Foodstuffs and pharmaceuticals

The present invention relates to a process for preparing foodstuffs or pharmaceuticals containing a definite amount of a-monolinolein and/or β-monolinolein (hereafter collectively referred to as monolinolein). The foodstuffs and pharmaceuticals in accordance with the present invention possess an excellent action of lowing choleste- level rol in blood or an excellent action of suppressing an level increase of cholesterol in blood.

level The action of lowering cholesterol in blood as used herein refers to an action of lowering cholesterol level in blood in the case where the cholesterol level is higher than normal cholesterol level.. Further the action level of suppressing an increase of cholesterol in blood as used level herein refers to an action of suppressing cholesterol in blood from constantly increasing even though foodstuffs containing large amounts of cholesterol are uptaken.

The term "foodstuff" as used in the present invention collectively refers to food edible to normal people including processed foodstuffs containing oils and fats (preferably foodstuffs containing at least 1.0 wt% of oils and fats therein) such as margarine shortening, mayonnaise, dressings, soup, chocolates, caramels, biscuits, cakes, donuts, breads, tofu or bean curd, rice cakes, etc., dairy products obtained by processing milk to be edible, such as cheese, butter, cream, condensed milk, powdered milk, fermented milk, ice cream, etc., beverages such as soybean milk, milk, juice, refreshing beverages, nutrient drink, carbonated drink, etc.

Based on findings in the past, it is a well known fact that polyunsaturated fatty acids (linoleic acid, etc.) contained in plant oils, plant sterols and the like possess an action of lowering cholssteroi or suppressing an increase of cholesterol in human and other animal bloods.

In recent years, reduction to practice has been attained as healthy foodstuffs in which substances of this type are supplemented and reinforced in foodstuffs or beverages for purposes of improving nutrition. Further in the field of pharmaceuticals, attention has been brought to . physiological and pharmacological effects of these substances and in light of extensive investigations and results, pharmaceuticals have been developed and provided for use.

The present inventors previously filed Japanese Patent Application OPI Nos. 35517/82, 36935/82, 36936/82, 39736/82, 42631/82, etc. 'However, effective ingredients contained in compositions called for in these applications and edible oils and fats are contained in very small contents and it has not been clarified which compound would be effective. That is, these applications are directed to processes which comprises diluting a germ oil with an organic solvent, then cooling the dilution to -30 to -90°C, separating solid fractions, and fractionating from the solid fractions not eluted with a solvent having a solubility parameter of less than 7.5 but eluted with a solvent having a solubility parameter of not less than 7.5, stating that the action of lowing level level in bloody cholesterol or suppressing an increase of cholesterol is found in the thus collected fraction. However, it has not been clarified what would be an effective substance(s) in this fraction. Further, the prior art processes involve drawbacks that the yield of an effective substance(s) varies depending upon conditions for fractional procedures, etc.

Accordingly, an object of the present invention is to survey which is an effective substance in fraction mixtures in the aforesaid applications and to prepare foodstuffs or pharmaceuticals containing a specific amount of the thus surveyed effective substance and prepare foodstuffs or pharmaceuticals exhibiting an action of lowering cholesterol or an action of suppressing an increase of cholesterol level in blood.

As a result of extensive investigations, paying attention to physiological effects of oil and fat components in the foregoing substances heretofore shown, the present inventors have found that monolinolein, a kind of monoglycerides, exhibits an action of lowering cholesterol level in blood in an amount of 1/100 that of substances conventionally employed and an action of suppressing an increase of cholesterol level in blood is equivalent thereto and, have accomplished the present invention.

That is, the present invention is directed to foodstuffs or pharmaceuticals which comprises incorporating α-monolinolein and/or B-monolinolein into foodstuffs or pharmaceuticals in an amount of, for example, 1.0 wt% to 98 wt% calculated as monolinolein.

The present invention' is further directed to foodstuffs or pharmaceuticals which comprises incorporating a-monolinolein and/or B-monolinolein into foodstuffs or pharmaceuticals in an amount of, for example, 1.0 wt% to 98 wt% calculated as monolinolein and then, granulating, softening, capsulating or tableting the same.

The present invention is also directed to foodstuffs having good taste and flavor which comprises incorporating a-monolinolein and/or 6-monolinolein in an amount of, for example, 1.0 wt% to 98 wt% calculated as monolinolein into things edible to normal people including processed foodstuffs containing oils and fats (preferably foodstuffs containing at least 1.0 wt% of oils and fats therein) such as margarine, shortening, mayonnaise, dressings, soup, chocolates, caramels, biscuits, cakes, donuts, breads, tofu or bean curd, rice cakes, etc., dairy products obtained by processing milk to be edible, such as cheese, butter, cream, condensed milk, powdered milk, fermented milk, ice cream, -etc., beverages such as soybean milk, milk, juice, refreshing beverages, nutrient drink, carbonated drink, etc. in an amount of for example, 1.0 wt% to 98 wt.%, preferably 1.0 wt% to 30 wt%, calculated as monolinolein.

For preparing monolinolein employed in the present invention, naturally occurring monoglycerides can be obtained by isolation; further known synthesis methods including direct esterification or ester exchange of glycerin using catalysts such as alkalis, p-toluenesulfonic acid, etc. or enzymes and, partial hydrolysis of triglycerides or diglycerides can also be utilized. These methods can also be used in combination.

The monolinolein content contained in the compositions and edible oils and fats called for in the previous applications (Japanese Patent Application OPI Nos. 35517/82, 36935/82, 36936/82, 39736/82 and 42631/82) by the present inventors is less than 1.0% and the effect is poorer than in the present invention. Further, the compositions are semi-solid substances containing 40 to 50% of plant sterols in addition to monolinolein. Accordingly, even though the compositions are incorporated into foodstuffs, foodstuffs having good taste and flavor are not obtained.

The term monolinolein as used herein refers to a-monolinolein and/or ß-monolinolein. In particular, B-monolinolein has an excellent effect.

The thus obtained a-monolinolein and/or B-monolinolein is incorporated in an amount of 1.0 wt% to 98 wt%, preferably 2 wt% to 5 wt%, based on the total amount. Materials in which a-monolinolein and/or ß-monolinaiein is incorporated may be any materials as long as they are normally edible to human beings. That is, materials include animal and plant proteinous materials such as casein, soybean proteins, egg proteins, etc. Further, vitamin E, vitamin F, lecithin or the like may also be employed in combination. It is also possible to use monolinolein as a foodstuff, by incorporating it into oils and fats such as animal and plant oils, et-c., edible materials such as carbohydrates, etc.

In the case of using in foodstuffs, it is particularly preferred that monolinolein be incorporated into and dissolved in oils and fats. As oils and fats, there are employed all of oils and fats including plant oils and fats such as soybean oil, safflower oil, corn oil, cotton seed oil, rape seed oil, rice oil, palm oil, olive oil, sunflower oil, etc., animal oils and fats such as lard, tallow, cream, etc., a mixture thereof, or those processed by hydrogenation, ester exchange, fractionation, etc. In the case of incorporating monolinolein into beverages having a small content of oils and fats, monolinolein may also be employed in combination with other emulsifiers such as glycerine fatty acid esters, sucrose-fatty acid esters, sorbitan fatty acid esters, lecithin, etc..

The thus obtained formulation may be utilized as an injection, a fluid, etc. In addition, the formulation may also be employed after forming the same into medical preparations such as soft agents, granules, capsules, tablets, etc. In case that the formulation is converted into medical preparations, binders, vehicles and other auxiliary substances may be appropriately employed to mix them. Then, the resulting mixture can be converted into medical preparations such as tablets, capsules, granules, pills, etc. by means of a direct powder pressing method, a dry granule pressing method, a wet granule pressing method, etc. Examples of binders include starch, sugars, gelatin, gum arabic, methyl cellulose, etc. and examples of vehicles include calsium phosphate, calcium sulfate, sugars, starch etc. Monolinolein has no obstacle in accelerating a breakdown of medically preparaed forms and improving absorption in vivo due to emulsified dispersion.

These preparations can be utilized not only as single drugs but also as pharmaceuticals in combination with other drugs. Further, the medical preparations can also be employed as pharmaceuticals prepared for purpose of intravenous or rectal administration. In the case of using as a fluid for intravenous administration, at least three nutrients of fats, hydrated carbons and amino acids are incorporated and further vitamins, inorganic materials and the like are also formulated therein in a well balanced formulation from a dietetic viewpoint. That is, yolk phospholipids or soybean phospholipids are employed as emulsifiers in such a fluid; a part of or all of which can be replaced by monolinolein. Furthermore, monolinolein can be employed as emulsifiers for a sugar fluid, an amino acid fluid, a fat emulsion, etc. When monolinolein is employed in emulsifying fats in water under pressure, finely divided, stable oil-in-water type can be obtained. Further, monolinolein can also be incorporated, as a nutrient for rectal administration, into a nutrient food supplied through a tube, an elemental diet, etc., whereby the nutrient food is purified and absorbed in the intestinal tract.

In the present invention, monolinolein acts to lowering cholesterol in blood or suppress an increase of cholesterol level bood by uptake of monolinolein of the present invention as pharmaceuticals independently or in combination with other things, against hypercholesteremia due to various diseases such as arteriosclerosis, hyperlipemia, diabetes, liver disturbance, fatty liver, or, against increased cholesterol in blood which occurs even in normal people, when foodstuffs having a high cholesterol level are uptaken.

Monolinolein in accordance with the present invention has neither taste nor smell other than monoglyceride flavor which would be an obstacle in uptake and, is a highly stable substance.

Hereafter, the present invention will be explained in more detail with reference to the examples below.

Example 1

1) Preparation of Monolinolein:

- Preparation from Corn Oil -

To 1 kg of purified corn oil, 10 kg of methyl ethyl ketone was added. After stirring the mixture, the mixture was cooled to -70°C for 3 hours using a dry ice-methanol coolant. Thereafter, solidified substances were removed by suction filtration and methyl ethyl ketone was removed from the remaining methyl ethyl ketone solution by distillation using an evaporator to -obtain a red brown liquid. To the liquid, a 10-fold amount by weight of methyl ethyl ketone was added. Again by the same procedure, solidified substances at low temperatures were removed. Methyl ethyl ketone was removed by distillation to obtain 25 g of a red brown, highly viscous liquid.

22 g of this substance was developed, in sequence, with solvents shown in Table 1 through a glass column having an inner diameter of 70 mm in which 1500 g of activated "Wako Gel C-200" was packed. After recovering respective fractions, the solvents were removed by distillation to obtain four column-fractionated substances as shown in Table 2.

The thus obtained Fraction IV was eluted with methanol : isooctane : water = 500 : 10 : 20 using a high performance liquid chromatography eqipped with Hitach Gel 3053 Column to isolate a fraction corresponding to a retention time of standard monolinolein. The thus isolated fraction was monoglycerides mainly composed of linoleic acid. Isolation was repeated to obtain about 110 mg of monolinolein (purity 95%) from 7.6 g of Fraction IV.

2) Animal Test:

Male 28 days old rats of the Sprague-Dawley strain were fed a stock diet for 2 days. Then they were weighed to the nearest gram and divided into 2 groups of 10 rats on the basis of weight so that the average starting weight (81g) and weight range (78 to 84 g) were similar for each group. Animals were maintained individually in suspended cages in a room with regulated temperature (23 + 1°C) and relative humidity (55 + 10%). The light period was 12hrs. (from 7:OOAM to 7:00PM). Food and Water were available ad libitum∵and replenished.daily throughout the 19-days experimental period-Before feeding the diets were stored at 7°C. The composition of experimental diet is shown in Table 3. Monolinolein was mixed with tallow and soybean oil (50 : 50) melted in a hot water bath. At the end of the 19-days feeding period, the rats were fasted for 16 hours. Then, blood was collected under ethereal anesthesia and serum was collected in a conventional manner. The total cholesterol in serum and β-lipoprotein in serum were measured using a kit, "Cholesterol B-Test Wako", manufactured by Wako Junyaku Kogyo Co., Ltd. and a kit, "β-Lipoprotein-Test Wako", manufactured by the same company, respectively. Performance, total cholesterol in serum and ß-lipoprotein in serum are shown in Table 4.

From this study , no influence was noted on average weight gain, average feed consumed and feed efficiency due to the addition of manoliholein. In total cholesterol levels in serum and β-lipoprotein levels in serum, a significant effect was noted by the addition of monolinolein.

Example 2

1) Preparation of Monolinolein:

MONOLINOLEIN .(Approx. 99%) manufactured by SIGMA CHEMICAL COMPANY in U.S.A. was used in this experiment.

2) Animal Test:

After feeding male 28 days old rats of the Sprague-Dawley strain with commercial diet for 5 days, the animals were divided into groups. Using the rats weighing 104.2 + 4.1 g in average, each group being 10 rats, the rats were fed with each of test feeds for 17 days. Water was available ad libitum but food was consumed by paired feeding'(feeding was conformed to the minimum feeding group).

Tne composition of expeimental diet shown in Table 3, only tallow was employed in an amount of 10% as an oil and fat, groups feeding feeds to which 1% of cholesterol and 0.3% of cholic acid were added and not added were supplemented and, monolinolein was replaced for sucrose having three levels of 0.5%, 1.0% and 2.0% in feeds and added to the feeds. Blood collection and meansurements of total cholesterol and β-lipoprotein in'serum were carried out in a manner similar to Example 1.

From this study , even though the levels of adding monolinolein were varied, the average weight gain and the feed efficiency were not affected at all: also in findings after autopsy, no particular abnormality other than liver hypertrophy due to cholesterol load was noted. Effect of monolinolein added was noted in an amount of 1.0% in the feeds, with respect to total cholesterol and β-lipoprotein in serum.

Example 3

1) Preparation of Monolinolein:

MONOLINOLEIN (Approx. 99%) manufactured by SIGMA CHEMICAL COMPANY in U.S.A. was used in this experiment as

in Example 2.

2) Animal Test:

After feeding Male 28 days old rats of the Sprague-Dawley train with commercial diet for 5 days, the animals were divided into groups. Using the rats weighing 87.4 + 4.4g in average, each group being 10 rats, the rats were fed with each of test diets for 17 days. Water was available ad libitum but food was consumed by paired feeding.

The composition of experiment diet was the same as in Example 2: monolinolein was added in an amount of 2.1%, β-lipoprotein and linoleic acid were added in an amount of 1%, respectively, singly or incombination, to the feed, in a manner similar to Example 1. Blood collection and measurements of total cholesterol and

β- Lipoprotein in serum were carried out in a manner similar to Example 1.

In this experiment, the following were purchased and employed:

• Linoleic acid: manufactured by Wako Junyaku Co, Ltd.

B-sitosterol: manufactured by Wako Junyaku Co., Ltd.

From this study, even though monolinolein, B-sitosterol and linoleic acid were added, the average weight gain and the feed efficiency were not affected at all; also in findings after autopsy, no particular abnormality other than liver hypertrophy due to cholesterol load was noted. With respect to total cholesterol and B-lipoprotein in serum, a significant effect was noted with the addition of monolinolein alone even in an amount of 2.1_%.

It was noted that this effect was 3 time greater than of a-sitosterol and more than 200 times that of linoleic acid.

Example 4

1) Preparation of Monolinolein:

MONOLINOLEIN (Approx. 99%) manufactured by SIGMA CHEMICAL COMPANY in U.S.A. was used in this experiment as in

Example 2.

2) Animal Test:

After feeding Male 28 days old rats of the Sprague--Dawley strain with commercial diet for 5 dats, the animals were divided into groups. Using the rats weighing 85.0 + 4.0g in average, each group being 10 rats,the rats were fed with each of test diets for 18 days. Food and water were available ad libitum. The composition of experimental diet is shown in Table 7. Blood collection and measurements of total cholesterol and β- lipoprotein in serum were carried out in a manner similar to Example 1.

From this study, no difference among the groups was noted either in feeding results of the animals or in findings after autopsy. A significant effect was observed in the total cholesterol level and β-lipoprotein level in serum when 1..8% of monolinolein was added to the feed.

Example 5

1) Preparation of Monolinolein:

MONOLINOLEIN (Approx. 99%) manufactured by SIGMA CHEMICAL COMPANY in U.S.A. was purchased and used in this experiment as in Example 2.

2) Animal Test: male 28 days old rats of the After feeding Sprague- Dawley strain with commercial diet for 5 days, the animals were divided into groups. Using the rats weighing 218.3 + 17.7 g in average, each group being 10 rats, the rats were ad libitum fed for 17 days with each of feeds having the same composition as shown in Table 3 except that tallow alone was added in an amount of 10% as the fat and oil, to which no monolinolein was added and, cholesterol and cholic acid were added in the same amounts as shown in Table 3. At this point of time, the total cholesterol in blood was measured with a part of the rats and, monolinolein was injected to the remaining rats from the tail vein once a day for three days under the condition that the aforesaid feed obtained by loading cholesterol to tallow was consecutively fed to examine any influence on the cholesterol in blood.

The injections prepared by adding monolinolein in concentrations of 0.2% and 0.5%, respectively, were given to the rats, using as a control a physiological saline solution containing 2 mg/ml of Tween 80.

From this results, it was clearly revealed that the administration of monolinolein by injection lowered the total cholesterol levels in serum.

Example 6

1) Preparation of Capsules. Tablets and Granules:

• (1) Preparation of Monolinolein:

  • MONOLINOLEIN (Approx. 99%) manufactured by SIGMA CHEMICAL COMPANY in U.S.A. was purchased and used in this experiment as in Example 2.

• (2) Preparation of Capsules:

  • 100 g of corn oil (manufactured by Ajinomoto Co., Ltd.) and 0.2 g of plant sterol (manufactured by _Tama Seikagaku Co., Ltd.) were heated for 30 minutes to melt them. After cooling, 5 g of linoleic acid (manufacatured by Wako Junyaku Industry Co., Ltd.) and 0.5 g of vitamin E (manufactured by Nisshin Kagaku Co., Ltd.) were mixed with the resulting melt to obtain Mixture A. Further, 1 g of monolinolein was mixed with Mixture A to obtain Mixture B. These two mixtures were encapsulated using 65 g of gelatin to obtain 500 mg/grain of Capsule A (no monolinolein was added) and Capsule B (monolinolein was added), respectively.

• (3) Preparation of Tablets:

  • To Mixtures A and B obtained in Preparation of Capsules, 500 g of corn starch (manufactured by Shionogi Pharmaceutical Co., Ltd.), 50 g of carboxymethyl cellulose (manufactured by Gotoku Yakuhin Co., Ltd.) and 40 g of hydroxypropyl cellulose having a low degree of substitution (manufactured by Shin-Etsu Kagaku Co., Ltd.) were added to mix. Further 5 ml of water was added and the mixture was kneaded. The kneaded system was grained and ground and then dried at 50°C over 6 hours. After adding 2 g of magnesium stearate (manufactured by Junsei Kagaku Co., Ltd.) to the dried system and mixing them, the mixture was tableted to obtain Table A (no monolinolein was added) and Table B (monolinolein was added).

• (4) Preparation of Granules:

  • To Mixtures A and B obtained in Preparation of Capsules, 35 g of corn starch, 65 g of carboxymethyl cellulose and 40 g of hydroxypropyl cellulose having a low degree of substitution were added to mix. Further 110 g of a 50% ethanol aqueous solution was added to the mixture to knead. After graining the thus kneaded system, it was blow-dried to remove ethanol. Then drying was performed at 30°C over 6 hours and sieved to obtain Granule A (no monolinolein was added) and Granule B (monolinolein was added), having a grain size of 1410 to 500 µm.

• ₂) Animal Test: strain

  • After feeding male 28 days old rats of the Sprague-Dawley with commercial diet for 5 days, the animals were divided into groups. Using the rats weighing 93.8 + 3.4 g in average, each group being 10 rats, the rats were fed with each of test feeds for 16 days. Water was available ad libitum but food was consumed by paired feeding.

The composition of experimental diets was the same as in Table 3 except that tallow was used in an amount of 10% as the oil and fat; for the sample addition groups, cholesterol and cholic acid were added in amounts of 1% and 0.3%, respectively. The capsules, tablets and granules were mixed with the feed after cutting the capsules with a surgical knife to expose the content and, grinding the tablets and the granules into powders in a motar.

Blood collection and measurement of the total cholesterol in serum were carried out in a manner similar to

Example 1.

From this study, even though 5% of the capsules, tablets and granules prepared by adding B-s.itosterol and linoleic acid and further adding monolinolein to the foregoing two were added, the average weight gain and the feed efficiency were not affected at all; also in findings after autopsy, no particular abnormality other than liver hypertrophy due to cholesterol load diet was noted. An effect of the addition in each of the experimental on the total cholesterol in serum was remarkable with any of the feeds to which monolinolein was added.

Example 7

After feeding male 28 days old rats of the Sprague-Dawley strain with commericial diet for 2 days, the animals were into 4, divided groups. Using the rats weighting 94.4 + 3.52 g, one group being 10 rats, the rats were fed with each of test feeds for 18 days. Food and water were available ad libitum. The composition of experimetal diet was the same as that shown in Table 3. Monolinolein and the fourth component in Example 1, Table 3. Monolinolein and the fourth component in Example 1, Table 2 were mixed with tallow and soybean oil (50 : 50) melted in a hot bath. Edible oil and fat to which monolinolein was added showed good taste and flavor however, taste and flovor of an oil and fat to which the fourth component was added was not good.

Blood collection and the measurements of the total cholesterol and ß-lipoprotein in serum were carried out in a manner similar to Example 1.

The results are shown in Table 11.

Example 8

After feeding male 28 days old rats of the Sprague-Dawley strain with commercial diet for 2 days, the animals were grouped. Using the rats weighing 92.3 + 3.2 g, one group being 10 rats, the rats were fed with each of test feeds for 19 days. Food and water were available ad libitum The composition of experimental diet was the same as that shown in Table 3.

To 54 g of yolk, 60 ml of vinegar and 6 g of sodium chloride were added and the mixture was agitated with a whipper to form a homogeneous aqueous phase. The aqueous phase was divided into three portions, to a first portion of which 100 g of safflower oil was added, to a second portion 100 g of safflower oil having dissolved therein the fourth component of Example 1, Table 2 in an amount of 2.0% and further to a third portion 100 g of safflower having dissolved therein 1.0% of monolinolein was added to form O/W type emulsions. Using the emulsions, three kinds of mayonnaise were prepared. Each of them was mixed with the feed in an amount of 10 wt%, which was provided for test.

Blood collection and the measurements of the total cholesterol and ß-lipoprotein in serum were carried out in a manner similar to Example 1.

Example 9

After adding monolinolein to purified soybean oil and edible, hardened soybean oil (melting point, 34°C) having a composition shown in Table 13 and thoroughly mixing the resulting mixture, monostearin, lecithin, carotene and vitamin A were added to the mixture to dissolve them. To the resulting solution an aqueous solution of salt, fragments and sodium dehydroacetate was added while stirring to emulsify in W/O type. The resulting emulsion was quenched and kneaded to prepare margarin containing monolinolein therein.

The product has the quite the same appearance as ordinary margarine; taste, flavor, etc. were not inferior to ordinary margarine.

This margarin had the same suppressing effect on blood cholesterol level as that in the mayonnaise obtained in Example 8.

Example 10

1) Preparation of Dressing:

To purified corn oil monolinolein was added to dissolve it. To the solution, vinegar, yolk and spices were added and a paste of corn starch, salt and water was further added thereto. After emulsifying the mixture, the emulsion was passed through a colloidal mill to prepare a dressing.

Proportions for formulation are shown in Table 14.

This dressing had good flavor and test, and the same suppressing effect on blood cholesterol level as that in the mayonnaise obtained in Example 8.

Example 11

₁) Preparation of Monolinolein-Containing Biscuit:

Raw materials shown in Table 5 were mixed to prepare a base for biscuit.

After molding, the mold was put in an oven followed by baking at 220°C for 15 minutes.

This biscuit had the same suppressing effect on blood cholesterol level as that in the mayonnaise obtained in Example 8.

Example 12'

1) Preparation of Monolinolein-Containing Chocolate:

Raw materials shown in Table 16 were mixed and ground into fine grains. Thereafter, the grains were purified, tempered, casted and cooled to prepared chocolate.

The thus obtained chocolate had quite the same flavor, melting at the mount, taste, etc. as those of ordinary one.

This chocolate had the same suppressing effect on blood cholesterol level as that in the mayonnaise obtained in Example 8.

Example 13

After feeding male 28 days old rats of the Sprague-Dawley strain with commercial diet for 2 days, the animals were grouped. Using the rats weighing 104.3 ± 4.1 g, one group being 10 rats, the rats were fed with each of test feeds for 18 days. Food and Water were available ad libitum. The composition of experimental diet was the same as that shown in Table 3.

Fats were separated from milk using a cream separator. At the working step out of steps of preparing butter comprising steps of neutrallization of cream, sterilization, cooling, holding, charning, removal of butter milk, washing of butter, salt-addition, working, casting and wrapping, 20 g of the fourth component of Example 1, Table 2 and 10 g of monolinolein were added, respectively, per 1 kg of butter and subsequent steps were followed in a conventional manner to obtain butter. Three kinds of butter together with commercially available were added to the feed in an amount of 10 g each, which were provided for test.

Blood collection and the measurements of the total cholesterol and ß-lipoprotein in serum were carried out in a manner similar to Example 1.

The functional evaluation was conducted by five expert pannelers using a 5 point evaluation. The results are shown in Table 17..

.Example 14

In a process for preparing cheese which comprises heat-treatment of a mixture comprising proteins, fats and other milky components separated from a mixture of milk, cream, defatted milk, butter milk and whey using a rennet, casting, pressing, salt-addition and ripening, 15 g of monolinolein was added to the mixture obtained after heat-treatment per 1 kg followed by subsequent steps in a conventional manner. Thus cheese was prepared.

This cheese had the same suppressing effect on blood cholesterol level as that in the butter obtained in Example 13.

Example 15

After concentrating fresh, clean, defatted milk to a 2/3 volume that of the original volume, the defatted milk was heated at 90° C for 15 minutes. After cooling to 45°C, a starter for yogurt (having a ratio of B ulgariens and Str. thermophilus of 1:1) was added in an amount of 2% and the mixture was fermented at 45° C for 4 hours in a container which had been previously sterilized and cooled. After adding 1.2% of monolinolein thereto, the mixture was cooled at 5°C for 8 hours to obtain yogurt.

This yogurt had the same suppressing effect on blood cholesterol level as that in the butter obtained in Example 13.

Example 16

Ice cream was prepared by adding sugar and fragments to cream, adding gelatin thereto as a stabilizer, further adding egg and fruits, adding monolinolein to the resulting mixture and then freezing the mixture while blowing air thereto.

This ice cream had the same suppressing effect on blood cholesterol level as that in the butter obtained in Example 13.

While the invention has been described in detail end with reference to specific embodiments thereof, it will be apparent to one skilled in the art that various changes and modifications can be made therein without departing from the spirit and scope thereof.