CN111357971A - 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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- CN111357971A CN111357971A CN202010327250.1A CN202010327250A CN111357971A CN 111357971 A CN111357971 A CN 111357971A CN 202010327250 A CN202010327250 A CN 202010327250A CN 111357971 A CN111357971 A CN 111357971A
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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
- A23L29/015—Inorganic compounds
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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, belonging to the field of modern health food processing. The invention provides a preparation method of a double-crosslinked pickering emulsion, which comprises the steps of preparing pickering emulsion of gliadin-sodium alginate composite particles and preparing pickering emulsion of calcium ion and glutamine transaminase crosslinking, and comprises the following steps: preparing gliadin-curcumin mixed solution; preparing a sodium alginate stock solution; preparing a gliadin suspension loaded with curcumin; and (3) obtaining pickering emulsion of gliadin-sodium alginate composite particles or pickering emulsion of calcium ion and glutamine transaminase crosslinking. The invention discloses a method for constructing curcumin-loaded composite particles by using gliadin and sodium alginate for the first time to form stable pickering emulsion which can be used for embedding food nutrient substances.
Description
Technical Field
The invention relates to a double-crosslinked pickering emulsion and a preparation method and application thereof, belonging to the field of modern health food processing.
Background
At present, due to unreasonable diet caused by rapid life style, the phenomenon that the whole social population is in a sub-health state is caused, and the public is widely concerned. The concern of people on health is promoted to the demand of health food, however, most active substances with health care effects, such as curcumin, polyphenol and the like have the disadvantages of poor water solubility, instability and the like, so that a carrier is often required to be constructed for carrying, and the disadvantages are solved. In order to meet the requirements of people, the construction of food-grade transportation systems with different scales, such as nano particles, emulsions and the like, has great significance.
At present, Pickering emulsion is a hot point of research, but since Pickering (Pickering) emulsion is a thermodynamically unstable system, the structure of Pickering emulsion is often damaged in the presence of shear force such as freeze drying. Therefore, it is important to enhance the interface technology in research.
The wheat gluten is natural protein extracted from wheat (flour), is light yellow, has protein content as high as 75-85%, contains 15 kinds of amino acids essential for human body, is one kind of plant protein source with rich nutrients and low cost, and is extracted from wheat gluten. Sodium alginate is a natural linear anionic polysaccharide, has strong negative charges in a wide pH range (3-7), and is widely applied to food and chemical industries because of excellent thickening property, stability and safety.
At present, Pickering emulsion prepared by domestic and overseas researches is a thermodynamically unstable system, and high fluidity of the emulsion can increase collision among liquid drops to cause instability of liquid drop fusion emulsion. The freeze-drying stability of the emulsion is therefore limited, which affects its use in commercial food products. By combining the current research situations at home and abroad, no relevant report on the preparation of stable pickering emulsion of gliadin-sodium alginate complex by a double-crosslinking mode is found.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a preparation method of a double-crosslinked pickering emulsion.
The preparation method of the double-crosslinked pickering emulsion comprises a preparation method of pickering emulsion of Gliadin-sodium alginate (Gliadin-SA) composite particles and a preparation method of pickering emulsion of calcium ion and glutamine transaminase crosslinking.
In order to achieve the purpose, the invention adopts the technical scheme that: a preparation method of a double-crosslinked Pickering emulsion comprises the following steps:
(1) preparation of Gliadin (Gliadin) -curcumin (curcumin) mixed solution: weighing Gliadin in a container, adding ethanol to obtain Gliadin solution, adding curcumin (curcumin), stirring until the solution is clear, and hydrating at low temperature to obtain Gliadin-curcumin (Gliadin-Cur) mixed solution;
(2) preparation of sodium alginate (sodium alginate) stock solution: weighing a Sodium Alginate (SA) sample, dissolving the Sodium Alginate (SA) sample in a phosphate buffer solution to obtain a sodium alginate (sodium alginate) stock solution, and refrigerating for storage;
(3) preparation of gliadin suspension loaded with curcumin: 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 suspension loaded with curcumin;
(4) preparation of pickering emulsion of Gliadin-sodium alginate (sodium alginate) composite particles: adding peanut oil into the Gliadin suspension loaded with curcumin in the step (3), centrifuging and homogenizing, adding the sodium alginate stock solution in the step (2), and adjusting the pH value to obtain stable pickering emulsion of Gliadin-sodium alginate (Gliadin-SA) particles;
or calcium ion (Ca)2+) Preparation of pickering emulsion crosslinked with glutamine transaminase (TGase): adding glutamine transaminase into the gliadin suspension loaded with curcumin in the step (3), reacting in a constant-temperature water bath, then inactivating enzyme at high temperature, then adding peanut oil, centrifuging and homogenizing, then adding calcium chloride and the sodium alginate (sodium alginate) storage solution in the step (2), adjusting the concentration of the calcium chloride, and adjusting the pH value to obtain calcium ions (Ca)2+) Pickering emulsion crosslinked with a transglutaminase (TGase).
At present, the preparation technology of the stable Pickering emulsion of the compound reported in the prior art is seriously insufficient, the novel method for preparing the Pickering emulsion by using the double cross-linking agent provided by the invention can be used for embedding food nutrient substances in the future.
In the preparation method of the double-crosslinked pickering emulsion, the gliadin is extracted from the 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 and divalent cations in water can generate a gelling effect to form microspheres with a controlled release effect, and the microspheres can be used for conveying nutrient substances.
As a preferred embodiment of the preparation method of the double-crosslinked pickering emulsion, in the step (1), the gliadin accounts for 4 parts by weight, the volume concentration of the ethanol is 65-80%, the curcumin accounts for 0.02 part by weight, 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 preferred embodiment of the preparation method of the double-crosslinked pickering emulsion of the invention, in the step (2), 0.1 weight part of the double-crosslinked pickering emulsion is dissolved in 100mL of 0.05M phosphate buffer solution with pH of 7.0 to prepare 0.1% (w/v) sodium alginate (sodium alginate) stock solution, and the temperature for cold storage is 1-6 ℃.
As a preferred embodiment of the preparation method of the double-crosslinked pickering emulsion, 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 operation of removing ethanol is rotary evaporation at 50-60 ℃.
As a preferred embodiment of the method for preparing a double-crosslinked pickering emulsion according to the present invention, in the step (4), the preparation of the pickering emulsion of the gliadin-sodium alginate composite particles: the volume ratio of the gliadin suspension 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 stock solution to the gliadin suspension loaded with curcumin is 1-0.9-1.1, and the pH value is adjusted to be 3.5-7.0 by NaOH.
As a preferred embodiment of the method for preparing the double-crosslinked pickering emulsion of the present invention, the concentration of NaOH is 1M, and the pH 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 in which calcium ions are crosslinked with glutamine transaminase is prepared: the volume part of the gliadin suspension loaded with curcumin is 1 part, the specific activity of glutamine transaminase is 30U/g, the temperature of the 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 ℃, the treatment time is 10-20 min, the volume ratio of the gliadin suspension 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 the centrifugal homogenization is 1-3 min, the volume ratio of the gliadin suspension loaded with curcumin to the sodium alginate liquid storage 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), the temperature of the calcium ion and the transglutaminase for crosslinking is 50 ℃, the time of crosslinking is 1h, the crosslinking sequence is that the transglutaminase crosslinks gliadin to prepare the emulsion, and Ca is added after the emulsion is formed2+
As a preferred embodiment of the method for preparing the double-crosslinked pickering emulsion of the present invention, the pH is 3.5, and the concentration of NaOH is 1M.
Another object of the present invention is a double-crosslinked pickering emulsion obtained with the method for preparing a double-crosslinked pickering emulsion according to the invention.
It is a further object of the present invention to provide the use of said double cross-linked 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, gliadin and sodium alginate are utilized to construct curcumin-carried composite particles for the first time to form stable pickering emulsion which can be used for embedding food nutrient substances;
(2) the Pickering emulsion prepared by the preparation method of the double-crosslinked Pickering emulsion has pH responsiveness;
(3) the invention utilizes the stable Pickering emulsion of Gliadin-sodium alginate (Gliadin-SA) particles which are rich in calcium ions and glutamine transaminase to carry out cross-linking treatment to obtain calcium ions (Ca)2+) Pickering emulsions crosslinked with transglutaminase (TGase) provide improved hardness, cohesion, tack and recovery properties relative to Ca2+The improvement of the Pickering emulsion crosslinked with TGase alone is more obvious, which shows that the Pickering emulsion when the calcium ions and the glutamine transaminase are doubly crosslinked has more excellent oxidation resistance compared with the crosslinking of a single component while the texture characteristic is obviously improved.
Drawings
FIG. 1 is a graph showing the results of rheological measurements of stable Pickering emulsions of Gliadin-sodium alginate (Gliadin-SA) composite particles obtained in example 1 of the present invention using an MCR92 rheometer under different pH (3.5 or, 5.5, 7.0) systems;
FIG. 2 is a graph showing the results of the oxidation stability test of Pickering emulsions prepared in example 2 of the present invention, comparative example 1 and comparative example 2 using a gas chromatograph-Agilent 7890B.
Detailed Description
To better illustrate the objects, aspects and advantages of the present invention, the present invention will be further described with reference to specific examples.
Example 1
The embodiment is a preparation method of a double-crosslinked pickering emulsion, which is a preparation method of a pickering emulsion of gliadin-sodium alginate composite particles, and includes the following steps:
(1) preparation of Gliadin (Gliadin) -curcumin (curcumin) mixed solution: weighing 4g of Gliadin, adding 70% (v/v) ethanol to prepare Gliadin solution, adding 0.02g of Curcumin (Curcumin), magnetically stirring until the solution is clear, and hydrating at 4 ℃ for more than 8h to obtain Gliadin-Curcumin (Gliadin-Cur) mixed solution;
(2) preparation of sodium alginate (sodium alginate) stock solution: weighing 0.1g of Sodium Alginate (SA) sample, dissolving in 100ml of 0.05M phosphate buffer solution with pH of 7.0 to prepare 0.1% (w/v) sodium alginate stock solution, and refrigerating for storage;
(3) preparation of gliadin suspension loaded with curcumin: quickly pouring deionized water with the volume ratio of 1:7 into the Gliadin-curcumin (Gliadin-Cur) mixed solution obtained in the step (1) under the stirring state, and then performing rotary evaporation at 55 ℃ to remove ethanol to obtain a Gliadin suspension loaded with curcumin;
(4) preparation of pickering emulsion of gliadin-sodium alginate composite particles: and (3) taking 10mL of the Gliadin suspension loaded with curcumin in the step (3), adding 8.8mL of peanut oil, centrifuging at 20000rpm/min, homogenizing for 2min, then adding 10mL of 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 stable pickering emulsion of Gliadin-sodium alginate (Gliadin-SA) composite particles.
Example 2
This example illustrates the preparation of a double-crosslinked pickering emulsion of the present invention, which is calcium ion (Ca)2+) A method for preparing a pickering emulsion crosslinked with a transglutaminase (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 Gliadin solution, adding 0.02g of Curcumin (Curcumin), magnetically stirring until the solution is clear, and hydrating at 4 ℃ for more than 8h to obtain Gliadin-Curcumin (Gliadin-Cur) mixed solution;
(2) preparation of sodium alginate (sodium alginate) stock solution: weighing 0.1g of Sodium Alginate (SA) sample, dissolving in 100ml of 0.05M phosphate buffer solution with pH of 7.0 to prepare 0.1% (w/v) sodium alginate stock solution, and refrigerating at 4 ℃;
(3) preparation of gliadin suspension loaded with curcumin: quickly pouring deionized water with the volume ratio of 1:7 into the Gliadin-curcumin (Gliadin-Cur) mixed solution obtained in the step (1) under the stirring state, and then performing rotary evaporation at 55 ℃ to remove ethanol to obtain a Gliadin suspension loaded with curcumin;
(4) preparation of a pickering emulsion in which calcium ions are crosslinked with glutamine transaminase: taking 1 part by volume of the gliadin suspension loaded with curcumin in the step (3), adding 30U/g of glutamine transaminase, reacting in a constant-temperature water bath at 50 ℃ for 60min, treating at 75 ℃ for 15min to inactivate enzyme, then adding 8.8mL of peanut oil, centrifuging at 20000rpm/min to homogenize for 2min, then adding 10mL of calcium chloride and 10mL of sodium alginate stock solution in the step (2) to ensure that the concentration of calcium chloride in the system is 3.5% (w/v), and adjusting the pH to 3.5 or 5.5 or 7.0 by using 1MNaOH to obtain calcium ions (Ca2+) Pickering emulsion crosslinked with a transglutaminase (TGase).
Example 3
The embodiment is a preparation method of a double-crosslinked pickering emulsion, which is a preparation method of a pickering emulsion of gliadin-sodium alginate composite particles, and includes the following steps:
(1) preparation of Gliadin (Gliadin) -curcumin (curcumin) mixed solution: weighing 4g of Gliadin, adding 65% (v/v) ethanol to prepare Gliadin solution, adding 0.02g of Curcumin (Curcumin), magnetically stirring until the solution is clear, and hydrating at 1 ℃ for more than 8h to obtain Gliadin-Curcumin (Gliadin-Cur) mixed solution;
(2) preparation of sodium alginate (sodium alginate) stock solution: weighing 0.1g of Sodium Alginate (SA) sample, dissolving in 100ml of 0.05M phosphate buffer solution with pH of 7.0 to prepare 0.1% (w/v) sodium alginate stock solution, and refrigerating at 1 deg.C;
(3) preparation of gliadin suspension loaded with curcumin: quickly pouring deionized water with a volume ratio of 1:6 into the Gliadin-curcumin (Gliadin-Cur) mixed solution obtained in the step (1) under a stirring state, and then performing rotary evaporation at 50 ℃ to remove ethanol to obtain a Gliadin suspension loaded with curcumin;
(4) preparation of pickering emulsion of gliadin-sodium alginate composite particles: and (3) taking 10mL of the Gliadin suspension loaded with curcumin in the step (3), adding 8mL of peanut oil, centrifuging at 20000rpm/min, homogenizing for 2min, then adding 9mL of 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 stable pickering emulsion of 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 a pickering emulsion of gliadin-sodium alginate composite particles, and includes the following steps:
(1) preparation of Gliadin (Gliadin) -curcumin (curcumin) mixed solution: weighing 4g of Gliadin, adding 80% (v/v) ethanol to prepare Gliadin solution, adding 0.02g of Curcumin (Curcumin), magnetically stirring until the solution is clear, and hydrating at 6 ℃ for more than 8h to obtain Gliadin-Curcumin (Gliadin-Cur) mixed solution;
(2) preparation of sodium alginate (sodium alginate) stock solution: weighing 0.1g of Sodium Alginate (SA) sample, dissolving in 100ml of 0.05M phosphate buffer solution with pH of 7.0 to prepare 0.1% (w/v) sodium alginate stock solution, and refrigerating at 6 ℃;
(3) preparation of gliadin suspension loaded with curcumin: quickly pouring deionized water with a volume ratio of 1:8 into the Gliadin-curcumin (Gliadin-Cur) mixed solution obtained in the step (1) under a stirring state, and then performing rotary evaporation at 60 ℃ to remove ethanol to obtain a Gliadin suspension loaded with curcumin;
(4) preparation of pickering emulsion of gliadin-sodium alginate composite particles: and (3) taking 10mL of the Gliadin suspension loaded with curcumin in the step (3), adding 11mL of peanut oil, centrifuging at 20000rpm/min and homogenizing for 2min, then adding 11mL of sodium alginate stock solution in the step (2), and adjusting the pH value to 3.5 or 5.5 or 7.0 by using 1M NaOH to obtain the stable pickering emulsion of Gliadin-sodium alginate (Gliadin-SA) composite particles.
Example 5
This example illustrates the preparation of a double-crosslinked pickering emulsion of the present invention, which is calcium ion (Ca)2+) A method for preparing a pickering emulsion crosslinked with a transglutaminase (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 Gliadin solution, adding 0.02g of Curcumin (Curcumin), magnetically stirring until the solution is clear, and hydrating at 1 ℃ for more than 8h to obtain Gliadin-Curcumin (Gliadin-Cur) mixed solution;
(2) preparation of sodium alginate (sodium alginate) stock solution: weighing 0.1g of Sodium Alginate (SA) sample, dissolving in 100ml of 0.05M phosphate buffer solution with pH of 7.0 to prepare 0.1% (w/v) sodium alginate stock solution, and refrigerating at 1 deg.C;
(3) preparation of gliadin suspension loaded with curcumin: quickly pouring deionized water with a volume ratio of 1:6 into the Gliadin-curcumin (Gliadin-Cur) mixed solution obtained in the step (1) under a stirring state, and then performing rotary evaporation at 50 ℃ to remove ethanol to obtain a Gliadin suspension loaded with curcumin;
(4) preparation of a pickering emulsion in which calcium ions are crosslinked with glutamine transaminase: taking 1 part by volume of the gliadin suspension loaded with curcumin in the step (3), adding 30U/g of glutamine transaminase, reacting in a constant-temperature water bath at 45 ℃ for 50min, treating at 75 ℃ for 10min to inactivate enzyme, then adding 8mL of peanut oil, centrifuging at 18000rpm/min to homogenize for 2min, then adding 9mL of calcium chloride and the sodium alginate stock solution in the step (2) to make the concentration of the calcium chloride in the system be 3%, and adjusting the pH to 3.5 or 5.5 or 7.0 by using 1M NaOH to obtain calcium ions (Ca)2+) Pickering emulsion crosslinked with a transglutaminase (TGase).
Example 6
This example illustrates the preparation of a double-crosslinked pickering emulsion of the present invention, which is calcium ion (Ca)2+) A method for preparing a pickering emulsion crosslinked with a transglutaminase (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 Gliadin solution, adding 0.02g of Curcumin (Curcumin), magnetically stirring until the solution is clear, and hydrating at 6 ℃ for more than 8h to obtain Gliadin-Curcumin (Gliadin-Cur) mixed solution;
(2) preparation of sodium alginate (sodium alginate) stock solution: weighing 0.1g of Sodium Alginate (SA) sample, dissolving in 100ml of 0.05M phosphate buffer solution with pH of 7.0 to prepare 0.1% (w/v) sodium alginate stock solution, and refrigerating at 6 ℃;
(3) preparation of gliadin suspension loaded with curcumin: quickly pouring deionized water with a volume ratio of 1:8 into the Gliadin-curcumin (Gliadin-Cur) mixed solution obtained in the step (1) under a stirring state, and then performing rotary evaporation at 60 ℃ to remove ethanol to obtain a Gliadin suspension loaded with curcumin;
(4) preparation of a pickering emulsion in which calcium ions are crosslinked with glutamine transaminase: taking 1 part by volume of the gliadin suspension loaded with curcumin in the step (3), adding 30U/g of glutamine transaminase, reacting in a constant-temperature water bath at 55 ℃ for 70min, treating at 75 ℃ for 20min to inactivate enzyme, then adding 11mL of peanut oil, centrifuging at 22000rpm/min to homogenize for 2min, then adding 11mL of calcium chloride and the sodium alginate solution in the step (2) to enable the concentration of the calcium chloride in the system to be 5% (w/v), and adjusting the pH to 3.5 or 5.5 or 7.0 by using 1M NaOH to obtain calcium ions (Ca is calcium ions)2+) Pickering emulsion crosslinked with a transglutaminase (TGase).
Comparative example 1
This comparative example is a process for the preparation of a transglutaminase (TGase) crosslinked pickering emulsion comprising the steps of:
(1) preparation of Gliadin (Gliadin) -curcumin (curcumin) mixed liquor: weighing 4g of Gliadin, adding 70% (v/v) ethanol to prepare Gliadin solution, adding 0.02g of Curcumin (Curcumin), magnetically stirring until the solution is clear, and hydrating at 4 ℃ for more than 8h to obtain Gliadin-Cur mixed solution;
(2) preparation of sodium alginate (sodium alginate) stock solution: weighing 0.1gSA sample, dissolving in 100mL0.05MpH7.0 phosphate buffer solution, preparing 0.1% (w/v) sodium alginate (sodium alginate) stock solution, and refrigerating;
(3) quickly pouring the Gliadin-Cur mixed solution in the step (1) into deionized water with a volume ratio of 1:7 under magnetic stirring, and removing ethanol by rotary evaporation at 55 ℃ to obtain a Gliadin suspension loaded with curcumin;
(4) and (3) taking 10mL of the Gliadin suspension loaded with curcumin in the step (3), adding 30U/g of TGase, reacting in a constant-temperature water bath at 50 ℃ for 1h, treating at 75 ℃ for 15 minutes to inactivate enzyme, then adding 8.8mL of peanut oil, homogenizing at 20000rpm/min for 2min, adding 10mL of the sodium alginate stock solution in the step (2), and adjusting the pH to 3.5 by using 1M NaOH to obtain the glutamine transaminase cross-linked Pickering emulsion.
Comparative example 2
This comparative example is calcium ion (Ca)2+) A method of preparing a crosslinked pickering emulsion comprising the steps of:
(1) preparation of Gliadin (Gliadin) -curcumin (curcumin) mixed liquor: weighing 4g of Gliadin, adding 70% (v/v) ethanol to prepare Gliadin solution, adding 0.02g of Curcumin (Curcumin), magnetically stirring until the solution is clear, and hydrating at 4 ℃ for more than 8h to obtain Gliadin-Cur mixed solution;
(2) preparation of sodium alginate (sodium alginate) stock solution: weighing 0.1gSA sample, dissolving in 100mL0.05MpH7.0 phosphate buffer solution, preparing 0.1% (w/v) sodium alginate (sodium alginate) stock solution, and refrigerating;
(3) quickly pouring the Gliadin-Cur mixed solution in the step (1) into deionized water with a volume ratio of 1:7 under magnetic stirring, and removing ethanol by rotary evaporation at 55 ℃ to obtain a Gliadin suspension loaded with curcumin;
(4) and (3) taking 10mL of Gliadin suspension with curcumin loaded in the suspension, adding 8.8mL of peanut oil, homogenizing at 20000rpm/min for 2min, adding 10mL of sodium alginate stock solution and calcium chloride in the step (2) to enable the concentration of the calcium chloride in the system to be 3.5%, and adjusting the pH value to be 3.5 by using 1MNaOH to obtain the calcium ion crosslinked Pickering emulsion.
Experimental example 1
In this experimental example, an MCR92 rheometer was used to perform rheological measurements on the stable pickering emulsion of Gliadin-sodium alginate (Gliadin-SA) composite particles obtained in example 1 of the present invention under different pH (3.5 or, 5.5, 7.0) systems, and the experimental results are shown in fig. 1.
The experimental results are as follows:
fig. 1A can see that, under the amplitude strain sweep, the elasticity modulus (G') and the viscosity modulus (G ") of the stable pickering emulsion of the Gliadin-sodium alginate (Gliadin-SA) composite particles did not change much with increasing shear stress, indicating that the internal structure of the stable pickering emulsion of the Gliadin-sodium alginate (Gliadin-SA) composite particles of example 1 was not damaged; after the shear stress is increased continuously, the two curves are crossed, the corresponding shear stress at the intersection point is called yield stress, at the moment, the structure of the stable pickering emulsion of 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 increase of the pH value.
Figure 1B all Gliadin-sodium alginate (Gliadin-SA) composite particle stabilized pickering emulsions at pH3.5 or 5.5 or 7.0 under amplitude frequency sweep consistently have greater elastic modulus (G') than viscous modulus (G ") under corresponding stress, 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.
Figure 1C shows a decrease in apparent viscosity as shear rate increases, illustrating that the stable pickering emulsion of the Gliadin-sodium alginate (Gliadin-SA) composite particles of example 1 is a non-newtonian fluid.
Taken together, it can be determined from the above results that adjusting the pH can change the rheological properties exhibited by the stable pickering emulsion of 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 of the present invention were tested for oxidation stability by using a gas chromatograph-Agilent 7890B, and the mass-structure characteristics of the pickering emulsions prepared were measured by using a mass-structure analyzer after vacuum freeze-drying, and the experimental results are shown in fig. 2. The results of comparing the 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 results of the detection of different Pickering emulsion indexes
As can be seen from the data results of Table 1, comparative example 2 calcium ion (Ca)2+) The crosslinked pickering emulsion has improved four indexes of hardness (N), cohesive force (-), viscosity (-) and recoverability (-); comparative example 1 the effect of the crosslinked pickering emulsion of transglutaminase (TGase) on four indexes of hardness (N), cohesion (-), tack (N) and recovery (-) is more significant than that of comparative example 2; while the pickering emulsion of example 2 is calcium ion (Ca)2+) When the emulsion is simultaneously crosslinked with glutamine transaminase (TGase), the improvement of four index characteristics of hardness (N), cohesive force (-), viscosity (N) and recovery property (-) of the 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, Ca alone is used in the system of pH3.52+The storage stability of the system is slightly reduced when the cross-linking is effected with transglutaminase (TG enzyme) alone, but when Ca is used2+The decrease in the content of n-hexane, a secondary oxidation product produced by accelerated oxidation upon double-crosslinking with TG enzyme, indicates that Ca of example 2 of the present invention2+The Pickering emulsion prepared by double cross-linking with TG enzyme has remarkably improved texture characteristics and excellent oxidation resistance.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
Claims (10)
1. A preparation method of a double-crosslinked Pickering emulsion is characterized by comprising the following steps:
(1) preparing a gliadin-curcumin mixed solution: weighing gliadin in a container, adding ethanol to prepare gliadin solution, adding curcumin, stirring until the solution is clear, and hydrating at low temperature to obtain gliadin-curcumin mixed solution;
(2) preparation of sodium alginate stock solution: weighing a sodium alginate sample, dissolving the sodium alginate sample in a phosphate buffer solution to obtain a sodium alginate stock solution, and refrigerating for storage;
(3) preparation of gliadin suspension loaded with curcumin: taking the gliadin-curcumin mixed solution in the step (1), pouring deionized water into the mixed solution under a stirring state, and then removing ethanol to obtain a gliadin suspension loaded with curcumin;
(4) preparation of pickering emulsion of gliadin-sodium alginate composite particles: adding peanut oil into the gliadin suspension loaded with curcumin in the step (3), centrifuging and homogenizing, adding the sodium alginate stock solution in the step (2), and adjusting the pH value to obtain a pickering emulsion of gliadin-sodium alginate composite particles;
or the preparation of a pickering emulsion in which calcium ions are crosslinked with glutamine transaminase: and (3) adding glutamine transaminase into the gliadin suspension loaded with curcumin in the step (3), carrying out constant-temperature water bath reaction, then carrying out high-temperature enzyme deactivation, then adding peanut oil, carrying out centrifugal homogenization, then 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 crosslinked by calcium ions and the glutamine transaminase.
2. The method for preparing the double-crosslinked pickering emulsion according to claim 1, wherein in the step (1), the gliadin is 4 parts by weight, the volume concentration of the ethanol is 65-80%, the curcumin is 0.02 part by weight, 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 the double-crosslinked pickering emulsion as claimed in claim 1, wherein in the step (2), 0.1 weight part of the double-crosslinked pickering emulsion is dissolved in 100mL of 0.05M phosphate buffer solution with pH of 7.0 to prepare 0.1% (w/v) sodium alginate stock solution, and the temperature for cold storage is 1-6 ℃.
4. The method for preparing the 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 a double-crosslinked pickering emulsion according to claim 1, wherein in step (4), the preparation of the pickering emulsion of the gliadin-sodium alginate composite particles: the volume ratio of the gliadin suspension 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 loaded with curcumin to the sodium alginate stock solution is 1: 0.9-1.1, and the pH value is adjusted to be 3.5-7.0 by NaOH.
6. The method of preparing a double-crosslinked pickering emulsion of claim 5, wherein the NaOH is present at a concentration of 1M and the pH is 3.5, 5.5, or 7.0.
7. The method for preparing a double-crosslinked pickering emulsion according to claim 1, wherein in the step (4), the pickering emulsion in which calcium ions are crosslinked with glutamine transaminase is prepared: the volume part of the gliadin suspension loaded with curcumin is 1 part, the specific activity of glutamine transaminase is 30U/g, the temperature of the 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 carried out at 75 ℃ for 10-20 min, the volume ratio of the gliadin suspension 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 the centrifugal homogenization is 1-3 min, the volume ratio of the gliadin suspension loaded with curcumin to the sodium alginate liquid storage 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 of 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 for producing a double-crosslinked pickering emulsion according to any one of claims 1 to 8.
10. Use of the double-crosslinked pickering emulsion of claim 9 in food processing.
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