Preparation method of soybean whey protein-hyaluronic acid stable emulsion

Abstract

The invention discloses a preparation method of soybean whey protein-hyaluronic acid stable emulsion, and belongs to the technical field of food processing. The method mainly comprises the following steps: 1) extracting soybean whey protein; 2) preparing a soybean whey protein-hyaluronic acid composite solution; 3) preparing the soybean whey protein-hyaluronic acid stable emulsion.The raw materials used in the preparation method are extracted from whey waste water generated by soybean product processing, and the preparation method has important significance in reducing resource waste and practicing the green health concept and meanwhile shows wide prospects in application in the fields of emulsion product development, drug delivery, food, cosmetics and the like.

Description

Technical Field

[0001] The invention relates to a preparation method of soybean whey protein-hyaluronic acid stable emulsion Background Art

[0002] Soy whey is a by-product of soybean processing and contains a variety of bioactive proteins, mainly soy whey protein, β-amylase, soybean trypsin inhibitor, lectin, etc. Natural soy whey protein has good solubility, foaming and dispersibility, but the emulsion prepared with soy whey protein alone often has some limitations, such as unsatisfactory emulsification stability, easy oil phase aggregation and stratification during storage, and difficulty in maintaining the stability of the emulsion system.

[0003] Hyaluronic acid is a natural linear polysaccharide with excellent gelling, thickening, hydrating and special physiological functions. Introducing hyaluronic acid into the emulsion system can adjust the interfacial properties of the emulsion, increase the strength of the interfacial film or adjust the overall viscosity of the system by interacting with the system components, thereby improving the stability of the emulsion.

[0004] Proteins and polysaccharides can form complexes through electrostatic interactions, hydrogen bonds, hydrophobic interactions, and other methods. During the preparation of the emulsion, these complexes are adsorbed and arranged at the oil-water interface to form an interfacial film with certain strength and elasticity. This interfacial film can effectively prevent the aggregation of oil droplets and the stratification of the emulsion, thereby improving the stability of the emulsion. At the same time, different combinations of proteins and polysaccharides, different proportions, and different preparation conditions will affect the formation of the complex and the performance of the emulsion, which also provides a theoretical basis for optimizing the performance of the emulsion by adjusting these parameters. It is of great significance to reduce resource waste and practice the concept of green health. At the same time, its application in the fields of emulsion product development, drug delivery, food and cosmetics shows broad prospects. Summary of the invention

[0005] The object of the present invention is to provide a method for preparing an emulsion using soy whey protein-hyaluronic acid to obtain a high-performance emulsion product suitable for use in multiple fields such as food and cosmetics.

[0006] To achieve the above objectives, the problem to be solved by the present invention is achieved through the following technical solutions:

[0007] 1. A method for preparing a soy whey protein-hyaluronic acid stable emulsion, characterized in that it comprises the following steps:

[0008] (1) Defatted soy flour and deionized water were mixed in a ratio of 1:10 (w / v), the pH was adjusted to 8.0, and the mixture was stirred for 2 h before centrifugation (8000 rpm, 30 min) to collect the supernatant; the pH of the supernatant was adjusted to 4.5, and the mixture was stirred for 30 min before centrifugation (8000 rpm, 20 min) to collect the supernatant, which was the crude soy whey protein solution; ammonium sulfate was added to the crude soy whey protein solution to reach 80% saturation, and then centrifuged (8000 rpm, 30 min); the centrifuged precipitate was redissolved in deionized water and dialyzed (MWCO 3500 Da, 4 ℃) for 48 h, with the water changed every 4-6 h; the dialyzate was freeze-dried for 24 h to obtain soy whey protein;

[0009] (2) Prepare a 20 mg / mL soy whey protein solution, stir for 2 h, hydrate in a 4 °C refrigerator overnight, and adjust the solution pH to 7.0 after returning to room temperature; prepare a 10 mg / mL hyaluronic acid solution, wait for it to completely dissolve, and then add it to the soy whey protein solution;

[0010] (3) Camellia oil was added to the solution containing soy whey protein and hyaluronic acid in proportion, and the mixture was stirred thoroughly at room temperature using a magnetic stirrer. After being homogenized at 10,000 r / min for 2 min, the mixture was ultrasonically treated to obtain a stable emulsion.

[0011] Preferably, the molecular weight of the hyaluronic acid is 100-200 kDa, 400-800 kDa, and 1500-2500 kDa, respectively.

[0012] Preferably, the mixing ratio of the hyaluronic acid solution to the soy whey protein solution is 1:1.

[0013] Preferably, the volume of the added camellia oil is 40%.

[0014] Preferably, the ultrasonic power of the ultrasonic treatment is 400 W and the ultrasonic time is 15 min.

[0015] The method for preparing emulsion by using soybean whey protein and hyaluronic acid of different molecular weights of the present invention has the advantages of:

[0016] 1) The soy whey protein in the present invention is extracted from a byproduct of soybean processing, which has a wide source and low cost. Its processing and utilization not only helps to increase the added value of the soybean protein industry, but also conforms to the current market trend of green consumption and sustainable development.

[0017] 2) The present invention compounds soybean whey protein with hyaluronic acid, which can maintain good stability of the emulsion and is more conducive to its practical application in the fields of food, cosmetics, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The average particle size of soy whey protein-hyaluronic acid composite emulsion;

[0019] Figure 2 The results are the emulsification activity and emulsification stability of soy whey protein-hyaluronic acid composite emulsion;

[0020] Figure 3 This is the creaming index result of soy whey protein-hyaluronic acid composite emulsion. DETAILED DESCRIPTION

[0021] The embodiments of the present invention are part of the embodiments of the present invention, and the protection scope of the present invention includes but is not limited to the following implementation examples. Example 1

[0022] 1. A method for preparing a soy whey protein-hyaluronic acid stable emulsion, characterized in that it comprises the following steps:

[0023] Defatted soy flour and deionized water were mixed in a ratio of 1:10 (w / v), the pH was adjusted to 8.0, and the mixture was stirred for 2 h before centrifugation (8000 rpm, 30 min) to collect the supernatant; the pH of the supernatant was adjusted to 4.5, and the mixture was stirred for 30 min before centrifugation (8000 rpm, 20 min) to collect the supernatant, which was the crude soy whey protein solution; ammonium sulfate was added to the crude soy whey protein solution to reach 80% saturation, and then centrifuged (8000 rpm, 30 min); the centrifuged precipitate was redissolved in deionized water and dialyzed (MWCO 3500 Da, 4 ℃) for 48 h, with the water changed every 4-6 h; the dialyzate was freeze-dried for 24 h to obtain soy whey protein; a soy whey protein solution with a concentration of 20 mg / mL was prepared, and after stirring for 2 h, the soy whey protein solution was concentrated at 4 ℃ refrigerator overnight, and after returning to room temperature, the solution pH was adjusted to 7.0; camellia oil was added to the solution containing soy whey protein at a volume ratio of 40%, and a magnetic stirrer was used to fully stir at room temperature; after rough averaging at 10000 r / min for 2 min, the final stable emulsion was treated at an ultrasonic power of 400 W for 15 min. When only soy whey protein was stable in the emulsion, the average particle size of the emulsion was large, 4.73±0.23μm, and the emulsion stability was poor. Example 2

[0024] 1. A method for preparing a soy whey protein-hyaluronic acid stable emulsion, characterized in that it comprises the following steps:

[0025] Defatted soy flour and deionized water were mixed in a ratio of 1:10 (w / v), the pH was adjusted to 8.0, and the mixture was stirred for 2 h and then centrifuged (8000 rpm, 30 min) to collect the supernatant; the pH of the supernatant was adjusted to 4.5, and the mixture was stirred for 30 min and then centrifuged (8000 rpm, 20 min) to collect the supernatant, which was the crude soy whey protein solution; ammonium sulfate was added to the crude soy whey protein solution to reach 80% saturation, and then centrifuged (8000 rpm, 30 min); the centrifuged precipitate was redissolved in deionized water and dialyzed (MWCO 3500 Da, 4 ℃) for 48 h, with the water changed every 4-6 h; the dialyzate was freeze-dried for 24 h to obtain soy whey protein; a soy whey protein solution with a concentration of 20 mg / mL was prepared, stirred for 2 h, hydrated in a refrigerator at 4 ℃ overnight, and the pH of the solution was adjusted to 7.0 after returning to room temperature; a soy whey protein solution with a concentration of 10 mg / mL and a molecular weight of 100-200 kDa hyaluronic acid solution was added to the soy whey protein solution at a ratio of 1:1 after it was completely dissolved; camellia oil was added to the solution containing soy whey protein and hyaluronic acid at a ratio of 40% by volume, and was fully stirred at room temperature using a magnetic stirrer; after coarse homogenization at 10000r / min for 2 min, the final stable emulsion was treated at an ultrasonic power of 400 W for 15 min. Example 3

[0026] 1. A method for preparing a soy whey protein-hyaluronic acid stable emulsion, characterized in that it comprises the following steps:

[0027] Defatted soy flour and deionized water were mixed in a ratio of 1:10 (w / v), the pH was adjusted to 8.0, and the mixture was stirred for 2 h and then centrifuged (8000 rpm, 30 min) to collect the supernatant; the pH of the supernatant was adjusted to 4.5, and the mixture was stirred for 30 min and then centrifuged (8000 rpm, 20 min) to collect the supernatant, which was the crude soy whey protein solution; ammonium sulfate was added to the crude soy whey protein solution to reach 80% saturation, and then centrifuged (8000 rpm, 30 min); the centrifuged precipitate was redissolved in deionized water and dialyzed (MWCO 3500 Da, 4 ℃) for 48 h, with the water changed every 4-6 h; the dialyzate was freeze-dried for 24 h to obtain soy whey protein; a soy whey protein solution with a concentration of 20 mg / mL was prepared, stirred for 2 h, hydrated in a refrigerator at 4 ℃ overnight, and the pH of the solution was adjusted to 7.0 after returning to room temperature; a soy whey protein solution with a concentration of 10 mg / mL and a molecular weight of 400-800 was prepared. kDa hyaluronic acid solution, after it is completely dissolved, is added to the soy whey protein solution at a ratio of 1:1; camellia oil is added to the solution containing soy whey protein and hyaluronic acid at a volume ratio of 40%, and is fully stirred at room temperature using a magnetic stirrer; after being roughly homogenized at 10000r / min for 2 min, the final stable emulsion is treated at an ultrasonic power of 400 W for 15 min. When hyaluronic acid with a molecular weight of 400-800 kDa is added to the system, the stability of the emulsion is improved, and the emulsifying activity is 24.98±0.14 m 2 / g, the emulsification stability was 43.85±0.41 min, and the creaming index was 11.15±0.89%, indicating that the emulsion stability was good. Example 4

[0028] 1. A method for preparing a soy whey protein-hyaluronic acid stable emulsion, characterized in that it comprises the following steps:

[0029] Defatted soy flour and deionized water were mixed in a ratio of 1:10 (w / v), the pH was adjusted to 8.0, and the mixture was stirred for 2 h and then centrifuged (8000 rpm, 30 min) to collect the supernatant; the pH of the supernatant was adjusted to 4.5, and the mixture was stirred for 30 min and then centrifuged (8000 rpm, 20 min) to collect the supernatant, which was the crude soy whey protein solution; ammonium sulfate was added to the crude soy whey protein solution to reach 80% saturation, and then centrifuged (8000 rpm, 30 min); the centrifuged precipitate was redissolved in deionized water and dialyzed (MWCO 3500 Da, 4 ℃) for 48 h, with the water changed every 4-6 h; the dialyzate was freeze-dried for 24 h to obtain soy whey protein; a soy whey protein solution with a concentration of 20 mg / mL was prepared, stirred for 2 h, hydrated in a refrigerator at 4 ℃ overnight, and the pH of the solution was adjusted to 7.0 after returning to room temperature; a soy whey protein solution with a concentration of 10 mg / mL and a molecular weight of 1500-2500 was prepared. kDa hyaluronic acid solution, after it is completely dissolved, is added to the soy whey protein solution at a ratio of 1:1; camellia oil is added to the solution containing soy whey protein and hyaluronic acid at a volume ratio of 40%, and is fully stirred at room temperature using a magnetic stirrer; after being roughly homogenized at 10000 r / min for 2 min, the final stable emulsion is treated at an ultrasonic power of 400 W for 15 min. When hyaluronic acid with a molecular weight of 1500-2500 kDa is added to the system, the stability of the emulsion is improved, the emulsification activity is 25.06±0.32m2 / g, the emulsification stability is 44.27±0.52 min, and the creaming index is reduced to 9.64±1.32%.

Claims

1. A method for preparing a soy whey protein-hyaluronic acid stable emulsion, characterized in that: The following steps are involved: (1) Defatted soy flour and deionized water were mixed in a ratio of 1:10 (w / v), the pH was adjusted to 8.0, and the mixture was stirred for 2 h before centrifugation (8000 rpm, 30 min) to collect the supernatant; the pH of the supernatant was adjusted to 4.5, and the mixture was stirred for 30 min before centrifugation (8000 rpm, 20 min) to collect the supernatant, which was the crude soy whey protein solution; ammonium sulfate was added to the crude soy whey protein solution to reach 80% saturation, and then centrifuged (8000 rpm, 30 min); the centrifuged precipitate was redissolved in deionized water and dialyzed (MWCO 3500 Da, 4 ℃) for 48 h, with the water changed every 4-6 h; the dialyzate was freeze-dried for 24 h to obtain soy whey protein; (2) Prepare a 20 mg / mL soy whey protein solution, stir for 2 h, hydrate in a 4 °C refrigerator overnight, and adjust the solution pH to 7.0 after returning to room temperature; prepare a 10 mg / mL hyaluronic acid solution, wait for it to completely dissolve, and then add it to the soy whey protein solution; (3) Camellia oil was added to the solution containing soy whey protein and hyaluronic acid in proportion, and the mixture was stirred thoroughly at room temperature using a magnetic stirrer. After being homogenized at 10,000 r / min for 2 min, the mixture was ultrasonically treated to obtain a stable emulsion.

2. A method for preparing a soy whey protein-hyaluronic acid stable emulsion according to claim 1, characterized in that: The molecular weights of the hyaluronic acid described in step (2) are 100-200 kDa, 400-800 kDa, and 1500-2500 kDa, respectively.

3. A method for preparing a soy whey protein-hyaluronic acid stable emulsion according to claim 1, characterized in that: In step (2), the mixing ratio of the hyaluronic acid solution to the soy whey protein solution is 1:

1.

4. A method for preparing a soy whey protein-hyaluronic acid stable emulsion according to claim 1, characterized in that: The volume of camellia oil added in step (3) is 40%.

5. A method for preparing a soy whey protein-hyaluronic acid stable emulsion according to claim 1, characterized in that: The ultrasonic power of the ultrasonic treatment described in step (3) is 400 W, and the ultrasonic time is 15 min.

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