JPH0913075A - Oil and fat for diminishing lipid in blood - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an oil and fat having a reducing effect on the concentration of lipids in blood by using triglycerides in which the fatty acid moieties comprise ones derived from at least one specific unsaturated fatty acid and the amount of the 2-position acid moieties derived from the specific acid is not larger than a specific value. SOLUTION: This oil and fat having a reducing effect on the concentration of lipids in blood comprises preferably at least 5wt.% triglycerides in which the fatty acid moieties comprise ones derived from at least one long-chain polybasic unsaturated fatty acid of the n-3 type (a) and the amount of the 2-position acid moieties derived from the acid (a) is less than 40mol% of all units derived from the acid (a). The acid (a) is preferably at least one selected from the group consisting of α-linolenic acid, arachidonic acid, eicosapentaenoic acid, docosapentaenoic acid, and decosahexaenoic acid.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to fats and oils having the action of reducing blood lipid levels.

[0002]

BACKGROUND ART Cholesterol, triglyceride, phospholipid and free fatty acid are known as items for evaluating the lipid concentration in serum, and hyperlipidemia is a condition in which the lipid content of these is increased. An epidemiological study reveals that there is a positive correlation between serum cholesterol levels and the risk of developing ischemic heart disease, and that decreasing serum cholesterol also decreases the risk of developing ischemic heart disease. (For example, Yu Mizushima et al., “Today's therapeutic drugs (199
3rd year edition) ", p. 361, Nankodo). In addition to hypertriglyceridemia leading to the development of fatty liver, pancreatitis, etc., it has been pointed out that it is a risk factor for ischemic heart disease. Therefore, clinically, hypercholesterolemia and hypertriglyceridemia are particularly serious problems among hyperlipidemia.

When hyperlipidemia develops, generally, a hyperlipidemic patient should be treated with a dietary regimen such as caloric intake restriction.
After observing changes in serum lipid levels for 3 months, anti-high doses of clofibrate, cholestyramine nicotinate, etc. were mainly used to eliminate risk factors leading to arteriosclerotic diseases including coronary artery disease. Lipidemia agents have been administered to reduce serum cholesterol and triglyceride levels.

On the other hand, eicosapentaenoic acid (all cis-
5,8,11,14,17-eicosapentaenoic acid, hereinafter abbreviated as EPA. The numbers after C _{20: 5} and C represent the total number of carbons: the number of double bonds, and the same shall apply hereinafter. ) And docosahexaenoic acid (all
cis-4,7,10,13,16,19-docosahexaenoic acid, hereinafter abbreviated as DHA. It has been clarified by animal experiments and clinical experiments that n-3 long-chain polyunsaturated fatty acids such as C _{22: 6} ) and food materials containing them have an action of reducing serum triglyceride level and cholesterol level. (Eg Robinson, DR et al., J. Lipid Res., 34, 1435, 1993). The mechanism of action for reducing the triglyceride level in serum is that the intake of fats and oils containing n-3 long-chain polyunsaturated fatty acids suppresses the triglyceride synthesizing ability in the liver, resulting in the release of triglyceride into the blood. It is presumed that this is due to the suppression of oil (Kenji Hara, Fats and Oils, Vol. 46, No. 4, p. 90, 1993). It is also presumed that the decrease in serum cholesterol level is due to the fact that the n-3 long-chain polyunsaturated fatty acid suppresses the cholesterol synthesizing ability in the liver. (Choi, YS
Et al., Lipids, 24, 45, 1989).

Therefore, for the purpose of preventing hyperlipidemia and improving the serum lipid concentration of hyperlipidemia patients, intentionally ingesting foods containing a large amount of fish containing EPA and DHA, and EPA and DH
Health foods and the like made of fish oils containing A and fish oil concentrates are commercially available. However, it was necessary to ingest or administer these in large amounts and over a long period of time. As the fish oil containing EPA and DHA, sardine oil, cod liver oil, herring oil, squid oil, tuna orbital oil, etc. are mainly used, and the chemical structures of these oils and fats are ester bonds to glycerides. 50 mol% or more of the total amount of the -3 series long-chain polyunsaturated fatty acids is the constituent fatty acid at the 2-position of the triglyceride, in other words, the n-3 series long-chain polyunsaturated fatty acids are at the 1st and 3rd positions. It has a triglyceride structure with more ester bonds at the 2-position.

On the other hand, EPA and DHA have the effect of reducing serum lipids as described above, but on the other hand, they have many double bonds in the molecule as compared with ordinary fatty acids which constitute edible vegetable oils and fats, and thus are easily oxidized, It is also known that excessive intake causes harmful effects on the living body. It is presumed that when the peroxidation reaction of lipids progresses in the living body, the membranes of the living body are damaged and cause lesions such as ischemic disease, arteriosclerosis, cataract, cancer, Alzheimer's disease, collagen disease and amyloidosis.

[0007]

SUMMARY OF THE INVENTION The present invention has been made in view of such a situation as described above, and an object of the present invention is to have no side effects on humans and animals and to obtain a conventional n-3 system. An object of the present invention is to provide an oil or fat that has an action of reducing blood lipid concentration and facilitating improvement of blood lipid by ingesting a smaller amount than a long-chain polyunsaturated fatty acid source.

[0008]

Means for Solving the Problems As a result of intensive studies aimed at achieving the above object, the present inventors have found that n-3 long-chain polyunsaturated fatty acids at the 1-position and / or 3-position of a glyceride structure. Oils and fats containing a large amount of n-type long-chain polyunsaturated fatty acids are used as a source of n-3 type long-chain polyunsaturated fatty acids. It has been found that the above-mentioned object is achieved because the effect of reducing the triglyceride value is significantly high. The present invention has been completed based on such findings.

That is, the gist of the present invention is to include an n-3 long-chain polyunsaturated fatty acid as a constituent fatty acid of glyceride,
Less than 40 mol% of the total amount of n-3 long-chain polyunsaturated fatty acids consists of triglycerides bonded to the 2-position of glycerides, or contains the triglycerides to reduce the blood lipid concentration. It is a fat and oil with action.

The triglyceride featured in the present invention is n
In a triglyceride composed of a fatty acid containing a -3 long-chain polyunsaturated fatty acid and glycerin, n-3
When the total amount of the system long-chain polyunsaturated fatty acids is 100 mol%, less than 40 mol% thereof and any fatty acid other than the n-3 system long-chain polyunsaturated fatty acids form an ester bond at the 2-position of the triglyceride. And at least 60 mol% of the n-3 long-chain polyunsaturated fatty acid and any fatty acid other than the n-3 long-chain polyunsaturated fatty acid randomly or at the 1st and 3rd positions of the triglyceride. It is non-randomly distributed and has an ester bond.

The n-3 long-chain polyunsaturated fatty acid is an n-3 having 18 or more carbon atoms and 3 or more double bonds.
System linear unsaturated fatty acid, specifically, α-linolenic acid (C _{18: 3} ), octadecatetraenoic acid (C _{18: 4} , 6,9,
12,15-octadecatetraenoicacid), arachidonic acid (C _{20: 4} ), EPA (C _{20: 5} ), docosapentaenoic acid (C _{22: 5} , 7,10,13,16,19-docosapentaenoic acid)
), DHA (C _{22: 6} ) and the like. In the present invention, among these, one or two or more kinds of mixed fatty acids selected from the group consisting of α-linolenic acid, arachidonic acid, EPA, docosapentaenoic acid and DHA are preferable, and further EPA and / or Or D
HA is more preferred.

As the fatty acid other than the n-3 long-chain polyunsaturated fatty acid, any of short-chain, medium-chain and long-chain fatty acids and saturated and unsaturated fatty acids can be used. A straight-chain, medium- or long-chain saturated or unsaturated fatty acid having 6 or more carbon atoms is preferable. Such fatty acids include caproic acid (C _{6: 0} ), caprylic acid (C _{8: 0} ), capric acid (C _{10: 0} ), lauric acid (C _{12: 0} ), myristic acid (C _{14: 0} ), palmitic acid. (C _{16: 0} ), palmitooleic acid (C _{16: 1} ), stearic acid (C _{18: 0} ), oleic acid (C _{18: 1} ), elaidic acid (C _{18: 1} ), linoleic acid (C _{18 : 2} ), α'-linolenic acid (C _{18: 3} , 5,8,11-octadecatrienoic acid), γ-linolenic acid (C _{18: 3} , 6,9,12-octadecatrienoic acid), Rheostearic acid (C _{18: 3} , 9,11,13-octadecatrienoic acid), arachidic acid (C _{20: 0} ), gadoleic acid (C
_{20: 1} ), behenic acid (C _{22: 0} ), erucic acid (C _{22: 1} ), brassic acid (C _{22: 1} ), and the like. These fatty acids may be used alone or may be used as mixed fatty acids in any proportion. Of these, myristic acid, palmitic acid, stearic acid, oleic acid, and linoleic acid are preferable.

To produce the triglyceride of the present invention composed of the n-3 long-chain polyunsaturated fatty acid and other fatty acids described above, a chemical synthesis method, a transesterification method, or an extraction method from a natural product is used. The technology such as the above may be used.
Examples of the chemical synthesis method include a desired amount and composition of a fatty acid, a fatty acid anhydride or a fatty acid halide (fatty acid chloride) and glycerin, an acidic substance (hydrochloric acid, sulfuric acid, paratoluenesulfonic acid, etc.), an alkaline substance (sodium hydroxide). Esterification of metals (zinc, tin, titanium, nickel, etc.), metal oxides (zinc oxide, alumina, ferrous oxide, etc.), metal halides (aluminum chloride, tin chloride, etc.), etc. 100 to 250 ° C. in a nitrogen gas stream in the presence or absence of a catalyst
It is preferable that the esterification reaction is carried out for 1 to 25 hours while heating to room temperature and removing the produced water.

The esterification product may be subjected to alkali deoxidation treatment, activated carbon, activated clay, alumina, silica gel, if necessary.
Free fatty acids, monoglycerides, diglycerides, which are subjected to adsorption / fractionation treatment using an ion exchange resin or the like, solvent separation treatment using a hydrophilic organic solvent such as methanol or ethanol and / or a lipophilic organic solvent such as n-hexane or xylene, Impurities such as coloring substances and odorous components are removed, and these treatments are appropriately combined to obtain triglyceride
The content of the n-3 long-chain polyunsaturated fatty acid residues bonded to the position is the total of the n-3 long-chain polyunsaturated fatty acid residues bonded to the 1st, 2nd and 3rd positions of the triglyceride. 40 of content
The triglyceride component may be fractionated or concentrated so as to be less than mol%. The triglyceride of the present invention is
For example, it is desirable to blow steam under heating and decompression to perform deodorizing treatment.

In order to obtain the triglyceride of the present invention by utilizing the transesterification method, for example, triglyceride (component a-1) of fatty acid containing a large amount of n-3 long-chain polyunsaturated fatty acid as a raw material and n- Fatty acid (component a-2) which does not substantially contain or contains a small amount of 3 type long chain polyunsaturated fatty acid, lower alcohol ester (methyl ester, component a-2)
Ethyl ester and the like. The same applies hereinafter. Or a component a-2 triglyceride in a desired ratio, or a fatty acid triglyceride (component b-1) substantially containing no or a small amount of n-3 long-chain polyunsaturated fatty acid and n-. Fatty acids containing a large amount of 3 long-chain polyunsaturated fatty acids (component b-
2) or a lower alcohol ester of component b-2 is mixed in a required amount, and an alkaline substance such as sodium hydroxide and potassium hydroxide as a catalyst, a metal alcoholate (metal alkoxide) such as sodium methylate, sodium ethylate and lithium butyrate. ), An anion exchange resin such as a basic anion exchange resin or an acidic cation exchange resin, or a lipase, it is convenient to carry out the transesterification reaction. When transesterification is carried out using a specific lipase as a catalyst, a new fatty acid group can be selectively introduced into the 1-position and 3-position of the triglyceride as described later, which is desirable as a method for producing the triglyceride of the present invention. .

The raw materials for the transesterification are vegetable oils such as linseed oil, sesame oil, perilla oil, sardine oil, cod liver oil, fish oil such as herring oil, squid oil, tuna orbital oil, whales, seals, etc. as component a-1. Pressed or extracted oil obtained from the origin of marine mammals such as fur seals, milk fat of the animals, chlorella, spirulina, donariella, etc.Also genus Nannochloropsis (for example, Nannochloropsis oculata), genus Trustchitrium (for example, Thraustochytrium aureum
), Crypthecodinium (eg Crypthecodinium
cohnii), isochrysis (eg Isochrysis galban
a) Oils and fats extracted from microalgae belonging to, etc., oils and fats derived from microorganisms of the genus Mortierella, etc., and n-3 long-chain polyunsaturated fatty acids or the above-mentioned various fatty acids containing this in any proportion. (Refer to paragraph 0012)
Triglycerides of mixed fatty acids with can be used. Component a
As -2, various fatty acids or their derivatives described in paragraph No. 0012 can be used.

As the component b-1, there are triglycerides obtained from plants, animals, microorganisms, microalgae, etc., soybean oil,
Rapeseed oil, cottonseed oil, corn oil, palm oil, palm oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, olive oil, peanut oil, cacao butter, Chinese tallow, sal butter, shea butter, beef tallow,
Lard, hydrogenated oils and fats thereof, fractionated oils and fats, and the aforementioned component a
-2 triglyceride, medium chain fatty acid triglyceride and the like can be exemplified, and as the component b-2, there is a fatty acid obtained by the hydrolysis treatment of the component a-1.

In the transesterification reaction, for example, the raw materials are mixed in a molar ratio of component a-1: component a-2 = 1: 0.1.
5, component b-1: component b-2 = 1: 2 to 10 are mixed, and when an alkali or a metal alcoholate is used as a catalyst, it is in an anhydrous state and is 0.5 to 0.5 at 80 to 120 ° C. Allow transesterification for 3 hours. Similarly, when an ion exchange resin is used, it is also in an anhydrous state, but it is preferable to circulate and contact the raw materials in a column system at room temperature to 40 ° C. When using lipase as a catalyst, the water content in the raw material is set to 1% by weight or less, and commercially available lipase powder or a known carrier such as celite, diatomaceous earth, activated carbon, porous glass, ion exchange resin, chitosan, high Immobilized lipase immobilized on molecular gel, cellulose powder or the like is added, and transesterification reaction is performed at 20 to 80 ° C. for 0.5 to 20 hours.

As the lipase, those originating from the following microorganisms or originating from animal organs can be used. That is, Aspergillus (eg Aspergillus niger), Mucor (eg Mucor miehei), Candida (eg Candida cyrindracea), Pseudomonas (eg Pseudomonas fragi), Alcaligenes (eg,
Alcaligenes s described in JP-B-58-36953
p.), Rhizopus delemar (eg Rhizopus delemar), Geotrichum (eg Geotrichum candidum), etc., and lipases of microbial origin and pancreatic lipase of pigs, cattle, etc. Of these, lipases and porcine pancreatic lipases originating from microorganisms of the genus Aspergillus, genus Mucor, genus Alcaligenes and genus Rhizopus specifically act on the 1- and 3-positions of glycerides, and therefore are suitable for producing the triglyceride of the present invention. Is.

The transesterification reaction product obtained by the above-mentioned various transesterification processes can be used as the triglyceride used in the present invention depending on the kind of raw material selected, but it can be obtained by the above-mentioned chemical synthesis method. As in the case of the esterification product, if necessary, alkali deoxidation treatment, adsorption / fractionation treatment, solvent fractionation treatment or solvent-free fractionation (wintering) treatment, etc. are appropriately combined and subjected to the transesterification reaction product, The triglyceride used in the present invention can be obtained by removing impurities or fractionating or concentrating the glyceride component. The triglyceride is preferably deodorized.

The triglyceride according to the present invention can also be obtained by a method of extracting oils and fats from natural products. That is, among those described as the raw materials for transesterification (component a-1), whales, harbor seals,
harp seal etc.), body tissues of marine mammals such as fur seals, milk secreted from the animals, cells of microalgae such as chlorella, spirulina, and donariella, or cultured cells thereof, genus Nannochloropsis, Trustokis Microalgae belonging to the genus Thraustochytrium, the genus Crypthecodinium and the genus Isochrysis, for example Nannochloropsisoculata
), Trustchytrium aureum (Thraustochytr
ium aureum), Cryptecodynium cornie (Cry
pthecodinium cohnii), Isocrisis galvana (Isoch
rysis galbana) etc. or these cultured cells are used as raw materials. It should be noted that when the origin is a microorganism, the triglyceride obtained therefrom may be any one as long as it satisfies the glyceride structure of the present invention.

These are pressed or n-hexane,
Chloroform, benzene, diethyl ether, an organic solvent such as methanol is extracted or fractionated to obtain an oil, which is subjected to degumming, alkaline deoxidation, decolorization, deodorization, and other free fatty acid, phospholipid, The glyceride fraction can be obtained by removing impurities such as glycolipids, unsaponifiable substances, coloring substances and odorous components. This glyceride fraction can be used as the triglyceride used in the present invention, but the glyceride fraction is further fractionated by solventless low temperature fractionation, solvent fractionation or a silica gel column etc. to bind to the 2-position of triglyceride. It is also possible to produce triglycerides with even fewer long-chain polyunsaturated fatty acid residues.

The triglyceride of the present invention produced by the chemical synthesis method, the transesterification method, the extraction method from a natural product or the like as described above has an n-3 long-chain polyunsaturation as its constituent fatty acid. Less than 40 mol% of the total amount of fatty acids form an ester bond at the 2-position of the triglyceride, but more preferably less than 20 mol%. If it is 40 mol% or more, the desired effect of the present invention becomes small. The triglyceride of the present invention can be used as it is as fats and oils,
It can also be used as a fat or oil by mixing with a common edible fat or oil, for example, the animal or vegetable fat or oil as described as the component b-1. At this time, the content of the triglyceride of the present invention is preferably 5 to 100% by weight of the total fat and oil, and further 10 to
100% by weight is even more preferred. Most preferably 20
１００100% by weight. If it is less than 5% by weight, the desired effect of the present invention is small.

The oil and fat according to the present invention can be ingested in the form of capsules such as soft capsules and microcapsules together with ordinary edible animal and vegetable oils and fats, vitamin E, β-carotene, etc. As a raw material, it can be used as a raw material for various processed foods and as a material for cooking. Alternatively, the oil and fat according to the present invention can be expected to be used for the prevention and treatment of hyperlipidemia.

[0025]

【Example】

Example 1 1 kg of triolein and fish oil (manufactured by Tama Biochemical Co., Ltd., trade name: EPA-18) were hydrolyzed mixed fatty acids at low temperature fractionated fish oil hydrolyzed fatty acid concentrate (C _{20: 5} : 3 in total fatty acids).
7.4 mol%, C22 _{: 5} : 5.4 mol%, C22 _{: 6} : 25.
2 mol%. 72. as an n-3 long-chain polyunsaturated fatty acid.
5 mol%. Add 0.01 wt% BHT. ) With a molar ratio of 1: 5 to adjust the water content to 0.2% by weight, and then use lipozyme IM20 (trade name; lipase derived from Mucor miehei, manufactured by Novo Nordisk). Packed glass column (10cmφ × 60c
m) was allowed to undergo a selective transesterification reaction at 40 ° C.

After removing free fatty acids from the transesterification reaction product by steam distillation and washing with water, a stainless steel column filled with silica gel (Wako Pharmaceutical Co., Ltd., Wakogel C100) infiltrated with n-hexane. The resulting mixture was eluted with n-hexane to remove the diglyceride to obtain 720 g of the triglyceride of the present invention. Total fatty acid composition constituting the triglyceride, 1st and 3rd of glyceride
The fatty acid composition of each of the 2nd and 3rd positions was determined by GLC analysis.
Table 1 shows the results. C that constitutes this triglyceride
90 mol% of _{20: 5} . 95 mol% or more of C _{22: 6} and 93.5 mol% of the total amount of the n-3 long-chain polyunsaturated fatty acids were distributed in the 1st and 3rd positions of the triglyceride. That is, 6.5 mol% of the total amount of the n-3 long-chain polyunsaturated fatty acid was distributed at the 2-position of this triglyceride. This triglyceride was used as a test oil for the following animal experiments.

0.1% by weight of sodium methoxide was added to a part of the present triglyceride, and a random transesterification reaction was carried out at 100 ° C. under reduced pressure, followed by filtration using Celite to obtain a random transesterification product of the present triglyceride. Got The total fatty acid composition, the respective fatty acid compositions at the 1st, 3rd and 2nd positions were determined in the same manner as described above (see Table 1). At the 2-position of this triglyceride, 50.6 mol% of the total amount of the n-3 long-chain polyunsaturated fatty acid was distributed. This random transesterification product was used as a control oil for animal experiments.

[0028]

[Table 1] * Total carbon number: Displayed as the number of double bonds. (N-3) represents an n-3 fatty acid.

Seven 4-week-old male SD rats were used as one test group, and a breeding experiment was carried out using a feed containing 5% by weight of each of the test oil and the control oil (see Table 2). During this period, the feed was prepared and fed daily in order to prevent oxidative deterioration of the feed components. After being allowed to freely ingest water and each of the above-mentioned feeds and bred for 3 weeks, neutral lipids in blood and liver of each test group rat,
The total cholesterol and phospholipid contents were measured.
Table 3 shows the results. In addition, feed intake,
No significant difference was observed in weight gain or liver weight. From this experimental result, the triglyceride according to the present invention (test oil) has no side effect on rats, and in the group to which the test oil was added, blood triglyceride (neutral lipid) and total cholesterol values and liver triglyceride value were It was revealed that the reduction was remarkable.

[0030]

[Table 2] * 1 Nippon Claire Co., Ltd., AIN-93G-MX * 2 Nippon Claire Co., Ltd., AIN-93-VX * 3 t-Butylhydroquinone

[0031]

[Table 3] * There is a significant difference with a risk rate of 5% or less with respect to the value of the control oil added section.

Example 2 A test fat (fat containing a triglyceride of the present invention) and a control fat were prepared as follows. That is, the test fat is h
The arp seal (seal) oil and fat was cooled with a dry ice / acetone refrigerant at -80 ° C for 1 hour, and the precipitated crystal part was filtered off with a filter paper to prepare. The control fats and oils are two types of fish oils with different fatty acid composition (mixed oil of cod liver oil and miscellaneous fish oil, tuna orbital oil), cooled with dry ice / acetone refrigerant in the same manner, and the concentrated fractions are blended to obtain the total fatty acid composition. It is almost similar to that of the test fat. Table 4 shows these fatty acid compositions.

[0033]

[Table 4] *: Same as the notes in Table 1.

Seven 4-week-old SD male rats were used as one test group, and oils and fats containing 20% by weight of each of the above-mentioned test oils and control oils and fats (test oils and control oils and fats 20 parts by weight, palm oil 50 parts by weight, A mixed fat containing 5 parts by weight of oleic safflower oil and 25 parts by weight of high linoleum safflower oil (see Table 5 for the composition of fatty acids), each containing 10% by weight (the composition of the feed is 5% by weight of fat and 10% by weight). , The same as in Example 1 except that 41.7% by weight of corn starch was changed to 36.7% by weight). During this time,
Feed was prepared and fed daily to prevent oxidative degradation of feed ingredients. After free feeding of water and each of the above-mentioned feeds and feeding for 3 weeks, the content of neutral lipid, total cholesterol and phospholipid in blood and liver of each test group rat was measured (see Table 6). No significant difference was observed in feed intake, body weight gain and liver weight in each test group.
From the results of this experiment, it was confirmed that the fats and oils according to the present invention had no adverse effects on rats and effectively reduced the levels of triglyceride (neutral lipid) and total cholesterol in blood and liver.

[0035]

[Table 5] *: Same as the notes in Table 1. (n-6) represents an n-6 fatty acid. * *: Ratio of each fatty acid among saturated fatty acids, monounsaturated fatty acids, n-6 fatty acids and n-3 fatty acids.

[0036]

[Table 6] *: Same as the notes in Table 3.

Example 3 The same experiment as in Example 2 was carried out in the same manner as in Example 2 except that the fats and oils used in Example 2 and the control fats and oils with different mixing ratios were added to the feed. That is, seven 4-week-old SD male rats were used as one test group, and the fats and oils containing 10 wt% of the test fat and oil or the control fat and oil described in Example 2 (10 parts by weight of the test fat and oil and the control oil and fat, 50 parts by weight of palm oil) were used. , A mixed oil of 10 parts by weight of high oleic safflower oil and 30 parts by weight of high linoleum safflower oil. See Table 7 for the fatty acid composition. The same as the above). During this time, the feed was prepared daily to prevent oxidative degradation of the feed components. After free feeding of water and each of the above-mentioned feeds and feeding for 3 weeks, the contents of neutral lipid, total cholesterol and phospholipid in blood and liver of each test group rat were measured (see Table 8). No significant difference was observed in feed intake, body weight gain and liver weight in each test group.
From the results of this experiment and the results of Example 2, the fats and oils according to the present invention have no adverse effect on rats, including the fats and oils containing a small amount of test fats and oils compared to the control fats and oils,
It was found to have an effect of significantly reducing triglyceride (neutral lipid) levels in blood and liver.

[0038]

[Table 7] * And **: Same as the notes in Table 5.

[0039]

[Table 8] *: Same as the notes in Table 3.

Example 4 Microalga Crypthecod
inium cohnii, ATCC 30336) was planted in 30 liters of the medium shown in Table 9, aerobically cultivated at 30 ° C. for 100 hours in a jar fermenter, and cultured alga cells were collected by centrifugation and frozen. Dried (yield 6
25 g). Chloroform: methanol =
Hyscotron (trade name) in a mixed solvent of 1: 1 (weight ratio).
Cell disruption and extraction by Nissin Medical Science Instruments)
520 g of oil was obtained. Silica gel dispersed in n-hexane (manufactured by Wako Pure Chemical Industries, Ltd., trade name: Wakogel C1)
00) packed in a stainless steel column and eluted with diethyl ether: n-hexane = 10: 90 (volume ratio) to give the triglyceride 25 of the present invention.
0 g was obtained. The fatty acid composition of the present triglyceride (this was used as a test fat) was determined in the same manner as in Example 1 (Table 1).
0).

[0041]

[Table 9] (PH: 6.8) * 1: Vitamin mix aqueous solution (unit: weight in 1 liter of the aqueous solution) Biotin: 0.003 g Thiamine: 1.000 g * 2: Metal mix aqueous solution (unit: in 1 liter of the aqueous solution) Weight) Na ₂ EDTA: 1.00 g FeCl ₃ .6H ₂ O: 0.05 g H ₃ BO ₃ : 1.00 g MnCl ₂ .4H ₂ O: 0.15 g ZnCl ₂ : 0.01 g CoCl ₂ · 6H ₂ O: 0.005g

[0042]

[Table 10] *: Same as the notes in Table 1.

The thus-obtained microalgae-derived triglyceride (test oil and fat) and the control oil and fat described in Example 2 at 10% by weight (10 parts by weight of test oil or control oil and fat, 50 parts by weight of palm oil, Hiole) 10 parts by weight of Ick safflower oil and 3 of high linoleum safflower oil
0 parts by weight of mixed fats and oils. See Table 11 for fatty acid composition)
A feed containing 0% by weight (the feed composition is the same as in Example 3 except for fat content) was prepared, and the same breeding test as in Example 3 was performed. Table 12 shows the analysis results of the blood and liver lipid contents of each test group rat. In addition, feed intake,
No significant difference was observed in weight gain or liver weight. From the results of this experiment and the results of Example 2, the fats and oils according to the present invention had no adverse effects on rats, and the fats and oils containing a small amount of test fats and oils in comparison with the control fats and oils were examined in blood and liver. It was found to have the effect of significantly reducing the levels of triglyceride (neutral lipid) and total cholesterol in the medium.

[0044]

[Table 11] * And **: Same as the notes in Table 5.

[0045]

[Table 12] *: Same as Table 3.

[0046]

INDUSTRIAL APPLICABILITY According to the present invention, there is no side effect on animals, and the effect of reducing blood lipid concentration is large as compared with the conventional n-3 long-chain polyunsaturated fatty acid source, Ingesting a smaller amount than conventional sources of n-3 long-chain polyunsaturated fatty acids such as fish oil reduces triglyceride levels and / or cholesterol levels in blood and liver and facilitates blood lipid improvement. It is possible to provide oils and fats with action.

[Procedure amendment]

[Submission date] August 12, 1996

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] 0004

[Correction method] Change

[Correction contents]

On the other hand, eicosapentaenoic acid (all cis-
5,8,11,14,17-eicosapentaenoic acid, hereinafter abbreviated as EPA. The numbers after C _{20: 5} and C represent the total number of carbons: the number of double bonds, and the same shall apply hereinafter. ) And docosahexaenoic acid (all
cis-4,7,10,13,16,19-docosahexaenoic acid, hereinafter abbreviated as DHA. It has been clarified by animal experiments and clinical experiments that n-3 long-chain polyunsaturated fatty acids such as C _{22: 6} ) and food materials containing them have an action of reducing serum triglyceride level and cholesterol level. (For example, J. Dyerberg et al., Prog. Lipid Res., Volume 21, Vol.
255-269, 1982 ). The mechanism of action for reducing the triglyceride level in serum is that the intake of fats and oils containing n-3 long-chain polyunsaturated fatty acids suppresses the triglyceride synthesizing ability in the liver, resulting in the release of triglyceride into the blood. It is presumed that this is due to the suppression of the following (Kenji Hara, Fats and Oils, Vol. 46, No. 4, page 90, 1993)
Year). It is also presumed that the decrease in serum cholesterol level is due to the fact that the n-3 long-chain polyunsaturated fatty acid suppresses the cholesterol synthesizing ability in the liver. (Ch
oi, YS, et al., Lipids, 24, 45, 1989.
Year).

[Procedure amendment 2]

[Document name to be amended] Statement

[Correction target item name] 0016

[Correction method] Change

[Correction contents]

The raw materials for the transesterification are vegetable oils such as linseed oil, sesame oil, perilla oil, sardine oil, cod liver oil, fish oil such as herring oil, squid oil, tuna orbital oil, whales, seals, etc. as component a-1. Compressed or extracted oil obtained from marine mammals such as fur seals, milk fat of the animals, chlorella, spirulina, donaliella , etc., and also genus Nannochloropsis (eg Nannochloropsis oculata , UTEX
LB 2164 etc. ), Trust chytrium (eg Thrausto
chytrium aureum , ATCC 28211, 34304 etc. ), Crypthecodinium cohnii , AT
CC 30021, 30334, 30336, 50052, etc. ), genus Isochrysis (for example, Isochrysis galbana , CCAP927 / 1, UTE
X LB 987, etc. ), oils and fats extracted from microalgae belonging to the genus Mortierella (M.is)
abellina, IFO 6336, 6739, 7873, 7884, ATCC 4
4853) or the like, or a triglyceride of an n-3 long-chain polyunsaturated fatty acid or a mixed fatty acid with the above-mentioned various fatty acids (see paragraph No. 0012) containing the same in an arbitrary ratio. ATCC: American Type Cul
tureCollection (USA), CCAP: Culture Collection o
f Algae and Protozoa (UK), UTEX: Culture Collec
tion of Algae at the University of Texas (US
Country), IFO: Osaka Fermentation Research Institute. Ingredient a-2
As the above, various fatty acids or derivatives thereof described in paragraph 0012 can be used.

[Procedure 3]

[Document name to be amended] Statement

[Name of item to be corrected] 0029

[Correction method] Change

[Correction contents]

Seven 4-week-old male SD rats were used as one test group, and a breeding experiment was conducted using a test oil and a control oil , each containing 5% by weight of each feed (see Table 2). During this time,
Feed was prepared and fed daily to prevent oxidative degradation of feed ingredients. After allowing water and each of the above-mentioned feeds to be freely fed for 3 weeks, the content of neutral lipid, total cholesterol and phospholipid in blood and liver of each test group rat was measured. Table 3 shows the results. No significant difference was observed in feed intake, body weight gain and liver weight in each test plot. From this experimental result, the triglyceride according to the present invention (test oil) has no side effect on rats, and in the group to which the test oil was added, blood triglyceride (neutral lipid) and total cholesterol values and liver triglyceride value were It was revealed that the reduction was remarkable.

[Procedure amendment 4]

[Document name to be amended] Statement

[Correction target item name] 0034

[Correction method] Change

[Correction contents]

Seven 4-week-old SD male rats were used as one test group, and the above-mentioned test fats and control fats and oils containing 20% by weight each (20 parts by weight of test fats or control fats and 50 parts by weight of palm oil) were used. , A mixed fat of 5 parts by weight of high oleic safflower oil and 25 parts by weight of high linoleum safflower oil. See Table 5 for the fatty acid composition. % And the same as in Example 1 except that 41.7% by weight of corn starch was 36.7% by weight).
During this period, the feed was prepared and fed daily in order to prevent oxidative deterioration of the feed components. Free intake of water and each of the above feeds 3
After breeding for a week, the content of neutral lipid, total cholesterol and phospholipid in blood and liver of each test group rat was measured (see Table 6). No significant difference was observed in feed intake, body weight gain and liver weight in each test group. From the results of this experiment, it was confirmed that the fats and oils according to the present invention had no adverse effects on rats and effectively reduced the levels of triglyceride (neutral lipid) and total cholesterol in blood and liver.

[Procedure Amendment 5]

[Document name to be amended] Statement

[Correction target item name] 0043

[Correction method] Change

[Correction contents]

The triglyceride derived from microalgae (test fat and oil) thus obtained and the control fat and oil described in Example 2 were used.
Used, the mixed oil and fat. The fatty acid composition of each 10 wt% including fat (test oils or control oil 10 parts by weight, palm oil 50 parts by weight, high-oleic safflower oil 10 parts by weight of high-linoleic safflower oil 30 parts by weight Table (See 11)
A feed containing 10% by weight of each of the above was prepared (the feed composition is the same as in Example 3 except for fat content), and the same breeding test as in Example 3 was conducted. Table 12 shows the analysis results of the blood and liver lipid contents of each test group rat. No significant difference was observed in feed intake, body weight gain and liver weight in each test plot. From the results of this experiment and the results of Example 2, the fats and oils according to the present invention had no adverse effects on rats, and the fats and oils containing a small amount of test fats and oils in comparison with the control fats and oils were examined in blood and liver. It was found to have the effect of significantly reducing the levels of triglyceride (neutral lipid) and total cholesterol in the medium.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Office reference number FI Technical display location A61K 31/23 ADN A61K 35/12 35/12 35/80 Z 35/80 A23D 9/00 516

Claims (8)

[Claims] 1. An n-3 long-chain polyunsaturated fatty acid is contained as a constituent fatty acid of the glyceride, and less than 40 mol% of the total amount of the n-3 long-chain polyunsaturated fatty acid is bonded to the 2-position of the glyceride. An oil and fat having a function of reducing blood lipid concentration, which is characterized by comprising triglyceride. 2. An oil and fat having the action of reducing the blood lipid concentration, which comprises the triglyceride according to claim 1 in an amount of 5% by weight or more. 3. The n-3 long-chain polyunsaturated fatty acid is one or more selected from the group consisting of α-linolenic acid, arachidonic acid, eicosapentaenoic acid, docosapentaenoic acid and docosahexaenoic acid. The oil and fat according to claim 1 or 2. 4. The oil or fat according to claim 1, wherein the n-3 long-chain polyunsaturated fatty acid is eicosapentaenoic acid and / or docosahexaenoic acid. 5. The oil or fat according to any one of claims 1 to 4, wherein the triglyceride is obtained from a marine mammal or a microalgae, is concentrated, or is transesterified. 6. The oil or fat according to claim 5, wherein the marine mammal is a whale or a seal. 7. The oil or fat according to claim 5, wherein the microalgae belongs to the genus Nannochloropsis, the genus Trustchitrium, the genus Isochrysis or the genus Crypthecodinium. 8. The oil or fat according to claim 1, wherein the triglyceride is produced by a transesterification reaction using a lipase having specificity at the 1- and 3-positions of the glyceride.