A kind of high biological activity sheep colostrum composite freeze-dried powder and preparation method thereof

Abstract

The invention discloses a kind of high biological activity sheep colostrum composite freeze-dried powder in the field of food technology, including the following components in weight: 60-70 parts of sheep colostrum powder, 5-8 parts of composite probiotic liposome freeze-dried powder, 6-9 parts of oligomannosaccharides, and the composite probiotic liposome freeze-dried powder is prepared by the following components: gallic acid, phytosterol, soybean phosphatidylcholine, modified chitosan, probiotic powder; probiotic powder is bifidobacterium longum powder or lactobacillus acidophilus powder. The present invention proposes to prepare modified phytosterol by esterification reaction of gallic acid and phytosterol, and prepare probiotic liposome by encapsulating probiotics with modified phytosterol and soybean phosphatidylcholine, and introduce carboxymethyl into chitosan to prepare modified chitosan, and prepare composite probiotic liposome freeze-dried powder with modified chitosan coated probiotic liposome, so as to protect probiotics, reduce the inactivation of probiotics, and improve its survival rate and colonization quantity in the intestine.

Description

Technical Field

[0001] The invention belongs to the field of food technology, and specifically refers to a high-bioactive sheep colostrum composite freeze-dried powder and a preparation method thereof. Background Art

[0002] Probiotics, as a group of active microorganisms in the human intestine that are beneficial to health, cover a variety of types, including lactic acid bacteria such as bifidobacterium, lactobacillus, streptococcus, enterococcus, and some non-lactic acid bacteria such as bacillus, yeast, propionibacterium, etc. These microorganisms play a vital role in the human body. They not only promote the digestion and absorption of nutrients, but also significantly enhance the body's immunity and help maintain the balance of intestinal flora. In addition, probiotics also have important functions such as improving antioxidant capacity, inhibiting intestinal inflammation, lowering serum cholesterol levels, and protecting intestinal mucosal barriers. However, the probiotic products currently on the market will be inactivated due to environmental changes during processing, transportation, storage, digestion and absorption. The number of probiotics that can truly colonize in the intestine in a living state is not enough to play its probiotic role. Therefore, how to ensure that probiotics are colonized in the human intestine with high activity and sufficient numbers is a problem that needs to be solved at present.

[0003] The existing technologies currently have the following main problems: it is difficult for probiotics to maintain high activity and sufficient quantity to enter the human intestine. Summary of the invention

[0004] In view of the above situation, in order to overcome the defects of the prior art, the present invention provides a high-bioactive sheep colostrum composite freeze-dried powder and a preparation method thereof. In order to solve the problem that probiotics are difficult to maintain high activity and sufficient quantity to enter the human intestine, the present invention proposes to prepare modified plant sterols by esterification reaction of gallic acid and plant sterols, and to prepare probiotic liposomes by encapsulating probiotics with modified plant sterols and soybean phosphatidylcholine. At the same time, carboxymethyl is introduced into chitosan to prepare modified chitosan, and probiotic liposomes are coated with modified chitosan to prepare composite probiotic liposome freeze-dried powder, thereby protecting probiotics, reducing the inactivation of probiotics, and improving their survival rate and colonization quantity in the intestine.

[0005] In order to achieve the above-mentioned purpose, the technical scheme adopted by the present invention is as follows: the present invention proposes a high-bioactive sheep colostrum composite freeze-dried powder, which comprises the following components in parts by weight: 60-70 parts of sheep colostrum powder, 5-8 parts of composite probiotic liposome freeze-dried powder, and 6-9 parts of oligomannosaccharides; the composite probiotic liposome freeze-dried powder is prepared from the following components: gallic acid, phytosterols, soybean phosphatidylcholine, modified chitosan, and probiotic powder; the probiotic powder is Bifidobacterium longum powder or Lactobacillus acidophilus powder.

[0006] Furthermore, the preparation method of the modified chitosan comprises the following steps:

[0007] S1, weigh chloroacetic acid and add it to isopropanol, the amount of chloroacetic acid added to isopropanol is 100 mg / mL, and stir until completely dissolved to obtain a chloroacetic acid-isopropanol solution;

[0008] S2. Weigh chitosan and add it to isopropanol, stir for 1-2 hours to obtain a mixed solution, then add sodium hydroxide solution, stir in a constant temperature water bath, slowly add the chloroacetic acid-isopropanol solution prepared in step S1, react for 2-3 hours, adjust the pH value to 7, wash, and dry to obtain modified chitosan.

[0009] Preferably, in step S2, the amount of chitosan added to the isopropanol is 100-150 mg / mL, the volume ratio of the mixed solution to the sodium hydroxide solution is 2:3, and the volume ratio of the mixed solution to the chloroacetic acid-isopropanol solution is 1:1-1.5.

[0010] Preferably, in step S2, the sodium hydroxide solution is a solution of sodium hydroxide in water, the mass fraction of the sodium hydroxide solution is 30%, the stirring temperature of the constant temperature water bath is 50-60° C., and the stirring speed is 100-200 rpm.

[0011] Furthermore, the preparation method of the composite probiotic liposome freeze-dried powder comprises the following steps:

[0012] (1) Weighing a certain amount of gallic acid and phytosterol and dissolving them in a reaction solvent, adding a catalyst, placing the mixture in a thermostat at 50° C. for 10-20 hours, separating and purifying the obtained product to obtain modified phytosterol;

[0013] (2) Weighing a certain amount of soybean phosphatidylcholine and the modified plant sterol prepared in step (1) and dissolving them in a chloroform-methanol mixture with a volume ratio of 1:1, placing them in a dark place at 25° C. for 15-30 minutes, removing the chloroform-methanol mixture by rotary evaporation, adding a certain amount of phosphate buffered saline solution and probiotic powder, and ultrasonically treating them in a constant temperature water bath to obtain probiotic liposomes;

[0014] (3) Weigh a certain amount of modified chitosan and dissolve it in water, add a certain amount of the probiotic liposome prepared in step (2), stir in a constant temperature water bath, perform ultrasonic treatment, and freeze-dry to obtain a composite probiotic liposome freeze-dried powder.

[0015] Preferably, in step (1), the molar ratio of the gallic acid to the phytosterol is 1:1.

[0016] Preferably, in step (1), the catalyst is p-toluenesulfonic acid, the molar ratio of the catalyst to phytosterol is 1:0.1, and the reaction solvent is toluene or tetrahydrofuran.

[0017] Preferably, in step (2), the weight ratio of the probiotic powder, the soybean phosphatidylcholine and the modified plant sterol is 1:8-10:2-4, the temperature of the constant temperature water bath is 40-50°C, the time of the ultrasonic treatment is 30-40 min, the power is 200 W, the volume ratio of the phosphate buffered saline solution to the chloroform-methanol mixture is 5-6:3, and the pH value of the phosphate buffered saline solution is 7.

[0018] Preferably, in step (3), the amount of the modified chitosan added to water is 100-150 mg / mL, the mass ratio of the modified chitosan to the probiotic liposome is 10:1, the stirring temperature of the constant temperature water bath is 40-50°C, the stirring speed is 100-200 rpm, the ultrasonic treatment time is 10-20 min, and the power is 200 W.

[0019] The invention provides a method for preparing a high-bioactivity goat colostrum composite freeze-dried powder, which specifically comprises the following steps: adding composite probiotic liposome freeze-dried powder and oligomannosaccharide into goat colostrum powder, stirring evenly, and obtaining the high-bioactivity goat colostrum composite freeze-dried powder.

[0020] The beneficial effects achieved by the present invention are as follows:

[0021] The present invention prepares modified phytosterols through esterification reaction of gallic acid and phytosterols, and prepares probiotic liposomes by encapsulating probiotics with modified phytosterols and soybean phosphatidylcholine, and introduces carboxymethyl into chitosan to prepare modified chitosan, and uses modified chitosan to encapsulate probiotic liposomes to prepare composite probiotic liposome freeze-dried powder, thereby protecting probiotics, reducing inactivation of probiotics, and improving their survival rate and colonization number in the intestine, so as to truly exert their probiotic effects; the wall material of conventional liposomes is mainly composed of phospholipids and cholesterol, but cholesterol is not suitable for people with cardiovascular diseases, phytosterols have a chemical structure similar to cholesterol, and can replace cholesterol to improve the stability and encapsulation rate of liposomes, and phytosterols are esterified with gallic acid. Modification can enhance the antioxidant effect of phytosterols and prevent the composite probiotic liposome freeze-dried powder from oxidative deterioration during processing, storage and transportation, thereby improving the storage stability of the composite probiotic liposome freeze-dried powder; introducing carboxymethyl groups into the molecular structure of chitosan to prepare modified chitosan can enhance the water solubility of chitosan, thereby facilitating the subsequent preparation of composite probiotic liposome freeze-dried powder. Since liposomes have a potential risk of damage during the freeze-drying process, the composite probiotic liposome freeze-dried powder is prepared by coating probiotic liposomes with modified chitosan, which can protect the probiotic liposomes during the freeze-drying process, thereby reducing the effect of freeze-drying on the probiotic liposomes; oligomannose can promote the proliferation of intestinal probiotic flora in the body and has the effect of improving human intestinal function. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The figure is a test result of the encapsulation efficiency of the composite probiotic liposome freeze-dried powder prepared in Examples 1-3 of the present invention and Comparative Examples 1 and 2;

[0023] Figure 2 The graph is a test result of storage stability of the composite probiotic liposome freeze-dried powder prepared in Examples 1-3 of the present invention and Comparative Examples 1 and 2;

[0024] Figure 3 The results of simulated gastric fluid test of the composite probiotic liposome freeze-dried powder prepared in Examples 1-3 of the present invention and Comparative Examples 1 and 2 are shown;

[0025] Figure 4 This is a diagram showing the results of a simulated digestion test of the composite probiotic liposome freeze-dried powder prepared in Examples 1-3 of the present invention;

[0026] Figure 5 The result of the redispersibility test of the composite probiotic liposome freeze-dried powder prepared in Examples 1-3 of the present invention is shown in FIG.

[0027] Figure 6 This is a transmission electron micrograph of the composite probiotic liposome freeze-dried powder prepared in Example 1 of the present invention.

[0028] The accompanying drawings are used to provide further understanding of the present invention and constitute a part of the specification. They are used to explain the present invention together with the embodiments of the present invention and do not constitute a limitation of the present invention. DETAILED DESCRIPTION

[0029] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments; based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0030] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein may be applied to the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only and are not intended to limit the content of this application.

[0031] The experimental methods in the following examples are conventional methods unless otherwise specified; the experimental materials and test strains used in the following examples are purchased from commercial channels unless otherwise specified.

[0032] Example 1

[0033] A high-bioactive sheep colostrum composite freeze-dried powder comprises the following components in parts by weight: 60 parts of sheep colostrum powder, 5 parts of composite probiotic liposome freeze-dried powder, and 6 parts of oligomannosaccharides.

[0034] The preparation method of modified chitosan comprises the following steps:

[0035] S1, weigh 3g of chloroacetic acid, add it to 30mL of isopropanol, and stir until it is completely dissolved to obtain a chloroacetic acid-isopropanol solution;

[0036] S2. Weigh 2 g of chitosan and add it to 20 mL of isopropanol. Stir for 1 h to obtain a mixed solution. Then add 30 mL of a 30% sodium hydroxide solution by mass, stir in a constant temperature water bath at 50°C at a speed of 100 rpm, slowly add 20 mL of the chloroacetic acid-isopropanol solution prepared in step S1, react for 2 h, adjust the pH value to 7, wash, and dry to obtain modified chitosan.

[0037] The preparation method of the composite probiotic liposome freeze-dried powder comprises the following steps:

[0038] (1) Weigh 10 mmol of gallic acid and 10 mmol of phytosterol and dissolve them in 50 mL of toluene, add 1 mmol of p-toluenesulfonic acid, place in a thermostat at 50°C for 10 h, separate and purify the obtained product to obtain modified phytosterol;

[0039] (2) Weigh 0.8 g of soybean phosphatidylcholine and 0.2 g of the modified phytosterol prepared in step (1) and dissolve them in 30 mL of a chloroform-methanol mixture with a volume ratio of 1:1, place them in a dark place at 25° C. for 15 min, remove the chloroform-methanol mixture by rotary evaporation, add 50 mL of phosphate buffered saline and 0.1 g of Bifidobacterium longum powder, the pH value of the phosphate buffered saline is 7, and perform ultrasonic treatment in a constant temperature water bath at 40° C. for 30 min at an ultrasonic power of 200 W to obtain probiotic liposomes;

[0040] (3) Weigh 5 g of modified chitosan and dissolve it in 50 mL of water, add 0.5 g of the probiotic liposomes prepared in step (2), stir in a constant temperature water bath at 40°C at a speed of 100 rpm, perform ultrasonic treatment for 10 min at a power of 200 W, and freeze-dry to obtain a composite probiotic liposome freeze-dried powder.

[0041] The present embodiment provides a method for preparing a high-bioactivity goat colostrum composite freeze-dried powder, which specifically comprises the following steps: adding composite probiotic liposome freeze-dried powder and oligomannosaccharide to goat colostrum powder by weight, stirring evenly, and obtaining a high-bioactivity goat colostrum composite freeze-dried powder.

[0042] Example 2

[0043] A high-bioactive sheep colostrum composite freeze-dried powder comprises the following components in parts by weight: 70 parts of sheep colostrum powder, 8 parts of composite probiotic liposome freeze-dried powder, and 9 parts of oligomannosaccharides.

[0044] The preparation method of modified chitosan comprises the following steps:

[0045] S1, weigh 3g of chloroacetic acid, add it to 30mL of isopropanol, and stir until it is completely dissolved to obtain a chloroacetic acid-isopropanol solution;

[0046] S2. Weigh 3 g of chitosan and add it to 20 mL of isopropanol. Stir for 2 h to obtain a mixed solution. Then add 30 mL of a 30% sodium hydroxide solution by mass, stir in a constant temperature water bath at 60°C at a speed of 200 rpm, slowly add 30 mL of the chloroacetic acid-isopropanol solution prepared in step S1, react for 3 h, adjust the pH value to 7, wash, and dry to obtain modified chitosan.

[0047] The preparation method of the composite probiotic liposome freeze-dried powder comprises the following steps:

[0048] (1) Weigh 10 mmol of gallic acid and 10 mmol of phytosterol and dissolve them in 50 mL of tetrahydrofuran, add 1 mmol of p-toluenesulfonic acid, place in a thermostat at 50°C for 20 h, separate and purify the obtained product to obtain modified phytosterol;

[0049] (2) Weigh 1 g of soybean phosphatidylcholine and 0.4 g of the modified phytosterol prepared in step (1) and dissolve them in 30 mL of a chloroform-methanol mixture with a volume ratio of 1:1, place them in a dark place at 25° C. for 30 min, remove the chloroform-methanol mixture by rotary evaporation, add 60 mL of phosphate buffered saline and 0.1 g of Lactobacillus acidophilus powder, the pH value of the phosphate buffered saline is 7, and perform ultrasonic treatment in a constant temperature water bath at 50° C. for 40 min at an ultrasonic power of 200 W to obtain probiotic liposomes;

[0050] (3) Weigh 7.5 g of modified chitosan and dissolve it in 50 mL of water, add 0.75 g of the probiotic liposomes prepared in step (2), stir in a constant temperature water bath at 50°C at a speed of 200 rpm, perform ultrasonic treatment for 20 min at a power of 200 W, and freeze-dry to obtain a composite probiotic liposome freeze-dried powder.

[0051] The present embodiment provides a method for preparing a high-bioactivity goat colostrum composite freeze-dried powder, which specifically comprises the following steps: adding composite probiotic liposome freeze-dried powder and oligomannosaccharide to goat colostrum powder by weight, stirring evenly, and obtaining a high-bioactivity goat colostrum composite freeze-dried powder.

[0052] Example 3

[0053] A high-bioactive sheep colostrum composite freeze-dried powder comprises the following components in parts by weight: 65 parts of sheep colostrum powder, 7 parts of composite probiotic liposome freeze-dried powder, and 8 parts of oligomannosaccharides.

[0054] The preparation method of modified chitosan comprises the following steps:

[0055] S1, weigh 3g of chloroacetic acid, add it to 30mL of isopropanol, and stir until it is completely dissolved to obtain a chloroacetic acid-isopropanol solution;

[0056] S2. Weigh 2.5 g of chitosan and add it to 20 mL of isopropanol. Stir for 1.5 h to obtain a mixed solution. Then add 30 mL of a 30% sodium hydroxide solution by mass, stir in a constant temperature water bath at 55°C at a speed of 150 rpm, slowly add 25 mL of the chloroacetic acid-isopropanol solution prepared in step S1, react for 2.5 h, adjust the pH value to 7, wash, and dry to obtain modified chitosan.

[0057] The preparation method of the composite probiotic liposome freeze-dried powder comprises the following steps:

[0058] (1) Weigh 10 mmol of gallic acid and 10 mmol of phytosterol and dissolve them in 50 mL of toluene, add 1 mmol of p-toluenesulfonic acid, place in a thermostat at 50°C for 15 h, separate and purify the obtained product to obtain modified phytosterol;

[0059] (2) Weigh 0.9 g of soybean phosphatidylcholine and 0.3 g of the modified phytosterol prepared in step (1) and dissolve them in 30 mL of a chloroform-methanol mixture with a volume ratio of 1:1, place them in a dark place at 25° C. for 20 min, remove the chloroform-methanol mixture by rotary evaporation, add 55 mL of phosphate buffered saline and 0.1 g of Lactobacillus acidophilus powder, the pH value of the phosphate buffered saline is 7, and perform ultrasonic treatment in a constant temperature water bath at 45° C. for 35 min at an ultrasonic power of 200 W to obtain probiotic liposomes;

[0060] (3) Weigh 7 g of modified chitosan and dissolve it in 50 mL of water, add 0.7 g of the probiotic liposomes prepared in step (2), stir in a constant temperature water bath at 45°C at a speed of 150 rpm, perform ultrasonic treatment for 15 min at a power of 200 W, and freeze-dry to obtain a composite probiotic liposome freeze-dried powder.

[0061] The present embodiment provides a method for preparing a high-bioactivity goat colostrum composite freeze-dried powder, which specifically comprises the following steps: adding composite probiotic liposome freeze-dried powder and oligomannosaccharide to goat colostrum powder by weight, stirring evenly, and obtaining a high-bioactivity goat colostrum composite freeze-dried powder.

[0062] Comparative Example 1

[0063] This comparative example provides a composite probiotic liposome freeze-dried powder, which is different from Example 1 in that modified phytosterols are not prepared, and phytosterols are used instead of modified phytosterols. The remaining components and component contents are the same as those in Example 1.

[0064] Comparative Example 2

[0065] This comparative example provides a composite probiotic liposome freeze-dried powder, which is different from Example 1 in that the composite probiotic liposome freeze-dried powder does not contain modified chitosan, and the other components and component contents are the same as those in Example 1.

[0066] Experimental Example 1

[0067] The composite probiotic liposome freeze-dried powder prepared in Examples 1-3 of the present invention and Comparative Examples 1 and 2 was tested for encapsulation efficiency, and the specific method was as follows: the composite probiotic liposome freeze-dried powder was weighed and added to a mixture of methanol and distilled water, ultrasonicated at 400W for 10-20min, and then centrifuged at 8000r / min for 20min to obtain bacterial mud, phosphate buffer was added to the bacterial mud, lysozyme was added, enzymolysis was performed at 40°C for 60min, and 500W was ultrasonicated for 30min to obtain an enzymolysis solution, 1mL of the enzymolysis solution was taken, 5mL of Coomassie Brilliant Blue G-250 reagent was added, mixed and allowed to stand for 2min, and the absorbance was measured at 595nm of an ultraviolet spectrophotometer, and the liposome-encapsulated probiotic protein content A1 was calculated, and the total probiotic protein content A2 was determined using the same method, and the encapsulation efficiency = A1 / A2 × 100%.

[0068] Figure 1 The encapsulation efficiency test results of the composite probiotic liposome freeze-dried powder prepared in Examples 1-3 of the present invention and Comparative Examples 1 and 2 are shown in the figure. As shown in the figure, the encapsulation efficiency of Examples 1-3 is 85.3%, 86.1%, and 84.9%, respectively, and the encapsulation efficiency of Comparative Examples 1 and 2 is 77.3% and 68.4%, respectively. The encapsulation efficiency of Examples 1-3 is higher than that of Comparative Example 1, indicating that the encapsulation efficiency can be increased after the modified plant sterol introduces gallic acid. The encapsulation efficiency of Examples 1-3 is higher than that of Comparative Example 2, indicating that the addition of modified chitosan increases the encapsulation efficiency and improves the protective effect of the composite probiotic liposome freeze-dried powder on probiotics.

[0069] Experimental Example 2

[0070] The storage stability test of the composite probiotic liposome freeze-dried powder prepared in Examples 1-3 of the present invention and Comparative Examples 1 and 2 is performed. The specific method is as follows: the composite probiotic liposome freeze-dried powder is weighed and placed in a constant temperature and humidity chamber at a temperature of 30° C. and a humidity of 60%. After storage for 60 days, the encapsulation efficiency test is performed with reference to Experimental Example 1.

[0071] Figure 2 The storage stability test results of the composite probiotic liposome freeze-dried powder prepared in Examples 1-3 and Comparative Examples 1 and 2 of the present invention are shown in the figure. As shown in the figure, the test results of Examples 1-3 are 82.8%, 83.7%, and 81.5%, respectively, and the test results of Comparative Examples 1 and 2 are 51.2% and 52.3%, respectively. Combining Experimental Example 1 and Experimental Example 2, the encapsulation rate of Examples 1-3 changes very little after storage for 60 days, and the decrease in Comparative Example 1 is more obvious, indicating that the modified plant sterols can effectively improve the storage stability of the composite probiotic liposome freeze-dried powder after introducing gallic acid. The decrease in Comparative Example 2 is larger than that in Examples 1-3, indicating that the coating of modified chitosan can improve the storage stability of the composite probiotic liposome freeze-dried powder.

[0072] Experimental Example 3

[0073] The composite probiotic liposome freeze-dried powder prepared in Examples 1-3 of the present invention and Comparative Examples 1 and 2 was subjected to a simulated gastric juice test, and the specific method was as follows: 3 g of pepsin was dissolved in 1 L of a 0.5% NaCl solution, and the pH was adjusted to 2 to prepare artificial gastric juice, and the composite probiotic liposome freeze-dried powder prepared in Examples 1-3 and Comparative Examples 1-2 was added to the artificial gastric juice, according to the gastric emptying time of 4 h, under 37° C. water bath conditions, and shaken at a speed of 100 rpm, and the probiotic content was measured at 0 h and 4 h, respectively. The content at 0 h was taken as the reference value, and 4 h was taken as the final value. The preservation rate was calculated by the following formula: preservation rate = final value / reference value × 100%.

[0074] Figure 3The results of the simulated gastric juice test of the composite probiotic liposome freeze-dried powder prepared in Examples 1-3 of the present invention and Comparative Examples 1 and 2 are shown in the figure. As shown in the figure, the preservation rates of the probiotics in Examples 1-3 and Comparative Examples 1 and 2 are 92.2%, 91.4%, 92.6%, 82.7% and 68.4%, respectively; the preservation rate of the probiotics in Examples 1-3 is significantly higher than that in Comparative Example 2, indicating that the coating of modified chitosan reduces the damage of gastric juice to the probiotics and plays a protective role.

[0075] Experimental Example 4

[0076] The composite probiotic liposome freeze-dried powder prepared in Examples 1-3 of the present invention was subjected to a simulated digestion test, and the specific method was as follows: 10 g of trypsin and 3 g of bovine bile salt were dissolved in 1 L of a 0.5% NaCl solution, and the pH was adjusted to 7.5 to prepare artificial intestinal fluid. The composite probiotic liposome freeze-dried powder prepared in Examples 1-3 was first subjected to a simulated gastric fluid experiment, and then added to the artificial intestinal fluid, and shaken at 37 ° C and 100 rpm for 6 h to simulate the intestinal digestion process. The probiotic content was measured at 0 h and 6 h, respectively. The content at 0 h was taken as the reference value, and 6 h was taken as the final value. The release rate of the probiotics was calculated by the following formula: Release rate = (reference value-final value) / reference value × 100%.

[0077] Figure 4 This is a simulated digestion test result diagram of the composite probiotic liposome freeze-dried powder prepared in Examples 1-3 of the present invention. As shown in the figure, the release rates of Examples 1-3 are 87.6%, 88.1%, and 86.9%, respectively; the release rates of the probiotics in Examples 1-3 are all greater than 85%, indicating that the probiotics have a good release effect in the intestine.

[0078] Experimental Example 5

[0079] The redispersibility test of the composite probiotic liposome freeze-dried powder prepared in Examples 1-3 of the present invention was carried out, and the specific method was as follows: the composite probiotic liposome freeze-dried powder prepared in Examples 1-3 was added to deionized water at 40° C. in an amount of 0.2 g / ml, dissolved by unidirectional stirring at 10 rpm, filtered through a 60-mesh sieve, and the dry weight of the filtered material was taken as the agglomerate mass, the mass of the composite probiotic liposome freeze-dried powder added to the solution was taken as the total mass, and the agglomeration rate was calculated by the following formula; agglomeration rate = agglomeration mass / total mass × 100%.

[0080] Figure 5 This is a result graph of the redispersibility test of the composite probiotic liposome freeze-dried powder prepared in Examples 1-3 of the present invention. As shown in the figure, the agglomeration rates of Examples 1-3 are 17.2%, 16.9%, and 16.3%, respectively, indicating that the composite probiotic liposome freeze-dried powder has good redispersibility.

[0081] Experimental Example 6

[0082] After the composite probiotic liposome freeze-dried powder prepared in Example 1 was reconstituted, its microstructure was observed using a transmission electron microscope.

[0083] Figure 6 This is a transmission electron micrograph of the composite probiotic liposome freeze-dried powder prepared in Example 1 of the present invention. As can be seen from the figure, the freeze-dried powder has a round appearance, a smooth surface, and uniform size after being reconstituted.

[0084] While the embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that many changes, modifications, substitutions and variations can be made to the embodiments without departing from the principles and spirit of the invention.

[0085] The present invention and its implementation methods are described above, which is not restrictive. The drawings are only one of the implementation methods of the present invention, and the actual application is not limited thereto. In short, if ordinary technicians in the field are inspired by it and design methods and embodiments similar to the technical solution without creativity without departing from the purpose of the invention, they should all fall within the protection scope of the present invention.

Claims

1. A highly biologically active sheep colostrum composite freeze-dried powder, characterized in that: The high-bioactive sheep colostrum composite freeze-dried powder comprises the following components in parts by weight: 60-70 parts of sheep colostrum powder, 5-8 parts of composite probiotic liposome freeze-dried powder, and 6-9 parts of oligomannosaccharide; the composite probiotic liposome freeze-dried powder is prepared from the following components: gallic acid, phytosterol, soybean phosphatidylcholine, modified chitosan, and probiotic powder; the probiotic powder is Bifidobacterium longum powder or Lactobacillus acidophilus powder; The preparation method of the modified chitosan comprises the following steps: S1, weigh chloroacetic acid and add it to isopropanol, the amount of chloroacetic acid added to isopropanol is 100 mg / mL, and stir until completely dissolved to obtain a chloroacetic acid-isopropanol solution; S2, weigh chitosan and add it to isopropanol, stir for 1-2h to obtain a mixed solution, then add sodium hydroxide solution, stir in a constant temperature water bath, slowly add the chloroacetic acid-isopropanol solution prepared in step S1, react for 2-3h, adjust the pH value to 7, wash, and dry to obtain modified chitosan; The preparation method of the composite probiotic liposome freeze-dried powder comprises the following steps: (1) Weighing gallic acid and phytosterols, dissolving them in a reaction solvent, adding a catalyst, placing the mixture in a thermostat at 50° C. for 10-20 hours, and separating and purifying the obtained products to obtain modified phytosterols; (2) weighing soybean phosphatidylcholine and the modified plant sterol prepared in step (1) and dissolving them in an organic solvent, hydrating them in the dark at 25° C. for 15-30 minutes, removing the organic solvent by rotary evaporation, adding phosphate buffered saline solution and probiotic powder, and ultrasonically treating them in a constant temperature water bath to obtain probiotic liposomes; (3) Weigh the modified chitosan and dissolve it in water, add the probiotic liposomes prepared in step (2), stir in a constant temperature water bath, perform ultrasonic treatment, and freeze-dry to obtain a composite probiotic liposome freeze-dried powder.

2. The highly biologically active goat colostrum composite freeze-dried powder according to claim 1, characterized in that: In step S2, the amount of chitosan added to the isopropanol is 100-150 mg / mL, the volume ratio of the mixed solution to the sodium hydroxide solution is 4:5, and the volume ratio of the mixed solution to the chloroacetic acid-isopropanol solution is 1:1-1.

5.

3. The highly biologically active goat colostrum composite freeze-dried powder according to claim 1, characterized in that: In step S2, the sodium hydroxide solution is a solution of sodium hydroxide in water, the mass fraction of the sodium hydroxide solution is 30%, the stirring temperature of the constant temperature water bath is 50-60° C., and the stirring speed is 100-200 rpm.

4. The method for preparing the highly biologically active goat colostrum composite freeze-dried powder according to claim 1, characterized in that: In step (1), the molar ratio of the gallic acid to the phytosterol is 1:

1.

5. The highly biologically active goat colostrum composite freeze-dried powder according to claim 1, characterized in that: In step (1), the catalyst is p-toluenesulfonic acid, the molar ratio of the catalyst to phytosterol is 1:0.1, and the reaction solvent is toluene or tetrahydrofuran.

6. The highly biologically active goat colostrum composite freeze-dried powder according to claim 1, characterized in that: In step (2), the weight ratio of the probiotic powder, the soybean phosphatidylcholine and the modified plant sterol is 1:8-10:2-4, the temperature of the constant temperature water bath is 40-50°C, the time of the ultrasonic treatment is 30-40 min, the power is 200 W, the volume ratio of the phosphate buffered saline solution to the organic solvent is 5-6:3, and the pH value of the phosphate buffered saline solution is 7.

7. The highly biologically active goat colostrum composite freeze-dried powder according to claim 1, characterized in that: In step (3), the amount of modified chitosan added to water is 100-150 mg / mL, the mass ratio of modified chitosan to probiotic liposome is 10:1, the stirring temperature of the constant temperature water bath is 40-50°C, the stirring speed is 100-200 rpm, the ultrasonic treatment time is 10-20 min, and the power is 200 W.

8. A method for preparing the highly biologically active goat colostrum composite freeze-dried powder according to claim 1, characterized in that: The method specifically comprises the following steps: adding compound probiotic liposome freeze-dried powder and oligomannosaccharide into goat colostrum powder, stirring evenly, and obtaining compound freeze-dried goat colostrum powder with high biological activity.

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