Seaweed gel microspheres, preparation method thereof and application of seaweed gel microspheres in covering bitter taste of ganoderma lucidum aqueous extract

Abstract

The invention discloses seaweed gel microspheres, a preparation method thereof and application of the seaweed gel microspheres in covering bitter taste of a ganoderma lucidum aqueous extract, and the preparation method comprises the following steps: (1) adding ganoderma lucidum aqueous extract powder into water, stirring and dissolving to prepare a ganoderma lucidum aqueous extract solution; (2) mixing the lucid ganoderma aqueous extract solution prepared in the step (1) with a sodium alginate aqueous solution, stirring for 10-60 minutes, and then homogenizing; and (3) spraying the solution obtained after homogenization in the step (2) into a mixed solution of chitosan oligosaccharide and calcium chloride, filtering by using rapid filter paper to obtain generated microspheres, and freeze-drying to obtain the seaweed gel microspheres. By constructing the composite microspheres of sodium alginate and chitosan oligosaccharide / calcium and combining with a freeze-drying process, the ganoderma lucidum water extract can be embedded, the bitter taste of the ganoderma lucidum water extract is effectively covered, and the effect of targeted intestinal slow release is achieved.

Description

Technical Field

[0001] The present invention relates to the technical field of food or health products, and particularly relates to an alginate microsphere, a preparation method thereof, and an application for masking the bitterness of Ganoderma lucidum aqueous extract. Background Art

[0002] Many bitter substances not only endow foods with bitterness, but also have physiological activities such as antioxidant, anti-tumor, blood pressure lowering, and blood sugar lowering. The main bitter-tasting substances are plant nutrients such as phenols, flavonoids, isoflavonoids, terpenes, alkaloids, and bitter peptides. Ganoderma lucidum is a traditional precious Chinese medicine in China, with the effects of replenishing middle qi, tonifying and strengthening the body, strengthening the healthy qi and consolidating the constitution, and prolonging life, and is listed as a precious medicine. Ganoderma lucidum contains functional components such as polysaccharides, triterpenoids, proteins, sterols, and trace elements. Triterpenoids are the main source of the bitterness of Ganoderma lucidum. At present, more than 130 Ganoderma triterpenoids have been extracted from Ganoderma lucidum, which are mainly divided into tetracyclic lanostane-type triterpenoids and pentacyclic triterpenoids. According to different structures and functional groups, Ganoderma triterpenoids are further divided into ganoderic acids, ganospore acids, ganoderic acid lactones, methyl ganoderates, ganodermanontriol, and ganoderaldehyde. The aqueous extract of Ganoderma lucidum is rich in various activities, such as having auxiliary protection against radiation hazards, anti-tumor, enhancing immunity, anti-aging, detoxifying, and antiviral effects. However, due to its rich triterpenoids, it also brings a relatively high degree of bitterness, resulting in an unpleasant taste experience.

[0003] Embedding can physically or chemically encapsulate the bitter substances of Ganoderma lucidum inside the wall material, delay the release of its bitterness, effectively inhibit the perception of bitterness, while effectively retaining its active ingredients and extending its shelf life. Microencapsulation technology is a relatively simple and effective method for masking the unpleasant odor of drugs through the embedding effect. Different microcapsule wall materials not only play a role in masking the taste, but also can achieve the effects of slow release and targeted release. Embedding can play a role in masking bitterness, but the process technology still needs to be continuously improved. Moreover, the embedding technology for masking the bitterness of the aqueous extract of Ganoderma lucidum remains to be studied. Summary of the Invention

[0004] In view of the above problems existing in the prior art, the present invention provides an alginate microsphere, a preparation method thereof, and an application for masking the bitterness of Ganoderma lucidum aqueous extract. By constructing a composite microsphere of sodium alginate and chitosan oligosaccharide / calcium and combining with the freeze-drying process, the present invention can encapsulate the aqueous extract of Ganoderma lucidum, effectively mask the bitterness of the aqueous extract of Ganoderma lucidum, and achieve the effect of targeted intestinal slow release.

[0005] The technical solution of the present invention is as follows:

[0006] The first object of the present invention is to provide a method for constructing an alginate microsphere, and the method comprises the following steps:

[0007] (1) Add the Ganoderma lucidum water extract powder to water, stir to dissolve, and obtain the Ganoderma lucidum water extract solution;

[0008] (2) Mix the Ganoderma lucidum water extract solution prepared in step (1) with the sodium alginate aqueous solution, stir for 10 - 60 min, and then perform homogenization treatment;

[0009] (3) Spray the solution obtained after homogenization in step (2) into the mixed solution of chitosan oligosaccharide and calcium chloride, filter out the generated microspheres with a rapid filter paper, and perform freeze - drying to obtain the alginate microspheres.

[0010] In one embodiment of the present invention, in step (1), the concentration of the Ganoderma lucidum water extract solution is 1 - 5 wt%.

[0011] In one embodiment of the present invention, in step (1), the concentration of the Ganoderma lucidum water extract solution is 3 wt%.

[0012] In one embodiment of the present invention, in step (1), the extraction method of the Ganoderma lucidum water extract powder is: add 10 - 20 times the weight of water to the Ganoderma lucidum fruiting body, extract 1 - 3 times, each extraction for 1 - 3 h, and concentrate to a relative density of 1.01 - 1.25.

[0013] In one embodiment of the present invention, in step (2), the concentration of the sodium alginate aqueous solution is 1 - 4 wt%.

[0014] In one embodiment of the present invention, in step (2), the concentration of the sodium alginate aqueous solution is 3 wt%.

[0015] In one embodiment of the present invention, in step (2), the preparation method of the sodium alginate aqueous solution is: add sodium alginate to water, heat and stir at 50 - 100 °C for 25 - 60 min, and perform ultrasonic defoaming for 20 - 40 min to obtain the sodium alginate aqueous solution.

[0016] In one embodiment of the present invention, in step (2), the mass ratio of the Ganoderma lucidum water extract solution to the sodium alginate aqueous solution is 1:1 - 9.

[0017] In one embodiment of the present invention, in step (2), the homogenization pressure is 800 - 1500 bar, and it is repeated 2 - 3 times.

[0018] In one embodiment of the present invention, in step (3), in the mixed solution of chitosan oligosaccharide and calcium chloride, the concentration of chitosan oligosaccharide is 0.5 - 5 wt%, and the concentration of calcium chloride is 0.1 - 5 wt%.

[0019] In one embodiment of the present invention, in step (3), in the mixed solution of chitosan oligosaccharide and calcium chloride, the concentration of chitosan oligosaccharide is 1 wt%.

[0020] In one embodiment of the present invention, in step (3), in the mixed solution of chitosan oligosaccharide and calcium chloride, the mass ratio of chitosan oligosaccharide to calcium chloride is 1:0.5.

[0021] In one embodiment of the present invention, in step (3), spraying is carried out using a sprayer with a nozzle diameter of 0.1 - 0.3 mm, and the droplet diameter of the sprayed liquid is less than 1 mm.

[0022] Preferably, the nozzle diameter is 0.2 mm.

[0023] In the solution obtained after homogenization, the mass ratio of the water extract of Ganoderma lucidum, sodium alginate, chitosan oligosaccharide, and calcium chloride is 1:1:1:0.5.

[0024] In one embodiment of the present invention, in step (3), the conditions for freeze-drying are as follows: using a freeze dryer, the cold trap temperature is -57.4 °C, and the vacuum degree is 282 Pa. The second object of the present invention is to provide a sodium alginate microsphere prepared by the above method.

[0025] The third object of the present invention is to provide an application of the above sodium alginate microsphere for embedding the water extract of Ganoderma lucidum, masking bitterness, and achieving targeted intestinal slow release.

[0026] The beneficial technical effects of the present invention are as follows:

[0027] The present invention uses the construction of sodium alginate microspheres for embedding to mask the bitterness of the water extract of Ganoderma lucidum and slow down the release of bitter substances. Specifically, the loading capacity can reach 83%, the bitterness perception value is reduced by 5 times, and the results of the electronic tongue show that the relative taste intensity of bitterness is reduced by 3 times.

[0028] The participation of chitosan oligosaccharide in the present invention can increase the loading capacity of the water extract of Ganoderma lucidum. Compared with the microspheres without adding chitosan oligosaccharide, the loading capacity is increased to 1.12 times, and the stability of the sodium alginate microspheres is enhanced, which is beneficial for storage and improves the efficiency of masking bitterness, reducing the bitterness perception to half of the original.

[0029] The preparation process of the present invention only needs to mix, homogenize, and dry the bitter substance and the wall material substance, which is simpler, more efficient, and easier to promote compared with other embedding methods. Description of the Drawings

[0030] Figure 1 For testing the loading capacity of the Ganoderma lucidum embedding powder in Test Example 1, where the abscissa represents the ratio of the water extract of Ganoderma lucidum to sodium alginate, and abc represents data differences.

[0031] Figure 2 For the sensory evaluation results of the Ganoderma lucidum embedding powder embedding the water extract of Ganoderma lucidum in Test Example 1, where abc represents data differences.

[0032] Figure 3To test the TDA analysis results of Ganoderma lucidum embedded powder embedding Ganoderma lucidum aqueous extract in Test Example 1, where ab represents data difference.

[0033] Figure 4 For the electronic tongue analysis results of Ganoderma lucidum embedded powder in Test Example 1.

[0034] Figure 5 For the simulated digestion results of Ganoderma lucidum embedded powder in the oral cavity in Test Example 1, where abcdef represents data difference.

[0035] Figure 6 For the simulated digestion results of Cladosiphon okamuranus Tokida embedded powder in gastric juice in Test Example 1, where abcde represents data difference.

[0036] Figure 7 For the simulated digestion results of Ganoderma lucidum embedded powder in intestinal juice in Test Example 1, where abcdef represents data difference.

[0037] Figure 8 For the loading capacity of Ganoderma lucidum embedded powder in Test Example 2, where the abscissa represents the ratio of Ganoderma lucidum aqueous extract, and ab represents data difference.

[0038] Figure 9 For the electronic tongue analysis results of Ganoderma lucidum embedded powder in Test Example 2.

[0039] Figure 10 For the simulated digestion results of Ganoderma lucidum embedded powder in the oral cavity in Test Example 2, where abcdef represents data difference.

[0040] Figure 11 For the simulated digestion results of Ganoderma lucidum embedded powder in gastric juice in Test Example 2, where abcde represents data difference.

[0041] Figure 12 For the simulated digestion results of Ganoderma lucidum embedded powder in intestinal juice in Test Example 2, where abcdef represents data difference. Detailed implementation mode

[0042] The present invention will be specifically described below in conjunction with the drawings and embodiments.

[0043] Ganoderma lucidum aqueous extract is a composite component prepared from Ganoderma lucidum fruit bodies through the processes of water extraction, concentration and spray drying. Unless otherwise specified, the reagents used in the present invention are all purchased from commercial channels.

[0044] The following is an exemplary description: Sodium alginate is purchased from Qingdao Mingyue Seaweed Group Co., Ltd.; Chitosan oligosaccharide is purchased from Shanghai Macklin Biochemical Co., Ltd.

[0045] Example 1

[0046] A method for constructing alginate microspheres, comprising the following steps:

[0047] (1) Add 3 g of sodium alginate to 100 mL of water, heat and stir at 80 °C for 30 min, and ultrasonically defoam for 30 min to obtain a 3% sodium alginate solution.

[0048] (2) Add 3 g of Ganoderma lucidum water extract powder to 100 mL of water, stir to dissolve, and obtain a 3% Ganoderma lucidum water extract solution.

[0049] (3) Mix the Ganoderma lucidum water extract solution and the sodium alginate aqueous solution in a mass ratio of 1:1 - 9, and stir for 30 min; homogenize the mixed solution, and the homogenization pressure is 1000 bar.

[0050] (4) Add 1 g of chitosan oligosaccharide and 0.5 g of CaCl 2 to 100 mL of water, stir evenly to prepare a chitosan oligosaccharide - CaCl 2 solution. Then spray the homogenized sodium alginate - Ganoderma lucidum water extract mixture into the chitosan oligosaccharide - CaCl 2 solution. Construct microspheres through the interaction between sodium alginate, chitosan oligosaccharide, and calcium ions. Filter out the microspheres and place them in a laboratory freeze dryer for drying treatment. The specific parameters are a cold trap temperature of -57.4 °C and a vacuum degree of 282 Pa, which is the Ganoderma lucidum embedding powder.

[0051] Test Example 1:

[0052] (1) Determination of the loading amount of Ganoderma lucidum water extract

[0053] Weigh an appropriate amount of dry Ganoderma lucidum embedding powder into a beaker, add 1 g of NaHCO 3 and Na 3 C 6 H 5 O 7 ·2H 2 O as the cyst-breaking liquid, perform ultrasonic cyst-breaking treatment, then add 2 ml of 1 mol / L NaOH solution to make up to 50 mL, and take the filtered filtrate for dilution to measure the absorbance value. Calculate the loading amount of the microspheres through the standard curve equation.

[0054] Loading amount of Ganoderma lucidum water extract (%) = [measured Ganoderma lucidum water extract (g) / weight of embedding powder (g)] × 100%.

[0055] In the mixed solution of Ganoderma lucidum water extract and sodium alginate, different mass ratios of Ganoderma lucidum water extract to sodium alginate result in different loading amounts of Ganoderma lucidum water extract in the prepared Ganoderma lucidum embedding powder, as shown in Figure 1 Table 1. The loading amount of Ganoderma lucidum water extract in the Ganoderma lucidum embedding powder prepared under the conditions of Example 1 is more than 70%, and can even reach more than 80%.

[0056] Table 1

[0057] Ganoderma lucidum aqueous extract Sodium alginate Loading amount (%) 1 1 83±2.1 1 3 74±1.7 1 5 75±2.8 1 7 75±1.6 1 9 70±3.5

[0058] (2) Determination of taste

[0059] For the Ganoderma lucidum embedded powder prepared with the mass ratio of Ganoderma lucidum water extract to sodium alginate being 1:1 in Example 1, sensory evaluation and electronic tongue analysis were carried out. The specific operations are as follows:

[0060] Sensory evaluation was divided into the general Labeled Magnitude Scale (gLMS) scoring method and the Taste Dilution Analysis (TDA). 40 people (20 males and 20 females) were randomly selected. Before the specific experiment, behaviors that have a greater impact on taste, such as smoking, drinking alcohol, and chewing gum, were prohibited. The caffeine extract was set as the bitter reference substance, and the experiment was carried out in a professional sensory evaluation room.

[0061] For the gLMS scoring method, 5 mg of caffeine extract, Ganoderma lucidum water extract, and 10 mg of Ganoderma lucidum embedded powder were each given to the subjects for tasting, and scores were given after tasting. Evaluation criteria: 1.5 is hardly detectable, 6 is weak, 17 is moderate, 35 is strong, 52 is very strong, and 100 is the strongest. The results are shown in Figure 2 , the bitterness of the Ganoderma lucidum embedded powder is very low. For the TDA method, with the caffeine extract and Ganoderma lucidum water extract as references and distilled water as the blank, a 50 mg / mL Ganoderma lucidum embedded powder solution and 25 mg / mL caffeine extract and Ganoderma lucidum water extract solutions were prepared, and were gradually diluted in a 1:1 ratio. The bitterness was evaluated by triangular testing. In each test, two blank samples and one diluted sample were randomly labeled, and the test samples were randomly given to the subjects in ascending order of concentration, and the TD value was measured. The results are shown in Figure 3 , the TD value of the Ganoderma lucidum embedded powder is higher, indicating that the perception threshold of the Ganoderma lucidum embedded powder is lower, that is, it can be perceived at a lower concentration.

[0062] As a comparison, the Ganoderma lucidum water extract, caffeine extract, and Ganoderma lucidum water extract embedded under different conditions in this application; through such a comparison, under the embedding of the examples of this application, there is a better effect of masking bitterness.

[0063] Electronic tongue analysis was used to measure the taste of simulated oral fluid, coffee extract solution, Ganoderma lucidum water extract solution, and Ganoderma lucidum encapsulated powder solution using an electronic tongue. Before the test, the coffee extract solution, Ganoderma lucidum water extract solution, and Ganoderma lucidum encapsulated powder were diluted 1:1 with simulated salivary fluid (SSF) and stirred on a magnetic stirrer for 10 min to simulate the oral chewing process. The instrument was preheated for 20 min before the experiment, and then connected to a computer and the sensor signal amplification factor was set to 3 times. Pour each liquid to be tested (20 ml) into the electronic tongue beaker and number it. Another electronic tongue beaker was filled with deionized water as the cleaning solution, and the experiment was carried out in the order of cleaning solution, test solution, cleaning solution, and test solution. The sensor array was immersed in each sample 3 times for 140 seconds, and the mV output during the immersion was monitored. Each sample was measured 3 times. The taste analysis values of the electronic tongue are shown in Figure 4 , the caffeine extract showed relatively high intensities in bitterness and astringency, with a bitterness value reaching 14 and the astringency value also being significantly higher than that of other samples. This result is consistent with the known properties of caffeine, indicating that it may have a strong taste irritation. In contrast, the bitterness and astringency of the Ganoderma lucidum water extract were also relatively high, which may be due to the medicinal properties of Ganoderma lucidum. The Ganoderma lucidum encapsulated powder showed a significant decreasing trend compared with the two samples with stronger bitterness. In addition, through the analysis of the aftertaste (Aftertaste-A and Aftertaste-B), it was found that the aftertaste duration of the Ganoderma lucidum encapsulated powder was longer, which may be related to its encapsulation process. The encapsulation technology effectively delayed the release of bitter substances, thus prolonging the taste perception, which is consistent with the sensory evaluation results.

[0064] (3) Release of Ganoderma lucidum water extract in the encapsulated powder during simulated digestion

[0065] The release of Ganoderma lucidum water extract in the Ganoderma lucidum encapsulated powder prepared with a mass ratio of Ganoderma lucidum water extract to sodium alginate of 1:1 in Example 1 during simulated digestion was measured.

[0066] Determination of simulated oral release: Weigh 10 mg of Ganoderma lucidum encapsulated powder, add 10 ml of artificial simulated oral fluid at (37 ± 0.5) °C, place it in a constant temperature shaker (37 °C), and take the supernatant at 20, 30, 40, 50, 60, 120, and 300 s respectively, and measure its absorbance value at a wavelength of 271 nm. The cumulative release at each time point in the oral cavity is shown in Figure 5 , the release of Ganoderma lucidum encapsulated powder in the oral cavity was relatively low. After 3 minutes of oral digestion, the cumulative release of Ganoderma lucidum water extract was only 6.0%.

[0067] Determination of release performance in simulated gastric environment: The Ganoderma lucidum embedded powder after simulated oral digestion was centrifuged and filtered out, then added to 20 ml of artificial simulated gastric juice (37 °C). It was carried out under the conditions of a constant temperature shaker at 37 °C and 180 r / min. Samples were taken multiple times at 120 min, and the absorbance value was measured at a wavelength of 271 nm to further determine the concentration, and the cumulative release curve was plotted. The cumulative release degrees at each time point in the gastric juice are shown in Figure 6 , the release degree of the Ganoderma lucidum embedded powder in the stomach was also low. After 2 hours of gastric digestion, the cumulative release degree of the water extract of Ganoderma lucidum was only 9.11%.

[0068] Determination of release performance in simulated intestinal environment: The Ganoderma lucidum embedded powder after simulated gastric digestion was centrifuged and filtered out and placed in 40 ml of simulated intestinal fluid preheated at 37 °C, and then placed in a shaker (37 °C, 180 r / min). Samples were taken at 0.5, 1, 1.5, 2, 3, 4, 6, 8, and 12 h respectively, and the absorbance value was measured at a wavelength of 271 nm to determine the drug concentration in the sample solution. The cumulative release degrees at each time point in the intestinal fluid are shown in Figure 7 , after 12 hours of intestinal digestion of the Ganoderma lucidum embedded powder, the cumulative release degree of the water extract of Ganoderma lucidum reached 78.4%, indicating that the Ganoderma lucidum embedded powder has an intestinal targeting release effect.

[0069] According to the results of simulated digestion, the release of the embedded Ganoderma lucidum powder in the oral cavity is relatively slow, which indicates that embedding can reduce its release in the oral cavity, thereby reducing the binding of its bitter substances to receptors in the oral cavity and achieving the purpose of masking bitterness. By observing the cumulative release rate in the stomach and intestine, it can be found that more is released in the intestine after embedding, so as to achieve the purpose of targeting.

[0070] Example 2

[0071] A method for constructing alginate microspheres, comprising the following steps:

[0072] (1) Add 2 g of sodium alginate to 100 mL of water, heat and stir at 50 °C for 60 min, and defoam by ultrasound for 10 min to obtain a 2% sodium alginate solution;

[0073] (2) Add 1 g of the water extract powder of Ganoderma lucidum to 100 mL of water, stir and dissolve to obtain a 1% water extract solution of Ganoderma lucidum;

[0074] (3) Mix the water extract solution of Ganoderma lucidum and the aqueous sodium alginate solution in a mass ratio of 1:1 - 9, stir for 30 min; homogenize the mixed solution, and the homogenization pressure is 1000 bar;

[0075] (4) Add 0.5 g of chitosan oligosaccharide and 0.5 g of CaCl 2 to 100 mL of water, stir evenly to prepare chitosan oligosaccharide - CaCl 2Solution, and then the homogenized sodium alginate-Ganoderma lucidum aqueous extract mixture was sprayed into the chitosan oligosaccharide-CaCl 2 solution through the interaction between sodium alginate, chitosan oligosaccharide and calcium ions to construct microspheres. The microspheres were filtered out and dried in a laboratory freeze dryer with specific parameters: the cold trap temperature was -57.4 °C and the vacuum degree was 282 Pa, which was the Ganoderma lucidum embedding powder.

[0076] The loading amounts of the Ganoderma lucidum embedding powders prepared with different mass ratios of Ganoderma lucidum aqueous extract to sodium alginate are shown in Table 2.

[0077] Table 2

[0078] Ganoderma lucidum aqueous extract Sodium alginate Loading amount (%) 1 1 76±1.8 1 3 74±2.1 1 5 73±1.4 1 7 70±1.5 1 9 70±1.1

[0079] Comparative Example 1

[0080] A method for constructing alginate microspheres includes the following steps:

[0081] (1) Add 5 g of sodium alginate to 100 mL of water, heat and stir at 80 °C for 30 min, and ultrasonically defoam for 30 min to obtain a 5% sodium alginate solution;

[0082] (2) Add 3 g of Ganoderma lucidum aqueous extract powder to 100 mL of water, stir to dissolve to obtain a 3% Ganoderma lucidum aqueous extract solution;

[0083] (3) Mix the Ganoderma lucidum aqueous extract solution and the sodium alginate aqueous solution in a mass ratio of 1:1 - 9, stir for 30 min; homogenize the mixed solution, and the homogenization pressure is 1000 bar;

[0084] (4) Add 1 g of chitosan oligosaccharide and 0.5 g of CaCl 2 to 100 mL of water, stir evenly to prepare a chitosan oligosaccharide / CaCl 2 mixed solution;

[0085] Under this condition, sodium alginate has not been completely dissolved, and it is impossible to homogenize with Ganoderma lucidum aqueous extract into a solution with uniform composition, and thus it is impossible to combine with the chitosan oligosaccharide / CaCl 2 mixed solution by spraying, that is, it is impossible to form alginate microspheres.

[0086] Comparative Example 2

[0087] A method for constructing alginate microspheres includes the following steps:

[0088] (1) Add 3 g of sodium alginate to 100 mL of water, heat and stir at 80 °C for 30 min, and ultrasonically defoam for 30 min to obtain a 3% sodium alginate solution;

[0089] (2) Add 3 g of the water extract powder of Ganoderma lucidum to 100 mL of water, stir to dissolve, and obtain a Ganoderma lucidum water extract solution with a concentration of 3%.

[0090] (3) Mix the Ganoderma lucidum water extract solution and the sodium alginate aqueous solution in a mass ratio of 1:1 - 9, and stir for 30 min; homogenize the mixed solution, and the homogenization pressure is 1000 bar.

[0091] (4) Add 0.5 g of CaCl 2 to 100 mL of water, stir evenly, and then spray the homogenized sodium alginate-Ganoderma lucidum water extract mixture into the CaCl 2 solution through a 0.2 mm diameter atomizer. Construct microspheres through the interaction between sodium alginate and calcium ions, filter out the microspheres, and place them in a laboratory freeze dryer for drying treatment. The specific parameters are a cold trap temperature of -57.4 °C and a vacuum degree of 282 Pa. This is the Ganoderma lucidum embedding powder without chitosan oligosaccharide.

[0092] Test Example 2:

[0093] (1) Determination of the loading amount of Ganoderma lucidum water extract

[0094] Weigh an appropriate amount of the dried Ganoderma lucidum embedding powder without chitosan oligosaccharide into a beaker, add 1 g of NaHCO 3 and Na 3 C 6 H 5 O 7 ·2H 2 O as the cyst-breaking liquid, perform ultrasonic cyst-breaking treatment, then add 2 ml of 1 mol / L NaOH solution to make up the volume to 50 mL, and take the filtered filtrate for dilution to measure the absorbance value. Calculate the loading amount of the microspheres through the standard curve equation.

[0095] Loading amount of Ganoderma lucidum water extract (%) = [measured Ganoderma lucidum water extract (g) / weight of the embedding powder without chitosan oligosaccharide (g)] × 100%.

[0096] In the mixed solution of Ganoderma lucidum water extract and sodium alginate, different concentrations of sodium alginate result in different loading amounts of Ganoderma lucidum water extract in the Ganoderma lucidum embedding powder without chitosan oligosaccharide, as Figure 8 shown. The loading amount of Ganoderma lucidum water extract in the Ganoderma lucidum embedding powder without chitosan oligosaccharide prepared under the conditions of Comparative Example 2 is about 70%. Compared with the microspheres added with chitosan oligosaccharide, its loading amount of Ganoderma lucidum water extract is reduced, indicating that adding chitosan oligosaccharide can effectively improve the loading amount, that is, improve the embedding effect.

[0097] Table 3

[0098] Ganoderma lucidum aqueous extract Sodium alginate Loading amount (%) 1 1 74±1.7 1 3 73±1.4 1 5 71±1.3 1 7 72±2.8 1 9 68±2.2

[0099] (2) Determination of taste

[0100] The electronic tongue was used to analyze the Ganoderma lucidum embedded powder without chitosan oligosaccharide prepared from the Ganoderma lucidum aqueous extract solution and sodium alginate solution with a mass ratio of 1:1 in Comparative Example 2. The specific operation is as follows:

[0101] For the electronic tongue analysis, the electronic tongue was used to measure the taste of the Ganoderma lucidum embedded powder without chitosan oligosaccharide. Before the test, the sample was diluted 1:1 with simulated salivary fluid (SSF) and stirred on a magnetic stirrer for 10 min to simulate the oral chewing process. The instrument was preheated for 20 min before the experiment, and then connected to the computer and the sensor signal amplification factor was set to 3 times. Pour each liquid to be tested (20 ml) into the electronic tongue beaker and number it. Another electronic tongue beaker was filled with deionized water as the cleaning solution, and the experiment was carried out in the order of cleaning solution, test solution, cleaning solution, and test solution. The sensor array was immersed in each sample 3 times for 140 seconds, and the mV output during the immersion was monitored. The measurement was repeated 3 times. The taste analysis values of the electronic tongue are shown in Figure 9 , compared with the electronic tongue analysis of the microspheres prepared in Example 1, the intensity of the bitter taste perception increased after encapsulation, indicating that the Ganoderma lucidum embedded powder without chitosan oligosaccharide was relatively unstable in the simulated oral environment, resulting in the release of its bitter substances and being perceived by its receptor sensors.

[0102] (3) Release of the Ganoderma lucidum aqueous extract in the embedded powder during simulated digestion

[0103] The release of the Ganoderma lucidum aqueous extract in the Ganoderma lucidum embedded powder without chitosan oligosaccharide prepared from the Ganoderma lucidum aqueous extract solution and sodium alginate solution with a mass ratio of 1:1 in Comparative Example 2 during simulated digestion was measured.

[0104] Determination of the simulated oral release rate: Weigh 10 mg of the Ganoderma lucidum embedded powder without chitosan oligosaccharide, add 10 ml of artificial simulated oral fluid at (37 ± 0.5) °C, place it in a thermostatic shaker (37 °C), and take the supernatant at 20, 30, 40, 50, 60, 120, and 300 s respectively, and measure its absorbance value at a wavelength of 271 nm. The cumulative release rate at each time point in the oral cavity is shown in Figure 10 , compared with Example 1, after 3 minutes of oral digestion, its cumulative release rate in the oral cavity increased to 10.21%, indicating that the microspheres were relatively easily destroyed in the oral environment, leading to the release of bitter substances.

[0105] Determination of the release performance in the simulated gastric environment: The Ganoderma lucidum embedded powder without chitosan oligosaccharide after simulated oral digestion was centrifuged and filtered and added to 20 ml of artificial simulated gastric juice (37 °C), and carried out under the conditions of a thermostatic shaker at 37 °C and 180 r / min. Samples were taken multiple times at 120 min, and its absorbance value was measured at a wavelength of 271 nm to further determine the concentration, and the cumulative release curve was plotted. The cumulative release rate at each time point in the gastric juice is shown inFigure 11 , compared with Example 1, after 2 hours of gastric digestion, the cumulative release rate of the microspheres without chitosan oligosaccharide reached 16.22%, further illustrating its instability.

[0106] Determination of the release performance in the simulated intestinal environment: Centrifuge and filter out the Ganoderma lucidum embedding powder without chitosan oligosaccharide after simulated gastric digestion, put it into 40 ml of preheated simulated intestinal fluid at 37 °C, and then place it in a shaker (37 °C, 180 r / min). Samples are taken at 0.5, 1, 1.5, 2, 3, 4, 6, 8, and 12 h respectively, and the absorbance value is measured at a wavelength of 271 nm to determine the concentration of the water extract of Ganoderma lucidum in the sample solution. The cumulative release rate at each time point in the intestinal fluid is shown in Figure 12 , after 12 hours of intestinal digestion of the microspheres without chitosan oligosaccharide, the cumulative release rate of the water extract of Ganoderma lucidum was only 55.4%, indicating that the release amount of the microspheres without chitosan oligosaccharide in the intestine decreased, confirming its instability during digestion.

[0107] In summary, the addition of chitosan oligosaccharide is beneficial to improving the embedding efficiency, increasing the loading capacity, and better achieving the purpose of embedding. And the electronic tongue analysis shows that although the bitterness of the water extract of Ganoderma lucidum can still be effectively masked without adding chitosan oligosaccharide, its effect is far less than that of the sample after adding chitosan oligosaccharide. The simulated digestion results also confirm that the addition of chitosan oligosaccharide can effectively enhance the stability of the microspheres, reduce their release in the oral cavity, and reduce the bitter taste perception.

[0108] The embodiments provided above are not intended to limit the scope covered by the present invention, nor are the described steps intended to limit their execution order. Obvious improvements made by those skilled in the art to the present invention in combination with the existing common general knowledge also fall within the protection scope defined by the claims of the present invention.

Claims

1. A method for constructing alginate microspheres, characterized in that: The method comprises the following steps: (1) adding the Ganoderma lucidum water extract powder into water and stirring to dissolve, thereby preparing a Ganoderma lucidum water extract solution; (2) mixing the Ganoderma lucidum aqueous extract solution obtained in step (1) with the sodium alginate aqueous solution, stirring for 10-60 minutes, and then homogenizing; (3) spraying the homogenized solution obtained in step (2) into the mixed solution of chitosan oligosaccharide and calcium chloride, filtering out the generated microspheres with a rapid filter paper, and freeze-drying to obtain alginate microspheres.

2. The method according to claim 1, characterized in that In step (1), the concentration of the Ganoderma lucidum aqueous extract solution is 1-5wt%.

3. The method according to claim 1, characterized in that In step (1), the extraction method of the Ganoderma lucidum water extract powder is: adding water 10-20 times the weight of the Ganoderma lucidum fruiting body, extracting 1-3 times, each extraction time 1-3h, and concentrating to a relative density of 1.01-1.

25.

4. The method according to claim 1, characterized in that: In step (2), the concentration of the sodium alginate aqueous solution is 1-4 wt %.

5. The method according to claim 1, characterized in that In step (2), the sodium alginate aqueous solution is prepared by adding sodium alginate into water, heating and stirring at 50-100° C. for 25-60 min, and ultrasonically defoaming for 20-40 min to obtain the sodium alginate aqueous solution.

6. The method according to claim 1, characterized in that In step (2), the mass ratio of the Ganoderma lucidum aqueous extract solution to the sodium alginate aqueous solution is 1:1-9; and the homogenization pressure is 800-1500 bar.

7. The method according to claim 1, characterized in that In step (3), in the mixed solution of chitosan oligosaccharide and calcium chloride, the concentration of chitosan oligosaccharide is 0.5-5wt%, and the concentration of calcium chloride is 0.1-5wt%.

8. The method according to claim 1, characterized in that: In step (3), spraying is performed using a sprayer with a nozzle diameter of 0.1-0.3 mm and a spray droplet size of less than 1 mm.

9. Alginate microspheres prepared by the method according to any one of claims 1 to 8.

10. An application of the alginate microspheres according to claim 9, characterized in that: Used to encapsulate Ganoderma lucidum water extract, mask the bitter taste, and achieve targeted intestinal sustained release.

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