CN102085173B - Bioactive emulsion using natural fibroin as emulsifier and preparation method thereof - Google Patents

Abstract

A bioactive emulsion using natural silk protein as emulsifier is prepared through dissolving degummed silk in LiBr aqueous solution to obtain silk protein aqueous solution, dialysis, centrifugal treatment to remove aggregate, mixing with oil phase, and homogenizing. According to the invention, natural fibroin is used as an emulsifier to prepare the bioactive emulsion with strong interface stability and excellent biocompatibility, and the droplet particle size, stability and fluidity of the emulsion can be regulated and controlled by emulsion components and proportions; the method has the advantages of simple preparation method, low cost, good repeatability, strong controllability and the like, and has wide application prospect in the fields of cosmetics, biopharmaceuticals and the like.

Description

Bioactive emulsion using natural silk protein as emulsifier and preparation method thereof

technical field

The invention relates to the preparation of an emulsion in the technical field of biomaterials, in particular to a biologically active emulsion using natural silk protein as an emulsifier and a preparation method thereof.

Background technique

Emulsion is a kind of dispersion system composed of two immiscible liquids, which is widely used in food, cosmetics, medicine, chemical industry and other fields. In an emulsion, the phase that exists in the form of small droplets is called the dispersed phase or the internal phase, and the remaining phase is called the continuous phase or the external phase. The types of emulsions formed include oil-in-water (O/W) type and water-in-oil (W/ O) type. The uniform dispersion of one liquid into another immiscible liquid will generate many interfaces and interfacial tensions, and emulsifiers need to be added to stabilize the emulsion and form the required droplet size. Emulsifiers mainly perform two functions, one is to reduce the interfacial tension by reducing the free energy of emulsion formation, and the other is to form a protective film or barrier at the liquid/liquid interface to prevent or slow down droplet flocculation and coalescence. The types of emulsifiers include common ionic substances, colloidal solid materials, amphiphilic polymers and surfactants. With the continuous improvement of living standards, people's demand for green, environmental protection, safety and high-quality high-end products is also increasing. Traditional emulsifiers can no longer meet the needs of high-tech industry development. They have excellent biocompatibility and biological Natural emulsifiers with degradability and strong interfacial stability will become an urgent need and an inevitable trend in the development of emulsion technology, especially in the cosmetics and biopharmaceutical industries.

After searching the prior art, it was found that Chinese Patent Document No. CN1295440A, published on May 16, 2001, described a "functional protein composition, an emulsion based on the composition and its preparation method", which is to The lupine protein composition is mixed with oil and water to prepare lupine-based emulsions for the food industry. This method requires reconstitution pretreatment and purification posttreatment, and the process is complicated. Chinese Patent Document No. CN1392773A, published on January 22, 2003, describes a "whey protein emulsion". The method is to prepare an emulsion from whey protein and cream for use as a dairy additive.

However, most of the natural protein emulsions used in this prior art are used in the food industry, and the bioactive emulsions that can be applied to industries such as cosmetics and biopharmaceuticals are rarely involved, especially the biocompatibility that has been applied in the biomedical field. , Natural silk related products with excellent mechanical properties.

Contents of the invention

The present invention aims at the above-mentioned deficiencies existing in the prior art, and provides a biologically active emulsion using natural silk protein as an emulsifier and a preparation method thereof, using natural silk protein as an emulsifier to prepare a product with strong interfacial stability and excellent biocompatibility Bioactive emulsion, the droplet size, stability and fluidity of the emulsion can be regulated by the emulsion components and the ratio. The invention has the advantages of simple preparation method, low cost, good repeatability, strong controllability, etc., and has broad application prospects in the fields of cosmetics, biopharmaceuticals and the like.

The present invention is achieved through the following technical solutions:

The invention relates to a preparation method of a biologically active emulsion using natural silk protein as an emulsifier. The aqueous solution of silk protein is obtained by dissolving degummed silk in LiBr aqueous solution. The aggregates are removed by dialysis and centrifugation, mixed with oil phase, homogenized Emulsification treatment to obtain bioactive emulsion.

The degummed silk refers to natural silkworm cocoons soaked in sodium carbonate aqueous solution and boiled.

The ratio of the natural cocoons to the sodium carbonate aqueous solution is 1:50-150 by weight.

The weight percentage concentration of described sodium carbonate aqueous solution is 0.2-0.8wt%;

The boiling time is 0.5-1.5 hours.

The concentration of the LiBr aqueous solution is 9.3mol/L.

The melting temperature is 40-80°C;

Described dialysis adopts the deionized water of 50-150 times of LiBr aqueous solution volume;

The time of the dialysis is 24-72 hours.

The speed of the centrifugation is 3000-6000rpm, and the centrifugation time is 5-15 minutes;

Said mixing with the oil phase refers to: dilute the fibroin aqueous solution after dialysis and centrifugation with deionized water to a weight percent concentration of 0.25-2wt%, and then mix it with the oil phase at a volume ratio of 50:50-90:10.

The oil phase is one or a combination of fatty oil, silicone oil, esters or alcohols, preferably edible oil, mineral oil, polydimethylsiloxane, ethyl acetate or polyethylene glycol one or a combination thereof;

The homogeneous emulsification treatment refers to emulsifying for 2-5 minutes with a homogenizer at a speed of 8000-14000 rpm.

The invention relates to the bioactive emulsion prepared by the above method, which is an oil-in-water type, has high stability, and the average droplet diameter is about 10-70 μm.

The present invention has substantive features and significant progress, and the present invention has the following advantages and positive effects: 1. The preparation method is simple, efficient and repeatable. 2. The emulsion has high stability and is suitable for different oil phases. 3. The particle size, stability and fluidity of emulsion droplets are controllable. 4. The emulsion has good biocompatibility and can be widely used in cosmetics and biopharmaceutical industries. 5. Wide source of raw materials and low cost.

Detailed ways

The embodiments of the present invention are described in detail below. This embodiment is implemented on the premise of the technical solution of the present invention, and detailed implementation methods and specific operating procedures are provided, but the protection scope of the present invention is not limited to the following implementation example.

Example 1

The natural silkworm cocoons are put into 50 times the weight of 0.8wt% sodium carbonate aqueous solution and boiled for 0.5 hour for degumming treatment, then deionized washing and drying. The degummed silk was dissolved in 9.3M LiBr aqueous solution at 80° C., and then dialyzed with 50 times the volume of deionized water for 72 hours to obtain a silk protein aqueous solution. Centrifuge the silk protein aqueous solution at 6000rpm for 5 minutes to remove aggregates, then dilute to a concentration of 0.25wt% by weight, add olive oil at a volume ratio of 50:50 to oil and water, and use a homogenizer emulsification method to emulsify at 14000rpm for 2 The bioactive emulsion with silk protein as emulsifier is prepared in minutes.

The prepared bioactive emulsion is an oil-in-water type with high stability, and the average droplet diameter is about 30-70 μm.

Example 2

The natural silkworm cocoon is put into 0.5wt% sodium carbonate aqueous solution whose weight is 100 times and boiled for 1 hour for degumming treatment, then deionized washing and drying. The degummed silk was dissolved in 9.3M LiBr aqueous solution at 60° C., and then dialyzed with 100 times the volume of deionized water for 48 hours to obtain a silk protein aqueous solution. Centrifuge the silk protein aqueous solution at 5000rpm for 10 minutes to remove aggregates, then dilute to a weight percentage concentration of 1wt%, add polydimethylsiloxane with an oil-water volume ratio of 20:80, and use a homogenizer to emulsify at 10000rpm The bioactive emulsion with silk protein as emulsifier was prepared by emulsifying for 3 minutes under the condition.

The prepared bioactive emulsion is an oil-in-water type with high stability, and the average droplet diameter is about 20-40 μm.

Example 3

The natural cocoon is put into 0.2wt% sodium carbonate aqueous solution with a weight of 150 times and boiled for 1.5 hours for degumming treatment, then deionized washing and drying. The degummed silk was dissolved in 9.3M LiBr aqueous solution at 40°C, and then dialyzed with 150 times the volume of deionized water for 24 hours to obtain a silk protein aqueous solution. The silk protein aqueous solution was centrifuged at 3000rpm for 15 minutes to remove aggregates, then diluted to a concentration of 2wt% by weight, ethyl acetate was added with an oil-water volume ratio of 10:90, and emulsified at 8000rpm using a homogenizer for 5 The bioactive emulsion with silk protein as emulsifier is prepared in minutes.

The prepared bioactive emulsion is an oil-in-water type with high stability, and the average droplet diameter is about 10-30 μm.

Example 4

The natural silkworm cocoon is put into 0.5wt% sodium carbonate aqueous solution whose weight is 100 times and boiled for 1 hour for degumming treatment, then deionized washing and drying. The degummed silk was dissolved in 9.3M LiBr aqueous solution at 60° C., and then dialyzed with 100 times the volume of deionized water for 48 hours to obtain a silk protein aqueous solution. Centrifuge the silk protein aqueous solution at 5000rpm for 10 minutes to remove aggregates, then dilute to a weight percentage concentration of 1wt%, add polyethylene glycol at an oil-water volume ratio of 20:80, and emulsify at 10000rpm using a homogenizer emulsification method The bioactive emulsion with silk protein as emulsifier is prepared in 3 minutes.

The prepared bioactive emulsion is an oil-in-water type with high stability, and the average droplet diameter is about 10-30 μm.

Claims (10)

1. one kind is the method for preparing of the biological activity emulsion of emulsifying agent with natural silk albumen; It is characterized in that; Through being dissolved in, boiled silk obtains the fibroin aqueous solution in the LiBr aqueous solution; Mixes with oil phase after removing aggregate through the dialysis centrifugal treating, obtaining the biological activity emulsion through the emulsifying processing.

2. according to claim 1 is the method for preparing of the biological activity emulsion of emulsifying agent with natural silk albumen, it is characterized in that described boiled silk is meant: the natural Bombyx bombycis that soaks and boil through aqueous sodium carbonate.

3. according to claim 2 is the method for preparing of the biological activity emulsion of emulsifying agent with natural silk albumen, it is characterized in that the ratio of described natural Bombyx bombycis and aqueous sodium carbonate is weight ratio 1: 50-150.

According to claim 2 or 3 described be the method for preparing of the biological activity emulsion of emulsifying agent with natural silk albumen, it is characterized in that the weight percent concentration of described aqueous sodium carbonate is 0.2-0.8wt%.

5. according to claim 1 is the method for preparing of the biological activity emulsion of emulsifying agent with natural silk albumen, it is characterized in that the concentration of described LiBr aqueous solution is 9.3mol/L.

6. according to claim 1 is the method for preparing of the biological activity emulsion of emulsifying agent with natural silk albumen, it is characterized in that, the 50-150 deionized water doubly of LiBr aqueous solution volume is adopted in described dialysis.

7. according to claim 1 is the method for preparing of the biological activity emulsion of emulsifying agent with natural silk albumen; It is characterized in that described the mixing with oil phase is meant: the fibroin aqueous solution after the centrifugal treating of will dialysing adds deionized water, and to be diluted to weight percent concentration be to mix by 50: 50～90: 10 volume ratio with oil phase behind the 0.25-2wt%.

According to claim 1 or 7 described be the method for preparing of the biological activity emulsion of emulsifying agent with natural silk albumen, it is characterized in that described oil phase is a kind of of fatty oil, silicone oil, esters or pure apoplexy due to endogenous wind or its combination.

9. according to claim 1 is the method for preparing of the biological activity emulsion of emulsifying agent with natural silk albumen, it is characterized in that, described emulsifying is handled and is meant: adopting homogenizer is 8000-14000rpm emulsifying 2-5 minute with speed.

10. one kind is the biological activity emulsion of emulsifying agent with natural silk albumen, it is characterized in that, prepares through the said method of above any claim.