HK1061955B - Method for dispersing plant sterol and a beverage containing nanometer-scale plant sterol - Google Patents

Description

Method for dispersing plant sterol and beverage containing plant sterol of nanometer grade

Technical Field

The present invention relates to a method for dispersing phytosterol for use in a beverage and a phytosterol dispersed beverage. More particularly, the present invention relates to a method for preparing a dispersion of phytosterols wherein the size of the phytosterol micelles formed is hundreds of nanometers or less, and to beverages containing the dispersion.

Background

Recently, excessive intake of cholesterol has been increasing, and as a result, diseases associated with cholesterol are becoming a big social problem. Especially easterns, whose lifestyle mimics westernization, including diet, have had more opportunities for consuming high cholesterol foods than ever before due to the fact that heat is faithful to instant foods and snacks. Once ingested, the cholesterol in these foods increases the cholesterol level in the blood and may be a major cause of cardiovascular disease, including hyperlipidemia, arteriosclerosis, arrhythmia, cardiac infarction, and the like.

By studying the metabolic processes of cholesterol, it has been shown that both endogenous and dietary cholesterol enters the small intestine and that about 50% of the cholesterol is absorbed by the intestine (Bosner, m.s., Ostlund, r.e., jr., osofossan, o., Grosklos, j., Fritschle, c., Lange, l.g. 1993). Based on this fact, the mechanism of action that prevents the absorption of cholesterol by the intestine is of particular interest to those who are working to find clues for the prevention and treatment of cholesterol-related diseases.

Phytosterols or phytosterols may be classified as sitosterol, campesterol and stigmasterol, while phytosterols or phytosterols include sitosterol and campesterol, which are collectively referred to herein as phytosterols for convenience.

Since the structure of phytosterols is very similar to that of cholesterol, phytosterols are known to inhibit intestinal absorption of cholesterol, thereby reducing serum cholesterol levels, as disclosed in U.S. patent No. 5,578,334. Phytosterols, as naturally occurring substances, are non-toxic and can be found in a wide spectrum of plants, such as soybeans, corn, trees, tall oil (tall oil), and the like. Using the function of phytosterols in inhibiting intestinal absorption of cholesterol, clinical trials have used phytosterols as therapeutic agents for the treatment of cardiovascular disease, coronary artery disease and hyperlipidemia (Atherosclerosis 28: 325-338).

Despite this useful function, phytosterols are difficult to use in food products due to their physical properties, i.e., poor solubility in water and oil. Thus, the general public has only limited uptake of phytosterols.

In order to increase the solubility of phytosterols, several researchers have synthesized various derivatives of phytosterols. For example, ester compounds of phytosterols have been developed which have excellent solubility in the oil phase (Mattson f.h., r.a.volpenhein and b.a.erickson, 1997). U.S. Pat. No. 5,502,045 discloses sitosterol fatty acid esters, which are prepared by esterification of sitosterol with fatty acids. This sitosterol fatty acid ester is reported to reduce the LVL-C content by up to 16% when used in a practical form in the oil phase (margarine).

Water-soluble and oil-soluble phytosterol derivatives as described in WO 99/15546 and WO 99/15547 are synthesized by linking water-or oil-soluble molecules to phytosterols or plant stanols via ester linkages.

However, some studies have shown that synthetic phytosterol derivatives with improved solubility have less potent inhibition of intestinal cholesterol absorption than natural phytosterols (Mattson et al, the American Journal of Clinical Nutrition 35: April 1982, p. 697-700). In particular, such oil-soluble derivatives have the disadvantage that a large amount of edible oil must also be absorbed simultaneously.

In addition to efforts to improve the solubility of phytosterols by synthesizing derivatives thereof, intensive research has been and continues to be conducted to improve the bioavailability of phytosterols.

Much work has been done on this area. For example, a pharmaceutical sitosterol dispersible powder for oral administration has been developed, which is prepared by mixing sitosterol, starch hydrolysate, silica and polyoxy-xylene (polyoxylene) sorbitan monostearate in a certain ratio, homogenizing, degassing, heat sterilizing and evaporating, as disclosed in U.S. Pat. No. 3,881,005.

U.S. Pat. No. 5,932,562 discloses an aqueous homogeneous micellar mixture of phytosterols, lecithin and lysolecithin, which is dried to a finely divided water-soluble powder. It is obtained by mixing together phytosterol, lecithin and lysolecithin in chloroform at a fixed molar ratio and then removing the chloroform. However, this patent also presents some problems. Wherein the total amount of emulsifier is far greater than the amount of phytosterol. The emulsifier lysolecithin is expensive. Worse still, the organic solvent used to form the micelles makes the water-soluble powder unsuitable for absorption.

Other water soluble phytosterols can also be found in U.S. patent nos. 6,054,144 and 6,110,502. According to these patents, water dispersible phytosterols are produced by mixing sitosterol or phytosterol, a monofunctional surfactant and a polyfunctional surfactant in water at a certain ratio and then drying the mixture. The production method is characterized in that the steps of homogenization and degassing are eliminated, and polyoxy xylene sorbitan monopalmitate and sorbitan monopalmitate are used as the monofunctional surfactant and the polyfunctional surfactant, respectively.

European patent publication No. 289,636 describes a method for producing stabilized emulsified or solubilized sterols by mixing a plant sterol with a polyol liquid containing sucrose fatty acid ester and/or polyglycerin fatty acid ester in a certain ratio and diluting the mixture with water. When this is used in beverages, the resulting particulate micelles of phytosterols are as large as tens of microns in size and taste like bristles. Furthermore, a disadvantage of such micro-agglomerated particles is that the beverage is not transparent.

Food ingredients that can be used to reduce cholesterol levels are disclosed in U.S. patent No. 6,190,720. This patent also describes a process for preparing a food ingredient by mixing one or more molten phytosterols with one or more fats and one or more emulsifiers to homogeneity and cooling the homogenate to about 60 ℃ with agitation to obtain a paste. The food ingredient can be used in oil-based foods such as salad dressing, margarine, etc. As expected, this food ingredient is virtually impossible to use in aqueous beverages because its dispersion stability is only obtained in fat.

European patent No. 0897671a1 relates to aqueous dispersions of phytosterols for use in spreads, dressings, milk, cheese and the like and to a process for their preparation which comprises mixing together under shear a molten high melting lipid, a non-sterol emulsifier and water, characterized in that the high melting lipid has an average size of 15 microns or less. Aqueous dispersions have the advantage of minimizing or eliminating saturated fats and trans fatty acids. However, micronization of high melting point lipids such as phytosterols must be performed. In addition, this dispersion cannot be used for aqueous beverages due to its low dispersion stability in water.

A cholesterol-lowering edible product can be found in WO 00/33669. According to the method of this patent, phytosterols are dissolved or mixed in a melt of food emulsifier, mixed with a protein-containing food such as milk or yogurt, homogenized, and then added to the food. The cholesterol-lowering edible product maintains its dispersion stability only in the presence of the protein-containing material, and does not maintain its stability in the absence of the protein-containing material. Therefore, it is difficult to use such cholesterol-lowering edible products in beverages that do not contain protein materials.

Us patent No. 6,267,963 relates to a phytosterol-emulsifier complex having a melting point at least 30 ℃ lower than the melting point of phytosterols, characterized in that due to its reduced melting point, the phytosterol-emulsifier is less likely to crystallize during or after food processing and can be incorporated into food products in an amount effective to reduce serum cholesterol levels, such that one does not need to adversely modify the texture of the food products when consuming these products. The complex prepared from phytosterol, emulsifier, neutral lipid such as triglyceride can be used in oil-based food. If no neutral lipids are present, sodium stearoyl lactylate may be used as an emulsifier. The use of sodium stearoyl lactylate is subject to legal restrictions. Furthermore, when such compounds are used in beverages, their characteristic unpleasant odor also requires masking processes.

DISCLOSURE OF THE INVENTION

In view of the above-mentioned problems, after intensive and thorough research into soluble forms of phytosterols, the present inventors have found that in the absence of other ingredients, phytosterols are heated together with an emulsifier so as to be brought into uniform contact with each other and fused to form fine micelles of small to nano-scale sizes under high-speed stirring and homogenization, thereby leading to the present invention. It was also found that in beverages, the nano-sized micelles are excellent in bioavailability, have no influence on the characteristic taste and flavor of the beverage, are used in almost all beverages regardless of the base and pH of the beverage, and improve the dispersion stability of the phytosterol micelles to prolong the life cycle of the beverage, ensuring long-term stability of the product.

It is therefore an object of the present invention to provide a method for dispersing phytosterols in an aqueous matrix in a convenient form which is suitable for use in beverages and which enhances the bioavailability of phytosterols, in addition to having no effect on the characteristic taste and aroma of the beverages used.

It is another object of the present invention to provide a beverage containing a dispersion of plant sterols which does not taste as bristled.

It is a further object of the present invention to provide an additive suitable for use in beverages which can be prepared by the process of the present invention.

According to one embodiment of the present invention, there is provided a method of dispersing phytosterols comprising the steps of: thermally melting a mixture of phytosterols and at least one emulsifier selected from the group consisting of sucrose fatty acid esters, sorbitan fatty acid esters and polyglycerine fatty acid esters at a temperature of 60-200 ℃; mixing the molten mixture with an aqueous beverage or an aqueous beverage containing an emulsifier; the mixture is stirred at high speed to obtain a beverage of a dispersion of phytosterols, whereby the phytosterols can be dispersed into particles of hundreds of nanometres in size.

According to another embodiment of the present invention, there is provided a method of dispersing phytosterols comprising the steps of: hot-melting a mixture of a plant sterol and an emulsifier selected from the group consisting of sucrose fatty acid esters, sorbitan fatty acid esters and polyglycerin fatty acid esters at a temperature of 60-200 ℃; mixing the molten mixture with an aqueous beverage or an aqueous beverage containing an emulsifier; and stirring and homogenizing the mixture at a high speed to obtain a plant sterol-dispersed beverage, whereby the plant sterol can be dispersed into particles of several hundred nanometers in size.

According to another embodiment of the present invention, there is provided a method of dispersing phytosterols comprising the steps of: thermally melting a mixture of phytosterols and at least one emulsifier selected from the group consisting of sucrose fatty acid esters, sorbitan fatty acid esters and polyglycerine fatty acid esters at a temperature of 60-200 ℃; cooling the melt, solidifying, grinding the solid to obtain a powder, and mixing the powder with an aqueous beverage or an aqueous beverage containing an emulsifier; and stirring the mixture at a high speed to obtain a plant sterol-dispersed beverage, whereby the plant sterol can be dispersed into particles of several hundred nanometers in size.

According to another embodiment of the present invention, there is provided a method of dispersing phytosterols comprising the steps of: thermally melting a mixture of phytosterols and at least one emulsifier selected from the group consisting of sucrose fatty acid esters, sorbitan fatty acid esters and polyglycerin fatty acid esters at a temperature of 60-200 ℃; cooling the melt, solidifying, grinding the solid to obtain a powder, and mixing the powder with an aqueous beverage or an aqueous beverage containing an emulsifier; and stirring and homogenizing the mixture at a high speed to obtain a plant sterol-dispersed beverage, whereby the plant sterol can be dispersed into particles of several hundred nanometers in size.

According to another embodiment of the present invention, there is provided a plant sterol-dispersed beverage prepared by one of the above-mentioned methods.

According to another embodiment of the present invention, there is provided an additive suitable for use in a beverage, characterized in that the additive is prepared as follows: a mixture of phytosterol and at least one emulsifier selected from sucrose fatty acid ester, sorbitan fatty acid ester and polyglycerin fatty acid ester is hot-melted at 60-200 deg.C to form particles having a size of several hundred nanometers or less when dispersed in an aqueous beverage.

Best Mode for Carrying Out The Invention

Naturally occurring phytosterols are structurally similar to cholesterol. In nature, many classes of phytosterols have been found, with sitosterol (sitosterol), campesterol (campesterol), stigmasterol (stigmasterol) and sitostanol (sitostanol) being greater than others. At normal intake levels, phytosterols have little effect on serum cholesterol levels, but large amounts of phytosterols inhibit the absorption of intestinal cholesterol and bile cholesterol because their structures are similar to that of cholesterol, thereby lowering serum cholesterol levels.

The effect of phytosterols on lowering serum cholesterol has been studied for a long time. These studies have shown that phytosterols can reduce total serum cholesterol levels by 0.5-26%, and particularly by 2-33% LDL-C levels, depending on the dosage, severity of the patient's symptoms and prescribed diet. The effect of the reduction of cholesterol content is also influenced by the sex, age and health of the patient and the form of administration (suspension, crystals, micelles, etc.).

It is now a generally accepted hypothesis that phytosterols effectively accumulate dietary cholesterol, which resides in the oil phase in the intestine and settles, thereby inhibiting the absorption of dietary cholesterol by the intestine. Thus, phytosterols are believed to be ineffective against cholesterol present in micellar form. In fact, Dietary Cholesterol has been reported to be more resistant to phytosterol Absorption than endogenous Cholesterol (Mattson, F.H., Volpenhein, R.A. and Erickson, B.A., Effect of plant Sterol Esters on the Absorption of Dietary Cholesterol, J.Nutr.1977; 107: 1139-. However, U.S. Pat. No. 5,932,562, which conducted one experiment with phytosterols, believes that micellar mixtures of phytosterols are very effective in reducing serum cholesterol levels even when applied in small amounts. It is believed that the micellar phase of phytosterols allows good cholesterol settling in the micellar phase and thus effectively inhibits intestinal absorption.

To date, the use of phytosterols to reduce blood cholesterol levels, particularly LDL-cholesterol, has led to the development of a number of commercial products which are sold in many countries such as Finland, the United kingdom, the United states and Odalia. They are mainly used as spreads or dressings, where phytosterols are bonded to fatty acids via ester bonds. These products are reported to be more effective in combination with statin drugs in hypercholesterolemic patients. However, this product contains a large amount of fat components so that it is also absorbed at the same time.

Many attempts have been made to provide phytosterols in the form of beverages. However, since phytosterols are slightly soluble in water, most beverage forms developed to date have the following disadvantages: poor bioavailability of phytosterols, lack of dose-balancing and the need for large amounts of solubilizers. Furthermore, the inhibition of cholesterol absorption achieved in the form of conventional aqueous matrices of phytosterols requires a significantly long time, since their availability in vivo is slow. Further, the dispersion stability of phytosterol is so poor that a drying process is also required to remove a solubilizing agent such as water for dispersing phytosterol, thereby obtaining powdery phytosterol. To avoid these problems, various solutions have been proposed, including grinding the phytosterols into a fine powder, adding at least two different additives, and using a high pressure homogenizer (5,000 psig).

As described above, various solubilizing agents and emulsifiers have been used to dissolve phytosterols, and it is reported in U.S. Pat. No. 5,932,562 that sitosterol can be easily dispersed by forming micelles from an organic solution of sitosterol. In general, this process and the resulting micellar mixtures are not suitable for beverages due to the large impact on the taste and aroma of the beverage. For example, large amounts of solubilizers or emulsifiers have a negative effect on the taste of the final edible product. In addition, organic solvents that are not normally suitable for use in food products may remain in the micelles.

The present invention thus relates to a method for dispersing phytosterols into micelles of several hundred nanometers in size, which improves the bioavailability of sparingly soluble phytosterols, has proper dose balance, and maximizes dispersion stability.

According to the present invention, the bioavailability of phytosterols is greatly improved, resulting in a reduction in effective doses. Furthermore, a completely clear dispersion of phytosterols was obtained, which had no effect on the taste and aroma of the desired beverage. The micellar particles of the present invention are so small that they are not bristled to the mouth after use. It is a further advantage of the present invention that such cholesterol-lowering beverages can be prepared without regard to their pH and composition.

The phytosterols useful in the present invention are selected from the group consisting of sitosterol, campesterol, stigmasterol, sitostanol, campestanol (campestanol) and mixtures thereof. In addition, other phytosterols may be used in the present invention.

Examples of the emulsifier which can disperse the plant sterol in the form of micelles having a size of several hundred nanometers or less according to the dispersion method of the present invention include sucrose fatty acid esters, sorbitan fatty acid esters and polyglycerol fatty acid esters. Emulsifiers other than sucrose fatty acid esters, sorbitan fatty acid esters, and polyglycerin fatty acid esters were found to produce a significant amount of particles having a size of 1 micron or greater, as determined by various experiments. In fact, other emulsifiers exhibited such low dispersion stability that precipitation occurred within three days after the dispersion of phytosterol. Therefore, emulsifiers other than sucrose fatty acid ester, sorbitan fatty acid ester and polyglycerin fatty acid ester emulsifiers are basically unsuitable for use in preparing beverages. Although sodium stearoyl lactylate is optimal in terms of dispersion stability, its use or amount is legally severely limited due to its lack of safety. In addition, the emulsifier has a characteristic unpleasant taste.

In contrast, the emulsifier used in the present invention disperses the phytosterol in a size of hundreds of nanometers or less. Moreover, the dispersion stability is so good that a 1% dispersion of phytosterol can be preserved for one year or more. Preferred among sucrose fatty acid esters are those having a Hydrophilic Lipophilic Balance (HLB) of 7 or more. Their HLB value is more preferably between 10 and 16. The sorbitan fatty acid ester preferably has an HLB value of between 5 and 10, more preferably between 7 and 10. As for the polyglycerin fatty acid ester, the HLB value is preferably between 10 and 20, and more preferably between 12 and 15. The particles obtained by using sorbitan fatty acid ester as emulsifier are smaller than those obtained by using sucrose fatty acid ester and polyglycerin fatty acid ester as other emulsifiers, and the particle size distribution is more uniform. In addition, these esters give off some unpleasant taste. In conclusion, sucrose fatty acid esters are most preferred.

In the dispersion of the present invention, the weight ratio of the phytosterol to the total emulsifier is in the range of 1: 0.01-10, and preferably in the range of 1: 0.2-2.0 (w/w). For example, if the weight ratio of the emulsifier to the phytosterol is less than 0.01, the emulsifier cannot be sufficiently emulsified to cause precipitation, and even if milky particles are formed, the particle size thereof is several tens of micrometers. On the other hand, if the weight ratio exceeds 10, the taste of the resulting beverage is that of an emulsifier, and the mouthfeel is too poor. As for the emulsifier mixed with the aqueous beverage, the weight ratio thereof is 0.8 or less (i.e., 80% by weight or less based on the weight of the emulsifier mixed with the plant sterol) as compared with the emulsifier mixed with the plant sterol, and preferably 0.5 or less (i.e., 80% by weight or less) by weight. A weight ratio of more than 0.8(w/w) (80% by weight) makes it difficult to form nanoparticles because the amount of emulsifier mixed with phytosterol is relatively low.

A wide variety of aqueous beverages can be used in the present invention, such as water, fruit juice beverages, soda water, milk, soy milk, cereal beverages, other popular beverages such as coffee, green tea, polyghace odoratum variant (var. odoratum) tea, and the like, and alcoholic beverages.

According to the invention, the phytosterols and the emulsifier are mixed at 60-200 ℃. The heating temperature of the mixture is preferably in the range of 120-150 ℃. When mixing is performed at below 60 ℃, the micelle particle size is tens to hundreds of micrometers, which is detrimental to taste and bioavailability. On the other hand, the mixing temperature is higher than 200 ℃ and the emulsifier is denatured, although the phytosterols are still stable at 250 ℃.

In principle, when phytosterols, a sparingly water-soluble substance, are emulsified in water in the presence of an emulsifier, only poor emulsification occurs, resulting in precipitation of the phytosterols onto particles ranging in size from tens to hundreds of microns. In the present invention, intensive research has been directed to maximizing emulsification of phytosterols, thereby obtaining micellar particles having a size of several hundred nanometers or less. As a result of intensive studies, it has been found that the emulsification can be maximized when a plant sterol and an emulsifier such as sucrose fatty acid ester, sorbitan fatty acid ester and polyglycerin fatty acid ester or a mixture of these emulsifiers are mixed to homogeneity. In order to mix the phytosterol and the emulsifier uniformly, the phytosterol was heated at a temperature near its melting point (sitosterol: about 140 ℃ C.; campesterol: about 157 ℃ C.; stigmasterol: about 170 ℃ C.) to turn the two components into liquid, and then mixed.

The aqueous beverage or the aqueous beverage containing the emulsifier is added to the hot mixture of phytosterols and emulsifier under high speed stirring. The emulsifier added to the aqueous beverage is preferably the same as the emulsifier mixed with the phytosterols. However, different emulsifiers can also be used if the two different emulsifiers are compatible with one another. The weight ratio of phytosterol to aqueous beverage is in the range of 1: 10 to 1: 10,000(w/w), and preferably in the range of 1: 10 to 1: 100 (w/w).

Examples of aqueous beverages useful in the present invention include water, fruit juice beverages, soda, milk, soy milk, cereal beverages, other popular beverages such as coffee, tea, etc., and alcoholic beverages, and are more preferred to water. In the case of using water, the dispersion of phytosterol obtained by the subsequent high-speed stirring and homogenization process may be further diluted with aqueous beverages such as water, fruit juice beverages, soda water, milk, soy milk, cereal beverages, other popular beverages, and alcoholic beverages to obtain the desired beverage containing phytosterol.

In order to ensure stable product quality and obtain particles with uniform size distribution, high-speed stirring and homogenizing processes are important in industry.

Alternatively, the phytosterol may be mixed with the emulsifier, heated near its melting point, the melt cooled to obtain a solid, and the solid ground into a powder. A beverage containing phytosterol is prepared by simply adding the powder to an aqueous beverage or a beverage containing an emulsifier. Additives in powder form have the advantage over liquid form that they are easy to handle, less prone to microbial contamination during transport and safer and have low logistical costs for easy transport.

When water is used as the aqueous substrate, after a mixture of the phytosterol and the emulsifier is dispersed in water, the resulting dispersion is evaporated, lyophilized or spray dried to produce an aqueous phytosterol powder. This powder is then used in an aqueous beverage to provide a beverage containing phytosterols.

The phytosterol and sucrose fatty acid ester are added to water by heating the resulting mixture before mixing, and then subjected to a stirring process and a high-pressure homogenization process in a predetermined order to obtain a transparent phytosterol dispersion. When the amount of phytosterol is 1%, the dispersion stability of the resulting solution cannot be ensured by the conventional emulsification process, causing the sedimentation of phytosterol. Conventional emulsification processes produce dispersions with light transmission as low as 0.16% at 700nm, whereas according to the method of the present invention, light transmission at 700nm is 80.0% or higher.

Regarding the mixing of the mixture of phytosterol and emulsifier with the aqueous beverage, the mixture of phytosterol and emulsifier may be added to the aqueous beverage as a hot liquid phase or as a solid phase that is cooled to room temperature. In the former case, the aqueous beverage is heated to 60-140 ℃, preferably to 70-90 ℃ to enhance its emulsification efficiency. The optimum temperature for the liquid phase mixture to be added directly to the beverage is 80 ℃. Specifically, in order to obtain small micelle particles, the heating temperature of the aqueous beverage is preferably set to be similar to the temperature of the mixture of the phytosterol and the emulsifier. After emulsification in water, the temperature was raised to 100 ℃ under pressure. For example, about 5 atmospheres of pressure are required to emulsify the phytosterol mixture in water at 140 ℃.

The mixture was stirred to form nanoparticles. In this regard, the stirring is carried out at 5,000-10,000rpm, preferably 6,500-7,500rpm, for about 10 minutes. The resulting micelles after the stirring process were tested, in which 90% or more of the particles had a size of 300nm or less. In contrast, the micelles obtained under the same conditions, except that the mixture of phytosterol and emulsifier was not heated, were measured to have a particle size in the range of tens to hundreds of microns. Therefore, the results of these comparative measurements indicate that the process of melting the phytosterol and emulsifier and mixing them is important for the formation of nanoparticles. In addition, as will be described below, high speed stirring or homogenization processes are also important to the formation of uniformly sized particles.

In the absence of other components, the phytosterol and the emulsifier can be uniformly contacted with each other and melted after being heated, so that the micelle particle size obtained after emulsification is hundreds of nanometers. Contrary to the conventional art, the present invention can produce nanoparticles suitable for use in beverages without using an organic solvent in which phytosterols are soluble.

After stirring, a homogenization process is required to grind the aggregated micelles into a powder. This homogenization process can be carried out by means of a high-pressure homogenizer, a colloid mill or a sonicator, with a high-pressure homogenizer being preferred. In accordance with the present invention, the micelles are homogenized in a homogenizer at a pressure of 2,000-. By this procedure, the resulting micelles were determined to have 95% or more of the micelle particles having a size of 300nm or less.

In accordance with the present invention, the desired plant sterol-containing beverage is obtained by hot melting the plant sterol and the emulsifier, mixing, stirring the mixture in water and treating under high pressure, and diluting the resulting dispersion with a fruit juice beverage, soda water, milk, soy milk, a beverage made from grains, or other popular beverages. In these beverages, the micelle particle size is small to nanometer, has a large surface area and particle curvature, is excellent in bioavailability, and does not affect the characteristic taste and flavor of the beverage.

In addition, the beverage prepared according to the present invention does not undergo stratification even when stored in a refrigerator, because the dispersion stability of the phytosterol micelles has been improved. More importantly, the phytosterol micelle keeps excellent dispersion stability at 90 ℃, so that the long-term stability of the product is ensured.

The invention will be better understood from the following examples, which are intended to illustrate and not to be construed as limiting the invention. In the following examples, particle size distributions were analyzed using a Mastersizer (malvern instrument ltd., UK).

Comparative example 1

To a1 liter vessel, 500 grams of water was added, heated to about 80 ℃, and then 5 grams of phytosterols (75% sitosterol, 10% campesterol, 15% stigmasterol sitostanol) and 4.25 grams of sucrose stearate (HLB11) were added to the hot water, followed by stirring the mixture at 6,800-. The resulting particle size was analyzed, and the results are shown in table 1 below.

TABLE 1

Particle size (μm)	Cumulative%
		0.985	0.07
1.89	12.41
		2.50	32.52
3.31	53.23
		4.38	66.82
5.27	77.28
		6.35	85.32
11.11	95.69
		21.32	98.96
78.56	100.00

Comparative example 2

The dispersion from comparative example 1 was treated with a high pressure homogenizer, model "Microfluidizer M110 EHI", such as that manufactured by Microfluidics, at a pressure of 7,000psi, through one pass. The resulting particle size was analyzed and the results are shown in table 2 below. The resulting dispersion has a light transmission of 0.16%, measured at 700 nm.

TABLE 2

Particle size (μm)	Cumulative%
		0.985	0.03
1.89	11.25
		2.50	30.43
3.31	54.47
		4.38	66.55
5.27	79.74
		6.35	88.45
11.11	96.21
		21.32	99.46
94.65	100.00

Comparative example 3

A1 liter container was charged with 40 grams of phytosterols (75% sitosterol, 10% campesterol, 15% stigmasterol sitostanol), 36 grams of vegetable oil and 4 grams of polysolvate 60(HLB 14.9) and melted to clarity with stirring at 130 ℃. sup.140 ℃. After melting was complete, 320 grams of water maintained at about 80 ℃ was added to the clear melt, which was then stirred at about 10,000rpm for about 10 minutes. It was observed that the resulting dispersion of phytosterols had particles of phytosterols floating on it, while a large number of particles of phytosterols appeared to stick to the walls of the container. The particle size distribution (volume) of the dispersion was analyzed, and as a result, 91% or more of the total number thereof had a particle size of 1 μm or more, and about 81% or more of the total number thereof was 100 μm or more. In addition, sedimentation and demixing were observed within 1 hour at room temperature, confirming that this phytosterol dispersion could not be used in beverages.

Comparative example 4

A1 liter container was charged with 15 grams of phytosterols (75% sitosterol, 10% campesterol, 15% stigmasterol sitostanol) and 30 grams of monoglycerol citrate (HLB 8.0) and melted to clarity with stirring at 130-. Water, maintained at about 80 deg.C, was then added to this clear melt, followed by stirring at about 6,800rpm for about 10 minutes. The resulting solution is treated through a channel at a pressure of 10,000psi using a high pressure homogenizer, model "Microfluidizer M110 EHI", such as manufactured by Microfluidics. In the phytosterols thus obtained, a large number of particles of phytosterols were clearly seen sticking to the walls of the container, since the phytosterol dispersion was not bound to the protein but to the water. The viscosity of the dispersion was as high as 58cps at a phytosterol concentration of 1%. The dispersion was analyzed for particle size distribution (volume) and the dispersion was determined to have a particle size of 1 μm or more when the cumulative percentage was 68% or more; when the cumulative percentage is 21% or more, the particle size is 10 μm or more, and such large particles are sufficient to give a mouth feel like bristles. In addition, its dispersion stability is poor, making it difficult to use the dispersion for preparing beverages.

Comparative example 5

A1 liter vessel was charged with 15 grams of phytosterols (75% sitosterol, 10% campesterol, 15% stigmasterol sitostanol) and 15 grams of Sodium Stearoyl Lactylate (SSL) and melted to clarity with stirring at 130-. 300 grams of water maintained at about 80 ℃ was then added to this clear melt, followed by stirring at about 6,800rpm for about 10 minutes. The resulting solution is treated through a channel at a pressure of 10,000psi using a high pressure homogenizer, model "Microfluidizer M110 EHI", such as manufactured by Microfluidics. Observing the phytosterol dispersion that settles within one day and completes solidification within three days, these results make this dispersion difficult to use in preparing beverages. The dispersion was analyzed for particle size distribution (volume) and the dispersion was determined to have a particle size of 1 μm or more when the cumulative percentage was 57% or more; when the cumulative percentage is 17% or more, the particle size is 10 μm or more. Such poor dispersion stability makes this dispersion difficult to use in preparing beverages.

Example 1

A vessel was filled with phytosterols (75% sitosterol, 10% campesterol, 15% stigmasterol sitostanol) and sucrose stearate (HLB11) and/or sorbitan lauryl ester (HLB 8.6) and melted at 130 ℃. 140 ℃. After completion of the melting, the solution was stirred for another 1 minute, added to water maintained at about 80 ℃ and then stirred at 6,800-. The resulting solution is treated through a channel at a pressure of 7,000psi using a high pressure homogenizer, model "Microfluidizer M110 EHI", such as manufactured by Microfluidics. Table 3 lists the amounts of phytosterol, sucrose stearate and sorbitan lauryl ester used to specify whether or not the high pressure homogenization process was performed.

TABLE 3

Numbering	Plant sterol	Sucrose stearate	Sorbitan lauryl ester	Water (W)	High pressure homogenisation
						1	5g	4.25g	-	500g	Whether or not
2	5g	4.25g	-	500g	Is that
						3	5g	3.036g	1.214g	500g	Whether or not
4	5g	3.036g	1.214g	500g	Is that
						5	25g	2.5g	1.75g	500g	Whether or not
6	25g	2.5g	1.75g	500g	Is that
						7	25g	2.126g	2.124g	500g	Whether or not
8	25g	2.126g	2.124g	500g	Is that

The particle size distribution analysis was performed on the solution of number 1 in Table 3, and the results are shown in Table 4 below.

TABLE 4

Particle size (μm)	Cumulative%
		0.096	20.35
0.127	52.19
		0.153	68.49
0.184	75.29
		0.222	85.33
0.294	91.52
		0.985	99.21
5.27	100.0

For reference purposes, the results of the analysis of particles numbered 3, 5, 7 in table 3 are similar to the results of number 1 shown in table 4.

In the case of number 2 in table 3, the analysis results of the particles are shown in table 5 below.

TABLE 5

Particle size (μm)	Cumulative%
		0.096	13.67
0.127	49.40
		0.153	69.39
0.184	77.61
		0.222	89.07
0.294	95.22
		0.985	99.89
2.08	100.0

For reference purposes, the results of the analyses of numbers 4, 6,8 in Table 3 are similar to the results of number 2 shown in Table 5.

The phytosterol dispersions prepared in example 1 (nos. 2, 4, 6 and 8) had a light transmittance measured at 700nm ranging from 80.0% to 80.5%.

Example 2

A vessel was filled with phytosterols (75% sitosterol, 10% campesterol, 15% stigmasterol sitostanol) and sucrose stearate (HLB11) and/or sorbitan lauryl ester (HLB 8.6) and melted at 130-140 deg.C with stirring. After completion of the melting, the solution was stirred for another 1 minute, added to water (80-90 ℃ C.) containing 1 g of sucrose stearate, and then stirred at 6,800-. The resulting solution was treated with a high pressure homogenizer (Microfluidizer M110EHI, Microfluidics) at a pressure of 7,000psi through one channel. Table 6 lists the amounts of phytosterol, sucrose stearate and sorbitan lauryl ester used to specify whether or not the high pressure homogenization process was performed.

TABLE 6

Numbering	Plant sterol	Sucrose stearate	Sorbitan lauryl ester	Water (W)	High pressure homogenisation
						9	5g	4.25g	-	500g	Whether or not
10	5g	4.25g	-	500g	Is that
						11	5g	3.036g	1.214g	500g	Whether or not
12	5g	3.036g	1.214g	500g	Is that
						13	25g	2.5g	1.75g	500g	Whether or not
14	25g	2.5g	1.75g	500g	Is that
						15	25g	2.126g	2.124g	500g	Whether or not
16	25g	2.126g	2.124g	500g	Is that

The particle size analysis was performed on the solution of number 9 in Table 6, and the results are shown in Table 7 below.

TABLE 7

Particle size (μm)	Cumulative%
		0.096	19.21
0.127	52.30
		0.153	68.72
0.184	76.41
		0.222	85.95
0.294	92.05
		0.985	99.35
4.80	100.0

For reference purposes, the results of the analysis of particles No. 11, 13 and 15 in table 6 are similar to the results of number 9 shown in table 7.

In the case of number 10 in table 6, the particle analysis results are shown in table 8 below.

TABLE 8

Particle size (μm)	Cumulative%
		0.096	14.50
0.127	48.24
		0.153	70.68
0.184	77.92
		0.222	90.61
0.294	96.74
		0.985	99.85
1.89	100.0

For reference purposes, the results of the analysis of particles No. 12, 14 and 16 in table 6 are similar to the results of number 10 shown in table 8.

The light transmittance of the phytosterol dispersions prepared in example 2 (nos. 10, 12, 14 and 16) was measured at 700nm and ranged from 80.5% to 82.5%.

Example 3

A container was charged with 5 grams of phytosterols (75% sitosterol, 10% campesterol, 15% stigmasterol sitostanol) and 4.25 grams of polyglycerol monostearate (HLB 12) and melted under stirring at 130-. After the melting was completed, the melt was stirred for another 1 minute and added to 490.75 grams of water heated to 80 ℃ followed by stirring at 6,800-. The resulting solution was treated with a high pressure homogenizer (Microfluidizer M110EHI, Microfluidics) at a pressure of 7,000psi through one channel.

The results of the particle size analysis before high pressure homogenization are the same as in table 4, within the experimental error allowed.

After high pressure homogenization, the results of the particle size analysis are the same as in table 5, within the allowable experimental error.

After high pressure homogenization, the light transmittance of the phytosterol dispersion was measured at 700nm and ranged from 80.0% to 80.5%.

Example 4

A container was charged with 5 g of phytosterol (75% sitosterol, 10% campesterol, 15% stigmasterol sitostanol) and 3.25 g of polyglycerol monostearate (HLB 12) and melted under stirring at 130-. After completion of the melting, the melt was stirred for another 1 minute and added to 491.25 g of water (80-90 ℃ C.) containing 1 g of polyglycerol monostearate, followed by stirring at 6,800-. The resulting solution was treated with a high pressure homogenizer (Microfluidizer M110EHI, Microfluidics) at a pressure of 7,000psi through one channel.

The results of the particle size analysis before high pressure homogenization are the same as in table 7, within the experimental error allowed.

After high pressure homogenization, the results of the particle size analysis are the same as in table 8, within the experimental error allowed.

After high pressure homogenization, the light transmittance of the phytosterol dispersion was measured at 700nm and ranged from 80.0% to 82.5%.

Example 5

The dispersions prepared in examples 1 to 4 were spray dried to obtain aqueous phytosterol powder.

Example 6

100 grams of the dispersion prepared in each of examples 1-4 was diluted with 400 grams of an aqueous beverage selected from the group consisting of water, juice beverages, soda pop, milk, soy milk, beverages made from grains, and other popular beverages. 9.25 grams of the powder prepared in example 5 was added to 490.75 grams of an aqueous beverage selected from the group consisting of water, juice beverages, soda pop, milk, soy milk, beverages made from grains, and other popular beverages. The obtained diluted beverages were taken by 50 professional technical subjects (20 men aged 30 and 40, 20 women aged 30 and 40, and 10 unbounded women) to test their performance by feeling. Fruit juice (orange juice) was used as a comparative beverage. The results are shown in tables 9 and 10 below.

TABLE 9

Comprehensive testing

	Is superior to the traditional beverage	As with conventional beverages	Is inferior to the traditional beverage
				Number of people	9	40	1
Percentage of	18％	80％	2％

Watch 10

Taste testing

Feeling of	Great agreement	Identity of a person	Neutral (neutral)	All differences are recognized	Very different recognition
						Acid(s)	2	6	30	11	1
Sweet taste	1	3	43	2	1
						Bitter taste	0	1	2	5	42
Has a bristle feeling	0	0	0	3	47

When other beverages were tested, the results were similar to tables 9 and 10.

Example 7

The beverage prepared in example 6 was stored in a refrigerator (4 ℃) for 1 year or more. Separately, the same beverages were treated at 90 ℃ for 4 hours. Not only was the beverage observed to remain in a normal dispersed state, but no other abnormalities were found.

Example 8

A vessel was charged with 5 g of phytosterol and 4.25 g of sucrose stearate (HLB11) and melted at 130 ℃ and 140 ℃ with stirring. After the melting is completed, stirring is carried out for 1-2 minutes. To the resulting solution was added 490.75 grams of an aqueous beverage selected from the group consisting of water, juice beverages, soda pop, milk, soy milk, beverages made from grains, and other popular beverages. 50 professional technical subjects (20 men aged 30 and 40, 20 women aged 30 and 40, and 10 unbounded women) took this diluted drink to test their performance by feel. Fruit juice (orange juice) was used as a comparative beverage. The results are shown in tables 11 and 12 below.

TABLE 11

Comprehensive testing

	Is superior to the traditional beverage	As with conventional beverages	Is inferior to the traditional beverage
				Number of people	6	42	2
Percentage of	12％	84％	4％

TABLE 12

Taste testing

Percentage of	Great agreement	Identity of a person	Neutral (neutral)	All differences are recognized	Very different recognition
						Acid(s)	3	5	31	10	1
Sweet taste	1	3	43	2	1
						Bitter taste	0	1	3	4	42
Has a bristle feeling	0	0	1	4	45

When other beverages were tested, the results were similar to tables 11 and 12.

Example 9

A vessel was charged with 5 g of phytosterol (melting point: 143 ℃ C.) and 4.25 g of sucrose stearate (HLB11), and melted under stirring at 130 ℃ and 140 ℃. After complete melting, stirring for another 1-2 minutes. The resulting homogeneous melt was cooled to room temperature to give a solid, which was then ground to a powder.

Example 10

The melting point of the powder prepared in example 9 was 125 ℃ as determined by a melting point tester (Electrothermal 9200).

Example 11

9.25 g of the powder prepared in example 5 or 9 was added to 90.75 g of water heated to 80-90 ℃ and stirred at 6,800-7,000rpm for 10 minutes.

Example 12

The solution prepared in example 11 was treated with a high pressure homogenizer (microfluidizer M110EHI, Microfluidics) at a pressure of 7,000psi through one channel.

Example 13

100 grams of the dispersion prepared in example 11 or 12 was diluted with 400 grams of an aqueous beverage selected from water, juice beverages, soda pop, milk, soy milk, beverages made from grains, and other popular beverages.

Example 14

9.25 grams of the dispersion prepared in example 5 or 9 was added to 490.75 grams of an aqueous beverage selected from the group consisting of water, juice beverages, soda, milk, soy milk, beverages made from grains, and other popular beverages, and stirred at 6,800 and 7,000rpm for about 10 minutes.

Example 15

The beverage prepared in example 14 was treated with a high pressure homogenizer (microfluidizer m110EHI, Microfluidics) at a pressure of 7,000psi through one channel.

Example 16

The beverages prepared in examples 14 and 15 were stored at room temperature for 1 year or more, stored in a refrigerator (4 ℃) for 1 year or more, and left at 90 ℃ for 4 hours while monitoring their physical states. Not only was the beverage observed to remain normally dispersed, but no other abnormalities were found.

Example 17

Clinical trial

A beverage obtained by mixing the dispersion of number 2 in example 1 with coffee, milk or green tea was used for clinical trials according to the method of example 4.

1. Clinical subjects: 45 people suffer from mild hyperlipidemia.

Watch 13

Condition of the subject before clinical trial

Surname pin (Man: woman)	15∶30
		Age (year of old)	56.53±10.28
Body Mass Index (BMI) (kg/m))	25.14±2.51
		Ideal weight Percentage (PIBW)	119.20±12.60
Total serum cholesterol (mg/L)	246.12±27.61
		Neutral lipids (mg/L)	145.81±57.43
LDL-Cholesterol (mg/L)	161.90±19.35
		HDL-cholesterol (mg/L)	52.89±12.54

2. Dietary control of clinical subjects: no special diet is provided; providing a low cholesterol diet to a patient suffering from mild hyperlipidemia.

TABLE 14

Dietary Condition of clinical Subjects

Total heat (Cal)	1705.9±330.49
		Protein (g)	64.26±18.53
Total lipids (g)	40.12±17.26
		Saturated lipids (g)	11.12±6.13
Cholesterol (mg)	135.82±80.13

3. Prescription

Green tea or coffee containing 1.6 grams of phytosterol per bottle was used for clinical trials.

3-1 test group 1

Step 1: from week 0 to week 4,1 bottle of beverage was taken per day.

Step 2: from week 5 to 8, 2 bottles of beverage per day.

3-2 test group 2

Step 1: from week 0 to 4,1 bottle of placebo was taken per day.

Step 2: from week 5 to 8, 1 bottle of beverage per day.

And step 3: from week 9 to week 12, 2 bottles of beverage per day.

4. Results

45 persons (male: female: 15: 30, average age: 56) were performed exactly according to the instructions and conditions of the clinical trial. In the initial phase, clinical subjects were measured to ingest 11.12 grams of saturated fat and 135.8 milligrams of cholesterol on a daily average diet. In the placebo-ingested test group and the 8-week-ingested phytosterol test group, the treatment effect was observed as the total serum cholesterol (p ═ 0.039) and LDL-cholesterol (p ═ 0.036) contents. In these groups, the periodic and residual effects of serum lipid content (total cholesterol, neutral lipids, HDL-cholesterol, LDL-cholesterol) were not statistically significant. After 8 weeks of intake of phytosterols, the patients were tested for a 4.38% decrease in total cholesterol (p ═ 0.039) and an 8.28% decrease in LDL-cholesterol (p ═ 0.036). In clinical subjects, the total cholesterol content of 33 persons decreased by an average of 9.2%, while the LDL-cholesterol content of 31 persons decreased by an average of 14.1%.

5. Conclusion

Hyperlipidemia patients can lower the total serum cholesterol and LDL-cholesterol levels after taking the plant sterol-containing beverage of the present invention for 8 weeks. This reduction can be obtained in patients who have a low dietary cholesterol intake.

Industrial applicability

As described above, according to the present invention, phytosterol nanoparticles are formed by thermally melting phytosterol with at least one emulsifier selected from the group consisting of sucrose stearate fatty acid ester, sorbitan fatty acid ester and polyglycerin fatty acid ester, and dispersing the molten mixture in an aqueous substrate. The beverage containing phytosterol can inhibit intestinal absorption and bile cholesterol absorption, thereby reducing serum cholesterol content. According to the present invention, phytosterol micelles with a particle size of hundreds of nanometers or less can be prepared without an organic solvent, and thus can be used in aqueous beverages. Since the phytosterol micelles of the present invention are formed of particles having a size of several hundred nanometers or less, the micelles have a high bioavailability. In addition, the improvement of the dispersion stability of the phytosterol micelle can prolong the life cycle of the beverage and ensure the long-term stability of the product. Further, such micelles can be used in almost all beverages regardless of the beverage base and pH. Since the micelles do not affect the characteristic taste and flavor of the beverage and the mouthfeel is good, the micelles can be sufficiently used in various aqueous beverages.

Claims (38)

1. A method of dispersing phytosterols comprising the steps of:

hot-melting a mixture of phytosterols and at least one emulsifier selected from sucrose fatty acid esters, sorbitan fatty acid esters and polyglycerin fatty acid esters at 60-200 deg.C;

mixing the molten mixture with an aqueous beverage or an aqueous beverage containing an emulsifier; and

stirring the mixture at a speed of 5,000 to 10,000rpm to produce a dispersion of phytosterols in the beverage,

whereby the phytosterol can be dispersed into 90% or more of particles having a particle size of 300 nm.

2. The method of claim 1, further comprising the step of:

homogenizing the mixture at a pressure of 2,000-25,000psi after the step of stirring the mixture at a speed of 5,000 to 10,000 rpm.

3. The method of claim 1, characterized in that the method comprises the steps of:

cooling the melt to a solid between the step of hot melting and the step of stirring the mixture, grinding the solid to a powder and mixing the powder with an aqueous beverage or an aqueous beverage containing an emulsifier.

4. The method of claim 3, further comprising the steps of:

homogenizing the mixture at a pressure of 2,000 to 25,000psi after the step of agitating the mixture at a speed of 5,000 to 10,000 rpm.

5. A method as claimed in any one of claims 1 to 4, wherein 95.0% or more of the particles have a size of 300nm or less.

6. A process as set forth in claim 1 or 2 further comprising the step of drying the dispersion to produce an aqueous phytosterol powder.

7. A process as claimed in claim 6, wherein the drying step comprises evaporation or freeze drying or spray drying.

8. The method as set forth in any one of claims 1 to 4, wherein the phytosterol is at least one selected from the group consisting of sitosterol, campesterol, stigmasterol, sitostanol and campestanol.

9. A method as claimed in claim 8 wherein the phytosterol is sitosterol.

10. The method as set forth in any one of claims 1 to 4, wherein the emulsifier is sucrose fatty acid ester.

11. The method as set forth in claim 10, wherein the sucrose fatty acid ester has a hydrophilic lipophilic balance value of 7 or more.

12. The method as set forth in claim 11, wherein the sucrose fatty acid ester has a hydrophilic lipophilic balance value of 10 to 16.

13. The method as set forth in any one of claims 1 to 4, wherein the stirring step is carried out at a speed of 6,500 and 7,500 rpm.

14. The method as claimed in any one of claims 1 to 4, wherein the aqueous beverage is water, a fruit juice beverage, a carbonated drink, milk, soy milk, a beverage made of grains, a beverage liked by other people or an alcoholic beverage.

15. The method as claimed in claim 14, wherein the aqueous beverage is water.

16. The method as set forth in any one of claims 1 to 4, wherein the melting step is carried out at 120-150 ℃.

17. A process as claimed in any one of claims 1 to 4 wherein the phytosterols are mixed with the total emulsifier in a weight ratio of 1: 0.01-10.

18. A method as claimed in claim 17, wherein the phytosterols are mixed with the total emulsifier in a weight ratio of 1: 0.2-2.0.

19. The method as set forth in any one of claims 1 to 4, wherein the emulsifier mixed with the aqueous beverage is used in an amount of 80% by weight or less based on the weight of the emulsifier mixed with the phytosterol.

20. The method as set forth in any one of claims 1 to 4, wherein the phytosterol is mixed with the aqueous beverage in a weight ratio of 1: 10-10,000 (w/w).

21. A method as claimed in claim 20 wherein the phytosterol is mixed with the aqueous beverage at a weight ratio of 1: 10-100 (w/w).

22. The method as set forth in any one of claims 1 to 4, wherein the composition of the mixture of the plant sterol and the emulsifier with the aqueous beverage is maintained at 60 to 140 ℃.

23. The method as claimed in claim 22, wherein the composition is maintained at 70-90 ℃.

24. A method as set forth in any one of claims 1-4 wherein the dispersion has a light transmittance of 80.0% or greater at 700 nm.

25. A method as claimed in claim 2 or 4, wherein the homogenisation step is carried out using a high pressure homogeniser, a colloid mill or a sonicator.

26. A method as claimed in claim 25, wherein the homogenising step is carried out using a high pressure homogeniser.

27. The method as set forth in claim 2 or 4 wherein the high pressure homogenization is operated at 7,000-10,000 psi.

28. A phytosterol dispersed beverage prepared by the method of any one of claims 1-4.

29. A phytosterol dispersed beverage as described in claim 28 wherein the beverage is a beverage made from water, juice beverages, carbonated drinks, cow's milk, soy milk, grains, other popular beverages, or alcoholic beverages.

30. An additive suitable for beverages, characterized in that it is prepared by heat-melting phytosterol and at least one emulsifier selected from the group consisting of sucrose fatty acid esters, sorbitan fatty acid esters and polyglycerol fatty acid esters at 60-200 ℃ and forming particles having a size of 300nm or less when dispersed in an aqueous beverage by stirring at 5,000-10,000rpm and then homogenizing at a pressure of 2,000-25,000 psi.

31. An additive as claimed in claim 30, wherein 90% or more of the particles have a size of 300nm or less.

32. An additive as claimed in claim 31, wherein 95% or more of the particles have a size of 300nm or less.

33. An additive as claimed in claim 30, wherein the additive is in the form of a powder.

34. The additive as set forth in claim 30, wherein the mixture is heated at 130-140 ℃.

35. The additive as set forth in claim 30, wherein the phytosterol is at least one selected from the group consisting of sitosterol, campesterol, stigmasterol, sitostanol and campestanol.

36. The additive as set forth in claim 30, wherein the emulsifier is sucrose fatty acid ester.

37. The additive as set forth in claim 36, wherein the sucrose fatty acid ester has a hydrophilic-lipophilic balance value of 7 or more.

38. The additive as set forth in claim 37, wherein the sucrose fatty acid ester has a hydrophilic-lipophilic balance value of 10 to 16.