KR101827611B1 - Water-in-oil micro or nanoemulsion comprising phycocyanin - Google Patents

Abstract

The present invention relates to a micro- or nano-emulsion including a phycocyanin, and more particularly to a micro-nano-emulsion comprising a phycocyanin, a surfactant, an auxiliary surfactant, an oil and water, and a cosmetic composition Or a functional food composition.

Description

[0002] WATER-IN-OIL MICRO OR NANOEMULSION COMPRISING PHYCOCYANIN < RTI ID = 0.0 >

The present invention relates to a water-in-oil micro- or nano-emulsion comprising phycocyanin, a process for producing the same, and a functional cosmetic composition and functional food composition comprising the emulsion.

Spirulina (Spirulina) is a kind of cyanobacteria, high in protein content, contains a large amount of minerals, vitamins, and fiber, and has become a future food resource for mankind. C-phycocyanin is a major picofile protein of Spirulina. It is a substance with antioxidant properties that is blue and melts in water to show fluorescence. Many studies have been conducted on picosanoids based on its antioxidant function, and various activities such as anti-inflammation, anticancer, hepatoprotective, and neuroprotective activity have been revealed (Non-Patent Documents 1 to 2) .

Pycocyanin effectively removes harmful free radicals in the body, preventing lipid peroxidation and inhibiting inflammation. It also has anti-inflammatory effects by reducing the activity of cyclooxygenase-2, an enzyme that produces inflammatory substances, and by inhibiting the release of histamine from mast cells. In addition, lipid peroxidation and inhibition of cyclooxygenase-2 can inhibit hepatotoxicity caused by carbon tetrachloride.

However, phycocyanin is sensitive and unstable to light, temperature, pH and moisture, and is thus not easy to use for skin or oral delivery by itself. Particularly, since picosian is a water-soluble component, it is difficult to permeate the stratum corneum which is the outermost part of the skin showing hydrophobicity and be absorbed into the skin. Therefore, there is a need for a method for effective percutaneous delivery of phycocyanin.

 C. Romay, R. Gonzalez, N. Ledon, D. Remirez, and V. Rimbau ... Curr.Protein.Pept.Sc., 4, 207-216 (2003)  B. Fernandez-Rojas, J. Hernandez-Juarez, and J. Pedraza-Chaverri ... J. Funct.Foods., 375-392 (2014)

SUMMARY OF THE INVENTION The present invention has been made in order to solve the problems of the prior art,

Which is capable of increasing the mobility, absorption and stability of the phycocyanin in the body, and a process for producing the microcrystalline microemulsion or nanoemulsion containing phycocyanin.

It is another object of the present invention to provide a functional cosmetic composition and a functional food composition containing the emulsion and being suitable for use in cosmetics or foods.

In order to solve the above problems,

A water-in-oil micro- or nanoemulsion comprising a surfactant, a phycocyanin, a surfactant, a co-surfactant, an oil and water.

In addition,

(1) dissolving phycocyanin in water to prepare an aqueous phase;

(2) mixing the surfactant and the cosurfactant;

(3) mixing oil into the mixture prepared in the step (2);

(4) mixing the mixture prepared in the step (3) and the aqueous phase of the step (1);

(5) stirring the mixture prepared in the step (4) to prepare a water-in-oil micro or nano emulsion.

The present invention also provides a functional cosmetic composition or a functional food composition comprising the water-in-oil micro or nano emulsion.

The water-in-oil micro- or nano-emulsion containing phycocyanin according to the present invention can increase the bioavailability of phycocyanin by increasing the absorption, transmission and stability of phycocyanin.

In addition, the use of the functional cosmetic composition or the functional food composition containing the micro or nano emulsion of the present invention can provide an excellent body absorption effect of picocyanin.

FIG. 1 is a flow chart showing a method for producing microcrystals or nanoemulsions containing phycocyanin by a magnetic coupling method as an example.
Figure 2 is a pseudo-ternary phase diagram of the nanoemulsion of Example 3 (A) and Example 4 (A) with different weight ratios of surfactant and co-surfactant.
3 is a photograph of a nano-emulsion of Example 1 observed with a transmission electron microscope.
4 is a graph showing the storage stability evaluation results of Examples 1 to 4.
5 (A) to 5 (D) are images obtained by observing skin slices after confocal laser scanning microscopy (CLSM) after curing for 4 hours in the skin absorption experiments of Examples 1 to 4, respectively.
6 (A) to (D) are images obtained by observing skin slices with a confocal laser scanning microscope (CLSM) after 8 hours of skin absorption experiments in Examples 1 to 4, respectively.

The present invention relates to a micro or nano emulsion comprising phycocyanin, a method for producing the same, and a functional cosmetic composition or functional food composition containing the emulsion.

The present inventors have found that when a water-in-oil type micro or nano emulsion including a phycocyanin, a surfactant, a cosurfactant, an oil and water is prepared and used, it has low stability and water solubility, And thus the present invention has been completed.

Hereinafter, the present invention will be described in detail.

As used herein, the term " active ingredient "refers to an ingredient that alone exhibits the desired activity or can exhibit activity together with a carrier that is itself inactive.

As used herein, the term "emulsion" refers to a micro- and / or nano-emulsion unless specifically limited.

The present invention provides a water-in-oil micro- or nano-emulsion comprising a phycocyanin, a surfactant, a co-surfactant, an oil and water.

Picosian

The phycocyanin contained in the emulsion of the present invention may be commercially available or may be produced by a manufacturing method including the following steps without limitation, and the form thereof may be a powder form or the like.

The above-

a) extracting spirulina with an extraction solvent;

b) centrifuging and filtering the extract obtained in step a); And

c) drying the filtrate obtained after the filtration.

In step (a), spirulina may be cultivated or commercially available. Examples of the extraction method include conventional extraction methods such as ultrasonic extraction, hot water extraction or reflux cooling extraction. The ultrasonic extraction method may be preferably repeated three to five times, but is not limited thereto.

The drying in the step c) may be freeze-drying using a freeze dryer, but is not limited thereto. In addition to freeze-drying, vacuum drying or vacuum drying is also possible.

The amount of phycocyanin is preferably 0.05 to 0.5% by weight, more preferably 0.1 to 0.2% by weight based on the total weight of the emulsion of the present invention.

Surfactants

The surfactant contained in the emulsion of the present invention minimizes the tension of the water phase and the oil phase and mixes the two phases to form micro or nano sized particles uniformly. As the surfactant, at least one surfactant selected from nonionic, anionic, cationic and amphoteric surfactants may be used, but nonionic surfactants may be more preferably used.

In the present invention, the total hydrophilic-lipophile balance (HLB) value of the surfactant may preferably be 4 to 11. The total hydrophilic lipophilic balance value can be calculated by a conventional method.

Specific examples of the surfactant include sorbitan monooleate (hereinafter also referred to as 'Span 80'), polyethylene glycol sorbitan monooleate (hereinafter also referred to as 'Tween 80'), sorbitan monostearate (Also referred to as 'Span 60'), and polyethylene glycol sorbitan monostearate (hereinafter also referred to as 'Tween 60'), and at least one selected therefrom can be used. More preferably, at least one selected from sorbitan monooleate (Span 80) and polyethylene glycol sorbitan monooleate (Tween 80) is used.

In a preferred embodiment of the present invention, when sorbitan monooleate (Span 80) and polyethylene glycol sorbitan monooleate (Tween 80) are used as the surfactant, the weight ratio is preferably 3: 7 to 6: 4 , And more preferably from 4: 6 to 5: 5. When the emulsion is used in such a weight ratio as above, the produced emulsion exhibits excellent transparency and phase separation does not occur.

The surfactant is preferably contained in an amount of 40 to 80% by weight, and more preferably 50 to 70% by weight, based on the total weight of the emulsion of the present invention.

Ancillary surfactant

Auxiliary surfactants included in the emulsions of the present invention help to produce more stable emulsions with surfactants and serve to impart additional properties such as moisturizing. The cosurfactant is preferably used because it exhibits a HLB value different from that of the above-mentioned surfactant, forming a more stable emulsion. In the present invention, the HLB value of the cosurfactant may be preferably 5 to 14.

Specific examples of the cosurfactant include propylene glycol, butylene glycol, and polyethylene glycol, and at least one selected from the above surfactants can be used. More preferably, propylene glycol can be used.

The weight ratio of the surfactant to the cosurfactant contained in the emulsion of the present invention is preferably 1: 1 to 7: 1, more preferably 2: 1 to 6: 1, more preferably 3: 1 to 5: 1 More preferable. When the weight ratio of the surfactant to the cosurfactant is within the above range, it is advantageous for the production of transparent micro or nano emulsion.

As an embodiment of the present invention, the pseudo-tertiary phase diagrams of Examples 3 to 4 are shown in FIG. 2, when the weight ratio of the surfactant to the cosurfactant is 3: 1 (A in FIG. 2), 5: 1 (Example B in FIG. 2) The transparent micro or nanoemulsion formation area was the largest.

 The auxiliary surfactant is preferably contained in an amount of 10 to 20% by weight, and more preferably 11 to 18% by weight, based on the total weight of the emulsion of the present invention.

oil

The oil to be contained in the emulsion of the present invention is not limited to a great extent, but it is preferable to use at least one selected from vegetable oil which can be added to food and is safe oil for skin. However, since the oil phase may separate during the preparation of the emulsion with the longer chain fat, it may be more preferable to use a heavy chain fat. The heavy chain fat is absorbed directly without being decomposed upon ingestion, and has the effect of reducing the anti-cancer effect and the risk of heart disease, and it can be rapidly absorbed to the skin to enhance the effect of supplying nutrients and preventing aging.

Specific examples of the oil include caprylic / capric triglyceride, olive oil, castor oil, safflower oil, soybean oil, and vegetable oil rapeseed oil and the like, and at least one selected from these may be used. More preferably, caprylic / capric triglyceride may be used. The caprylic / capric triglyceride is a triester mixture of caprylic acid and capric acid and glycerin, which is commercially available under the trade name Miglyol 812 (Cremer Oleo GmbH & Co. KG, Germany) It can also be used.

The oil is preferably contained in an amount of usually 10 to 30% by weight, and more preferably 20 to 30% by weight, based on the total weight of the emulsion of the present invention.

water

The water contained in the emulsion of the present invention is not particularly limited, and water of any source can be used. It is also possible to use different types of water, preferably purified water such as demineralized water, brine or water with added additives. More preferably, purified water can be used.

Water is preferably contained in an amount of usually 5 to 20% by weight, and more preferably 8 to 15% by weight, based on the total weight of the emulsion of the present invention.

The emulsion of the present invention may further contain other ingredients such as a stabilizer, a moisturizer, a flavor (flavoring agent), a preservative or an antioxidant, if necessary within the scope of the present invention, in addition to the above-mentioned ingredients.

The present invention also provides a method for producing a water-in-oil micro- or nano-emulsion containing phycocyanin using the above-described components.

In the above production method,

(1) dissolving phycocyanin in water to prepare an aqueous phase;

(2) mixing the surfactant and the cosurfactant;

(3) mixing oil into the mixture prepared in the step (2);

(4) mixing the mixture prepared in the step (3) and the aqueous phase of the step (1);

(5) stirring the mixture prepared in the step (4) to prepare a water-in-oil micro or nano emulsion.

In the step (1), the phycocyanin is preferably used in an amount of 0.5 to 5 parts by weight, more preferably 1 to 2 parts by weight, based on 100 parts by weight of water. The formulation of phycocyanin to be used at this time is not particularly limited, but a powder form can be used from the viewpoint of ease of use.

In the mixing of the steps (2) and (3), it is preferable that the mixture is sufficiently stirred so that the interface between the mixture does not appear.

In the mixing of the step (4), it is preferable that the aqueous phase is slowly added dropwise to the mixture prepared in the step (3) in that stable micro or nano emulsion can be produced. In addition, micro- or nano-emulsions can be produced by a self-assembly method through this method.

The stirring speed in the step (5) is preferably 1,000 to 1,400 rpm, more preferably 1,200 to 1,300 rpm. When the stirring speed is within the above-mentioned range, the mixing is uniform and the oil-in-water type emulsion can be easily produced. The stirring is preferably performed for 3 to 5 minutes, more preferably 4 minutes, but is not limited thereto.

The emulsion preparation process of the present invention including the steps (1) to (5) may be carried out usually at room temperature.

The present invention also provides a functional cosmetic composition or a functional food composition comprising the micro or nano emulsion of the present invention.

The cosmetic composition may be manufactured by, but not limited to, a gel, a lotion, a powder, a lotion, a shampoo, a rinse, a cleanser, a cream or an essence. If necessary, an emulsifier, preservative, pH adjuster, perfume or the like may be further added to prepare a cosmetic preparation.

The functional food composition may be prepared by powders, granules, tablets, capsules, syrups, beverages, chewing gums, candies, tea, ice creams or confectionery, but is not limited thereto. In addition, if necessary, other ingredients may be further added in a necessary amount to prepare the food according to the conventional method for producing a functional food.

The functional cosmetic composition or the functional food prepared using the functional cosmetic composition or the functional food composition can stably preserve the picocyanin component for a long period of time, increase the absorption rate in the body, and provide the effect of enhancing the picocyanin in the body.

Hereinafter, the present invention will be described in more detail with reference to Examples and Comparative Examples. However, the following examples illustrate the present invention and the present invention is not limited by the following examples, and various modifications and changes may be made. The scope of the present invention will be determined by the technical idea of the following claims.

< Example >

Example  1-4. Picocyanin-containing Nano emulsion  Produce

The nanoemulsions of Examples 1 to 4 were prepared with the composition shown in Table 1 below.

The water phase was prepared by dissolving phycocyanin powder (phycocyanin extracted from Spirulina maxima, supplied by JC Creation) in purified water at a concentration of 10 mg / g.

Surfactants Span 80 and Tween 80 and the cosurfactant Propylene glycol were thoroughly stirred at 25 ° C for 60 minutes and mixed with oil caprylic / capric triglyceride (Miglyol® 812) and stirred again. The water phase thus prepared was slowly added dropwise and stirred at 1,200 rpm for 4 minutes to prepare a water-in-oil type nanoemulsion containing phycocyanin (FIG. 1). At this time, the nano emulsion was made by a self-assembly method.

Item Mixing ratio Content (% by weight) S80: T80 (S80 + T80): PG Picosian Purified water Span
80 Tween
80 PG Miglyol®
812 Example 1 1: 1 3: 1 0.1 10 26.2 26.2 17.5 20 Example 2 1: 1 5: 1 0.1 10 29.1 29.1 11.7 20 Example 3 1: 1.5 3: 1 0.1 10 21 31.5 17.5 20 Example 4 1: 1 3: 1 0.1 10 22.5 22.5 15 30

week)

S80: Span 80 (manufactured by Samchun Pure Chemical)

T80: Tween 80 (manufactured by Samchun Pure Chemical)

PG: Propylene glycol

Miglyol 812 (manufacturer: Cremer Oleo GmbH & Co. KG, Germany)

< Test Example >

Test Example  1. Microcrystals containing phycocyanin or Nano-emulsion  Physical property measurement experiment

The particle size, polydispersity index (PDI) and viscosity of the microspheres or nanoemulsions containing phycocyanin prepared in Examples 1 to 4 were measured to confirm their physical properties.

The particle size and polydispersity index of the micro or nano emulsion were measured using a dynamic light scattering apparatus (Malvern Zetasizer Nano ZE, Malvern Instruments, Malvern, U.K.). The viscosity was also measured using a viscometer (DV-II viscometer, Brookfield, USA). The results are shown in Table 2 below.

In all of Examples 1 to 4, the particle size was measured to be 100 nm or less, and Example 3 was the smallest at about 49 nm. Viscosity was the lowest at 205 CPS in Example 4 where the surfactant was relatively low.

Test Example  2. Microcrystals containing phycocyanin or Nano-emulsion  Morphological observation

The morphology of the microspheres or nanoemulsions containing the phycocyanin prepared in Examples 1 to 4 was observed using a transmission electron microscope (TEM). The results of Example 1 are shown in Fig.

It was confirmed that the particles of the microspheres or the nano emulsions prepared in Examples 1 to 4 according to the present invention had a uniform spherical shape and formed a desirable water-in-oil type emulsion. The particle size showed a particle size of 100 nm or less similar to the size measured using the dynamic light scattering apparatus in Test Example 1 above.

Test Example  3. Microcrystals containing phycocyanin or Nano-emulsion  Stability test

To confirm the physical stability of the microspheres containing the phycocyanin prepared in Examples 1 to 4, centrifugation was carried out at 25 DEG C and 400 rpm for 30 minutes using a centrifuge (Beckman Abanti JE, Beckman) The results are shown in Table 2 below.

Test Example  4. Microcrystals containing phycocyanin or Nano-emulsion  Storage stability test

In order to evaluate the storage stability of microcapsules or nanoemulsions containing phycocyanin prepared in Examples 1 to 4, the particle size was stored in a dynamic light scattering apparatus (Malvern Zetasizer Nano ZE, Malvern Instruments, Malvern, UK), and the results are shown graphically in FIG.

4, it was confirmed that the emulsions of Examples 1 to 4 according to the present invention maintained a stable state without changing the particle size for 4 weeks.

Test Example  5. Microcrystals containing phycocyanin or Nano-emulsion In vitro (in-vitro) skin absorption measurement

Samples were prepared in the same manner as in the emulsion preparation methods of Examples 1 to 4 for in vitro skin absorption measurement of the microspheres or nanoemulsions containing phycocyanin prepared in Examples 1 to 4 , And a small amount of rhodamine B (rhodamine B), which is a fluorescent marker, was used instead of phycocyanin.

For the skin absorption test, the dorsal skin of a mini-pig, 6 to 8 months old, was sampled and mounted on a Franz-type diffusion cell (Dihan Labtech, Korea). A phosphate buffer solution (pH 7.4) was added to the container (15.9 ml) of the prase-type diffusion cell. The area of the skin that absorbed and diffused was 2.27 cm ² , and the diffusion cell was mixed and dispersed at 600 rpm while being maintained at 37 ± 1 ° C.

1 g of the nano emulsion containing the fluorescent labeling substance was put into each donor container. After the absorption of the micro or nano emulsion was completed, the skin was washed with dried Kimwipes or PBS (Phosphate buffer saline) solution. The washed skin was frozen at -26 ° C or lower using a solvent (O.C. T. compound), cut to a thickness of 20 μm and placed on a slide glass. The cover glass was immediately covered and observed with a confocal scanning laser microscope (CLSM).

In order to confirm the degree of skin absorption, a confocal laser scanning microscope (CLSM: LSM 700, Carl-Zeiss, Germany) was used and the results are shown in FIG. 5 and FIG.

As can be seen from the results of FIGS. 5 to 6, it can be seen that the micro or nano emulsion prepared in Examples 1 to 4 according to the present invention has excellent skin absorbency.

FIG. 5 is a photograph taken after 4 hours from the start of the skin permeation experiment, and it was observed that the micro or nano emulsion penetrates the stratum corneum and enters the skin layer.

FIG. 6 shows photographs taken after 8 hours from the start of the skin permeation test. It can be seen that the micro or nano emulsion is much superior to that of FIG. 5 in the degree of permeation. In Example 1 (FIG. 6A) The degree of transmission of Example 2 (FIG. 6B) was superior to that of Example 3 (FIG. 6C) and Example 4 (FIG. 6D).

Test Example  6. Microcrystals containing phycocyanin or Nano-emulsion  Transparency measurement

The results of microscopic observation of the transparency of the microcapsules or nanoemulsions containing the phycocyanin prepared in Examples 1 to 4 are shown in Table 2 below.

Item Particle Size (nm) The polydispersity index (PDI) Viscosity (CPS) Whether phase separation transparency Example 1 59.27 ± 3.18 0.296 288.50 ± 6.45 Does not occur Transparency Example 2 78.21 + - 3.48 0.297 365.67 ± 14.72 Does not occur Transparency Example 3 49.08 ± 1.77 0.397 240.33 + - 18.21 Does not occur Transparency Example 4 86.20 ± 2.32 0.272 205 ± 7.91 Does not occur Transparency

As can be seen from the results shown in Table 2, Examples 1 to 4, which are the nanofiber emulsion of the present invention prepared with a nanoscale particle size of 100 nm or less, exhibit excellent transparency and have a polydispersity index of 0.4 or less All were at a desirable level.

In addition, it was confirmed that phase separation did not occur even after centrifugation for 30 minutes, and thus stability was obtained. In the storage stability test with aging, there was little change in the particle size for 4 weeks, indicating that the aging stability was also excellent (Fig. 4).

As a result of measuring the skin absorbency of the water-in-oil emulsion containing phycocyanin of Examples 1 to 4, it was confirmed that the skin absorbency was excellent. Further, since the skin absorption of the fluorescent marker increases with time, it can be judged that the absorption of the phycocyanin, which exhibits water solubility, also increases with the lapse of time (FIGS. 5 and 6). Thus, it can be confirmed that the absorption of phycocyanin is achieved to a satisfactory level by using the micro or nano emulsion of the present invention.

Claims (15)

A surfactant, a cosurfactant, an oil and water, does not contain sodium alginate,
Wherein the surfactant comprises sorbitan monooleate and polyethylene glycol sorbitan monooleate in a weight ratio of 3: 7 to 6: 4,
Wherein the cosurfactant comprises propylene glycol,
Wherein the oil comprises caprylic / capric triglyceride,
The weight ratio of the surfactant to the co-surfactant is 3: 1 to 5: 1,
The surfactant and the co-surfactant have different Hydrophile-Lipophile Balance (HLB) values,
The total hydrophilic lipophilic balance value of the surfactant is 4 to 11,
0.05 to 0.5% by weight, based on the total weight of the emulsion, of picosan; 50 to 70% by weight of a surfactant; 10 to 20% by weight of cosurfactant; 10 to 30% by weight of an oil, and 5 to 20% by weight of water. delete delete delete delete delete delete delete delete delete (1) dissolving phycocyanin in water to prepare an aqueous phase;
(2) mixing the surfactant and the cosurfactant;
(3) mixing oil into the mixture of step (2);
(4) mixing the mixture prepared in the step (3) and the aqueous phase of the step (1);
(5) stirring the mixture prepared in the step (4)
Wherein the surfactant comprises sorbitan monooleate and polyethylene glycol sorbitan monooleate in a weight ratio of 3: 7 to 6: 4,
Wherein the cosurfactant comprises propylene glycol,
Wherein the oil comprises caprylic / capric triglyceride,
The weight ratio of the surfactant to the co-surfactant is 3: 1 to 5: 1,
The surfactant and the co-surfactant have different Hydrophile-Lipophile Balance (HLB) values,
The total hydrophilic lipophilic balance value of the surfactant is 4 to 11,
0.05 to 0.5% by weight, based on the total weight of the emulsion, of picosan; 50 to 70% by weight of a surfactant; 10 to 20% by weight of cosurfactant; 10 to 30% by weight of an oil, and 5 to 20% by weight of water,
Wherein the water-in-oil emulsion contains no sodium alginate. delete The method of claim 11,
Wherein the stirring speed of the step (5) is 1,000 to 1,400 rpm. A functional cosmetic composition comprising the emulsion of claim 1. A functional food composition comprising the emulsion of claim 1.

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