The invention relates to food compositions comprising vitamin K and a saturated fat, and in particular to dairy compositions supplemented with vitamin K. The invention further relates to a method for preparing the composition. In addition, the present invention relates to the use of the food composition to increase the bioavailability and activity of vitamin K.

Introduction

Vitamin K is a group of structurally similar, fat-soluble vitamins that are needed for the posttranslational modification of certain proteins, is required for blood coagulation and in metabolic pathways in bone and other tissue. They are 2-methyl-l,4-naphthoquinone (3-) derivatives. This group of vitamins includes two natural vitamins: vitamin Ki and vitamin K2.

Vitamin Ki is also known as vitamin Kj, phylloquinone or

phytomenadione (also called phytonadione). Vitamin Ki is required for coagulation, is synthesized by plants, is found in green leafy vegetables, and can be found in soybean oil.

Vitamin K2 is involved in bone metabolism. Vitamin K2 homologs (menaquinones) are characterized by the number of isoprenoid residues comprising the side chain. Menaquinones are abbreviated MK-n, where n represents the number of isoprenoid side chains. Thus, menaquinone-4 abbreviated MK-4, has 4 isoprene residues in the side chain.

Three synthetic types of vitamin K are known: vitamins K3, K4, and K5. Although the natural Ki and K2 forms are nontoxic, the synthetic form K3 (menadione) has shown toxicity.

Functional foods are an emerging field in food science due to their increasing popularity with health-conscious consumers. Functional food is any food or food ingredient that may provide a health benefit beyond the traditional nutrients comprised in it. The general category of functional foods includes processed food or foods fortified with health -promoting additives, e.g., vitamin enriched products.

An advantage of a functional food over a traditional food supplement containing an active ingredient or ingredients is that the active ingredients are incorporated into a conventional food product so that no separate

administration, e.g. in the form of vitamin supplement tablets, is needed.

Although many foods fortified with food supplements are known, there is a growing need for functional foods providing specific health benefits, e.g. cardiovascular health.

However, a conventional food product supplemented with vitamins and/or minerals does not always exhibit the health benefits expected based on the micronutrients added. The micronutrients added should be compatible with each other and with other components contained in the food product. Moreover, the addition of vitamins may also have an adverse influence on the sensoric characteristics of the supplemented product, such as taste, smell, or consistency. Moreover, good keeping properties and heat-stability of the resulting product are often desired.

Recent research results demonstrate that the small intestine and large intestine (colon) seem to be inefficient at absorbing vitamins K. These results are reinforced by human cohort studies, where a majority of the subjects showed inadequate vitamins K amounts in the body. This was revealed by the presence of large amounts of incomplete gamma-carboxylated proteins in the blood, an indirect test for vitamin K deficiency. So far, vitamin K is often supplemented in an oil dispersion, wherein the oil is a liquid vegetable oil. The vegetable oil is thought to strengthen the intestinal uptake of vitamin K into the blood in order to become bioactive in the mammal. Bruge et al (Br J Nutr 106 (2011) pages 1-5) conducted a study with olive oil supplemented MK-7 in a daily dose of 45 and 90 microgram MK-7. They found that only the 90 microgram dose was able to produce a biological effect. Similar data were found by Theuwissen et al (Br J Nutr 107 (2012) pages 1-6). They studied intake of MK-7 of 0, 10, 20, 45, 90, 180, and 360 microgram per day in an oil composition of natto oil and linseed oil.

Theuwissen et al found only at the high dose (90, 180, 360 microgram/day) a significant effect on the carboxylation of osteocalcin.

Both studies saw that only high doses of vitamin K (90 microgram and higher per day) showed any significant effect on the carboxylation of osteocalcin and/or Matrix Gla Protein (MGP).Van Summeren et al (Br J Nutr 102 (2009) 1171-1178) saw an effect of MK-7 on the osteocalcin carboxylation in prepubertal children with a dose of 45 microgram per day. It should be born in mind that such a dose is the equivalent of 150 microgram per day for adults (0.22 micromolar).

It would desirable to find a matrix wherein lower doses of vitamin K may used that is still as effective as the higher doses of vitamin K as described in the prior art. It is therefor an object of the invention to provide a

composition that is enriched in vitamin K. Another object of the invention is to enhance the bioavailability of vitamin K. Yet another object of the invention is to increase the bioactivity of vitamin K. Furthermore another object of the invention is to maintain, and/or support and/or improve vascular health, especially vascular flexibility and/or blood flow. Another object of the invention is to prevent and/or stabilise metabolic syndrome, and/or diabetes. Yet another object of the invention is to maintain and/or support and/or improve mental health.

It has been found by the present inventors that the bioavailability and stability of vitamin K in blood depends on the type of fat used and the type of vitamin K. In particular, vitamin K2, and especially MK-7, in combination with a fat having at least 36 wt% saturated fatty acids, such as e.g. animal fat, and especially butterfat, or and/or milk fat results in a higher concentration of vitamin K in blood than when administered in combination with more unsaturated liquid oils, such as sunflower oil, olive oil and/or linseed oil.

Summary of the invention

In a first aspect the invention relates to a food composition comprising fat and vitamin K2, wherein the fat comprises at least 36% saturated fatty acids, and wherein the vitamin K2 is in the form MK-7 in an amount of at least 0.1 meg MK-7 per daily intake. In a preferred embodiment of the present invention, the fat is selected from the group comprising palm oil, palm olein, palm kernel oil, shea nut, illipe, cottonseed oil, coconut oil, cocoa butter, butterfat, milk fat, lard, suet, tallow, fractions thereof, and any combination thereof, and preferably is animal fat. In a preferred embodiment the fat is butterfat, butter oil, anhydrous milk fat (AMF), milk fat and/or fractions thereof.

In a preferred embodiment of the present invention and/or embodiments thereof, the composition comprises further milk or milk derived ingredients. Preferably, the composition of the present invention and/or embodiments thereof comprise a milk derived ingredient selected from the group consisting of casein, lactalbumin, lactoglobulin, hydrolyzed casein, hydrolyzed whey proteins, casemates, lactose, curd, milk powder, milk solids, butter solids, butter oil, lactic acid, and/or recaldent. It should be clear that combinations of milk derived ingredients are envisioned in the present invention and/or embodiments thereof.

In another preferred embodiment of the present invention and/or embodiments thereof the composition comprises 0.20-400 meg vitamin MK-7 per daily intake. More preferably, 0.5-400 meg per daily intake or per serving. Suitably the composition according to the invention and/or embodiments thereof comprise MK-7 in the range of 1-350 meg per daily intake, more suitably from 2-300 meg per daily intake, more preferably from 5-250 meg MK- 7 per daily intake, more preferably from 10 -200 meg MK-7 per daily intake, more preferably from 15- 180 meg MK-7 per daily intake, even more preferably from 20-150 meg MK-7 per daily intake, even more preferably from 25-120 meg MK-7 per daily intake, more preferably from 40-100 meg MK-7 per daily intake and most preferably from 50-90 meg MK-7 per daily intake. Since vitamin K promotes blood clotting, which can be undesired, the upper limit for MK-7 in the composition is preferably less than 400 meg MK-7 per daily intake or per serving, and more preferably less than 200 meg MK-7 and most preferably less than 100 meg MK-7 per daily intake.

In another preferred embodiment of the present invention and/or embodiments thereof the composition further comprises an ingredient selected from the group comprising calcium, magnesium, potassium, polyunsaturated fatty acids and vitamin D. It is to be understood that combinations of these ingredients are possible.

In a second aspect, the invention is directed to a method of preparation of the composition according to the invention and/or embodiments thereof, comprising adding vitamin MK-7 to a composition comprising a fat comprising at least 36 wt% saturated fatty acids. Optionally a premix of vitamin MK-7 is added to the food product. The premix, defined as something that is mixed or blended from two or more ingredients or elements before being used or mixed further during product preparation, may suitably comprise other vitamins and minerals. Suitably the MK-7 is first mixed to a fruit preparation and/or fruit syrup and then added to a food product comprising a fat with at least 36 wt% saturated fatty acids.

Another aspect of the invention relates to a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve vascular health, in particular cardiovascular health. The invention further relates to a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve vessel flexibility.

The invention further relates to a composition according to the invention and/or embodiments thereof to prevent and/or maintain and/or reduce arterial stiffness.

The invention further relates to a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve bone health.

The invention further relates to a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve normal clotting capability and/or to maintain normal platelet aggregation and/or to prevent adherence of platelets to blood vessel walls and/or fatty plaques.

The invention further relates to a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve glucose control and/or insulin secretion and/or insulin sensitivity.

The invention further relates to a composition according to the invention and/or embodiments thereof to prevent and/or maintain and/or reduce chronic inflammation and/or low grade inflammation.

The invention further relates to a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve mental health and/or cognitive performance and/or alertness, and/or mental sharpness and/or memory.

The invention further relates to a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve a healthy bodyweight, and/or a healthy body mass index (BMI), and/or a healthy triglyceride blood levels, and/or a healthy waist circumference. The health status of all health parameters mentioned in this invention is

preferably according to the World Health Organization guidelines (WHO guidelines for the prevention, management and care of diabetes mellitus, 2006; Global strategy on diet, physical activity and health, WHO report 2004).

The invention further relates to a composition according to the invention and/or embodiments thereof to maintain a healthy blood pressure and/or to prevent and/or reduce high blood pressure (according to WHO guidelines).

The invention further relates to a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve blood flow.

The invention further relates to a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve blood circulation, including the macrocirculation and/or the

microcirculation .

The invention further relates to a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve blood perfusion. This is the process of delivery of blood to a capillary bed to tissues of a mammal, such as to the brain, kidney, heart, lung, liver, limbs and to the gastrointestinal tract tissue.

The invention further relates to a composition according to the invention and/or embodiments thereof to prevent and/or maintain and/or reduce metabolic syndrome.

The invention further relates to a composition according to the invention and/or embodiments thereof for use in the treatment of vascular disease, and/or cardiovascular disease, and/or atherosclerosis, and/or arterial stiffness and/or hypertension and/or clogged arteries and/or varicose veins and/or transient ischemic attack (TIA) and/or stroke and/or cold limbs.

The invention further relates to a composition according to the invention and/or embodiments thereof for use in the treatment of vessel inflexibility. The invention further relates to a composition according to the invention and/or embodiments thereof for use in the treatment of bone demineralisation and/or osteoporosis.

The invention further relates to a composition according to the invention and/or embodiments thereof for use in the treatment of thrombosis and/or stroke.

The invention further relates to a composition according to the invention and/or embodiments thereof for use in the treatment of diabetes and/or insulin resistance.

The invention further relates to a composition according to the invention and/or embodiments thereof for use in the treatment of chronic inflammation and/or low grade inflammation.

The invention further relates to a composition according to the invention and/or embodiments thereof for use in the treatment of mental decline and/or mental disease, such as dementia and/or Alzheimer disease.

The invention further relates to a composition according to the invention and/or embodiments thereof for use in the treatment of obesity, and/or metabolic syndrome, and/or high BMI, and/or high triglyceride in blood, and/or high body weight, and/or high waist circumference.

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve vascular health, and/or cardiovascular health.

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve vessel flexibility

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to prevent and/or maintain and/or reduce arterial stiffness.

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to maintain a healthy blood pressure and/or to prevent and/or reduce high blood pressure (according to WHO guidelines).

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve blood flow.

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve blood circulation, including the macrocirculation and/or the

microcirculation .

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to support, maintain and/or improve blood perfusion. This is the process of delivery of blood to a capillary bed to tissues of a mammal, such as to the brain, kidney, heart, lung, liver, limbs and to the gastrointestinal tract tissue.

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve bone health.

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve normal clotting capability and/or to maintain normal platelet aggregation and/or to prevent adherence of platelets to blood vessel walls and/or fatty plaques.

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve glucose control mechanism, and/or insulin secretion and/or insulin sensitivity, and/or to prevent diabetes and/or hyperglycemia.

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to prevent and/or maintain and/or reduce to chronic inflammation and/or low grade inflammation. The invention further relates to the use of a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve mental health and/or cognitive performance and/or alertness and/or memory and/or mental sharpness.

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve healthy weight management and/or a healthy body mass index and/or a healthy triglyceride blood levels and/or to prevent the increase of body fat mass.

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to support and/or maintain a healthy waist circumference and/or to reduce a high waist circumference.

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to prevent and/or maintain and/or reduce metabolic syndrome.

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to enhance bioavailability of vitamin K and/or vitamin K2 and/or MK-7.

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to increase the biological activity of vitamin K and/or vitamin K2 and/or MK-7.

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to increase the level of vitamin K and/or vitamin K2 and/or MK-7 in blood of a mammal.

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to reduce the level of circulating uncarboxylated MGP and/or uncarboxylated osteocalcin in blood in a mammal.

The invention further relates to the use of a composition according to the invention and/or embodiments thereof to increase the level of circulating carboxylated MGP and/or carboxylated osteocalcin in blood in a mammal. In a preferred embodiment of the present invention and/or

embodiments thereof the reduction of the level of circulating uncarboxylated MGP and/or uncarboxylated osteocalcin, and/or the increase of the level of circulating carboxylated MGP and/or carboxylated osteocalcin in blood in a mammal and/or the increase of the level of vitamin K and/or vitamin K2 and/or MK-7 in blood of a mammal is obtained within 12 weeks, preferably within 8 weeks, more preferably within 4 weeks.

The invention further relates to a method to reduce the level of circulating uncarboxylated MGP and/or uncarboxylated osteocalcin in blood of a mammal by administering to a mammal a composition according to the invention and/or embodiments thereof.

The invention further relates to a method to increase the level of circulating carboxylated MGP and/or carboxylated osteocalcin in blood of a mammal by administering to a mammal a composition according to the invention and/or embodiments thereof.

The invention further relates to a method to increase the level of vitamin K2 in blood of a mammal by administering to a mammal a

composition according to the invention and/or embodiments thereof.

Preferably the composition according to the invention and/or embodiments thereof in a method according to the invention and/or

embodiments thereof is administered for a period of at least 4 weeks, preferably at least 8 weeks and most preferably at least 12 weeks. Preferably the composition of the present invention and/or embodiments thereof is administered life-long, suitably 3, 5, 7, 10, 12, 15, 18, 20, 23, 25, 27, 30, 32, 35, 38, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or even up to 95years. The intake of vitamin K and/or vitamin K2 and/or MK-7 is preferably daily, each day, so as to maintain a steady level of vitamin K, vitamin K2 and/or MK-7 in the blood. The products of the present invention may easily be incorporated in a normal healthy diet, so that each day, a life long, the vitamin K, vitamin K2 and/or MK-7 level is improved. Detailed description

An ingredient of the composition according to the present invention is vitamin K2, or menaquinone. Vitamin K2 is a member of the vitamin K group of which all members share a methylated naphthoquinone ring structure, and vary in the aliphatic side chain attached at the 3-position.

Menaquinones have side chains composed of a variable number of unsaturated isoprenoid residues; generally they are designated as MK-n, where n specifies the number of isoprenoids.

The group of vitamin K is involved in blood clotting, and in calcium metabolism. Vitamin K is commonly found in multiple vitamin supplements and in infant formula. However, it is mostly present in the form of vitamin Kl and therefore a general designation to vitamin K refers in fact to vitamin Kl, since this form is considerably less expensive to obtain and it has been generally recognised as safe in food since a long time. It should be noted that vitamine Kl, phyloquinone, is different from vitamine K2, menaquinone.

The vitamin K2 in the present invention is in the form of MK-7, i.e. menaquinone having side chains composed of 7 unsaturated isoprenoid residues. It is to be understood that when in the present description vitamin K is mentioned, vitamin K2 is meant, and more specifically MK-7. Vitamin K2, such as MK-7 may be obtained from any suitable source, e.g., fermented soy product such as natto. Also synthetically prepared vitamin K2 is suitable having an advantage that the quality of such obtained vitamin K2 can easily be controlled. However, vitamin K2 obtained from a natural source, e.g., by microbial production, may be preferred. Cheese naturally contains vitamin K2 which has longer isoprenoid chains, usually MK-8, MK-9 and longer.

The content of MK-7 and other ingredients in the composition according to the invention is characterised in this description in amounts per daily intake, or in amounts per serving or in amounts per weight or volume product. It is common to express the amount of vitamins in a daily amount. Many national regulations on labelling of food products indicate the amounts of vitamins as recommended daily intake (RDI) or recommended daily allowance (RDA), see e.g. for the US the electronic code of federal regulations title 21 §101 (http://www.ecfr.gov/cgi-bin/text- idx?c=ecfr&sid=8c5344f04a8ael03e5b0ff5al7c7fa97&rgn= div8&view=text&no de=21:2.0.1.1.2.4. l. l&idno=21). In the EU according to Directive 90/496 EEC directed to labelling rules of nutrients, information on vitamins and minerals must be expressed as a percentage of the recommended daily allowance (RDA). For the purpose of the present invention, amounts are indicated in microgram (meg), nanogram (ng), milligram (mg) or gram (g). A serving is about 125 gram for a viscous (spoonable) product, or about 200-250 ml for a pourable product such as milk or a yogurt drink. For cheese, a serving means one slice of about 20-40 g, and for a spread or butter a serving is about 3.5-7 gram normally used with a one slice of bread. Daily intake means the total amount of an ingredient as consumed within the product of products of the invention per day. A daily intake may be consumed in one serving, but can also be divided over 2, 3, 4 or even more servings per day. The daily intake can also be spread over several products, such as milk and yogurt, milk and cheese, etc. A skilled person is able to find an appropriate amount of servings, or an appropriate combination of products, based on the provided information. For example, spoonable and drink yogurts are usually consumed once or twice per day, therefore the daily intake is generally one to two servings per day. For cheese, a daily intake usually means one slice per day, but can also be more. For a spread or butter, a daily intake usually means about 4 servings, assuming that on average 4 slices of bread are consumed per day. However, also lower or higher amounts of servings are possible, depending on the type and content of the product and depending also on consumer's habits.

In the present invention the amount of MK-7 is indicated in micrograms (meg). The composition according to the present invention preferably comprises at least 0.1 meg MK-7 per daily intake or per serving. Preferably, the composition contains at least 0.2 meg MK-7 per daily intake, and more preferably, in the range 0.5-400 meg per daily intake or per serving. Suitably the composition according to the invention and/or embodiments thereof comprise MK-7 in the range of 1-350 meg per daily intake, more suitably from 2-300 meg per daily intake, more preferably from 5-250 meg MK- 7 per daily intake, more preferably from 10 -200 meg MK-7 per daily intake, more preferably from 15-180 meg MK-7 per daily intake, even more preferably from 20-150 meg MK-7 per daily intake, even more preferably from 25-120 meg MK-7 per daily intake, more preferably from 40-100 meg MK-7 per daily intake and most preferably from 50-90 meg MK-7 per daily intake. Since vitamin K promotes blood clotting, which can be undesired, the upper limit for MK-7 in the composition is preferably less than 400 meg MK-7 per daily intake or per serving, and more preferably less than 200 meg MK-7 and most preferably less than 100 meg MK-7 per daily intake.

Suitably, for a spoonable product such as yoghurt, creme fraiche, or a desert, the amount of MK-7 is at least 0.1 mcg/100 gram product. In a preferred embodiment of the present invention and/or embodiments thereof, the amount of MK-7 is from 0.2-400 mcg/100 gram product, more preferably from 0.5-350 meg MK7/100 gram product, more preferably from 1-250 meg MK7 per 100 gram product, more preferably from 5-200 meg MK7 per 100 gram product, more preferably from 10-150 meg MK-7 per 100 gram product, more preferably from 20-100 meg MK-7 per 100 gram product, more preferably from 40-90 meg MK7 per 100 gram product, more preferably from 50-70 meg MK-7 per 100 gram product, even more preferably from 9-90 meg MK7 per 100 gram product. Suitably the amount of MK-7 is less than 200 mcg/100 gram product, preferably less than 100 mcg/100 gram product, preferably less than 50 mcg/100 gram product, preferably less than 40 mcg/100 gram product preferably less than 20 mcg/100 gram product.

Suitably for a drink, such as milk, yoghurt drink, dairy beverage, fruit beverage, the amount of MK-7 is at least 0.1 meg per 100 ml of product. In a preferred embodiment of the present invention and/or embodiments thereof, the amount of MK-7 is from 0.2-400 mcg/100 ml product, more preferably from 0.5-350 meg MK7/100 ml product, more preferably from 1-250 meg MK7 per 100 ml product, more preferably from 5-200 meg MK7 per 100 ml product, more preferably from 10-150 meg MK-7 per 100 ml product, more preferably from 20-100 meg MK-7 per 100 ml product, more preferably from 40-90 meg MK7 per 100 ml product, more preferably from 50-70 meg MK-7 per 100 ml product, even more preferably from 6-60 meg MK7 per 100 ml product. Suitably the amount of MK-7 is less than 200 mcg/100 ml product, preferably less than 100 mcg/100 ml product, preferably less than 60 mcg/100 ml product, preferably less than 40 mcg/100 ml product preferably less than 20 mcg/100 ml product.

Suitably for hard cheese that is eaten in slices, the amount of MK-7 is at least 0.25 meg per 100 gram of product. In a preferred embodiment of the present invention and/or embodiments thereof, the amount of MK-7 is from 0.5-1000 mcg/100 gram product, more preferably from 1.5-900 meg MK7/100 gram product, more preferably from 2.5-750 meg MK7 per 100 gram product, more preferably from 10-500 meg MK7 per 100 gram product, more preferably from 25-400 meg MK-7 per 100 gram product, more preferably from 50-250 meg MK-7 per 100 gram product, more preferably from 100-200 meg MK7 per 100 gram product, more preferably from 120-170 meg MK-7 per 100 gram product, even more preferably from 45-450 meg MK7 per 100 gram product. Suitably the amount of MK-7 is less than 400 mcg/100 gram product, preferably less than 200 mcg/100 gram product, preferably less than 100 mcg/100 gram product, preferably less than 60 mcg/100 gram product preferably less than 40 mcg/100 gram product.

Suitably for a spread such as butter, margarine, dairy spread, cheese spread, the amount of MK-7 is at least 0.2 meg per 100 gram of product. In a preferred embodiment of the present invention and/or embodiments thereof, the amount of MK-7 is from 0.5-1000 mcg/100 gram product, more preferably from 1.5-900 meg MK7/100 gram product, more preferably from 2.5-750 meg MK7 per 100 gram product, more preferably from 10-500 meg MK7 per 100 gram product, more preferably from 25-400 meg MK-7 per 100 gram product, more preferably from 50-250 meg MK-7 per 100 gram product, more preferably from 100-200 meg MK7 per 100 gram product, more

preferably from 120-170 meg MK-7 per 100 gram product, even more preferably from 90-900 meg MK7 per 100 gram product. Suitably the amount of MK-7 is less than 500 meg/ 100 gram product, preferably less than 400 mcg/100 gram product, preferably less than 200 mcg/100 gram product, preferably less than 100 mcg/100 gram product preferably less than 50 mcg/100 gram product, preferably less than 30 meg MK-7 per 100 gram product.

Good results are obtained when the food composition comprises 20- 60 meg of MK-7 per daily intake or per serving (on top of there daily intake of vitamin K2 from other food products). It is to be understood that in the western world, the normal daily intake of vitamin K2, is about 30 meg from an average diet, which is not sufficient. Vitamin Ki is found chiefly in leafy green vegetables such as spinach, swiss chard, and Brassica (e.g. cabbage, kale, cauliflower, broccoli, and brussels sprouts); some fruits such as avocado, kiwifruit and grapes are also high in vitamin K. Colonic bacteria synthesize a significant portion of humans' vitamin K needs; this is one of the reasons why newborns often receive a vitamin K shot at birth - in order to tide them over until day 5-7 when their colon becomes colonized. Menaquinone-4 (vitamin K2) is synthesized by animal tissues and is found in meat, eggs, and dairy products. Menaquinone-7 is synthesized by bacteria during fermentation and is found in fermented soybeans (natto). The composition according to the invention comprises a fat having at least 36 wt% saturated fatty acids.

In the present invention, the amount of fatty acids is given in percent by weight (wt%) of total fatty acids. In a preferred embodiment of the present invention and/or embodiments thereof the fat comprises between 40 and 95 wt% saturated fatty acids, more preferably between 43 and 90 wt% saturated fatty acids, more preferably between 45 and 80 wt% saturated fatty acids, more preferably between 48 and 70 wt% saturated fatty acids, even more preferably between 52 and 68 wt% saturated fatty acids, even more preferably between 55 and 65 wt% saturated fatty acids, even more preferably between 58 and 63 wt% saturated fatty acids, even more preferably between 58 and 61 wt% saturated fatty acids., The table below shows the common names, structural formula and lipid numbers of saturated fatty acids.

Structural Lipid

Common Name Systematic Name

Formula Numbers

Propionic acid Propanoic acid CH3CH2COOH C3:0

Butyric acid Butanoic acid CH ₃ (CH ₂ )2COOH C4:0

Valeric acid Pentanoic acid CH ₃ (CH ₂ )3COOH C5:0

Caproic acid Hexanoic acid CH ₃ (CH ₂ )4COOH C6:0

Enanthic acid Heptanoic acid CH ₃ (CH ₂ )5)COOH C7:0

Caprylic acid Octanoic acid CH ₃ (CH ₂ ) ₆ COOH C8:0

Pelargonic acid Nonanoic acid CH ₃ (CH ₂ ) ₇ COOH C9:0

Capric acid Decanoic acid CH ₃ (CH ₂ ) ₈ COOH C10:0

Undecylic acid Undecanoic acid CH ₃ (CH ₂ ) ₉ COOH C11:0

Laurie acid Dodecanoic acid CH ₃ (CH ₂ )ioCOOH C12:0

Tridecylic acid Tridecanoic acid CH ₃ (CH ₂ )iiCOOH C13:0

Myristic acid Tetradecanoic acid CH ₃ (CH ₂ )i ₂ COOH C14:0

Pentadecylic acid Pentadecanoic acid CH ₃ (CH ₂ )i ₃ COOH C15:0

Palmitic acid Hexadecanoic acid CH ₃ (CH ₂ )i ₄ COOH C16:0

Margaric acid Heptadecanoic acid CH ₃ (CH ₂ )i ₅ COOH C17:0

Stearic acid Octadecanoic acid CH ₃ (CH ₂ )i ₆ COOH C18:0

Nonadecylic acid Nonadecanoic acid CH ₃ (CH ₂ )i ₇ COOH C19:0

Arachidic acid Eicosanoic acid CH ₃ (CH ₂ )i ₈ COOH C20:0

Heneicosylic acid Heneicosanoic acid CH ₃ (CH ₂ )i ₉ COOH C21:0 Behenic acid Docosanoic acid CH ₃ (CH ₂ )2oCOOH C22: :0

Tricosylic acid Tricosanoic acid CH ₃ (CH ₂ )2iCOOH C23: :0

Lignoceric acid Tetracosanoic acid CH ₃ (CH ₂ )22COOH C24: :0

Pentacosylic acid Pentacosanoic acid CH ₃ (CH ₂ )23COOH C25: :0

Cerotic acid Hexacosanoic acid CH ₃ (CH ₂ )24COOH C26: :0

Heptacosylic acid Heptacosanoic acid CH ₃ (CH ₂ )2 ₅ COOH C27: :0

Montanic acid Octacosanoic acid CH ₃ (CH ₂ )2 ₆ COOH C28: :0

Nonacosylic acid Nonacosanoic acid CH ₃ (CH ₂ )27COOH C29: :0

Melissic acid Triacontanoic acid CH ₃ (CH ₂ )2 ₈ COOH C30: :0

Henatriacontylic Henatriacontanoic

Lacceroic acid Dotriacontanoic acid CH ₃ (CH ₂ ) ₃₀ COOH C32: :0

Psyllic acid Tritriacontanoic acid CH ₃ (CH ₂ ) ₃ iCOOH C33: :0

Tetratriacontanoic

Geddic acid CH ₃ (CH ₂ ) ₃ 2COOH C34: :0 acid

Pentatriacontanoic

Ceroplastic acid CH ₃ (CH ₂ ) ₃₃ COOH C35: :0 acid

Hexatriacontylic Hexatriacontanoic

CH ₃ (CH ₂ ) ₃₄ COOH C36: :0 acid acid

The following table gives the fatty acid composition of some dietary fats.

Saturated Monounsaturated Polyunsaturated

Animal fat

Lard 41 47 12

Duck fat 34 52 14

Butter (Cow) 68 28 4

Butter (Goat) 69 27 4

Beef tallow 52 44 4

Vegetable fat

Coconut oil 92 6 2

Allanblackia 54 46 0

Canola 6 62 32

Palm oil 50 40 10

Palm olein 43 46 11

Palm kernel 83 15 2 Cottonseed oil 25 20 55

Cocoa butter 63 34 3

Soya oil 15 24 61

Illipe 62 37 1

Olive oil 14 75 11

Corn oil 13 28 59

Sunflower oil 12 19 69

Safflower oil 9 13 78

Hemp oil 10 15 75

Shea nut oil 47 46 7

Linseed oil 10 21 69

Most common saturated fatty acids are butyric acid, lauric yristic acid, palmitic acid, and stearic acid.

Saturated fat profile of common foods;

Esterified fatty acids as percentage of total fat

Lauric acid Myristic acid Palmitic acid Stearic acid

Food

C12:0 C14:0 C16:0 C18:0

Coconut oil 47% 18% 9% 3%

Butter 3% 11% 29% 13%

Ground beef 0% 4% 26% 15%

Dark

0% 1% 34% 43% chocolate

Salmon 0% 1% 29% 3%

Eggs 0% 0.3% 27% 10%

Cashews 2% 1% 10% 7%

4%

Soybean oil 0% 0% 11% In a preferred embodiment the saturated fatty acid is selected from the group of lauric acid, myristic acid, palmitic acid, and stearic acid. More preferably the saturated fatty acid is selected from the group of myristic acid and palmitic acid. Suitably the fat of the present invention and/or embodiments thereof comprises more longer saturated fatty acids, such as C14:0, C16:0, and C18:0 than shorter saturated fatty acids such as C4:0, C10:0 and C12:0. In a preferred embodiment of the present invention and/or embodiments thereof the ratio of saturated fatty acids having 1-12 carbon atoms (C1-C12) to saturated fatty acids having more than 12 fatty acids (C 14-30) less than 2, more preferably the ratio of Cl-12 to C14:30 is less than 1.5, more preferably less than 1, even more preferably less than 0.5 and even more preferably less than 0.3, more preferably less than 0.2 and most preferably less than 0.15.

Preferably the fat of the present invention comprises less than 40 wt% short saturated fatty acids having 1-12 carbon atoms (Cl-12), more preferably the fat of the present invention comprises less than 35 wt% Cl-12 saturated fatty acids, even more preferably less than 30 wt% Cl-12 saturated fatty acids, even more preferably less than 25 wt% Cl-12 saturated fatty acids, and more preferably less than 20 wt% Cl-12 saturated fatty acid, most preferably less than 15 wt% Cl-12 saturated fatty acids. The weight

percentages are on total fatty acids.

In another preferred embodiment of the invention and/or embodiments thereof the combined amount of myristic and palmitic acid is more than 13 wt% on total fatty acids. More preferably the combined amount of myristic and palmitic acid is more than 15 wt%, more preferably more than 17 wt%, more preferably more than 20 wt%, more preferably more than 25 wt%, even more preferably more than 27 wt%, and even more preferably more than 30 wt% and most preferably more than 33 wt% on total fatty acids.

In a preferred embodiment the fat is selected from the group consisting of palm oil, palm olein, palm kernel oil, shea nut, Illipe, cottonseed oil, coconut oil, cocoa butter, butterfat, butter oil, anhydrous milk fat, milk fat, lard, suet, tallow, fractions thereof, and any combination thereof, and preferably is an animal fat. Any food-grade animal fat may be used. Animal fat is defined as obtained from an animal source in contrast to a vegetable source and is preferably obtained from mammals, and/or poultry in the commercial processes of rendering or extracting. Preferred animal fats may be butterfat, milk fat, lard, suet, or fractions thereof. Animal fat also includes fat obtained from milk of mammals. In a preferred embodiment the fat is a milk derived fat such milk fat, butter fat, butter oil, anhydrous milk fat (AMF), phospholipids and/or sphingolipid enriched milk fat and fractions thereof. In a particularly preferred embodiment, the animal fat is butterfat, milk fat and/or fractions thereof. Combinations of saturated fat are expressly envisioned. In the present invention and/or embodiments thereof it is also possible to have combinations of one or more fats having at least 36 wt% saturated fatty acids and one or more fats having less than 36 wt% saturated fatty acids, as long as the total amount of saturated fatty acids of the combined fats is at least 36 wt%.

In a preferred embodiment of the present invention and/or embodiments thereof, the fat does not comprise hydrogenated fat. For health reasons, hydrogenated fats are not used, or are kept to a minimum amount. In a preferred embodiment of the present invention and/or embodiments thereof, the amount of hydrogenated fat is less than 10 wt% of the fat phase, more preferably less than 8 wt% of the fat phase, more preferably less than 6 wt% of the fat phase, more preferably less than 4 wt% of the fat phase, even more preferably less than 2 wt% of the fat phase, even more preferably less than 1 wt% of the fat phase, and most preferably the fat phase comprises 0 wt% of hydrogenated fat.

In a preferred embodiment of the present invention and/or embodiments thereof, the fat comprises fish oil or other LC-PUFA (long-chain poly-unsaturated fatty acids) rich oil. It is to be understood that the total amount of saturated fatty acids should not be less than 36 wt%. Preferred amounts of LC-PUFA are 1-1000 mg per 100 gram product. More preferable amount of LC-PUFA are 5-500 mg per 100 gram product, 10-300 mg/100 gram product, more preferably 20-200 mg per 100 gram product, more preferably 25- 150 mg per 100 gram product, more preferably 30-120 mg per 100 gram product and most preferably 40-100 mg per 100 gram product. In another embodiment of the present invention and/or embodiments thereof the amount of LC-PUFA is 5-5000 mg per daily dose, more preferably 10-3000 mg LC- PUFA per daily dose, more preferably 20-2000 mg LC-PUFA per daily dose, more preferably 50-1000 mg LC-PUFA per daily dose, even more preferably 60-500 mg LC-PUFA per daily dose, more preferably 75-200 mg LC-PUFA per daily dose and most preferably 80-160 mg LC-PUFA per daily dose.

LC-PUFAs are fatty acids that contain more than one double bond in the backbone and preferably have at least 20 carbon atoms in the molecule. This class includes many important compounds, such as essential fatty acids, e.g., omega-3 and omega-6 fatty acids and preferably omega-3 fatty acids.

According to the present invention and/or embodiments thereof the LC-PUFA may be selected from the group consisting of cis-6,9, 12- octadecatirenoic acid (GLA, gamma linolenic acid), cis- 11, 14, 17-eicosatrienoic acid (ETE) C20:3, cis-8, ll, 14, 17-eicosatetraenoic acid (ETA) C20:4, cis- 5,8, 11,14, 17-eicosapentaenoic acid (EPA) C20:5, cis-7, 10, 13, 16, 19- docosapentaenoic acid (DPA, Clupanodonic acid) C22:5, cis-4, 7, 10, 13, 16, 19- docosahexaenoic acid (DHA) C22:6, «s-9, 12, 15, 18,21-tetracosapentaenoic acid C24:5; «s-6,9, 12,15, 18,21-tetracosahexaenoic acid (Nisinic acid) C24:6, cis- 11, 14-eicosadienoic acid C20:2, cis-8, 11, 14-eicosatrienoic acid (Dihomo-gamma- linolenic acid) (DGLA) C20:3, «s-5,8, ll, 14-eicosatetraenoic acid (Arachidonic acid) (AA) C20:4, cis- 13, 16-docosadienoic acid C22:2, cis-l, 10, 13, 16- docosatetraenoic acid (Adrenic acid) C22:4, «s-4,7, 10, 13, 16-docosapentaenoic acid (Osbond acid) C22:5. Most preferred are EPA, DHA and/or DPA. Of all sources of PUFAs the animal sources are preferably used, in particular fish and marine oil, or algae oil. In one embodiment of the present invention, the fish oil obtained from cod liver, herring, mackerel, salmon, menhaden and sardine is used as a source of LC-PUFA's.

According to the present invention and/or embodiments thereof, the fat is preferably present in a concentration from 0.01 to 80 wt.% in the food composition, more preferably from 0.1 to 60 wt.%, even more preferably from 0.2 to 50 wt%, even more preferably from 0.5 to 40 wt%, even more preferably from 0.8 to 35 wt%, even more preferably from 1.0 to 30 wt%, more preferably from 1.2 to 25 wt%, more preferably from 1.3 to 20 wt%, more preferably from 1.5 to 15 wt% and most preferably from 1.8-13 wt%. The fat is preferably present in the form of an emulsion in the composition of the invention.

Preferably, the emulsion is an oil-in-water emulsion. This is especially preferable in case butterfat is used as animal fat.

According to the present invention and/or embodiments thereof, the composition may further contain other vitamins and/or minerals, emulsifiers, colorants, preservatives, gums, thickeners. Where PUFAs are used in the composition, it is preferred to use antioxidants like vitamin C, vitamin E, selenium, polyphenols, bioflavonoids to prevent the oxidation of long chain fatty acids, which can lead to unwanted taste changes and a short shelf life of the product.

In a preferred embodiment of the present invention and/or embodiments thereof, the composition contains further vitamin D and, preferably, vitamin D3. The amount of vitamin D may range from 0.25 to 6 meg and preferably from 0.5 to 5 meg per daily intake, more preferably from 1 to 4 meg per daily intake, more preferably from 1.2 to 3.5 meg per daily intake, more preferably from 1.5 to 3 meg per daily intake, more preferably from 1.7 to 2.5 meg per daily intake, more preferably from 2 to 2.2 meg per daily intake. As used herein, "vitamin D" refers, to any of known form of vitamin D, and specifically includes vitamin D2 (ergocalciferol), vitamin D3 (cholecalciferol), vitamin D precursors, metabolites and another analogues, and combinations thereof, as well as the various active and inactive forms of vitamin D. For example, vitamin D3 may be provided in its unhydroxylated inactive form as cholecalciferol, or may be provided in its hydroxylated active form as calcitriol.

In a preferred embodiment of the present invention and/or embodiments thereof, the composition contains vitamin D and, preferably, vitamin D3 in the amount of from 0.25 meg to 15 meg per 100 gram or 100 ml, more preferably from 0.5 to 12 meg per 100 gram or 100 ml, more preferably from 1 to 10 meg per 100 gram or 100 ml, more preferably from 1.2 to 8 meg per 100 gram or 100 ml, more preferably from 1.5 to 6 meg per 100 gram or 100 ml, more preferably from 1.7 to 5 meg per 100 gram or 100 ml, more preferably from 2 to 4 meg per 100 gram or 100 ml, more preferably from 2.5 to 3.5 meg per 100 gram or 100 ml, and most preferably from 2.7 to 3.2 meg per 100 gram or 100 ml product.

In yet another preferred embodiment of the present invention and/or embodiments thereof, the composition contains vitamin B12 (various cobalamins). This vitamin contributes to energy metabolism, red blood cell formation, function of the immune system and mental health. Suboptimal blood levels are quite common in human and particularly in elderly, as absorption through the gut declines with age. Low vitamin B12 blood levels results in a macrocytic anemia, elevated homocysteine which increases the risk for cardiovascular diseases, peripheral neuropathy, memory loss and other cognitive deficits. A preferred amount for vitamin B12 is from 0.25 to 3 meg, and preferably from 0.375 to 2 meg per daily intake.

In a preferred embodiment of the invention and/or embodiments thereof, the composition may further comprise minerals such as calcium and/or magnesium and/or potassium. These minerals may have a beneficial effect in muscle function that results in the regulation of e.g. the heart beat, the diameter of arteries, blood pressure and/or blood flow. It is to be understood that any form of food grade sources of calcium, magnesium and potassium may be used. These include organic and inorganic compounds, salts, molecular complexes, chelates, such as calcium citrate, calcium carbonate, calcium gluconate, calcium lactate, calcium phosphate, magnesium oxide, magnesium citrate, etc. For magnesium, a preferred source is magnesium citrate. As to calcium, dairy products or dairy components, especially milk and milk parts, are examples of suitable sources of calcium. In a preferred embodiment of the invention and/or embodiments thereof, calcium (Ca) is preferably present in the composition of the invention in an amount from 20 to 1600 mg, and preferably from 70 to 1000 mg, more preferably, from 80 to 800 mg per daily intake. A preferred amount of Ca in the present composition is in the range of 100-600 mg per daily intake.

In a preferred embodiment of the invention and/or embodiments thereof, the amount of calcium is 20-4000 mg per 100 gram or 100 ml, more preferably 70-2500 mg per 100 gram or 100 ml, more preferably 80-2000 mg per 100 gram or 100 ml, more preferably, 100-1500 mg per 100 gram or 100 ml, more preferably from 150-1250 mg per 100 gram or 100 ml, more preferably from 200-1000 mg per 100 gram or 100 ml, more preferably 250-800 mg per 100 gram or 100 ml, more preferably from 300 to 600 mg per 100 gram or 100 ml, more preferably from 400-500 mg per 100 gram or 100 ml product.

The content of magnesium (Mg) is preferably from 20 to 800 mg, more preferably, from 30 to 500 mg, and even more preferably from 50 to 300 mg per daily intake. In a preferred embodiment of the present invention and/or embodiments thereof, the composition comprises 80-800 mg calcium and 50- 300 mg magnesium per daily intake.

In a preferred embodiment of the invention and/or embodiments thereof, the amount of magnesium is 20-2000 mg per 100 gram or 100 ml, more preferably from 30-1500 mg per 100 gram or 100 ml, more preferably from 50 to 1000 mg per 100 gram or 100 ml, more preferably from 80 to 750 mg per 100 gram or 100 ml, more preferably from 100 to 500 mg per 100 gram or 100 ml, more preferably from 150 to 250 mg per 100 gram or ml product. In a preferred embodiment of the present invention and/or embodiments thereof, the composition comprises 80-2000 mg calcium and 50-750 mg magnesium per 100 gram product or 100 ml product.

If the composition according to the invention is a dairy composition, in one preferred embodiment, the dairy component is the sole source of Ca, that is, the composition is not supplemented with additional Ca. Therefore, in this embodiment, the level of Ca corresponds to the level of Ca in milk, that is, from 50 to 200 mg per 100 g, or from 60 to 400 mg per serving. The Ca daily intake in this case is preferably from 80 to 800 mg, more preferably, from 100 to 700 mg, even more preferably from 120 to 600 mg, and particularly preferably from 240 to 500 mg. In another preferred embodiment of the present invention and/or embodiments thereof, however, the dairy composition can be supplemented with additional calcium so that the total amount of calcium present is, e.g., from 100 to 1200 mg, and preferably from 120 to 1000 mg and even more preferably from 240 to 800 mg per daily intake.

In another preferred embodiment of the present invention and/or embodiments thereof, however, the dairy composition can be supplemented with additional calcium so that the total amount of calcium present is, e.g., 100 to 3000 mg, more preferably from 120-2500 mg, more preferably from 240-2000 mg calcium per 100 gram or 100 ml product.

In another preferred embodiment of the present invention and/or embodiments thereof, the Ca/Mg ratio of the composition is not greater than 8 and more preferably not greater than 6, such as 5 or less. Yet more preferably, the Ca/Mg ratio is not greater than 3.5 and even more preferably, not greater than 3. A preferred range for the Ca/Mg ratio is 0.5-2.7. The ratio Ca/Mg in the range 1.5-2.4, for example, about 2, is particularly preferred. A low Ca/Mg ratio relates to each product or serving separately.

In another preferred embodiment of the present invention and/or embodiments thereof, potassium (K) is preferably present in the composition of the invention in an amount from 50 to 2000 mg per daily intake, more preferably from 100 to 1000 mg potassium per daily intake, more preferably from 150 to 800 mg K per daily intake, even more preferably from 200 to 700 mg K per daily intake and most preferably from 250-600 mg K per daily intake.

In another preferred embodiment of the present invention and/or embodiments thereof, the amount of potassium is 50-5000 mg, more preferably 1000-2500 mg, more preferably 150 to 2000 mg, even more preferably from 200-1750 mg, more preferably from 250-1500 mg, more preferably 300-1000 mg, more preferably from 400 to 800 mg, more preferably from 500 to 600 mg potassium per 100 gram or 100 ml product.

In another preferred embodiment of the present invention and/or embodiments thereof, the composition of the invention may further comprise at least one milk or milk-derived ingredient. In this case the composition is considered a dairy composition. These ingredients may be milk proteins such as casein and whey proteins, e.g. lactalbumin and lactoglobulin, including hydrolyzed casein and whey proteins, casemates, but also lactose, curd, milk powder, milk solids, butter solids, butter oil, butterfat, lactic acid, recaldent. Also milk and products based on milk or obtained by processing of milk such as dairy cream, cheese, milk powder, sauces, yogurt, fermented milk products are encompassed by the term dairy composition. In this embodiment, the fat may be butterfat and/or milk fat alone or in combination with another fat.

In a particularly preferred embodiment of the present invention and/or embodiments thereof, the food composition is a yogurt, which may be of spoonable consistency or in the form of a drink. The yogurt of the invention preferably contains 0.01-10 wt.% of fat, preferably from 1 to 5 wt.%, more preferably from 1 to 3 wt%, more preferably from 1.2 to 2 wt% and most preferably from 1.5 to 1.8 wt%, based on the total weight of the composition,

In another preferred embodiment of the present invention and/or embodiments thereof, the food composition is a cheese. Under cheese a milk fermentation product is meant that includes both soft and hard cheeses.

Examples of cheeses are Edam cheese, Gouda cheese, emmental, brie, camembert, cheddar, cottage cheese, low fat cheese, cream cheese, cheese spread, fresh cheese, mozzarella, feta, mascarpone, parmezan. A cheese of the invention preferably contains 3-75 wt.% of fat, preferably from 20 to 60 wt.%, based on the total weight of the composition. In yet another preferred embodiment of the present invention and/or embodiments thereof, the composition is a milk, cream or milk -based drink. Examples of these include whipping cream, fruit juice-containing milk drinks, fermented milk products, coffee milk, milk blends. The milk, cream or the milk -based drink of the invention preferably contains 0.05- 35 wt.% of fat, preferably from 0.1 to 30 wt.%, more preferably 0.5 to 25 wt%, more preferably 1 to 20 wt%, more preferably from 1.2 to 15 wt%, more preferably from 1.3 to 12 wt%, and most preferably from 1.5 to 10 wt% based on the total weight of the composition.

The composition may further be an ice-cream, acidified milk, whipped cream, buttermilk, cream -based sauces, condensed milk, pudding, sour cream, creme fraiche custard, and/or dairy spreads.

In another aspect the invention relates to a method of preparation of the composition according to the invention and/or embodiments thereof, comprising combining vitamin MK-7 and a fat comprising at least 36 wt% saturated fatty acids, and optionally adding the MK-7 to a food product. The composition according to the invention may be prepared in any convenient way, known to a person skilled in the art. In a preferred embodiment, the composition according to the invention is prepared by adding a suitable source of MK-7 and a suitable source of animal fat to a food product, preferably, to obtain a dairy composition. Under suitable source it is understood a source such as a compound or a composition comprising the desired vitamin or element, which source is suitable for use in food products (food grade).

Preferably, the suitable source contains MK-7 in a bioavailable form, such as in a chelated form or as an organic acid salt. Animal sources are preferred over plant sources. Suitably the MK-7 is added in a premix of vitamins and/or minerals. Also suitably the MK-7 is added to a fruit preparation and/or fruit syrup. The fruit preparation and/or syrup is then added to the food

composition. The ingredients can be mixed in any conventional way known to a person skilled in the art. For example, MK-7 can be dissolved in a fat phase comprising the fat, which fat phase can subsequently be emulsified with an optional aqueous phase and/or added to a food product to be fortified. MK-7 can also be used in powder form and be added to the remaining ingredients in an aqueous phase, or directly to the product.

In a further aspect the invention relates to a composition according to the invention and/or embodiments thereof to support and/or maintain and/or improve a condition selected from the group consisting of vascular health, cardiovascular health, macrocirculation of the blood, microcirculation of the blood, blood perfusion, vessel flexibility, bone health, clotting capability, platelet aggregation capabilities, glucose control, insulin secretion, insulin sensitivity, mental health, cognitive performance, memory, alertness, mental sharpness, weight management, triglyceride levels in blood, blood pressure, BMI, and/or waist circumference and/or to prevent diabetes, obesity, metabohc syndrome, arterial stiffness, chronic inflammation and/or low grade

inflammation.

In another aspect the invention relates to a composition according to the invention and/or embodiments thereof for use in the treatment of a condition selected from the group consisting of vascular disease, cardiovascular disease, atherosclerosis, clogged arteries, vessel inflexibility, varicose veins, transient ischemic attack (TIA), bone disease, bone demineralisation, osteoporosis, thrombosis, stroke, diabetes, insulin resistancy, high triglyceride blood levels, metabohc syndrome, obesity, bodily fat mass accumulation, high waist circumference, hypertension, chronic inflammation, low-grade

inflammation, mental disease, mental decline, dementia and/or Alzheimer disease.

Yet another aspect of the invention further relates to the use of a composition according to the invention and/or embodiments thereof to support and. or maintain and/or improve a condition selected from the group consisting of vascular health, cardiovascular health, vessel flexibility, blood pressure, blood flow, blood circulation, macrocirculation of blood, microcirculation of blood, blood perfusion, bone health, clotting capability, platelet aggregation capabilities, glucose control, insulin secretion, insulin sensitivity, mental health, cognitive performance, alertness, memory, and mental sharpness and/or to prevent a condition selected from the group consisting of diabetes, obesity, metabolic syndrome, arterial stiffness, chronic inflammation and/or low grade inflammation.

Another aspect of the invention relates to the use of a composition according to the invention and/or embodiments thereof to enhance

bioavailability of vitamin K and/or vitamin K2 and/or MK-7, and/or to increase the biological activity of vitamin K and/or vitamin K2 and/or MK-7, and/or the level of vitamin K and.or vitamin K2, and/or MK-7 in blood of a mammal.

It was surprisingly found by the present inventors that a composition comprising a fat that has at least 36 wt% saturated fatty acids the level of vitamin K in the blood is increased faster and to a higher level than when the vitamin K is in a composition with a fat having less than 36 wt% saturated fatty acids. The vitamin K supplements as used in the prior art mostly contain liquid vegetable oils such as sunflower, soy, natto, linseed, and/or olive oil. It was found that already at lower levels of vitamin K2, e.g. 56 meg per day, levels of vitamin K2 in the blood are reached that are normally reached when almost twice the amount of vitamin K2 is used. The invention allows to use lower doses of vitamin K2 to achieve the same effect as in the prior art. As vitamin K is involved in blood clotting, it is desirable to have lower doses of vitamin K.

Moreover another aspect of the invention relates to the use of a composition according to the invention and/or embodiments thereof to reduce the level of circulating uncarboxylated MGP and/or uncarboxylated osteocalcin in blood of a mammal. Another aspect of the invention relates to the use of a composition according to the invention and/or embodiments thereof to increase the level of circulating carboxylated MGP and/or carboxylated osteocalcin in blood of a mammal.

In a preferred embodiment of the present invention and/or embodiments thereof the reduction of the level of circulating uncarboxylated MGP and/or uncarboxylated osteocalcin, and/or the increase of the level of circulating carboxylated MGP and/or carboxylated osteocalcin in blood of a mammal and/or the increase of the level of vitamin K and/or vitamin K2 and/or MK-7 in blood of a mammal is obtained within 12 weeks, preferably within 8 weeks more preferably within 4 weeks.

The higher levels of vitamin K in the blood as obtained by the present invention lead to a decrease in uncarboxylated MGP and/or

uncarboxylated osteocalcin. The uncarboxylated form of MGP and osteocalcin are the inactive form of these proteins. The carboxylated form of MGP and osteocalcin are thought to be involved in cardiovascular health. Already after 4 weeks of administration of a composition of the present invention a significant decrease in uncarboxylated MGP and uncarboxylated osteocalcin was observed.

In a further aspect, the invention further relates to a method to reduce the level of circulating uncaboxylated MGP and/or uncarboxylated osteocalcin in blood of a mammal by administering to a mammal a composition according to the invention and/or embodiments thereof.

Furthermore, another aspect of the invention relates to a method to increase the level of circulating carboxylated MGP and/or carboxylated osteocalcin in blood of a mammal by administering to a mammal a composition according to the invention and/or embodiments thereof.

Yet an another aspect of the invention is directed to a method to increase the level of vitamin K, vitamin and/or K2 and/or MK-7 in blood of a mammal by administering to a mammal a composition according to the invention and/or embodiments thereof.

Preferably the composition according to the invention and/or embodiments thereof in a method and/or use according to the invention and/or embodiments thereof is administered for a period of at least 4 weeks, preferably at least 8 weeks and most preferably at least 12 weeks. Preferably the product of the invention is administered a person's whole life. Suitably, the product is administered for a period of 1, 2, 3, 5, 7, 10, 12, 15, 18, 20, 23, 25, 27, 30, 32, 35, 38, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or even up to 95 years.

The invention will now further be illustrated in the following, non- limiting examples.

Example 1: Long term intervention with yogurt drink and milk supplemented with vitamin K2.

In total 38 postmenopausal women received daily during one year two dairy products (1 portion of milk (250 ml) and 1 portion of yoghurt) with 1.5 wt% - 2 wt % milk fat (of which 66 wt % is saturated fat), circa 3 wt% protein and circa 10 grams carbohydrate and which were enriched with in total 100 meg vitamin K2 (MK-7), 10 meg Vit D3 and 800 mg Ca.

Blood samples were taken before and after the intervention period for vitamin K analysis. The circulating K vitamins were analysed in serum as described in Schurgers and Vermeer, Haemostasis, 2000: 30, 299-307. Briefly, serum samples were extracted with hexane and after a prepurification using silica columns they were analyzed by High Performance Liquid Chromatography (HPLC). For the HPLC, a reversed phase column with online zinc reduction and fluorescence detection was used and a synthetic form of K vitamin (vit Ki - 25) was used as internal control.

As a result of the intervention, the vitamin K2 level in blood is increased from 0.64 ng/ml (control level, before intervention), to 4.1 ng/ml. This vitamin K2 blood level after dairy consumption (4.1 ng/ml) is significant higher than expected based on published studies with intervention using vitamin K2 in soft gels (oil capsules that are low in saturated fat ; less than 16%). Namely, based on comparable vitamin K intake levels, these are in the range of 1.7 ng/ml - 3.3 ng/ml (Theuwissen, , et al, Br J. Nutr. 2012 page 1-5; Schurgers, et al, Blood 2007; 109: 3279-3283).

Example 2: Human intervention with yogurt drink supplemented with 56 meg vitamin K2 daily .

Twenty seven healthy men and postmenopausal women between 45 and 65 years of age have received daily 2 nutrient-enriched yoghurt products during 12 weeks (250 ml each, so 500 ml in total per day). Per 100 ml, the dairy matrix contained 3 gram protein, 1.5 gram of fat (of which 66 wt% is saturated fat), 10 gram of carbohydrates (lactose and added sugar). Moreover, per 100 ml the products were enriched with vitamin K2 in the form of MK-7 (11,25 microgram), omega-3 eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA); 40 mg EPA + DHA), calcium (120 mg), magnesium (56,25 mg), vitamin C (12 mg), and vitamin D3 (0.75 microgram).

Blood samples were taken at baseline and after 4, 8 and 12 weeks. Two weeks (14 days) before the start of the study until the final blood sampling,

participants were asked to restrict their intake of vitamin K-containing, namely no curd or cottage cheese, maximum of 1 piece of cheese per day (± 30 gram) and maximum of 200 gram of green vegetables. In this way, the normal daily consumption of vitamin K2 from other food products (estimated on 30 microgram) was kept constant. In addition, subjects were urged not to change their level of physical exercise or use of alcohol during the study. Further, the subjects were instructed to keep a stable body weight; therefore body weight was measured every study visit.

The circulating K vitamins were analysed in serum as described in Schurgers and Vermeer, Haemostasis; 2000: 30, 299-307. The results of this study were the following. The levels of vitamin K2 (MK7) in serum were measured to be 0.5-0.7 ng/ml before intervention and 2.7 ng vitamin K2 (MK-7) per ml serum after 12 weeks intervention. Based on the earlier study results with the known MK-7 formulation in oil capsules low in saturated fat (estimation of 1.4-1.7 ng/ml based on similar vitamin K2 amount in food), this is about 40%-50% higher than expected (Theuwissen,E, et al, Br J. Nutr. 2012; Schurgers, et al., Blood 2007; 109: 3279-3283). Moreover, the increased vitamin K2 (MK-7) serum levels after the intake of vitamin K2 enriched dairy products resulted in a significant efficacy of vitamin K2. This could also not been shown with the vitamin K2 oil capsules (<16% saturated fat), of the prior art. In the prior art, using low level of vitamin K2 oil capsules (45-90 microgram per daily dose), no significant effect after 3 months intake could be detected ((Theuwissen,E, et al., Br J. Nutr. 2012).

Efficacy was measured by means of the circulating biochemical markers desphospho-uncarboxylated MGP (dp-ucMGP) and uncarboxylated osteocalcin (ucOC) at various time-points (t = 0, 4, 8, and 12 weeks). Dp-ucMGP is a biomarker for vascular health and for arterial stiffness in particular, whereas ucOC has now been regarded as a more common health marker, including bone health, glucose control, lipid blood levels, weight management and metabolic syndrome. Vitamin K2 is required for activation of the protein ostecalcin. Active osteocalcin is implicated in bone mineralization and calcium homeostasis. It acts as a hormone and regulates e.g. the production of insulin, raises body's sensitivity to insulin and reduces stores of fat in a mammal.

Interestingly, the MK-7 enriched dairy product resulted in a significant and faster efficacy of vitamin K2 in comparison with the earlier documented vitamin K2 formulation in sunflower oil (see Table 1 and 2). The MK-7 enriched dairy product resulted in significant changes in ucOC and dp-uc MGP markers after 4 weeks with the low dosis of 56 meg MK-7: In contrast in the prior art for a MK-7 formulation in oil capsules (< 16% saturated fat), a higher dosage and/or longer exposure is needed to detect a significant effect

(Theuwissen,E, et al, Br J. Nutr. 2012).

Serum ucOC levels after intervention with dairy product enriched with vitamin K2 in healthy men and women (n=27)

*A significance of p<0.01 in comparison of serum levels at the start of the intervention period (t=0)

Serum dp-uc MGP levels after intervention with dairy product enriched with vitamin K2 in healthy men and women (n=27)

*A significance of p<0.01 in comparison of serum levels at the start of the intervention period (t=0) Example 3 : Intervention with yogurt drink or sunflower oil capsules

containing vitamin K2 in the form of MK-7.

A human intervention study with healthy men and postmenopausal women between 45 and 65 years of age which received ad random 50 meg of vitamin K2 (MK-7) in 3 types of matrix and 32-37 subjects per arm/in 105 subjects in total.

The different matrixes are

(1) MK-7 supplements (50 microgram) in sunflower oil capsules,

(2) MK-7 (50 microgram) in a yogurt drink (3 % protein, 1.5 % of fat (of which 66 wt% is saturated fat), 10 % of carbohydrates (lactose and added sugar) or

(3) MK-7 (50 microgram) in a yogurt matrix (3 % protein, 1.5 %of fat (of which 66 wt% is saturated fat), 10 % of carbohydrates (lactose and added sugars) with added omega-3 eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA); 40 mg EPA + DHA/100 ml), calcium (120 mg/100 ml), magnesium (56,25 mg/100 ml), vitamin C (12 mg/100 ml), and vitamin D3 (0.75 microgram/100 ml).

During 6 weeks, in total, daily 450 ml of the dairy drinks are consumed or MK- 7 capsules are consumed. Blood samples are taken at baseline and after 2 weeks, 4 weeks and 6 weeks. Also blood samples are taken after the

intervention at 3 days, t=l week and t=2 weeks (wash out period).

The circulating K vitamins, uncarboxylated osteocalcin (ucOC), carboxylated osteocalcin (cOC), (desphospho and uncarboxylated Matrix Gla Protein (dp-uc MGP) are analysed in serum as described in Schurgers and Vermeer,

Haemostasis; 2000: 30, 299-307.

As a result of the intervention, significant differences in serum MK-7 levels between dairy products and the supplements at week 4 have been detected. Moreover, the change of biochemical markers was dependent on treatment: for dp-uc MGP the change was 58 pM„ 73pM and 99 pM for respectively MK-7 supplements (group 1), MK-7 in yogurt (group 2) and MK-7 in yogurt matrix with extra nutrients (group 3). For cOC, MK-7 supplements did not show a treatment effect, whereas the two yogurt products were significant effective in raising vitamin K2 dependent carboxylation levels , Significant differences in cOC has been shown between group 3 (de dairy product with extra omega-3, vitamin D3, Ca Mg and vitamin C) and the MK-7 supplements. So, In line with previous studies, the dairy products enriched with MK-7, the dairy product with extra omega-3, vit D3, Ca, Mg and vitamin C in particular, give better results than the supplements of MK-7 dissolved in sunflower oil. Thus, MK-7 enriched product with high saturated fat levels gives better results in comparison with MK-7 supplements dissolved in sunflower oil (with a low (11%) level of saturated fat). These results show the effect of the food matrix, and particular the saturated fat content, on bioavailability and activity of vitamin K2, in particular MK-7.

Serum MK-7 levels changes in MK-7 levels (compared to baseline) and area under the curve (AUC) in ng/ml after intervention with supplement (group 1) and dairy products enriched with vitamin K2 in healthy men and women (group 2 and 3)

MK-7 levels (ng/ml) Group l:MK-7 Group 2: MK-7 Group 3: MK-7

supplements in yogurt drink yogurt matrix with

(n=37) (n=32) added omega-3

EPA/DHA, magnesium, vitamins C & D3 (n= 36)

Δ ΜΚ-7 t=2 2,09 ± 0,79 2,26 ± 0,89 2,35 ± 1,20

Δ ΜΚ-7 t=4 2, 19 ± 0,86*# 2,52 ± 0,93* 2,54 ± 1,27#

Δ ΜΚ-7 t=6 2,41 ± 1, 17 2,69 ± 1,20 2,73 ± 1,30

Δ AUC t=2 3,01 ± 1,04 2,97 ± 1,21 3, 12 ± 1,38

Δ AUC t=4 6,22 ± 2,24*# 6,46 ± 2,44* 6,61 ± 2,99#

Δ AUC t=6 9.99 ± 4,20 10, 19 ± 4,53 10,50± 4,60 *Between group differences (group 1 versus group 2) tested by linear regression with correction for baseline values, age , sex, BMI (p<0.05) #Between group differences (group 1 versus group 3) tested by linear regression with correction for baseline values, age , sex, BMI (p<0.05)

Levels of biochemical markers after different treatments of 6 weeks.

*Within group differences as compared to baseline tested by paired sample t test (p<0,05)

#Between group differences tested by ANOVA with Boniferroni correction (p<0.05) Figure 1 shows circulating dp-ucMGP levels (means with SE in pM) after intake of vitamin K2 suppletion via supplements (blue), via yogurt matrix with extra nutrients including omega-3, vitamin C, vitamin D3, calcium and magnesium (red), and via yogurt drink (green).

Figure 2 shows the level of circulating cOC (means with SE in ng/ml) after intake of vitamin K2 suppletion via supplements (blue), via yogurt matrix with extra nutrients including omega-3, vitamin C, vitamin D3, calcium and magnesium (red), and via yogurt drink (green).