating the amount of residual VitaminK2-7present in each at the end of specified time intervals.
Vitamin K2-7 and VitaminK2-4 are estimated according the procedures described below.

Estimation of Vitamin K2-7
Mobile Phase: 97% HPLC grade Ethanol.
Take 485mL of HPLC grade Absolute Ethanol and add 15-mL of HPLC Grade water, mix well.
Make up volume up-to 500mL with absolute ethanol. Degas by sonication for at-least 15 minute before use.
HPLC Stationary Phase (Column):
Thermo make BDS-Hypersil C-18, Legth-150mm, ID-4.6mm, and Particle size-5µ
Sample Diluent: HPLC Grade MeOH.
Flow Rate: 0.7 mL / minute.
UV Detector: 254nm.
Amount of Sample Iniection: 100 µL.
Extraction Solvents: Distilled Water, Hexane (AR grade).
Standard Preparation:
a) Weigh -100 mg of working standard* on a calibrated analytical balance having least count of 0.1 mg and transfer to volumetric flask.
b) Add 7 mL MeOH and stopper it.
c) Sonicate at room temperature for at least 15 minutes#
d) Cool and make up the volume to 10ml.
e) Centrifuge at 5000 rpm## for 5 min.
f) Decant and filter supernatant through 0,45 micron filter paper.
g) Store prepared standard sample in refrigerator.
h) Prepare fresh standard every day and use within 24 hour.
* Due to high cost of pure K2-7 standard, a certified working standard can be used instead of pure K2-7standard
'Sonication time should be validated depending upon type and efficacy of sonicator.
"Centrifugation time & speed should be validated depending on type and efficacy of centrifuge.
Sample Preparation:
a) Weigh enough sample to give an area equivalent to -100 mg of standard area. Crush (if
required) sample to make a fine powder. If sample is in liquid, viscous blend or oil form then
take as is.
b) Transfer into evaporating dish I volumetric flask. Add 7 mL of MeOH, sonicate at room
temperature for at least 15 minutes#
c) Cool and make up volume to 10 mL
d) Centrifuge at 4000 rpm ## for 10 min.
e) Take 5 ml of the above supernatant methanol solution in a 50 ml separating funnel.
f) Add 2 ml Distilled Water & 10 ml Hexane. Place stopper on separating fun[!el.
g) Shake vigorously to extract Vitamin K2-7 in the hexane layer.
h) Transfer the aqueous -methanol layer to another 50 ml separating funnel and wash with 10
ml Hexane. (Validate no of washes required)
i) Pool Hexane fractions & evaporate to dryness.
j) Reconstitute with 5 ml HPLC grade Methanol and sonicate.
k) Filter through 0.45 micron filter paper.
Injection:
a) Inject 20 to 1 00 µL of standard and sample supernatant depending on sensitivity of the instrument:
b) Run HPLC as per SOP/LEQ-14.
c) Rinse the syringe with HPLC grade methanol between each injection.
# Sonication time should be validated depending upon type and efficacy of sonicator.
#: Centrifugation time & speed should be validated depending on type and efficacy of centrifuge.
Calculation for content of K2-7
Assay (ppm) = (Area of K2-7 Sample Peak*'Weight of Standard K2-7Standard concentration (ppm)* Sample Dilution) / (Amount of Standard Diluents • Area of K2-7 Standard Peak • Weight of Sample)
For µg/tab- (Area of K2-7 Sample Pea* Weight of Standard *K2-7Standard concentration (ppm)*Sample Dilution)/ (Amount of Standard Diluents • Area of K2-7 Standard Peak' No of tablet taken)
Report as average reading of 4 samples
Abbreviation
HPLC: High Performance Liquid Chromatography.
MeOH: Methanol.
The results for a typical tablet are tabulated below

Example 25
Sample Vitk2-7 ppm 1M*
Acc 1M
RT 2M
Acc 2M
RT 3M
Acc 3M
RT 6M
Acc 6M
RT 9M
RT 12M
RT 24M
RT
Control 28.5 12.46 13.4 9.77 9.67 6.68 6.95 5.49 6.31 4.18 4 3.7
Comparative 28.75 28.75 29.2 26.7 30.9 28.2 28.11 27.83 27.25 27.05 27.19 27.39
Experimental 27.9 25.64 25.1 26.7 29 28.7 28.63 27.99 27.25 27.28 27.12 27.36

• M = Month, Acc = Accelerated testing, RT = Room temperature testing
The results in Table above clearly indicate that VitaminK2-7 loaded on sugar pearls of the present invention, which has not been coated again with ethyl cellulose TiO2 based coating solution on the VitaminK2-7 loaded sugar micro particles, is stabilized as effectively as that of the comparative example which contains glyceryl mono stearate triturated vitamin K2-7 which is the normal grade.
The results further illustrate that the results of stability evaluation under accelerated test conditions for six months are comparable to the stability evaluation under room temperature test conditions for 24 months.
The results of stability evaluation of other products prepared according to the embodiments of the present invention are summarized in examples below.
Following are the results of accelerated aging testing carried out at a temperature: 40°C ± 2°C and Humidity: 75% RH ± 5% RH.
The samples tested were capsules prepared by incorporating Vitamin K2-7 loaded on to excipient micro particles using the wet granulation technique and aqueous medium which were then mixed with the excipient formulation of the second step prepared using wet granulation technique and aqueous medium.
Example 26
Time /Months 0 1 2 3 6
VitaminK2-7 µ g 57 51 55 53 56

Example 27
Time /Months 0 1 2 3 6
VitaminK2-7 µ g 17 14 15 13 16

Example 28
Time /Months 0 1 2 3 6
VitaminK2-7 µ g 116 115 116 114 112

Example 29
Time /Months 0 1 2 3 6
VitaminK2-7 µ g 257 251 255 253 252

Following are the results of accelerated aging testing carried out at a temperature: 40°C ± 2°C and Humidity: 75% RH ± 5% RH.
The samples tested were granules prepared by incorporating Vitamin K2-7 loaded on to excipient micro particles using the wet granulation technique and aqueous medium which were the mixed with the excipient formulation of the second step prepared using wet granulation technique and aqueous medium.

Example 30
Time /Months 0 1 2 3 6
VitaminK2-7 µ g 57 51 55 53 58

Example 31
Time /Months 0 1 2 3 6
VitaminK2-7 µ g 17 14 15 13 16

Example 32
Time /Months 0 1 2 3 6
VitaminK2-7 µ g 116 115 116 114 112

Example 33
Time /Months 0 1 2 3 6
VitaminK2-7 µ g 257 251 255 253 252

Following are the results of accelerated aging testing carried out at a temperature: 40°C ± 2°C and Humidity: 75% RH ± 5% RH.
The samples tested were tablets prepared by incorporating Vitamin K2-7 loaded on to excipient micro particles using the wet granulation technique and aqueous medium which were the mixed with the excipient formulation of the second step prepared using wet granulation technique and aqueous medium.
Example 34
Time /Months 0 1 2 3 6
VitaminK2-7 µ g 57 51 55 53 58
Example 35
Time /Months 0 1 2 3 6
VitaminK2-7 µ g 17 14 15 13 16
Example 36
Time /Months 0 1 2 3 6
VitaminK2-7 µ g 116 115 116 114 112
Example 37
Time /Months 0 1 2 3 6
VitaminK2-7 µ g 257 251 255 253 252

Following are the results of accelerated aging testing carried out at a temperature: 40°C ± 2°C and Humidity: 75% RH ± 5% RH and room temperature aging testing carried out at a temperature: 30°C ± 2°C and Humidity: 65% RH ± 5% RH.
The samples tested were tablets prepared by incorporating Vitamin K2-7loaded on to excipient micro particles using the wet granulation technique and non -aqueous medium which were the mixed with the excipient formulation of the second step prepared using wet granulation technique and non-aqueous medium
Example 38
1M
Acc 1M
RT 2M
Acc 2M
RT 3M
Acc 3M
RT 6M
Acc 6M
RT 9M
RT 12M
RT
Tablets: Vitamin K2-7loaded Particles 54 54 52 53 55 56 52 51 54 52
Tablets : Control 52 48 50 46 48 44 46 48 45 44

Following are the results of accelerated aging testing carried out at a temperature: 40°C ± 2°C and Humidity: 75% RH ± 5% RH.
The sample tested was emulsion prepared by incorporating Vitamin K2-7 loaded on to excipient micro particles using the wet granulation technique and aqueous medium.
Example 39
Time /Months 0 1 2 3 6
VitaminK2-7 µ g 64 61 58 59 63

Following are the results of room temperature aging testing carried out at a temperature: 30°C ± 2°C and Humidity: 65% RH ± 5% RH.
The sample tested was emulsions prepared by incorporating Vitamin K2-7 loaded on to excipient micro particles using the wet granulation technique and aqueous medium
Example 40
Time /Months 0 1 2 3 6 10 12
VitaminK2-7 µ g 64 61 58 59 62 63 61

Vitamin K could be loaded on the micro particles by Wurster process, fluid bed process or pan coating process using techniques well known in the art of pharmaceutical process technology.
Vitamin K loaded micro particles are then mixed with another pharmaceutical excipient composition comprising calcium carbonate, soluble starch and lactose and converted in to various dosage forms such as granules, capsules, tablets and syrups using techniques well known in the art of pharmaceutical process technology.
The products are then subjected to accelerated testing according to standard protocols to monitor the degradation of Vitamin K and the results compared with appropriate controls.
The objectives of the present invention are accomplished and the shortcomings associated with the Vitamin K compositions of the prior art are overcome by the present invention as described below in the preferred embodiments.
According to an embodiment, the present invention describes a stabilized Vitamin K composition comprising Vitamin K loaded on a pharmaceutically acceptable micro particles selected from sugar pearls, mannitol, dextrose, sorbitol, xylose, crosscaramellose sodium, hydroxypropyl methyl cellulose, ethyl cellulose gelatine , dextran, guar gum, pectin , albumin, chitosan, and carboxymethyl chitosan , which stabilizes Vitamin K against light, moisture and alkaline pH.
According to another embodiment of the invention the Vitamin K loaded pharmaceutical substrate microparticles are mixed with another pharmaceutical composition comprising Calcium carbonate, lactose and soluble starch.
According to another embodiment of the present invention the loading of Vitamin K in the micro particles is in the range 10 ppm to 10000 ppm
According to another embodiment the size of the microparticles is in the range 20 mesh to 100 mesh.
According to another embodiment of the present invention Vitamin K is loaded on the micro particles by Wurster process
According to another embodiment of the present invention Vitamin K is loaded on the micro particles by fluidized bed coating process.
According to another embodiment of the present invention Vitamin K is loaded the micro particles by pan coating process
According to another embodiment of the present invention at least 90 % of Vitamin K retains its activity when exposed to ambient light in the course of normal production and formulated product exposure on the shelf life.
According to another embodiment of the present invention at least 90% of Vitamin K retains its activity .at the end of 6 months, in an accelerated stability test, when exposed to alkaline pH condition typically seen in aqueous granulated calcium carbonate tablet formulation.
According to another embodiment of the present invention Vitamin K loaded on to microparticles is Vitamin K2
According to another embodiment of the present invention Vitamin K loaded on to microparticles is Vitamin K2-4
According to another embodiment of the present invention Vitamin K loaded on to microparticles is Vitamin K2-7
The foregoing descriptions of specific embodiments of the present invention have been presented for purposes of illustration and description only. They are not intended to be exhaustive or to limit the present invention to the precise forms disclosed, and obviously many modifications and variations are possible in light of the above teaching.
,CLAIMS:Claims
We claim:
1. A Vitamin K composition in solid form comprising Vitamin K loaded onto pharmaceutical substrates in particulate form mixed with a second pharmaceutical excipient composition wherein the said vitamin K composition is stable.
2. The Vitamin K composition in solid form as claimed in claim 1 wherein the Vitamin K is selected from Vitamin K2-4 and Vitamin K2-7.
3. The Vitamin K composition in solid form comprising Vitamin K loaded onto pharmaceutical substrates in particulate form , wherein the said substrate is selected from a group consisting of sugar pearls, mannitol, dextrose, sorbitol, xylose, crosscaramellose sodium, hydroxypropyl methyl cellulose, ethyl cellulose gelatine , dextran, guar gum, pectin , albumin, chitosan, and carboxymethyl chitosan.
4. The Vitamin K composition in solid form comprising Vitamin K loaded onto pharmaceutical substrates in particulate form as claimed in claim 1 wherein the loading is carried out by wet granulation process using a medium which is aqueous or non-aqueous.
5. The Vitamin K composition in solid form comprising Vitamin K loaded onto pharmaceutical substrates in particulate form mixed with a second pharmaceutical excipient composition as claimed in claim 1 wherein the second pharmaceutical excipient comprises one or more excipient selected from the group consisting of calcium carbonate , lactose and soluble starch
6. The Vitamin K composition in solid form comprising Vitamin K loaded onto pharmaceutical substrates in particulate form mixed with a second pharmaceutical excipient composition as claimed in claim 1, wherein the second pharmaceutical excipient composition is prepared by a dry granulation process.
7. The Vitamin K composition in solid form comprising Vitamin K loaded onto pharmaceutical substrates in the particulate form mixed with a second pharmaceutical excipient composition as claimed in claim 1, wherein the second pharmaceutical excipient composition is prepared by a wet granulation process using non-aqueous or aqueous medium.
8. The Vitamin K composition in solid form comprising Vitamin K loaded onto pharmaceutical substrates in particulate form mixed with a second pharmaceutical excipient composition as claimed in claim 1 , wherein the second pharmaceutical excipient composition is prepared by a roller compaction process.
9. The Vitamin K composition in solid form as claimed in claim 1, wherein the said vitamin K composition is stable during manufacture in the presence of: a) light, b) moisture and c) alkaline pH
10. The Vitamin K composition in solid form as in claim 1, wherein the said composition is stable upon exposure to light, moisture and alkaline pH in storage conditions for at least six months under accelerated test conditions.
11. The Vitamin K composition in solid form as in claim 1, wherein the said composition is stable upon exposure to light, moisture and alkaline pH under storage conditions for at least two years at room temperature .
12. A Vitamin K composition in aqueous liquid form comprising Vitamin K loaded onto pharmaceutical substrates in particulate form mixed with a second pharmaceutical excipient composition in aqueous liquid form wherein the said vitamin K composition is stable.
13. The Vitamin K composition in aqueous liquid form as in claim 12 wherein the Vitamin K is selected from Vitamin K2-4 and Vitamin K2-7.
14. The Vitamin K composition in aqueous liquid form comprising Vitamin K loaded onto pharmaceutical substrates in particulate form as claimed in claim 12 , wherein the said substrate is selected from sugar pearls, mannitol, dextrose, sorbitol, xylose, crosscaramellose sodium, hydroxypropyl methyl cellulose, ethyl cellulose gelatine , dextran, guar gum, pectin , albumin, chitosan, and carboxymethyl chitosan.
15. The Vitamin K composition in aqueous liquid form comprising Vitamin K loaded onto pharmaceutical substrates in particulate form mixed with a second pharmaceutical excipient composition in aqueous liquid form as in claim 12, wherein the second pharmaceutical excipient composition comprises one or more excipient selected from the group consisting of sorbitol , glycerin, calcium carbonate, sucrose, calcium chloride dihydarate, calcium aspartate anhydrous, and propylene glycol,
16. The stabilized Vitamin K composition in aqueous liquid form comprising Vitamin K loaded onto pharmaceutical substrates in particulate form as in claim 12 wherein loading is carried out using a wet granulation process and a medium which is aqueous or non-aqueous.
17. The Vitamin K composition in aqueous liquid form comprising Vitamin K loaded onto pharmaceutical substrates in particulate form mixed with a second pharmaceutical excipient composition as claimed in claim 12, wherein the second pharmaceutical excipient composition comprises one or more excipient selected from the group consisting of calcium carbonate , lactose and soluble starch.
18. The Vitamin K composition in an aqueous liquid form as in claim 12, wherein the said vitamin K composition is stable during manufacture in the presence of: a) light, and b) alkaline pH.
19. The stabilized Vitamin K composition in an aqueous liquid form as claimed in claim 12, wherein the said composition is stable upon exposure to light and alkaline pH in storage conditions for at least six months under accelerated test conditions.
20. The Vitamin K composition in an aqueous liquid form as in claim 12, wherein the said composition is stable upon exposure to light and alkaline pH under storage conditions for at least two years at room temperature test conditions.