CN101316603A - Process for obtaining lignan extracts and compositions containing lignan extracts - Google Patents

Abstract

Methods for obtaining lignan extracts from plant material are disclosed. Compositions comprising lignan extracts and uses of lignan extracts are also disclosed.

Description

Process for obtaining lignan extracts and compositions containing lignan extracts

technical field

The present invention relates to a method for obtaining lignan extracts from plant material. The present invention also relates to compositions comprising lignan extracts obtained from the methods described herein and methods of use of compositions comprising lignan extracts.

Background technique

Lignans are secondary plant metabolites produced from shikimate via the phenylpropanoid pathway. They are derived from flavonoid precursors and are responsible for helping plants defend against pathogenic bacteria and predatory natural enemies. Lignans are defined as compounds having the structure of 2,3-benzhydrylbutane, and include pogrosinol, secoisolaricresinol, larchresinol, isolaricisinol, nordihydrocure Tranic acid, pinoresinol, oleoresin and other compounds, and variants thereof, including di(gluco)glucosides.

Flaxseed (cultivated flax) is probably the richest source of the phytoestrogens lignans. The primary lignan found in flaxseed is 2,3-bis(3-methoxy-4-hydroxybenzyl)butane-1,4 that is stored in the plant as a conjugated flax lignan (SDG) in the natural state - Diol (secoisolaricoresinol). Flaxseed contains 75-800 times more of these phytoestrogens than any other plant food. The plant lignan, catechin nordihydroguaiaretic acid, is a potent antioxidant previously used in the food industry.

Plant phenolic compounds occur as free monomers or in combination with other plant compounds to form esters or glucosides. Phenolic acids are known to have antioxidant activity. The major phenolic constituents in flaxseed were reported to be coumaric acid (4-glucosyl-cinnamic acid), caffeic acid (3-hydroxy-4-glucosyl-cinnamic acid), ferulic acid (3-methoxy -4-glucosyl-cinnamic acid) and hydroxymethylglutaric acid. These compounds are antioxidant and hypercholesterolemic.

Numerous reports in the literature have demonstrated the phytochemical benefits of flaxseed lignans. Rickard et al. reported that the supply of purified lignans at a dietary level of 5% flaxseed significantly reduced colon and mammary carcinogenesis in animals (Proceedings of the 57 th Flax Institute of the United States, (Fargo, N.D.): 8-13 (1998) ). Demark-Wahnefried et al. also reported that flaxseed supplementation can have beneficial effects on prostate cancer biology (Demark-Wahnefried et al., Adult Urology 58(1):47-52 (2001)).

In addition, lignans have been reported to prevent the development of type I and type II diabetes by 71% (Prasad, K-Proc. of the American Diabetes Association, (1999)), lignans as hypotensive agents can lower blood pressure without Affecting heart rate (US Patent No. 6,498,145), lignans are effective against lupus nephritis (US Patent No. 5,827,256), and lignans reduce the development of atherosclerosis caused by hypercholesterolemia in animals (Atherosclerosis 132:69- 76 (1997)), there are also many reports on possible antioxidants (Mol. & Cell. Biochem., 202: 91-100 (1999)) and anticancer properties (Anticancer Research 18: 1405-1408 (1998)).

Flaxseed in whole ground or defatted form has been incorporated into animal feed and foods such as bread, cookies, bagels and muffins. It is also used to boost fiber levels in meat products (WO00/19842). However, the available amount is limited because the high oil content of flax and the presence of plant mucilage lead to excess caloric intake and laxation (WO 96/30468).

Another issue associated with the use of flax in food is the toxicity of the cyanogenic glycosides present in flaxseed. Cyanogenic glycosides are nitrogen-containing secondary plant metabolites which, if overconsumed over a long period of time, can lead to goiter problems, damage to human organs, cyanide toxicity or possible thiocyanate effects on the thyroid gland. These glucosides are important natural toxins in animal and human nutrition. They have properties specific to flaxseed to help animals resist the toxic effects of ingesting selenium (Smith, et al., J. Org. Chem., 45:507-510 (1980)). The main cyanogenic glycosides present in flaxseed are linustatin, neolinusstatin and linamarin, and linustatin accounts for 54-76% of the total cyanogenic glycosides. Defatting flaxseed meal with hexane is known to produce an enrichment of all individual cyanogenic glycosides on a dietary isobaric basis (Maaza and Oomah, in Flaxseed in Human Nutrition, Cunnane and Thomson eds. AOCSPress, Champaign, III.1995 ). Therefore, it is crucial to separate cyanogenic glycosides from other compounds present in flax.

The literature has reported methods for the preparation of lignans and other phenolic compounds. In 1956, Bakke and Klosterman described a method for extracting lignans from defatted flax using methanolic dioxane (Proceedings of the North Dakota Academy of Sciences 10:18-22 (1956)). However, lignans are known to occur in flax as "complexes" with cinnamic acid glycosides and other compounds. Thus, originally lignans meant "polymers" in flax. Sodium methoxide and barium methoxide are used in methanolysis to separate lignans from other compounds (Blake and Klosterman; Thompson et al., Nutr. Cancer 26:59-165 (1996)). Almost all lignans present in flaxseed occur as components of soluble ester-linked complexes and not as free glycosides or aglycones (Muir et al., Proc. of the 58 th Flax Institute of the US, Fargo, N.D., 1999). A detailed review of various methods of extracting lignans and cinnamic acids can be found in Muir, supra. U.S. Patent No. 5,705,618 and PCT applications WO 96/30468 and WO 00/78771 also describe the preparation of lignan-containing complexes. method. However, these methods are deficient because they cannot be scaled up commercially due to the difficulty in achieving the required separation and purification without complex solvent systems such as ethyl acetate/water, and difficult to use chromatographic techniques or expensive methods such as size exclusion chromatography. US Patent No. 6,767,565 describes a method of purifying lignans using membrane and resin chromatography. However, this method uses expensive resins and membranes that are poorly stable in ethanol.

Thus, there is a need for more efficient methods of isolating lignans and compositions comprising those lignans.

Summary of the invention

In one embodiment, a composition comprises a lignan extract comprising 20-45% flax lignans. The composition also includes 50 mg or less of cyanogenic glycosides per 100 g of composition.

In another embodiment, a composition comprising a first lignan extract having less than 50% flax lignans and a second lignan extract having more than 50% flax lignans .

In a further embodiment, a method of producing a lignan extract comprises contacting a lignan-containing plant material with an extraction solvent, thereby forming an extraction solution. The method also includes separating the lignan-rich fraction from the extraction solution using a density-based separation device and washing the lignan-rich fraction with an aqueous solution. The method may further comprise drying the lignan-rich fraction.

Also disclosed is the use of the composition or the lignan extract for lowering cholesterol levels, treating benign prostatic hypertrophy, treating heart disease, preventing hypercholesterolemic arteriosclerosis, alleviating heart disease, lowering serum cholesterol, and reducing ischemic damage , enhance the expression of nitric oxide in endothelial cells, inhibit the activity of cyclooxygenase in macrophages, reduce the symptoms of diabetes, prevent or delay the onset of diabetes, adjust serum glucose levels, increase bone density, reduce bone density loss, relieve arthritis symptoms, reduce Use for prostate cancer risk, reduction of risk of metastasis, delay of metastasis, reduction of menstrual onset symptoms, reduction of menopausal onset symptoms, reduction of blood pressure, or reduction of hair loss or male pattern baldness. The use of the compositions or extracts for the preparation of medicaments for the treatment of these conditions is further described.

Further aspects of the invention relate to compositions and formulations comprising the lignan fraction obtained by the above method and to the use of these compositions and formulations.

Description of drawings

Figures 1A to 1D are molecular weight distributions extracted from lignans produced using the method of the present invention.

Figure 2 shows a flow chart illustrating the steps of one embodiment of the method of extracting lignans of the present invention.

Figures 3-4 are graphs illustrating the total cholesterol (TC) lowering capacity of one embodiment of a lignan extract of the present invention.

Figures 5-6 are graphs illustrating the low density lipoprotein cholesterol (LDLC) lowering capacity of one embodiment of a lignan extract of the present invention.

7-8 are graphs illustrating the high-density lipoprotein cholesterol (HDLC) raising ability of one embodiment of a lignan extract of the present invention.

Figures 9-10 are graphs illustrating the triglyceride (TG) reducing capacity of one embodiment of a lignan extract of the present invention.

Figure 11 is a graph showing plasma levels of lignan metabolites in patients treated with one lignan extract embodiment of the present invention.

Detailed description of the invention

Research on isolating or purifying lignan extracts continues. US Patent 6,767,565, assigned to Decatur, Illinois, Archer Daniels Midland Company, the entire contents of which are incorporated herein by reference, describes the use of membranes and resins to obtain lignan extracts from flaxseed. The present invention relates to an improved method for the extraction of isolated or purified lignans. In various embodiments, the invention discloses methods of obtaining lignan extracts and compositions containing lignan extracts and uses thereof.

In one embodiment, a method may comprise contacting lignan-containing plant material with an extraction solvent to obtain an extraction solution comprising a lignan-containing extract and an extraction solvent, separating plant material solids from said extraction solution, The content of microbial components in the extraction solution is reduced, 0-100% of the extraction solvent is removed from the extraction solution, and an aqueous diluent is added to the extraction solution, from which lignans are obtained.

In another embodiment, a method of obtaining lignans may comprise contacting a lignan-containing plant material with an extraction solvent to obtain an extraction solution comprising a lignan-containing extract and an extraction solvent, separating Plant-derived solids, reducing the content of microbial components in the extraction solution, contacting the extraction solution with an adsorption resin, washing the adsorbed lignan-containing extract with an aqueous diluent, and eluting the lignan-containing extract, thus obtaining a lignan-containing extract phenolic compounds, and optionally adjust the pH of the released lignans prior to drying.

In a further embodiment, a process comprises a method of obtaining lignans from plant material comprising: contacting the lignan-containing plant material with an extraction solvent to obtain an extraction comprising the lignan-containing extract and the extraction solvent solution, separating plant material solids from said extraction solution, reducing the content of microbial components in the extraction solution, removing 0-100% of the extraction solvent from the extraction solution, and simultaneously adding an aqueous diluent, wherein lignans was obtained from it.

In one embodiment, plant material such as flax, flaxseed or defatted flax may be comminuted by any suitable method such as grinding, crushing and/or chopping. The comminuted plant material can be extracted with a suitable extraction solvent such as alcohol. Alcohols may be aqueous alcohols or aqueous fatty alcohols. Suitable alcohols may be selected from methanol, ethanol, isopropanol or any combination thereof. In another embodiment, an aqueous solution of acetone may be used. The alcohol concentration should be in the range of 50-80% by volume as described by Westcott and Muir, "Medicinal Lignans from Flax Seed", Phytochemicals and Phyto-pharmaceuticals, AOCS Press, Champaign, III., 2000, and may be 60-70% by volume.

In certain embodiments where the raw material used is defatted flaxseed meal, the comminuted material can be screened and aspirated to remove fines produced by the process and larger particles of fines which will fall when placed into the extractor . This fines can cause problems in some subsequent extraction steps. Screening can be accomplished using a combination of two sieves with different pore sizes for producing lignan extracts with an intermediate section. Screening is the removal of coarse material which may contain litter, agglomerated fines and fatty pieces of soap stock in defatted flaxseed meal. In one embodiment, a first sieve of 600 microns may be used in combination with a second sieve of 2500 microns for screening defatted flaxseed meal. The portion of flaxseed meal retained between the two sieves can be used to produce lignan extract. A fine mesh is also used to remove fine particles that clog the extractor screen and reduce the percolation rate. The middle sieve knife can be sucked off and the "light" fraction collected. The "light" sections in the aspirator are typically the sections that offer more resistance to air movement and thus create airflow. Sometimes double suction is also used. The SDG of irregularly shaped husk flakes aggregated in the light fraction is higher than the thicker heavy particles that also offer less resistance to airflow and are removed as the "heavy" fraction. The irregular shape of the shell pieces also prevents the extractor filter residue from packing together. The product of this process is well filtered in liquid-solid extractors (such as those available from Crown Ironworks, Minneapolis MN) without clogging the screens used to drain the extraction solvent. The suction conditions are readily determined by those skilled in the art and should ensure removal of the less dense/higher air resistance fraction from the heavier/lower air resistance particles.

In other certain embodiments, the plant material used to obtain the lignan extract may be selected from: pumpkin seeds, fiber-rich plants such as grains including but not limited to wheat, barley or oats, legumes such as kidney beans, lentils or soybeans, vegetables such as garlic, asparagus, cauliflower or carrots.

Extraction may be performed at a suitable temperature to minimize the growth of potentially harmful microbial compounds and facilitate extraction of the lignan complex from the plant material. Suitable temperatures to minimize microbial growth are between room temperature and the boiling point of the solvent, such as between 40°C and 70°C or between 50°C and 60°C.

The ratio of extraction solvent to comminuted plant material should be suitable for agitating the mixture. This ratio may be from 1:1 to 8:1 ml solvent/gram comminuted plant material. Another ratio is 2:1 to 6:1 ml solvent/gram comminuted plant material.

The duration of the extraction process is about 4-8 hours. In another embodiment, extraction may be performed for about 5 to 7 hours. When using high temperature, the extraction time can be 6 hours. In another embodiment, the method can be performed in a continuous mode of current mode or reverse current mode as understood by those skilled in the art. The residence time to obtain an optimum extract from flaxseed meal can vary. This time can be determined by the time, temperature and ratio of extraction solvent to comminuted plant material. The extraction time can vary from 0.1 hour to 10 hours. In another embodiment, a residence time of 0.75 hours can be used in the reverse current extractor.

This extraction method results in the formation of a slurry comprising an extraction solvent containing the desired lignan components and solid plant material. After completion of the extraction process, the resulting liquid portion of the slurry (hereinafter referred to as extraction solution), which contains the desired lignan complexes, can be separated from the solid plant material. Separation can be carried out in any suitable manner known to remove solids from liquids. Examples of suitable techniques include, but are not limited to, filtration, sedimentation, sedimentation, standing, centrifugation, and any combination thereof. When filtration is used, the pore or mesh size of the membrane, filter, or sieve is selected according to the size of the material to be removed. The type of membrane filter is selected based on the compatibility of the membrane filter with the solvent used in the extraction method.

When the plant material was flax, it was observed that the flax swelled during extraction and subsequently absorbed a large amount of solvent at the end of the extraction. The slurry thus forms a wet cake which can be strained and/or pressed to remove the hydroalcoholic extract from the solid plant material components. Thus, in one embodiment, the wet cake may be pressed, such as with a screw press, to obtain an extraction solution. This greatly increased the yield of lignan complexes from flax meal.

To further increase the yield of the extraction solution, a screw press can also be used after the separation of the plant material solids and the extraction solution, wherein eg filtration, sedimentation, sedimentation, standing and/or centrifugation are used as separation techniques.

The resulting extraction solution can be further purified by passing it through a membrane filter to reduce the level of microbial components in the solution and/or remove any residual solids not removed from the initial separation. One type of membrane filter used for this purification may be a nylon filter, however, any suitable type of filter may be used. In one embodiment, the percent reduction in microbial content is between about 90-100%, 90%, 95%, possibly about 98-100%, and possibly 99.9%.

Nylon membrane filters may have a pore size of 0.1-10 μm or 0.2-10 μm. Membrane cartridge filters with similar pore size values may also be used. The resulting clear extraction solution can be further processed according to, but not limited to, one embodiment described below.

The alcoholic extraction solvent can be removed (ie, stripped) from the clear extraction solution, such as by evaporation. It was observed that concentration of the extraction solution by evaporation resulted in the formation of a thick sticky layer that rapidly coalesced as a precipitate.

In order to avoid the effect of evaporating the extraction solvent and to obtain a good treatment of the extract, the extraction solution can be diluted by adding an aqueous diluent during the evaporation/stripping of the extraction solvent. In one embodiment, the aqueous diluent may be water. Thus, most of the extraction solvent in the extraction solution can be removed if desired. In this way, water can be added to maintain the desired dilution of the extract during the evaporation/stripping step.

In one embodiment, the resulting aqueous extract solution (ie, the supernatant aqueous layer) may be dried to obtain a powder. Drying can be performed by any suitable method including, but not limited to, evaporation, vacuum drying, freeze drying, spray drying, or any combination thereof.

In another embodiment, after reducing the alcohol content of the clarified extraction solution by dilution and/or evaporation in order to obtain a less hygroscopic product, the resulting extraction solution may be subjected to ultrafiltration to remove sugars present therein (for example, raw cyanoglycosides). Ultrafiltration can be performed using at least two filter volumes of aqueous diluent to remove cyanogenic glycosides and other unwanted compounds. In one embodiment, the aqueous diluent may be water.

Suitable ultrafiltration membrane filters include organic polymers or copolymers such as polysulfone, polyacrylonitrile, cellulose acetate, or inorganic materials such as zirconia, alumina or ceramic materials. Depending on the molecular weight distribution of the desired lignan product, a membrane filter with a molecular weight cutoff (MWCO) of less than 5,000 can be used; or in another embodiment, a membrane filter with a MWCO of 1,000 can be used.

Due to the poor stability of polymer films in high-concentration alcohol solutions (Shukla, R.Ph.D. Thesis, University of Illinois at Urbana-Champaign, 2000), the alcohol concentration can also be reduced by diluting with water or stripping alcohol such as evaporation. Thus, the resulting reduced alcohol content extraction solution has no negative impact on the operation of the membrane filter.

The resulting ultrafiltration retentate is substantially free of cyanogenic sugars and contains the desired lignan extract in relatively purified form. The ultrafiltration retentate can be dried directly or after further concentration by evaporation by one or more of the following techniques: vacuum dryer, microwave dryer, radio frequency dryer, and/or freeze dryer. For example, vacuum drying can be accomplished at an absolute pressure of 5-20 mbar, a residence time of 25-40 minutes, and an evaporation temperature of 95-115 degrees Fahrenheit. Drying under these conditions can be accomplished using a vacuum belt dryer.

The vacuum belt dryer consists of a frame with a built-in conveyor belt for transporting the product to be dried onto a heating plate. Automatic belt control ensures precise belt operation. Such a dryer is available from Merk Process (Laufenberg, Germany). Bands are parallel to each other and metering pumps with vibrating application nozzles are assigned to each individual control band. The change in boiling point under vacuum provides a low evaporation temperature and thus a mild product temperature. Pumpable products are usually in a highly viscous, often viscous phase during the drying process which can lead to the formation of vapor bubbles in the product and dry residue left on the belt at the end of the drying process. The temperature given to the product during the drying process has a significant effect on product quality. The belt runs through a number of heating zones and at the end through a cooling zone. Steam, pressurized water or thermal oil can be used for heating and each zone can be controlled individually to allow the heating surface temperature to be adjusted to the specific requirements of the product being dried. Utilization of such a dryer reduces loss of product quality and formation of unwanted coloration, and provides a free-flowing shelf-stable product.

In another embodiment, the extraction solution can be diluted with an aqueous diluent, and instead of removing the extraction solvent, the diluted extraction solution can be subjected to ultrafiltration to remove sugars and other impurities.

In another embodiment, the invention discloses a method of obtaining a lignan extract comprising: contacting a lignan-containing plant material with an extraction solvent to obtain an extraction solution comprising a lignan-containing extract and an extraction solvent, from Separating the plant-derived solids from the extraction solution, reducing the content of microbial components in the extraction solution, contacting the extraction solution with the adsorption resin, washing the adsorbed lignan-containing extract with an aqueous diluent, and eluting the lignan-containing extract , thereby obtaining lignans, and optionally adjusting the pH of the released lignans before drying.

In another embodiment, the clarified extract solution can be treated with an adsorption resin batch or continuous column chromatography type. The extract is applied to an adsorbent resin and washed with an aqueous diluent to remove cyanogenic sugars and other impurities. The aqueous diluent can be water.

The lignan complexes adsorbed on the resin can be treated with aqueous alcohol (20-100% by volume at 25-85°C) gradient or single percentage dilution. The alcohol may be selected from, but not limited to, methanol, ethanol, isopropanol or any combination thereof. The resulting material can be dried by, for example, evaporation, vacuum drying, freeze drying, spray drying, or any combination thereof to yield a product of about 20-45% by weight lignans. It has been found that adjusting the pH of the chromatographically separated product does not destroy the lignan complexes, but facilitates drying of the product. The pH can be adjusted with any suitable acidic or basic agent. In one embodiment, adjusting the pH may include adjusting the pH of the released lignan complex to between about 3.0 and about 9.0 prior to drying the product. In another embodiment, the pH can be adjusted to between about 7.5 and about 8.0.

Suitable resins that may be used are selected from, but not limited to, polymethacrylates, ethylvinylbenzene (ethylvinylbenzene)-divinylbenzene, styrene-divinylbenzene, polystyrene or phenol formaldehyde polymers, and Can be ionic or nonionic.

In another embodiment, the extraction solution may be diluted with an aqueous diluent prior to treatment with the adsorbent resin. The aqueous diluent can be water.

In another embodiment, the pH of the extraction solution can be adjusted to a value between about 3.0 and about 9.0. In one embodiment, the pH is between about 7.5 and about 8.0. It has been found that adjusting the pH with any suitable acidic or basic agent does not destroy the lignan complexes, but rather facilitates the subsequent drying of the lignan product.

In another embodiment, a lignan extract is produced as described in the flow diagram of FIG. 2 . The defatted flaxseed meal was obtained by deoiling crushed flaxseed (Linum uistatissimum) using n-hexane extraction. The desolventized defatted flaxseed meal can optionally be mechanically (ie, physically) sorted to an optimal particle size/shape. This sorting can be accomplished by a combination of screening and aspiration as described herein. The desolvated, and optionally sorted, defatted flaxseed meal may be mixed with aqueous ethanol at 45-75° C. at 60-95% ethanol by volume. The ethanol/linseed meal mixture can be filtered to remove any insolubles produced in the ethanol soluble extract.

Ethanol soluble extracts can be evaporated under vacuum to substantially remove all ethanol. Depending on the temperature at which evaporation occurs, the vacuum can be adjusted to remove ethanol. Evaporated soluble extracts containing lignan complexes can be separated from soluble and suspendable soluble impurities. In one embodiment, the lignan complex can be separated from soluble and suspendable soluble impurities (i.e., non-lignans) using a density-based separation device or a centrifuge device (e.g., a hydrocyclone or a continuous centrifuge). substances) to coalesce and/or deposit a viscous liquid rich in lignan complexes, which can be decanted from the supernatant separated from the centrifuge. This method may also be called deposition coalescence.

In another embodiment, the lignan complexes may be separated from soluble and suspendable soluble impurities by precipitating, sedimenting, or agglomerating the lignan complexes. The use of settling or settling tanks is more economical than centrifuge or hydrocyclone type systems because no mechanical parts are involved. Furthermore, the simplicity of precipitation, deposition or coalescence provides a lower cost basis for manufacturing the product. Precipitation, settling or coalescence may be effected by pumping the evaporated soluble extract into a tank and allowing the lignan complex to settle, coalesce or fall to the bottom of the tank. In other embodiments, the precipitation, deposition or coalescence of the lignan complexes can be aided by adjusting the pH. In another embodiment, a filter may be used to separate the lignan complex from soluble and suspendable soluble impurities. Any such filter known in the art may be used including, but not limited to, polymeric or inorganic filters. The pore size of such filters can range from 0.01 microns to 100 microns.

The lignan-containing process intermediate may be rinsed by contacting the process intermediate with an aqueous citrate buffer solution. Flushing process intermediates removes cyanogenic glycosides from the lignan-rich complexes (ie, process intermediates). The citrate buffer may comprise a 0.1% citrate buffer at a pH of about 5, or up to about 10% citrate buffer. The pH can be varied within an acidic range such as pH 3.0 to 6.0 to maximize removal of cyanogenic glycosides and minimize loss of soluble lignans. In other embodiments, rinsing of the lignan complex may also be performed with water at neutral pH, or alternatively with chelating agents including, but not limited to, ethylenediaminetetraacetic acid salt (ie, EDTA).

The rinsed lignan-rich complex is dried, optionally under vacuum, with or without preheating. The dried lignan-rich complex can be ground mechanically (i.e. using a Fitz mill, hammer mill or other similar equipment) to a specified size and can be packaged in polyethylene film, polyethylene drums for transport, placed in in food, or in capsules or tablets. In one embodiment, the lignan-rich complexes may be comminuted such that at least 90% of the lignan-rich complexes are 425 μm (micrometers) or smaller. Evaporated ethanol can be recycled or reused without rectification by combining concentrated vapors from ethanol evaporation and adjusting the ethanol solution proof if necessary.

In another embodiment, the lignan extract may be shaped for oral administration to a patient, eg, in the form of a food additive, beverage additive, tablet, capsule, multi-ingredient nutritional supplement, or any combination thereof. For example, lignan extracts can be added to flaxseed oil supplements, fish oil supplements, omega-3 fatty acid supplements, or other dietary supplements for human use. The lignan extract can be used directly as a food additive or mixed with a consumable carrier as a food additive, food composition, beverage additive or beverage composition. Non-limiting examples of food products include, but are not limited to, nutritional bars, cereals, snacks, wafers, biscuits, cookies, baked goods, dairy products, processed foods in blocks, nougat, or any other food product . The lignan extracts may also be incorporated into or mixed with consumable foods such as food compositions for human use, livestock or animal foods.

It is also desirable to make lignan extracts into capsules, tablets, caplets or liquid forms for routine administration to help treat and lower cholesterol levels, treat benign prostatic hypertrophy, treat heart disease, and prevent hypercholesterolemic arteriosclerosis , Relieve heart disease, reduce serum cholesterol, reduce ischemic damage, enhance nitric oxide expression in endothelial cells, inhibit cyclooxygenase activity in macrophages, relieve diabetes symptoms, prevent or delay the onset of diabetes, adjust serum glucose levels, increase Bone density, decrease loss of bone density, decrease symptoms of arthritis, decrease risk of prostate cancer, decrease risk of metastasis, delay metastasis, decrease symptoms of menstrual onset, decrease symptoms of menopausal onset, lower blood pressure, or reduce conditions associated with hair loss or male pattern baldness. Beverages enriched with lignan extracts may be prepared for a similar purpose, allowing individuals to take advantage of the known benefits of lignans. Beverages include, but are not limited to, apple juice, grape juice, cranberry juice, orange juice, energy drinks, sports drinks, meal replacement drinks, other fruit juices, any other soft drinks, and dairy products such as milk, alcoholic beverages, or any combination thereof .

In another embodiment, the lignan extract can be combined with any other compound or food additive such as sterols, isoflavones, sweeteners, protein isolates or concentrates, non-nutritive sweeteners, soluble or insoluble fibers, statins , vitamin E, proanthocyanidins, saw palmetto extract, pygeum extract, linseed oil, fish oil, vitamin B, vitamin C, tocopherol, phytosterols, polyphenols, catechins, anthocyanins, astaxanthin, glucose Glucosamines and combinations thereof in any combination.

In a further embodiment, the lignan extract produced by one of the methods of the present invention can be used in cosmetics such as cosmetic, dermal or epidermal effects in an effort to alter pigmentation. Cosmetics may be formulated for oral administration as described herein or for topical application such as lotions, sunscreens or other topical applications.

The following non-limiting examples further describe the invention. Those skilled in the art will appreciate that some possible changes of these embodiments are within the scope of the present invention and various steps in the method can be used in combination.

specific implementation plan

Example 1

Lignan extracts were produced as described in the flow diagram of Figure 2. The defatted flaxseed meal was obtained by deoiling crushed flaxseed (Linum uistatissimum) using n-hexane extraction. The desolventized defatted flaxseed meal can optionally be mechanically (ie, physically) sorted to the optimum particle size/shape described herein. The desolvated, and optionally sorted, defatted flaxseed meal may be mixed with aqueous ethanol at 45-75° C. at 60-95% ethanol by volume. The ethanol/linseed meal mixture can optionally be filtered to remove any insolubles, resulting in an ethanol soluble extract.

Ethanol soluble extracts can be evaporated under vacuum to substantially remove all ethanol. Precipitation or deposition may be performed by pumping the evaporated soluble extract into a tank and allowing the lignan complex to coalesce and/or settle or fall to the bottom of the tank. Optionally, this separation can also be accomplished using density-based separation equipment such as hydrocyclones or centrifuges. Process intermediates containing lignan complexes can be rinsed by contacting the process intermediates with aqueous citrate buffer (Figure 2). In an alternative embodiment, solutions containing organic acids such as acetic acid, fumaric acid, lactic acid, ascorbic acid may be used. The rinsed lignan-rich complex is dried, optionally under vacuum, with or without preheating, or optionally using a freeze dryer or vacuum belt dryer. Table 1 provides the analysis of the lignan extract produced in this example.

Lignan extracts obtained using the methods described herein include SDG or β-D-glucopyranoside, (2R,3R)-2,3-bis[(4-hydroxy-3-methoxyphenyl)]formazan ]-1,4 _- butanediylbis-(9Cl), having _a molecular formula of _C32H46O16 and a calculated molecular weight of about 686.7. Lignan extract is available in powder form and may be brown in color, "tea" like smell and taste, slightly soluble in water at neutral pH, moderately soluble in 70% ethanol, and has low hygroscopicity.

Five batches of lignan extracts were prepared using the method of this example. The 5 batches of lignan extracts had the following properties:

Lignan extracts produced using this example were also analyzed for cyanogenic glycoside levels using the methods described herein. A typical recommended human consumption of cyanogenic glycosides is less than about 50 mg/adult/day. As shown in Table 1, the lignan extract contained 50 mg or less of total cyanogenic glycosides/100 g of lignan extract.

Various lignan-rich extracts (ie, Lignan Extracts 1-6) were produced and analyzed using the method of Example 1. The various products analyzed were found to have the following properties:

Various lignan-rich extracts (i.e., Lignan Extract 7), total flaxseed, defatted flaxseed meal, sorted flaxseed meal and two process intermediates produced using the method of Example 1 were also do analysis. The various products analyzed were found to have the following properties as shown in Table 3:

All analyzed products were hydrolyzed prior to analysis and analyzed by HPLC for lignans and phenolic compounds. The hydrolysis step includes placing the product in 56% aqueous methanol containing 0.1M NaOH, heating at 65°C for one hour, and neutralizing it with dilute acetic acid. Other measurements were made using routine analytical methods.

Example 2

Once obtained from the methods described above, the lignan product can be added to food, beverage, nutritional or pharmaceutical formulations. The formulations can be formulated for any route of administration to humans or animals such as oral formulations. In one embodiment, the formulation comprising the resulting lignan extract may comprise 15-25 wt% lignan extract obtained by any of the methods described above, 60-84 wt% filler component, 1-25 wt% dietary supplement nutrient.

In other embodiments, the lignan extract placed in a food or other composition may include from about 7% to about 90% SDG, depending on the desired use of the lignan extract.

Example 3

Lignan extracts are placed in capsules or tablets for administration to patients. Capsules or tablets may be placed in a container such as a bottle with a label or instructions indicating to the user the recommended dose of lignan extract. In various embodiments, the recommended dose may be about 86-860 mg lignan extract/day, which is about 30-300 mg SDG/day; or in another embodiment, about 86-1720 mg lignan extract/day, which is About 30-600mgSDG/day. When such amounts of lignan extract are combined with excipients, fillers or other ingredients, the recommended dosage for the dietary supplement may be from about 250 mg to about 1.5 grams. Labeling or statements instructing users that the lignan extract contains low levels of cyanogenic glycosides, such as 50 mg or less total cyanogenic glycosides per 100 g of lignan extract, may also be used in conjunction with dietary supplements or dietary supplements containing container association.

While the various embodiments described herein are shown to contain certain concentrations of SDG, it will be apparent to those skilled in the art that the concentration of SDG can vary. For example, the lignan or SDG content of the plant material from which the lignan extract is analyzed can vary and be affected by many factors including, but not limited to, the species of plant material, the location of the plant material, the year the plant material was Extraction methods used to extract lignan extracts, or hydrolysis and analytical techniques used to measure SDG content. For example, it has been reported that the SDG content in flax products is between 29-1055mg/100g seed or 200-1300mg/100g flaxseed food. In addition, SDG contents in different varieties of flax have been reported to be in the range of 0.96-3.15 μmol/g. Thus, lignan extracts or compositions having any desired concentration of SDG in the range of about 7-89% can be obtained using the various embodiments described herein and incorporated into foods, nutritional compositions or pharmaceuticals using the methods described herein composition.

Example 4

In another embodiment, the effect of extracting lignans produced in Example 1 by centrifugation is studied in a clinical experiment. The clinical trial analyzed plasma lipids in hypercholesterolemic patients (20 females, 35 males, age 28-79 years) for 8 weeks using a randomized, placebo-controlled, double-blind design. Lignan extracts were formulated as dietary supplement tablets at the following SDG doses: 0 mg SDG (placebo), 300 mg SDG and 600 mg SDG per patient per day. As shown in Figures 3-6, treatment doses of 300 mg and 600 mg SDG significantly reduced total cholesterol (TC) and low-density lipoprotein cholesterol (LDLC) compared to placebo. High-density lipoprotein cholesterol (HDLC) was also reduced as shown in Figures 7-8. Triglyceride (TG) levels also decreased as shown in Figures 9-10. Plasma levels of lignan metabolites in patients are shown in FIG. 11 .

Clinical trials also showed that similar levels of creatine, blood urea nitrogen (BUN), alanine aminotransferase (ALT) and gamma-glutamyltransferase (GGT) were observed in the treatment groups and all these parameters were within the normal range. The data presented in Table 4 show that the lignan extract had no significant effect on kidney or liver function.

In this embodiment, the patient's plasma levels of intestinal lignans (ie, enterodiol and enterolactone) are also analyzed. Enterodiol plasma levels averaged 8.0 ng/ml for all patients in the control group and 229.8 ng/ml for all patients receiving 600 mg SDG during the supplementation period (28-fold difference), as described in Table 5. Plasma levels of intestinal lignans suggest that administering the lignans produced in Example 1 using centrifugation to extract and produce increased blood levels of intestinal lignans and enterodiols in human patients.

**Means excluding baseline levels show the effect of SDG supplementation compared to control.

Example 5

In a further embodiment, the effect of extracting the lignans produced in Example 1 by centrifugation was studied in a clinical experiment. A study was conducted to determine the effect of lignan extract on benign prostatic hyperplasia (BPH) and prostate specific antigen (PSA) in 60 men afflicted with BPH conditions. A randomized placebo-controlled double-blind design was used. The amount of lignan extract formulated into a dietary supplement provides 0 mg lignan extract (placebo), the SDG in 300 mg lignan extract, or the SDG in 600 mg lignan extract during 16 weeks. dose. Blood levels of creatine, blood urea nitrogen (BUN), alanine aminotransferase (ALT) and γ-glutamyltransferase (GGT) were measured at 0, 2 and 4 months. Based on the results shown in Table 6, administration of lignan extract in this study did not affect renal or hepatic function.

Significant relieving effects of flax lignans on certain BPH symptoms were observed. Blood levels of creatine, blood urea nitrogen (BUN), alanine aminotransferase (ALT) and γ-glutamyltransferase (GGT) were measured at 0, 8, and 16 weeks, and no significant differences between treatment groups were observed. difference. Based on the normal levels of biomarkers as shown in Table 7, the results of this study also showed that lignan extract had no negative effect on kidney or liver function.

Summary results for quality of life (QOL), International Prostate Symptom Score (IPSS), voiding blockage grade are presented in Tables 8-9. Compared with placebo, the 600mg SDG treatment group achieved a significant overall improvement in QOL scores (p=0.0115). Regarding the 300 mg SDG treatment group, although the QOL score improved, it did not reach statistical significance (p=0.1377). The percent change from baseline in the QOL score had statistically similar results to the QOL score itself (p=0.0085 and p=0.1248 for 600 mg SDG treatment and 300 mg SDG treatment compared to placebo, respectively). Significant treatment effects were seen in each group of patients. However, the SDG treatment group had a stronger effect on QOL score reduction and had a smaller p-value than the placebo group.

IPSS data were collected based on 7 questions related to voiding experience. Each question shows the voiding symptoms experienced by BPH patients. Each symptom is scored from 0 to 5. Higher scores represent more severe symptoms. Thus, the worst IPSS may be 35. In this study, IPSS was significantly reduced in all groups. Between treatment groups, an overall significant effect was not achieved compared with placebo. However, after 4 months of treatment, IPSS was significantly lower in the 300 mg SDG and 600 mg SDG treatment groups than in the placebo group (p=0.002 and p=0.018, respectively). The statistical results for percent change from baseline were similar to IPSS change. Regarding the data of the 3rd month and the 4th month, the percentage IPSS reduction effects of the 300mgSDG treatment group and the 600mgSDG treatment group were slightly significant (p=0.056 and p=0.081, respectively); while the data of the 4th month alone, both The reduction for both treatment groups was significant (p=0.0414 and p=0.0412, respectively). The noted significance is mainly due to the decrease in the IPSS-reducing effect in the placebo group over time, while the reduction in the SDG-treated group was maintained throughout the study period.

Within-group effects occurred in the placebo and treatment groups. Compared to baseline, IPSS was significantly lower at 2, 3, or 4 months. Table 8 presents IPSS percentage change data from baseline. Both treatment groups achieved a very significant intragroup reduction in IPSS, whereas the placebo group did not (Table 8).

The voiding blockage scale is used to assign the difficulty of urination in BPH patients. The Voiding Blockage Scale includes a set of scores for voiding symptoms identical to the IPSS set. The block rating also forms part of the IPSS. Table 9 shows that a significant treatment effect on block grade was obtained compared to placebo up to Month 4 with p=0.019 and p=0.012 for the 300 mg SDG and 600 mg SDG treatment groups, respectively. The achieved significance was primarily attributable to the return of the placebo-reducing effect on block grade to baseline while the effect of the treatment group was maintained. The within-group reduction effect on block grade was significantly lower than baseline in both the 300 mg SDG and 600 mg SDG groups at 2, 3, or 4 months (p>0.005). With respect to the placebo group, only the data for the 2nd and 3rd months reached significance (p<0.05). A significant treatment effect in reducing block grade was only observed at Month 4 (p=0.024 and p=0.013 for 300 mg SDG and 600 mg SDG compared to placebo, respectively). Both the 300 mg SDG treatment group and the 600 mg SDG treatment group strongly exhibited within-group treatment effects (p<0.005 for both monthly data). In the placebo group, only the data for the 3rd month reached significance (p<0.05).

Example 6

In another embodiment, the effect of extracting lignans produced in Example 1 by centrifugation is studied in a clinical experiment. A clinical trial directed study of the effects of phytosterols alone or in combination with lignan extracts on serum cholesterol in 25 men and 11 women over an 8-week period. A randomized placebo-controlled double-blind design was used to assign patients to the following treatment groups: placebo, 1.3 g sterols, 1.3 g sterols + 300 mg lignan extract. Blood levels of creatine, blood urea nitrogen (BUN), alanine aminotransferase (ALT) and γ-glutamyltransferase (GGT) were measured at 0, 2, 4, 6 and 8 weeks. In this embodiment, no differences in the concentrations of these biomarkers between and within treatment groups indicated that flax lignans had no negative impact on kidney or liver function, as shown in Table 10A. The data showed that the phytosterol-bound lignan extract lowered total cholesterol by 13.5% and lowered LDL-C by 18.9%, as shown in Tables 10B-10D.

^1All the above averages are within the normal range. ^* L = lignan extract.

^a = p <0.05; ^b = p < 0.01, compared to baseline, ^* L = lignan extract

^a = p <0.05; ^b = p < 0.01, compared to baseline, ^* L = lignan extract

^a = p <0.05; ^b = p < 0.01, compared to baseline, ^* L = lignan extract

Example 7

In a further embodiment, in order to maintain a constant amount of SDG in the lignan extract, the first amount of lignan extract, e.g. obtained using the method of the present invention, i.e. about 20-45% SDG, Mixed with a second quantity of lignan extract, eg substantially pure lignan extract, ie having at least 90% SDG. US Patent 5,705,618 to Westcott et al. discloses a lignan extract with 90% SDG, the entire contents of which are incorporated by reference.

Table 11 discloses compositions comprising different concentrations of SDG (37.75%-88.8% SDG) by combining a first lignan extract with about 35% SDG with a second lignan extract with about 90% SDG Mix to produce.

Example 8

The molecular weight of the lignan product produced by Example 1 using centrifugation was determined by gel permeation (size exclusion) chromatography. Standard kits obtained from Phenomenex (Torrance, CA): poly(ethylene glycol) part number ALO-2774 and poly(ethylene oxide) part number ALO-2775 were used. Use Phenomenex PolySep-GFC-P 4000, 300x 7.8mm gel permeation chromatography column. A solvent mixture of 50% acetonitrile/50% water was used for elution at a flow rate of 0.350 mL/min. Standards were determined by evaporative light scattering detection. Samples were measured at 280 nm by UV-Visible detection. Class-VP software version 7.2.1 SP1 from Shimadzu Scientific Instruments with SEC (Size Exclusion Chromatography) package was used to calculate the molecular weight parameters of the samples. The chromatograms are shown in Figures 1A-1D.

Example 9

The components of the five lignan extracts produced according to this embodiment of the invention were analyzed and the results are provided in Table 12. Batches A-C were produced using the embodiment of Example 1 using coalescence/sedimentation, and batches D-E were produced using the embodiment of Example 1 using centrifugation.

All documents mentioned herein are hereby incorporated by reference.

Although the foregoing invention has been described in detail for purposes of clarity and understanding, those skilled in the art will, from a reading of this disclosure, appreciate that various changes in form and details can be made without departing from the scope of the invention and the appended claims. For example, although specific plant species, extracts, compositions, methods and uses thereof have been described, those skilled in the art will recognize that various chemical substitutions or changes to any exemplary embodiment are possible without departing from the invention and the scope of the appended claims. category. Furthermore, any components, compounds, compositions, or steps of the invention may be combined.

Claims (43)

1. compositions comprises:

The first lignan extract with the secoisolariciresinol diglucoside that is less than 50% content; With

Has the second lignan extract more than the secoisolariciresinol diglucoside of 50% content.

2. the compositions of claim 1, wherein compositions comprises the 50-600mg secoisolariciresinol diglucoside.

3. claim 1 or 2 compositions, wherein compositions is selected from food, beverage, capsule, tablet and capsule sheet.

4. each compositions among the claim 1-3 further comprises the chemical compound that is selected from sterol, isoflavone, sweeting agent, protein isolate, protein concentrate, non-nutritive sweetener, soluble fiber, insoluble fiber, Statins, vitamin E, proanthocyanidin, Saw Palmetto P.E, Pygeum extract, oleum lini, fish oil, vitamin B, vitamin C, tocopherol, plant sterol, polyphenol, catechin, anthocyanin, astaxanthin, glycosamine and combination in any thereof.

5. compositions comprises:

Lignan extract with 20-45% secoisolariciresinol diglucoside;

Wherein compositions comprises 50mg or lower cyanogenetic glucoside/100g compositions.

6. the compositions of claim 5, wherein compositions comprises the 300-600mg secoisolariciresinol diglucoside.

7. the container that contains each compositions among the claim 1-6, wherein container is with the labelling that instructs user compositions recommended dose.

8. the compositions of claim 1, wherein the first lignan extract is that the 30-40% secoisolariciresinol diglucoside and the second lignan extract are at least 90% secoisolariciresinol diglucoside.

9. the purposes of each compositions among the claim 1-6 or 8 is used to prepare the cholesterol reducing level, the treatment benign prostatauxe, treatment heart disease, prevention hypercholesterolemia arteriosclerosis, alleviate heart disease, reduce serum cholesterol, reduce ischemic lesions, strengthen nitric oxide expression in the endotheliocyte, suppress cyclooxygenase activity in the macrophage, alleviate diabetic symptom, prevention or the outbreak of delay diabetes, adjust serum level of glucose, bone density improving, reduce the bone density loss, the ameliorate osteoarthritis symptom, reduce the carcinoma of prostate risk, reduction shifts risk, postpone to shift, alleviate menstruation outbreak symptom, alleviate the symptom of showing effect menopause, the medicine of blood pressure lowering or minimizing alopecia or male pattern alopecia.

10. the lignan extract that comprises the 46-89% secoisolariciresinol diglucoside.

11. the purposes of the lignan extract of claim 10 is used to prepare the cholesterol reducing level, the treatment benign prostatauxe, treatment heart disease, prevention hypercholesterolemia arteriosclerosis, alleviate heart disease, reduce serum cholesterol, reduce ischemic lesions, strengthen nitric oxide expression in the endotheliocyte, suppress cyclooxygenase activity in the macrophage, alleviate diabetic symptom, prevention or the outbreak of delay diabetes, adjust serum level of glucose, bone density improving, reduce the bone density loss, the ameliorate osteoarthritis symptom, reduce the carcinoma of prostate risk, reduction shifts risk, postpone to shift, alleviate menstruation outbreak symptom, alleviate the symptom of showing effect menopause, blood pressure lowering, reduce the medicine of alopecia or male pattern alopecia.

12. cholesterol reducing level, the treatment benign prostatauxe, treatment heart disease, prevention hypercholesterolemia arteriosclerosis, alleviate heart disease, reduce serum cholesterol, reduce ischemic lesions, strengthen nitric oxide expression in the endotheliocyte, suppress cyclooxygenase activity in the macrophage, alleviate diabetic symptom, prevention or the outbreak of delay diabetes, adjust serum level of glucose, bone density improving, reduce the bone density loss, the ameliorate osteoarthritis symptom, reduce the carcinoma of prostate risk, reduction shifts risk, postpone to shift, alleviate menstruation outbreak symptom, alleviate the symptom of showing effect menopause, blood pressure lowering, reduce the method for alopecia or male pattern alopecia, comprise with claim 1-6 or 8 each compositions or the lignan extract of claim 10 deliver medicine to the patient.

13. cholesterol reducing level, the treatment benign prostatauxe, treatment heart disease, prevention hypercholesterolemia arteriosclerosis, alleviate heart disease, reduce serum cholesterol, reduce ischemic lesions, strengthen nitric oxide expression in the endotheliocyte, suppress cyclooxygenase activity in the macrophage, alleviate diabetic symptom, prevention or the outbreak of delay diabetes, adjust serum level of glucose, bone density improving, reduce the bone density loss, the ameliorate osteoarthritis symptom, reduce the carcinoma of prostate risk, reduction shifts risk, postpone to shift, alleviate menstruation outbreak symptom, alleviate the symptom of showing effect menopause, blood pressure lowering, reduce the method for alopecia or male pattern alopecia, the compositions that comprises the lignan complex that will have at least 100,000 molecular weight delivers medicine to the patient.

14. a method for the treatment of benign prostatauxe comprises that the lignan complex that will have at least 100,000 molecular weight delivers medicine to the patient.

15. a method that reduces blood in human body in clearing gallbladder sterin comprises that the lignan complex that will have at least 100,000 molecular weight delivers medicine to the patient.

16. the method for a blood pressure lowering comprises that the lignan complex that will have at least 100,000 molecular weight delivers medicine to the patient.

17. a method that reduces international prostate gland symptoms scoring comprises the lignan complex is delivered medicine to the patient.

18. each method of claim 12-17, wherein the patient is human.

19. each method of claim 12-18 is wherein carried out oral administration.

20. a method of producing the lignan extract comprises:

The vegetable material that will contain lignan contacts with extracting solvent, extracts solution thereby form;

Use is isolated the part that is rich in lignan based on the separation equipment of density from extract solution;

Use the aqueous solution flushing to be rich in the part of lignan; With

Drying is rich in the part of lignan.

21., comprise that further decant goes out part extraction solution from the part that is rich in lignan according to the method for claim 20.

22. according to each method of claim 20-21, wherein aqueous solution comprises the citrate aqueous buffer solution.

23., comprise that further removing a part from extract solution extracts solvent according to each method of claim 20-22.

24. the method according to claim 23 further comprises:

Wherein from extract solution, remove the extracting section solvent, comprise the vaporize draw solvent; With

Utilize the extraction solvent that evaporates again.

25., wherein extract solvent and comprise alcohol according to each method of claim 20-24.

26., further comprise and grind the part that is rich in lignan according to each method of claim 20-25.

27., comprise that further packing is rich in the part of lignan according to each method of claim 20-26.

28., comprise that further the part that will be rich in lignan places food, capsule, tablet, capsule sheet or beverage according to each method of claim 20-27.

29. according to each method of claim 20-28, the part that wherein is rich in lignan contains the secoisolariciresinol diglucoside of at least 35% weight.

30. according to each method of claim 20-29, the part that wherein is rich in lignan comprises that 50mg or lower cyanogenetic glucoside/100g are rich in the part of lignan.

31. according to each method of claim 20-30, wherein the separation equipment based on density is selected from hydrocyclone, subsider, centrifuge and filter.

32. according to each method of claim 20-31, the wherein dry part that is rich in lignan is carried out in the exsiccator that is selected from vacuum band exsiccator, microwave dryer, radio-frequency seasoning device and freeze dryer.

33., further comprise the vegetable material that is selected from screening and contains lignan, vegetable material that sucking-off contains lignan or the behavior of its combination according to each method of claim 20-32.

34. the method for claim 33 wherein uses two wire screens to screen the vegetable material that contains lignan.

35. the method for claim 34, wherein two wire screens comprise one 500 microns sieve and one 2500 microns sieve.

36., further comprise the part that to be rich in lignan and the compound that is selected from sterol, isoflavone, sweeting agent, protein isolate or concentrate, non-nutritive sweetener, solvable or insoluble fiber, Statins, vitamin E, proanthocyanidin, Saw Palmetto P.E, Pygeum extract, oleum lini, fish oil, vitamin B, vitamin C, tocopherol, plant sterol, polyphenol, catechin, anthocyanin, astaxanthin, glycosamine and combination in any thereof according to each method of claim 20-35.

37. product by each method production of claim 20-36.

38. the purposes of the product of claim 37 is used for the cholesterol reducing level, the treatment benign prostatauxe, treatment heart disease, prevention hypercholesterolemia arteriosclerosis, alleviate heart disease, reduce serum cholesterol, reduce ischemic lesions, strengthen nitric oxide expression in the endotheliocyte, suppress cyclooxygenase activity in the macrophage, alleviate diabetic symptom, prevention or the outbreak of delay diabetes, adjust serum level of glucose, bone density improving, reduce the bone density loss, the ameliorate osteoarthritis symptom, reduce the carcinoma of prostate risk, reduction shifts risk, postpone to shift, alleviate menstruation outbreak symptom, alleviate the symptom of showing effect menopause, blood pressure lowering or minimizing alopecia or male pattern alopecia.

39. the purposes of the product of claim 38 is used to prepare the cholesterol reducing level, the treatment benign prostatauxe, treatment heart disease, prevention hypercholesterolemia arteriosclerosis, alleviate heart disease, reduce serum cholesterol, reduce ischemic lesions, strengthen nitric oxide expression in the endotheliocyte, suppress cyclooxygenase activity in the macrophage, alleviate diabetic symptom, prevention or the outbreak of delay diabetes, adjust serum level of glucose, bone density improving, reduce the bone density loss, the ameliorate osteoarthritis symptom, reduce the carcinoma of prostate risk, reduction shifts risk, postpone to shift, alleviate menstruation outbreak symptom, alleviate the symptom of showing effect menopause, the medicine of blood pressure lowering or minimizing alopecia or male pattern alopecia.

40. a method of removing cyanogenetic glucoside from Caulis et Folium Lini extract comprises:

With the chemical compound flushing Caulis et Folium Lini extract that is selected from citric acid, ethylenediamine tetraacetate, acetic acid, Fumaric acid, lactic acid and its combination in any.

41. the method for claim 40, wherein this chemical compound has the pH of 3.0-6.0.

42. one kind increases enterolactone, enterodiol or its method for compositions in patient's blood plasma, comprising:

With the claim 1-6 of effective dose or 8 each compositions or the product of claim 37 deliver medicine to the patient.

43. one kind increases enterolactone, enterodiol or its method for compositions in patient's blood plasma, comprising:

With 300mg/ days to 600mg/ days claim 1-6 or 8 each compositions or the product of claim 37 deliver medicine to the patient.

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