Stanilova composition and its application

 

(57) Abstract:

The invention relates to medicine. The invention is characterized by the fact that stanilova composition containing in addition to the main component of sitostanol at least 10% of campestanol, when introduced into edible products effectively lowers the level of cholesterol in blood serum. In esterified form, the composition is particularly useful in edible fats and oils and fat-containing foods. 3 s and 5 C.p. f-crystals, 4 PL.

The present invention relates to the composition of plant stanilov containing sitostanol and specially designed for use as a drug, lowering the level of cholesterol in blood serum. The invention relates also to the corresponding esterified form of such a composition, which has advantages when used in edible oils and fats, and fat-containing foods. The invention relates also to a method for producing food products with the use of this composition.

Art

Plant sterols are essential components of all plants. Their function in plants is similar to the functions of cholesterol in mlekopitayushchie. The chemical structure of these plant sterols are very similar to the structure of cholesterol with differences in the side chain of the skeleton of the molecule. For example, compared with the cholesterol side chain of sitosterol and campesterol additionally contain, respectively, ethyl and methyl groups.

Since the 1950's it was known that plant sterols effectively lowers cholesterol levels in the serum. Even with the introduction of relatively small doses (several grams per day), they effectively reduce the ability to absorb both biliary and dietary cholesterol, thus reducing the levels of total and LDL-cholesterol in serum (12, 28, see also 27, 32). The mechanism by which limits the absorption of cholesterol, so far the details are unknown, but assumed that plant sterols displace cholesterol from the micellar phase and thus prevent its absorption. Almost all earlier studies a major study plant Sterol was sitosterol or hydrogenomonas form sitostanol. However staronova composition of the tested drugs was not always well predocumentation, and applied in most studies Stereolove pros as a safe method of reducing cholesterol in the serum, because they are natural components of vegetable fats and oils. In addition, their absorption from the intestine of a healthy subject is limited, and limited the absorbed amount excredits from the body in bile. The degree of absorption of plant sterols varies from subject to subject and varies depending on the form of plant Sterol, but for healthy people from the digestive tract is usually absorbed less than 5% of plant sterols (27). However, it has been shown (20) that dietary campesterol is absorbed in an amount up to 10%.

In some rare diseases, such as sitosterolemia, plant sterols are absorbed very effectively, also violated their excretion from the body by the biliary route. Serum levels of sitosterol, campesterol, and their saturated form, i.e. sitostanol and campestanol dramatically increase. Elevated levels of saturated stanilov are most likely due to their more efficient endogenous synthesis, and not due to more efficient absorption (10, 27). In the absence of treatment sitosterolemia already at a young age leads to xanthomatosis and coronary heart disease. For people with this disease introduction unsaturated plant Sterol is factam.

Lees and Lees (25) was influenced by three different sitosterolemia drugs on the concentration of lipids and lipoproteins in the plasma. One of these drugs was Cytellin, industrial product (Eli Lilly Co., USA) containing 60-65% of sitosterol and 35-40% other sterols, mainly campesterol. The average dose of 18 g/day divided into three doses, led to a decrease in the total cholesterol in plasma on average by 10.5% and lower LDL cholesterol by 15%. However, if usually in the plasma are detected only traces of plant sterols, including campesterol (10, 33), the subjects who participated in the tests conducted Lees and Lees, concentration campesterol in plasma ranged from 4 to 21 mg/DL. When discussing the results of the authors very strongly argued that because the atherogenicity of campesterol unknown, application sitosterolemia drug with a relatively high content of campesterol, such as used in their study drug Cytellin, cannot be recommended.

Further Lees et al. (26) studied the effectiveness of plant sterols from oils derived from soybeans, and tall oil for lowering cholesterol levels in the blood. For each plant Sterol they used two rerole, and average daily dose of 18 g of sterols per day (doses 9-24 g) was given in three equal doses. This study used the drug Sterol from tall oil, containing only about 5% of campesterol. For both stalowych drugs from tall oil (powder and suspension) was tested daily dose of 3 g were also tested dose of 6 g of the suspension of Sterol from tall oil.

Soy Sterol in both physical form and sterols from tall oil in the form of powder lowered the cholesterol content in the plasma, on average, 12% (26). However, already shown previously relatively high ability campesterol absorption was also observed in this study. Five of the tested patients, the levels of campesterol in plasma ranged from 5 to 21 mg/DL (average 16 mg/DL). Again, even for the case proved a significant effect of lowering cholesterol with soy Sterol authors do not recommend its use as a reagent, reducing cholesterol. On the contrary, they recommend that pharmaceutical preparations of plant sterols contained a minimum of campesterol and maximum sitosterol. Based on the above two studies, one can conclude that the use of CTE is sisanie plant sterols, such as sitostanol and campestanol, are present in most vegetable oils only in trace amounts. However sterols from tall oil contain 10-15% of sitostanol saturated form of sitosterol. Sitostanol can also be obtained by hydrogenation of a double bond in the sitosterol. In subsequent studies, conducted in experimental animals and in humans, it has been shown that, as a reagent, lowering cholesterol, sitosterol more effective than sitosterol(8, 16, 17, 18, 19, 36).

An additional advantage of sitostanol is that it is not actually absorbed. Several studies (for example, 9, 16, 17, 21) showed that sitostanol almost not able to be absorbed, while a small amount (< 5%) unsaturated form of sitosterol (33) can be absorbed. Similarly in the in vitro study Amstrong and Carey (6) also showed that cholestanol, the saturated form of cholesterol, was more hydrophobic and less able to absorb than cholesterol.

Upon receipt of sitostanol by hydrogenation of the most common sources of plant sterols from campesterol forms as well as other saturated plant Sterol, namely campestanol. Until recent times the sustained fashion a bit. Based on the above data, claiming that saturated sterols have less ability to absorb than their unsaturated form, it would be in order hypothesis to conclude that campestanol can be actually not able to absorb.

To study the ability of different plant Sterol absorption Heinemann et al. (20) using the technique of perfusion of the intestine compared the absorption of cholesterol in the intestines of humans with similar effect to campesterol, sitosterol, stigmasterol, and for low concentrations of sitostanol and campestanol. The results showed that the degree of absorption of a variety of plant Sterol changed in the group of various plant sterols, an average of 4.2% for sitosterol, 4.8% of stigmasterol and 9.6% for campesterol and 12.5% for campestanol. For ten experimental males were identified large variations in the efficiency of absorption.

Thus, according to Heinemann et al. (20) have shown that campestanol absorbed more efficiently than its unsaturated form - campesterol. This contradicts the assumption, based on the above studies, which showed that saturated sterols (sitosterol, Ho is on this remains outstanding, although Heinemann et al. (20) suggested that the reason for this contradictory result may be that, firstly, the study Amstrong and Carey (6) was carried out in vitro and, secondly, theory of hydrophobicity as the main factor in micellar binding and/or uptake cannot be applied in vivo. However, this assumption does not explain the fact that several studies showed for sitostanol lower absorption capacity compared with this option for sitosterol, were carried out in vivo. Thus, the results of Heinemann et al. (20), contradicting the previous results, the authors remain unexplained.

Sugano et al. (34) studied hypocholesterolemic activity corn sterols (composition: 31% campesterol, 4% stigmasterol and 65% of sitosterol) and corn stanilov (composition: 31% campestanol and 69% of sitostanol) obtained by hydrogenation of a mixture of sterols from corn oil. Were conducted two experiments on rats. When taking cholesterol inside (1% of diet) as a Sterol, and stanol showed hypocholesterolemic effects at the level of 0.5-1% of the diet. In the first experiment, significant differences in hypocholesterolemic the effect of Phi is octanol compared with phytosterols showed significantly greater ability to decrease the concentration of cholesterol in plasma (statistically significant at p < 0,02). Moreover, rats fed at 1.0% Stanovoy diet had significantly lower cholesterol levels in plasma (p < 0,02) than animals fed on a diet free of cholesterol. This effect was not observed in rats fed at 1.0% stealaway diet.

Sugano et al. (34) did not investigate the difference hypocholesterolemic effect between mixtures stanilov with a high content of sitostanol and low content of campestanol (on the basis of Sterol from tall oil) and mixtures of Stanlow with significantly higher levels of campestanol (on the basis of Sterol from vegetable oil). They compared hypocholesterolemic effect of the mixture of unsaturated sterols and appropriate mixture of saturated stanilov. Later studies conducted by this research group focused on the effect of lowering cholesterol with sitostanol as itself and in comparison with data on sitosterol(21, 22, 23, 35). In fact, in a later publication (23) they refer to the above research phytostanol (34), only mentioning hypocholesterolemic effect sitostanol in comparison with-sitosterol without discussion of any hypocholesterolemic effect saturated with criminalise the above mixture of sterols with a typical composition hydrogenating sterols from tall oil with a high content of sitostanol (> 90%).

Miettinen and Vanhanen (30) showed that lowering the level of cholesterol in the blood serum of sitostanol in the form of ether fatty acids are more effective than free sitostanol. More recent studies have also shown that the use of esters of sitosterol as a component of the daily diet is an effective way of lowering the concentrations of total cholesterol and LDL-cholesterol in serum(13, 14, 15, 31, 37, 38). The advantage of using esters of stanol instead of free stanol also lies in the fact that the esters of stanol fat-soluble and therefore can be readily implemented in a wide range of foods without changing the taste, smell or physical conduct of the final product. The method of preparation of esters formed by sitosterol and fatty acids, and the use of fat-soluble esters of Stanlow in food have been described in U.S. patent N 5502045 (2) included in this description by reference thereto.

Straub (3) suggested the use of saturated stanilov (sitostanol, clionasterol, 22,23-dehydrobrominated, campestanol and their mixtures) in the preparation method of the composition of food additives, in which stanely mixed with edible solvent reagent, the effect is sevie additives are designed to reduce the absorption of cholesterol from food and drinks, contain cholesterol, such as meat, eggs and dairy products. However, in this patent is not provided data showing whether any clinical effects or absorption of dietary sterols.

Eugster et al. (1) describe the application of small amounts of sterols, fatty acid esters and glucosides for the treatment of tumors. Methods of cooking proposed by Eugster et al., include harmful chemical reagents, such as N,N'-carbonyldiimidazole, thionyl chloride and the solvent, such as tetrahydrofuran, benzene, chloroform or dimethylformamide. Eugster et al. comment on the possible use of these drugs as diet food and nutritional supplements, but does not give any data about hypocholesterolemic effects and not formulate any suggestions for this application. From the description of Eugster et al. it is difficult to get a clear picture of how to purify the end product from obtaining sufficiently pure ester Sterol in quantities large enough for use as a food component. The only proposed treatment processes are thin-layer chromatography and high performance liquid chromatography. Anyway the>/P>U.S. patent N 3751569 (4) describes the addition of esters formed with plant sterols and fatty acids, to cooking oil to reduce cholesterol levels in the serum of a person. For use in the esterification of free sterols patent proposes a method that is not in all respects comply with the requirement imposed on the preparation of the food product series. According to the patent etherification occurs between the free Sterol and the anhydride of the fatty acid in the presence of perchloro acid acting as a catalyst. The applied catalyst and reagent may not be allowed to participate in food processes. In addition, the patent refers to esters formed fatty acids and natural plant sterols. The method proposed in German patent N 2248921 (5) for the esterification of sterols present in oils and fats by chemical esterification meets the criteria attached to food processes. In this patent the free Sterol and an excess of fatty acid esters is added to a mixture of oil or fat, then all the fat mixture interesterification using a conventional known technology interesterification. In the fat blend of the actual adnych sterols in vegetable and animal oils from possible changes during the process.

Earlier data show that campesterol, one of the major plant sterols, is absorbed relatively efficiently. Therefore, the application had been recommended to mix only plant sterols with a minimum content of campesterol. In practice, this leads to the use of mixtures of sterols, such as sterols from tall oil with a high content of sitosterol.

Most of the works on stazolam concerned only sitostanol. The study Heinemann et al. (20), showing that campestanol, the saturated form of campesterol, is absorbed more easily than campesterol or sitosterol (respectively 12,5, 9.6 4.2%), and resulted in a "consensus" about the fact that a mixture of saturated sterols with "elevated" levels of campestanol due to absorption campestanol unsafe. The obvious proof of this is the fact that all clinical studies regarding the application of stanilov (sitostanol) was based on mixtures of sterols with a high level of sitostanol and low campestanol.

Many studies (e.g., 8, 17, 18, 19, 23, 36) established that higher efficiency sitostanol saturated form of sitosterol, compared to the corresponding unsaturated sitosterolaemia, making use of saturated sterols safe means of lowering cholesterol across the population. Of all unsaturated sterols in particular campesterol absorbed in quantities in a high degree sufficient to formulate recommendations against the use of mixtures of sterols with elevated levels of campesterol (for example, mixtures of sterols on the basis of vegetable oils) (25, 26).

According to said there was a strong prejudice against the use of campestanol in any significant quantities as a drug that is added to food, and this seriously limited the range of raw materials that contain phytosterols, compounds with a relatively small number of campesterol and vivid form, of campestanol.

The invention

The present invention relates to compositions of plant stanilov, as a main component containing sitostanol, but a significant number of campestanol, in free form or esterified to fatty acid esters, and intended to reduce the level of cholesterol in blood serum. The invention relates also to the use of a mixture of phytosterols, vklyucheniya this composition.

In addition, the invention relates to food products and to the method of obtaining these drugs on the basis of stanulovic compositions, as a main component containing sitostanol, but a significant number of campestanol, or their fatty acid esters in edible products in the form of a dietary component that is intended to reduce the level of cholesterol in blood serum.

The problem solved by the present invention, is to expand the number of vegetable raw materials suitable for the preparation of drugs for edible products, particularly edible oils and fats and fat-containing foods, and intended to regulate the level of cholesterol in blood serum. The invention can be used as raw materials for these purposes and vegetable oils and fats containing in addition to sitosterol also a significant number of campesterol.

Suitable raw materials for use in the preparation of compositions of the present invention are, for example, corn, soybeans and canola, but can also be used and other plant-derived substances with phytosterolemia composition with a high content of the Orme, can be put into food substances such as cooking oil, margarine, butter, mayonnaise, salad garnishes added to the dough fats, cheeses (including unripe and ripened cheeses) and other fat-containing foods.

The composition according to the present invention can also be used by itself.

According to the present invention, the composition of plant stanilov, in addition to its primary component to sitostanol, also contains a significant amount, at least 10%, campestanol.

The composition preferably contains campestanol (from 20 to 40%, most preferably from 25 to 35%, for example about 30%) or fatty acid ester in the case where the composition is to give her liquid was subjected to esterification.

In the present description, all percentages are given as percent by weight except where otherwise noted, numbers in brackets refer to the publications placed in the attached reference list.

The data obtained, unusual and contrary to common belief, show that hydrogenomonas mixture of Stanlow containing as a main component sitostanol, but significant Klah and low campestanol, indicating that campestanol at least as efficient in lowering the absorption of cholesterol, as sitostanol. Moreover, the data analysis of Sterol serum clearly show that campestanol remains virtually unabsorbed, while its content in the serum approximately 40% less than the content of sitostanol. Thus, a mixture of Stanlow containing as a main component sitostanol, as well as appreciable quantities of campestanol, should be considered at least as safe as conventional mixture of Stanlow on the basis of tall oil sterols. These data are in sharp contradiction with the common opinion concerning the efficacy and safety of mixtures of Stanlow with elevated amounts of campestanol (see 20, 27, 34).

In U.S. patent N 5502045 (2) shows that the esters formed by sitosterol and fatty acids, more effective in lowering cholesterol levels in the blood than the free sitostanol. Subsequent studies clearly confirmed the effect of lowering cholesterol margarine containing fat-soluble esters formed by sitosterol and fatty acids (see, e.g., 31).

The use of esters formed statereason food because only the esters formed by stanolone and fatty acids, soluble in edible oils and fats in quantities large enough to achieve levels that are effective in lowering the absorption of both dietary and biliary cholesterol from the digestive tract.

Solubility stanulovic esters in edible oils and fats is 35-40%, while the solubility of free sterols in these objects is limited only by the maximum of 2% by weight at a temperature of 21oC (24). A higher number could be introduced by using different surfactants and reagents solvent or dispersant type, but the use of such substances does not provide solubility in fats. The use of the above drugs is usually limited or even prohibited by law. In addition, free sterols at the 1% level will affect the physical behavior of the product. The situation here is different from the application of the esters formed by stanolone and fatty acids, because the physical properties of the fat blends can be easily modified by changing the composition of fatty acids mixture.

Obviously, the esters formed by stanolone and fatty acids, can be legcom the invention. In U.S. patent N 5502045 (2) are other examples of possible applications. For specialists in this area, however, it is obvious that the esters formed by stanolone and fatty acids, can be added to a wide variety of food, especially fat-containing food.

It was suggested many ways of obtaining fatty acid esters of sterols. The disadvantages of these methods is that almost all of them use reagents that cannot be allowed in a product intended for use as micropitting component in food. Commonplace is the use of such toxic reagents like thionyl chloride or anhydrous derivatives of fatty acids.

The preferred method of obtaining stanulovic of fatty acid esters on the basis of sterols described in U.S. patent N 5502045 (2 included in this description by reference thereto). This method is based on the process of interesterification, widely used in the manufacture of edible fats and oils. From the previous methods, this process of esterification is different the advantage that it does not use any drugs in addition to free stanol, ether fatty colornotes method is the fact, one of the reagents, namely fatty acid ester, used in excess and functions as a solvent, dissolving stanol when used conditions (vacuum 5-15 mm Hg). The reaction produces a mixture of fatty acid esters and ethers formed by stanolone and fatty acids. The ether formed by stenolol and fatty acid, can be easily concentrated in almost pure consistency by vacuum distillation to remove the excess fatty acid esters. In the alternative, the mixture itself can be added to the final fat mixture before carrying out stage deodorization.

Small amounts of stanely found in nature, for example, in wheat, rye, corn, triticale and, thus, in small quantities they can be found (11, 14) in the daily diet. Stanely can be easily obtained by the hydrogenation of mixtures of natural sterols. In the beginning of 1996 in the commercial sale had only a mixture of tall oil sterols with high purity (Sterol levels > 98%), designed by themselves for food applications. Plant sterols with significant quantities of campesterol, such as a mixture of sterols on the basis of vegetable oils can be obtained, for example, as a by-product p is owani in stanely using well-known technologies hydrogenation such as technology based on the application of Pd/C catalyst in organic solvents (publication 7, introduced in this description by reference to it). For specialists in this area it is obvious that a wide variety of Pd catalysts and solvents can be used to implement the hydrogenation, which in optimal conditions, leaves non-convertible only small amounts of unsaturated sterols, whereas the formation of typical dihydroxypropane products stanilov and sterols remains at a low level (< 1.5 percent).

In the framework of the present invention is compared hypocholesterolemic effect of a mixture of Stanlow containing a high level of sitostanol, usually considered experts in this area the safest and the most efficient plant Sterol from the viewpoint of reducing absorption of cholesterol and, through this, the level of cholesterol in the serum, with the same effect of a mixture of Stanlow, containing a significant number of campestanol. In the present description for the first time reported hypocholesterolemia impacts Stanlow on the basis of vegetable oils on people. In the present description for the first time shown is e as effective as a mixture of Stanlow with high levels of sitostanol. Moreover, the results of this study clearly indicate that campestanol, contrary to the data reported by Heinemann et al. (20), is not actually absorbed.

Clinical studies

To study hypocholesterolemia impacts Margarines with esters of Stanlow on the basis of vegetable and tall oil was taken 5-week crossover study double-blind method with a 2-week period of the washing cycle. Test study design was as follows.

Test study design exposure.

Rooms 1-6 indicated blood samples collected at home food (1, 2), after the first period of exposure (3, 4) and after the second period of exposure (5, 6). OC - margarine with ether stanol based on vegetable oil, TC - margarine with ether stanol on the basis of tall oil.

< / BR>
Twenty-four volunteer, non-performing healthy women with moderately elevated cholesterol levels (average 6,12 0.16 mmol/l) took approximately 25 g / day (250 gram tube/10 days) of passing the test margarine as part of the daily diet in a random order. The serum lipids (total Masnum nutrition and at the end of each test period. Blood samples were taken twice, after one week at home meals and at the end of each test period margarine. The obtained values of lipids in the serum are presented in table 1.

For both the tested Margarines were found positive changes in serum lipids. The lower values for LDL-cholesterol and increasing values for HDL-cholesterol were statistically significant (p < 0.05 or less). In addition, Sterol ester-based vegetable oil in statistical processing of results was also shown to decrease total cholesterol. The resulting decrease in the total cholesterol and LDL-cholesterol was higher for margarine with ether stanol on the basis of vegetable oils compared to margarine with ether stanol on the basis of tall oil. For levels of triglycerides changes are not received. The results obtained for serum lipids, show that margarine with ether stanol on the basis of vegetable oils containing in its Stanovoy faction significant number of campestanol, can be even more effective than margarine with ether stanol on the basis of tall oil. Margarine with ether stanol on the basis of tall oil in more early cross circuit of the present study can be concluded, that stanely based on vegetable oils show at least the same hypocholesterolemia impact that stanely on the basis of tall oil.

The concentration of sterols in serum were quantified by gas-liquid chromatography according to previously published method (publication 29 included in the present description by reference to it). Calculated average values of the results of two measurements of lipids in the serum from blood samples taken at each period. In tables 2 and 3 presents data on the average concentrations of plant sterols in serum at home diet and after each test period, and averaged changes observed at these concentrations.

Both have been tested margarine was significantly lowered the levels of campesterol and sitosterol in the serum. It is known that in humans, the concentration of campesterol in serum correlates with the absorption of cholesterol in the intestine (29, 39). Thus, the lower the value of cholesterol, the lower the absorbed percentage of cholesterol in the intestine.

A marked drop in the levels of campesterol in serum (25 - 28%) during periods of studies is also could not find any differences in the concentration of sitostanol in the serum, while the average concentration of campestanol in serum after a period of exposure to ether stanol on the basis of vegetable oils was significantly higher than at home diet and after a period of exposure to ether stanol on the basis of tall oil. However, the absolute concentration of campestanol was only about 63% of the concentration of sitostanol, which is usually considered practically insoluble substance. Such a low concentration of campestanol in serum clearly indicates the very limited absorption campestanol that is in contradiction with the results presented Heinemann et al. (20). Thus, because of the mix of Stanlow containing high levels of sitostanol, are considered safe for humans by ingestion, a mixture of Stanlow containing substantial amounts of campestanol, should be considered as safe on the basis of the fact that campestanol like sitostanol actually has no ability to absorb.

The following working examples describe the preparation of compositions of esters stanilov according sobrosa the possibility of carrying out the invention

Example 1. Hydrogenation of mixtures of sterols.

Commercially available mixture of sterols obtained from distillate vegetable oil (ingredients: brassicasterol 2.7%, campesterol 26,7%, stigmasterol 18,4%, sitosterol 49.1% and sitostanol 2,9%) were hydrogenosomal in the pilot reactor (25 I). The reactor was placed 26 g fiber Pd-catalyst (Smop-20; Pd content 10% by weight, Smoptech, Finland), 26 g of distilled water to activate the catalyst and 11.7 kg propanol. The reactor was filled with nitrogen and held the activation of the catalyst under gaseous hydrogen at a pressure of 1 bar and a temperature of 65oC within 30 minutes After activation, the mixture was cooled to 40oC, and then added 1.3 kg of a mixture of sterols.

A mixture of propanol and sterols heated from the temperature of the nitrogen atmosphere to 65oC, after which the nitrogen was replaced by hydrogen. After a full fill of hydrogen, was the reaction of hydrogenation under hydrogen pressure of 1 kgf/cm2. The normal conversion time is approximately 120 minutes the Transformation can be easily controlled by the selection of doses that are analyzed by HPLC.

The hydrogen pressure dropped and the reactor was filled with nitrogen. Fibrous catalyst otherthrow the>, after which the crystals Stanlow was filtered with vacuum and the residue was washed in 0.5 kg of cold propanol. The mixture of Stanlow dried at 60oC in a vacuum drying Cabinet. The yield was 75%, and the composition of the mixture of Stanlow according to capillary GC analysis was as follows: campesterol of 0.2%, campestanol of 28.9%, stigmasterol 0.1%, sitosterol 0.2%, sitostanol 70,1%. It should be noted that brassicasterol hydrogenized 24-methylgalactose, the epimer of campestanol, however, because the operations of conventional capillary gas chromatography it is manifested in the form of the same peak, which according to the chirality cannot be split, it is usually calculated as campestanol. If we start from the initial mixture of sterols, 24-methylgalactose would have amounted to 2.7%.

Example 2. Preparation of esters formed by stanolone and essential fatty acids.

The mixture of esters formed by stanolone and fatty acids, prepared in the pilot plant. 6 kg of Stanlow resulting from the combination of several parties, processed as described in example 1, the procedure of hydrogenation was dried over night at 60oC and atrificial with 8.6 kg of a mixture of methylated esters formed by the tin: campesterol of 0.4%, campestanol (+ 24--methylgalactose) 29,7%, stigmasterol 0.1%, sitosterol 0.4% and sitostanol to 68.0%. The content of Stanlow in the mixture was 98.2%. The etherification was carried out as follows.

A mixture of Stanlow and methylated ester formed by the lower erucic fatty acid based on rapeseed oil, kept in heated condition in the reaction vessel at 90-120oC under vacuum 5-15 mm Hg. After drying for 1 hour was added 21 g of ateleta Na and the reaction continued for about 2 hours. The addition of water (30% by weight) at 90oC was destroyed catalyst. After phase separation the aqueous phase was removed and conducted a second washing. After separation of the aqueous phase, the oily phase was dried with vacuum at 95oC with a stirring speed of 200 rpm, a Mixture of esters formed by stanolone and fatty acids, slightly bleached for 20 minutes at 30 mm Hg and a temperature of 110oC with the aid of bleaching earth (1,0%) (Tonsil Optimum FF, firm Sudchemie, Germany) under stirring at 200 rpm Whitening land was filtered, and the resulting mixture was methylated esters of fatty acids and esters formed by stanolone and fatty acids, may be added by itself to the fat smetana, the mixture can be deodorized with getting no taste of the mixture of esters formed by stanolone and fatty acids, which in itself can be added to various processes of food production.

The conversion process of esterification is usually > 99%, according to fast HPLC and the yield is about 95%.

Example 3. The production of Margarines for clinical trials.

80% of Margarines with esters formed by stanolone on the basis of tall oil and fatty acids, and esters formed by stanolone on the basis of vegetable oils and fatty acids were produced in the pilot plant (Perfector) Gerstenberg &Agger 3 x 57. Esters formed by stanolone on the basis of tall oil and fatty acids were derived from conventional margarine Benecolcompany Raision Margarini, Finland. Applied normal TRANS-mixture of fats, free fatty acids (composition: 30% dehydrogenating interesterification vegetable fat and 70% of liquid oil LEAR), to which was added a mixture of esters formed by stanolone and fatty acids. It was planned to have the contents of Stanlow in the final product 12 g/100 g of the product that would BR>
The mixture of fats that include esters formed by stanolone and fatty acids - 80%

Water - 19%

Sol - 0,5%

Emulsifier, Dimodan BP

The Na bicarbonate and citric acid as reagents,

regulating pH

-Carotene as a reactant-dye

Flavour

Received Margarines were Packed in polypropene tubes, weighing 250 g, protected with aluminum foil. Taste and density of the food was the equivalent parameters of industrial Margarines.

The content of Stanlow was 12.7 g/100 g margarine with stenolol based taly oil and 12.6 g/100 g margarine with stenolol based on vegetable oil. Staronova the composition of the two products are shown in table 4.

Sources of information

1. Eugster, C. , C. Eugster, Haldemann W., G. Rivara Sterols, fatty acid esters and glucosides; methods for their preparation; spontaneously dispergirujutsja reagents containing these compounds, and their use for the treatment of tumors. 1993. U.S. patent N 5270041.

2. Miettinen, T. A. , Vanhanen H., Wester I. application of the esters formed by stanolone and fatty acids, to lower the level of cholesterol in blood serum. 1996. U.S. patent N 5502045.

3. Straub C. D. Stanol the HT USA N 5244887.

4. Sterile cooking and salad oil that has hypocholesterolemic properties. 1973. U.S. patent N 3751569.

5. Baltes J., Merkle R. a Method of producing mixtures of vegetable and animal oils, such as fats and fatty acid esters. German Patent N 2248921.

6. Amstrong, M. J., Carey, M. C. Thermodynamic and molecular factors solubilities of sterols in the micelles of bile salts. J. Lipid Res. 1987; 28: 1144-1155.

7. Augustine R. L, Reardon Jr. E. J. 1969. Palladium catalyzing the hydrogenation of cholesterol. Org. Prep. and Proced. 1969; 1: 107-109.

8. Becker M., Staab d, von Bergmann K. Treatment of acute familial hypercholesterolemia in childhood with sitosterol and sitostanol. J. pediatr. 1993; 122: 292-296.

9. Czubayko F., Beumers Century, Lammsfuss S., Lutjohann d, von Bergmann K. Simplified microspool quantitative analysis of fecal excretion of neutral and acidic sterols for outpatient studies of people. J. Lipid Res. 1991; 32: 1861-1867.

10. Dayaj B., Tnt, G. S., P.le A. K., J. Speck, Khachadurian A. K., Schefer, S., Salen G. Identification of 5-Stanlow in patients with sitosterolemia and xanthomatosis: stereochemistry protonation steroid organoboron. Steroids 1982; 40:233-243.

11. Dutta, P. C., L. A. Appelqvist Saturated sterols (stanely) in dehydrogenating and hydrogenating edible vegetable oils and lips on the absorption of cholesterol in man. In: Greten H. (Ed.). Metabolism of lipoproteins. Springer-Verlag, Berlin, Heidelberg, New York, 1976: 112-118.

13. Gylling H. , Miettinen, T. A. Cholesterol serum, reduce dietary sitostanol, correlates with reduced absorption and synthesis of cholesterol and reducing the transport of LDL apoprotein b in people with type II diabetes. In: Gotto Jr. A. M., Mancini, M., Richter, W. O., Schwandt P. (Eds.). Acute treatment of dyslipoproteinemia in the prevention of coronary heart disease. 4th Int. Symp. Munich 1992, Karger, Basel, 1993:57-59.

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1. The composition of the esters formed vegetable stanolone and fatty acids intended for use as drug lowering cholesterol levels in blood serum, comprising as a main component an ester formed by sitostanol and fatty acid, and containing at least 10% ether formed by campestanol and fatty acid.

2. Composers who CLASS="ptx2">

3. The composition according to p. 2, characterized in that it contains from 25 to 35% ether, formed by campestanol and fatty acid.

4. The composition according to p. 3, characterized in that it contains about 30% ether formed by campestanol and fatty acid.

5. Composition according to any one of paragraphs.1 to 4, characterized in that it contains from 50 to 80% ether formed by sitostanol and fatty acid.

6. The method of obtaining food preparation by adding to food, such as fat-containing products, component of a diet for lowering cholesterol in blood serum, characterized in that, as specified component using the composition according to any one of paragraphs.1 - 5.

7. A food product that contains effective in lowering cholesterol in blood serum composition of esters formed vegetable stanolone and fatty acids, which contains the ester formed by sitostanol and fatty acid, and optionally contains ether formed by campestanol and fatty acid, the weight ratio of ester campestanol to air of sitostanol from 1:9 to 4:6.

8. Food drug on p. 7, characterized in that said weight ratio is from 2:8 to 3.5 to 6.5.

 

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