Egg yolk phospholipid composition, method for producing same, and fat emulsion and lipolytic preparation using said egg yolk phospholipid composition

Abstract

A yolk phospholipid composition having a lysophosphatidyl ethanolamine content of 1% by mass or less and an amino acid content of 50 mg / 100 g or more and 500 mg / 100 g or less.

Description

[0001] This application is a divisional application of the application filed on August 31, 2016, with application number 201680088824.6 and entitled "Egg Yolk Phospholipid Composition and Method for Manufacturing the Same Thereof, and Fat Emulsion and Lipolysis Preparation Using the Egg Yolk Phospholipid Composition". Technical Field

[0002] This invention relates to egg yolk phospholipid compositions and methods for their manufacture, as well as fat emulsions and lipolysis preparations using egg yolk phospholipid compositions. Background Technology

[0003] Egg yolk phospholipids are widely used as emulsifiers in fields such as cosmetics and pharmaceuticals, including fat emulsions and lipolysis preparations (Patent Document 1). In recent years, there has been a search for emulsifiers using egg yolk phospholipids that are of high purity and exhibit excellent stability.

[0004] Existing technical documents

[0005] Patent documents

[0006] Patent Document 1: International Publication No. WO2009 / 093650 Summary of the Invention

[0007] The problem the invention aims to solve

[0008] This invention provides an egg yolk phospholipid composition for manufacturing high-purity and highly stable emulsions, a method for manufacturing the same, and fat emulsions and lipolysis preparations using the egg yolk phospholipid composition.

[0009] Solution for solving the problem

[0010] The inventors of this application discovered that by adjusting the content of lysophosphatidylethanolamine and amino acids in egg yolk phospholipids to a specified amount, a high-purity emulsion with excellent stability can be produced, thus completing this invention.

[0011] 1. An egg yolk phospholipid composition of the present invention, wherein the content of lysophosphatidylethanolamine is less than 1% by mass and the content of amino acids is more than 50 mg / 100g and less than 500 mg / 100g.

[0012] 2. The egg yolk phospholipid composition according to 1 above, wherein when the total amount of the amino acid is set to 100 parts by mass, the total proportion of serine and threonine in the amino acid can be 10 parts by mass or more.

[0013] 3. The egg yolk phospholipid composition according to 1 or 2 above, wherein when the total amount of the amino acid is set to 100 parts by mass, the total proportion of serine, threonine, lysine and arginine in the amino acid can be more than 10 parts by mass.

[0014] 4. The egg yolk phospholipid composition according to any one of 1 to 3 above, wherein when the total amount of the egg yolk phospholipid is set to 100 parts by mass, the content of the amino acid relative to the content of the egg yolk phospholipid can be more than 0.06 parts by mass and less than 0.6 parts by mass.

[0015] 5. The egg yolk phospholipid composition according to any one of 1 to 4 above, wherein it may contain phosphatidylcholine, and when the content of the phosphatidylcholine is set to 100 parts by weight, the content of the amino acid is 0.1 parts by weight or more and 0.8 parts by weight or less relative to the phosphatidylcholine.

[0016] 6. The egg yolk phospholipid composition according to any one of 1 to 5 above, wherein it may contain phosphatidylethanolamine, and when the content of the phosphatidylethanolamine is set to 100 parts by weight, the content of the above-mentioned amino acid is 0.2 parts by weight or more and 10 parts by weight or less relative to the content of the phosphatidylethanolamine.

[0017] 7. The egg yolk phospholipid composition according to any one of 1 to 6 above, wherein it may contain 65% by mass or more of phosphatidylcholine and 5% by mass or more of phosphatidylethanolamine.

[0018] 8. The egg yolk phospholipid composition according to any one of 1 to 7 above, wherein it may contain phosphatidylethanolamine, and when the content of the phosphatidylethanolamine is set to 100 parts by mass, the content of the lysophosphatidylethanolamine is 3 parts by mass or more and 15 parts by mass or less relative to the content of the phosphatidylethanolamine.

[0019] 9. The egg yolk phospholipid composition according to any one of 1 to 8 above, wherein the cholesterol content may be less than 1.5% by mass.

[0020] 10. The egg yolk phospholipid composition according to any one of 1 to 9 above, wherein the content of free fatty acids may be less than 1.0% by mass.

[0021] 11. A fat emulsion according to one aspect of the present invention, comprising: emulsified particles containing the egg yolk phospholipid composition described in any one of 1 to 10 above, and an aqueous phase in which the emulsified particles are dispersed.

[0022] 12. A lipolysis preparation according to one aspect of the present invention comprises: emulsified particles containing the egg yolk phospholipid composition and the agent described in any one of 1 to 10 above, and an aqueous phase in which the emulsified particles are dispersed.

[0023] 13. A method for manufacturing an egg yolk phospholipid composition according to one aspect of the present invention, comprising the following steps: mixing a first egg yolk phospholipid composition and a second egg yolk phospholipid composition to obtain a final egg yolk phospholipid composition, wherein the first egg yolk phospholipid composition has a lysophosphatidylethanolamine content of 1% by mass or less and an amino acid content of 0 mg / 100g or more and 250 mg / 100g or less, the second egg yolk phospholipid composition has a lysophosphatidylethanolamine content of more than 1% by mass and an amino acid content of 250 mg / 100g or more and 500 mg / 100g or less, and the final egg yolk phospholipid composition has a lysophosphatidylethanolamine content of 1% by mass or less and an amino acid content of 50 mg / 100g or more and 700 mg / 100g or less.

[0024] 14. A method for manufacturing an egg yolk phospholipid composition according to one aspect of the present invention, comprising the following steps: a first resin treatment step, wherein a first raw material egg yolk phospholipid composition having a lysophosphatidylethanolamine content of more than 1% by mass and less than 3% by mass, and an amino acid content of more than 500 mg / 100g is treated with a first ion exchange resin, thereby obtaining a first egg yolk phospholipid composition having a lysophosphatidylethanolamine content of less than 1% by mass and an amino acid content of more than 0 mg / 100g and less than 250 mg / 100g; a second resin treatment step, wherein a first raw material egg yolk phospholipid composition having a lysophosphatidylethanolamine content of more than 1% by mass and less than 3% by mass, and an amino acid content of more than 500 mg / 100g is treated with a second ion exchange resin; The process involves treating a second raw material egg yolk phospholipid composition having a content of 1% to 3% by mass and an amino acid content of more than 500 mg / 100g with resin to obtain a second egg yolk phospholipid composition having a lysophosphatidylethanolamine content of more than 1% by mass and an amino acid content of more than 250 mg / 100g and less than 500 mg / 100g; and mixing the first egg yolk phospholipid composition with the second egg yolk phospholipid composition to obtain a final egg yolk phospholipid composition having a lysophosphatidylethanolamine content of less than 1% by mass and an amino acid content of more than 50 mg / 100g and less than 500 mg / 100g.

[0025] 15. The method for manufacturing the egg yolk phospholipid composition according to 14 above, wherein, in the first resin treatment step, the rate of decrease of the amino acid content in the egg yolk phospholipid composition per minute is 100 mg / 100g or more and 200 mg / 100g or less, and in the second resin treatment step, the rate of decrease of the amino acid content in the egg yolk phospholipid composition per minute is 5 mg / 100g or more and 30 mg / 100g or less.

[0026] 16. The method for manufacturing the egg yolk phospholipid composition according to 14 or 15 above, wherein, in the mixing step, the mass ratio of the first egg yolk phospholipid composition to the second egg yolk phospholipid composition can be 0.2 or more and 5 or less.

[0027] The effects of the invention

[0028] According to any one of the egg yolk phospholipid compositions described in any of the above 1 to 10, by making the content of lysophosphatidylethanolamine 1% or less by mass and the content of amino acids 50 mg / 100g or more and 500 mg / 100g or less, the amount of amino acids in the egg yolk phospholipid is adjusted to the above range, and therefore, using the egg yolk phospholipid composition, a high-purity emulsion with excellent stability can be produced.

[0029] The fat emulsion described in 11 above comprises emulsified particles containing the egg yolk phospholipid composition described in any one of 1 to 10 above, and an aqueous phase in which the emulsified particles are dispersed, thereby achieving high purity and excellent emulsification stability.

[0030] The lipolysis preparation described in 12 above comprises emulsified particles containing the egg yolk phospholipid composition and the agent described in any one of 1 to 10 above, and an aqueous phase in which the emulsified particles are dispersed, thereby achieving high purity and excellent emulsification stability.

[0031] According to the method for manufacturing the egg yolk phospholipid composition described in 13 above, by mixing the first egg yolk phospholipid composition and the second egg yolk phospholipid composition, a final egg yolk phospholipid composition having a lysophosphatidylethanolamine content of 1% by mass or less and an amino acid content of 50 mg / 100g or more and 700 mg / 100g or less can be obtained in a simple and efficient manner.

[0032] According to the method for manufacturing the egg yolk phospholipid composition according to any one of the above 14 to 16, by including the above-described first resin treatment step, the above-described second resin treatment step, and the step of obtaining the above-described final egg yolk phospholipid composition, an egg yolk phospholipid composition having a lysophosphatidylethanolamine content of 1% by mass or less and an amino acid content of 50 mg / 100g or more and 500 mg / 100g or less can be manufactured in a simple and efficient manner. Detailed Implementation

[0033] It should be noted that, unless otherwise stated, in this invention, "parts" means "parts by mass" and "%" means "% by mass".

[0034] The egg yolk phospholipid composition of one embodiment of the present invention has a lysophosphatidylethanolamine (hereinafter also referred to as "LPE") content of 1% by mass or less, and an amino acid content of 50 mg / 100g or more and 500 mg / 100g or less.

[0035] According to the egg yolk phospholipid composition of this embodiment, the LPE content is 1% by mass or less, and the amino acid content is 50 mg / 100g or more and 500 mg / 100g or less, thereby using the egg yolk phospholipid composition can produce emulsions with high purity and excellent stability.

[0036] The inventors have discovered that the pH decrease that occurs during high-temperature sterilization of the emulsion obtained using the egg yolk phospholipid composition of this embodiment is due to the amount of amino acids removed along with LPE during LPE removal. This pH decrease is one of the factors that reduces the stability of the emulsion.

[0037] More specifically, the LPE content of the egg yolk phospholipid composition of this embodiment is suppressed to 1% by mass or less, and the amino acid content is adjusted to 50 mg / 100g or more and 500 mg / 100g or less. Therefore, the emulsion using the egg yolk phospholipid composition of this embodiment has high purity, and even after high-temperature treatment during sterilization, the pH of the emulsion decreases little, and the stability is also excellent.

[0038] (Egg yolk phospholipid composition)

[0039] In this invention, "egg yolk phospholipid composition" refers to a composition containing phospholipids derived from egg yolk.

[0040] (phospholipids)

[0041] Examples of phospholipids include: phosphatidylcholine (hereinafter also referred to as "PC"), phosphatidylethanolamine (hereinafter also referred to as "PE"), phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, phosphatidyl glycol and phosphatidyl polyethylene glycol, phosphatidic acid, lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylglycerol, their hemolysates, and sphingomyelin, which may contain one or more of these. These are all phospholipids derived from egg yolks.

[0042] The egg yolk phospholipid content (total) in the egg yolk phospholipid composition of this embodiment is 65% by mass or more and less than 100% by mass, typically 99.95% by mass or less, and preferably 75% by mass or more from the viewpoint that it can produce an emulsion with better stability; on the other hand, it is 99.5% by mass or less, and more preferably 95% by mass or less.

[0043] In this embodiment, the egg yolk phospholipid composition may contain, for example, 65% by mass or more of PC (preferably 68% by mass or more, more preferably 70% by mass or more; and preferably 99% by mass or less, more preferably 95% by mass or less, and more preferably 90% by mass or less), and 5% by mass or more of PE (preferably 7% by mass or more, more preferably 10% by mass or more; and preferably 20% by mass or less).

[0044] The LPE content in the egg yolk phospholipid composition of this embodiment is 1% by mass or less, and can be 0.8% by mass or less. By keeping the LPE content below the above-mentioned upper limit, a high-purity and highly stable egg yolk phospholipid composition can be obtained. The lower limit of the LPE content is not particularly limited, and a highly stable egg yolk phospholipid composition can be easily obtained by keeping it at 0.01% by mass or more, and further at 0.1% by mass or more.

[0045] Furthermore, from the perspective of superior stability, the egg yolk phospholipid composition of this embodiment contains PE. When the content of PE is set to 100 parts by mass, the content of LPE is preferably 2 parts by mass or more and 15 parts by mass or less relative to the content of PE, more preferably 3 parts by mass or more, and more preferably 10 parts by mass or less.

[0046] (Amino acids)

[0047] In the case where the egg yolk phospholipid composition of this embodiment contains two or more amino acids, the content of the amino acids is the total amount of the two or more amino acids. In the egg yolk phospholipid composition of this embodiment, from the viewpoint of further improving stability, the content (total amount) of amino acids is preferably 50 mg / 100g or more; furthermore, from the viewpoint of further improving the purity of the phospholipid composition, it is more preferably 500 mg / 100g or less; on the other hand, it is preferably 100 mg / 100g or less, and more preferably 400 mg / 100g or less.

[0048] Examples of amino acids include: essential amino acids (tryptophan, lysine, methionine, phenylalanine, threonine, valine, leucine, isoleucine, histidine), non-essential amino acids (arginine, cysteine, tyrosine, glycine, alanine, serine, cysteine, asparagine, glutamine, proline, aspartic acid, glutamic acid), or amino acids other than those listed above. From the viewpoint of producing emulsions with superior stability, it is preferable to contain at least one selected from serine, threonine, lysine, and arginine, and more preferably, serine and / or threonine.

[0049] In the egg yolk phospholipid composition of this embodiment, from the viewpoint of producing an emulsion with superior stability, when the total amount of the above-mentioned amino acids is set to 100 parts by mass, the combined ratio of serine and threonine in these amino acids is more preferably 10 parts by mass or more, and even more preferably 20 parts by mass or more. On the other hand, there is no particular upper limit to the combined ratio of serine and threonine in these amino acids; for example, from the perspective of avoiding the difficulty in obtaining an effect that matches the content, it can be set to 50 parts by mass or less.

[0050] Furthermore, in the egg yolk phospholipid composition of this embodiment, from the viewpoint of producing an emulsion with superior stability, when the total amount of the above-mentioned amino acids is set to 100 parts by mass, the combined proportion of serine, threonine, lysine, and arginine in these amino acids is preferably 10 parts by mass or more, more preferably 15 parts by mass or more, and even more preferably 20 parts by mass or more. On the other hand, there is no particular upper limit to the combined proportion of serine, threonine, lysine, and arginine in these amino acids, but from the perspective of avoiding the difficulty in obtaining an effect that matches the content, it is preferably 70 parts by mass or less.

[0051] From the viewpoint of being able to produce emulsions with better stability, in the egg yolk phospholipid composition of this embodiment, when the content of PC is set to 100 parts by mass, the content of the above-mentioned amino acids is preferably 0.1 parts by mass or more and 0.8 parts by mass or less relative to the content of PC, more preferably 0.2 parts by mass or more; on the other hand, it is more preferably 0.7 parts by mass or less.

[0052] From the perspective of superior stability, the egg yolk phospholipid composition of this embodiment contains PE. When the content of PE is set to 100 parts by mass, the content of the above-mentioned amino acids is preferably 0.2 parts by mass or more and 10 parts by mass or less relative to the content of PE, more preferably 0.5 parts by mass or more, and more preferably 5 parts by mass or less.

[0053] Furthermore, from the viewpoint of being able to produce an emulsion with superior stability, in the egg yolk phospholipid composition of this embodiment, when the content of the above-mentioned egg yolk phospholipid (total) is set to 100 parts by mass, the content of the above-mentioned amino acids relative to the content of the egg yolk phospholipid (total) is preferably 0.06 parts by mass or more and 0.6 parts by mass or less, more preferably 0.1 parts by mass or more, and more preferably 0.5 parts by mass or less.

[0054] (cholesterol)

[0055] In the egg yolk phospholipid composition of this embodiment, from the viewpoint of producing an emulsion with better stability, the content of cholesterol (hereinafter also referred to as "CH") can be 1.5% by mass or less, preferably 1.0% by mass or less, more preferably 0.8% by mass or less; on the other hand, it can be set to 0.01% by mass or more.

[0056] In this invention, the cholesterol content is a value determined using the following method.

[0057] Weigh 0.5 g of sample into a 10 mL stoppered test tube, dissolve in chloroform, and bring the volume to 10 mL. Develop and measure the sample using an IATROSCANTH-10 according to the operating procedure. Since the sensitivity varies depending on the lipid composition, for triglycerides and cholesterol, the area value (AREA) is multiplied by the following coefficient to obtain the corrected area value.

[0058] Triglycerides × 0.9

[0059] Cholesterol × 0.5

[0060] The area percentage of cholesterol is calculated based on the obtained corrected area value, and the cholesterol content is then determined.

[0061] (Free fatty acids)

[0062] In the egg yolk phospholipid composition of this embodiment, from the viewpoint that the pH decrease can be suppressed and thus an emulsion with better stability can be produced, the content of free fatty acids can be 1.0% by mass or less, preferably 0.8% by mass or less.

[0063] In this invention, free higher fatty acids do not include fatty acids in the form of fatty acid esters that constitute vegetable oils and phospholipids. Higher fatty acids are, for example, straight-chain or branched saturated or unsaturated fatty acids with 6 to 22 carbon atoms (preferably 12 to 20). Examples of higher fatty acids include oleic acid, stearic acid, linoleic acid, palmitic acid, linolenic acid, and myristic acid, with oleic acid being preferred.

[0064] In this invention, the content of free fatty acids is confirmed by the following method: Isopropanol is added to 1.0 g of the sample and dissolved to prepare a 25 mL sample solution. Additionally, palmitic acid is dissolved in heptane to prepare a standard solution containing 1.0 wt% or 0.8 wt% palmitic acid. 1 mL of the prepared sample solution and standard solution is measured into a test tube, and 5.0 mL of a mixture obtained by mixing isopropanol, heptane, and 0.5 mol / L sulfuric acid in a ratio of 40:10:1 is added using a pipette. The mixture is shaken for 1 minute and then allowed to stand for 10 minutes. Then, 3 mL of heptane and 3 mL of distilled water are added to the sample solution, and 2 mL of heptane and 4 mL of distilled water are added to the standard solution. The mixture is sealed and mixed, and allowed to stand for at least 15 minutes to allow for layering. 3 mL of each upper layer is accurately measured into a stoppered test tube, and 1 mL of Nile blue reagent is added. Nitrogen gas is immediately introduced to purge the test tubes, and then titrated with 0.01 mol / L sodium hydroxide solution until the liquid turns purple. The amount of sodium hydroxide solution required to titrate the sample solution is less than the amount required to titrate the standard solution, thus determining the fatty acid content in the sample to be 1.0% by mass or 0.8% by mass.

[0065] From the perspective of better stability, the pH of the egg yolk phospholipid composition of this embodiment is preferably 6.5 or higher and 10 or lower, more preferably 7.0 or higher; on the other hand, it is more preferably 9.5 or lower.

[0066] (Effects)

[0067] Fat emulsions prepared using the egg yolk phospholipid composition of this embodiment have the property of not easily decreasing pH during sterilization. The inventors have discovered that fat emulsions containing egg yolk phospholipid compositions with an LPE content of 1% by mass or less and an amino acid content of 50 mg / 100g or more and 500 mg / 100g or less are less prone to pH decrease during sterilization.

[0068] We hypothesize that the fat emulsion containing the egg yolk phospholipid composition of this embodiment is less prone to pH decrease during sterilization due to the amino acids contained in the egg yolk phospholipid composition. More specifically, we hypothesize that by containing the amount of the amino acid specified above, the amino acid inhibits the pH decrease of the fat emulsion, thereby maintaining the emulsification stability of the fat emulsion.

[0069] 2. Method for manufacturing egg yolk phospholipid composition

[0070] A method for manufacturing the egg yolk phospholipid composition according to one embodiment of the present invention includes the following steps:

[0071] A first egg yolk phospholipid composition (hereinafter also referred to as the "first egg yolk phospholipid composition") having an LPE content of 1% by mass and an amino acid content of 0 mg / 100g or more and 250 mg / 100g or less, and a second egg yolk phospholipid composition (hereinafter also referred to as the "second egg yolk phospholipid composition") having an LPE content of more than 1% by mass and an amino acid content of 250 mg / 100g or more and 700 mg / 100g or less, are mixed to obtain a final egg yolk phospholipid composition ("the egg yolk phospholipid composition of the above embodiment", hereinafter also referred to as the "final egg yolk phospholipid composition") having a lysophosphatidylethanolamine content of 1% by mass and an amino acid content of 50 mg / 100g or more and 500 mg / 100g or less.

[0072] It should be noted that the first egg yolk phospholipid composition can also be mixed with the first raw material egg yolk phospholipid composition described later to obtain the final egg yolk phospholipid composition.

[0073] In addition to the first egg yolk phospholipid composition and the second egg yolk phospholipid composition described above, other components such as separately treated egg yolk phospholipid compositions and amino acids can be further mixed within the scope of not impairing the effects of the present invention to obtain the final phospholipid composition.

[0074] According to the method for manufacturing the egg yolk phospholipid composition of this embodiment, by including a step of mixing the first egg yolk phospholipid composition and the second egg yolk phospholipid composition to obtain the final egg yolk phospholipid composition, a final egg yolk phospholipid composition containing trace amounts (i.e., amino acid content of 50 mg / 100g or more and 500 mg / 100g or less) of amino acids and with LPE content reduced to 1% by mass or less, and with pH not easily decreased during sterilization, can be manufactured in a simple and efficient manner.

[0075] The aforementioned first egg yolk phospholipid composition can be prepared by mixing commercially available amino acids and commercially available egg yolk phospholipids, or it can be prepared by the first resin treatment process described later.

[0076] In order to reduce the content of LPE and amino acids to a specified range, the above-mentioned first egg yolk phospholipid composition is preferably prepared by the first resin treatment process described later.

[0077] In addition, the above-mentioned second egg yolk phospholipid composition can be prepared by mixing commercially available amino acids and commercially available egg yolk phospholipids, or by the second resin treatment process described later, or by mixing the substance prepared by the second resin treatment process described later with commercially available amino acids and / or commercially available egg yolk phospholipids.

[0078] From the perspective of reducing the LPE content and amino acid content to a specified range, the above-mentioned second egg yolk phospholipid composition is preferably prepared by the second resin treatment process described later.

[0079] (An example of a method for manufacturing the egg yolk phospholipid composition of this embodiment using resin treatment)

[0080] An example of the method for manufacturing the egg yolk phospholipid composition of this embodiment includes the following steps: a first resin treatment step (described later), a second resin treatment step (described later), and a step of mixing the first egg yolk phospholipid composition and the second egg yolk phospholipid composition to obtain the final egg yolk phospholipid composition (the egg yolk phospholipid composition of the above embodiment) (hereinafter also simply referred to as the "mixing step").

[0081] The inventors have discovered that fat emulsions using egg yolk phospholipid compositions with reduced LPE through resin treatment tend to have a lower pH during sterilization. This is because amino acids in the egg yolk phospholipid composition are removed along with LPE during resin treatment, and amino acids tend to be preferentially removed compared to LPE during resin treatment.

[0082] In contrast, according to the method for manufacturing the egg yolk phospholipid composition of this example, by including the first resin treatment step, the second resin treatment step, and the step of obtaining the final egg yolk phospholipid composition, a final egg yolk phospholipid composition containing trace amounts of amino acids and with the LPE content reduced to less than 1% by mass, and whose pH does not easily decrease during sterilization, can be manufactured in a simple and efficient manner.

[0083] That is, by using the first resin treatment process described above to obtain a first egg yolk phospholipid composition with a relatively low content of amino acids (e.g., amino acids and LPE), and by using the second resin treatment process described above to obtain a second egg yolk phospholipid composition with a relatively high content of amino acids (e.g., amino acids and LPE), the final egg yolk phospholipid composition can be manufactured by mixing the first egg yolk phospholipid composition with the second egg yolk phospholipid composition in a simple and efficient manner.

[0084] (First resin treatment process)

[0085] The first resin treatment process described above is as follows: a first raw material egg yolk phospholipid composition having an LPE content of more than 1% by mass and less than 3% by mass and an amino acid content of more than 500 mg / 100 g is treated with a first ion exchange resin to obtain the first egg yolk phospholipid composition described above.

[0086] In the first resin treatment step described above and the second resin treatment step described below, there is no particular limitation on the method of resin treatment. For example, it can be carried out by mixing the resin and the solution of the raw material egg yolk phospholipid composition in a solvent, or it can be carried out by filling the resin into a cylindrical container and passing the solution of the raw material egg yolk phospholipid composition through the container.

[0087] The duration of the first resin treatment is, for example, 1 to 30 minutes, preferably more than 1 minute and less than 10 minutes, and this duration is preferably shorter than the duration of the second resin treatment described later. Furthermore, the temperature of the first resin treatment is, for example, performed at 20 to 40°C.

[0088] According to the first resin treatment step described above, since the removal capacity of amino acids (and LPE) is higher than that of the second resin treatment step described later, a first egg yolk phospholipid composition with reduced amino acids (and LPE) can be obtained in a short time.

[0089] (Second resin treatment process)

[0090] The second resin treatment process described above is as follows: the second raw material egg yolk phospholipid composition, which has an LPE content of more than 1% by mass and less than 3% by mass and an amino acid content of more than 500 mg / 100g, is treated with a second ion exchange resin to obtain the second egg yolk phospholipid composition described above.

[0091] The duration of the second resin treatment is, for example, 10 minutes to 1 hour, preferably 20 minutes or more but less than 40 minutes. Furthermore, the temperature of the second resin treatment is, for example, between 20 and 40°C.

[0092] According to the second resin treatment step described above, since the removal capacity of amino acids and LPE is lower than that of the first resin treatment step described above, a second egg yolk phospholipid composition with a higher content of amino acids (and LPE) than the first egg yolk phospholipid composition can be obtained.

[0093] (Raw material: egg yolk phospholipid composition)

[0094] (First and Second) The raw material egg yolk phospholipid composition typically contains 80% to 99% by mass of egg yolk phospholipid (total). For example, in the raw material egg yolk phospholipid composition, the proportion of PC in the egg yolk phospholipid (total) is 60% to 80% by mass, and the proportion of PE is 5% to 20% by mass.

[0095] Furthermore, the LPE content in the raw material egg yolk phospholipid composition is preferably more than 1% by mass; on the other hand, it is preferably less than 3% by mass; in addition, the amino acid content is usually more than 500 mg / 100g. If it is within the above range, the first raw material egg yolk phospholipid composition and the second raw material egg yolk phospholipid composition can be the same raw material, or different raw materials can be used.

[0096] It should be noted that the raw egg yolk phospholipid composition preferably contains at least one selected from serine, threonine, lysine, and arginine, and more preferably contains serine and / or threonine. It should also be noted that the amino acids contained in the egg yolk phospholipid composition of this embodiment can be either originally present in the raw egg yolk phospholipid composition or added to the egg yolk phospholipid composition obtained by treatment with an ion exchange resin; they can be originally present in the raw egg yolk phospholipid composition.

[0097] (Preparation of raw material egg yolk phospholipid composition)

[0098] In the method for manufacturing the egg yolk phospholipid composition of this embodiment, the above-mentioned raw material egg yolk phospholipid composition is obtained by extracting egg yolk with alcohol, removing the alcohol, and then adding acetone at least 18 times the volume of the egg yolk phospholipid and removing the acetone.

[0099] The alcohol used in the alcohol extraction is not particularly limited, and examples include methanol, ethanol, n-propanol, isopropanol, n-butanol, 2-butanol, and tert-butanol. Lower alcohols with 1 or more but 3 or fewer carbon atoms are preferred. An alcohol extract of egg yolk is obtained through this extraction. Furthermore, from the perspective of superior LPE removal efficiency, the alcohol concentration in the extraction solvent is preferably set to more than 95% by mass, and more preferably to more than 96% by mass.

[0100] Furthermore, after removing the alcohol from the alcohol extract of the egg yolk, acetone at least 18 times the volume of the aforementioned raw material egg yolk phospholipid composition is added, and the acetone is removed, thereby obtaining an egg yolk phospholipid composition with reduced cholesterol content. In this case, from the perspective of further reducing cholesterol content, the amount of acetone added is preferably at least 20 times the volume of the raw material egg yolk phospholipid composition.

[0101] (Ion exchange resin)

[0102] Examples of first and second ion exchange resins used in the first and second resin treatment steps described above include strongly acidic or weakly acidic cation exchange resins (Amberlite IR120B, IR124, 200CT, 252, FPC3500, IRC76) and strongly basic or weakly basic anion exchange resins (IRA400J, IRA402BL, IRA404J, IRA458RF, IRA67). These can be used alone or in combination. It is preferable to use cation exchange resins and anion exchange resins in combination, and more preferably, cation exchange resins and anion exchange resins are used in a ratio of more than 1 / 1 (v / v) and less than 1 / 3 (v / v).

[0103] (The rate of decrease in amino acids)

[0104] Compared to the second resin treatment step, the reduction rate of amino acids (and LPEs) in the egg yolk phospholipid composition is higher in the first resin treatment step. Therefore, the first egg yolk phospholipid composition obtained in the first resin treatment step has a lower content of amino acids (e.g., amino acids and LPEs) compared to the second egg yolk phospholipid composition obtained in the second resin treatment step. In other words, the first egg yolk phospholipid composition with a lower content of amino acids (e.g., amino acids and LPEs) compared to the second egg yolk phospholipid composition can be obtained through the first resin treatment step.

[0105] More specifically, in the first resin treatment step described above, the rate of decrease per minute of the amino acid content in the egg yolk phospholipid composition is preferably 100 mg / 100g or more and 250 mg / 100g or less, more preferably 150 mg / 100g or more; on the other hand, it is more preferably 200 mg / 100g or less.

[0106] Furthermore, in the second resin treatment step described above, the rate of decrease per minute of the amino acid content in the egg yolk phospholipid composition is 3 mg / 100g or more and 30 mg / 100g or less, more preferably 5 mg / 100g or more; on the other hand, it is more preferably 25 mg / 100g or less.

[0107] For example, it is preferable to adjust the type and amount of the ion exchange resin in such a way that the rate of reduction of amino acids (and LPE) in the first resin treatment step and the second resin treatment step is within the above range, so as to obtain the first egg yolk phospholipid composition and the second egg yolk phospholipid composition.

[0108] When using ion exchange resin to treat the raw egg yolk phospholipid composition in the first and second resin treatment steps described above, the following methods can be used as examples: The raw egg yolk phospholipid is dissolved in a polar solvent (e.g., alcohols such as methanol and ethanol, water) or a mixture of the above polar solvent and solvents other than the above polar solvent (e.g., halogen solvents such as dichloromethane and chloroform, ketone solvents such as acetone and methyl ethyl ketone, ester solvents such as ethyl acetate, hydrocarbon solvents such as n-hexane and n-pentane, ether solvents such as diethyl ether). After the solution is contacted with the ion exchange resin, the solvent is removed by vacuum distillation. In this case, the above polar solvent and the mixture can be used alone or in combination.

[0109] Ion exchange resins are synthetic resins with a structure in which functional groups such as sulfonic acid groups and quaternary ammonium groups are introduced into a cross-linked three-dimensional polymer matrix. For efficient resin treatment, the resin itself is usually made hydrated with water or similar substances, thereby creating gaps in the structure and increasing the resin's specific surface area before use.

[0110] On the other hand, when treating the raw material egg yolk phospholipid composition with ion exchange resin, the raw material egg yolk phospholipid composition is dissolved in a polar solvent or mixed solvent as described above. Experience shows that when exposed to such polar solvents or mixed solvents, the ion exchange resin will dehydrate over time.

[0111] Once the ion exchange resin is dehydrated, the gaps that existed when it was wetted shrink or close, resulting in a decrease in the ability to remove LPE, especially for components with larger molecular weights like LPE, from effectively contacting the functional groups present in the gaps of the ion exchange resin. Therefore, when using ion exchange resins with larger molecular weights like LPE after dehydration, it is desirable to use ion exchange resins with sufficient gaps, i.e., larger volumes.

[0112] Furthermore, the volume of ion exchange resins is generally influenced by the type of ions paired with functional groups. Specifically, in the case of anion exchange resins, the following tendency is observed: [The text abruptly ends here, so the translation stops as well.] - In comparison, Cl - The more there are, the smaller the volume becomes.

[0113] In view of the above, in the first resin process described above, from the viewpoint that a larger volume ion exchange resin is preferred in order to improve the removal capacity of LPE, the anion exchange resin used in the first resin treatment process described above, which contains Cl- in the anions bound to the functional groups, is preferred. -The ratio is preferably 0.2 equivalent % or less; on the other hand, in the second resin process described above, from the viewpoint of preferably using an ion exchange resin with a smaller volume than that used in the first resin treatment process in order to control the removal efficiency of LPE, the anion exchange resin used in the second resin treatment process is preferably Cl-containing anions bound to functional groups. - The proportion exceeds 0.2 equivalent.

[0114] It should be noted that in the first resin treatment step and the second resin treatment step described above, alumina can also be used instead of the ion exchange resin, or alumina can be used together with the ion exchange resin.

[0115] (Mixed processes)

[0116] The mixing process described above is as follows: the first egg yolk phospholipid composition and the second egg yolk phospholipid composition are mixed to obtain the final egg yolk phospholipid composition (the egg yolk phospholipid composition of the above embodiment).

[0117] In the above mixing process, from the perspective of easily obtaining the egg yolk phospholipid composition (final egg yolk phospholipid composition) of the above embodiment, the ratio of the first egg yolk phospholipid composition to the second egg yolk phospholipid composition (mass ratio) is generally 0.2 or more and 5 or less, preferably 0.5 or more; on the other hand, it is more preferably 4 or less.

[0118] 3. Fat emulsion

[0119] An embodiment of the present invention provides a fat emulsion comprising: emulsified particles containing the egg yolk phospholipid composition of the above embodiment, and an aqueous phase in which the emulsified particles are dispersed.

[0120] The fat emulsion of this embodiment comprises emulsified particles containing the egg yolk phospholipid composition of the above embodiment and an aqueous phase in which the emulsified particles are dispersed, thereby achieving high purity and excellent emulsification stability.

[0121] (Average particle size)

[0122] In the fat emulsion of this embodiment, the average particle size of the emulsion particles is generally 0.1 μm or more and 0.5 μm or less, and from the viewpoint of better stability, it is preferably 0.3 μm or less.

[0123] (oil phase)

[0124] In the fat emulsion of this embodiment, the oil phase constituting the emulsified particles may include, for example, vegetable oils such as soybean oil, sesame oil, rapeseed oil, safflower oil, olive oil, castor oil, corn oil, cottonseed oil, rice bran oil, sunflower seed oil, grapeseed oil, and wheat germ oil, as well as medium-chain triglycerides (MCTs). The vegetable oil is preferably purified vegetable oil.

[0125] (Composition of fat emulsion)

[0126] The fat emulsion in this embodiment is preferably an emulsion in which the aqueous phase is 65% or more and 90% or less by mass and the oil phase is 10% or more and 30% or less by mass.

[0127] (Manufacturing of fat emulsions)

[0128] For example, egg yolk phospholipids and other additives (such as glycerin) are mixed in a specified amount of oil phase using a common homogenizer and homogenized. Then, a necessary amount of water is added and homogenized again using the same homogenizer to transform it into an oil-in-water emulsion (emulsion). This allows the production of the fat emulsion of this embodiment.

[0129] (Lipolysis preparation)

[0130] The lipolysis preparation of the above embodiments is characterized by high purity and excellent emulsification stability by comprising emulsified particles containing the egg yolk phospholipid composition and the agent of the above embodiments, and an aqueous phase in which the emulsified particles are dispersed.

[0131] In the lipolysis emulsion of this embodiment, the average particle size of the emulsified particles is generally 0.1 μm or more and 0.5 μm or less, and from the viewpoint of better stability, it is preferably 0.3 μm or less.

[0132] As the oil phase constituting the lipolysis preparation of this embodiment, for example, the oil phase exemplified as the oil phase constituting the emulsified particles of the above embodiment can be used.

[0133] The agent constituting the hemolytic emulsion of this embodiment is not limited, but lipophilic agents are preferred. Examples of commercially available products include prostaglandins (PGE1), steroidal compounds (dexamethasone palmitate), NSAIDs (flurbiprofen ester), and propofol. More specifically, diazepam, clovidilide butyrate, and indomethacin are also examples.

[0134] The lipolysis preparation of this embodiment is preferably an emulsion in which the aqueous phase is 65% or more and 90% or less by mass, and the oil phase is 10% or more and 30% or less by mass, and the content of the agent is, for example, 5 μg / mL or more and 10% or less by mass.

[0135] (Manufacturing of lipolysis preparations)

[0136] For example, egg yolk phospholipids, pharmaceuticals, and other additives (such as glycerin) are mixed into a specified amount of oil phase using a common homogenizer and homogenized. Then, a necessary amount of water is added, and the homogenizer is used again to homogenize the mixture to transform it into an oil-in-water emulsion (emulsion). This allows the lipolysis preparation of this embodiment to be manufactured.

[0137] Example

[0138] 4. Example

[0139] The present invention will be further described in detail below through embodiments, but the present invention is not limited to the embodiments.

[0140] [Example 1]

[0141] After extraction of dried egg yolks with alcohol (an ethanol-water mixture containing 96% by mass ethanol), the solvent was removed under reduced pressure, and then the mixture was treated with 20 times the amount of acetone to obtain a raw material egg yolk phospholipid composition with the following composition (first raw material egg yolk phospholipid composition / second raw material egg yolk phospholipid composition).

[0142] PC75 quality %

[0143] PE18 quality %

[0144] LPE 2.5% by mass

[0145] CH0.8% by mass

[0146] 0.6% by mass of amino acids

[0147] 100g of the above-mentioned raw material egg yolk phospholipid composition was dissolved in 300ml of solvent (hexane-methanol (80:20), and then added to a container containing 12mL of the first ion exchange resin (product name: Amberlite IR120B (cation exchange resin: H type) and 24mL of Amberlite IRA400 (anion exchange resin: OH type)). The mixture was stirred, and resin treatment was performed (temperature: 20-40°C, time: 4 minutes). This yielded a first egg yolk phospholipid composition with an LPE content of 0.5% by mass and an amino acid content of 10mg / 100g (first resin treatment step). In the first resin treatment step, the amino acid content in the egg yolk phospholipid composition decreased at a rate of 150mg / 100g per minute.

[0148] 100g of the above-mentioned raw material egg yolk phospholipid composition was dissolved in 300ml of solvent (hexane-methanol (80:20), and then added to a container containing 6mL of a second ion exchange resin (product name: Amberlite IR120B (cation exchange resin: H type) and 10mL of Amberlite IRA404 (anion exchange resin: OH type)). The mixture was stirred, and resin treatment was performed (temperature: 20-40°C, time: 20 minutes). This yielded a second egg yolk phospholipid composition with an LPE content of 1.5% by mass and an amino acid content of 400mg / 100g (second resin treatment step). In the above-mentioned second resin treatment step, the amino acid content in the egg yolk phospholipid composition decreased at a rate of 10mg / 100g per minute.

[0149] Then, the mixture was prepared in a ratio of 1:1 (mass ratio) of the first egg yolk phospholipid composition to the second egg yolk phospholipid composition to obtain the final egg yolk phospholipid composition of Example 1 having the following composition. It should be noted that the free fatty acid content in the obtained egg yolk phospholipid composition was 0.2% by mass.

[0150] PC75 quality %

[0151] PE18 quality %

[0152] LPE 1.0 mass %

[0153] CH0.8%

[0154] 0.4% by mass of amino acids

[0155] [Example 2]

[0156] In the second resin treatment step of Example 1 above, 100g of alumina (product name: activated alumina (manufactured by Wako Pure Chemical Industries)) was used instead of the ion exchange resin, and the same treatment as in Example 1 was performed to obtain the egg yolk phospholipid composition of Example 2 having the following composition. It should be noted that the content of free fatty acids in the obtained egg yolk phospholipid composition is 0.2% by mass.

[0157] PC80 quality %

[0158] PE8 quality %

[0159] LPE 0.5% by mass

[0160] CH0.8% by mass

[0161] 0.2% by mass of amino acids

[0162] [Experimental Example 1]

[0163] In Example 1 above, five egg yolk phospholipid compositions with different amino acid contents (test numbers 1-5) were prepared by varying the amount of resin and the reaction time (Table 1). For each phospholipid, a fat emulsion was prepared using a high-pressure homogenizer.

[0164] The pH values ​​of the fat emulsions in Example 1 were 6.5 or higher and 10 or lower, and the average particle size was 0.1 μm or higher and 0.5 μm or lower. It should be noted that in this example, the average particle size was determined using a BECKMAN COULTER particle size distribution measuring device (model: N4 PLUS).

[0165] These samples were adjusted to pH 9.5 with sodium hydroxide aqueous solution, sterilized at 121°C for 20 minutes, and their zeta potential and pH were measured. The results are shown in Table 1.

[0166] It should be noted that in this embodiment, the zeta potential of the fat emulsion was measured using a Malvern Zetasizer Nano Z, and the pH of the fat emulsion was measured using a pH meter (model F-52) manufactured by Horiba Corporation.

[0167]

[0168] As can be understood from Table 1, by using the egg yolk phospholipid composition (test numbers 2-5) of this embodiment, a fat emulsion with minimal pH reduction (specifically, a reduction of less than 2.5) and excellent stability can be obtained.

[0169] In addition, the zeta potential of experiments 2 to 5 remained low, suggesting that the pH decrease during heat sterilization was small.

[0170] Moreover, as can be understood from Example 2, the same effect is obtained when using alumina.

[0171] [Experimental Example 2]

[0172] For the egg yolk phospholipid composition prepared in Test Example 1 above (Test No. 1), after adding serine (Ser), threonine (Thr), valine, leucine, and isoleucine, it was freeze-dried and a fat emulsion (Test Nos. 6-8) was prepared using an ultrasonic homogenizer. These samples were adjusted to pH 9.5 with sodium hydroxide aqueous solution, sterilized at 121°C for 20 minutes, and the pH was measured. The results are shown in Table 2. The pH of each fat emulsion in Test Example 2 was 6.5 or higher and 10 or lower, and the average particle size was 0.1 μm or higher and 0.5 μm or lower.

[0173]

[0174] As can be understood from Table 2, when the total amount of the amino acid is set to 100 parts by mass, a fat emulsion with a total ratio of serine and threonine of 10 parts by mass or more can be easily obtained with less pH reduction and excellent stability after heat sterilization.

[0175] [Experimental Example 3]

[0176] For Sample 1 prepared in Example 1 above, after adding serine (Ser), threonine (Thr), arginine (Arg), lysine (Lys), valine, leucine, and isoleucine, it was freeze-dried and a fat emulsion was prepared using an ultrasonic homogenizer. These samples were adjusted to pH 9.5 with sodium hydroxide aqueous solution, sterilized at 121°C for 20 minutes, and the pH was measured. The results are shown in Table 3. The pH of each fat emulsion in Example 3 was 6.5 or higher and 10 or lower, and the average particle size was 0.1 μm or higher and 0.5 μm or lower.

[0177]

[0178] As can be understood from Table 3, when the total amount of the amino acid is set to 100 parts by mass, the combined proportion of serine, threonine, arginine and lysine in the amino acid is more than 10 parts by mass, and a fat emulsion with less pH reduction and excellent stability after heat sterilization can be easily obtained.

[0179] [Experimental Example 4]

[0180] The egg yolk phospholipid composition of Example 1 (6g) and soybean oil (100g) were mixed and dispersed at high speed. After adding 384g of distilled water and 10g of glycerol midway through the process, coarse emulsification was performed (stirring speed: 10000rpm, stirring time: 40 minutes). Then, fine emulsification (high-pressure emulsification: 500kg / cm³) was carried out. 2 The solution was repeatedly passed through the container 10-20 times. Then, after pH adjustment and filtration, it was filled into a glass vial to obtain the fat emulsion (O / W emulsion containing 20% ​​soybean oil by mass) of Experimental Example 4.

[0181] After sterilizing the obtained fat emulsion at 121°C for 20 minutes, the bottle was opened and the average particle size and pH of the fat emulsion were measured. The results showed that the fat emulsion in Experiment 4 had little change in average particle size before and after sterilization, and the decrease in pH after sterilization was suppressed, indicating high stability.

[0182] [Experimental Example 5]

[0183] 6g of the egg yolk phospholipid composition from Example 1, 50g of soybean oil, and 10g of the reagent (indomethacin) were stirred and dispersed at high speed. After adding 424g of distilled water and 10g of glycerin midway through the process, coarse emulsification was performed (stirring speed: 10000 rpm, stirring time: 40 minutes), followed by fine emulsification (high-pressure emulsification: 500 kg / cm³). 2 The solution was repeatedly passed through the container 10–20 times. Then, after pH adjustment and filtration, it was filled into a glass vial to obtain the lipolysis preparation of Example 5 (an O / W emulsion containing 10% by mass soybean oil).

[0184] After sterilizing the obtained lipolysis preparation at 121°C for 20 minutes, the bottle was opened and the average particle size and pH of the lipolysis preparation were measured. The results showed that the lipolysis preparation of Experiment Example 5 had little change in average particle size before and after sterilization, and the decrease in pH after sterilization was inhibited, indicating high stability.

Claims

1. A lecithin composition containing lysophosphatidylethanolamine at a content of less than 1% by mass and amino acids at a content of more than 50 mg / 100g and less than 500 mg / 100g, wherein, When the total amount of the amino acid is set to 100 parts by mass, the combined proportion of serine and threonine in the amino acid is 10 parts by mass or more, and the pH of the egg yolk phospholipid composition is 6.5 or higher and 10 or lower. When the total amount of the amino acid is set to 100 parts by mass, the combined proportion of serine, threonine, lysine and arginine in the amino acid is more than 15 parts by mass. Wherein, when the total amount of egg yolk phospholipids is set to 100 parts by mass, the content of the amino acids is more than 0.06 parts by mass and less than 0.6 parts by mass relative to the content of egg yolk phospholipids; The egg yolk phospholipid composition contains phosphatidylcholine, wherein when the content of the phosphatidylcholine is set to 100 parts by mass, the content of the amino acid is more than 0.1 parts by mass and less than 0.8 parts by mass relative to the content of the phosphatidylcholine. The egg yolk phospholipid composition contains phosphatidylethanolamine. When the content of the phosphatidylethanolamine is set to 100 parts by weight, the content of the amino acid is 0.2 parts by weight or more and 10 parts by weight or less relative to the content of the phosphatidylethanolamine, and the content of the hemolysin phosphatidylethanolamine is 3 parts by weight or more and 15 parts by weight or less relative to the content of the phosphatidylethanolamine. The egg yolk phospholipid composition contains 65% or more of phosphatidylcholine and 7% or more of phosphatidylethanolamine.

2. The egg yolk phospholipid composition according to claim 1, wherein, The cholesterol content is less than 1.5% by mass.

3. The egg yolk phospholipid composition according to claim 1, wherein, The content of free fatty acids is less than 1.0% by mass.

4. A fat emulsion comprising: Emulsified particles containing the egg yolk phospholipid composition according to any one of claims 1 to 3, and The aqueous phase in which the emulsion particles are dispersed.

5. A lipolysis preparation comprising: Emulsified particles containing the egg yolk phospholipid composition and pharmaceutical agent as described in any one of claims 1 to 3, and The aqueous phase in which the emulsion particles are dispersed.

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