Disclosure of Invention
The invention aims to provide a duck fatty liver microcapsule and a preparation method thereof, thereby making up the defects of the prior art.
The applicant is dedicated to research on a suitable method for preparing the duck fatty liver whole liver powder microcapsules, and through long-time research and screening of four factors of the emulsifier adding amount, the core-wall material proportion, the reaction pH value and the embedding temperature in the microcapsule preparation process, the optimal process condition for preparing the duck fatty liver microcapsules under the assistance of ultrasonic waves is determined.
The preparation method of the duck fatty liver powder microcapsule comprises the steps of emulsion preparation and vacuum freeze drying:
1) preparation of an emulsion: fully mixing and dissolving core material duck fat liver whole liver serous fluid in a wall material solution to prepare an emulsion, and adjusting the pH value and the temperature of the emulsion to emulsify;
wherein the wall material is gelatin and Arabic gum aqueous solution, and the concentration of the wall material solution is 12.5%; preferably, the mass ratio of the gelatin to the Arabic gum is 1: 1;
the mass ratio of the core material to the wall material in the emulsion is 1: 1-5; preferably 1: 1.29;
the pH value of the emulsion preparation is 1.0-5.0, and the optimal pH value is 4.34; the preparation temperature is 43.71 ℃;
preferably, alkyl glucoside (APG) with the weight of 1.25 percent of the total weight of the emulsion is added to serve as an emulsifier when the emulsion is prepared;
furthermore, chitosan accounting for 1 percent of the total mass of the emulsion is added in the preparation of the emulsion;
2) vacuum freeze drying treatment: and precooling the emulsion, and then putting the emulsion into a freeze dryer for vacuum freeze drying to finish the preparation of the duck fatty liver powder microcapsules.
The vacuum freeze drying conditions are-40 deg.C, 15 hr and 100 Pa.
Compared with the prior art, the method has the following advantages:
1. the combination of the coating wall materials has synergy. Two natural or synthetic polymer polyelectrolytes with opposite charges are used as wall materials, core materials are dispersed in a wall material solution, the electrolyte generates electrostatic action by changing the solution condition, the solubility of the wall materials is reduced, and the wall materials are coagulated from the solution to coat the core materials to form the microcapsule.
2. The risk of active material loss is circumvented. The freeze drying method adopted by the invention can avoid the damage of high temperature to unsaturated fatty acid, effectively prevent the loss of unsaturated fatty acid in the preparation process, and improve the stability and convenience to a certain extent.
3. The selected wall material is green and healthy. The acacia gum embedding agent used in the invention is natural gum prepared from plant juice extracted from acacia senegal and acacia seyal, is a complex glycoprotein and polysaccharide mixture, can be dissolved into a thick and tasteless viscous agent in an aqueous solution, has good emulsification property and film-forming property, and can effectively ensure the product quality. As the Arabic gum does not generate heat basically, the Arabic gum is used as a good water-soluble dietary fiber, has the function of reducing cholesterol in blood and improves the function of a duck fat liver microcapsule product.
The microcapsule coating is carried out on the duck fatty liver powder, so that the shelf life of the product is greatly prolonged, the blank of the product field is filled, and the microcapsule coating has important economic and social values for widening the duck fatty liver utilization market and guaranteeing the human health.
The specific implementation mode is as follows:
according to the invention, the duck fatty liver microcapsule is prepared by adopting ultrasonic assistance, gelatin and Arabic gum are selected as wall materials, duck fatty liver whole liver powder is selected as a core material, alkyl glucoside (APG) is used as an emulsifier, chitosan is used as a coating agent, a response surface test is designed by taking the oil embedding rate as an index and the addition amount of the emulsifier, the core-wall material ratio, the reaction pH and the embedding temperature as factors, so that the optimal preparation process condition for preparing the duck fatty liver microcapsule is obtained, and the embedding rate and the thermal stability of the prepared duck fatty liver microcapsule are detected.
The preparation method of the duck fatty liver microcapsule comprises the following steps:
1) preparing core material duck fat liver serous fluid: homogenizing duck fat liver to obtain slurry. The slurry has complex components, contains different proportions of hepatocyte tissues, blood vessels, capillary bile ducts, crusted hoof tissues and the like besides grease, and is difficult to ensure the consistency of the shape and the size of particles after pulping; the encapsulation of microcapsules is difficult to perform due to the differences in particle size, shape and affinity of the substance. Different from goose fat liver oil researched previously, the goose fat liver oil is easy to coat and consistent in particle size as long as the proportion of the core material to the wall material is selected because the goose fat liver oil is single in component.
2) Selecting wall materials: gum arabic is negatively charged; gelatin is an amphoteric polymer polyelectrolyte, has positive charges under acidic conditions and negative charges under weakly acidic and alkaline conditions, can generate complex coacervation reaction when being combined under certain pH conditions, and has good film-forming property and stretchability.
3) Treating a core material wall material: the core material and the wall material are respectively treated by 300W ultrasonic wave for 10 min. The embedding rate of the microcapsules prepared after treatment is obviously higher than that of an untreated sample, and the wall material is more easily dissolved in water after ultrasonic treatment, so that the concentration of the emulsion is greatly increased, the freeze-drying time is shortened, and the embedding rate and the production efficiency of the microcapsules are improved.
4) Preparation of an emulsion: the alkyl glucoside (APG) is a non-ionic surfactant which does not generate ions in aqueous solution, is easy to dissolve in water due to a functional group with strong affinity to water, and has good protective colloid and emulsification effect for preparing oil-in-water emulsion. Compared with the anionic surfactant, the water-soluble polymer has higher emulsifying capacity and certain hard water resistance.
The chitosan is a product of chitosan deacetylation, and macromolecules of the chitosan have active hydroxyl and amino, and the active hydroxyl and the amino have strong chemical reaction capacity. When the core material microcapsule is coated, the coating can be adhered to the surface of the core material, so that the tensile strength, the water resistance and the bursting strength can be enhanced, and the coating is not easy to break during processing.
5) And (3) freeze drying treatment: the sample is pre-cooled at-40 ℃, because according to the phase equilibrium theory in thermodynamics, the three-phase electricity (the coexistence of vapor, liquid and solid) temperature of water is 0.0098 ℃, the triple-phase point pressure is 609.3Pa (4.57mmHg), and in the phase change process of water, when the pressure is lower than the triple-phase point pressure, the solid ice can be directly converted into gaseous water vapor, namely ice crystals to sublimate.
Freeze drying means that a substance containing a large amount of water is frozen in advance to crystallize free water in the substance and freeze into a solid state, then ice crystals in the substance are sublimated under a high vacuum condition, and after the ice crystals are sublimated, part of adsorbed water in the substance is removed, and finally, a dry substance with the residual water content of about 1-4% is obtained. The material must be completely frozen before freeze-drying, and if the material is not completely frozen, a large amount of foam is generated in the drying process, which seriously hinders the sublimation of water and reduces the recovery rate of the material.
The method of the present invention will be described in detail with reference to the following embodiments.
Example 1: screening test for duck fat liver microencapsulation process conditions
(1) Preparation of emulsions
Respectively preparing arabic gum and gelatin into aqueous solutions with certain concentration, fully dissolving and mixing, adding a certain amount of alkyl glycoside (APG) and chitosan, and stirring to be uniform, wherein the solution is used as a reaction solution; slowly dripping the duck fat liver serous fluid into the reaction liquid, adjusting the pH value with acetic acid, and performing ultrasonic treatment to form an emulsion.
(2) Freeze drying treatment of emulsion
Pouring the emulsion into a plastic culture dish, precooling at-40 ℃, putting into a freeze dryer, freeze-drying for 15h, and taking out.
(3) Determination of encapsulation efficiency of microcapsules
The embedding rate refers to the ratio of the content of the embedded duck fatty liver serous fluid in the microcapsule product to the total amount of the duck fatty liver serous fluid added during embedding. The higher the embedding rate is, the larger the amount of the core material embedded is, and the better the effect is.
In the formula: w1: the total mass of the microcapsule duck fatty liver serous fluid; w2: the quality of the duck fat liver serous fluid on the surface of the microcapsule.
(4) Determination of optimum process for duck fat liver microcapsules
1) Screening of core material to wall material ratio
The core material and wall material ratios were set to 1:1, 1:2, 1:3, 1:4, and 1:5, respectively, at a reaction pH of 3.0 and a reaction temperature of 30 ℃ for testing, and the microcapsule embedding rate was measured. The proportion of the core material and the wall material is primarily screened.
2) Screening of reaction pH
The core material and wall material ratio was 1:1, and the reaction temperature was 30 ℃, and the reaction pH values were set to 1.0, 2.0, 3.0, 4.0, and 5.0, respectively, to perform the test, and the microcapsule embedding rate was measured. To carry out primary screening on the reaction pH value.
3) Screening of reaction temperature
The core material and wall material ratio was 1:1, and the reaction pH was 3.0, and the microcapsule embedding rate was measured by setting the reaction temperature to 10 ℃, 20 ℃, 30 ℃, 40 ℃ and 50 ℃. To perform a preliminary screening of the reaction temperature thereof.
4) Response surface test protocol and results
Table 1: response surface test factor coding and level
Table 2: response surface scheme and results
(5) Establishment and analysis of response surface regression model
The response surface software Design-Expert 8.0.6Trial analyzes the test result to obtain a linear regression equation as follows:
Y=57.61-1.77A+2.18B+3.85C-2.35AB+1.42AC+2.75BC-9.47A2-5.05B2-6.42C2
in the formula: a represents the core material-wall material ratio; b represents the reaction pH value; c represents the embedding temperature; y represents the embedding rate of the duck fat liver microcapsules.
Analysis of variance was performed on the above response surface test and the results are shown in table 1.3.
Table 3: response surface analysis of variance
As can be seen from Table 3, the value of Pr & gtF, and P & lt 0.05 of the ANOVA model are regarded as significant models, the fitting accuracy is good, and the response surface approximation model can be used for subsequent optimization design; the mismatching term Pr is more than the F value is more than 0.05, which shows that the mismatching is not obvious, the residual errors are all caused by random errors, the model is properly specified, and a regression equation can be used for replacing a test true point to analyze the test result, namely the model is well fitted in the whole regression region to be researched. Therefore, the regression equation can better describe the relationship between each factor and the response value, and the influence of each specific test on the response surface value is not a simple linear relationship.
The influence of the first item on the test is ranked as C > B > A, namely embedding temperature > reaction pH value > core wall material ratio, wherein the embedding temperature, the core material wall material ratio and the reaction pH value have significant influence on the test (P < 0.05). The significance of AB and BC in the interaction item is better, which shows that the interaction of the core material-wall ratio, the reaction pH value and the reaction temperature has larger influence on the embedding rate of the duck fat liver microcapsule.
After being analyzed by Design-Expert software, the ratio of the core material to the wall material is 1:1.29, the reaction pH value is 4.34, the reaction temperature is 43.71 ℃, and under the condition, the embedding rate of the duck fatty liver microcapsule is 58.79%.
Example 2: comparative experiment 1 for preparation process of duck fat liver microcapsules
The emulsion was prepared according to the procedure of example 1, maltodextrin and soy protein isolate were selected as coating wall materials, wherein the ratio of core material to wall material was 1:5, the reaction pH was 5.0, the wall reaction temperature was 60 ℃, the emulsifier alkyl glucoside was added, and the emulsion was formed by ultrasonic treatment. The emulsion is subjected to freeze drying treatment, the embedding rate of the microcapsules is measured according to the method, and after 5 times of repeated tests, the embedding rates of the obtained microcapsule products are 56.21%, 57.34%, 56.31%, 56.43% and 57.91%, which are lower than the embedding rate of 58.79% in example 1, and the effect of the maltodextrin and the isolated soybean protein as wall materials on coating the duck fatty liver powder is poorer than the effect of gelatin and Arabic gum as wall materials.
Example 3: comparative experiment 2 for preparation process of duck fat liver microcapsules
The preparation of the emulsion was carried out according to the procedure of example 1, selecting gelatin and gum arabic as coating wall material, and carrying out chitosan addition (1%) and non-addition treatments. Wherein the proportion of the core material to the wall material is 1:5, the reaction pH value is 5.0, the wall reaction temperature is 60 ℃, an emulsifier alkyl glucoside is added, and the mixture is subjected to ultrasonic treatment to form an emulsion. The emulsion was freeze-dried and the encapsulation efficiency of the microcapsules was determined as described above, each treatment being tested 5 times in duplicate, with the encapsulation efficiencies of the microcapsule products obtained without the addition of chitosan being 57.83%, 57.82%, 57.89%, 57.43% and 58.91%. The embedding rate of the microcapsule products obtained by adding chitosan is 63.77%, 63.81%, 63.80%, 63.44% and 64.94%; the coating rate is obviously higher than that of a group without chitosan, and is also obviously higher than that of 58.79% in example 1, which indicates that the chitosan added in the wall material can improve the embedding rate.
Example 4: comparative test for heat stability of duck fat liver microcapsules
Accurately weighing 2g of the microcapsule product prepared in the example 1 and the duck fat liver slurry, placing the microcapsule product and the duck fat liver slurry in a 105 ℃ oven for heating for 1, 2, 3, 4 and 5 hours respectively, weighing the mass reduction amount of each sample, and comparing the mass reduction amounts. The results show that the reduction of the microcapsule product and the duck liver serous fluid is increased in the whole process, but the reduction of the duck liver serous fluid is always higher than that of the microcapsule product, the reduction of the microcapsule product is relatively small after 4h, and the reduction of the duck liver serous fluid is always increased. This indicates that the heat stability of the microcapsule product is higher than that of duck fat liver serous fluid, i.e. the product has good heat stability.
The heat stability test of the microcapsule product shows that the stability of the microencapsulated duck fatty liver is superior to that of the duck fatty liver without microencapsulation. Thus, the microcapsule products prepared according to the process of the present invention have good thermal stability.
The invention adopts the coacervation phase separation method, the emulsion polymerization method and the freeze drying process to prepare the duck fatty liver microcapsule, effectively prevents the loss of unsaturated fatty acid in the preparation process, improves the stability to a certain extent and greatly prolongs the shelf life of the product. The core material and the wall material are respectively treated by ultrasonic waves, so that the wall material is more easily dissolved in water, and the freeze drying time is reduced. The selected wall material has good emulsification property and film forming property, and the embedding rate of the microcapsule is improved. Fills the blank in the research field of the duck fat liver embedding technology, fills the blank in the duck fat liver full liver powder microcapsule coating technology, and has important social and economic values for widening the utilization market space of the duck fat liver.