Probiotic-containing self-emulsifying preparation

Abstract

The present invention provides a probiotic-containing self-emulsifying composition that retains probiotics at a high survival rate and exhibits excellent emulsion / dispersion properties when mixed with liquid food / beverage or a cosmetic.　Provided is a lactic-acid-bacteria-containing self-emulsifying composition obtained by mixing probiotics, an oil and fat, and a specific composite emulsifier.

Description

Self-emulsifying formulation containing probiotics

[0001] The present invention relates to a self-emulsifying composition containing probiotics, which can emulsify and disperse probiotics in a liquid food, drink, or cosmetic product by mixing with the liquid food, drink, or cosmetic product.

[0002] Probiotics are live microorganisms that have a beneficial effect on the host when ingested in sufficient amounts. Lactic acid bacteria, a typical example of probiotics, are microorganisms present in the intestines of humans and animals. They have traditionally been known to promote intestinal peristalsis and regulate the intestines, but recent research has also revealed that they may also improve immune function, reduce blood triglyceride and cholesterol levels, and improve skin function.

[0003] In response to this situation, interest is growing in foods and beverages that allow people to actively consume probiotics such as lactic acid bacteria, especially in drink-type products such as probiotic drinks and drinkable yogurt, due to their ease of intake.In addition, because the effects of resident skin bacteria are expected to improve skin moisture and have whitening effects, their application in cosmetics is also being considered.

[0004] A known method for dispersing probiotics in liquid foods and beverages involves mixing an aqueous phase containing probiotics, oils and fats, and a lipophilic emulsifier to prepare a W / O type emulsion, and then mixing the W / O type emulsion with an aqueous phase containing a hydrophilic emulsifier to form a W / O / W type emulsion (Patent Document 1).

[0005] Japanese Patent Application Laid-Open No. 2007-60919

[0006] The inventors have conceived the idea of ​​storing and / or carrying a composition containing probiotics, oils and fats, and an emulsifier without immediately mixing it with an aqueous phase, and then mixing it with a liquid food, drink, or cosmetic of one's choice when needed to prepare a probiotic-containing food, drink, or cosmetic. This method of use not only significantly increases the flexibility of opportunities to ingest probiotics from food and drink, but also makes it easier for the probiotics to penetrate the skin and realize their effects, and allows one to select a liquid food, drink, or cosmetic in which to disperse the probiotics according to one's preferences.

[0007] In order to realize the above-mentioned usage of probiotics, the storage stability of the probiotics in the composition state prior to mixing with liquid food and beverage products or cosmetics is important.

[0008] The inventors of the present invention have preliminarily confirmed the survival rate of probiotics in a mixture of probiotics, oils and fats, and an emulsifier, and have found that the results are not good and there is room for improvement.

[0009] Some emulsifiers, such as polysorbate 80 (Tween 80), have a relatively small adverse effect on the viability of probiotics; however, these emulsifiers require a high emulsifier ratio in order to be ultimately mixed and emulsified with liquid foods, beverages, or cosmetics, and there is room for improvement in terms of consumer image, mouthfeel, taste, and usability when mixed with liquid foods, beverages, or cosmetics.

[0010] In view of the above circumstances, an object of the present invention is to provide a composition containing probiotics, oils and fats, and an emulsifier, in which the survival rate of the probiotics is improved, and in particular, to provide a composition in which the proportion of the emulsifier in the composition can be reduced.

[0011] As a result of intensive research into the above-mentioned problems, the present inventors have found that the use of a composite emulsifier consisting of at least two different emulsifiers can provide stable emulsifying power and improve the survival rate of probiotics.

[0012] That is, the present invention encompasses the following aspects. [1] A composition containing probiotics, an emulsifier, and an oil or fat, wherein the emulsifier is 1) a complex emulsifier containing monooleic acid diglyceride and polysorbate, or 2) a complex emulsifier containing monooleic acid diglyceride and polyglycerol laurate. [2] The composition of [1], wherein the probiotic is a lactic acid bacterium. [3] The composition of [1] or [2], wherein the complex emulsifier has a weight ratio of monooleic acid diglyceride to polysorbate of 2:1 to 1:2. [4] The composition of [1] or [2], wherein the complex emulsifier has a weight ratio of monooleic acid diglyceride to polyglycerol laurate of 2:1 to 1:2. [5] Any one of the compositions of [1] to [4], wherein the weight ratio of the oil or fat to the complex emulsifier is 7:3 to 2:3. [6] Any one of the compositions of [1] to [5], wherein the polysorbate is polysorbate 80. [7] Any one of the compositions of [1] to [6], wherein the polyglycerol laurate is an ester of polyglycerol 10 and lauric acid. [8] Any one of the compositions of [1] to [7], wherein the oil or fat is MCT (medium-chain triglyceride). [9] Any one of the compositions of [1] to [8], which is used as an additive for food, beverage, or cosmetics.

[10] Any one of the compositions of [1] to [8], which is a probiotic-containing food, beverage, or probiotic-containing cosmetic.

[0013] This specification includes the disclosure of Japanese Patent Application No. 2024-052895, from which the present application claims priority.

[0014] According to the present invention, the survival rate of probiotics can be improved in a composition (probiotic-containing self-emulsifying composition) that contains probiotics, oils and fats, and an emulsifier and can be mixed with a liquid food or beverage or cosmetic to prepare a probiotic-containing food or beverage or a probiotic-containing cosmetic.

[0015] Furthermore, the self-emulsifying composition can be stored and / or carried, and when needed, can be mixed with liquid food and beverage products or cosmetics to prepare probiotic-containing food and beverage products or probiotic-containing cosmetics, greatly improving the convenience of taking probiotics.

[0016] The present invention will be described in detail below.

[0017] The present invention relates to a composition containing probiotics, an oil or fat, and an emulsifier. Since the composition of the present invention can be easily emulsified when mixed with a liquid food, drink, or cosmetic product, thereby dispersing the probiotics in the food, drink, or cosmetic product, it is also referred to herein as a probiotic-containing self-emulsifying composition, or simply as a self-emulsifying composition.

[0018] As used herein, "probiotics" refers to live microorganisms that, when ingested in sufficient amounts, confer a beneficial effect on the host. The probiotics contained in the probiotic-containing self-emulsifying composition of the present invention are not particularly limited as long as they can be ingested and used as a food, health food, or cosmetic. Examples of probiotics contained in the probiotic-containing self-emulsifying composition of the present invention include normal inhabitant bacteria in the body, such as lactic acid bacteria, bifidobacteria, butyric acid bacteria, Bacillus subtilis, and Akkermansia. Examples of the probiotics contained in the probiotic-containing self-emulsifying composition of the present invention include bacteria of the genus Lactiplantibacillus, bacteria of the genus Lactobacillus, bacteria of the genus Lacticaseibacillus, bacteria of the genus Latilactobacillus, bacteria of the genus Lentilactobacillus, bacteria of the genus Levilactobacillus, bacteria of the genus Ligilactobacillus, and the like. Examples of the probiotics contained in the probiotic-containing self-emulsifying composition of the present invention include bacteria of the genus Limosilactobacillus, bacteria of the genus Loigolactobacillus, bacteria of the genus Lopidilactobacillus, bacteria of the genus Lysinibacillus, bacteria of the genus Pediococcus, bacteria of the genus Streptococcus, bacteria of the genus Lactococcus, bacteria of the genus Bifidobacterium, bacteria of the genus Enterococcus, and bacteria of the genus Clostridium. The probiotics contained in the probiotic-containing self-emulsifying composition of the present invention may preferably be lactic acid bacteria. The lactic acid bacteria contained in the probiotic-containing self-emulsifying composition of the present invention are not particularly limited as long as they are lactic acid bacteria that can be ingested and used as foods, health foods, or cosmetics, and examples thereof include lactic acid bacteria belonging to the genera Lactobacillus, Pediococcus, Streptococcus, Lactococcus, etc. Among these, lactic acid bacteria belonging to the genus Lactobacillus are preferred, and Lactobacillus plantarum is more preferred.

[0019] The oils and fats contained in the probiotic-containing self-emulsifying composition of the present invention are not particularly limited as long as they are ingestible and usable as foods or cosmetics, and examples thereof include salad oil, olive oil, palm oil, sunflower oil, soybean oil, rapeseed oil, avocado oil, camellia oil, turtle oil, macadamia nut oil, corn oil, mink oil, egg yolk oil, sesame oil, persic oil, wheat germ oil, sasanqua oil, castor oil, linseed oil, safflower oil, cottonseed oil, perilla oil, peanut oil, tea seed oil, kaya oil, rice bran oil, Chinese tung oil, Japanese tung oil, jojoba oil, rice germ oil, cacao butter, coconut oil, horse tallow, hydrogenated coconut oil, palm oil, beef tallow, palm kernel oil, hydrogenated oil, Japan wax, hydrogenated castor oil, shea butter, beeswax, carnauba wax, candelilla wax, Ibotaro wax, and MCT (medium-chain triglycerides). Among these, palm oil and sunflower oil are preferred from the viewpoint of oxidation stability, and MCT (medium chain fatty acid triglyceride) is more preferred.

[0020] The probiotic-containing self-emulsifying composition of the present invention uses a composite emulsifier containing or consisting of diglyceride monooleate and other emulsifiers, such as polysorbate (polyoxyethylene sorbitan fatty acid ester) and / or polyglycerin laurate.

[0021] Examples of polyoxyethylene sorbitan fatty acid esters include polysorbate 80 (polyoxyethylene sorbitan monooleate), polysorbate 60 (polyoxyethylene sorbitan monostearate), and polysorbate 20 (polyoxyethylene sorbitan monolaurate), with polysorbate 80 being preferred.

[0022] Examples of polyglycerol laurate esters include esters of lauric acid and polyglycerol having a degree of polymerization of 2 to 10, with esters of lauric acid and polyglycerol having a degree of polymerization of 6 to 10 being preferred, and esters of lauric acid and polyglycerol having a degree of polymerization of 10 (polyglycerol 10) being more preferred.

[0023] The composite emulsifier used in the present invention can be easily prepared by mixing and stirring monooleic acid diglyceride with the other emulsifiers described above.

[0024] The weight ratio of the monooleic acid diglyceride to the other emulsifiers in the composite emulsifier is, for example, 2:1 to 1:2, preferably 1.5:1 to 1:1.5, more preferably 1.2:1, and even more preferably 1:1.

[0025] The probiotic-containing self-emulsifying composition may contain emulsifiers other than monooleic acid diglyceride, polysorbate, and polyglycerol laurate, as long as they do not adversely affect the survival rate of the probiotics in the probiotic-containing self-emulsifying composition.

[0026] The probiotic-containing self-emulsifying composition of the present invention can be prepared by mixing and stirring the probiotics, oils and fats, and composite emulsifier.

[0027] Probiotics can be used in the preparation of a probiotics-containing self-emulsifying composition in the form of a freeze-dried powder or wet cells obtained by centrifuging a culture solution and washing it as necessary. The number of viable probiotics in the freeze-dried powder or wet cells can be adjusted appropriately depending on the specifications of the probiotics-containing self-emulsifying composition to be prepared.

[0028] From the viewpoint of the survival rate of the probiotics, it is preferable that the water content in the probiotic-containing self-emulsifying composition is low, and it is preferable to use probiotics in the form of a freeze-dried powder.

[0029] The number of live probiotic bacteria in the probiotic-containing self-emulsifying composition is not particularly limited, but is preferably 1.0 x 10 ７ ~5.0 x 10 １２ cfu / g, 1.0×10 ７ ~1.0 x 10 １２ cfu / g, 1.0×10 ８ ~1.0 x 10 １２ cfu / g, 1.0×10 ９ ~1.0 x 10 １２ cfu / g, 1.0×10 １０ ~1.0 x 10 １２ cfu / g, 1.0×10１１ ~1.0 x 10 １２ cfu / g, 3.0×10 １１ ~7.0 x 10 １１ cfu / g, 4.0×10 １１ ~6.5 x 10 １１ cfu / g or 2.0 x 10 １１ ~4.5 x 10 １１ An example is cfu / g.

[0030] The content of the complex emulsifier in the probiotic-containing self-emulsifying composition is not particularly limited, and may be, for example, 10 to 60% by weight, 20 to 60% by weight, 30 to 60% by weight, 30 to 55% by weight, or 30 to 50% by weight. The content of the oil or fat in the probiotic-containing self-emulsifying composition is not particularly limited, and may be, for example, 5 to 80% by weight, 10 to 90% by weight, 20 to 80% by weight, 30 to 75% by weight, or 40 to 70% by weight.

[0031] The order of adding and mixing the probiotics, fats and oils, and composite emulsifier is not particularly limited, but it is preferable to mix the fats and oils with the composite emulsifier in advance, and then mix the mixture with the probiotics.

[0032] The weight ratio of the fat and oil to the complex emulsifier is, for example, 7:3 to 2:3, and preferably 3:2 to 1:1.

[0033] The temperature during mixing and stirring should not be too high from the viewpoint of the survival rate of the probiotics, and is preferably, for example, 20°C to 35°C. Depending on the viscosity of the oils and / or emulsifiers used, it may be preferable to heat the oils and / or emulsifiers in advance to increase their fluidity before mixing. The degree of heating is within a range that does not result in an excessively high temperature during mixing, for example, 25 to 45°C, preferably 30 to 40°C. The mixing and stirring operation can be carried out using general-purpose equipment and devices normally used for this purpose.

[0034] The probiotics-containing composition of the present invention is prepared in the form of a highly viscous liquid through the above-mentioned procedures.

[0035] The prepared probiotics-containing self-emulsifying composition is preferably sealed and stored under conditions / environments of 4 to 40°C, 4 to 35°C, 4 to 30°C, or 4 to 25°C until it is mixed with a liquid food or beverage or a cosmetic product and used.

[0036] When added to an aqueous liquid, the probiotic-containing self-emulsifying composition of the present invention emulsifies to form an O / W emulsion, thereby dispersing the probiotics in the aqueous composition. The probiotic-containing self-emulsifying composition of the present invention has improved probiotic survival. The probiotic-containing self-emulsifying composition of the present invention also has excellent water dispersibility. Water dispersibility can be evaluated, for example, as described in the Examples below. Specifically, for example, 1 g of the probiotic-containing self-emulsifying composition of the present invention is suspended in 30 mL of water and allowed to stand. In this case, a composition identical to the probiotic-containing self-emulsifying composition of the present invention except that it does not contain probiotics may be used instead of the probiotic-containing self-emulsifying composition of the present invention. If no separation occurs after 3 hours of standing, the probiotic-containing self-emulsifying composition of the present invention can be determined to have excellent water dispersibility.

[0037] The probiotic-containing self-emulsifying composition of the present invention can be used, for example, for addition to an aqueous liquid. The aqueous liquid is not particularly limited as long as it contains water, and can be, for example, a food or drink (e.g., water, tea, juice, or other beverages, liquid diets, smoothies, yogurt, protein, etc.) or a cosmetic (lotion, emulsion, cleanser, cream, mist, balm, etc.). The food or drink is preferably a liquid food or drink, and the cosmetic is preferably a liquid cosmetic. The probiotic-containing self-emulsifying composition of the present invention can be, for example, a composition for dispersing probiotics in an aqueous liquid by adding it to the aqueous liquid and emulsifying it. Furthermore, the probiotic-containing self-emulsifying composition of the present invention can be ingested as a food or drink itself or used as a cosmetic.

[0038] The probiotic-containing self-emulsifying composition of the present invention can be mixed with beverages such as water, tea, and juice, as well as liquid foods and smoothies, to easily prepare probiotic-containing foods and beverages when needed. Furthermore, the probiotic-containing self-emulsifying composition of the present invention can be mixed with lotions, emulsions, cleansers, creams, mists, balms, and the like to easily prepare probiotic-containing cosmetics when needed. While the specific preparation method is not particularly limited, examples of the preparation method include mixing 1 to 2 drops of the probiotic-containing self-emulsifying composition with a glass of water, mixing 5 to 10 drops (200 to 500 mg) of the probiotic-containing self-emulsifying composition with a protein shaker (350 to 1,000 mL), mixing 0.5 to 1.0 mL of the lactic acid bacteria-containing self-emulsifying composition with a smoothie (200 to 500 mL), and mixing 1 to 2 drops of the probiotic-containing self-emulsifying composition with a portion of lotion dispensed into the palm of the hand.

[0039] The amount to be added to liquid food, drink, or cosmetics can be adjusted appropriately depending on the type of food, drink, or cosmetic product and the amount of probiotics desired to be ingested, and is not particularly limited; however, examples of the amount of the probiotic-containing self-emulsifying composition to be added per 30 g of liquid food, drink, or cosmetic product include about 0.001 to 5 g, 0.005 to 4.5 g, 0.01 to 4 g, 0.05 to 3.5 g, 0.1 to 3 g, 0.5 to 2 g, or 0.1 to 1 g.

[0040] The present invention also provides a method for producing the probiotic-containing self-emulsifying composition of the present invention. The method includes, for example, mixing and stirring the probiotic, fat, and composite emulsifier. In the method, the types, amounts, and mixing and stirring methods of the probiotic, composite emulsifier, and fat are as described above.

[0041] The present invention also provides probiotic-containing foods and beverages or probiotic-containing cosmetics. The probiotic-containing foods and beverages or probiotic-containing cosmetics contain probiotics, a complex emulsifier, and fats and oils, with the types and amounts of the probiotics, complex emulsifier, and fats and oils being as described above. Examples of probiotic-containing foods and beverages include beverages such as water, tea, and juice, as well as liquid foods, smoothies, yogurt, and protein foods. Examples of probiotic-containing cosmetics include lotions, emulsions, cleansers, creams, mists, balms, and the like.

[0042] The present invention also provides a method for improving the viability of probiotics in a composition containing probiotics, an emulsifier, and an oil or fat, the method comprising preparing the composition by mixing the probiotics, the complex emulsifier, and the oil or fat. In this method, the types of probiotics, the complex emulsifier, and the oil or fat, the mixing amounts, and the mixing and stirring method are as described above. As used herein, "improving the viability of probiotics in a composition" can mean, for example, improving the viability of probiotics (e.g., improving by 5% or more) compared to an identical composition except that monooleic acid diglyceride alone is used as the emulsifier. The viability of probiotics can be evaluated, for example, using the viability of probiotics after a predetermined period of storage of the composition (i.e., the ratio of the viable cell count of probiotics in the composition after storage to the viable cell count of probiotics in the composition at the start of storage) as an indicator. Improving the viability of probiotics can mean improving the viability of probiotics when the composition is stored, for example, at 4 to 40°C, 4 to 35°C, 4 to 30°C, or 4 to 25°C. Increasing the viability of the probiotics can also be, for example, increasing the viability of the probiotics when the composition is stored for 5 days to 1 year, 10 days to 6 months, 15 days to 2 months, or 1 month.

[0043] The present invention will be explained in more detail below with reference to examples.

[0044] (Example 1) (Measurement of Viability of Lactic Acid Bacteria) A lyophilized powder of Lactobacillus plantarum KABP-023 strain (CECT 7485) was used as lactic acid bacteria, MCT was used as fat and oil, and a composite emulsifier (weight ratio: DO-100V:Tween80=1:1) consisting of diglyceride monooleate (DO-100V, manufactured by Riken Vitamin) and polysorbate 80 (Tween80, manufactured by Kanto Chemical) was mixed to prepare a self-emulsifying composition containing lactic acid bacteria (initial cell count: 6.0 × 10 １１ cfu / g, emulsifier ratio in the composition 50.0 wt%, emulsifier content in the composition 47.6 wt%, oil / fat content in the composition 47.6 wt%). Note that the "emulsifier ratio in the composition" refers to the ratio of the amount of emulsifier to the total amount of emulsifier and oil / fat in the composition.

[0045] The obtained self-emulsifying composition containing lactic acid bacteria was sealed in a centrifuge tube (manufactured by IWAKI, 15 mL volume), and the centrifuge tube was then placed in an aluminum pouch and sealed. After storage at 25°C for 30 days, the viability of the lactic acid bacteria (the ratio of the number of live bacteria after storage to the initial number of bacteria) was measured and found to be 61%.

[0046] (Evaluation of dispersibility) Except for not mixing lactic acid bacteria, fats and oils and emulsifiers were mixed in the same manner as the lactic acid bacteria-containing self-emulsifying composition used in measuring survival rate, to prepare a composition for dispersibility evaluation (emulsifier ratio in composition 50.0 wt%, emulsifier content in composition 50.0 wt%, fats and oils content in composition 50.0 wt%). 1 g of the prepared composition was suspended in 30 mL of water and allowed to stand, and the dispersibility in water was confirmed, and no separation occurred even after 3 hours of standing.

[0047] (Example 2) (Measurement of Viability of Lactic Acid Bacteria) A self-emulsifying composition containing lactic acid bacteria was prepared in the same manner as in Example 1, using a composite emulsifier (weight ratio: DO-100V:ML-750 = 1:1) consisting of monooleic acid diglyceride (DO-100V, manufactured by Riken Vitamin) and polyglycerin 10-laurate (SY Glystar ML-750, manufactured by Sakamoto Pharmaceutical Co., Ltd.) as an emulsifier (initial cell count: 6.0 × 10 １１ cfu / g, emulsifier ratio in the composition 50.0% by weight, emulsifier content in the composition 47.6% by weight, oil and fat content in the composition 47.6% by weight.

[0048] The obtained self-emulsifying composition containing lactic acid bacteria was stored at 25°C for 30 days in the same manner as in Example 1, and the survival rate of the lactic acid bacteria was measured and found to be 64%.

[0049] (Evaluation of dispersibility) Except for not mixing lactic acid bacteria, fats and oils and emulsifiers were mixed in the same manner as the lactic acid bacteria-containing self-emulsifying composition used in measuring survival rate, to prepare a composition for dispersibility evaluation (emulsifier ratio in composition 50.0 wt%, emulsifier content in composition 50.0 wt%, fats and oils content in composition 50.0 wt%). 1 g of the prepared composition was suspended in 30 mL of water and allowed to stand, and the dispersibility in water was confirmed, and no separation occurred even after 3 hours of standing.

[0050] Comparative Example 1 A lactic acid bacteria-containing self-emulsifying composition was prepared by mixing monooleic acid diglyceride (DO-100V, manufactured by Riken Vitamin) as an emulsifier in the same manner as in Example 1 (initial cell count: 4.5 × 10 １１ cfu / g, emulsifier ratio in the composition 50.0% by weight, emulsifier content in the composition 47.6% by weight, oil and fat content in the composition 47.6% by weight.

[0051] The obtained self-emulsifying composition containing lactic acid bacteria was stored at 25°C for 30 days in the same manner as in Example 1, and the survival rate of the lactic acid bacteria was measured and found to be 50%.

[0052] Comparative Example 2 A lactic acid bacteria-containing self-emulsifying composition was prepared in the same manner as in Example 1, except that polysorbate 80 (Tween 80, manufactured by Kanto Chemical) was used as an emulsifier (initial cell count: 4.5 × 10 １１ cfu / g, emulsifier ratio in the composition 66.7% by weight, emulsifier content in the composition 63.5% by weight, oil and fat content in the composition 31.7% by weight.

[0053] The obtained self-emulsifying composition containing lactic acid bacteria was stored at 25°C for 30 days in the same manner as in Example 1, and the survival rate of the lactic acid bacteria was measured and found to be 60%.

[0054] (Comparative Example 3) A self-emulsifying composition containing lactic acid bacteria was prepared in the same manner as in Comparative Example 1, except that polyglycerin 10-laurate (SY Glystar ML-750, manufactured by Sakamoto Pharmaceutical Co., Ltd.) was used as an emulsifier (initial number of bacteria: 4.5 × 10 １１cfu / g, emulsifier ratio in the composition 85.7 wt%, emulsifier content in the composition 81.6 wt%, fat / oil content in the composition 13.6 wt%). The obtained lactic acid bacteria-containing self-emulsifying composition was stored at 25°C for 30 days in the same manner as in Example 1, and the survival rate of the lactic acid bacteria was measured and found to be 45%.

[0055] (Comparative Example 4) (Measurement of Viability of Lactic Acid Bacteria) A self-emulsifying composition containing lactic acid bacteria was prepared in the same manner as in Example 1, using a composite emulsifier (weight ratio: DO-100V:MM-750 = 1:1) consisting of monooleic acid diglyceride (DO-100V, manufactured by Riken Vitamin) and polyglycerin 10-myristyl phosphate (SY Glystar MM-750, manufactured by Sakamoto Pharmaceutical Co., Ltd.) (initial cell count: 6.0 × 10 １１ cfu / g, emulsifier ratio in the composition 50.0% by weight, emulsifier content in the composition 47.6% by weight, oil and fat content in the composition 47.6% by weight.

[0056] The obtained self-emulsifying composition containing lactic acid bacteria was stored at 25°C for 30 days in the same manner as in Example 1, and the survival rate of the lactic acid bacteria was measured and found to be 45%.

[0057] (Evaluation of dispersibility) Except for not mixing lactic acid bacteria, fats and oils and emulsifiers were mixed in the same manner as the lactic acid bacteria-containing self-emulsifying composition used in measuring survival rate, to prepare a composition for dispersibility evaluation (emulsifier ratio in composition 50.0 wt%, emulsifier content in composition 50.0 wt%, fats and oils content in composition 50.0 wt%). 1 g of the prepared composition was suspended in 30 mL of water and allowed to stand, and the dispersibility in water was confirmed, and no separation occurred even after 3 hours of standing.

[0058] (Comparative Example 5) (Measurement of Viability of Lactic Acid Bacteria) A self-emulsifying composition containing lactic acid bacteria was prepared in the same manner as in Example 1, using monooleic acid diglyceride (DO-100V, manufactured by Riken Vitamin) as an emulsifier (initial number of bacteria: 4.5 × 10 １１ cfu / g, emulsifier ratio in the composition 50.0% by weight, emulsifier content in the composition 47.6% by weight, oil and fat content in the composition 47.6% by weight.

[0059] The obtained self-emulsifying composition containing lactic acid bacteria was stored at 25°C for 20 days in the same manner as in Example 1, and the survival rate of the lactic acid bacteria was measured and found to be 59%.

[0060] (Evaluation of Dispersibility) A composition for dispersibility evaluation was prepared by mixing fats and oils and an emulsifier in the same manner as the lactic acid bacteria-containing self-emulsifying composition used in measuring the survival rate, except that lactic acid bacteria were not mixed in (emulsifier ratio in the composition: 50.0 wt%, emulsifier content in the composition: 50.0 wt%, fats and oils content in the composition: 50.0 wt%). 1 g of the prepared composition was suspended in 30 mL of water and allowed to stand to check its dispersibility in water. Separation occurred within 1 to 3 hours after standing.

[0061] (Comparative Example 6) A composition for evaluating dispersibility was prepared in the same manner as in Example 1, except that polysorbate 80 (Tween 80, manufactured by Kanto Chemical) was used as the emulsifier (emulsifier ratio in the composition: 50.0 wt %, oil ratio in the composition: 50.0 wt %). Because the obtained composition separated into two layers, dispersibility could not be evaluated, and it was unsuitable as a lactic acid bacteria-containing self-emulsifying composition.

[0062] (Comparative Example 7) (Measurement of Viability of Lactic Acid Bacteria) A self-emulsifying composition containing lactic acid bacteria was prepared in the same manner as in Example 1, using polyglycerol 10-laurate (SY Glystar ML-750, manufactured by Sakamoto Pharmaceutical Co., Ltd.) as an emulsifier (initial number of bacteria: 4.5 × 10 １１ cfu / g, emulsifier ratio in the composition 85.7% by weight, emulsifier content in the composition 81.6% by weight, oil and fat content in the composition 13.6% by weight.

[0063] The obtained self-emulsifying composition containing lactic acid bacteria was stored at 25°C for 20 days in the same manner as in Example 1, and the survival rate of the lactic acid bacteria was measured and found to be 60%.

[0064] (Evaluation of dispersibility) Except for not mixing lactic acid bacteria, fats and oils and emulsifiers were mixed in the same manner as the lactic acid bacteria-containing self-emulsifying composition used in measuring survival rate, to prepare a composition for dispersibility evaluation (emulsifier ratio in composition 85.7 wt%, emulsifier content in composition 85.7 wt%, fats and oils content in composition 14.3 wt%). 1 g of the prepared composition was suspended in 30 mL of water and allowed to stand, and the dispersibility in water was confirmed, and no separation occurred even after 3 hours of standing.

[0065] (Comparative Example 8) (Measurement of Viability of Lactic Acid Bacteria) A self-emulsifying composition containing lactic acid bacteria was prepared in the same manner as in Example 1, using polyglycerol 10-myristyl phosphate (SY Glystar MM-750, manufactured by Sakamoto Pharmaceutical Co., Ltd.) as an emulsifier (initial number of bacteria: 4.5 × 10 １１ cfu / g, emulsifier ratio in the composition 88.9% by weight, emulsifier content in the composition 84.7% by weight, oil and fat content in the composition 10.6% by weight.

[0066] The obtained self-emulsifying composition containing lactic acid bacteria was stored at 25°C for 20 days in the same manner as in Example 1, and the survival rate of the lactic acid bacteria was measured and found to be 57%.

[0067] (Evaluation of dispersibility) Except for not mixing lactic acid bacteria, fats and oils and emulsifiers were mixed in the same manner as the lactic acid bacteria-containing self-emulsifying composition used in measuring survival rate, to prepare a composition for dispersibility evaluation (emulsifier ratio in composition: 88.9 wt%, emulsifier content in composition: 88.9 wt%, fats and oils content in composition: 11.1 wt%). 1 g of the prepared composition was suspended in 30 mL of water and allowed to stand, and the dispersibility in water was confirmed, and no separation occurred even after 3 hours of standing.

[0068] (Comparative Example 9) (Measurement of Viability of Lactic Acid Bacteria) A self-emulsifying composition containing lactic acid bacteria was prepared in the same manner as in Example 1, using polyglycerin 6-oleate (SY Glystar MO-5S, manufactured by Sakamoto Pharmaceutical Co., Ltd.) as an emulsifier (initial number of bacteria: 4.5 × 10 １１ cfu / g, emulsifier ratio in the composition 66.7% by weight, emulsifier content in the composition 63.5% by weight, oil and fat content in the composition 31.7% by weight.

[0069] The obtained self-emulsifying composition containing lactic acid bacteria was stored at 25°C for 20 days in the same manner as in Example 1, and the survival rate of the lactic acid bacteria was measured and found to be 41%.

[0070] (Evaluation of dispersibility) Except for not mixing lactic acid bacteria, fats and oils and emulsifiers were mixed in the same manner as the lactic acid bacteria-containing self-emulsifying composition used in measuring survival rate, to prepare a composition for dispersibility evaluation (emulsifier ratio in composition 66.7 wt%, emulsifier content in composition 66.7 wt%, fats and oils content in composition 33.3 wt%).1 g of the prepared composition was suspended in 30 mL of water and allowed to stand, and when checking dispersibility in water, no separation occurred even after 3 hours of standing.

[0071] (Comparative Example 10) (Measurement of Viability of Lactic Acid Bacteria) A lactic acid bacteria-containing self-emulsifying composition was prepared in the same manner as in Example 1, using a glycerin fatty acid ester (Sunsoft Q-17D, manufactured by Taiyo Kagaku Co., Ltd.) as an emulsifier (initial number of bacteria: 4.5 × 10 １１ cfu / g, emulsifier ratio in the composition 66.7% by weight, emulsifier content in the composition 63.5% by weight, oil and fat content in the composition 31.7% by weight.

[0072] The obtained self-emulsifying composition containing lactic acid bacteria was stored at 25°C for 20 days in the same manner as in Example 1, and the survival rate of the lactic acid bacteria was measured and found to be 19%.

[0073] (Evaluation of Dispersibility) A composition for dispersibility evaluation was prepared by mixing fats and oils and an emulsifier in the same manner as the lactic acid bacteria-containing self-emulsifying composition used in measuring the survival rate, except that lactic acid bacteria were not mixed in (emulsifier ratio in the composition: 66.7 wt%, emulsifier content in the composition: 66.7 wt%, fats and oils content in the composition: 33.3 wt%). 1 g of the prepared composition was suspended in 30 mL of water and allowed to stand to check its dispersibility in water. Separation occurred within 1 to 3 hours after standing.

[0074] (Comparative Example 11) (Measurement of Viability of Lactic Acid Bacteria) A self-emulsifying composition containing lactic acid bacteria was prepared in the same manner as in Example 1, using polyglycerol condensed ricinoleic acid ester (SY Glystar CR-350H, manufactured by Sakamoto Pharmaceutical Co., Ltd.) as an emulsifier (initial cell count: 4.5 × 10 １１ cfu / g, emulsifier ratio in the composition 50.0% by weight, emulsifier content in the composition 47.6% by weight, oil and fat content in the composition 47.6% by weight.

[0075] The obtained self-emulsifying composition containing lactic acid bacteria was stored at 25°C for 20 days in the same manner as in Example 1, and the survival rate of the lactic acid bacteria was measured and found to be 94%.

[0076] (Evaluation of Dispersibility) A composition for dispersibility evaluation was prepared by mixing fats and oils and an emulsifier in the same manner as the lactic acid bacteria-containing self-emulsifying composition used in measuring the survival rate, except that lactic acid bacteria were not mixed in (emulsifier ratio in the composition: 50.0 wt%, emulsifier content in the composition: 50.0 wt%, fats and oils content in the composition: 50.0 wt%). 1 g of the prepared composition was suspended in 30 mL of water and allowed to stand to check its dispersibility in water. Separation occurred within 1 to 3 hours after standing.

[0077] Table 1 below summarizes the test results for the survival rate of lactic acid bacteria in Examples 1 and 2 and Comparative Examples 1 to 3.

[0078]

[0079] Tables 2 and 3 below summarize the test results for the survival rate and dispersibility in water of lactic acid bacteria in Examples 1 and 2 and Comparative Examples 4 to 11.

[0080]

[0081]

[0082] In Tables 2 and 3, "◯ (circle)" indicates that when the composition for dispersibility evaluation was suspended in water and allowed to stand, no separation occurred even after 3 hours of standing. "× (cross)" indicates that when the composition for dispersibility evaluation was suspended in water and allowed to stand, separation occurred within 1 to 3 hours of standing.

[0083] As shown in Tables 1 to 3, when a combination of DO-100V (monooleic acid diglyceride) and Tween 80 (polysorbate 80) or a combination of DO-100V and ML-750 (polyglycerol 10-laurate) was used as an emulsifier, a lactic acid bacteria survival rate of over 60% was achieved at an emulsifier ratio of 50% by weight, and the dispersibility in water was excellent (Examples 1 and 2). In contrast, when DO-100V alone was used as an emulsifier, the lactic acid bacteria survival rate and dispersibility in water decreased (Comparative Examples 1 and 5). When Tween 80 alone was used as an emulsifier, it was necessary to increase the emulsifier ratio to achieve a lactic acid bacteria survival rate similar to that of Examples 1 and 2 (Comparative Examples 2 and 6). When ML-750 alone was used as an emulsifier, the lactic acid bacteria survival rate was not as high as that of Examples 1 and 2, even when the emulsifier ratio was increased (Comparative Examples 3 and 7). When a combination of DO-100V and MM-750 was used as an emulsifier, the survival rate of lactic acid bacteria decreased (Comparative Example 4). When MM-750 alone was used as an emulsifier, the survival rate of lactic acid bacteria was only comparable to or lower than that of Examples 1 and 2, even when the emulsifier ratio was increased (Comparative Example 8). When MO-5S was used as an emulsifier, the survival rate of lactic acid bacteria was not as high as that of Examples 1 and 2, even when the emulsifier ratio was increased (Comparative Example 9). When Q-17D was used as an emulsifier, the survival rate of lactic acid bacteria was not as high as that of Examples 1 and 2, even when the emulsifier ratio was increased, and the dispersibility in water also decreased (Comparative Example 10). When CR-350H was used as an emulsifier, the dispersibility in water decreased (Comparative Example 11). Therefore, it was demonstrated that a probiotic-containing self-emulsifying composition containing a combination of (i) monooleic acid diglyceride and (ii) polysorbate or polyglycerol laurate as emulsifiers can achieve high probiotic survival rate while reducing the emulsifier ratio, and has excellent dispersibility in water.

[0084] (Examples 3 to 6) In Example 1, the ratio of DO-100V to Tween 80 in the composite emulsifier and the ratio of the composite emulsifier in the lactic acid bacteria-containing self-emulsifying composition were changed to prepare lactic acid bacteria-containing self-emulsifying compositions, and the effect on the survival rate of lactic acid bacteria was confirmed. The results are shown in Table 4.

[0085]

[0086] As is clear from Table 4, there was no significant change in the effect of the composite emulsifier at the DO-100V:Tween 80 ratio of 2:1 to 1:2 investigated and at composite emulsifier ratios of 33 to 50% in the lactic acid bacteria-containing self-emulsifying composition.

[0087] All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety.

Claims

1. A composition containing probiotics, an emulsifier, and an oil or fat, wherein the emulsifier is either 1) a complex emulsifier containing monooleic acid diglyceride and polysorbate, or 2) a complex emulsifier containing monooleic acid diglyceride and polyglycerol laurate.

2. The composition of claim 1, wherein the probiotic is a lactic acid bacterium.

3. The composition according to claim 1, wherein the weight ratio of monooleic acid diglyceride to polysorbate in the complex emulsifier is 2:1 to 1:

2.

4. The composition according to claim 1, wherein the weight ratio of monooleic acid diglyceride to polyglycerol laurate in the complex emulsifier is 2:1 to 1:

2.

5. The composition according to claim 1, wherein the weight ratio of said oil to said complex emulsifier is 7:3 to 2:

3.

6. The composition of claim 1, wherein the polysorbate is polysorbate 80.

7. The composition of claim 1, wherein said polyglycerol laurate is an ester of polyglycerol 10 and lauric acid.

8. The composition according to claim 1, wherein the oil is MCT (medium chain triglyceride).

9. The composition according to claim 1, which is used as an additive for food, drink or cosmetics.

10. The composition according to claim 1, which is a probiotic-containing food or drink or a probiotic-containing cosmetic.

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