CN115024493A - Active ingredient loaded nutrition microsphere and preparation method and application thereof - Google Patents

Active ingredient loaded nutrition microsphere and preparation method and application thereof Download PDF

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Publication number
CN115024493A
CN115024493A CN202210713518.4A CN202210713518A CN115024493A CN 115024493 A CN115024493 A CN 115024493A CN 202210713518 A CN202210713518 A CN 202210713518A CN 115024493 A CN115024493 A CN 115024493A
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microspheres
oil
protein
pickering emulsion
nutritional
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谭明乾
王丽丽
刘辰玥
庄英男
苏文涛
程沙沙
王海涛
相思源
宋玉昆
于德洋
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Asia Fishing Port Ltd By Share Ltd
Dalian Polytechnic University
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Asia Fishing Port Ltd By Share Ltd
Dalian Polytechnic University
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    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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    • A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
    • A23J1/14—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds
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    • 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
    • A23L29/03—Organic compounds
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    • 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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    • 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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    • A23L29/10—Foods or foodstuffs containing additives; Preparation or treatment thereof containing emulsifiers
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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    • A23P10/30—Encapsulation of particles, e.g. foodstuff additives
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    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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Abstract

The invention discloses active ingredient loaded nutrient microspheres and a preparation method and application thereof, belonging to the field of food. The invention adopts natural edible protein aqueous solution as water phase, grease as oil phase, and natural edible protein as superfine particle emulsifier at the same time to prepare Pickering emulsion; then, by introducing grape seed extract, green tea extract, polysaccharide, sodium alginate and calcium ions, the microspheres generate protein-protein and alginic acid-calcium double cross-linking, and the stable millimeter-grade nutritional microspheres are obtained. According to the invention, the Pickering emulsion and the nutritional microspheres are combined, so that the double protection effect is achieved on astaxanthin, lutein, DHA and the like loaded with active nutritional ingredients, no additional artificial emulsifier is required, the potential safety hazard of the traditional superfine inorganic solid particles used as the Pickering emulsion emulsifier is overcome, the nutritional characteristics and stability of the microspheres are improved, and the purpose of synergistic interaction of nutrients is achieved.

Description

Active ingredient loaded nutrition microsphere and preparation method and application thereof
Technical Field
The invention relates to active ingredient loaded nutrient microspheres and a preparation method and application thereof, belonging to the field of food.
Background
The nutritive microspheres such as the popping beads are prepared by wrapping soluble fruit juice and other substances in the outer membrane by a special technology, and when a consumer eats the nutritive microspheres, teeth are slightly extruded, the outer membrane is broken, internal ingredients are popped out, and the product tastes peculiar and interesting and is impressive. The preparation method of the commonly used nutrient microsphere comprises the steps of dripping a sodium alginate material into a calcium ion-containing solution by using a drip forming machine for forming, cleaning and sterilizing. The nutritional microsphere popcom beads are highly favored by consumers due to the unique shape and bright color.
The fat-soluble component plays more and more roles in human health maintenance, and has wide application prospects in the aspects of health care products, medicines, cosmetics, food additives, aquaculture and the like. For example, astaxanthin is a fat-soluble ketocarotenoid widely existing in the biological world, and its ability to quench singlet oxygen and capture radicals is 10 or more times higher than that of β -carotene and 100 times stronger than that of vitamin E, which is also called super vitamin E. It has the functions of inhibiting tumor, strengthening immunity, preventing cardiac and cerebral vascular diseases, eyeground pathological changes, etc. DHA is a fat-soluble polyunsaturated fatty acid essential to the human body and is mainly supplemented by food. DHA is an important constituent of brain cell membrane, participates in the formation and development of brain cells, increases the content of DHA in food, and thus is beneficial to enhancing the learning and memory functions, the healthy development of brain and nerves, and the prevention and treatment of senile dementia. Lutein, an important antioxidant, is a member of the carotenoid family, also known as "phytoxanthin", is the most important nutrient in the human retina, also known as "ocular gold", contains high concentrations of lutein in the macula of the eye, is the only carotenoid in the retina and lens of the eye, an element that is not manufactured by the human body itself and must be supplemented by external intake. In the absence of this element, the eye is blind.
In recent years, the use of Pickering emulsions in the fields of food, pharmaceutical, cosmetic and the like has received great attention. The Pickering emulsion is obtained by using ultrafine solid particles as an oil-in-water or water-in-oil type emulsifier, and the ultrafine solid particles conventionally used as an emulsifier are clay, silica, metal hydroxide, graphite, carbon black, and the like. If the solid particles are more wettable by the oil phase, the emulsion is of the W/O (water-in-oil) type; conversely, if the solid particles are more wettable by the aqueous phase, the emulsion is of the O/W (oil in water) type. The stability of Pickering emulsions is related to the concentration of solid particles, particle size, wettability, etc. How to prepare Pickering emulsion by using edible superfine solid particles, such as animal and plant source food protein, as an emulsifier has important significance for researching and developing nutritional food loaded with active ingredients.
Currently, commercially available nutritional microsphere products are heavily loaded with water-soluble fruit juices and the like, such as: patent CN 110301643A discloses a preparation method of astaxanthin-calcium alginate microspheres, which is to dissolve astaxanthin in olive oil to prepare an oil phase; dissolving lecithin in water to obtain water phase; mixing the water phase and the oil phase, dispersing at a high speed, homogenizing, adding sodium alginate to obtain a mixed solution, and dripping the mixed solution into the calcium ion solution in a spraying manner to obtain the astaxanthin-calcium alginate microspheres; however, the diameter of the microsphere prepared by the method is only 0.5-5 microns, and the lecithin is a small molecular phospholipid unsaturated fatty acid emulsifier, so that under normal conditions, light, temperature, metal ions, unknown impurities and the like can be used as catalytic conditions for oxidation reaction of lecithin, so that the oxidation is accelerated. Natural lecithin contains a large amount of unsaturated fatty acids and many impurities, and thus is very easily oxidized. Besides being protected from light and low temperature, the most important point for avoiding the oxidation of the phospholipid is to isolate the oxygen environment. Under the aerobic condition, even if the lecithin is stored at the temperature of 20 ℃ below zero in a sealed manner, the natural lecithin is easy to oxidize and deteriorate, and the phenomena of deepened phospholipid color, aggravated smell and overproof peroxide value are shown. Therefore, the stability and the antioxidant capacity of the conventional nutrient microspheres prepared by using lecithin as an emulsifier are not ideal, and further improvement is needed. Patent CN113455647A discloses a preparation method of a high-stability double emulsion and its application in astaxanthin carrying, mainly using a solution containing gelatin and other gel substances as an internal water phase W1, using a glycerol ester containing polyglycerol-ricinoleate (PGPR) as an oil phase O, and obtaining a W1/O emulsion under the condition of shearing emulsification; adding a water phase W2 with sodium caseinate dissolved into the primarily obtained W1/O emulsion, performing high shear treatment to obtain a W1/O/W2 double emulsion, adding a gel solution into the outermost layer of the formed double emulsion, and performing shear treatment again to obtain a final high-stability double emulsion; polyglycerol-ricinoleate (PGPR) is an artificially synthesized emulsifier, and is obtained from two raw materials: castor oil and glycerol, castor oil is the most expensive vegetable oil, glycerol is a natural polyol and is an essential component for all fats, PGPR is non-toxic, but there is still a strict limit of 5.0(g/kg) in mayonnaise and salad dressing, etc., and there is no report on the preparation of nutritional microspheres using double emulsion to encapsulate astaxanthin. Patent CN113826878A discloses a preparation technology for preparing double emulsion by using euglena powder, which requires adjusting pH of euglena powder aqueous dispersion with sodium hydroxide solution, heating, centrifuging to remove precipitate to obtain alkali extract, adjusting pH to isoelectric point with hydrochloric acid solution, centrifuging at low temperature, and collecting precipitate; washing with deionized water, dispersing, adjusting to neutral with sodium hydroxide solution, and lyophilizing to obtain euglena protein solution; finally, mixing the euglena protein solution with edible oil, and shearing at high speed to obtain W/O/W type double emulsion; although the types of emulsifiers used for preparing the double emulsion are reduced and the safety of the emulsion is improved, the steps are complicated, the stability of the emulsion is relatively weak, and the emulsion is not suitable for being used as a high internal phase emulsion.
Therefore, how to design and prepare the Pickering emulsion by using natural edible superfine protein particles as an emulsifier and encapsulate easily-oxidized fat-soluble nutritional ingredients such as astaxanthin, DHA, lutein and the like inside to prepare the nutritional microspheres, the Pickering emulsion and the nutritional microspheres are combined to resist the interference of external environmental factors such as heat, oxygen and the like, so that the gastric acid damage is effectively avoided, the dual-protection effect is exerted, and the method has important research value in the aspect of improving the synergistic capability of active nutritional ingredients.
Disclosure of Invention
[ problem ] to
Active fat-soluble nutrients are difficult to wrap, easy to break and unstable in the conventionally prepared nutrient microspheres, and the active nutrient components are sensitive to physical and chemical environmental factors such as heat and are easy to oxidize, so that the application of the active nutrient components in the food industry is greatly limited.
[ solution ]
In order to solve the problems, the invention adopts natural edible protein aqueous solution as water phase, oil as oil phase and natural edible protein as superfine particle emulsifier at the same time to prepare Pickering emulsion; then, by introducing grape seed extract, green tea extract, polysaccharide, sodium alginate and calcium ions, the microspheres generate protein-protein and alginic acid-calcium double cross-linking, and the stable millimeter-grade nutritional microspheres are obtained. The Pickering emulsion and the nutritional microspheres are combined, so that the double protection effect is achieved on astaxanthin, lutein, DHA and the like loaded with active nutritional ingredients, no additional artificial emulsifier is needed, the potential safety hazard of the traditional superfine inorganic solid particles used as the Pickering emulsion emulsifier is overcome, the nutritional characteristics and stability of the microspheres are improved, the purpose of synergistic interaction of nutrients is achieved, and the application and popularization value is important in the aspect of improving the utilization degree of the active nutritional ingredients. Moreover, the nutritional microspheres are applied to cakes, quick-frozen rice and flour products, quick-frozen prepared foods, cooked aquatic products and can products, do not crack in the high-temperature cooking process, and have better stability.
The first purpose of the invention is to provide a method for preparing Pickering emulsion loaded with active ingredients, wherein the method comprises the steps of taking natural edible protein aqueous solution as a water phase and grease as an oil phase, and carrying out high-speed shearing to obtain the Pickering emulsion loaded with the active ingredients;
wherein the natural edible protein aqueous solution comprises one or more of astaxanthin-containing Haematococcus pluvialis protein aqueous solution, chia seed protein aqueous solution, Chlorella pyrenoidosa protein aqueous solution, bonito elastin aqueous solution, and egg white powder aqueous solution; the oil comprises one or more of DHA algae oil, fish oil, gamma-linolenic acid oil, krill oil, chia seed oil, lutein ester oil solution, and soybean oil.
In one embodiment of the invention, the concentration of the natural edible protein aqueous solution is 2-5 wt%.
In one embodiment of the present invention, the method for preparing the astaxanthin-containing haematococcus pluvialis protein aqueous solution comprises:
dispersing haematococcus pluvialis powder containing astaxanthin in a NaCl solution to obtain a dispersion liquid; ultrasonically breaking the wall of the dispersion liquid in water bath ultrasound, and centrifuging to remove precipitate to obtain supernatant; adding ammonium sulfate powder into the supernatant, and centrifuging at low temperature to collect precipitate after the ammonium sulfate powder is completely dissolved; dispersing the precipitate with PBS solution to obtain protein solution of haematococcus pluvialis powder containing astaxanthin, and evaporating to required concentration; wherein the ratio of the haematococcus pluvialis powder containing astaxanthin to the NaCl solution is 1: 10-1: 50 g/mL; the concentration of the NaCl solution is 1 to 5 weight percent; the ultrasonic temperature is 25-45 ℃, and the ultrasonic time is 1-2 h; centrifuging at 10000-12000 rpm for 4-6 min at 4 ℃; the evaporation is rotary evaporation, the temperature is 30-45 ℃, the vacuum degree is 0.07-0.1 MPa, and the time is 60-90 min.
In one embodiment of the present invention, the preparation method of the chlorella pyrenoidosa protein aqueous solution comprises the following steps:
dispersing the chlorella protein powder in a NaCl solution to obtain a dispersion liquid; ultrasonically breaking the wall of the dispersion liquid in water bath ultrasound, and centrifuging to remove precipitate to obtain supernatant; adding ammonium sulfate powder into the supernatant, and after complete dissolution, centrifuging at low temperature and collecting precipitate; dispersing the precipitate with PBS solution to obtain Chlorella pyrenoidosa protein solution, and evaporating to required concentration; wherein the ratio of the chlorella pyrenoidosa powder to the NaCl solution is 1: 10-1: 50 g/mL; the concentration of the NaCl solution is 1 to 5 weight percent; the ultrasonic temperature is 25-45 ℃, and the ultrasonic time is 1-2 h; centrifuging at 10000-12000 rpm for 4-6 min at 4 ℃; the evaporation is rotary evaporation, the temperature is 30-45 ℃, the vacuum degree is 0.07-0.1 MPa, and the time is 60-90 min.
In one embodiment of the present invention, the preparation method of the chia seed protein aqueous solution comprises:
dispersing chia seed powder in a NaCl solution to obtain a dispersion liquid; ultrasonically breaking the wall of the dispersion liquid in water bath ultrasound, and centrifuging to remove precipitate to obtain supernatant; adding ammonium sulfate powder into the supernatant, and after complete dissolution, centrifuging at low temperature and collecting precipitate; dispersing the precipitate with PBS solution to obtain chia seed protein solution, and evaporating to required concentration; wherein the ratio of chia seed powder to NaCl solution is 1: 10-1: 50 g/mL; the concentration of the NaCl solution is 1 to 5 weight percent; the ultrasonic temperature is 25-45 ℃, and the ultrasonic time is 1-2 h; centrifuging for 4-6 min at 10000-12000 rpm under the condition of 4 ℃; the evaporation is rotary evaporation, the temperature is 30-45 ℃, the vacuum degree is 0.07-0.1 MPa, and the time is 60-90 min.
In one embodiment of the present invention, the volume ratio of the water phase to the oil phase is 50% to 30% to 70%, more preferably 35% to 65% to 30% to 70%.
In one embodiment of the present invention, the high speed shearing is 10000-12000 rpm for 3-5 min.
The second purpose of the invention is to prepare the Pickering emulsion loaded with active ingredients by the method.
The third purpose of the invention is to provide a method for preparing edible nutritional microspheres based on the Pickering emulsion loaded with active ingredients, which comprises the following steps:
mixing the Pickering emulsion loaded with the active ingredients, polysaccharide and a sodium alginate solution, then adding a grape seed extract and a green tea extract, and uniformly mixing to obtain a mixture; and then dripping the mixture into a calcium ion solution to react to form protein-protein and alginic acid-calcium double cross-linking, thereby obtaining the edible nutritional microspheres.
In one embodiment of the invention, the polysaccharide comprises one or more of plantain seed husk powder, pseudo-microsphere algae powder, mussel polysaccharide, inulin, chitosan oligosaccharide and chia seed polysaccharide.
In one embodiment of the invention, the volume ratio of the Pickering emulsion loaded with the active ingredient to the sodium alginate solution is 1: 1.
In one embodiment of the invention, the addition amount of the polysaccharide is 0.5-5% of the total mass of the Pickering emulsion and the sodium alginate solution loaded with the active ingredients.
In one embodiment of the invention, the mass concentration of the sum of the grape seed extract and the green tea extract in a mixed solution of Pickering emulsion loaded with active ingredients, polysaccharide and sodium alginate solution is 2-3 wt%.
In one embodiment of the invention, the mass ratio of the grape seed extract to the green tea extract is 1: 1-3.
In an embodiment of the invention, the calcium ion solution is one of a calcium chloride solution, a calcium lactate solution, a calcium sulfate solution, and a calcium acetate solution, the mass concentration of the calcium ion solution is 1-4%, and the solvent of the calcium ion solution is water.
In one embodiment of the present invention, the dropping rate of the mixture is 0.5 to 10 mL/min.
In one embodiment of the invention, the reaction is carried out at 20-30 ℃ for 3-5 h.
The fourth purpose of the invention is the edible nutrient microsphere prepared by the method.
In one embodiment of the invention, the particle size of the edible nutritional microspheres is 3-8 mm.
The fifth purpose of the invention is to apply the Pickering emulsion and the edible nutritional microspheres loaded with the active ingredients in the field of food.
In one embodiment of the invention, the food comprises cakes, quick-frozen rice and flour products, quick-frozen prepared foods, cooked aquatic products, canned products and the like.
In one embodiment of the invention, the application is the addition of edible nutritional microspheres to a filling.
[ advantageous effects ]
(1) The Pickering emulsion containing the active nutrient components has good stability, is suitable for embedding fat-soluble nutrient components such as astaxanthin, DHA, lutein and the like, improves the bioavailability of functional factors, has higher safety and no toxic action on human bodies, can be applied to the food fields such as cakes, quick-frozen rice and flour products, quick-frozen prepared foods, cooked aquatic products, canned products and the like, and is not easily influenced by factors such as the pH value, the salt concentration, the temperature, the oil phase composition and the like of a system.
(2) Compared with the raw materials of the existing Pickering emulsion, the protein base material containing the active ingredients used by the invention is derived from animal and plant food raw materials, has the characteristics of safety, nutrition, health and the like, and protects the active nutritional ingredients contained in the emulsion. In addition, astaxanthin is the king of oxidation resistance in the nature; the algae oil, the fish oil, the gamma-linolenic acid grease, the krill oil and the chia seed oil are rich in DHA (docosahexaenoic acid), which is also called as brain gold and has the effects of improving memory, protecting eyesight and the like; lutein, also known as "eye gold", has an important protective effect on eyes.
(3) Conventionally, most of active substances such as DHA, astaxanthin, lutein and the like are fat-soluble active substances, the taste is poor when the active substances are mixed in stuffing, and the activity is rapidly reduced after heating for more than 5 minutes at 100 ℃, so that the inherent efficacy is lost. The edible nutritional microspheres of the invention are applied to food, so that the loss of nutritional ingredients in high-temperature cooking can be reduced, and the color is unchanged.
(4) Active substances such as lutein, astaxanthin and the like mostly have natural and bright colors, and the color of the stuffing can be changed by directly mixing the active substances with the stuffing, so that the sensory judgment and the preference of people are influenced. The nutritional microspheres loaded with active ingredients and the stuffing exist independently, so that the taste and color judgment of consumers are not influenced.
(5) The invention provides a preparation method of nutrient microspheres, which is simple and convenient to operate, short in time consumption, simple in required equipment and suitable for industrial large-scale production; the nutritional microspheres loaded with the active substances keep good integrity in the stomach and are not damaged by gastric acid, and the loaded active substances can be quickly released in the intestinal tract, so that the active substances are effectively released in the intestinal tract.
(6) Protein-stabilized Pickering emulsions containing active substances, which are doubly cross-linked by protein-protein and calcium alginate, are not reported. The Pickering emulsion has good stability and high safety, and can be used for stabilizing active nutrient components; the emulsion is prepared into the edible nutritional microspheres, is applied to the food fields of cakes, quick-frozen rice and flour products, quick-frozen prepared foods, cooked aquatic products, canned products and the like, has better stability in the heating and cooking process, and has larger application potential.
Drawings
FIG. 1 is an appearance diagram of Pickering emulsion obtained by using 50% to 50% volume ratio, 45% to 55% volume ratio, 40% to 60% volume ratio, 35% to 65% volume ratio and 30% to 70% volume ratio of water phase and oil phase.
FIG. 2 is a micrograph of Pickering emulsion obtained with the aqueous phase and the oil phase in a volume ratio of 50% to 50% (a), 45% to 55% (b), 40% to 60% (c), 35% to 65% (d), 30% to 70% (e).
FIG. 3 is a graph showing the appearance of Pickering emulsions obtained with a volume ratio of aqueous phase to oil phase of 50% to 50%, 45% to 55%, 40% to 60%, 35% to 65%, 30% to 70% on different days of storage.
FIG. 4 is an appearance diagram of Pickering emulsions of comparative example 1(a) and example 1 (b).
FIG. 5 is an appearance view (a), a size distribution view (b) and a micrograph (c) of the Pickering emulsion of example 1 under freezing conditions (-20 and 4 ℃) and heating conditions (37, 65 and 90 ℃).
FIG. 6 is a schematic representation of the edible nutritional microspheres of example 3 after being uncooked (a) and boiled at 100 ℃ for 10 minutes (b).
FIG. 7 is an appearance diagram of the Pickering emulsion obtained in example 3 and comparative example 6, wherein a is the appearance diagram of the Pickering emulsion obtained with the volume ratio of the water phase to the oil phase being 35% to 65%; b is an appearance diagram of the Pickering emulsion obtained by placing the Pickering emulsion for 2 hours, wherein the volume ratio of the water phase to the oil phase is 35 percent to 65 percent; c is a nutrition microsphere prepared by the emulsion obtained in the step a; d is the volume ratio of the oil phase to the sodium alginate of 1:1, fully mixing; e is the volume ratio of oil phase to sodium alginate of 1:1, fully mixing and standing for 1 minute; f is the phenomenon that the oil phase and the sodium alginate mixed solution which are fully mixed d are dripped into calcium chloride and are kept stand for 30 min.
FIG. 8 is an appearance view (a) of the edible nutritional microspheres of example 3 applied to the filling of dumplings without boiling and an appearance view (b) of the boiled microspheres after boiling at 100 ℃ for 10 minutes.
Detailed Description
The following description of the preferred embodiments of the present invention is provided for the purpose of better illustrating the invention and is not intended to limit the invention thereto.
The test method comprises the following steps:
and (3) testing the embedding rate: diluting 50 mu L of the emulsion in 4.95mL of mixed solution of dichloromethane and methanol (2:1, v/v), uniformly mixing by vortex, centrifuging for 20min at 4472g, and measuring the absorbance of the supernatant at 480nm by an ultraviolet spectrophotometer to obtain the astaxanthin content in the emulsion under different conditions.
Testing oil leakage rate: and centrifuging the emulsions under different conditions at 8000rpm for 2min, and calculating the ratio of the oil mass separated out from each emulsion after centrifugation to the total emulsion volume.
Testing of emulsion stability: the emulsions under different conditions were stored for 14 days, during which time it was observed whether or not a water layer separated out at the bottom of each emulsion, and no separation out indicates good stability.
Oil retention test: dispersing the protein powder into oil, fully stirring, centrifuging at 8000rpm for 2min, removing supernatant, collecting precipitate, weighing, and calculating weight gain percentage of the protein powder to obtain the holding rate.
Testing of temperature stability: the emulsions under different conditions were placed at-20, 4, 37, 65 and 90 ℃ for 14 days, during which time it was observed whether or not a water layer had separated out at the bottom of each emulsion, and a large amount of the emulsion was poorly exuded, a small amount of the emulsion was generally exuded, and no separation was observed, indicating that the stability was good.
In the examples, the terms "percent" and "percent" are not specifically defined; the solvent of the solution is not particularly limited, and water is used as the solvent.
Example 1
A method for preparing an active ingredient loaded Pickering emulsion comprising the steps of:
(1) water phase:
adding 20g of astaxanthin-containing haematococcus pluvialis powder into 400mL of 2 wt% NaCl solution, and stirring at room temperature until the haematococcus pluvialis powder is uniformly dispersed to obtain astaxanthin-containing haematococcus pluvialis dispersion liquid; ultrasonically breaking the walls of haematococcus pluvialis dispersion liquid containing astaxanthin in water bath ultrasonic at the temperature of 45 ℃ for 2 hours; centrifuging the dispersion obtained after ultrasonic treatment at 4 ℃ and 7000rpm for 15min, and collecting the supernatant; adding ammonium sulfate powder with saturation degree of 90% (i.e. 0.662g/mL) into the supernatant, stirring at room temperature, after completely dissolving, centrifuging at 4 deg.C and 12000rpm for 5min, and collecting precipitate; adding the precipitate into 50mL of PBS solution, and uniformly dispersing to obtain a haematococcus pluvialis protein solution containing astaxanthin; rotationally evaporating astaxanthin-containing haematococcus pluvialis protein solution at 45 ℃ under the vacuum degree of 0.1MPa to ensure that the astaxanthin-containing haematococcus pluvialis protein concentration is 2.0 wt% to be used as a water phase;
(2) oil phase:
taking 12mL of DHA algae oil as an oil phase;
(3) pickering emulsion:
and (3) mixing the water phase obtained in the step (1) and the oil phase obtained in the step (2) according to the volume ratio of 35% to 65%, and shearing at 10000rpm for 3min to obtain the Pickering emulsion loaded with active ingredients (astaxanthin and DHA).
Example 2
The volume ratio of the aqueous phase to the oil phase in example 1 was adjusted to 50%, 45% to 55%, 40% to 60%, 35% to 65% (example 1), 30% to 70%, and the rest was kept the same as in example 1, to obtain Pickering emulsion.
The obtained Pickering emulsion was subjected to a performance test, and the test results were as follows:
FIG. 1 is an appearance diagram of Pickering emulsions obtained with a volume ratio of aqueous phase to oil phase of 50% to 50%, 45% to 55%, 40% to 60%, 35% to 65%, 30% to 70%, as can be seen in FIG. 1: with the increase of the oil phase from 50% to 70%, the Pickering emulsion can be formed under the condition that each water phase is compared with the oil phase, and the emulsion has no layering phenomenon, which indicates that the prepared Pickering emulsion has a wider water phase-oil phase stable range, wherein the oil phase is more stable at 65% and 70%, and the oil-water layering phenomenon is not seen when the Pickering emulsion is placed at room temperature.
FIG. 2 is a micrograph of Pickering emulsions obtained with a volume ratio of aqueous phase to oil phase of 50% to 50%, 45% to 55%, 40% to 60%, 35% to 65%, 30% to 70%, as can be seen in FIG. 2: as the oil phase increases from 50% to 70%, the diameter of the oil-in-water droplets gradually decreases, the spacing between the droplets decreases, the structure becomes more compact, and 65% and 70% of the oil phase droplets are most densely structured.
FIG. 3 is a graph of the appearance of Pickering emulsions obtained with a volume ratio of aqueous phase to oil phase of 50% to 50%, 45% to 55%, 40% to 60%, 35% to 65%, 30% to 70% on different days of storage; as can be seen from fig. 3: the Pickering emulsion prepared under the conditions of the water phase-oil phase ratio is stable when placed for two days, and from 7 th day to 14 th day, the Pickering emulsion with the oil phase ratio of 50%, 55% and 60% has oil-water demixing phenomenon, which shows that the Pickering emulsion with the oil phase ratio of 65% and 70% is most stable when placed at room temperature for two weeks.
Comparative example 1
Spray drying the haematococcus pluvialis protein solution containing astaxanthin in the example 1 to obtain haematococcus pluvialis protein powder containing astaxanthin;
and (3) carrying out spray drying on the soybean protein isolate to obtain the soybean protein isolate powder.
The obtained protein powder was tested for oil holding capacity, and the test results are shown in table 1:
TABLE 1
Protein powder Haematococcus pluvialis protein powder containing astaxanthin Isolated soy protein powder
Oil retention property (g/g) 3.59 2.24
As can be seen from table 1: the oil holding rate of the haematococcus pluvialis protein powder containing astaxanthin is 3.59g/g, which is improved by 60.3% compared with 2.24g/g of soybean protein isolate, and the haematococcus pluvialis protein powder has better oil holding capacity.
Fig. 4 is an appearance diagram of Pickering emulsions of comparative example 1 and example 1. As can be seen from fig. 4: the emulsion of comparative example 1 had more foam and exhibited a thinning appearance on the surface, whereas the Pickering emulsion prepared using Haematococcus pluvialis protein containing astaxanthin was more viscous.
FIG. 5 is an appearance view (a), a size distribution view (b) and a micrograph (c) of the Pickering emulsion of example 1 under freezing conditions (-20 and 4 ℃) and heating conditions (37, 65 and 90 ℃); as can be seen from fig. 5: under the condition of variable temperature, the Pickering emulsion in the embodiment 1 is stable, the oil-water separation phenomenon does not occur, the emulsion is slightly influenced by the temperature, and the emulsion can still keep stable within the range of-20 to 90 ℃.
Comparative example 2
The DHA algal oil of example 1 was replaced with corn oil, and the rest was identical to example 1, to obtain Pickering emulsion.
Comparative example 3
The concentration of astaxanthin-containing Haematococcus pluvialis protein was adjusted to 1.5 wt%, and other conditions or parameters were the same as those in example 1.
The Pickering emulsions of example 1 and comparative examples 2 and 3 were subjected to performance testing, the results of which are as follows:
TABLE 2
Figure BDA0003707859690000091
Example 3
A method of preparing edible nutritional microspheres based on the Pickering emulsion of example 1, comprising the steps of:
mixing 5mL of Pickering emulsion of example 1, mussel polysaccharide and 5mL of 1% sodium alginate solution, adding grape seed extract and green tea extract (the mass ratio of the two is 1:2), wherein the sum of the grape seed extract and the green tea extract is 2.5 wt% in the mixed solution containing the Pickering emulsion, the mussel polysaccharide and the sodium alginate solution, and the adding amount of the mussel polysaccharide is 1.0 wt% of the total mass of the Pickering emulsion and the sodium alginate solution to obtain a mixture; and then dripping the mixture into a calcium lactate solution with the mass concentration of 2% at the speed of 0.5mL/min, reacting for 4 hours at room temperature to form protein-protein and alginic acid-calcium double cross-linked, filtering, and collecting the edible nutritional microspheres with the size of millimeter.
And (3) carrying out performance test on the obtained nutrition microspheres, wherein the test result is as follows:
FIG. 6 is a schematic representation of the edible nutritional microspheres of example 3 after being uncooked (a) and boiled at 100 ℃ for 10 minutes (b), as can be seen in FIG. 6: the shape and the size of the nutrient microsphere are uniform, the particle size is about 3-8 mm, the nutrient microsphere is complete after boiling for 10min, and the phenomenon of crushing is not seen.
Dispersing the edible nutritional microspheres, astaxanthin-containing Haematococcus pluvialis powder and Haematococcus pluvialis protein of example 3 in water, heating in a water bath at 95 ℃ for 10 minutes, measuring the astaxanthin content before and after heating, and calculating the astaxanthin loss rate after heating;
and (3) heating the DHA algae oil and the edible nutritional microspheres in the example 3 in a water bath at 95 ℃ for 10 minutes, measuring the DHA content before and after heating, and calculating the DHA loss rate after heating.
The results are shown in tables 3 to 4:
as can be seen from table 3: example 3 the heating loss rate of astaxanthin after being made into nutritional microspheres by Pickering emulsion was significantly lower than that of Haematococcus pluvialis powder and Haematococcus pluvialis protein alone.
As can be seen from table 4: after the nutrient microspheres are prepared from the Pickering emulsion, the heating loss rate of DHA in the nutrient microspheres is obviously lower than that of single unprotected DHA algal oil, which shows that the nutrient microspheres prepared in example 3 have an important protection effect on stabilizing active nutrient components such as astaxanthin and DHA.
TABLE 3 conditions of astaxanthin before and after heating under the protection of Pickering emulsion microspheres
Figure BDA0003707859690000101
TABLE 4 DHA before and after Pickering emulsion microsphere protection heating
Figure BDA0003707859690000102
Comparative example 4
A method for preparing edible nutritional microspheres through single cross-linking, which comprises the following steps:
mixing 5mL of Pickering emulsion of example 1, mussel meat polysaccharide and 5mL of 1% sodium alginate solution, wherein the addition amount of the mussel meat polysaccharide is 1.0 wt% of the total mass of the Pickering emulsion and the sodium alginate solution, so as to obtain a mixture; and then dripping the mixture into a calcium lactate solution with the mass concentration of 2% at the speed of 0.5mL/min, reacting for 4 hours at room temperature to form alginic acid-calcium cross-linked, filtering, and collecting the edible nutritional microspheres with the sizes of millimeter grades.
Comparative example 5
The haematococcus pluvialis protein concentration containing astaxanthin in example 1 is 1.0 wt%, and the rest is consistent with that in example 1, so that Pickering emulsion is obtained; edible nutritional microspheres were then prepared according to example 3.
The edible nutritional microspheres prepared in example 3 and comparative examples 4 and 5 were tested and the results are shown in table 5 below:
TABLE 5
Figure BDA0003707859690000103
Comparative example 6
A method of preparing edible nutritional microspheres comprising the steps of:
12mL of DHA algal oil and sodium alginate solution are mixed in a volume ratio of 1:1, fully mixing to obtain a mixed solution; and then dripping the mixed solution into a calcium lactate solution with the mass concentration of 2% at the speed of 0.5mL/min, reacting for 4 hours at room temperature, filtering, and collecting the edible nutritional microspheres with the sizes of millimeter grades.
The microspheres obtained in example 3 and comparative example 6 were subjected to performance tests, and the test results were as follows:
in fig. 7, a represents that the volume ratio of the water phase to the oil phase is 35%: appearance of 65% of the obtained Pickering emulsion; b is the volume ratio of the water phase to the oil phase is 35%: appearance diagram of 65% of the obtained Pickering emulsion after being placed for 2 hours; c is the nutritional microspheres prepared using the emulsion obtained in a (example 3) without exudation of the oil phase; d is the volume ratio of the oil phase to the sodium alginate solution of 1:1, fully mixing; e is the volume ratio of the oil phase to the sodium alginate solution of 1:1, fully mixing and standing for 1 minute, then generating a layering phenomenon, and separating oil from water; f is that the oil phase and the sodium alginate mixed solution which are fully mixed d are dripped into calcium chloride, and the oil in the microspheres (comparative example 6) is separated out after standing for 30 min. The conventionally prepared nutrition microspheres are difficult to wrap active fat-soluble nutrients directly, are easy to break and are unstable, and the lipid-soluble active nutrition ingredients can be stably encapsulated in the microspheres by introducing Pickering emulsion, so that the stability is improved.
Example 4
Adding the edible nutritional microspheres of example 3 into dumpling stuffing, making into dumplings, and decocting with boiling water for 10 min.
The appearance of the boiled dumplings is shown in figure 8, and can be seen from figure 8: after boiling, the edible nutritional microspheres can still keep the original shape and color and have no cracking phenomenon, which shows that the prepared edible nutritional microspheres of the Pickering emulsion containing astaxanthin and DHA are not easily affected by high temperature, have good heat resistance and higher stability.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

Claims (10)

1. A method for preparing Pickering emulsion loaded with active ingredients is characterized in that natural edible protein aqueous solution is used as a water phase, grease is used as an oil phase, and the Pickering emulsion loaded with the active ingredients is obtained by high-speed shearing;
wherein the natural edible protein aqueous solution comprises one or more of astaxanthin-containing Haematococcus pluvialis protein aqueous solution, chia seed protein aqueous solution, Chlorella pyrenoidosa protein aqueous solution, bonito elastin aqueous solution, and egg white powder aqueous solution; the oil comprises one or more of DHA algae oil, fish oil, gamma-linolenic acid oil, krill oil, chia seed oil, lutein ester oil solution, and soybean oil.
2. The method according to claim 1, wherein the concentration of the aqueous solution of the natural edible protein is 2 to 5 wt%.
3. The method of claim 1 wherein the volume ratio of the aqueous phase to the oil phase is 50% to 30% to 70%.
4. The method of claim 1, wherein the high shear is 10000-12000 rpm for 3-5 min.
5. The Pickering emulsion loaded with the active ingredients and prepared by the method of any one of claims 1 to 4.
6. A method for preparing edible nutritional microspheres based on the Pickering emulsion loaded with active ingredients of claim 5, characterized by comprising the following steps:
mixing the Pickering emulsion loaded with the active ingredients, polysaccharide and a sodium alginate solution, then adding the grape seed extract and the green tea extract, and uniformly mixing to obtain a mixture; and then dripping the mixture into a calcium ion solution to react to form protein-protein and alginic acid-calcium double cross-linking, thereby obtaining the edible nutritional microspheres.
7. The method of claim 6, wherein the polysaccharide comprises one or more of psyllium husk powder, microalgal powder, mussel polysaccharide, inulin, chitooligosaccharide, chia seed polysaccharide; the mass ratio of the grape seed extract to the green tea extract is 1: 1-3.
8. Edible nutritional microspheres produced by the process of claim 6 or 7.
9. The edible nutritional microspheres of claim 8, wherein the particle size of the edible nutritional microspheres is 3-8 mm.
10. The Pickering emulsion loaded with active ingredients as claimed in claim 5 and the edible nutritional microspheres as claimed in claim 8 are applied to the field of food.
CN202210713518.4A 2022-06-22 2022-06-22 Active ingredient loaded nutrition microsphere and preparation method and application thereof Pending CN115024493A (en)

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