CN111357971B - Double-crosslinked Pickering emulsion and preparation method and application thereof - Google Patents
Double-crosslinked Pickering emulsion and preparation method and application thereof Download PDFInfo
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- CN111357971B CN111357971B CN202010327250.1A CN202010327250A CN111357971B CN 111357971 B CN111357971 B CN 111357971B CN 202010327250 A CN202010327250 A CN 202010327250A CN 111357971 B CN111357971 B CN 111357971B
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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
- 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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Abstract
The invention relates to a double-crosslinked Pickering emulsion, and a preparation method and application thereof, and belongs to the field of modern health food processing. The preparation method of the double-crosslinked pickering emulsion provided by the invention comprises the following steps of pickering emulsion of gliadin-sodium alginate composite particles and pickering emulsion crosslinked by calcium ions and glutamine transaminase: preparing a gliadin-curcumin mixed solution; preparing sodium alginate stock solution; preparing a gliadin suspension loaded with curcumin; and (3) obtaining the pickering emulsion of the gliadin-sodium alginate composite particles or the pickering emulsion of the cross-linked calcium ions and glutamine transaminase. The invention discloses that the gliadin and sodium alginate are utilized to construct composite particles carrying curcumin for the first time to form stable Pickering emulsion, which can be used for embedding food nutrients.
Description
Technical Field
The invention relates to a double-crosslinked Pickering emulsion, and a preparation method and application thereof, and belongs to the field of modern health food processing.
Background
At present, the phenomenon that the whole social crowd presents sub-health state is caused by unreasonable diet caused by a rapid life style, and the phenomenon is widely paid attention to. The interest of health has been raised to the demand for health foods, however, most active substances with health care effects, such as curcumin and polyphenol, have the disadvantages of poor water solubility, instability and the like, so that carrier mounting is often required to be constructed, and the disadvantages are solved. In order to meet the demands of people, the construction of food-grade transportation systems with different scales such as nano particles, emulsion and the like is of great significance.
At present, pickering emulsions are the hot spot of research, but because Pickering (Pickering) emulsions are thermodynamically unstable systems, the Pickering (Pickering) emulsions tend to break their structures in the presence of shear forces such as freeze drying. Therefore, enhancing the interface technology becomes a serious issue in research.
The wheat gluten is natural protein extracted from wheat (flour) and has light yellow color, protein content up to 75% -85%, and 15 amino acids essential to human body, and is a plant protein source with rich nutrients and low quality. Sodium alginate is a natural linear anionic polysaccharide, has strong negative charges in a wide pH range (3-7), and is widely applied to the food and chemical industries due to excellent thickening property, stability and safety.
At present, pickering emulsion prepared by research at home and abroad is a thermodynamically unstable system, and the high fluidity of the emulsion can increase collision among liquid drops, so that the liquid drop fusion emulsion is unstable. Thus limiting the freeze-drying stability of the emulsion, which would affect its use in commercial foods. The research current situation at home and abroad is integrated, and no relevant report on the preparation of the stable pickering emulsion of the gliadin-sodium alginate complex by a double-crosslinking mode is seen.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a preparation method of double-crosslinked pickering emulsion.
The preparation method of the double-crosslinked pickering emulsion provided by the invention comprises the preparation method of pickering emulsion of Gliadin-sodium alginate (Gliadin-SA) composite particles and the preparation method of pickering emulsion crosslinked by calcium ions and glutamine transaminase.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows: the preparation method of the double-crosslinked Pickering emulsion comprises the following steps:
(1) Preparation of Gliadin (Gliadin) -curcumin (curcumin) mixed solution: weighing Gliadin, adding ethanol into a container to prepare Gliadin solution, adding curcumin (curcumin), stirring until the solution is clear, and performing low-temperature hydrolysis to obtain Gliadin-curcumin (Gliadin-Cur) mixed solution;
(2) Preparation of sodium alginate (sodium alginate) stock solution: weighing Sodium Alginate (SA) sample, dissolving in phosphate buffer solution to obtain sodium alginate (sodium alginate) stock solution, and refrigerating;
(3) Preparation of a curcumin-loaded gliadin suspension: pouring deionized water into the Gliadin-curcumin mixed solution (Gliadin-Cur) in the step (1) in a stirring state, and then removing ethanol to obtain a Gliadin-loaded Gliadin suspension;
(4) Preparation of Pickering emulsion of Gliadin (Gliadin) -sodium alginate (sodium alginate) composite particles: taking the Gliadin suspension liquid loaded with curcumin in the step (3), adding peanut oil, centrifuging and homogenizing, adding the sodium alginate stock solution in the step (2), and adjusting the pH value to obtain a Gliadin-sodium alginate (Gliadin-SA) particle-stabilized pickering emulsion;
or calcium ion (Ca) 2+ ) Preparation of pickering emulsion crosslinked with glutamine transaminase (TGase): taking the gliadin suspension carrying curcumin in the step (3), adding glutamine transaminase, carrying out a constant-temperature water bath reaction, inactivating enzyme at a high temperature, adding peanut oil, centrifuging and homogenizing, adding calcium chloride and sodium alginate (sodium alginate) stock solution in the step (2), regulating the concentration of the calcium chloride, and regulating the pH value to obtain calcium ions (Ca) 2+ ) Pickering emulsions crosslinked with glutamine transaminase (TGase).
At present, the reported preparation technology of the pickering emulsion with stable compound is seriously insufficient, and the novel method for preparing the pickering emulsion by using the double cross-linking agent can be used for embedding food nutrients in future.
In the preparation method of the double-crosslinked pickering emulsion, the gliadin is extracted from wheat gluten, and has high hydrophobicity due to the high hydrophobic amino acid content of the gliadin, and becomes a good choice for stabilizing the pickering emulsion due to the high safety of the gliadin.
In the preparation method of the double-crosslinked pickering emulsion, sodium alginate can perform a gel action with divalent cations in water to form microspheres with a controlled release effect, and the microspheres can be used for conveying nutrient substances.
In the step (1), in the preferred implementation mode of the preparation method of the double-crosslinked pickering emulsion, in the step (1), the weight portion of gliadin is 4, the volume concentration of ethanol is 65% -80%, the weight portion of curcumin is 0.02, the stirring is magnetic stirring, the low-temperature hydration temperature is 1-6 ℃, and the low-temperature hydration time is more than 8 hours.
In the step (2), 0.1 parts by weight of the double crosslinked pickering emulsion is dissolved in 100mL of a phosphate buffer solution with a pH of 7.0 at 0.05M to prepare a sodium alginate (sodium alginate) stock solution with a concentration of 0.1% (w/v), and the temperature of the cold storage is 1-6 ℃.
In the step (3), the volume ratio of the Gliadin-curcumin mixed solution (Gliadin-Cur) to the deionized water is 1:6-8, the stirring is magnetic stirring, and the ethanol removal operation is rotary evaporation at 50-60 ℃.
As a preferred embodiment of the preparation method of the double-crosslinked pickering emulsion, in the step (4), the pickering emulsion of the gliadin-sodium alginate composite particle is prepared by: the volume ratio of the gliadin suspension liquid loaded with curcumin to the peanut oil is 1:0.8-1.1, the speed of centrifugal homogenization is 18000-22000 rpm/min, the time of centrifugal homogenization is 1-3 min, the volume ratio of the sodium alginate liquid storage to the gliadin suspension liquid loaded with curcumin is 1-0.9-1.1, and the pH value is regulated to 3.5-7.0 by NaOH.
As a preferred embodiment of the preparation method of the double-crosslinked pickering emulsion, the concentration of NaOH is 1M, and the pH value is 3.5, 5.5 or 7.0.
As a preferred embodiment of the method for preparing a double-crosslinked pickering emulsion according to the present invention, in the step (4), a pickering emulsion crosslinked by calcium ions and glutamine transaminase is prepared: the volume fraction of the gliadin-loaded gliadin suspension is 1 part, the specific activity of glutamine transaminase is 30U/g, the temperature of a constant-temperature water bath is 45-55 ℃, the time of the constant-temperature water bath is 50-70 min, the high-temperature enzyme deactivation is 75 ℃ for 10-20 min, the volume ratio of the gliadin-loaded gliadin suspension to peanut oil is 1:0.8-1.1, the speed of centrifugation homogenization is 18000-22000 rpm/min, the time of centrifugation homogenization is 1-3 min, the volume ratio of the gliadin-loaded gliadin suspension to the stock solution of sodium alginate is 1:0.9-1.1, the mass concentration of calcium chloride is 3.0-5.0%, and the pH value is adjusted to 3.5 or 5.5 or 7.0 by NaOH
As a preferred embodiment of the preparation method of the double-crosslinked pickering emulsion, in the step (4), calcium ions and glutamine transaminase are crosslinked at 50 ℃ for 1h, and the crosslinked order is that the gliadin is crosslinked by the glutamine transaminase to prepare emulsion, and Ca is added after the emulsion is formed 2+
As a preferred embodiment of the method for preparing a double crosslinked pickering emulsion according to the present invention, the pH is 3.5, and the NaOH concentration is 1M.
The invention also aims at obtaining the double-crosslinked Pickering emulsion by adopting the preparation method of the double-crosslinked Pickering emulsion.
It is a further object of the present invention to provide the use of the double crosslinked pickering emulsion in food processing.
Compared with the prior art, the invention has the beneficial effects that:
(1) In the preparation method of the double-crosslinked pickering emulsion, composite particles carrying curcumin are constructed by using gliadin and sodium alginate for the first time, so that stable pickering emulsion is formed, and the pickering emulsion can be used for embedding food nutrients;
(2) The Pickering emulsion prepared by the preparation method of the double-crosslinked Pickering emulsion has pH responsiveness;
(3) The invention uses the stable Pickering emulsion of Gliadin-sodium alginate (Gliadin-SA) particles with more calcium ions and glutamine transaminase to carry out crosslinking treatment, and the obtained calcium ions (Ca) 2+ ) Pickering emulsions crosslinked with glutamine transaminase (TGase) have improved hardness, cohesion, adhesion and recovery properties to the Pickering emulsionFor Ca 2+ The pickering emulsion crosslinked with TGase alone is more remarkable in improvement, which shows that the pickering emulsion crosslinked with calcium ions and glutamine transaminase has more excellent oxidation resistance compared with the single-component crosslinking while the texture characteristics are remarkably improved.
Drawings
FIG. 1 is a graph showing the results of rheological measurements of a Pickering emulsion stabilized with a MCR92 rheometer on Gliadin-sodium alginate (Gliadin-SA) composite particles obtained in example 1 of the present invention at different pH (3.5 or 5.5 or 7.0) systems;
FIG. 2 is a graph showing the results of the test of oxidation stability of pickering emulsions prepared in example 2, comparative example 1 and comparative example 2 according to the present invention using a gas chromatograph-Agilent 7890B.
Detailed Description
For a better description of the objects, technical solutions and advantages of the present invention, the present invention will be further described with reference to the following specific examples.
Example 1
The embodiment is a preparation method of a double-crosslinked pickering emulsion, which is a preparation method of pickering emulsion of gliadin-sodium alginate composite particles, and comprises the following steps:
(1) Preparation of Gliadin (Gliadin) -curcumin (curcumin) mixed solution: weighing 4g of Gliadin, adding 70% (v/v) ethanol to prepare a Gliadin solution, adding 0.02g of Curcumin (Curcumin), magnetically stirring until the solution is clear, and hydrating at 4 ℃ for more than 8 hours to obtain a Gliadin-Curcumin (Gliadin-Cur) mixed solution;
(2) Preparation of sodium alginate (sodium alginate) stock solution: weighing 0.1g Sodium Alginate (SA) sample, dissolving in 100ml0.05M phosphate buffer solution with pH7.0, preparing 0.1% (w/v) sodium alginate stock solution, and refrigerating for preservation;
(3) Preparation of a curcumin-loaded gliadin suspension: taking the Gliadin-curcumin (Gliadin-Cur) mixed solution in the step (1), rapidly pouring deionized water with a volume ratio of 1:7 in a stirring state, and then removing ethanol by rotary evaporation at 55 ℃ to obtain a Gliadin-loaded Gliadin suspension;
(4) Preparation of Pickering emulsion of gliadin-sodium alginate composite particles: taking 10mL of the Gliadin-loaded Gliadin suspension in the step (3), adding 8.8mL of peanut oil, centrifuging and homogenizing for 2min at 20000rpm/min, adding 10mL of the sodium alginate stock solution in the step (2), and adjusting the pH to 3.5, 5.5 or 7.0 by using 1M NaOH to obtain the Gliadin-sodium alginate (Gliadin-SA) composite particle stable pickering emulsion.
Example 2
The embodiment is a preparation method of the double-crosslinked pickering emulsion, wherein the preparation method of the double-crosslinked pickering emulsion is calcium ion (Ca) 2+ ) A process for the preparation of a pickering emulsion crosslinked with a glutamine transaminase (TGase), comprising the steps of:
(1) Preparation of Gliadin (Gliadin) -curcumin (curcumin) mixed solution: weighing 4g of Gliadin, adding 70% (v/v) ethanol to prepare a Gliadin solution, adding 0.02g of Curcumin (Curcumin), magnetically stirring until the solution is clear, and hydrating at 4 ℃ for more than 8 hours to obtain a Gliadin-Curcumin (Gliadin-Cur) mixed solution;
(2) Preparation of sodium alginate (sodium alginate) stock solution: weighing 0.1g Sodium Alginate (SA) sample, dissolving in 100ml0.05M phosphate buffer solution with pH7.0, preparing 0.1% (w/v) sodium alginate stock solution, and refrigerating at 4deg.C;
(3) Preparation of a curcumin-loaded gliadin suspension: taking the Gliadin-curcumin (Gliadin-Cur) mixed solution in the step (1), rapidly pouring deionized water with a volume ratio of 1:7 in a stirring state, and then removing ethanol by rotary evaporation at 55 ℃ to obtain a Gliadin-loaded Gliadin suspension;
(4) Preparation of pickering emulsion crosslinked by calcium ions and glutamine transaminase: taking 1 part of the gliadin suspension carrying curcumin in the step (3), adding 30U/g of glutamine transaminase, reacting for 60min in a constant-temperature water bath at 50 ℃, treating for 15min at 75 ℃ to inactivate enzymes, and then adding 8.8mL of peanut oil and 200Centrifuging at 00rpm/min, homogenizing for 2min, adding calcium chloride and 10mL of the sodium alginate stock solution obtained in step (2) to make the calcium chloride concentration in the system 3.5% (w/v), and adjusting pH to 3.5 or 5.5 or 7.0 with 1M NaOH to obtain calcium ion (Ca) 2+ ) Pickering emulsions crosslinked with glutamine transaminase (TGase).
Example 3
The embodiment is a preparation method of a double-crosslinked pickering emulsion, which is a preparation method of pickering emulsion of gliadin-sodium alginate composite particles, and comprises the following steps:
(1) Preparation of Gliadin (Gliadin) -curcumin (curcumin) mixed solution: weighing 4g of Gliadin, adding 65% (v/v) ethanol to prepare a Gliadin solution, adding 0.02g of Curcumin (Curcumin), magnetically stirring until the solution is clear, and hydrating at 1 ℃ for more than 8 hours to obtain a Gliadin-Curcumin (Gliadin-Cur) mixed solution;
(2) Preparation of sodium alginate (sodium alginate) stock solution: weighing 0.1g Sodium Alginate (SA) sample, dissolving in 100ml0.05M phosphate buffer solution with pH7.0, preparing 0.1% (w/v) sodium alginate stock solution, and refrigerating at 1 deg.C;
(3) Preparation of a curcumin-loaded gliadin suspension: taking the Gliadin-curcumin (Gliadin-Cur) mixed solution in the step (1), rapidly pouring deionized water with a volume ratio of 1:6 in a stirring state, and then removing ethanol by rotary evaporation at 50 ℃ to obtain a Gliadin-loaded Gliadin suspension;
(4) Preparation of Pickering emulsion of gliadin-sodium alginate composite particles: taking 10mL of the Gliadin suspension liquid loaded with curcumin in the step (3), adding 8mL of peanut oil, centrifuging and homogenizing for 2min at 20000rpm, adding 9mL of the sodium alginate stock solution in the step (2), and adjusting the pH to 3.5 or 5.5 or 7.0 by using 1M NaOH to obtain the Pickering emulsion with stable Gliadin-sodium alginate (Gliadin-SA) composite particles.
Example 4
The embodiment is a preparation method of a double-crosslinked pickering emulsion, which is a preparation method of pickering emulsion of gliadin-sodium alginate composite particles, and comprises the following steps:
(1) Preparation of Gliadin (Gliadin) -curcumin (curcumin) mixed solution: weighing 4g of Gliadin, adding 80% (v/v) ethanol to prepare a Gliadin solution, adding 0.02g of Curcumin (Curcumin), magnetically stirring until the solution is clear, and hydrating at 6 ℃ for more than 8 hours to obtain a Gliadin-Curcumin (Gliadin-Cur) mixed solution;
(2) Preparation of sodium alginate (sodium alginate) stock solution: weighing 0.1g Sodium Alginate (SA) sample, dissolving in 100ml0.05M phosphate buffer solution with pH7.0, preparing 0.1% (w/v) sodium alginate stock solution, and refrigerating at 6deg.C;
(3) Preparation of a curcumin-loaded gliadin suspension: taking the Gliadin-curcumin (Gliadin-Cur) mixed solution in the step (1), rapidly pouring deionized water with a volume ratio of 1:8 in a stirring state, and then removing ethanol by rotary evaporation at 60 ℃ to obtain a Gliadin-loaded Gliadin suspension;
(4) Preparation of Pickering emulsion of gliadin-sodium alginate composite particles: taking 10mL of the Gliadin suspension liquid loaded with curcumin in the step (3), adding 11mL of peanut oil, centrifuging and homogenizing for 2min at 20000rpm, adding 11mL of the sodium alginate stock solution in the step (2), and adjusting the pH to 3.5 or 5.5 or 7.0 by using 1M NaOH to obtain the Gliadin-sodium alginate (Gliadin-SA) composite particle stable Pickering emulsion.
Example 5
The embodiment is a preparation method of the double-crosslinked pickering emulsion, wherein the preparation method of the double-crosslinked pickering emulsion is calcium ion (Ca) 2+ ) A process for the preparation of a pickering emulsion crosslinked with a glutamine transaminase (TGase), comprising the steps of:
(1) Preparation of Gliadin (Gliadin) -curcumin (curcumin) mixed solution: weighing 4g of Gliadin, adding 65% (v/v) ethanol to prepare a Gliadin solution, adding 0.02g of Curcumin (Curcumin), magnetically stirring until the solution is clear, and hydrating at 1 ℃ for more than 8 hours to obtain a Gliadin-Curcumin (Gliadin-Cur) mixed solution;
(2) Preparation of sodium alginate (sodium alginate) stock solution: weighing 0.1g Sodium Alginate (SA) sample, dissolving in 100ml0.05M phosphate buffer solution with pH7.0, preparing 0.1% (w/v) sodium alginate stock solution, and refrigerating at 1 deg.C;
(3) Preparation of a curcumin-loaded gliadin suspension: taking the Gliadin-curcumin (Gliadin-Cur) mixed solution in the step (1), rapidly pouring deionized water with a volume ratio of 1:6 in a stirring state, and then removing ethanol by rotary evaporation at 50 ℃ to obtain a Gliadin-loaded Gliadin suspension;
(4) Preparation of pickering emulsion crosslinked by calcium ions and glutamine transaminase: taking 1 part by volume of the gliadin suspension carrying curcumin in the step (3), adding 30U/g glutamine transaminase, reacting for 50min in a constant-temperature water bath at 45 ℃, then treating for 10min to inactivate enzyme, then adding 8mL peanut oil, centrifuging at 18000rpm/min to homogenize for 2min, then adding calcium chloride and 9mL sodium alginate stock solution in the step (2), so that the concentration of calcium chloride in the system is 3%, and adjusting the pH value to 3.5 or 5.5 or 7.0 by using 1M NaOH to obtain calcium ions (Ca 2+ ) Pickering emulsions crosslinked with glutamine transaminase (TGase).
Example 6
The embodiment is a preparation method of the double-crosslinked pickering emulsion, wherein the preparation method of the double-crosslinked pickering emulsion is calcium ion (Ca) 2+ ) A process for the preparation of a pickering emulsion crosslinked with a glutamine transaminase (TGase), comprising the steps of:
(1) Preparation of Gliadin (Gliadin) -curcumin (curcumin) mixed solution: weighing 4g of Gliadin, adding 80% (v/v) ethanol to prepare a Gliadin solution, adding 0.02g of Curcumin (Curcumin), magnetically stirring until the solution is clear, and hydrating at 6 ℃ for more than 8 hours to obtain a Gliadin-Curcumin (Gliadin-Cur) mixed solution;
(2) Preparation of sodium alginate (sodium alginate) stock solution: weighing 0.1g Sodium Alginate (SA) sample, dissolving in 100ml0.05M phosphate buffer solution with pH7.0, preparing 0.1% (w/v) sodium alginate stock solution, and refrigerating at 6deg.C;
(3) Preparation of a curcumin-loaded gliadin suspension: taking the Gliadin-curcumin (Gliadin-Cur) mixed solution in the step (1), rapidly pouring deionized water with a volume ratio of 1:8 in a stirring state, and then removing ethanol by rotary evaporation at 60 ℃ to obtain a Gliadin-loaded Gliadin suspension;
(4) Preparation of pickering emulsion crosslinked by calcium ions and glutamine transaminase: taking 1 part of gliadin suspension carrying curcumin in the step (3), adding 30U/g glutamine transaminase, reacting for 70min in a constant-temperature water bath at 55 ℃, then treating for 20min to inactivate enzyme, then adding 11mL peanut oil, centrifuging at 22000rpm/min to homogenize for 2min, then adding calcium chloride and 11mL sodium alginate stock solution in the step (2), so that the concentration of calcium chloride in the system is 5% (w/v), and regulating the pH value to 3.5 or 5.5 or 7.0 by using 1M NaOH to obtain calcium ions (Ca) 2+ ) Pickering emulsions crosslinked with glutamine transaminase (TGase).
Comparative example 1
The comparative example is a process for the preparation of a crosslinked pickering emulsion of glutamine transaminase (TGase), comprising the steps of:
(1) Preparation of Gliadin (Gliadin) -curcumin (curcumin) mixture: weighing 4g of Gliadin, adding 70% (v/v) ethanol to prepare a Gliadin solution, adding 0.02g of Curcumin (Curcumin), magnetically stirring until the solution is clear, and hydrating at 4 ℃ for more than 8 hours to obtain a Gliadin-Cur mixed solution;
(2) Preparation of sodium alginate (sodium alginate) stock solution: weighing 0.1gSA sample, dissolving in 100ml0.05M phosphate buffer solution with pH7.0, preparing into 0.1% (w/v) sodium alginate stock solution, and refrigerating for preservation;
(3) Taking the Gliadin-Cur mixed solution in the step (1), rapidly pouring deionized water with a volume ratio of 1:7 under magnetic stirring, and removing ethanol by rotary evaporation at 55 ℃ to obtain a curcumin-loaded Gliadin suspension;
(4) Taking 10mL of the curcumin-loaded Gliadin suspension in the step (3), adding 30U/g of TGase, carrying out a constant-temperature water bath reaction at 50 ℃ for 1h, then treating at 75 ℃ for 15 minutes to inactivate enzymes, then adding 8.8mL of peanut oil, homogenizing at 20000rpm/min for 2min, then adding 10mL of the sodium alginate stock solution in the step (2), and regulating the pH to 3.5 by using 1M NaOH to obtain the crosslinked pickering emulsion of the glutamine transaminase.
Comparative example 2
The comparative example is calcium ion (Ca 2+ ) A method for preparing a crosslinked pickering emulsion comprising the steps of:
(1) Preparation of Gliadin (Gliadin) -curcumin (curcumin) mixture: weighing 4g of Gliadin, adding 70% (v/v) ethanol to prepare a Gliadin solution, adding 0.02g of Curcumin (Curcumin), magnetically stirring until the solution is clear, and hydrating at 4 ℃ for more than 8 hours to obtain a Gliadin-Cur mixed solution;
(2) Preparation of sodium alginate (sodium alginate) stock solution: weighing 0.1gSA sample, dissolving in 100ml0.05M phosphate buffer solution with pH7.0, preparing into 0.1% (w/v) sodium alginate stock solution, and refrigerating for preservation;
(3) Taking the Gliadin-Cur mixed solution in the step (1), rapidly pouring deionized water with a volume ratio of 1:7 under magnetic stirring, and removing ethanol by rotary evaporation at 55 ℃ to obtain a curcumin-loaded Gliadin suspension;
(4) Taking 10mL of suspension to load curcumin Gliadin suspension, adding 8.8mL of peanut oil, homogenizing for 2min at 20000rpm/min, adding 10mL of sodium alginate stock solution and calcium chloride in the step (2), enabling the concentration of calcium chloride in the system to be 3.5%, and regulating the pH value to 3.5 by using 1M NaOH to obtain the calcium ion crosslinked Pickering emulsion.
Experimental example 1
The experimental example adopts an MCR92 rheometer to carry out rheological measurement on the pickering emulsion with stable Gliadin-sodium alginate (Gliadin-SA) composite particles obtained in the invention example 1 under different pH (3.5 or 5.5 or 7.0) systems, and the experimental result is shown in figure 1.
Experimental results:
as can be seen from fig. 1A, under the amplitude strain scan, the elastic modulus (G') and the viscous modulus (G ") of the pickering emulsion stabilized by the Gliadin-sodium alginate (Gliadin-SA) composite particles have a small tendency to change with the increase of the shear stress, which indicates that the internal structure of the pickering emulsion stabilized by the Gliadin-sodium alginate (Gliadin-SA) composite particles of example 1 is not destroyed; after the shear stress is continuously increased, the two curves are crossed, the shear stress corresponding to the intersection point is called yield stress, at the moment, the structure of the Pickering emulsion stabilized by the Gliadin-sodium alginate (Gliadin-SA) composite particles is changed, and the yield stress of the Pickering emulsion is reduced from 280Pa to 21Pa along with the rising of pH.
FIG. 1B shows that all Gliadin-sodium alginate (Gliadin-SA) composite particle-stabilized pickering emulsions at pH3.5 or 5.5 or 7.0 have an elastic modulus (G') always greater than the viscous modulus (G ") at the corresponding stress under the sweep of the amplitude frequency, indicating that the Gliadin-sodium alginate (Gliadin-SA) composite particle-stabilized pickering emulsion of example 1 has colloidal properties at pH3.5 or 5.5 or 7.0.
FIG. 1C shows a decrease in apparent viscosity as shear rate increases, illustrating that the Gliadin-sodium alginate (Gliadin-SA) composite particle-stabilized pickering emulsion of example 1 is a non-Newtonian fluid.
From the above results, it was determined that adjusting the pH can change the rheological properties exhibited by the pickering emulsion stabilized by the Gliadin-sodium alginate (Gliadin-SA) composite particles of example 1.
Experimental example 2
The pickering emulsions prepared in example 2, comparative example 1 and comparative example 2 were subjected to oxidation stability test by using a gas chromatograph-Agilent 7890B, and the texture characteristics of the prepared pickering emulsions were measured by using a texture analyzer after vacuum freeze-drying, and the experimental results are shown in fig. 2. The results of comparison of four indexes of hardness (N), cohesion (-), tackiness (N) and recovery (-) of the pickering emulsions prepared in example 2, comparative example 1 and comparative example 2 are shown in Table 1.
TABLE 1 detection results of different Pickering emulsion indexes
From the data results of Table 1As can be seen, comparative example 2 calcium ion (Ca 2+ ) The crosslinked Pickering emulsion has improved four indexes of hardness (N), cohesive force (-), viscosity (-) and resilience (-); whereas the effect of comparative example 1 on four indicators of hardness (N), cohesion (-), adhesion (N) and recovery (-) was more pronounced for the pick lin emulsion crosslinked with glutamine transaminase (TGase) than for comparative example 2; whereas the pickering emulsion of example 2 was calcium (Ca 2+ ) When simultaneously crosslinked with glutamine transaminase (TGase), the improvement of four index characteristics of hardness (N), cohesion (-), viscosity (N) and recovery (-) of pickering emulsion is higher and more remarkable than that of comparative example 2 and comparative example 1; the different letters (a-d) in Table 1 indicate that there is a significant difference (P<0.05)。
As can be seen from FIG. 2, at pH3.5, ca alone 2+ The storage stability of the system is slightly reduced when the crosslinking is carried out with glutamine transaminase alone (TG enzyme), but when Ca 2+ The decrease in n-hexane content of the secondary oxidation product produced by accelerated oxidation upon double crosslinking with TG enzyme indicates Ca according to example 2 of the present invention 2+ The pickering emulsion prepared by double cross-linking with the TG enzyme has obviously improved texture characteristics and excellent oxidation resistance.
Finally, it should be noted that the above embodiments are only for illustrating the technical solution of the present invention and not for limiting the scope of the present invention, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solution of the present invention may be modified or substituted equally without departing from the spirit and scope of the technical solution of the present invention.
Claims (10)
1. The preparation method of the double-crosslinked Pickering emulsion is characterized by comprising the following steps of:
(1) Preparation of gliadin-curcumin mixed solution: weighing gliadin in a container, adding ethanol with the volume concentration of 65-80% to prepare a gliadin solution, adding curcumin, stirring until the solution is clear, and carrying out low-temperature hydrolysis to obtain a gliadin-curcumin mixed solution;
(2) Preparation of sodium alginate stock solution: weighing sodium alginate samples, dissolving in phosphate buffer solution to obtain sodium alginate stock solution, and refrigerating for preservation;
(3) Preparation of a curcumin-loaded gliadin suspension: taking the gliadin-curcumin mixed solution in the step (1), pouring deionized water under a stirring state, and then removing ethanol to obtain a gliadin-loaded gliadin suspension;
(4) Preparation of pickering emulsion crosslinked by calcium ions and glutamine transaminase: taking the gliadin suspension carrying curcumin in the step (3), adding glutamine transaminase, carrying out a constant-temperature water bath reaction, inactivating enzyme at a high temperature, adding peanut oil, centrifuging and homogenizing, adding calcium chloride and the sodium alginate stock solution in the step (2), adjusting the concentration of the calcium chloride, and adjusting the pH value to obtain the pickering emulsion with calcium ions and glutamine transaminase crosslinked.
2. The method for preparing a double-crosslinked pickering emulsion according to claim 1, wherein in step (1), the weight portion of gliadin is 4 portions, the weight portion of curcumin is 0.02 portion, the stirring is magnetic stirring, the low-temperature hydration temperature is 1-6 ℃, and the low-temperature hydration time is more than 8 hours.
3. The method for preparing a double-crosslinked pickering emulsion according to claim 1, wherein in step (2), 0.1 parts by weight of the double-crosslinked pickering emulsion is dissolved in 100ml of phosphate buffer solution with a pH of 7.0 at 0.05M to prepare a sodium alginate stock solution with a concentration of 0.1% (w/v), and the temperature of cold storage is 1-6 ℃.
4. The method for preparing a double-crosslinked pickering emulsion according to claim 1, wherein in the step (3), the volume ratio of the gliadin-curcumin mixed solution to the deionized water is 1:6-8, the stirring is magnetic stirring, and the ethanol removal operation is rotary evaporation at 50-60 ℃.
5. The method for preparing the double-crosslinked pickering emulsion according to claim 1, wherein in the step (4), the pickering emulsion of the gliadin-sodium alginate composite particles is prepared by: the volume ratio of the gliadin suspension liquid loaded with curcumin to the peanut oil is 1:0.8-1.1, the speed of centrifugal homogenization is 18000-22000 rpm/min, the time of centrifugal homogenization is 1-3 min, the volume ratio of the gliadin suspension liquid loaded with curcumin to the sodium alginate stock solution is 1:0.9-1.1, and the pH value is regulated to 3.5-7.0 by NaOH.
6. The method for preparing the double-crosslinked pickering emulsion according to claim 5, wherein the concentration of NaOH is 1M, and the pH value is 3.5, 5.5 or 7.0.
7. The method for preparing a double crosslinked pickering emulsion according to claim 1, wherein in step (4), calcium ion and glutamine transaminase crosslinked pickering emulsion is prepared: the volume fraction of the gliadin-loaded gliadin suspension is 1 part, the specific activity of glutamine transaminase is 30U/g, the temperature of a constant-temperature water bath is 45-55 ℃, the time of the constant-temperature water bath is 50-70 min, the high-temperature enzyme deactivation is 75 ℃ for 10-20 min, the volume ratio of the gliadin-loaded gliadin suspension to peanut oil is 1:0.8-1.1, the speed of centrifugation homogenization is 18000-22000 rpm/min, the time of centrifugation homogenization is 1-3 min, the volume ratio of the gliadin-loaded gliadin suspension to the stock solution of sodium alginate is 1:0.9-1.1, the mass concentration of calcium chloride is 3.0-5.0%, and the pH value is adjusted to 3.5, 5.5 or 7.0 by NaOH.
8. The method for preparing a double crosslinked pickering emulsion of claim 7, wherein the pH is 3.5 and the NaOH concentration is 1M.
9. A double crosslinked pickering emulsion obtained by the method of preparing a double crosslinked pickering emulsion of any one of claims 1-8.
10. Use of the double crosslinked pickering emulsion of claim 9 in food processing.
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| CN115944074A (en) * | 2022-12-12 | 2023-04-11 | 武汉轻工大学 | Preparation method of Pickering emulsion, embedding and delivering method of bioactive substances and 3D printing method of food |
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