Prebiotic microcapsule as well as preparation method and application thereof

Abstract

The invention belongs to the field of prebiotic microcapsules, and discloses a prebiotic microcapsule as well as a preparation method and application thereof. The preparation method comprises the following steps: mixing the hydrolyzed glutinous rice flour, the tea polyphenol, the L-glutamine, the raffinose, the arabinoxylan, the mannan oligosaccharide, the curcumin and the sodium alginate to obtain a water phase component I; mixing psyllium husk powder, konjac glucomannan, chitosan, guar gum and hot water to obtain a mixed system, performing pH regulation and cooling treatment, and mixing calcium chloride powder with the cooled mixed system until the calcium chloride powder is dissolved and saturated to obtain a water phase component II; mixing rapeseed oil, lecithin, optional olive oil and optional tea oil to prepare an oil phase component; and mixing and stirring the water-phase component I and the oil-phase component to obtain an oil-water mixed emulsion, mixing the oil-water mixed emulsion with the heated water-phase component II, curing, and centrifuging to obtain the prebiotic microcapsule. The prebiotic microcapsule disclosed by the invention can effectively solve the problem of flora imbalance of vagina micro-ecology.

Description

Technical Field

[0001] The present invention belongs to the field of prebiotic microcapsules, and more specifically, relates to a prebiotic microcapsule and a preparation method and application thereof. Background Art

[0002] Vaginitis is a common disease that troubles women's health and quality of life. There is currently no safe and effective intervention measure. Drug treatment is effective in the short term but is prone to recurrence. The occurrence of vaginal inflammation is closely related to pH imbalance, immune barrier damage and microecological imbalance in the vaginal microenvironment, especially the excessive proliferation of pro-inflammatory anaerobic bacteria such as Lactobacillus iners, which inhibits the growth of hydrogen peroxide-producing Lactobacillus crispatus and destroys the antibacterial effect mediated by hydrogen peroxide, which is the key to vaginal bacterial growth and bacterial vaginitis. Therefore, the preparation of prebiotic microcapsules with improved vaginal microecological environment is the key to the preparation of drugs and disinfectant preparations for the prevention and treatment of vaginal inflammation. Summary of the invention

[0003] The purpose of the present invention is to address the deficiencies of the prior art and to provide a prebiotic microcapsule and a preparation method and application thereof. The prebiotic microcapsule of the present invention can effectively solve the problem of flora imbalance in vaginal microecology.

[0004] In order to achieve the above object, the first aspect of the present invention provides a method for preparing prebiotic microcapsules, the preparation method comprising the following steps:

[0005] S1: mixing and stirring the hydrolyzed glutinous rice flour, tea polyphenols, L-glutamine, raffinose, arabinoxylan, manno-oligosaccharide, curcumin and sodium alginate to obtain an aqueous phase component 1;

[0006] S2: mixing and stirring psyllium husk powder, konjac gum, chitosan, guar gum and hot water to obtain a mixed system; adjusting the pH of the mixed system and cooling the mixed system, mixing calcium chloride powder with the mixed system after the cooling treatment until the calcium chloride powder is saturated, and taking the supernatant to obtain a second aqueous phase component;

[0007] S3: mixing rapeseed oil, lecithin, optional olive oil and optional tea oil to prepare an oil phase component;

[0008] S4: mixing and stirring the water phase component 1 with the oil phase component to obtain an oil-water mixed emulsion; mixing the heated water phase component 2 with the oil-water mixed emulsion and performing a static solidification treatment to obtain a solidified system; centrifuging the water layer of the solidified system to obtain the prebiotic microcapsules.

[0009] According to the present invention, preferably, in step S1, the preparation method of the hydrolyzed glutinous rice flour is acid hydrolysis or enzyme hydrolysis;

[0010] Preferably, the acid hydrolysis method comprises: uniformly mixing glutinous rice flour and citric acid aqueous solution, heating in a water bath to obtain an acid hydrolyzate; uniformly mixing the cooled acid hydrolyzate with sodium citrate to neutralize the acidity to obtain the hydrolyzed glutinous rice flour.

[0011] Preferably, the enzymatic hydrolysis method comprises: adding glutinous rice flour to an amylase solution for enzymatic hydrolysis, heating and boiling in a water bath to terminate the enzymatic hydrolysis, and cooling to obtain the hydrolyzed glutinous rice flour.

[0012] According to the present invention, preferably, the concentration of the citric acid aqueous solution is 0.40-0.60 g / L; the usage ratio of glutinous rice flour to citric acid aqueous solution is 20-80 g glutinous rice flour / L citric acid aqueous solution;

[0013] The water bath heating temperature is 60-100°C and the time is 20-60min;

[0014] The temperature of the cooled acid hydrolyzate is 18-30°C;

[0015] The final pH of the hydrolyzed glutinous rice flour after neutralization of acidity is 5.5-7.5;

[0016] According to the present invention, preferably, the amylase solution is food-grade non-thermostable α-amylase, 100 U / L;

[0017] The usage ratio of glutinous rice flour to amylase solution is 20-80g glutinous rice flour / L amylase solution;

[0018] The temperature of the water bath is 25-35℃, and the time is 10-30min; boil for 5-10min and cool to 18-30℃.

[0019] According to the present invention, preferably, the mass ratio of the glutinous rice flour to tea polyphenols, L-glutamine, raffinose, arabinoxylan, manno-oligosaccharide, curcumin and sodium alginate is 5-20: 0.1-1.5: 0.8-1.2: 3-5: 1-2: 1-3: 1.5-2.5: 4.5-5.5.

[0020] According to the present invention, preferably, in step S2, the ratio of hot water to the total mass of the psyllium husk powder, konjac gum, chitosan and guar gum is 80-200:1;

[0021] The mass ratio of the psyllium husk powder, konjac gum, chitosan and guar gum is 0.8-1.2: 0.5-2.5: 0.5-2.5: 0.5-2.5;

[0022] The temperature of the hot water is 95-100°C;

[0023] The temperature of the mixed system after the cooling treatment is 35-50° C., and the pH is 6.0-7.5.

[0024] According to the present invention, preferably, in step S3, the mass ratio of rapeseed oil, lecithin, olive oil and tea oil is 25-50:0.5-2:0-20:0-20;

[0025] Preferably, the mass ratio of the rapeseed oil, lecithin, olive oil and tea oil is 25-50:0.8-1.2:5-15:5-15.

[0026] According to the present invention, preferably, in step S4:

[0027] The mixing time of the water phase component 1 and the oil phase component is 40-80 minutes;

[0028] The temperature of the heated aqueous phase component 2 is 35-55°C;

[0029] The oil-water mixed emulsion is added dropwise to the heated water phase component 2, and then allowed to stand for solidification. The diameter of the liquid outlet of the device used for the dropwise addition is 0.40-1.55 mm.

[0030] The curing time is 0.5-2h;

[0031] The rotation speed of the centrifugal treatment is 1000-5000 rpm, and the time is 5-20 min.

[0032] In the present invention, as a preferred embodiment, in step S4, the water phase component is added dropwise into the oil phase component under stirring with a magnetic stirrer at room temperature, and stirred at high speed for 60 minutes to form an oil-water mixed emulsion.

[0033] In the present invention, the oil layer of the solidification system is removed by a method known to those skilled in the art. As a preferred embodiment, the oil layer of the solidification system is removed by absorption.

[0034] The second aspect of the present invention provides prebiotic microcapsules prepared by the preparation method of the prebiotic microcapsules.

[0035] The third aspect of the present invention provides the use of the prebiotic microcapsules in the preparation of medicines, medical devices and disinfectants for preventing and / or treating vaginal inflammation.

[0036] The beneficial effects of the technical solution of the present invention are as follows:

[0037] The microcapsules prepared by the method of the present invention can effectively inhibit the growth of Lactobacillus iners, Gardnerella vaginalis and Prevotellatimonensis, and promote the growth of Lactobacillus crispatus, thereby achieving the effect of regulating the balance of vaginal microecology.

[0038] Other features and advantages of the present invention will be described in detail in the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] The above and other objects, features and advantages of the present invention will become more apparent through a more detailed description of exemplary embodiments of the present invention with reference to the accompanying drawings.

[0040] Figure 1 The effect of the prebiotic microcapsules prepared by the preparation method of the prebiotic microcapsules provided in Example 1 of the present invention is shown (*, p<0.05; **, p<0.01; ***, p<0.001; OD600 ， The absorbance value of visible light at a wavelength of 600 nanometers; "Lactobacillus crispatus" Lactobacillus crispatus, "Lactobacillus iners" Lactobacillus iners, "Gardnerella vaginalis" Gardnerella vaginalis). DETAILED DESCRIPTION

[0041] The preferred embodiments of the present invention will be described in more detail below. Although the preferred embodiments of the present invention are described below, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided to make the present invention more thorough and complete, and to fully convey the scope of the present invention to those skilled in the art.

[0042] Example 1

[0043] This embodiment provides a method for preparing prebiotic microcapsules, the preparation method comprising the following steps:

[0044] S1: glutinous rice flour and citric acid aqueous solution (0.5 g / L) are mixed uniformly at a dosage ratio of 1:10 g / L, and heated in a water bath at 80°C for 30 min to obtain an acid hydrolyzate; the acid hydrolyzate cooled to room temperature 25°C is mixed uniformly with sodium citrate to obtain hydrolyzed glutinous rice flour (pH 6);

[0045] The hydrolyzed glutinous rice flour, tea polyphenols, L-glutamine, raffinose, arabinoxylan, manno-oligosaccharide, curcumin and sodium alginate are mixed and stirred uniformly to obtain a water phase component 1;

[0046] The mass ratio of the glutinous rice flour to tea polyphenols, L-glutamine, raffinose, arabinoxylan, manno-oligosaccharide, curcumin and sodium alginate is 10:1:1:3:1:2:2:5.

[0047] S2: Mix and stir psyllium husk powder, konjac gum, chitosan, guar gum and 100° C. hot water to obtain a mixed system, adjust the pH to 5.8, cool to 50° C., mix calcium chloride powder with the cooled mixed system until the calcium chloride powder is saturated, and take the supernatant to obtain a second aqueous phase component;

[0048] The mass ratio of hot water to the total mass of the psyllium husk powder, konjac gum, chitosan and guar gum is 100:1; the mass ratio of the psyllium husk powder, konjac gum, chitosan and guar gum is 1:2:2:2.

[0049] S3: mixing rapeseed oil, lecithin, olive oil and tea oil to prepare an oil phase component;

[0050] The mass ratio of the rapeseed oil, lecithin, olive oil and tea oil is 30:1:10:10.

[0051] S4: Add the water phase component 1 drop by drop into the oil phase component under stirring with a magnetic stirrer at room temperature of 25°C, and stir at high speed for 60 minutes to form an oil-water mixed emulsion. After heating the water phase component 2 to 50°C, use a 0.67mm needle to drop the oil-water mixed emulsion into the water phase component 2, and solidify for one hour to obtain a solidified system. Absorb the oil layer of the solidified system, and centrifuge the water layer of the solidified system at 3000 rpm for 15 minutes to obtain the prebiotic microcapsules.

[0052] The present invention uses MRS medium to culture Lactobacillus iners and Lactobacillus crispatus under anaerobic conditions, and uses NYCIII broth medium to culture Gardnerella vaginalis and Prevotella timonensis. The initial OD600 of each bacterium under the initial culture conditions is 0.1. The microcapsules prepared in Example 1 are added to each culture medium at a ratio of 1 g / L, and the control is not added with microcapsules. After culturing for 48 hours, the OD600 is detected to analyze the changes in the abundance of a single bacterium (each group has three replicates). Figure 1 As shown, it can be seen that after adding the microcapsules prepared in Example 1, Lactobacillus crispatus proliferated significantly, while the growth of the main pathogens of bacterial vaginitis, Lactobacillus iners, Gardnerella vaginalis and Prevotella timonensis were significantly inhibited.

[0053] The embodiments of the present invention have been described above, and the above description is exemplary, not exhaustive, and is not limited to the disclosed embodiments. Many modifications and changes will be apparent to those skilled in the art without departing from the scope and spirit of the described embodiments.

Claims

1. A method for preparing prebiotic microcapsules, characterized in that: The preparation method comprises the following steps: S1: mixing and stirring the hydrolyzed glutinous rice flour, tea polyphenols, L-glutamine, raffinose, arabinoxylan, manno-oligosaccharide, curcumin and sodium alginate to obtain an aqueous phase component 1; S2: mixing and stirring psyllium husk powder, konjac gum, chitosan, guar gum and hot water to obtain a mixed system; adjusting the pH of the mixed system and cooling the mixed system, mixing calcium chloride powder with the mixed system after the cooling treatment until the calcium chloride powder is saturated, and taking the supernatant to obtain a second aqueous phase component; S3: mixing rapeseed oil, lecithin, optional olive oil and optional tea oil to prepare an oil phase component; S4: mixing and stirring the water phase component 1 with the oil phase component to obtain an oil-water mixed emulsion; mixing the heated water phase component 2 with the oil-water mixed emulsion and performing a static solidification treatment to obtain a solidified system; centrifuging the water layer of the solidified system to obtain the prebiotic microcapsules.

2. The method for preparing prebiotic microcapsules according to claim 1, wherein: In step S1, the preparation method of the hydrolyzed glutinous rice flour is acid hydrolysis or enzyme hydrolysis; Preferably, the acid hydrolysis method comprises: uniformly mixing glutinous rice flour and citric acid aqueous solution, heating in a water bath to obtain an acid hydrolyzate; uniformly mixing the cooled acid hydrolyzate with sodium citrate to neutralize the acidity to obtain the hydrolyzed glutinous rice flour. Preferably, the enzymatic hydrolysis method comprises: adding glutinous rice flour to an amylase solution for enzymatic hydrolysis, heating and boiling in a water bath to terminate the enzymatic hydrolysis, and cooling to obtain the hydrolyzed glutinous rice flour.

3. The method for preparing prebiotic microcapsules according to claim 2, wherein: The concentration of the citric acid aqueous solution is 0.40-0.60 g / L; the usage ratio of glutinous rice flour to citric acid aqueous solution is 20-80 g glutinous rice flour / L citric acid aqueous solution; The water bath heating temperature is 60-100°C and the time is 20-60min; The temperature of the cooled acid hydrolyzate is 18-30°C; The final pH of the hydrolyzed glutinous rice flour after neutralization of acidity is 5.5-7.

5.

4. The method for preparing prebiotic microcapsules according to claim 2, wherein: The amylase solution is food-grade non-thermostable α-amylase, 100U / L; The usage ratio of glutinous rice flour to amylase solution is 20-80g glutinous rice flour / L amylase solution; The temperature of the water bath is 25-35℃, and the time is 10-30min; boil for 5-10min and cool to 18-30℃.

5. The method for preparing prebiotic microcapsules according to claim 2, wherein: The mass ratio of the glutinous rice flour to tea polyphenols, L-glutamine, raffinose, arabinoxylan, mannooligosaccharide, curcumin and sodium alginate is 5-20: 0.1-1.5：0.8-1.2：3-5：1-2：1-3：1.5-2.5：4.5-5.5。 6. The method for preparing prebiotic microcapsules according to claim 1, wherein: In step S2, the ratio of hot water to the total mass of the psyllium husk powder, konjac gum, chitosan and guar gum is 80-200:1; The mass ratio of the psyllium husk powder, konjac gum, chitosan and guar gum is 0.8-1.2: 0.5-2.5: 0.5-2.5: 0.5-2.5; The temperature of the hot water is 95-100°C; The temperature of the mixed system after the cooling treatment is 35-50° C., and the pH is 6.0-7.

5.

7. The method for preparing prebiotic microcapsules according to claim 1, wherein: In step S3, the mass ratio of rapeseed oil, lecithin, olive oil and tea oil is 25-50:0.5-2:0-20:0-20; Preferably, the mass ratio of rapeseed oil, lecithin, olive oil and tea oil is 25-50: 0.8-1.2：5-15：5-15。 8. The method for preparing prebiotic microcapsules according to claim 1, wherein: In step S4: The mixing time of the water phase component 1 and the oil phase component is 40-80 minutes; The temperature of the heated aqueous phase component 2 is 35-55°C; The oil-water mixed emulsion is added dropwise to the heated water phase component 2, and then allowed to stand for solidification. The diameter of the liquid outlet of the device used for the dropwise addition is 0.40-1.55 mm. The curing time is 0.5-2h; The rotation speed of the centrifugal treatment is 1000-5000 rpm, and the time is 5-20 min.

9. The prebiotic microcapsules prepared by the method for preparing prebiotic microcapsules according to any one of claims 1 to 8.

10. Use of the prebiotic microcapsules according to claim 9 in the preparation of medicines, medical devices and disinfectants for preventing and / or treating vaginal inflammation.

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