CN113208097A - Fish skin gelatin emulsion stabilized by sodium alginate and corn starch and preparation method thereof - Google Patents
Fish skin gelatin emulsion stabilized by sodium alginate and corn starch and preparation method thereof Download PDFInfo
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- CN113208097A CN113208097A CN202110520636.9A CN202110520636A CN113208097A CN 113208097 A CN113208097 A CN 113208097A CN 202110520636 A CN202110520636 A CN 202110520636A CN 113208097 A CN113208097 A CN 113208097A
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- fish skin
- sodium alginate
- corn starch
- skin gelatin
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/10—Foods or foodstuffs containing additives; Preparation or treatment thereof containing emulsifiers
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
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- A23L2/62—Clouding agents; Agents to improve the cloud-stability
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
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- A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
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Abstract
A fish skin gelatin emulsion stabilized by sodium alginate and corn starch and a preparation method thereof belong to the technical field of emulsion preparation. Firstly, mixing sodium alginate and corn starch, dissolving in deionized water, carrying out water bath reaction, and cooling to room temperature to obtain a biopolymer stabilizer; dissolving fishskin gelatin in a stabilizer to prepare a water phase, and adding fat-soluble active substances into the corn oil to serve as an oil phase; and mixing the water phase and the oil phase, and then dispersing at a high speed and homogenizing at a high pressure to obtain the fish skin gelatin emulsion with stable sodium alginate and corn starch. Compared with the emulsion prepared by only using fish skin gelatin under the same condition, the emulsion prepared by the invention is an oil-in-water delivery system which has the advantages of low cost, safety, convenience, higher BITC embedding rate and bioavailability and better storage stability, and has great potential in the development of health-care foods, functional beverages and other products.
Description
Technical Field
The invention belongs to the technical field of emulsion preparation, and particularly relates to a fish skin gelatin emulsion stabilized by sodium alginate and corn starch and a preparation method thereof.
Background
Fat-soluble active substances such as n-3 polyunsaturated fatty acid, fat-soluble vitamins, phytosterol and the like have high utilization value, but most of the fat-soluble active substances have the defects of poor water solubility, easy oxidative degradation, low bioavailability and the like, so that the application of the fat-soluble active substances in the field of food is greatly limited. How to introduce fat-soluble active substances into food systems becomes a key link in the development of novel functional foods. The O/W type emulsion system is adopted for embedding, so that the fat-soluble active substances can be effectively protected, and the bioavailability of the fat-soluble active substances can be improved by changing the composition and the structure of the emulsion.
An O/W emulsion is a heterogeneous liquid dispersion in which a liquid is dispersed in the form of droplets (dispersed phase, i.e., oil phase) in another liquid immiscible therewith (continuous phase, i.e., water phase). At the moment, the interfacial area is increased sharply, the free energy of the system is increased, a thermodynamically unstable system is formed, and a surfactant is added to reduce the free energy of the system. The protein is a macromolecular surfactant, contains hydrophilic groups, hydrophilic groups and hydrophilic groups, and can be adsorbed on an interface and directionally arranged to form an interface adsorption layer, so that the energy of the system is reduced, and the stability of a dispersion system is improved. The fish skin gelatin is high-quality marine protein with relatively high molecular weight, is obtained by hydrolyzing collagen of aquatic fishes, is a byproduct in the fishery processing process, is rich and economic in raw material acquisition, and has extremely high utilization value. Meanwhile, the fish skin gelatin has high surface activity, can form droplets with positive charges, and is widely used as an emulsifier in the food industry. Polysaccharides are also commonly used emulsifiers, and emulsions thereof are stable under a variety of environmental conditions. The combination of protein and polysaccharide may improve emulsion stability and bioavailability of the fat soluble active compared to emulsions stabilized with protein alone. Sodium alginate is a natural anionic linear polysaccharide extracted from brown algae or gulfweed, has thickening, emulsifying and gelling properties, and is widely applied to food, medicine and pharmaceutical industries. The sodium alginate has pH responsiveness, the molecular skeleton shrinks under the acidic environment, the pore diameter becomes small, the molecular structure expands under the neutral or alkaline environment, and the surface pore diameter becomes large, so that the release process of the loaded substance can be controlled, and the bioavailability of the loaded substance is improved. The corn starch is a processed product of corn, the surface of the corn starch is easy to generate physical and chemical modification, the mixing and dissolving of the corn starch between a water phase and an oil phase are facilitated, the stability of the emulsion is improved, and the corn starch serving as a stabilizer is widely applied to the field of food.
Disclosure of Invention
The invention provides a preparation method of a fish skin gelatin emulsion stabilized by sodium alginate and corn starch by taking cold water fish skin gelatin as an emulsifier. The invention uses fishskin gelatin as an emulsifier, sodium alginate and corn starch as stabilizers and corn oil as an oil phase carrier to obtain the fishskin gelatin emulsion which is stabilized by the sodium alginate and the corn starch and can embed fat-soluble active substances. Compared with the emulsion prepared by only using the fish skin gelatin under the same condition, the emulsion prepared by the invention has better stability, higher embedding rate of fat-soluble active substances and higher bioavailability.
The invention relates to a preparation method of a fish skin gelatin emulsion stabilized by sodium alginate and corn starch, which comprises the following steps:
s1, preparation of biopolymer stabilizer: mixing and dissolving sodium alginate and corn starch in deionized water, wherein the mass concentration of the sodium alginate is 0.125-0.25%, the mass concentration of the corn starch is 0.1-0.3%, performing water bath reaction at 85-95 ℃, and then cooling to room temperature to obtain a biopolymer stabilizer;
s2, preparing a water phase: dissolving the fish skin gelatin in the biopolymer stabilizing agent prepared in the step S1 to prepare a water phase;
s3, preparing an oil phase: weighing corn oil, and adding a fat-soluble active substance (such as benzyl isothiocyanate) serving as an oil phase, wherein the mass concentration of the fat-soluble active substance is 0-5 mg/mL; after the fat-soluble active substance is added into the corn oil, the fat-soluble active substance serving as an oil phase does not influence the stability of the emulsion of the product;
s4, mixing water phase and oil phase: mixing the oil phase prepared in the step S3 with the water phase prepared in the step S2, wherein the volume of the oil phase is 8-15% of that of the water phase, and then dispersing at a high speed;
s5, homogenizing: and homogenizing the product obtained in the step S4 under high pressure to obtain the fish skin gelatin emulsion stabilized by sodium alginate and corn starch.
Further, the water bath reaction time in the step S1 is 15-30 min;
further, in the biopolymer stabilizing agent of step S2, the mass concentration of fish skin gelatin is 1% to 3%;
further, in the step S4, the dispersion is carried out for 2-3 min under the condition of 10000-15000 r/min;
further, the oil phase volume of step S4 is 10% of the water phase, which is the most effective;
further, in step S5, the high-pressure homogenizing condition is 10000-12000 psi (psi: pounds per square foot) for 5-7 times.
The invention has the beneficial effects that:
the invention adopts fish-derived protein as an emulsifier, and can effectively solve a plurality of problems of infectious disease (mad cow disease) infection, religious belief and the like related to gelatin from other mammals. The sodium alginate and corn starch biopolymer can play respective roles as a stabilizer: the corn starch is mixed and dissolved in an oil-water interface, so that the stability of the emulsion is improved, the pH response characteristic of the sodium alginate inhibits the degradation of fat-soluble active substances embedded in the emulsion in the digestion process, and the bioavailability of the fat-soluble active substances is improved. The corn oil is used as an oil phase carrier, and the raw materials are easy to obtain, safe and convenient. The preparation method is simple and easy to implement, and compared with the pure fish skin gelatin, the prepared emulsion has high absolute value of electromotive potential and uniform distribution of the emulsion structure displayed by a cold field scanning electron microscope. Taking fat-soluble active substance Benzyl Isothiocyanate (BITC) as an example, compared with the emulsion prepared by only using fishskin gelatin under the same condition, the emulsion prepared by the invention is an oil-in-water conveying system which has the advantages of low cost, safety, convenience, higher BITC embedding rate and bioavailability and better storage stability, and has great potential in the development of products such as health food, functional beverage and the like.
The invention uses fish skin gelatin as an emulsifier and biological polymers of sodium alginate and corn starch as a stabilizer to obtain a stable emulsion; the stability of the emulsion, the embedding rate and the retention rate of fat-soluble active substances are improved compared with the pure fish skin gelatin emulsion, so that the product has higher efficacy value. The emulsion prepared by the invention has small average particle size and high absolute value of electromotive potential, and the cold field scanning electron microscope image shows that the emulsion has uniform spatial structure distribution and better stability, and can be used as an effective embedding system of fat-soluble active substances to be applied to the field of foods.
Drawings
FIG. 1 is a graph showing the particle size change of emulsions prepared in examples 1 to 4 of the present invention and emulsions prepared from a control group of pure fish skin gelatin when stored at 4 ℃ for 0 to 14 days;
FIG. 2 is a graph showing the potential change of emulsions prepared in examples 1 to 4 of the present invention and a control emulsion of pure fish skin gelatin (g) stored at 4 ℃ for 0 to 14 days;
FIG. 3 is a graph showing the appearance of emulsions prepared in examples 1 to 4 of the present invention and a control emulsion of simple fish skin gelatin (comparative example 1) stored at 4 ℃ for 0 day;
FIG. 4 shows the appearance of the product emulsions prepared in examples 1-4 of the present invention and the simple fish skin gelatin control emulsion (comparative example 1) stored at 4 ℃ for day 1;
FIG. 5 shows the appearance of the product emulsions prepared in examples 1-4 of the present invention and the simple fish skin gelatin control emulsion (comparative example 1) stored at 4 ℃ for 4 days;
FIG. 6 shows the appearance of the product emulsions prepared in examples 1-4 of the present invention and the simple fish skin gelatin control emulsion (comparative example 1) stored at 4 ℃ for 7 days;
FIG. 7 shows the appearance of the product emulsions prepared in examples 1-4 of the present invention and the pure fish skin gelatin control emulsion (comparative example 1) stored at 4 ℃ for 14 days;
FIG. 8 is a scanning electron micrograph of the inventive simple fish skin gelatin control emulsion at cold field (inset is a partial magnified view);
FIG. 9 is a cold field scanning electron micrograph (inset is enlarged partial view) of a product emulsion prepared according to example 1 of the present invention;
FIG. 10 is a cold field scanning electron micrograph (inset is enlarged partial view) of a product emulsion prepared according to example 2 of the present invention;
FIG. 11 is a cold field scanning electron micrograph (inset is enlarged partial view) of a product emulsion prepared according to example 3 of the present invention;
FIG. 12 is a cold field scanning electron micrograph (inset is a close-up view) of a product emulsion prepared according to example 4 of the present invention.
FIG. 13 is a graph of the BITC encapsulation efficiency of the product emulsion prepared in examples 5-8 of the present invention and a simple fish skin gelatin control emulsion;
FIG. 14 is a graph of the retention rate of BITC after simulated digestion of the product emulsions prepared in examples 5-8 of the present invention and the simple fish skin gelatin control emulsion.
Detailed Description
The test methods used in the following examples are conventional methods unless otherwise specified. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Comparative example 1: control group of simple fish skin gelatin
S1, weighing fish skin gelatin, dissolving the fish skin gelatin in deionized water, and preparing into an aqueous gelatin solution with the mass concentration of the fish skin gelatin being 1% for later use;
s2, oil phase preparation: weighing corn oil; dissolving benzyl isothiocyanate in corn oil, and sufficiently shaking to prepare an oil phase with the mass concentration of the benzyl isothiocyanate being 5mg/mL for later use;
s3, adding the oil phase obtained in the step S2 into the aqueous phase gelatin solution prepared in the step S1, wherein the volume of the oil phase is 10% of that of the aqueous phase gelatin solution, and then dispersing at a high speed of 10000r/min for 3 min;
s4, homogenizing the product of the step S3 for 6 times at 12000psi under high pressure to obtain the pure fish skin gelatin control emulsion.
Example 1
S1, mixing and dissolving sodium alginate and corn starch in deionized water, wherein the mass concentration of the sodium alginate is 0.125%, the mass concentration of the corn starch is 0.1%, heating to 90 ℃, carrying out water bath reaction for 20min, and then cooling to room temperature to obtain a stabilizer;
s2, weighing fish skin gelatin, dissolving the fish skin gelatin in the stabilizer prepared in the step S1, and preparing an aqueous phase gelatin solution with the mass concentration of the fish skin gelatin being 1% for later use;
s3, oil phase preparation: weighing corn oil as an oil phase;
s4, adding the oil phase obtained in the step S3 into the aqueous phase gelatin solution prepared in the step S2, wherein the volume of the oil phase is 10% of that of the aqueous phase gelatin solution, and then dispersing at a high speed of 12000r/min for 3 min;
s5, homogenizing the product of the step S4 for 5 times at 12000psi under high pressure to obtain the fish skin gelatin emulsion stabilized by sodium alginate and corn starch.
Example 2
S1, mixing and dissolving sodium alginate and corn starch in deionized water, wherein the mass concentration of the sodium alginate is 0.15%, the mass concentration of the corn starch is 0.2%, heating to 90 ℃, carrying out water bath reaction for 20min, and then cooling to room temperature to obtain a stabilizer;
s2, weighing fish skin gelatin, dissolving the fish skin gelatin in the stabilizer prepared in the step S1, and preparing an aqueous phase gelatin solution with the mass concentration of the fish skin gelatin being 1% for later use;
s3, oil phase preparation: weighing corn oil as an oil phase;
s4, adding the oil phase obtained in the step S3 into the aqueous phase gelatin solution prepared in the step S2, wherein the volume of the oil phase is 10% of that of the aqueous phase gelatin solution, and then dispersing at a high speed of 10000r/min for 3 min;
s5, homogenizing the product of the step S4 for 6 times at 12000psi under high pressure to obtain the fish skin gelatin emulsion stabilized by sodium alginate and corn starch.
Example 3
S1, mixing and dissolving sodium alginate and corn starch in deionized water, wherein the mass concentration of the sodium alginate is 0.2%, the mass concentration of the corn starch is 0.3%, heating to 90 ℃, carrying out water bath reaction for 20min, and then cooling to room temperature to obtain a stabilizer;
s2, weighing fish skin gelatin, dissolving the fish skin gelatin in the stabilizer prepared in the step S1, and preparing an aqueous phase gelatin solution with the mass concentration of the fish skin gelatin being 1% for later use;
s3, oil phase preparation: weighing corn oil as an oil phase;
s4, adding the oil phase obtained in the step S3 into the aqueous phase gelatin solution prepared in the step S2, wherein the volume of the oil phase is 10% of that of the aqueous phase gelatin solution, and then dispersing at a high speed of 15000r/min for 2 min;
s5, homogenizing the product of the step S4 for 5 times at 12000psi under high pressure to obtain the fish skin gelatin emulsion stabilized by sodium alginate and corn starch.
Example 4
S1, mixing and dissolving sodium alginate and corn starch in deionized water, wherein the mass concentration of the sodium alginate is 0.25%, the mass concentration of the corn starch is 0.1%, heating to 90 ℃, carrying out water bath reaction for 20min, and then cooling to room temperature to obtain a stabilizer;
s2, weighing fish skin gelatin, dissolving the fish skin gelatin in the stabilizer prepared in the step S1, and preparing an aqueous phase gelatin solution with the mass concentration of the fish skin gelatin being 1% for later use;
s3, oil phase preparation: weighing corn oil as an oil phase;
s4, adding the oil phase obtained in the step S3 into the aqueous phase gelatin solution prepared in the step S2, wherein the volume of the oil phase is 10% of that of the aqueous phase gelatin solution, and then dispersing at 13000r/min for 3min at a high speed;
s5, homogenizing the product of the step S4 for 7 times at 12000psi under high pressure to obtain the fish skin gelatin emulsion stabilized by sodium alginate and corn starch.
Example 5
S1, mixing and dissolving sodium alginate and corn starch in deionized water, wherein the mass concentration of the sodium alginate is 0.125%, the mass concentration of the corn starch is 0.1%, heating to 90 ℃, carrying out water bath reaction for 20min, and then cooling to room temperature to obtain a stabilizer;
s2, weighing fish skin gelatin, dissolving the fish skin gelatin in the stabilizer prepared in the step S1, and preparing an aqueous phase gelatin solution with the mass concentration of the fish skin gelatin being 1% for later use;
s3, oil phase preparation: weighing corn oil; dissolving benzyl isothiocyanate in corn oil, and sufficiently shaking to prepare an oil phase with the mass concentration of the benzyl isothiocyanate being 5mg/mL for later use;
s4, adding the oil phase obtained in the step S3 into the aqueous phase gelatin solution prepared in the step S2, wherein the volume of the oil phase is 10% of that of the aqueous phase gelatin solution, and then dispersing at high speed of 14000r/min for 2 min;
s5, homogenizing the product of the step S4 for 7 times at 12000psi under high pressure to obtain the fish skin gelatin emulsion stabilized by sodium alginate and corn starch.
Example 6
S1, mixing and dissolving sodium alginate and corn starch in deionized water, wherein the mass concentration of the sodium alginate is 0.15%, the mass concentration of the corn starch is 0.2%, heating to 90 ℃, carrying out water bath reaction for 20min, and then cooling to room temperature to obtain a stabilizer;
s2, weighing fish skin gelatin, dissolving the fish skin gelatin in the stabilizer prepared in the step S1, and preparing an aqueous phase gelatin solution with the mass concentration of the fish skin gelatin being 1% for later use;
s3, oil phase preparation: weighing corn oil; dissolving benzyl isothiocyanate in corn oil, and sufficiently shaking to prepare an oil phase with the mass concentration of the benzyl isothiocyanate being 5mg/mL for later use;
s4, adding the oil phase obtained in the step S3 into the aqueous phase gelatin solution prepared in the step S2, wherein the volume of the oil phase is 10% of that of the aqueous phase gelatin solution, and then dispersing at a high speed of 11000r/min for 3 min;
s5, homogenizing the product of the step S4 for 7 times at 12000psi under high pressure to obtain the fish skin gelatin emulsion stabilized by sodium alginate and corn starch.
Example 7
S1, mixing and dissolving sodium alginate and corn starch in deionized water, wherein the mass concentration of the sodium alginate is 0.2%, the mass concentration of the corn starch is 0.3%, heating to 90 ℃, carrying out water bath reaction for 20min, and then cooling to room temperature to obtain a stabilizer;
s2, weighing fish skin gelatin, dissolving the fish skin gelatin in the stabilizer prepared in the step S1, and preparing an aqueous phase gelatin solution with the mass concentration of the fish skin gelatin being 1% for later use;
s3, oil phase preparation: weighing corn oil; dissolving benzyl isothiocyanate in corn oil, and sufficiently shaking to prepare an oil phase with the mass concentration of the benzyl isothiocyanate being 5mg/mL for later use;
s4, adding the oil phase obtained in the step S3 into the aqueous phase gelatin solution prepared in the step S2, wherein the volume of the oil phase is 10% of that of the aqueous phase gelatin solution, and then dispersing at high speed of 14000r/min for 3 min;
s5, homogenizing the product of the step S4 for 7 times at 12000psi under high pressure to obtain the fish skin gelatin emulsion stabilized by sodium alginate and corn starch.
Example 8
S1, mixing and dissolving sodium alginate and corn starch in deionized water, wherein the mass concentration of the sodium alginate is 0.25%, the mass concentration of the corn starch is 0.1%, heating to 90 ℃, carrying out water bath reaction for 20min, and then cooling to room temperature to obtain a stabilizer;
s2, weighing fish skin gelatin, dissolving the fish skin gelatin in the stabilizer prepared in the step S1, and preparing an aqueous phase gelatin solution with the mass concentration of the fish skin gelatin being 1% for later use;
s3, oil phase preparation: weighing corn oil; dissolving benzyl isothiocyanate in corn oil, and sufficiently shaking to prepare an oil phase with the mass concentration of the benzyl isothiocyanate being 5mg/mL for later use;
s4, adding the oil phase obtained in the step S3 into the aqueous phase gelatin solution prepared in the step S2, wherein the volume of the oil phase is 10% of that of the aqueous phase gelatin solution, and then dispersing at a high speed of 12000r/min for 3 min;
s5, homogenizing the product of the step S4 for 7 times at 12000psi under high pressure to obtain the fish skin gelatin emulsion stabilized by sodium alginate and corn starch.
The particle size and the electromotive potential of the product emulsion prepared by the invention are measured: after diluting an appropriate amount of the product emulsions prepared in examples 1 to 4 (stored at 4 ℃ for 0 day, 1 day, 4 days, 7 days, and 14 days, respectively) by 100 times, the average particle size, the particle size distribution, and the electromotive potential were measured using a nano-particle size meter. Different letters in the data represent significant differences (p < 0.05). The concrete analysis and theoretical basis comprises:
(1) the average grain diameter of the emulsion is nano-grade, and the distribution is uniform and stable.
(2) The higher the absolute value of the zeta potential, the more stable the system.
The product emulsion prepared by the invention is measured by a cold field scanning electron microscope: after freezing the samples in liquid nitrogen, -80 ℃ sublimation for 20 min. And observing the microstructure of the product emulsion by adopting a cold field scanning electron microscope. The concrete analysis and theoretical basis comprises:
(1) the more uniform the emulsion structure in the cold field scanning electron microscope image, the more stable the system.
(2) The tighter the emulsion structure connection in the cold field scanning electron microscope image, the more stable the system.
Oil-in-water emulsion systems are frequently used for embedding fat-soluble active substances, and the product emulsion prepared by the invention is subjected to the measurement of the BITC embedding rate by taking BITC as an example: taking a proper amount of the product emulsion prepared in example 5 to example 8, extracting the embedded BITC in the emulsion with 1mL of normal hexane and 1mL of methanol, analyzing the content of the BITC by high performance liquid chromatography, and calculating the embedding rate (%) of the BITC in the emulsion according to a standard curve of the concentration of the BITC, wherein the embedding rate (%) is the peak area of the BITC in the emulsion/the peak area of the BITC in the standard substance.
Taking BITC as an example, the product emulsion prepared by the invention is subjected to BITC retention determination after being digested: an appropriate amount of the product emulsion prepared in example 5-example 8 was subjected to in vitro simulated digestion. The in vitro simulated digestion comprises the following specific steps: an in vitro simulated gastrointestinal tract (GIT) model is constructed, and the model consists of simulated digestive tracts of 3 stages of an oral cavity, a stomach and a small intestine so as to simulate the digestion process of the emulsion in vivo. The temperature of all solutions and dispersions was maintained at 37 ℃ throughout the digestion, and the pH of the samples was adjusted with 1M NaOH or HCl solution.
Simulating the oral digestion stage: mixing 20mL of emulsion sample preheated at 37 ℃ with 20mL of artificial saliva, adjusting the pH to 6.8, and shaking at constant temperature of 37 ℃ and 100rpm/min for 10min to simulate oral digestion;
simulating the gastric digestion stage: first, Simulated Gastric Fluid (SGF) was prepared by dissolving 2g NaCl and 7mL HCl in 1L distilled water. 20mL of the digested liquid from the mouth preheated to 37 ℃ was mixed with 20mL of SGF containing pepsin (3.2mg/mL) and the pH was adjusted to 2.5. Shaking at 37 deg.C and 100rpm/min for 2 hr to simulate gastric digestion;
simulating the small intestine digestion stage: first, 5.5g of CaCl was added2Simulated Intestinal Fluid (SIF) was prepared with 32.9g NaCl in 1L distilled water. 30mL of the digested liquid was placed in a 100mL beaker and stirred in a constant temperature heating magnetic stirrer at 37 ℃ at 100rpm/min with constant stirring and pH adjusted to 7.0. To the sample was added 1.5mL of SIF and 3.5mL of bile salt (53.6mg/mL), and the pH was adjusted to 7.0 again. Finally, 2.5mL of lipase solution (24mg/mL) was added to the system, and the pH of the system was maintained at 7.0 by adding 0.02M NaOH solution dropwise to the sample over 2h to simulate small intestine digestion.
Taking 1mL of digestive juice after the small intestine digestion stage, extracting the BITC in the emulsion by using 1mL of normal hexane and 1mL of methanol, measuring the content of the BITC by adopting high performance liquid chromatography, and calculating the retention rate (%) of the BITC in the emulsion according to a standard curve of the concentration of the BITC, wherein the retention rate (%) is the peak area of the BITC in the small intestine digestive juice/the peak area of the BITC in the standard substance.
The results of the tests are shown in fig. 1 to 14, and different letters in the data represent significant differences (p < 0.05).
In FIG. 1, all samples did not change much in particle size during 14 days of storage, and the particle size of the fish skin gelatin control group alone was slightly larger than that of the example group.
In fig. 2, all samples showed little potential change during 14 days of storage, and the absolute values of the potentials were greater in the example group than in the skin gelatin control group alone.
In FIGS. 3 to 7, all samples were stable and free from flocculation during 14 days of storage.
And FIGS. 8 to 12 are cold field scanning electron micrographs of a control group and examples 1 to 4 in sequence, and the sodium alginate filaments can be clearly observed to be interwoven on the fish skin gelatin branch structure along with the increase of the sodium alginate concentration, so that the stability of the system is increased.
FIG. 13 shows the results of the entrapment efficiency of all samples, when the concentration of sodium alginate is 0.25%, the entrapment efficiency is up to 95.45%, which is much higher than 68.88% of the control group. Compared with a pure fish skin gelatin control group, the embedding rate of the example group is obviously improved, which shows that the addition of sodium alginate and corn starch improves the encapsulation rate of the emulsion.
FIG. 14 shows the results of bioavailability of all samples, with the highest entrapment rate of 90.49% at a sodium alginate concentration of 0.25%, which is much higher than 80.12% of the control group. Illustrating the better protection of the embedded fat-soluble active substance during digestion in the examples of the present invention.
Compared with the pure fish skin gelatin emulsion, the emulsion prepared by the invention has higher absolute value of electromotive potential and uniform and compact structure of the emulsion in a cold field scanning electron microscope image. The slightly increased particle size indicates that the sodium alginate and the fish skin gelatin are fully compounded at an oil-water interface and play a role in protection in the digestion process of an emulsion system. Compared with the pure fish skin gelatin emulsion, the embedding rate and the bioavailability of the BITC are higher, namely the fish skin gelatin emulsion with stable sodium alginate and corn starch prepared by the invention can be used as a conveying system to improve the utilization rate of an embedded object. The sodium alginate and corn starch stabilized fish skin gelatin emulsion prepared in example 4 has very high stability.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be able to cover the technical solutions and the inventive concepts of the present invention within the technical scope of the present invention.
Claims (8)
1. A preparation method of a fish skin gelatin emulsion stabilized by sodium alginate and corn starch comprises the following steps:
s1, preparation of biopolymer stabilizer: mixing and dissolving sodium alginate and corn starch in deionized water, wherein the mass concentration of the sodium alginate is 0.125-0.25%, the mass concentration of the corn starch is 0.1-0.3%, performing water bath reaction at 85-95 ℃, and then cooling to room temperature to obtain a biopolymer stabilizer;
s2, preparing a water phase: dissolving the fish skin gelatin in the biopolymer stabilizing agent prepared in the step S1 to prepare a water phase;
s3, preparing an oil phase: weighing corn oil, adding fat-soluble active substances into the corn oil as an oil phase, wherein the mass concentration of the fat-soluble active substances is 0-5 mg/mL; after the fat-soluble active substance is added into the corn oil, the fat-soluble active substance serving as an oil phase does not influence the stability of the emulsion of the product;
s4, mixing water phase and oil phase: mixing the oil phase prepared in the step S3 with the water phase prepared in the step S2, wherein the volume of the oil phase is 8-15% of that of the water phase, and then dispersing at a high speed;
s5, homogenizing: and (5) homogenizing the product obtained in the step S4 under high pressure to obtain the fish skin gelatin emulsion stabilized by sodium alginate and corn starch.
2. The method for preparing a sodium alginate and corn starch stabilized fish skin gelatin emulsion as claimed in claim 1, wherein: and step S1, the water bath reaction time is 15-30 min.
3. The method for preparing a sodium alginate and corn starch stabilized fish skin gelatin emulsion as claimed in claim 1, wherein: in the biopolymer stabilizing agent of step S2, the mass concentration of fish skin gelatin is 1% to 3%.
4. The method for preparing a sodium alginate and corn starch stabilized fish skin gelatin emulsion as claimed in claim 1, wherein: the fat-soluble active substance in step S3 is benzyl isothiocyanate.
5. The method for preparing a sodium alginate and corn starch stabilized fish skin gelatin emulsion as claimed in claim 1, wherein: in the step S4, the high-speed dispersion condition is 10000-15000 r/min for 2-3 min.
6. The method for preparing a sodium alginate and corn starch stabilized fish skin gelatin emulsion as claimed in claim 1, wherein: the volume of the oil phase in step S4 was 10% of the water phase.
7. The method for preparing a sodium alginate and corn starch stabilized fish skin gelatin emulsion as claimed in claim 1, wherein: in step S5, the high-pressure homogenization is performed for 5-7 times under 10000-12000 psi.
8. A sodium alginate and corn starch stabilized fish skin gelatin emulsion is characterized in that: is prepared by the method of any one of claims 1 to 7.
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