MXPA99001350A - Stanol composition and the use thereof - Google Patents

Abstract

A stanol composition containing in addition to sitostanol as the main component, also a substantial amount of at least 10%campestanol has been found to effectively lower serum cholesterol levels when incorporated in edible commodities. Upon esterification the composition is especially useful in edible fats and oils and in fat-containing foods.

Description

CO MPOSITION OF ESTANOL AND THE USE OF THE SAME Field of the invention The present invention relates to a composition containing sitoestanol from plant stanols especially for use as a substance that lowers the level of cholesterol in serum. The invention also relates to the corresponding esterified form of said composition which can be advantageously used in edible oils and fats and in foods containing fats. Background to the invention Plant sterols are essential components of all plants. Its functions in plants resemble the functions of cholesterol in mammals. The sterols of plants abundant in the flora are β-sitosterol, campesterol and stigmaesterol. The chemical structure of these plant sterols is very similar to that of cholesterol, the differences are presented in the side chain of the structure of the base of the molecule. For example, compared to cholesterol, the side chain of sitosterol contains an additional ethyl group and the side chain of campesterol has an additional methyl group. Since 1950, it has been known that plant sterols effectively reduce serum cholesterol levels. Even when administered in relatively small doses (a few grams per day), they effectively reduce the absorption capacity of both bile and dietary cholesterol and, therefore, lower total serum and LDL-cholesterol levels (12, 28, see also 27, 32). The mechanism by which restriction of cholesterol absorption is present is not yet known in detail, but it is assumed that plant sterols displace cholesterol from the micellar phase and therefore prevent its absorption. In practically all early studies, sitosterol or its sitostanol in hydrogenated form have been the plant sterol of main interest. However, the sterol composition of the tested preparations has not always been well documented and the sterol preparations used in most studies have also contained different amounts of other sterols. Plant sterols have been considered as a safe way to lower serum cholesterol levels, because they are natural components of vegetable fats and oils. Additionally, their absorption from the intestine of healthy subjects is limited, and the amounts absorbed are excreted from the body in the bile. The absorption regime of plant sterols varies between individuals and between different plant sterols, but for healthy humans usually less than 5% plant sterols are absorbed from the digestive tract (27). However, up to 10% of dietary campesterol has been shown to be absorbed (20). In a few rare diseases, such plant sterols of sitoesterolemia are exceptionally absorbed efficiently and also the elimination of the body via the biliary route is interrupted. The serum levels of sitosterol, campesterol and also its saturated forms of sitostanol and campestanol, are highly elevated. The high levels of saturated stenoles are most likely due to their more effective endogenous synthesis rather than more effective absorption (10, 27). If left untreated, sitosterolemia easily leads at early age to xanthomatosis and coronary heart disease. For people with this disease, an administration of sterols of unsaturated plants in amounts greater than what is normally present in food can lead to dangerous health effects. Lees and Lees (25) tested the effects of three different sitosterol preparations on lipid and plasma lipoprotein concentrations. One of the preparations was Cytellin, a commercial preparation (Eli Lilly Co., USA) that contained 60-65% of sitosterol and 35-40% of other sterols, mainly campesterol an average dose of 18 g / day divided into three doses resulted in an average 10.5% drop in total plasma cholesterol and a 15% drop in LDL-cholesterol. Nevertheless, has been shown when only traces of plant sterols including campesterol are normally detected in plasma (10, 33), the plasma concentration of campesterolles varied from 4 to 21 mg / dl in the subjects tested by Lees and Lees (25). In the discussion the authors stated very strongly that because the atherogenicity of campesterol is unknown, the use of a sitosterol preparation with a relatively high campesterol content is similar to the Cytellin preparation used in their study can not be recommended. In addition, Lees et al. (26) studied the effectiveness of plant sterols of soybean oil and bait oil by lowering the level of cholesterol in the blood. They used two different physical forms of each plant sterol, namely a suspension and a powder. The soy sterol consisted of 60-65% of sitosterol and 35% of campesterol and a daily dose of an average of 18g of sterols per day (scale 9-24g) was given in three equal doses. A sterol preparation of bait oil with only about 5% campesterol was used in this study. A daily dose of 3 grams of both sterol preparations of bait oil (powder and suspension) was tested. Additionally, a 6 gram dose of the sterol suspension of bait oil was tested. Soy sterol in both physical forms and sterol of bait oil reduced the plasma cholesterol content by an average of 12% (26). However, the relatively high absorption capacity of cam pesterol shown above was also shown in this study. In the 5 patients tested plasma campesterol levels ranged from 5 to 21 mg / dl (mean 16 mg / dl). Again, even if the effect that decreases the cholesterol of soy sterol is significant, the authors did not recommend its use as an agent that lowers cholesterol. In contrast, pharmaceutical plant sterol preparations should contain a minimum of campesterol and a maximum of sitosterol. Based on the two studies cited above, it can be concluded that the use of vegetable oil-based sterols such as soy sterol was not highly recommended. Sterols from saturated plants such as methanol and campestanol are present in most vegetable oils only in trace amounts. However, bait oil sterols contain 10-15% of sitostanol, the saturated form of sitosterol. Sitosethanol can also be formed by hydrogenation of the double bond in sitosterol. In recent studies with experimental animals and humans, it has been proven that sitostanol is more effective as a cholesterol-lowering agent than sitosterol (8, 16, 17, 18, 19, 36). An additional advantage of sitostanol is that it is virtually nonabsorbable. Several studies (eg, 9, 16, 17, 21) have shown that sitostanol is practically nonabsorbable while small amounts (<5%) of its unsaturated form of sitosterol (33) can be absorbed. Similarly, in an in vitro study Amstrong and Carey (6) also showed that cholestanol, a saturated form of cholesterol, was more hydrophobic and less absorbable than cholesterol. When the sitostanol is made by hydrogenation of most of the sources of usual plant sterols, another saturated plant sterol is formed, namely campestanol, from the campesterol. Even recently, relatively little has been known about the absorption capacity and possible hypocholesterolemic effect of this stanol. Based on the data cited above stating that saturated sterols are less absorbable than their unsaturated forms, it could be hypothesized that campestanol could be virtually unabsorbable. To study the absorption capacity of different sterols from Heinemann and other plants (20), they compared the intestinal absorption of cholesterol with campesterol, sitosterol, stigmaterol and also low concentrations of sitostanol and campestanol in humans by means of intestinal perfusion technique. The results showed that the absorption regime of the different plant sterols varied between different plant sterols, with an average of 4.2% of sitosterol, 4.8% of stigmaterol, 9.6% of campesterol and 12.5% of campestanol. The great variation between the absorption efficiency in the ten male subjects was detected. Therefore, according to Heinemann and others (20) it was found that campestanol is absorbed more efficiently than its campesterol in its unsaturated form. This again is the assumption based on studies cited above that showed that saturated sterols (sitostanol, cholestanol) may be less absorbable than unsaturated sterols (sitosterol, cholesterol). The reason for this remains unclear. Heinemann and others (20) speculated that, although the reason for this conflict has no conflicting result, it could be that the study of Amstrong and Carey (6) was done with in vitro conditions and that the theory of hydrophobicity is a main factor in micellar binding and / or absorption could not be relevant under in vivo conditions. However, this speculation does not explain the fact that several studies have been conducted that have shown the poorer absorption capacity of sitostanol compared with that of sytosterol under in vivo conditions. Therefore, the results of Heinemann and others (20) that are in conflict with previous results, remained unexplained by the authors. Sugano et al. (34) studied the hypocholesterolemic activity of maize sterols (composition: 31% campesterol, 4% stigmaterol and 65% sitosterol) and corn stems (composition: 31% campestanol and 69% sitostanol) obtained by hydrogenation of the corn oil sterol mixture. Two experiments in rats have been carried out. Both sterol and stanol showed hypocholesterolemic effects at the level of 0.5-1% of the diet when cholesterol was ingested (1% in the diet). In the first experiment, no significant difference was observed in the hypocholesterolemic effect of phytosterols and phytostanols. However, in the second experiment, at the same dietary levels, the phytostanols showed considerably greater capacity to decrease the concentration of plasma cholesterol than the phytosterols (statistically significant at p <0.02). However, rats fed the 1.0% stanol diet had significantly lower plasma cholesterol levels (p <0.02) than animals fed the cholesterol free diet. This was not observed in rats fed the 1.0% sterol diet. Sugano et al. (34) did not study the difference in the hypocholesterolemic effect between mixtures of stanols with a high content of sitostanol and a low content of campestanol (estero! Based on bait oil) and stanol mixtures with a substantially higher level. of campestanol (sterol based on vegetable oil). They compared the hypocholesterolemic effect of a mixture of unsaturated sterol with the corresponding saturated stanol mixture. Subsequent studies by this group of researchers have focused on the effect that specifically decreases sitoestanol cholesterol and compared it with sitoesterol (21)., 22, 23, 35). In fact, in a later publication (23) it refers to the phytostanol study mentioned above (34) mentioning only the hypocholesterolemic effect of β-sitostanol compared to β-sitosterol out discussing any hypocholesterolemic effect of saturated sterols (including campestanol) compared to unsaturated sterols. In the last studies mentioned above, sterol mixtures have been used the normal composition of hydrogenated bait oil sterols a high content of sitostanol (>90%). Miettinen and Vanhanen (30) have shown that sito-stanoi in the form of a fatty acid ester is more effective than free sitostanol in decreasing serum cholesterol levels. The latest studies have shown that the use of sitoestanol esters as part of the daily diet is an effective way to reduce the concentrations of total serum and LDL-cholesterol (13, 14, 15, 31, 37, 38). The benefit of using stanol esters instead of stanol free is also that stanol esters are fat soluble and therefore can be easily incorporated into a wide variety of foods without changing the taste, taste or physical behavior of the final product. The method for the preparation of fatty acid esters of sitostanol and the use of stanol esters soluble in foods has been described in the Patent of E. U.A. No. 5,502, 045 (2), incorporated herein by reference. Straub (3) suggests the use of saturated steels (sitostanol, clionaestanol, 22,23-dihydrobrasicastone, campestanol and mixtures thereof) in a method to form a food additive composition in which the stems are mixed with an edible solubilizing agent. , an effective amount of a suitable antioxidant and an effective amount of a suitable dispersant. It is intended that these food additives reduce the absorption of cholesterol from foods and beverages containing cholesterol, e.g. , meat, egg and dairy products. However, in this patent there are no data that show any clinical effect nor does the absorption of dietary sterols appear. Eugster et al. (1) teach the use of small amounts of sterols, their fatty acid esters and glycosides for the treatment of tumors. The preparation methods proposed by Eugster and others involve dangerous chemical reagents such as N, N'-carbonyl-diimidazoi, thionyl chloride and solvents similar to tetrahydrofuran, benzene, chloroform or dimethylformamide. Eugster and others comment on the possible use of these substances as dietetic foods and as food additives, but they do not present data on hypocholesterolemic effects or claim coverage for such use. From the description of Eugster and others, it is difficult to obtain a clear picture of how the final product is purified to produce a sufficiently pure sterol ester in large quantities sufficient to be used as a food component. The only purification process referred to is thin layer chromatography and high performance liquid chromatography. If this is the case, the preparation method referred to in the Eugster and others patent is limited to only small quantities. The Patent of E. U.A. No. 3,751, 569 (4) describes the addition of fatty acids from plant sterols to cook the oil in order to lower serum cholesterol levels in man. The purposes of the patent, for use in the esterification of free sterols, a method that in no case meets the requirement of the preparation of a food grade product. According to the patent, perchloric acid acting as a catalyst was carried out between a free sterol and a fatty acid anhydride. The catalyst and reagent used can not be accepted in feeding processes. In addition, the patent refers to fatty acid esters only from sterols of native plants.

The method proposed in German Patent DE 22 48 921 (5) for the esterification of sterols present in oils and fats by a chemical interesterification technique complies with the criteria of the food processes. In this patent, the free sterol and an excess of the fatty acid esters are added to a mixture of oil or fat, whereby the mixture of whole fats is then interesterified by a commonly known interesterification technique. The resulting fat blend virtually all free sterols have been converted to fatty acid esters. The purpose of this is to protect the sterols in vegetable and animal oils against possible changes during the process. The previous data show that campesterol, one of the sterols of main plants, is relatively efficiently absorbed. Therefore, it has been recommended that only mixtures of plant sterols with a minimum campesterol content can be used. This has led in practice to the use of mixtures of sterols such as bait oil sterols with a high content of sitosterol. The majority of tin mill work has turned sitosethanol alone. The study by Heinemann and others (20) showing campestanol, the saturated form of campesterol, is more easily absorbed than campesterol or sitosterol (12.5%, 9.6% and 4.2% respectively) has led to a "consensus" that mixtures of saturated sterols with "high" levels of campesterol are not safe due to the absorption of campestanol.

A clear evidence of this is that all the clinical studies covering the use of tinols (sitostanol) have been based on mixtures of sterols with a high level of sitostanol and a low level of campestanol. It is an established fact from many studies (v.gr., 8, 17, 18, 19, 23, 36), that the sitostanol, the saturated form of sitosterol, is more effective than the corresponding unsaturated sitosterol, to reduce the level of cholesterol in the blood. The saturated sterols are additionally absorbed in very limited amounts, which makes the use of saturated sterols a safe means to reduce cholesterol on a population basis. Unsaturated sterols, especially campesterol, are absorbed in amounts high enough to make strong recommendations against the use of sterols mixtures with high levels of campesterol (eg, mixtures of sterols based on vegetable oils) (25, 26). . Consequently, there has been a strong prejudice against the use of campestanol in any substantial amount as a substance that will be added to foods and this has severely limited the spectrum of phytosterol containing raw materials towards them that contain a relatively lower amount of campesterol and its saturated form, campestanol. Brief Description of the Invention This invention relates to plant stanols compositions containing sitostanol as a major component but with substantial amounts of campestanol, either in free form or esterified as fatty acid esters to lower serum cholesterol level in blood. The invention further relates to the use of stanol compositions containing sitostanol as the main component but also substantial amounts of campestanol or fatty acid esters thereof in edible products as a dietary component to lower serum cholesterol levels in the blood . The aim of the present invention is to extend the spectrum of raw materials of plants useful in the preparation of substances for edible products especially edible oils and fats and foods containing fats that are intended to control the serum cholesterol levels of the blood. The invention allows the use of raw materials for these plant oils and fats which contain in addition to the sitosterol also a substantial amount of campesterol. Suitable raw materials for use in the preparation of the compositions of the present invention are v. gr., corn, soybeans and rapeseed but also other plants with a composition of high potassiumterol in campesterol can be used. The novel composition of the present invention and especially its esterified form, can be incorporated into food substances such as cooking oils, margarines, butter, mayonnaise, salad dressings, shortenings, cheeses (including immature and immature cheeses) and other foods containing fats The composition of the present invention can also be consumed as such. Detailed Description of the Invention In accordance with the present invention, the plant stanol composition, in addition to its main component, contains sitostanol, also a substantial amount of at least 10% campestanol. The composition preferably contains as much as 20% a 40% and more preferably from 25% to 35%, v. gr. , approximately 30% campestanol or its fatty acid ester when the composition has been esterified to make it lipophilic. During this specification all percentages are given by weight, unless otherwise specified. In this specification the numbers in brackets refer to publications mentioned in the Reference List. The data obtained surprisingly and against the prevailing prejudice show that a mixture of hydrogenated stanol contains sitostanol as the main component but with substantial amounts of campestanol at least it is as effective as a mixture of stanol containing more than 90% of sitostanol and one level low of campestanol, indicating that campestanol is at least as effective in reducing cholesterol absorption as sitostanol. In addition, the data from serum sterol analyzes in blood, clearly show that campestanol remains virtually unabsorbed, with the serum content in blood being approximately 40% smaller than that of sitostanol. Therefore, a mixture of stanol containing sitostanol as a major component, but with substantial amounts of campestanol should be considered as safe as a conventional sterol-based bait based on tin mixture. These data are a strong contrast to current opinion regarding the efficacy and safety of stanol mixtures with high amounts of campestanol (see 20, 27, 34). The Patent of E. U.A. No. 5,502, 045 (2) showed that fatty acid esters of sitostanol are more effective in reducing the level of cholesterol in the blood than free sitostanol. Subsequent studies have clearly confirmed the cholesterol-lowering effect of a margarine containing fatty acid esters of fat-soluble sitostanol (e.g., 31). The use of stanol fatty acid esters instead of free stanols is crucial for an extensive use of these in various food products containing fats because only stanol fatty acid esters are soluble in edible oils and fats in amounts high enough to reach effective levels to reduce the absorption of both dietary and bile cholesterol from the digestive tract. The solubility of stanol esters in edible oils and fats is as high as 35-40%, wherein the solubility of free sterols in edible oils and fats is limited to a maximum of 2 weight percent only at a temperature of 21 °. C (24). Higher amounts could be incorporated using different surfactants, solubilizing or dispersing agents, but even the use of these substances does not ensure the solubility of fats. The use of the above substances is usually restricted or prohibited by law. In addition, free sterols at a level of 1% will affect the physical properties of the fat or oil, causing changes in the structure and physical behavior of the product. This is not the case when the stanol fatty acid esters are used because the physical properties of the fat blend can be easily modified by altering the fatty acid composition in the mixture. It is obvious that stanol fatty acid esters can easily be incorporated into other foods than margarines and spreads as described in this invention. The Patent of E. U.A. No. 5, 502, 045 (2) gives additional examples of possible uses, however, it is obvious to those skilled in the art that stanol fatty acid esters can be added to a wide variety of foods, especially foods containing fats Many methods for preparing fatty acid esters from sterols have been proposed. The drawbacks of these methods are that almost all of them use reagents, which can not be accepted in the production of a product intended to be used as a macronutrient in foods. The use of toxic reagents such as thionyl chloride or fatty acid anhydride derivatives is common. The preferred method for preparing stanol fatty acid esters is described in the U.S. Patent. No. 5, 502, 045 (2, incorporated herein by reference). This procedure is based on the interesterification process widely used by the edible fats and oils industry. This esterification process deviates advantageously from the previous methods in which no other substances are used than the free stanol, a fatty acid ester or a mixture of fatty acid esters and an interesterification catalyst such as sodium ethylate. An important aspect of the method is that one of the reagents, the fatty acid ester is used in excess and functions as a solvent, solubilizing the stanol under the conditions used (vacuum 5-15 mm Hg). The reaction gives a mixture of fatty acid esters and stanol fatty acid esters. The stanol fatty acid ester can be easily concentrated in almost pure stanol fatty acid esters by vacuum distillation, which removes the excess of the fatty acid esters. Alternatively the mixture can be added as such to the final fat blend before the deodorization step is carried out. The stanols are found in small amounts in nature, eg. , in wheat, rye, corn and tritrícalo and therefore can be found in small quantities (1 1, 14) in the food product. Tinnels can be easily produced by hydrogenation of mixtures of natural sterols. Only mixtures of bait sterols with sufficiently high purity (sterol content >); 98%) that will be used as such for food use, were commercially available in 1996. Plant sterols with substantial amounts of campesterol such as vegetable oil based on sterol mixtures can be obtained eg as a byproduct of production of tocopherol of vegetable oil distillates. The plant sterols obtained can be converted into tin steels by previously known hydrogenation techniques such as those based on the use of Pd / C catalyst in organic solvents (7, incorporated herein by reference). It is obvious to those skilled in the art that a wide variety of Pd catalysts and solvents can be used to carry out hydrogenation, which when done under optimized conditions leaves only small amounts of unconverted unsaturated sterols while the formation of the Tins and stents of normal dehydroxylated byproducts remain at a low level (<1.5%). The present invention compares the hypocholestecholemic effect of a mixture of tinols containing a high level of sitostanol which is generally considered by those skilled in the art to be the most effective plant sterol to reduce cholesterol absorption and therefore the levels of serum cholesterol with a mixture of stanol, contain a substantial amount of campestanol. In this specification, the first part has reported the hypocholesterolemic effects of tinols based on vegetable oils in humans. This invention is the first to show that a mixture of stanol with a substantial amount of campestanol (more than 10% and preferably about 30%) is at least as effective as mixtures of stanol with high levels of sitostanol. In addition, the results of the present study clearly indicate that campestanol, on the contrary to what has been reported by Heinemann and others (20), is virtually non-absorbed. Clinical Studies To study the hypocholesterolemic effects of stanol ester margarines from vegetable oil and stanol ester from bait oil, a double-blind, 5-week crossover study was designed with a washout period of 2 weeks. The test disposition of the study was as follows: Provision of proof of the intervention study The numbers 1-6 indicate that the blood samples recovered in the homemade diet (1, 2), after the first intervention period (3, 4) and after the second intervention period (5, 6). EV = stanol ester margarine based on vegetable oil, EC = stanol ester margarine based on bait oil.

Group 1. n = 12 1 2- ™ '"" * r 5 - 6 EV \ VE \ / Group 2. n = 12 / X \ -3 4 -5 * "' 6 EC Homemade Diet Period of period of intervention intervention intervention wash 5 weeks 2 weeks 5 weeks Twenty-four healthy, free-living, voluntary women with a moderate cholesterol level (average 6.12 + 0.16 mmol) consumed approximately 25 g per day (one barrel of 250 g / 10 days) of the test margarines as part of the daily diet in a random order. The serum lipids (total cholesterol, LDL-cholesterol, HDL-cholesterol and triglycerides) and serum sterol contents were measured in the domestic diet and at the end of each trial period. Blood samples were taken twice, a week apart on the homemade diet and at the end of the trial margarine period. The serum lipid values obtained are shown in Table 1 below. Table 1. Concentrations of serum lipids (mmol / l, mean + SE) during the homemade diet and after the five-week treatment with vegetable oil stanol ester margarine (EV) and stanol ester margarine from bait oil (EC), (n = 24) * p < 0.05 or less Both test margarines resulted in favorable serum lipid changes. The reduction in LDL-cholesterol values and the increase in H DL-cholesterol values were statistically significant (p < 0.05 or less). In addition, the sterol ester based on vegetable oil also resulted in a statistically significant reduction of total cholesterol. The reduction obtained from total cholesterol and LDL-cholesterol was higher with the stanol ester margarine based on vegetable oil compared to the stanol ester margarine based on bait oil. No changes in triglyceride levels were obtained. The serum lipid results obtained indicate that a stanol ester margarine from vegetable oil containing a substantial amount of campestanol in its stanol fraction can be even more effective than the stanol ester margarine from bait oil. Bait oil stanol ester margarine in previous studies (14, 15, 31) showed effective hypocholesterolemic effects. Therefore, based on the cross-over design of this study, it can be concluded that vegetable oil-based stems are at least shown as effective hypocholesterolemic effects as tinfoils based on bait oil. Serum sterol concentrations were quantified by gas-liquid chromatography according to a previously published method (29, incorporated herein by reference). The means of two measurements of serum lipids from the blood samples taken in each period were calculated. Data on sterol concentrations of average serum plants in the homemade diet and after each trial period and the average changes observed in these concentrations are presented in the following Tables 2 and 3. Table 2. Sterol concentrations of plants in serum (average + SE, μg / dl) during the homemade diet and after each intervention period (n = 24). Stanol ester margarine based on vegetable oil, EC = stanol ester margarine based on bait oil 'p < 0.05 or less Table 3. Mean changes (± SE) in sterol concentrations of serum plants (μg / dl), (n = 24). EV = stanol ester margarine based on vegetable oil, EC = stanol ester margarine based on bait oil, DC = homemade diet. íp < 0.05 or less Both test margarines significantly decreased the serum campesterol and serum sitosterol levels. The serum concentration of campesterol is known to reflect the absorption of intestinal cholesterol in humans (29, 39). Therefore, the lower the value of campesterol, the lower the percentage of intestinal cholesterol absorbed. The marked decreases in serum campesterol levels (25-28%) during the study periods indicate that stanol ester margarines decreased the intestinal absorption of cholesteroi. further, no differences in the concentration of sitostanol in serum can be observed while the concentration of serum campestanol after the stanol period of vegetable oil was significantly higher than the homemade diet and after the stanol ester period of bait oil. However, the absolute concentration of campestanol was only about 63% of that of sitostanol, which is generally considered to be virtually unabsorbable. This low serum concentration of campestanol clearly indicates that the absorption of campestanol is very limited, which is in conflict with the results presented by Heinemann and others (20). Therefore, because stanol mixtures containing high levels of sitostanol are considered safe for human ingestion. Stanol mixtures containing substantial amounts of campestanol should be considered as equally safe based on the fact that campestanol is virtually non-absorbable as sitostanol. The preparation of the stanol ester composition of the invention and the margarines used in the above clinical studies are described in detail in the following working examples. Example 1: Hydrogenation of sterol mixtures A commercially available sterol mixture obtained from vegetable oil distillate (composition: 2.7% brasicaesterol, 26.7% campesterol, 18.4% stigmaterol, 49.1% sterolol and 2.9% sitostanol) was hydrogenated in a reactor. pilot scale (25 liters). 26 g of fibrous Pd catalyst (Smop-20; content of Pd 10% by weight, Smoptech, Turku, Finland), 26 g of distilled water for the activation of the catalyst and 1 1 .7 kg. of propanol was fed into the reactor. The reactor was rinsed with nitrogen and the activation of the catalyst was carried out under hydrogen gas at a pressure of 1 bar and at a temperature of 65 ° C for 30 minutes. After activation the mixture was cooled to 40 ° C, after which 1.3 kg. of the sterol mixture was added. The propanol sterol mixture was heated under a nitrogen atmosphere at 65 ° C, after which the nitrogen was displaced by the hydrogen. After that, a step rinsing with hydrogen was carried out, the hydrogenation reaction was carried out at a hydrogen pressure of 1 bar. The normal conversion time is approximately 120 minutes. The conversion can be easily monitored by taking aliquots, which were analyzed by CLAR. The hydrogen pressure was lowered and the reactor rinsed with nitrogen. The fibrous catalyst was filtered under nitrogen pressure. The mixture of propanol stanol was allowed to crystallize overnight at 10 ° C after which the stanol crystals were filtered under vacuum and the cake was washed with 0.5 kg of cold propanol. The obtained stanol mixture was dried at 60 ° C in a vacuum cup. The yield was 75% and the composition of the stanol mixture obtained was the following according to capillary CG analysis: campesterol 0.2%, campestanol 28.9%, stigmaterol 0.1%, sitoesterol 0.2%, sitostanol 70.1%. it should be noted that the brassicaterol was hydrogenated in 24β-methyl cholestanol, an epimer of campestanol, but since it appears in the same peak with ordinary capillary gas chromatographic procedures that are not able to separate according to chirality, it is usually calculate as campestanol. Based on the initial sterol mixture the content of 24β-methyl cholestanol should be 1.7%. Example 2. Preparation of stanol fatty acid esters A mixture of stanol fatty acid esters was prepared on a pilot scale. 6 kg. of stanol obtained by combining several batches obtained by the hydrogenation process given in Example 1 was dried overnight at 60 ° C and esterified with a mixture of methyl ester of raquese oil of low erucic acid of 8.6 kg. The sterol composition of the stanol mixtures used were the following: Campesterol 0.4%, campestanol (+ 24β-methyl cholestanol) 29.7%, stigmaterol 0.1%, sitosterol 0.4% and sitostanol 68.0%. the stanol content of the mixture was 98.2%. the esterification was carried out in the following manner. A mixture of tin steels and fatty acid methyl ester with low erucic rapeseed oil was heated in a vessel in a reactor at 90-120 ° C under a vacuum of 5-15 mmHg. After drying for 1 hour, 21 g of sodium ethylate was added and the reaction was continued for about 2 hours. The catalyst was destroyed by the addition of 30% water (by weight) at 90 ° C. After phase separation, the water phase was removed and a second wash was carried out. After separation of the water phase, the oil phase was dried under vacuum at 95 ° C with a stirring effect of 200 rpm. The stanol fatty acid mixture was bleached slightly for 20 minutes at 30 mmHg at a temperature of 1 10 ° C with 1.0% bleaching earth (Tosü Optimum FF, Südchemie, Germany) under a stirring effect of 200 rpm. The bleaching earth was filtered and the mixture obtained from the fatty acid methyl esters and the stanol fatty acid esters can be added as such to the fat mixtures before the deodorization or the excess of methyl esters can be distilled under empty. Consequently, the mixture can be deodorized to obtain an unflavored stanol fatty acid ester mixture, which can be added as such to different food manufacturing processes. The conversion of the esterification process is normally of >99% measured by a fast CLAR method and the yield is on the 95% scale. Example 3: Production of Margarines for Clinical Studies 80% of margarine are stanol fatty acid esters of bait oil and stanol fatty acid esters based on vegetable oils were produced in a Gerstenberg pilot scale perfector & Aggregate 3 x 57. The stanol fatty acid esters of bait oil were obtained from the normal production of Benecol® margarine by Raision Margariini, Finland. A mixture of normal trans fatty acids free fats (composition: 30% non-hydrogenated interesterified vegetable fat and 70% LEAR liquid oil) to which stanol fatty acid mixtures were added was used. The stanol content of the final product to be 12 g / 100 g of product, which provided a daily consumption of 3 g of stanols at a use level of 25 g / day. The products were produced according to the following recipe: Fat blend including esters of 80% stanol fatty acids Water 19% Salt 0.5% Emulsifier, Dimodan BP sodium bicarbonate and citric acid as pH-regulating agents-carotene as agent Flavor coloring The margarines obtained were minced in 250 g polypropylene barrels, which were sealed by an aluminum sheet. The flavor and texture of the products were the same as commercial margarines. The stanol content of the stanol margarine of bait oil was 12.7 g / 100 g product and the vegetable oil based on stanol margarine 12.6 g / 100 g product. The sterol composition of the two products was as follows: Stanol margarine Stanol margarine based on vegetable oil based bait oil Brassensterol 0.3% 0.4% Campesterol 2.2% 2.4% Campestanol 7.5% 27.6% Sitosterol 7.4% 4.2% Sistoestanol 82.5% 63.8% Others 0.1% 1.6% List of References Patent Specification of E.U.A. Ref. No. 1. Eugster C, Eugster C, Haldemann W, Rivara G. Sterols, their fatty acid esters and glocosides; processes for their preparation; spontaneously dispersible agents containing these compounds, and their use for treatment of tumors, 1993. Patent of E.U.A. No. 5,270,041. 2. Miettinen TA, Vanhanen H, Wester I. Use of stanol fatty acid ester for reducing blood serum level. 1996. Patent of E.U.A. No. 5,502,045. 3. CD Straub. Stanols to reduce cholesterol absorption from foods and methods of preparation and use thereof. 1993. Patent of E.U.A. No. 5,244,887. 4. Clear cooking and salad oils having hypocholesterolemic properties. 1973. Patent of E.U.A. No. 3,751,569. Another Patent Specification 5. Baltes J, Merkle R. Verfahren zur Herstellung eines Gemisches aus pflanzlichen und tierischen Olen bzw. Fetten und Fettsáuresternestern. German Patent DE 2248921. Other publications 6. Amstrong MJ, Carey MC. Thermodynamic and molecular determinants of sterol solubilities in bile salt micelles. J Lipid Res 1987; 28: 1144-1155.

Augustine RL, Reardon Jr. EJ 1969. The paladium Catalyzed hydrogenation of cholesterol. Org Prep and Proce 1969; 1: 107-109. Becker M, Staab D, Von Bergmann K. Treatment of severe familial hypercholesterolemia in childhood with sitosterol and sitostanol. J. pediatr 1993; 122: 292-296. Czubayko F, Beumers B, Lemmsfuss S, Lütjohann D, von Bergmann K. A Simplified micro-method for quantification of fecal excretion of neutral and acidic sterols for autopatient studies in humans. J Lipid Res 1991; 32: 1861-1867. Dayal B, Tnt GS, Batta AK, Speck J, Khachadurian AK, Shefer S, Salen G. Identification of 5-a stanols in patients with sitosterolemia anddxanthomatosis: stereochemistry of the protonolysis of steroidal organoboranes. Steroids 1982; 40: 233-243. Dutta PC, Appelqvist LÁ. Saturated sterols (stanols) in unhydrogenated and hydrogenated edibie vegetable oils and in cereal lipds. J Sci Food Agrie 1996; 71: 383-391. Grundy SM, Mok HYI. Effects of low dose phytosterols on cholesterol absorption in man. In: Greten H (Ed.) Lipoprotein metabolism. Springer-Verlag, Berlin, Haidelberg. New York, 1976: 112-118. Gylling H, Miettinen TA, Lowering serum cholesterol by dietary systostanol, and associated with reduced absuption and synthesis of cholesterol and decreased transport of LDL apoprotein B in men with type 11 diabetes. In: Gotto Jr AM, Mancini M, Richter WO, Schwandt P (Eds.) Treatment of severe dyslipoproteinemia in the prevention of coronary heart disease. 4th Int Symp Munich 1992, Karger, Basel, 1993: 57-59. 14. Gylling H, Miettinen TA. Serum cholesterol and cholesterol lipoprotein metabolism in hypercholesterolemic NIDDM patients before and during sitostanol ester-margarine treatment. Diabetoiogia 1994; 37: 773-780. 15. Gylling H, Sumes MA, Miettinen TA. Sitostanol ester margarine in dietary treatment of children with familial hypercholesterolemia. J Lipid Res 1995; 36: 1807-1912. 16. Hassan AS. Rampone AJ. Intestinal absroption and lympatic transport of cholesterol and ß-sistostanol in the rat. J Lipid Res 1979; 20: 646-653. 17. Heinemann T, Leiss O, von Bergmann K. Effect of low-dose sitostanol on serum, in patients with hypercholesterolemia. Atherosclerosis 1986; 61: 219-223. 18. Heinemann T, Pietruck B, Kullack-Ublick G, von Bergmann K Comparison of sitosterol and sitostanol on inhibition of intestinal absorption. Agents Actions (Suppl) 1988; 26: 117-122. 19. Heinemann T, Kullak-Ublick G-A, Pietruck B, von Bergmann K. Mechanisms of action of plan sterols on inhibition of Absroption. Eur J Clin Pharmacol 1991; 40: 59-63.

. Heinemann T, Axtmann G, von Bergmann K. Comparison of intestinal absuption of cholesterol with differnt plant sterols in man. Eur J Clin Invest 1993; 23: 823-381. twenty-one . Ikeda I, Sugano M. Comparison of absuption and metabolism of β-sistosterol and β-sitostanol in rats. Atherosclerosis 1978; 30: 227-237. 22. Ikeda I, Tanabe Y, Sugano M. Effects of sistosterol and sistoanol on micellar solubility of cholesterol. J N utr Sci Vitaminol 1989; 35: 361-369. 23. Ikeda I, Kawasaki A, Samezima K, Sugano M. Antihypercholesterolemic activity of β-sitostanol in rabbits. J Nutr Sci Vitaminol 1981; 27: 243-251. 24. Jandacek RJ, Webb MR, Mattson FH. Effect of an aqueous phase on the solubility of cholesterol in an oil phase. J Lipid Res 1977; 18: 203-210. 25. Lees RS, Lees AM, Effects of sitosterol therapy on plasma lipid and lipoprotein concentrations. I n: Greten H (Ed.) Lipoprotein Metabolism. Springer-Verlag. Berlin, Haidelberg, New York, 1976: 1 19-124. 26. Lees AM, Mok HYI, Lees RS, McCIuskey MA, Grundy SM. Plant Sterols as lowering agents: clinical triais in patients with hypercholesterolemia and studies of balance. Atherosclerosis 1977; 28: 325-338. 7. Ling WH, Jones PJH. Minireview dietary phytosterols: A review of metabolism. Benefits and side effects. Life Sciences 1995; 57: 1995-206. 8. Mattson FH, Grundy SM, Crouse JR. Optimizing the effect of plant sterols on cholesterol absorption in man. Am J Clint Nutr 1982; 35: 697-700. 9. Miettinene TA, Koivisto P. Non-cholesterol sterols and bile acid production in hypercholesterolaemic patients with loyal bypass. In: Paumgarter G, Stiehl A, Gerok W (Eds.). Bile acid and concentration in health and diseade. MTP Press, Boston 1983: 183-187. 30. Miettinen TA, Vanhanen H, Dietary sitostanol related to absorption, synthesis and serum level of cholesterol in different apolipoprotein E phenotypes. Atherosclerosis 1994; 105: 217-226. 31. Miettinen TA, Puska P, Gylling H, Vanhanenen H, Vartiainen E. Reduction of serum cholesterol with sitostanol-ester margarine in a midly hypercholesterolemic population. New Engl J Med 1995; 333: 1308-1312. 32. Pollak OJ. Effect of plant sterols on serum lipids and atherosclerosis. Pharmac Ther 1985; 31: 177-208. 33. Salen G. Ahrens Jr. EH, Grundy SM. Metabolism of ß-sítosterol in man. J Clin Invest 1970, 49: 952-967. J Nutr Sci Vitaminol 1981; 27: 243-251. 34. Sugano M, Kamo F, Ikeda Y, Morioka H. Lipid-lowering activity of phytostanols in rats. Atherosclerosis 1976; 24: 301-309.

Sugano M, Morioka H, Ikeda I. A comparison of hypocholesterolemica activity of ß-sistosterol and ß-sítostanol in rats. J N utr 1977; 107: 201 1 -2019. Vanhanenen HT, Mielttineen TA. Effects of unsaturated and saturated dietary plant sterols on their serum contents. Clin Chim Acta 1992; 205: 97-107. Vanhanen HT, Blomqvist S, Enholm C, Hyvonene M, Jauhiainen M, Torstila I, Miettineen TA. Serum cholesterol, precursors, and plant sterols in hypercholesterolemics subjects with different apoE phenotypes during dietary sitostanol ester teatment. J Lipid Res 1993; 34: 1535-1544. Vanhanen HT, Kajander J. Lehtovirta H, Miettineen TA. Serum levéis, absroption efficiency, faecal elimination and synthesis of cholesterol during increasing doses of dietary sitostanol esteres in hypercholesterolaemic subjects. Clin Sci 1994; 87: 61 -67. Tilvis RS, M iettineen TA. Serum plant sterols and their relation to absroption. Am J Clin Nutr 1986; 43: 92-97.

Claims (10)

1. RETIREMENT D ISSION 1. A composition of plant stanols for use as a substance that lowers serum cholesterol level and which comprises sitostanol, the composition further comprises at least 10% campestanol, as long as the composition does not contain 31% campestanol and 69% of sitostanol.
2. 2. The composition of claim 1, comprising from 20% to 40%, preferably from 25% to 35% campestanol.
3. 3. The composition of claim 1 or 2, comprising from 50% to 80% of sitostanol.
4. 4. A plant stanol fatty acid ester composition comprising a fatty acid ester of sitostanol for use as a substance that reduces serum cholesterol level, the composition further comprising at least 10% of an ester of fatty acid of campestanol.
5. The composition of claim 4, comprising from 20% to 40%, preferably from 25% to 35%, v. gr. , approximately 30% of the fatty acid ester of campestanol.
6. 6. The composition of claim 4 or 5, comprising 50 to 80% fatty acid ester of sitostanol.
7. 7. The use of a composition of any of claims 1 to 6, as such or as part of the diet, e.g. , in foods that contain fats, which will be consumed to lower serum cholesterol levels.
8. 8. A food substance containing a stanol composition of plants containing sitosethanol or a fatty acid ester thereof effective to decrease the serum cholesterol levels, the composition further comprising a substantial amount of campestanol or a fatty acid ester of the same so that the weight ratio of campestanol or its fatty acid ester to sitostanol or its fatty acid ester is from 1: 9 to 4: 6, as long as the stanol composition is not composed of a composition of free stems containing 31% campestanol and 69% sitostanol.
9. 9. The food substance of Claim 8, wherein the ratio by weight is from 2: 8 to 3.5: 6.5.
10. 10. The use of a mixture of fistoesterol, which also comprises sitoesterol a substantial amount of campesterol, as a raw material to produce a composition that decreases the serum cholesterol level or a food substance according to any of claims 1 to 6, 8 and 9. RESU MEN A stanol composition containing in addition to sitostanol as the main component, also a substantial amount of at least 10% campestanol, has been found to effectively decrease serum cholesterol levels when incorporated into edible articles. By esterifying the composition it is especially useful in edible fats and oils and in foods containing fats.