JPS62221629A - Production of phospholipid emulsion - Google Patents

Abstract

PURPOSE:To obtain a stable phospholipid emulsion in a high concentration, having a low viscosity, good fluidity and useful for fluid therapy, by removing a neutral lipid of natural lecithin to give an aqueous dispersion of purified phospholipid, adding a fat or oil and protein to the dispersion and homogenizing the resultant blend. CONSTITUTION:A neutral lipid is removed from natural lecithin by treatment with cold acetone, etc., to give purified lecithin containing >=20wt% phosphatidyl choline (hereinafter abbreviated to PC). The resultant purified phospholipid is preferably dispersed in hot water at >=80 deg.C to afford a 10-50wt% aqueous dispersion. 20-100wt% fat or oil, e.g. soybean oil, rapeseed oil, etc., and 0.1-40wt% protein, e.g. sodium caseinate, albumin, etc., heated at >=80 deg.C are added and blended with the above-mentioned dispersion and homogenized under a high pressure to provide the aimed emulsion. The use of the fat or oil provides a dispersion of low viscosity and addition of the protein affords the stable emulsion.

Description

[Detailed Description of the Invention] [Field of Industrial Application] The present invention is directed to a new phospholipid homogenized with good fluidity and low viscosity in order to make it easier for living organisms to ingest phospholipids containing phosphatidylcholine (hereinafter referred to as PC). The present invention relates to a method for producing a lipid emulsion.

[Conventional technology]

It has long been known that natural lecithins, such as soybean lecithin and egg yolk lecithin, used in foods and medicines have surface activity due to the coexistence of hydrophobic groups and parent wood groups in the structure of the various phospholipids they contain. Utilizing this property, it is used as an emulsifier in various industrial fields. Natural lecithin, which is used as an industrial emulsifier, is usually used as a mixture of phospholipid compositions and fats and oils, and has weak W/○ type and ○/W type emulsifying properties.Using this emulsifying property, margarine, It is widely used in food processing such as instant cocoa drinks, pharmaceutical processing such as mosquito repellents, and leather processing.

On the other hand, focusing on the pharmacological effects and value of PC as a food, there is a method of ingesting phospholipids containing PC by eating capsules of phospholipids dissolved in fats and oils. In this method, in order to ingest log PC, 1000m
Even if it is filled into capsules, it is not practical because you will need to take 40 to 50 pieces of egg yolk oil and about 100 pieces of soybean lecithin.Also, egg yolk phospholipids, which are obtained by removing neutral lipids from egg yolk oil, contain highly unsaturated fatty acids. Soybean phospholipids, which are obtained by removing neutral lipids from soybean lecithin, have good storage stability and ease of processing into dosage form, but they contain PC. The amount is 22-24%
To consume 10g of PC, it is necessary to eat about 50g of soybean phospholipid granules and tablets, which is extremely large and difficult to carry.

[Problem that the invention seeks to solve]

Focusing on the pharmacological effects of PC and its value as a food, P.
In order to ingest C as a medicine or food, it is difficult to take it in the above-mentioned phospholipid dosage form, and it is desirable to change the dosage form. Phospholipid emulsions are preferred as such new dosage forms, but natural lecithin has no dispersibility in water and is difficult to form into emulsions. As mentioned above, natural lecithin is used as an emulsifier, but this is limited to emulsifying a large amount of oil, which is the main ingredient, with a small amount of lecithin. It is difficult to form a stable emulsion.

The present invention is intended to solve the above problems, and aims to provide a method for producing a high-concentration phospholipid emulsion that has low viscosity, good fluidity, is stably homogenized, and is easy to ingest.

[Means for solving problems]

The present invention removes neutral lipids from natural lecithin to obtain purified phospholipids containing 20 weight percent or more of phosphatidylcholine, and adds 10 to 50 weight percent aqueous dispersion of this purified phospholipid to
This is a method for producing a phospholipid emulsion, which comprises adding and mixing 20 to 100% by weight of oil and fat and 0.1 to 40% by weight of protein to the purified phospholipid and homogenizing the mixture.

When we investigated the cause of the poor dispersibility of natural phospholipids in water, we found that in natural phospholipids, PC is dissolved in neutral lipids, and PC is coated with neutral lipids.
It was presumed that the water repellency of the neutral lipid deteriorated its dispersibility in water.

Therefore, in the present invention, neutral lipids in natural lecithin that inhibit dispersibility are removed to produce purified phospholipids with high PC concentration,
This is dispersed in a large amount of water to form an aqueous dispersion. This aqueous dispersion has a low viscosity when the phospholipid content is low;
When it is high, the viscosity becomes high, making it unsuitable as an emulsion. However, by adding oil or fat to this aqueous dispersion, a low-viscosity dispersion can be obtained. However, this dispersion has poor emulsion stability and cannot produce a stable emulsion; however, by adding a small amount of protein to it and emulsifying it, a stable emulsion can be obtained.
The viscosity in the high concentration range further decreases.

Natural lecithins used in the present invention include soybean lecithin and egg yolk lecithin. These natural lecithins have a PC content of 15-16% by weight and are mostly neutral lipids. In the present invention, neutral lipids in such natural phospholipids are removed and P
The purified phospholipid has a C content of 20% by weight or more, preferably 50% by weight or more, and more preferably 70% by weight or more.

PC content is measured by the Iastroscan method.

Known methods such as cold acetone treatment and aqueous ethanol treatment can be used to remove neutral lipids from natural lecithin, and if the target PC content is not reached in one treatment, the same operation can be repeated. or in combination with other operations.

The purified phospholipids thus obtained are dispersed in water using an emulsifying device, stirred until the solid content is completely eliminated, and pre-emulsified.
An aqueous dispersion of At this time, it is preferable to use hot water of 80° C. or higher as a dispersion medium, thereby making it possible to solubilize the fats and oils and proteins added later. If the purified phospholipid concentration in the aqueous dispersion is less than 10% by weight, the final emulsion will have a low active ingredient content, and if it exceeds 50% by weight, the viscosity of the emulsion will increase, resulting in poor emulsification, which is not preferred.

Next, at a temperature of 80° C. or higher, fats and oils heated to the same <80° C. or higher are added little by little to the pre-emulsified aqueous dispersion, dispersed, and refined to form an emulsion. By performing this treatment, the viscosity can be lowered compared to a pre-emulsified liquid having the same purified phospholipid concentration, and the fluidity can be improved.

The fats and oils to be added are preferably soybean oil, rapeseed oil, cottonseed oil, chili pepper oil, safflower oil, etc., which are liquid at room temperature, and the amount added is 20 to 100% by weight based on the purified phospholipid, preferably 5% by weight.
It is 0 to 100% by weight. If the amount of oil 1 is less than 20% by weight, the viscosity lowering effect will be small, and if it exceeds 100% by weight, the PC amount will be relatively reduced, which is not preferable.

Next, the protein is added little by little to this emulsion while stirring while heating to 80° C. or higher, and the protein is dispersed until it is completely dissolved.The protein is further emulsified using a high-pressure dispersion machine to form an emulsion. This treatment stabilizes the emulsion.

It is possible to prevent separation of the oil layer and further reduce the viscosity in the high concentration range.

The proteins to be added include casein, casein sodium salt,
Albumin, egg white, skim milk powder, etc. are added in an amount of 0.1 to 40% by weight, preferably 0.5 to IO% by weight, based on the purified phospholipid. If the amount added is less than 0.1% by weight, there is no emulsion stabilizing effect, and if it exceeds 40% by weight,
The PC content of the emulsion decreases and the viscosity increases.

Homogenization using a high-pressure dispersion machine is carried out under high pressure of 20 to 200 kg/cd using a dispersion machine such as a homogenizer, with the first stage being 50 kg/cd and the second stage being 150 kg/a+.
It is preferable to carry out the dispersion under a high pressure of 100 degrees.

The manufacturing method of the present invention will be explained in more detail below based on experimental results.

Natural lecithins such as soybean lecithin and egg yolk lecithin, which are used as emulsifiers for fat emulsions and contain neutral lipids, can cause phospholipids to aggregate in the upper part of the water layer even after mechanical stirring for preliminary emulsification. There is. This condition is very similar to the feathering phenomenon caused by whitener added to coffee; a completely uniform suspension is not obtained, and oil layer separation is clear. This phenomenon is presumed to have occurred because the phospholipids cannot come into contact with water due to the water repellency of the neutral lipid group such as oil, which is the solvent that solubilizes the phospholipids.

When such natural phospholipids are subjected to cold acetone treatment, hydrous ethanol treatment, etc. to remove neutral lipids, purified phospholipids with a high PC content can be obtained. For example, purified phospholipid (hereinafter referred to as 5-PC-70) with a PC content of 70% (weight% or less) obtained by removing neutral lipids from soybean lecithin is a transparent yellow solid with extremely high plasticity. Its plasticity does not change significantly with cooling or heating, and its taste is mild, but its plasticity makes it difficult to chew with the teeth, and it sticks to the teeth and upper jaw, making it feel like eating clay. difficult.

Similarly, purified phospholipid with a PC content of 70% (hereinafter referred to as E-PC-70), which is obtained by removing neutral lipids from egg yolk lecithin, is brown and translucent, has an oily texture, is soft, and is highly plastic. It is a semi-solid, its plasticity does not change significantly when cooled or heated, it has a very oily taste, the whole mouth remains coated in oil for a long time, and it is difficult to chew, similar to soybean products. It is beautiful to take for a long time because of its dots and stickiness.

According to the results of a preliminary emulsification experiment on the above S-PC-70, E-PC-70, and purified phospholipid with a PC content of 24% (hereinafter referred to as S-PC-24) obtained by removing neutral lipids from soybean lecithin, For all samples, uniform aqueous dispersions up to 10% (vt/wt) were obtained, but aqueous dispersions of 10% or more significantly increased viscosity as shown in Table 1 below. The tendency of viscosity increase is pc
In the case of S-PC-24, which has a low PC content, it shifts to a higher concentration side than 5-PC-70 and E-PC-70, which have a high PC content.

In order to prepare an emulsion containing 10% or more of purified phospholipids with a high PC content and excellent stability and fluidity, it is necessary to add triazyl, which is liquid at room temperature, to a phospholipid In solution that is evenly suspended in water. This is possible by adding glycerol (soybean oil was used in this experiment) and emulsifying it, and as shown in Table 3 below, the viscosity of a 70% PC solution decreases in a 10% solution, and the fluidity decreases. Improved.

Furthermore, in order to reduce the viscosity and improve the stability in the high a degree range, these improvements can be made by adding protein to the above emulsion and coating it with triazylglycerol and phospholipid particles. be. These series of phenomena are caused by the localization of phospholipids around a neutral lipid core, and lipid carriers in the blood that are coated with proteins to maintain the stability of oil droplets in water. It is assumed that the structure is similar to that of lipoproteins.

Triazylglycerol added phospholipid emulsion.

Even if emulsification is performed, oil layer separation is observed in the upper layer of the stored sample after about a week, but it is possible to maintain the stability of the oil droplets by adding protein.

To prepare a protein-containing emulsion, protein is added to a pre-emulsion of phospholipid added with triazylglycerol, sufficiently solubilized by heating and stirring, and then refined and emulsified using a homogenizer. If the emulsion stability of this emulsion is 1, no oil layer separation will be observed even after storage for 1 month.

The viscosity of a typical emulsion in a three-component system of phospholipids, triadylglycerol, and sodium casein salt is shown in Table 1 below, and the viscosity of a refined emulsion with protein addition is shown in Table 2. As is clear from the comparison, PC decreases significantly in solutions with a concentration of 10% or more.

These emulsion systems exhibit characteristic viscosity behavior 810.

That is, as shown in Table 2, a suspension of S-PC-70 alone has a viscosity of 27.380°70 at 80°C when its content increases to 5%, 10%, and 15%.
00cP, but at 15°C, the viscosity increases to 70.2700°40000cP. However, in the emulsion system containing triazylglycerol, as shown in Table 3, 8
The viscosity is 5,17.2500 cP at 0°C, but at 15°C
8, 27.4700 cP, with little tendency to increase viscosity. This is presumed to be because the state of dispersion in the aqueous phase changed from hydration to emulsion.

In addition, in the emulsion system containing sodium caseinate, the first
As shown in the table, the phospholipid content is 5%, 10%,
When the amount increases to 15%, the viscosity of the system at 80°C is
4.38. 80 cPY,,'>ru, intestine ■■■ Garden I-The above is based on emulsification system using other phospholipids, neutral lipids, and proteins. The same is true when albumin, which specifically binds free fatty acids, is supplied from egg white.

The emulsion with a high phospholipid content obtained as described above can be widely applied to infusion therapy. In particular, a phospholipid emulsion with a high PC content is a form suitable for oral ingestion of PC, and is easier to eat and ingest in large quantities than solid PC. Another advantage is that even if you take in 20g of PC per day, you only need to take in about 500cal of energy.

Soybean lecithin and egg yolk lecithin are substances with good biocompatibility and low toxicity, as they are used in fat emulsions, especially indoralipid and microsphere emulsifiers.For example, E-PC-70 and S-PC-70 are Sutra Lochi LD
Go is 12000mg/kg or more, intravenous injection LD!
. If the amount is 2000 mg/kg or more and triazylglycerol is added, PC74 can be administered orally or via peripheral veins for a short period of time after preparation. Emulsions prepared by pre-emulsifying with phospholipids and triazylglycerol and further adding proteins can also be administered orally, enterally or through other tubes. If a non-antigenic substance such as human plasma albumin is used as the protein used in this system, intravenous injection becomes possible.

〔Effect of the invention〕

As described above, according to the present invention, the neutral lipids of natural lecithin are removed to form an aqueous dispersion, and oil and fat and protein are added to this to emulsify it, resulting in low viscosity, good fluidity, and stability. It is possible to easily produce a highly concentrated phospholipid emulsion that is homogenized and safe to ingest.

〔Example〕

Hereinafter, the present invention will be explained in further detail with reference to Examples and Comparative Examples. In each example, 51 parts indicate weight % and parts by weight, respectively. Further, purified phospholipids were produced as follows.

(2) Soybean lecithin from which neutral fats were removed was treated with aqueous ethanol to obtain 5-PC-70 with a PC content of 70%.

■ Repeated cold acetone treatment of soybean lecithin.

S-PC-24 with a PC content of 24% was obtained.

(2) Egg yolk lecithin was repeatedly treated with cold acetone to obtain E-PC-70 with a PC content of 70%.

Example 1 Using 5-PC-70 and IE-PC-70, an autohomo mixer (manufactured by Tokushu Kika Kogyo) was used as the emulsifying device, and the number of revolutions was 1.
Mechanical emulsification was performed for 30 minutes at 1000 Orp and a temperature of 80°C, and the phospholipid content was 5%, 7.5%, 10%, 15%,
20% and 25% aqueous pre-emulsions were prepared, and 100 parts of soybean oil heated to 80°C was added to the aqueous pre-emulsions based on 100 parts of phospholipids in the aqueous pre-emulsions, and autohomogenized. The mixture was emulsified using a mixer to obtain each emulsion. Add 4 parts to this emulsion per 100 parts of phospholipid in the emulsion.
Add 0 parts of casein sodium salt and use a homogenizer (
An emulsion was obtained by emulsifying the emulsion using a MM type manufactured by Sansho Tokushu Kikai Co., Ltd.).

Table 1 shows the viscosity of the emulsion obtained.

Table 1 Viscosity (cP)
From the results in Table 1, it can be seen that the emulsion with a phospholipid content of 25% has a component concentration of 60%.

The viscosity at 80°C was low and the stability was excellent, and no oil layer separation was observed even after being left at 37°C for one month.

Example 2 Hot water of 80℃ or higher 950℃ in a 20Q stainless steel beaker
Weigh 0 parts and rotate at 240 rpm with a three-one motor.
While stirring with II, 5000 parts of finely sheared 5-PC-70 (described above) was added little by little until the entire mixture became a uniform paste and no solid content was present.

Refined soybean oil heated to 80℃ or higher in a paste-like preliminary emulsion 5
000 parts were added little by little, stirring was continued at the highest stirring speed until the solution changed from a translucent solution to a milky white solution. 50 parts of caseinate sodium salt was added little by little to this emulsion and stirred until completely dissolved. This emulsion was emulsified using a two-stage homogenizer (three-phase special machine IIMM type) at a rate of 50 kg/cd in the first stage and 150 kg/cd in the second stage, and then placed in an autoclave at a temperature of 110°C and a pressure of 2 kg/cd.
It was sterilized for 30 minutes with aJ to obtain an emulsion.

The sterilized emulsion was rapidly cooled to around 15°C, stored in a refrigerator day and night, and then placed in a sterile nitrogen-substituted stoppered flask and stored in a constant temperature bath at 37°C. This emulsion was completely dispersed in water, and the particle size of the emulsion was 3 to 10 μm. The viscosity measured with a B-type rotational viscometer (Tokyo Keiki Seisakusho! I) was 220c at 80°C.
P, 380 cP at 37°C and 730 cP at 15°C. After 1 month in the constant temperature bath, no putrefaction or oil layer separation was observed in the emulsion, but the viscosity at 37°C increased to 640 cP. When the emulsion was orally ingested after one month, it felt somewhat viscous and felt like heavy cream, and it was very easy to swallow 100 ml.

Example 3 Hot water 804 at 80°C or higher in a 20Q stainless steel beaker
Weigh 0 parts and rotate at 240 rpm with a three-one motor.
Add 4,000 parts of E-PC-70 (described above), which is a dry clay-like material that crumbles easily, little by little while stirring at +m, and continue stirring until the entire solution becomes a weak, homogeneous gel-like solution and no yellow solid content is present. continued. 4,000 parts of refined soybean oil heated to 80° C. or higher was added little by little to the gel-like preliminary emulsion, and stirring was continued at the highest stirring speed until the solution changed from a translucent solution to a milky white solution. 40 parts of caseinate sodium salt was added little by little to this emulsion and stirred until completely dissolved.

This emulsion was emulsified using a two-stage homogenizer (described above) at a rate of 50 kg/a# in the first stage and 150 kg/d in the second stage, and then placed in an autoclave at a temperature of 110°C and a pressure of 2 kg/d.
The emulsion was sterilized for 30 minutes at J. The sterilized emulsion was rapidly cooled to around 15°C, stored in a refrigerator day and night, and then placed in a sterile nitrogen-substituted stoppered flask and stored in a constant temperature bath at 37°C. This emulsion was completely dispersed in water, the particle size of the emulsion was 2 to 8 μ-, and the viscosity was 360 cP at 80°C and 520 cP at 37°C.
P, 860 cP at 15°C. No rotting or oil layer separation was observed in the emulsion after one month in the constant temperature bath, but at 37°C.
The viscosity at was increased to 900 cP. After one month, the emulsion was taken orally and only felt a little viscous.

It had a boiled egg-like odor and a coffee cream-like feel, and it was very easy to pour 100 mQ of hot water into a 20 Q stainless steel beaker at 950 °C or higher.
Weigh 0 parts and rotate at 24 Orp using a three-one motor.
4000 parts of finely sheared 5-PC-70 (mentioned above) was added little by little while stirring at m, and the process was continued until the whole became a uniform mesh and no solid content was left. 4,000 parts of purified safflower oil heated to 80° C. or higher was added little by little to the paste-like preliminary emulsion, and stirring was continued at the highest stirring speed until the solution changed from a translucent solution to a milky white solution. 20 parts of albumin was added little by little to this emulsion and stirred until completely dissolved. This emulsion was emulsified using a two-stage homogenizer (described above) at a rate of 50 kg/d in the first stage and 150 kg/d in the second stage, and then placed in an autoclave and sterilized at a temperature of 110°C and a pressure of 2 kg/aJ for 30 minutes to form an emulsion. I got it. The sterilized emulsion was rapidly cooled to around 15°C and stored in a refrigerator overnight, then placed in a sterile nitrogen-substituted stoppered flask and stored in a constant temperature bath at 37°C. This emulsion was completely dispersed in water, and the particle size of the emulsion was 2 to 10 μm. Viscosity is 200cP at 80℃,
It was 320 cP at 37°C and 600 cP at 15°C. After 1 month in the constant temperature bath, no putrefaction or oil layer separation was observed in the emulsion, but the viscosity at 37°C increased to 700 cP. Oral ingestion of the emulsion after 1 month feels slightly viscous.

It had a heavy cream-like feel and was very easy to cover with 100mff1.

Example 5 Hot water 804 at 80°C or higher in a 20Q stainless steel beaker
Weigh 0 parts and rotate at 240 rpm with a three-one motor.
While stirring in the intestines, 000 parts of E-PC-705, which is a dry clay-like material that crumbles easily, was added little by little, and stirring was continued until the entire solution was a weak, homogeneous gel-like solution and no yellow solid content was present. 5,000 parts of purified safflower oil heated to 80° C. or higher was added little by little to the gel-like preliminary emulsion, and stirring was continued at the highest stirring speed until the solution changed from a translucent solution to a milky white solution. Albumin 25% is added to this emulsion.
portion was added little by little and stirred until completely dissolved. This emulsion was passed through a two-stage homogenizer (described above), and the first stage
After emulsifying at 0 kg/d and 150 kg/aJ in the second stage,
Place in an autoclave at a temperature of 110℃ and a pressure of 2kg/aJ.
The mixture was sterilized for 30 minutes to obtain an emulsion. The sterilized emulsion was rapidly cooled to around 15°C and stored in a refrigerator overnight, then placed in a sterile nitrogen-substituted stoppered flask and stored in a constant temperature bath at 37°C. This emulsion was completely dispersed in water, and the particle size of the emulsion was 3 to 10 microns. Viscosity is 400cP at 80℃ and 550c at 37℃
P, 940 cP at 15°C. No rotting or oil layer separation was observed in the emulsion after one month in the constant temperature bath, but at 37°C.
The viscosity at was increased to 1000 cP. When the emulsion was orally ingested after one month, the emulsion felt slightly viscous and had a coffee cream-like texture, and it was very easy to pour over 100 mjl.

Comparative Example 1 Using 5-PC-24, [-PC-70, and S-PC-70, the autohomogen mixer (described above) was used as an emulsifying device, and the rotation speed was too high.

Mechanical emulsification was performed at rpm and temperature of 80°C for 30 minutes, and the phospholipid content was 5%, 10%, 12%, and 15%, respectively.

A 17% aqueous preemulsion was obtained. The viscosity of the obtained preliminary emulsion is shown in Table 2.

Table 2 Viscosity (c
P) Note - indicates not measurable.

A homogeneous emulsion can be obtained in both cases, but the viscosity increases significantly from 10% or more.
Shifts to a higher concentration side than C-70.

Comparative Example 2 Na2. Add the same amount of soybean oil as the phospholipids heated to 80°C to the pre-emulsified liquid of &3, and emulsify using a homogenizer (described above) to obtain phospholipid contents of 5%, 10%, and 12%.
%, 15%, and 17% emulsions were obtained. The viscosity of the obtained emulsion is shown in Table 3.

Table 3 Viscosity (cP)
The viscosity was significantly lower than the results of Comparative Example 1, and the viscosity at 15°C was also low, but separation of the oil layer was observed in one week.

Claims (5)

[Claims] (1) Neutral lipids are removed from natural lecithin to obtain a purified phospholipid containing 20% by weight or more of phosphatidylcholine, and a 10 to 50% by weight aqueous dispersion of this purified phospholipid is added to A method for producing a phospholipid emulsion, which comprises adding and mixing 100% by weight of oil and fat and 0.1 to 40% by weight of protein and homogenizing the mixture. (2) The method according to claim 1, wherein the aqueous dispersion is a dispersion of purified phospholipids in hot water of 80° C. or higher. (3) The method according to claim 1 or 2, wherein the fat or oil is a liquid oil. (4) The method according to any one of claims 1 to 3, wherein the protein is casein sodium salt or albumin. (5) The method according to any one of claims 1 to 4, wherein the homogenization is performed under high pressure.