WO2017098089A1

WO2017098089A1 - The use of a composition for lowering cholesterol level in a mammal, a method for its preparation, a composition and a method for preparing food additive comprising said composition

Info

Publication number: WO2017098089A1
Application number: PCT/FI2016/050864
Authority: WO
Inventors: Thomas Ahlnäs
Original assignee: Granula Ab Ltd
Priority date: 2015-12-09
Filing date: 2016-12-09
Publication date: 2017-06-15
Also published as: MX2018007065A; CA3008075A1; EP3386524A1; CN108601803A

Abstract

The invention relates to the use and method for preparation of a composition comprising at a range of 1–50 wt-% preferably at a range of 5–50 wt-% a lignan mixture comprising fatty acids, resin acids, lignans and oligomeric lignans, wherein lignans comprise at least following lignans: 7- hydroxymatairesinol, conidendrin, lariciresinol, liovil, secoisolariciresinol or their geometric isomers or stereoisomers thereof and wherein the amount of oligomeric lignans is 10 - 50 % of the total weight of said lignan mixture, for lowering the cholesterol level of blood serum of a mammal.

Description

The use of a composition for lowering cholesterol level in a mammal, a method for its preparation, a composition and a method for preparing food additive comprising said composition

The present invention relates to use according to claim 1 of a composition comprising a lignan mixture for lowering cholesterol level of blood in a mammal.

The invention relates also to a method for preparing a composition for lowering cholesterol level of blood serum in a mammal according to claim 12.

Additionally, the invention relates to a composition for lowering cholesterol lev- el in a mammal comprising said lignan mixture.

The invention further relates to a method according to claim 25 for preparing food additive, a dietary supplement or a nutraceutical for lowering cholesterol level of blood serum in a mammal.

In the field of food products there is a continuous demand for new dietary sup- plements having beneficial effect on human health. This demand arises on one hand from the fact that food products are often so called convenience food products which lack many essential nutrients and may contain additionally modified fatty acids, so called trans-fatty-acids, which have unfavourable effect on triglyseride levels and content of blood serum. On the other hand a big chunk of modern diet contains animal based food products (meat, milk, butter, cheese etc.) which contain saturated fats having unfavourable effect on blood triglyseride content and which increase blood cholesterol level.

Plant lignans are the primary source of dietary phytoestrogens in modern diet. Phytoestrogens have multitude beneficial health effects; high levels of lignans can support for example healthy glucose metabolism and reduce insulin sensitivity. They have also suggested to be used for treating general infections and inflammations (see US 5762935) when used as dietary supplement.

However, plants contain so low levels of lignans that their health effects in normal diet are almost insignificant. The lignan content of foods is generally low and usually does not exceed 2 mg/100 g.

Wood lignans are very abundant in wood material and bark and thus one might consider using these lignans in the diet in the same way as plant lignans. It has been suggested to use wood lignans in food based on their antioxidative effect and some wood lignans science have also been found to be effective against cancer(s). However, wood lignans and extracts containing wood lignans have usually a dark colour and a bitter aftertaste, which have effectively prevented their usage as a food additive.

For these reasons wood lignans have been used only in the field of cosmetics, which use is based mainly on their antioxidative and antimicrobial effect. For example, the applicant has earlier presented some wood lignan mixture(s) to be used in various cosmetics products to enhance the effect and safety of UV- protective agent used therein.

Based on the above mentioned prior art the inventors set a task to study the use of wood lignans especially in the diet for gaining new healthy food products.

Therefore, the main objective of the present invention is to find new uses for wood lignans.

The second objective of the invention is to find new uses for wood lignans in spite of their somewhat negative properties as to their colour and taste.

The third objective of the invention is to find a simple method for preparing lignan based ingredients to be used as food additives and food grade components, which can be added into food products.

These and other objects and features of the invention will be apparent from the following description and the claims.

Firstly, the present invention is based on the very surprising observation that certain lignan mixture comprising at least following lignans: 7-hydroxy- matairesinol, conidendrin, lariciresinol, liovil and secoisolariciresinol or their geometric isomers or stereoisomers thereof, have a very significant blood cho- lesterol lowering effect when these lignan mixtures are used as a dietary supplement for a mammal. The concentration of lignan mixture should be 1-50 wt-% of said compound used as a food additive. Preferably the concentration of lignan mixture is 5-50 wt-% of the total weight of a food additive.

Due to its considerable blood cholesterol lowering effect, the wood lignan mix- ture can be used in relatively small quantities; normal daily dietary portion of lignan mixture is 30-50 mg. When the lignan mixture is used as an alcohol- wood extract or alcohol-water-wood extract containing 1-5 % lignan mixture, its daily usage can be kept as low as 5-10 droplets.

The present invention is very surprising because this combination of wood lignans has not known to have any blood cholesterol effect and since this effect is unmatched compared to known plant lignans; daily diet of most humans include plant lignans which have similar properties, for example antioxidative properties as wood lignans. However, the level of lignans is usually very low in edible plants and therefore their possible beneficial cholesterol lowering effect has not been emergenced and the suggested usage level of plant lignans in food additives is also comparatively high.

Secondly, the present invention is based on the fact that the phenolic compounds, such as lignans, tannins and flavonoids contained in the wood material, are extracted from the wood with an extraction medium, which as such is suitable for various food products. Extraction medium may be an extraction solution or subcritical or supercritical extraction fluid, such as CO2. In the previous practice of the art, lignans, tannins and flavonoids have been purified before using them in food industry.

Differing from said previous practice in the art, the invention is based on the fact that the raw extraction solution or extraction fluid is not purified before use. The raw extraction solution - wood extract or raw extraction fluid-wood extract is recovered from the wood material, such as wood chips or chipped wood pulp of the chemical pulp industry, which contain lignans and/or flavonoids and/or tannins as active ingredients and is then used as such for preparing various food products, food additives or semi-finished products usable in food products.

The food products, food additives or semi-finished products usable in food products can be recognized of being produced by the method according to invention or being the food product, food additive or semi-finished product of the present invention if they have an elevated level of lignan mixture originated to a wood material, which lignan mixture include oligolignans 10 -50 % . In case this composition according to invention is a solution composing of extraction liquid-wood extract, the relative amounts of active ingredients in lignan mixture may be modified for better suiting in a designed use. This can be made, for example, by a fractionating extracting procedure. The solvent to be used as an extraction solution may simply be selected on the basis of its suitability to further use.

This provides the advantage that the isolation of the raw extraction solution - wood extract or raw extraction fluid-wood extract from the wood material is simple and requires a considerably smaller number of process stages than be- fore.

The raw extraction solutional alcohol -wood-extract may be a mixture that originates in two or more tree species, such as spruce and pine or conifer and birch. The wood may originate from trees belonging to genus Picea or genus Abies. The wood may originate specifically from Picea sp., Tsuga sp. (hem- lock) or Acacia sp. Additionally Betula sp. may be used. The types and amounts of lignans obtained from different wood species vary considerably.

By adding lignans from other wood species or plants one might modify the effect of lignan mixture (=active ingredient).

Thirdly, the invention is based on the fact that the selection of active ingredi- ents incorporated into food additives or food grade components should be designed so, that they can affect to various human body parts. Human body contains various polar, semi-polar, and non-polar parts, organs and membranes. The passing of a specified active ingredient into human parts, organs by diffusion depends on its concentration. If there is only one kind of active ingredient present, it has one concentration which will define its passing. Therefore the active ingredient should contain a mixture of various lignans, each having different polarity and concentration. Examples of the various objects of the body, which require active ingredients that have different migration (concentration): cell wall (semi-polar), the exterior of the cell, water phase (polar), and hydro- phobic (the body parts that contain fat).

The present lignans seem to lower cholesterol level by preventing inflammation of blood vessel. If only one purified lignan compound would be used as the ac- tive ingredient, its migration into walls of blood vessels in the different places of the body would be considerably weaker than those of the active ingredient mixture (=lignan mixture) according to the invention.

Therefore, the invention according to claim 1 relates to the use of a lignan mix- ture for lowering cholesterol level of a blood serum in a mammal such as human.

To be more exact, the present invention relates to the use of a composition comprising at a range of 1-50 wt-% preferably at a range of 5-50 wt-% a lignan mixture comprising fatty acids, resin acids, lignans and oligomeric lignans, wherein lignans comprise at least following lignans: 7- hydroxymatairesinol, conidendrin, lariciresinol, liovil, secoisolariciresinol or their geometric isomers or stereoisomers thereof and wherein the amount of oligomeric lignans is 10 - 50 % of the total weight of said lignan mixture, for lowering the cholesterol level of blood serum of a mammal. One important aspect of the invention is also to make wood lignans available in such a level in an edible food product that their concentration is high enough to ensure that their beneficial blood serum cholesterol level lowering effect will appear. Normal wood material contains only small amounts of wood lignans and making food supplements from this kind of wood material would be cum- bersome and ineffective. However, wood bark and knot wood have been founded to include much higher amounts of these lignans which make mentioned lignan mixture.

Therefore, one important aspect of the invention relates to a method for preparing composition for lowering cholesterol level in a mammal blood serum us- ing wood bark or knot wood as source of lignans.

The invention relates also to a method according to claim 12 for producing a composition comprising said lignan mixture for lowering cholesterol level of a blood serum in a mammal.

To be more exact, the present method for producing a composition for lower- ing cholesterol level of blood serum in a mammal comprises: - extracting knot wood and/or bark with an extraction solution selected from the group composed of alcohol and alcohol-water mixture,

- obtaining an alcohol-wood extract or an alcohol-water-wood extract, and

- adding a possible edible carrier agent to said alcohol-wood extract or alcohol- water-wood extract. Said composition to be produced comprises at a range of

1-100 wt-% preferably at a range of 5-90 wt-% of lignan mixture and an extraction solution wherein said lignan mixture in extraction solution comprises fatty acids, resin acids, liovil, lignans and oligomeric lignans, wherein lignans comprise at least following lignans: 7-hydroxymatairesinol, conidendrin, lariciresinol, secoisolariciresinol or their geometric isomers or stereoisomers thereof and wherein the amount of oligomeric lignans or their geometric isomers or stereoisomers thereof is 10 - 50 % of the total weight of said lignan mixture present in said composition, wherein the extraction solution is selected from the group consisting of alcohol and water-alcohol mixture, the balance being a physiologically acceptable carrier agent.

The percent ranges given for lignan mixture means percent by weight, based on the total composition.

Lignans means herein normal dimeric lignans as defined in lUPAC 2000. Additionally liovils are herein included into lignans. Dimeric lignans have three or four phenylpropane units.

Oligomeric lignans, which are also referred as oligolignans, are trimeric or higher lignans. The definition of oligomeric lignans is herein same as presented in lUPAC 2000. This means, that oligomeric lignans may be sesquilignans, dilignans, sester-, tri-, sesquar- tetra- and pentalignans. For example trimeric lignans, so called sesquilignans, have an additional guaiacylglycerol unit bound to one of the phenolic units of lignans.

Resin acids are those mostly originated to pinus wood and contained palustric acid, abietic acid, dihyrdoabietic acid, pimaric acid, sandsrocopimaric acid, isopimaric acid, neoabietic acid and abietal. The most common resin acids are abietic acid and dehydroabietic acid, which are typically found from wood of Pinus species. The percent ranges (wt-%) given for lignan mixture mean percent by weight, based on the total product composition.

The present invention relates also to a food additive, nutraceutical or dietary supplement for lowering cholesterol level of blood serum in a mammal. To be more exact, the present invention relates to a method for preparing food additive, a dietary supplement or a nutraceutical for lowering cholesterol level of blood serum in a mammal, wherein said method comprises the following steps:

- extracting knot wood and/or bark of wood with an extraction solution select- ed from the group composed of alcohol and alcohol-water mixture for obtaining an alcohol-wood extract or an alcohol-water-wood extract,

- adding a possible edible solid or liquid carrier agent such as oil or aquous liquid or emulsion to said alcohol-wood extract or alcohol -water-wood extract and -mixing said alcohol-wood extract or alcohol-water-wood extract, in combination with a possible edible carrier agent to a food additive, dietary supplement or nutraceutical to prepare an edible mixture which is a food additive, a dietary supplement or a nutraceutical comprising at a range of 1-50 wt-% preferably at a range of 5-50 wt-% a lignan mixture comprising fatty acids, resin acids, lignans and oligomeric lignans, wherein lignans comprise at least following lignans: 7-hydroxymatairesinol, conidendrin, lariciresinol, liovil, secoisolar- iciresinol or their geometric isomers or stereoisomers thereof and wherein the amount of oligomeric lignans is 10 - 50 % of the total weight of said lignan mixture. Preferably said food additive, dietary supplement or nutraceutical comprises at a range of 1-90 wt-% preferably at a range of 5-90 wt-% said lignan mixture in a combination with an extraction solution selected from the group composing of alcohol and water-alcohol mixture the balance being an edible solid or liquid carrier material. The spectrum of effect of lignan mixture can be widened by adding other wood lignans or triterpens having for example different antimicrobial potency than said lignan mixture.

Therefore, the invention also relates to the use of composition further compris- ing stilbenes or triterpens selected from the group composed of betulin, betu- lonic acid, betulinic acid, betuloinic acid, resveratrol and their geometric isomers or stereoisomers thereof. The invention relates also to corresponding food additives, nutraceuticals or dietary supplements and products containing this kind of stilbenes or triterpenes. In this connection it should also be noted, that the lignan mixture itself has antimicrobial potency. Therefore, the invention also relates to extend the shelf life of a food material by mixing said food material with a food additive containing above mentioned lignan mixture obtained from wood material.

Furthermore, the composition according to present invention may also contain other lignans obtained from other sources such as various plant lignans. Additionally, composition can contain antioxidants such as vitamins, or substances for treating illnesses or to be used as microbicides (betulinic acid or betuline obtained from birch). When this kind of compounds are used in food products they can obtain new properties; they are either better for human health or they might even be helpful in preventing illnesses (so called health food). By extracting of different types of wood or plants, active ingredient mixtures are obtained having different properties, such as antioxidative, microbicidal and, possibly, disease-preventive, and abilities to catch free radicals.

It is known that lignan glycosides are absorbed in the gastrointestinal tract af- ter metabolism by intestinal bacteria to lignan aglycones and the enterolignans (enterolactone and enterodiol), which are formed from them. Both enterolac- tone and enterodiol have been shown to possess weakly estrogenic and antiestrogenic activities, and it has been suggested that the high production of these antiestrogenic mammalian lignans in the gut may serve to protect against breast cancer in women and prostate cancer in men. Thus the composition of this invention may further be used as a prophylactic composition to prevent development of cancerous diseases. A food additive preferably formulated so that lignan mixture and a carrier material form together an encapsulated liquid, powder or pill.

Typically the food additive according to present invention is added into various oils such as sauces, salad dressings or marinades. These food additives can also be mixed into various convenience foods and ready meals which may additionally be based on meat. Often these convenience foods or ready meals comprise so called "junk food" such as hamburgers, French fries etc. which have low level of nutrients and lot of modified fatty acids, so called trans-fatty-acids and/or saturated meat based fats. By adding food additives or dietary supplements comprising mentioned (wood) lignan mixture these "junk foods" are a bit healthier.

The food additive according to invention may be added also into plant based food products which have none or very few plant lignans. These are for example rice, pasta, macaroni and bread. If the food additive according to present invention is added into bread one can raise its health value but only its shelf life. The latter effect is of big significance because adding shelf life of bread will positively influence the whole distribution logic.

The food additive according to present invention can also be added into vari- ous drinks, juices and nectars.

The food additive according to present invention may also have beneficial effect if one wishes a bit bitter taste to food.

Preferably the lignan mixture used in the products, compounds and food additives according to the invention is obtained by extracting or grinding knot wood of coniferous trees, which belong to wood genus picea, abies, fir or spruce. For example in Norway spruce (Picea Abies) 7-hydroxymatairesinol is the dominant lignan but in other fir /Abies trees contain secoisolaricinol or lariciresinol as the main lignans (see also Knots in three -A new source of lignans, Holmbot et al.) phytochemistry Reviews 2: 331 -340, 2003). If this lignan mixture has been obtained from knot wood of genus picea, abies, fir or spruce, its lignan mixture has the following formula:

- 7-hydroxymatairesinol 70-80 %;

- conidendrin 3-8 %; - lariciresinol 1-4 %;

- liovil 2-5 %;

- secoisolarisinol 3-7 %

- other lignans 0-3 %.

^■oligolignans 10 -50 %, preferably 20 -50 %. Depending on the wood source of the alcohol-wood-extract, there may be also significant amount of other dimeric lignans present in each alcohol-wood- extract. For example knots of pine tree are known to contain plenty of nortrachelogenin, pinosylvin monomethyl esters and pinoresinol.

Depending on the source of wood material it may contain also other wood lignans such as pinoresinol, nortrachelogenin, matairesinol and juvabiones.

Although dietary plants may contain some lignans the level of lignans are usually very low.

On the other hand, if a food composition is prepared by adding composition containing purified lignans these compositions will not contain oligolignans or their level is extremely low.

For extracting of knot wood and/or bark with an extraction solution, for obtaining an alcohol-wood extract or an alcohol-water-wood extract, various alcohol- based solvents, such as butylene glycol, butylene glycol + monoalcanol, glycerol or glycerol + alcohol can be used. These alcohol based solvents are either physiologically acceptable or/and edible.

The extraction solution preferably contains a lower monovalent or bivalent alkyl alcohol or a mixture of the lower monovalent and bivalent alkyl alcohols and/or glycerol, or a mixture of the lower (monovalent) alkyl alcohol and the bivalent or a trivalent lower alkyl alcohol.

The monovalent alcohol is preferably ethanol, propanol, butanol, heptanol, oc- tanol or decanol. The monoalcohol is especially preferably ethanol. The bivalent alkyl alcohol is preferably a lower alkylene glycol, which is preferably selected from a group composing of ethylene glycol, propylene glycol, bu- tylene glycol, pentylene glycol, and dipropylene glycol. Butylene glycol is an especially preferable solvent, as the lignan mixture according to the invention dissolves therein in amounts of over 10% and it is physiologically tolerable. So- lutions of ethanol and bivalent alcohols are preferable for dissolving and also extracting the lignan mixtures according to the invention. Bivalent alcohol is especially preferably selected from the group composing of ethylene glycol, propylene glycol, butylene glycol, pentylene glycol, dipropylene glycol.

Suitable trivalent alcohol solvents are glycerols. The mixture of the lower alkyl alcohol and glycerol or alkyl glycol is a preferable solvent medium, as the need to further process the raw extracts (= solvent + active ingredient mixture) or raw extract concentrates based on extraction solutions containing such alcohols is as small as possible, when preparing different food additives or food grade compositions. The alcohol or water-alcohol based raw alcohol-wood extracts or alcohol-wood extracts are well-suited to the manufacture of food-grade components or to be used as such. These food grade components include, without limiting to them, emulsions, dispersions, oils, sweeteners.

Lignan mixtures of the present invention can be added as an alcohol-wood ex- tracts or alcohol-water-wood extracts into various food additives or food grade compositions made of these extracts.

Food additives are selected from the group composing of feed ingredients, food supplements and food additives for direct human consumption

A food supplement includes vitamin and mineral supplements in unit dose forms such as capsules, tablets, powders, solutions etc. This page provides in- formation on the food additive provisions that are acceptable for use in foods conforming to the food category. A comprehensive list of possible food supplements in which the iignan mixture of the present invention can be used, has been presented in GSFA online Food Category. A feed ingredient is a component part or constituent or any combination/mixture added to and comprising the feed. Feed ingredients might include grains, milling byproducts, added vitamins, minerals, fats/oils, and other nutritional and energy sources. A comprehensive list of feed ingredients has been presented in the Official Publication of the Association of American Feed Control Officials (AAFCO).

A comprehensive list of food additives in which the present Iignan mixture can be included has been presented in CFR Code of Regulations Title 21. These include food preservatives (subpart A), food coatings (subpart C), special dietary and nutritional additives (subpart D), anticaking agents (subpart E), fla- vouring agents and related substances (subpart F), gums, chewing gum bases and related substances (subpart G), other specific usage additives (subpart H), multipurpose additives (subpart I).

Some examples of food grade compositions or food additives into which the alcohol or water-alcohol based raw alcohol-wood extracts or alcohol-wood ex- tracts can be mixed and which belong to above mentioned multipurpose additives (subpart I) of CFR Code of Federal Regulations Title 21 :

172.809 Currdlan.

172.810 Dioctyl sodium sulfosuccinate.

172.81 1 Glyceryl tristearate.

172.812 Glycine.

172.814 Hydroxylated lecithin.

172.816 Methyl glucosidecoconut oil ester.

172.818 Oxystearin.

172.820 Polyethylene glycol (mean molecular weight 2009, 500).

172.822 Sodium lauryl sulfate.

172.824 Sodium mono and dimethyl naphthalene sulfonates.

172.826 Sodium stearyl fumarate.

172.828 Acetylated monoglycerides. 172.829 Neotame.

172.830 Succinylated monoglycerides.

172.831 Sucralose.

172.832 Monoglyceride citrate.

172.833 Sucrose acetate isobutyrate (SAIB).

172.834 Ethoxylated mono and diglycerides.

172.836 Polysorbate 60.

172.838 Polysorbate 65.

172.840 Polysorbate 80.

172.841 Polydextrose.

172.842 Sorbitan monostearate.

172.844 Calcium stearoyl-2-lactylate.

172.846 Sodium stearoyl lactylate.

172.848 Lactylic esters of fatty acids.

172.850 Lactylated fatty acid esters of glycerol and propylene glycol.

172.852 Glyceryl lacto esters of fatty acids.

172.854 Polyglycerol esters of fatty acids.

172.856 Propylene glycol mono and diesters of fats and fatty acids.

172.858 Propylene glycol alginate.

172.859 Sucrose fatty acid esters.

172.860 Fatty acids.

172.861 Cocoa butter substitute from coconut oil, palm kernel oil, or both oils.

172.862 Oleic acid derived from tall oil fatty acids.

172.863 Salts of fatty acids.

172.864 Synthetic fatty alcohols.

172.866 Synthetic glycerin produced by the hydrogenolysis of carbohydrates

The present lignan mixture of the present invention can also be mixed with carrier or extraction solvents which are appropriate food additives. The lignan mix- ture can be either powder or alcohol- water-wood extract or alcohol-wood extract.

A comprehensive list of these carrier or extraction solvents can be found from health Canada, Food and Nutrition. These carrier or extraction solvents include following: acetone, benzyl alcohol, 1 ,3-butylene glycol, carbon dioxide, castor oil, citric acid esters of mono-and diglycerides, ethyl acetate, ethanol, glycerol (glycerin), glyceryl diacetate, glyceryl triacetate, glyceryl tributyrate, hexane, isopropanol alcohol, methyl alcohol, methyl ethyl ketone(2-butanone), methylene chloride, mono- and diglycerides, monoglyceride citrate, 2-nitropropane, 1 ,2-propylene glycol (1 ,2-propanediol), propylene glycol monoesters and diesters of fatforming fatty acids, triethyl citrate,

Examples Example 1 A concentrated alcohol- wood extract containing following lignan mixture was prepared by extracting chipped knot wood of Picea Abies with an extraction solution composing of propylene glycol. The lignan extract contained 7.5 wt-% lignan mixture presented in table 1 :

Several other concentrated alcohol-wood-extracts were also prepared by ex- tracting chipped knot wood of Picea Abies with an extraction solution composing of butylene glycol, pentylene glycol or glycerine. The most important lignans present in these alcohol-wood-extracts extracted from Picea abies knots are same as presented in table 1 (Liovil is not actually a lignan but it is included herein in the table 1 ) Table 1

Lignan Amount, % W

Hydroxymatairesinol 5-6

Secoisolaricesinol 0,2-0,4

Conidendrin 0,2-0,4

Liovil 0,2-0,4

Lariciresinol 0,1-0,2 Other lignans 0,2-1

As can be seen from table 1 , there are a wide variation in lignan composition from extract to extract and the amount of "other lignans" in alcohol-wood- extracts may be quite significant. In the alcohol-wood-extract of figure 1 the main source of wood material (wood knots) was Picea Abies but there were admixed also a significant amount of wood material (wood knots) from pine tree, which brought also other lignans except those shown in table 1 in this alcohol-wood-extract.

To get a better conception of the composition of the lignan mixture these wood-alcohol-extracts were further analysed by using a several chromatographic methods. The composition of lignan mixture present in these extracts was first analysed by using gas chromatographic methods and results have been shown in figures 1 -3. The inner reference was betulinol.

It was discovered that actually each wood-alcohol-extract may contain except about 56 % normal dimeric lignans presented in table 1 (hydroxymatairesinol, secisiolarisinol, conidendrin, lariciresinol) but also about 10 -50 % usually 20 - 50 % oligolignans and 1 -4 % resin acids and fatty acids. As said before, although Liovil is not itself considered as a lignan (see lUPAC 2000 for lignans) it is here considered to be a part of the lignan mixture, for the simplicity of refer- ring.

Figure 1 shows results of a HPLC analysis for wood-alcohol-extract (Norway spruce with fractions of pine wood). This extract was added subsequently into various products in examples. The wood chips from knotwood of Norway spruce was first extracted with alcohol (ethanol) and from wood-alcohol-extract was identified the most important compounds with a HPLC-analysis. The inner standard in this HPLC-analysis was Betulin.

As can be seen from picture 1 , the analysed wood-alcohol-extract contains fatty acid resin acids, normal lignans and oligomeric lignans (oligolignans). The oligolignans (oligomeric lignans) were not qualified very deeply but this fraction seems to contain at least sesquilignans and dilignans but also sester-, tri, - sesquar- and tetralignans, tetralignan esters and pentalignans (definition and structure for these oligolignans is same as presented in lUPAC 2000). Most abundant was dilignans, sesquilignans and sesterlignans.

The fatty and resin acids and normal lignans were further analysed by HPLC, results are shown in figs. 2 and 3. Below is presented a summary of results and discussion.

Resin acids were mostly originated to pinus wood and contained palustric acid, abietic acid, dihyrdoabietic acid, pimaric acid, sandsrocopimaric acid, isopi- maric acid, neoabietic acid and abietal. However, the most abundant resin ac- ids were abietic acid and dehydroabietic acid which are typical resins found from wood of Pinus species.

Resin acids contained some resin acids originating to pine wood such as pi- nosylvin methyl ester.

The lignans mainly composed of 7-hydroxymatairesinol but significant amount of matairesinol, iso-Hydroxymatairesinol, isolaricinol, lariciresinol, conidendrin, conidendrin acid, secoisolariciresinol and liovil (todolactol) was present. The li- ovil is actually not a lignan. Because wood material contained traces from Pine wood, pinoresinol was also present.

Example 2

A concentrated alcohol-wood-extract containing a lignan mixture presented in table 1 was prepared by extracting chipped knot wood of Picea Abies with an extraction solution composing of ethanol and water. The alcohol-water wood extract contained 75 wt-% of lignan mixture presented in table 1 . This concentrated alcohol-wood-extract can be used as such, in preparing various alcoholic drinks. Each drink could include for example 10 g of above alcohol-water wood extract contained 75 wt-% of lignan mixture. Preferable alcoholic drinks are those having a little bitter aftertaste such as gin. Example 3

A food additive was prepared containing 80 wt-% of caprylic/capric monodi- glyceric acid (CAS 85536-07-8) and 20 wt-% of alcohol-water wood extract prepared in example 2. Thus food additive contained 15 wt-% of lignan mix- ture from its dry weight. The lignan mixture present in food additives has the same lignan composition as the lignan mixture mentioned in table 1 . This lignan mixture has about 25 -46 % of oligomeric lignans and the rest were ol- igmeric lignans, fatty acids and resin acids shown in figure 1 .

Example 4 An alcoholic composition (liquid) was prepared by mixing 10 weight-% of lignan extract prepared according to example 1 with 90 weight-% of 1 ,3-propanediol (Zemea, Dupont-Tate-Leyle). This composition was administered daily to patients 5 -10 drops/day.

Table 2 results:

It is evident from the results that the cholesterol, LDL, HDL and triglyseride levels decreased markedly after using five drops of the composition daily for a period of 3-12 months. Example 5

In a corresponding way as in example 3 can be prepared other food additives mentioned above. Lecithin can be used as a dispersion agent.

Examples 6-xx The following exemplary food additives, dietary supplements or nutraceutical foods were prepared while providing them with a lignan mixture.

Example 6a

Preparing nutraceutical dairy products

Wood lignan enhanced dairy products such as wood lignan enhanced milk, cheese, cream, creme fraiche, ice-cream, kefir, smetana, sour milk, yoghurt and viili may be prepared by simply adding wood lignan containing solutions straight into finished or semi-finished dairy products.

The preparation of enhanced dairy product may be performed as follows:

- extracting knot wood and/or bark with an extraction solution selected from the group composed of alcohol and alcohol-water mixture for obtaining an alcohol- wood extract or an alcohol-water-wood extract;

-admixing a physiologically acceptable carrier agent to said alcohol-wood extract or alcohol-water-wood to obtain a mixture containing alcohol-wood extract with a carrier agent or an alcohol-water-wood extract with a carrier agent; -admixing obtained mixture of alcohol-wood extract with a carrier agent or mixture of alcohol-water-wood extract with a carrier agent with dairy products.

The mixture of alcohol-wood extract with a carrier agent or mixture of alcohol- water-wood extract with a carrier agent may include lignan mixture 50-90 wt % calculated from said mixture. This mixture is added about 5-15 g per 100 g of dairy product.

Yoghurt, prepared from plain milk has typically a following composition:

Proximates per 100 g of yoghurt

Water 87 g

Energy 61 kcal

Protein 3.47 g

Total lipid (fat) 3.25 g Carbohydrate, 466 g

Fiber, total dietary 0 g

Sugars, total 4,66 g Minerals

Calcium, Ca 21 mg

Iron, Fe 0.05 mg

Magnesium, Mg 12 mg

Phosphorus, P 95 mg

Potassium, K 155 mg

Sodium, Na 46 mg

Zinc, Zn 0.59 mg

Vitamins

Vitamin C, total ascorbic acid 0.5 mg

Thiamin 0.029 mg

Riboflavin 0.142 mg

Niacin 0.075 mg

Vitamin B-6 0.032 mg

Folate, DFE 7 ug

Vitamin B-12 0.37 mg

Vitamin A, RAE 27 ug

Vitamin A, IU 99 IU

Vitamin E (alpha-tocopherol) 0.06 mg

Vitamin D (D2 + D3) 0.1 ug

Vitamin D 2 IU

Vitamin K (phylloquinone) 0.2 ug

Lipids

Fatty acids, total saturated 2.0 g

Fatty acids, total monounsaturated 0.89 g

Fatty acids, total polyunsaturated 0.09 g

Cholesterol 13 mg

Wood lignans (table 1 ) 5 g

Also other liquid or semi-liquid dairy products have similar or nearly similar composition as to minerals, vitamins, lipids and proximates. Dairy products do not contain any wood originated or even plant originated lignans. Therefore, all wood lignans present in any dairy product are brought by admixing wood lignans to said dairy product. Example 6b

Roll, hamburger or hotdog with wood lignans Hamburgers, rolls and similar products can be provided with wood lignans hav^¬ ing cholesterol lowering effect.

A typical nutrient value of a hamburger per 10Og:

Proximates

Water 34.34 g

Energy 279

Protein 9.77 g

Total lipid (fat) 3.91

Carbohydrate 50.12 g

Fiber 1 -8 g

Sugars, total 7.28 g

Vitamins

Vitamin C 1 .3 mg

Thiamin 0.543 mg

Riboflavin 0.297 mg

Niacin 4.180 mg

Vitamin B-6 0.063 mg

Folate, DFE 131 ug

Vitamin B- 2 0.20 ug

Vitamin A, RAE 29 ug

Vitamin A, IU 107 IU

Vitamin E 0.27 mg

Vitamin D 0 IU

Vitamin K 4.8 IU As can be seen above, a typical composition of hamburgers, rolls and similar products is normally loaded with carbohydrates and fat. By adding wood lignans 15 -30 g per product these products can be made a bit healthier or at least cholesterol level of a human can be controlled although these products are consumed.

These products can be prepared for example by:

-extracting knot wood and/or bark with an extraction solution selected from the group composed of alcohol and alcohol-water mixture,

-obtaining an alcohol-wood extract or an alcohol-water-wood extract, which have a composition comprising at a range of 1-90 wt-% preferably at a range of 5-50 wt-% a lignan mixture comprising fatty acids, resin acids, lignans and oligomeric lignans, wherein lignans comprise at least following lignans: 7- hydroxymatairesinol, conidendrin, lariciresinol, liovil, secoisolariciresinol or their geometric isomers or stereoisomers thereof and wherein the amount of oligomeric lignans is 10 - 50 % of the total weight of said lignan mixture,

-mixing obtained alcohol-wood extract or an alcohol-water-wood extract, to a grinded meat of a hamburger, roll or like.

Example 6C

A salad dressing The compostion of the present invention may be added into any salad dressing by adding alcohol-wood extracts mixed with a suitable food grade composition or food additives mentioned before (see for example, multipurpose additives (subpart I) of CFR Code of Federal Regulations Title 21 .

Example 6D

Caramels

A part of sugar in making caramels may be replaced with the composition of the invention, comprising at a range of 1-100 wt-% preferably at a range of 5- 90 wt-% of lignan mixture. This composition can be an alcohol extract as pre- sented in table I as such or it can be an alcohol composition or water-alcohol based raw alcohol-wood extracts or alcohol-wood extracts mixed with a suita- ble food grade composition or food additives mentioned before (see for example, multipurpose additives (subpart I) of CFR Code of Federal Regulations Title 21 . The composition of the invention can be added before or after heating the caramel ingredients. Example 6E

A pill

A pill which can be a tablet or a capsule can be made from a food additive, dietary supplement or nutraceutical in a conventional way. For example, when making tablets, the composition of the invention can be added into a food addi- tive, a dietary supplement or a nutraceutical which is then powdered or granulated. This powder or granulate and a suitable binder such as sucrose, lactose, cellulose are mixed together and tablets are made from this mixture by pressing.

Claims

1 . The use of a composition comprising at a range of 1-50 wt-% preferably at a range of 5-50 wt-% a lignan mixture comprising fatty acids, resin acids, lignans and oligomeric lignans, wherein lignans comprise at least following lignans: 7-hydroxymatairesinol, conidendrin, lariciresinol, liovil, secoisolar- iciresinol or their geometric isomers or stereoisomers thereof and wherein the amount of oligomeric lignans is 10 - 50 % of the total weight of said lignan mixture, for lowering the cholesterol level of blood serum of a mammal.

2. The use of a composition defined in claim 1 wherein the oligomeric lignans are oligolignans comprising at least of sesterlignans, sesquilignans, and dilignans.

3. The use of a composition defined in claim 1 said composition additionally containing sterols such as sitosterols or sitostanols.

4. . The use of a compostion defined in claim 1 wherein lignan mixture also contains pinosylvin resin acid.

5. The use of a composition in claim 1 wherein lignan mixture also contains pinoresinol.

6. The use of a composition mentioned in claim 1 wherein lignan mixture comprises also one or more lignan(s) at a range of 0.1-10 wt-% selected from the group composing of: pinoresinol, nortrachelogenin, matairesinol, juva- biones, or their geometric isomers or stereoisomers thereof and oligolignans for lowering cholesterol level in a mammal.

7. The use of composition defined in claim 1 or 2 for lowering cholesterol level in a mammal by preventing inflammation of blood vessels.

8. The use of composition defined in claim 1 wherein the composition further comprises stilbenes or triterpens selected from the group composed of betulin, betulonic acid, betulinic acid, betuloinic acid, resveratrol and their geometric isomers or stereoisomers thereof.

9. The use of a composition defined in claim 1 wherein the composition fur- ther comprises vitamin or plant lignan(s).

10. The method of claim 1 , wherein said method additionally contains a step of measuring the cholesterol level of blood serum of a mammal and then administering cholesterol level lowering composition.

1 1 . The method according to claim 10 wherein the amount of composition for lowering the cholesterol level of blood serum is defined on the basis of results of cholesterol level measurement.

12. A method for producing a composition for lowering cholesterol level of blood serum in a mammal, wherein said composition is prepared by

- extracting knot wood and/or bark with an extraction solution selected from the group composed of alcohol and alcohol-water mixture,

- obtaining an alcohol-wood extract or an alcohol-water-wood extract, and

- adding a possible edible carrier agent to said alcohol-wood extract or alcohol- water-wood extract wherein said composition comprises at a range of 1-100 wt-% preferably at a range of 5-90 wt-% of lignan mixture and an extraction solution wherein said lignan mixture in extraction solution comprises fatty acids, resin acids, liovil, lignans and oligomeric lignans, wherein lignans comprise at least following lignans: 7-hydroxymatairesinol, conidendrin, lariciresinol, secoisolariciresinol or their geometric isomers or stereoisomers thereof and wherein the amount of oligomeric lignans or their geometric isomers or stereoisomers thereof is 10 - 50 % of the total weight of said lignan mixture present in said composition, wherein the extraction solution is selected from the group consisting of alcohol and water-alcohol mixture, the balance being a physiologically acceptable carrier agent.

13. The method defined in claim 12 comprising at a range of 1-80 wt-% preferably at a range of 5-80 wt-% of a lignan mixture comprising at least the following lignans: 7-hydroxymatairesinol, conidendrin, lariciresinol, liovil and secoisolariciresinol or their geometric isomers or stereoisomers and oligolignans which are oligomeric lignans comprising at least sesterlignans, sesquilignans, and dilignans.

14. The method defined in claim 12 wherein said composition comprises at a range of 5-50 wt-% preferably of 1-10 wt-% lignan mixture.

15. The method defined in any of the claims 12 - 14 wherein lignan mixture comprises also one or more lignan(s) at a range of 0.1-10 wt-% selected from the group composing of: pinoresinol, nortrachelogenin, matairesinol, juva- biones and oligolignans and their geometric isomers or stereoisomers thereof.

16. The method defined in any of the claims 12 - 15, wherein the extraction solution comprises alcohol which is a lower monovalent alcohol, a lower bivalent alkyl alcohol or a lower trivalent alcohol or mixture thereof.

17. The method defined in claim 16 wherein divalent alcohol is selected from the group composing of propylene glycol, butylene glycol and pentylene glycol and trivalent alcohol is glycerine.

18. The method defined in claim 12 wherein the extraction solution comprises monovalent lower alcohol such as ethanol and/or, divalent lower alcohol such as propanediol and/or trivalent lower alcohol such as propanetriol.

19. The method defined in claim 12 wherein knot wood originates from coniferous trees, selected from genus Picea, Abies or Tsuga.

20. The method defined in claim 12 wherein the knot wood originates from Picea abies, hemlock (Tsuga sp.) or from Acasia trees.

21 . The method defined in claim 12, wherein the concentration of said lignan mixture in prepared composition is following:

- 7-hydroxymatairesinol 70-80 %;

- conidendrin 3-8 %;

- lariciresinol 1-4 %; - liovil 2-5 %;

- secoisolarisinol 3-7 %;

- oligomeric lignans 0-3 %.

22. The method defined in claim 12 wherein the composition further comprises triterpens or stilbenes selected from the group composing of betulin, betulonic acid, betulinic acid, betuloinic acid and resveratrol or their geometric isomers or stereoisomers thereof.

23. The method defined in claim 12 wherein the composition further comprises vitamin(s) or plant lignan(s).

24. A composition prepared according to method defined in any of the previous claims 12-23.

25. A method for preparing a food additive, a dietary supplement or a nutraceutical for lowering cholesterol level of blood serum in a mammal, wherein said method comprises the following steps:

- extracting knot wood and/or bark of wood with an extraction solution selected from the group composed of alcohol and alcohol-water mixture for obtaining an alcohol-wood extract or an alcohol-water-wood extract, - adding a possible edible solid or liquid carrier agent such as oil or aquous liquid or emulsion to said alcohol-wood extract or alcohol -water-wood extract and

-mixing said alcohol-wood extract or alcohol-water-wood extract, in combination with a possible edible carrier agent to a food additive, dietary supplement or nutraceutical to prepare an edible mixture which is a food additive, a dietary supplement or a nutraceutical comprising at a range of 1-50 wt-% preferably at a range of 5-50 wt-% a lignan mixture comprising fatty acids, resin acids, lignans and oligomeric lignans, wherein lignans comprise at least following lignans: 7-hydroxymatairesinol, conidendrin, lariciresinol, liovil, secoisolar- iciresinol or their geometric isomers or stereoisomers thereof and wherein the amount of oligomeric lignans is 10 - 50 % of the total weight of said lignan mixture.

26. The method defined in claim 25 wherein the oligomeric lignans contain at least sesterlignans, sesquilignans and dilignans.

27. The method defined in claim 22 or 25 for preparing food additive dietary supplement or nutraceutical for lowering cholesterol level of blood serum in a mammal, which comprises at a range of 1-90 wt-% preferably at a range of 5- 90 wt-% said lignan mixture in a combination with an extraction solution se- lected from the group composing of alcohol and water-alcohol mixture the balance being an edible solid or a liquid carrier material.

28. The method defined in any of the claims 25 -27 wherein the food additive, dietary supplement or nutraceutical wherein said lignan mixture comprises also one or more lignan(s) at a range of 0-10 wt-% selected from the group composing of: pinoresinol, nortrachelogenin, matairesinol, juvabiones and oli- gomeric lignans, and their geometric isomers or stereoisomers thereof.

29. The method defined in any of the claims 25 -28 comprising step of adding to said food additive, dietary supplement or nutraceutical, stilbenes or triterpens selected from the group composed of betulin, betulonic acid, betulin- ic acid, betuloinic acid, resveratrol and their geometric isomers or stereoisomers thereof.

30. The method defined in any of the claims 25 -29 comprising step of adding to said food additive dietary supplement or nutraceutical vitamin or plant lignan(s).

31 . The method according to claim 25 wherein the method further comprises a step of forming a pill or a capsule from said food additive, dietary supplement or nutraceutical.

32. The pill or capsule prepared according to claim 31 and containing a food additive, dietary supplement or nutraceutical in combination with wood lignans.

33. The method defined in claim 25 further comprising a step of extending the shelf life of a food material by treating said food material with a food additive having antioxidative or antimicrobial effect.

34. The method defined in claim 32 further comprising a step of enhancing production of enterolactones in human gut, by administering orally the dietary supplement to a human being on daily basis.

35. The method defined in claim 25 further comprising a step of decreasing risk of cancer in a mammal by administering orally the dietary supplement to a human on daily basis.

36. A method of claim 25 further comprising making a fortified food product from said food additive, dietary supplement or nutraceutical of claim 22.

37. The method defined in claim 36 wherein the fortified food product is emulsion or liquid such as oil, sauce, dressing, yoghurt, juice, nectar or drink.

38. The method defined in claim 36 wherein the product is solid such us rice, pasta, macaroni or meat.