A method for preparing a composition having an immune function enhancing effect and use thereof

Abstract

This invention discloses a method for preparing a composition with immune-enhancing effects and its application, belonging to the field of health product technology. The preparation method is as follows: Astragalus membranaceus, Lycium barbarum, and Prunus cerasifera are ground into powder to obtain premix A; Cordyceps militaris and Pleurotus ostreatus are washed, vacuum freeze-dried, and pulverized to obtain premix B; Lemon myrtle leaves are cleaned, dried, and pulverized to obtain lemon myrtle leaf powder; Premix A, Premix B, and lemon myrtle leaf powder are mixed evenly, and deionized water is added and mixed thoroughly to obtain a fermentation substrate; a mixed bacterial agent is added to the fermentation substrate for fermentation; after fermentation, the fermentation liquid is refrigerated and sterilized, the supernatant is collected after centrifugation and filtered, and then the filtered fermentation liquid is mixed evenly with concentrated apple juice and yeast β-glucan to obtain a composition with immune-enhancing effects. The composition of this invention can effectively activate the activity of immune cells in the body, enhance the body's resistance to pathogens, and improve the market competitiveness of the product.

Description

Technical Field

[0001] This invention relates to the field of health product technology, and more specifically, to a method for preparing a composition that enhances immune function and its application. Background Technology

[0002] Immunity is a crucial barrier for the human body to resist the invasion of external pathogens and maintain normal physiological functions. With the fast pace of life, increased stress, and unhealthy lifestyle habits, the number of people with weakened immunity is rising, leading to many sub-health problems such as physical weakness, lethargy, and fatigue. Existing immune-enhancing products, especially traditional Chinese medicine products, often use traditional pulverizing and decoction processes, resulting in insufficient dissolution of active ingredients and poor absorption by the body, failing to fully exert their immune-enhancing effects. Some commercially available products add hormones and chemical enhancers to achieve better results, posing safety risks with long-term use and failing to meet consumers' demand for natural, safe, and highly effective immune-enhancing products. Therefore, developing a novel immune-enhancing composition has significant market value and application prospects. Summary of the Invention

[0003] The purpose of this invention is to overcome the shortcomings of the prior art and provide a method for preparing a composition with immune-enhancing effects and its application.

[0004] To achieve the above objectives, the present invention adopts the following technical solution:

[0005] A method for preparing a composition with immune-enhancing effects and its application, comprising the following steps:

[0006] (a) Grind Astragalus membranaceus, Lycium barbarum and Prunus armeniaca into powder to obtain premix A; wash Cordyceps militaris and Pleurotus ostreatus, freeze dry under vacuum, and pulverize to obtain premix B; clean, dry and pulverize Lemon Peach Leaf to obtain Lemon Peach Leaf Powder.

[0007] (b) Mix premix A, premix B and lemon myrtle leaf powder evenly, add deionized water and mix well to obtain fermentation substrate;

[0008] (c) Add the mixed microbial agent to the fermentation substrate for fermentation;

[0009] (d) After fermentation, the fermentation broth is refrigerated and sterilized. After centrifugation, the supernatant is filtered and then the filtered fermentation broth is mixed evenly with concentrated apple juice and yeast β-glucan to obtain a composition that enhances immune function.

[0010] The present invention is further configured such that, in step (a), the mass ratio of Astragalus membranaceus to Lycium barbarum, Ficus hirta, Cordyceps militaris, Pleurotus ostreatus, and Prunus cerasifera leaves is (25-35):(18-22.4):(12.5-17):(16-20.2):(8-15):(4-7.6).

[0011] The present invention is further configured such that, in step (b), the ratio of the mixed powder of premix A, premix B, and lemon myrtle leaf powder to deionized water is 1g:8-15mL.

[0012] The present invention is further configured such that, in step (c), the mixed microbial agent is a composition of *Lactobacillus plantarum*, *Lactobacillus fermentum*, and *Lactococcus lactis* subsp. *lactococcus*.

[0013] The present invention is further configured such that the mass ratio of *Lactobacillus plantarum* to *Lactobacillus fermentum* and *Lactococcus lactis* subsp. *lactococcus* is (1-4):(1.5-3):1.

[0014] The present invention is further configured such that, in step (c), the mixed microbial agent accounts for 2-5% of the mass of the fermentation substrate.

[0015] The present invention is further configured such that, in step (c), the fermentation time is 48-72 hours.

[0016] The present invention is further configured such that, in step (d), the mass ratio of the filtered fermentation broth to concentrated apple juice and yeast β-glucan is (100-200):(6-10):1.

[0017] On the other hand, the present invention provides the application of the above preparation method in the preparation of a composition with enhanced immune function.

[0018] In summary, the present invention has the following beneficial effects:

[0019] This product is prepared by fermentation using Astragalus membranaceus, Lycium barbarum, Prunus armeniaca, Cordyceps militaris, Pleurotus ostreatus, and Lemon senna leaves. The complementary and synergistic effects of each ingredient form a multi-pathway immune regulation mechanism, which can effectively activate the activity of immune cells, increase the secretion level of immune factors, and significantly enhance the body's resistance to pathogens, thus ensuring product efficacy and enhancing its market competitiveness. Detailed Implementation

[0020] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0021] Example 1

[0022] (a) Prepare 30 parts by weight of Astragalus membranaceus, 20.4 parts by weight of Lycium barbarum, 15 parts by weight of Ficus hirta, 18.1 parts by weight of Cordyceps militaris, 12 parts by weight of Mushroom roe, and 6.2 parts by weight of Lemon myrtle leaves. Grind Astragalus membranaceus, Lycium barbarum, and Ficus hirta to 100 mesh to obtain premix A; wash Cordyceps militaris and Mushroom roe and freeze-dry them under vacuum (vacuum degree is absolute pressure 10 Pa, temperature -30℃, drying time 24h), and grind them to 100 mesh to obtain premix B; clean Lemon myrtle leaves and dry them with hot air at 40-50℃ to constant weight, and grind them to 100 mesh to obtain Lemon myrtle leaf powder.

[0023] (b) Mix premix A, premix B and lemon myrtle leaf powder evenly, add deionized water (the ratio of mixed powder to deionized water is 1g:10mL) and stir evenly to obtain fermentation substrate.

[0024] (c) Add the mixed microbial agent (the mass ratio of Lactobacillus plantarum, Lactobacillus fermentum, and Lactococcus lactis subsp. lactis in the mixed microbial agent is 2:2:1) to the fermentation substrate (the mixed microbial agent accounts for 3% of the mass of the fermentation substrate), stir evenly, and place it in a constant temperature incubator at 37℃ for 72h. Stir once every 6h during the fermentation process (stirring time is 5min).

[0025] (d) After fermentation, the fermentation liquid was placed at 4℃ for 30 min, sterilized, and centrifuged (3000 r / min for 15 min). The supernatant was filtered through a 0.22 μm filter membrane to obtain a clear fermentation liquid. The fermentation liquid was mixed evenly with concentrated apple juice and yeast β-glucan (the mass ratio of fermentation liquid to concentrated apple juice and yeast β-glucan was 120:8:1). After washing, peeling and cored the apples, they were cut into small pieces, crushed and juiced, and then filtered through a coarse filter and a 200-mesh filter. The filtrate was concentrated to a sugar content of 20 Birx to obtain concentrated apple juice. A composition with immune-enhancing effects was obtained.

[0026] Comparative Example 1

[0027] The composition for enhancing immune function was prepared according to Example 1, except that no five-finger peach was added.

[0028] Comparative Example 2

[0029] The composition for enhancing immune function was prepared according to Example 1, except that egg yolk mushrooms were not added.

[0030] Comparative Example 3

[0031] The composition for enhancing immune function was prepared according to Example 1, except that lemon myrtle leaves were not added.

[0032] The immune-improving effects of the compositions in Example 1 and the comparative examples were tested using the following methods:

[0033] (1) Effects on NK cell activity

[0034] One hundred Balb / C mice (weighing 18-22g) were randomly divided into 5 groups (n=20 per group, half male and half female): blank control group, Example 1 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group. Each group of mice was administered the corresponding composition by gavage (twice daily, morning and evening, 0.2mL / 10g body weight each time; the blank control group was administered an equal volume of physiological saline). After 30 days of continuous administration, the mice were sacrificed, and the spleen was aseptically removed, shredded, passed through a 200-mesh sieve, washed three times with Hank's solution, and prepared with complete 1640 culture medium to a concentration of 5×10⁻⁶. 6 Cell suspension per mL. 300 μL of cell suspension from each mouse was aliquoted into 96-well plates (100 μL per well), and target cells (YAC-1 cells, 1 × 10⁶ cells) were added to each well. 5 NK cell density was measured at 100 μL / ml. Simultaneously, 8 wells were prepared for natural release (100 μL target cells + 100 μL culture medium) and 8 wells for maximum release (100 μL target cells + 100 μL 1% NP-40). The cells were incubated at 37°C and 5% CO2 for 4 hours, then centrifuged at 1500 rpm for 5 minutes. 100 μL of the supernatant from each well was transferred to another culture plate, and 100 μL of basal medium was added to each well. After 10 minutes, 30 μL of 1 mol / L HCl solution was added to terminate the reaction. The OD value was measured at 490 nm, and the NK cell viability was calculated.

[0035] (2) Mouse carbon clearance test

[0036] One hundred Balb / C mice (weighing 18-22g) were randomly divided into 5 groups (n=20 per group, half male and half female): blank control group, Example 1 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group. Each group of mice was administered the corresponding composition by gavage (twice daily, morning and evening, 0.2mL / 10g body weight each time; the blank control group was administered an equal volume of physiological saline). After 14 consecutive days of administration, 30 minutes after the last administration, each mouse was injected with Indian ink via the tail vein at a dose of 0.1mL / 10g body weight. 20μL of blood was collected from the orbital vein at 1 min and 5 min, respectively, and added to 2mL of 0.1% NaCO3 solution. The absorbance (OD) was measured at 680nm using a spectrophotometer, and the clearance index (K) was calculated.

[0037] One hundred ICR mice (weighing 20-25g) were randomly divided into 5 groups (n=20 per group, half male and half female): blank control group, Example 1 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group. Each group of mice was administered the corresponding composition by gavage (twice daily, morning and evening, 0.2mL / 10g body weight each time; the blank control group was administered an equal volume of physiological saline). After 30 days of continuous administration, all mice were intraperitoneally injected with 40mg / kg sodium pentobarbital to induce sleep. The sleep latency and sleep duration of each group were recorded during the experiment.

[0038] The test results of the effects of the compositions of Example 1 and the comparative examples on immunity and sleep are shown in the table below:

[0039]

[0040] The material ratios and preparation conditions of the composition for enhancing immune function in this invention are not limited to those described in Example 1. In step (a), the mass ratio of Astragalus membranaceus to Lycium barbarum, Ficus hirta, Cordyceps militaris, Lemon yolk mushroom, and lemon myrtle leaf can be adjusted to (25-35):(18-22.4):(12.5-17):(16-20.2):(8-15):(4-7.6); in step (b), the mixed powder of premix A, premix B, and lemon myrtle leaf powder is mixed with deionized water. The material-to-liquid ratio can be adjusted to 1g:8-15mL; in step (c), the mass ratio of *Lactobacillus plantarum* to *Lactobacillus fermentatus* and *Lactococcus lactis* subsp. *lactococcus* can be adjusted to (1-4):(1.5-3):1, the mass ratio of the mixed inoculum to the fermentation substrate can be adjusted to 2-5%, and the fermentation time can be adjusted to 48-72h; in step (d), the mass ratio of the filtered fermentation broth to concentrated apple juice and yeast β-glucan can be adjusted to (100-200):(6-10):1.

[0041] The above description is merely a preferred embodiment of the present invention. The scope of protection of the present invention is not limited to the above embodiments. All technical solutions falling within the scope of the present invention's concept are within the scope of protection of the present invention. It should be noted that for those skilled in the art, any improvements and modifications made without departing from the principles of the present invention should also be considered within the scope of protection of the present invention.

Claims

1. A method for preparing a composition with immune-enhancing effects, characterized in that, Includes the following steps: (a) Grind Astragalus membranaceus, Lycium barbarum and Prunus armeniaca into powder to obtain premix A; wash Cordyceps militaris and Pleurotus ostreatus, freeze dry under vacuum, and pulverize to obtain premix B; clean, dry and pulverize Lemon Peach Leaf to obtain Lemon Peach Leaf Powder. (b) Mix premix A, premix B and lemon myrtle leaf powder evenly, add deionized water and mix well to obtain fermentation substrate; (c) Add the mixed microbial agent to the fermentation substrate for fermentation; (d) After fermentation, the fermentation broth is refrigerated and sterilized. After centrifugation, the supernatant is filtered and then the filtered fermentation broth is mixed evenly with concentrated apple juice and yeast β-glucan to obtain a composition that enhances immune function.

2. The method for preparing a composition with immune-enhancing effects according to claim 1, characterized in that, In step (a), the mass ratio of Astragalus membranaceus to Lycium barbarum, Ficus hirta, Cordyceps militaris, Trichosanthes kirilowii, and Prunus cerasifera leaves is (25-35): (18-22.4): (12.5-17): (16-20.2): (8-15): (4-7.6).

3. The method for preparing a composition with immune-enhancing effects according to claim 1, characterized in that, In step (b), the ratio of the mixed powder of premix A, premix B, and lemon myrtle leaf powder to deionized water is 1g:8-15mL.

4. The method for preparing a composition with immune-enhancing effects according to claim 1, characterized in that, In step (c), the mixed microbial agent is a combination of *Lactobacillus plantarum*, *Lactobacillus fermentum*, and *Lactococcus lactis* subsp. *lactococcus*.

5. The method for preparing a composition with immune-enhancing effects according to claim 4, characterized in that, The mass ratio of *Lactobacillus plantarum* to *Lactobacillus fermentum* and *Lactococcus lactis* subsp. *lactococcus* was (1-4):(1.5-3):

1.

6. The method for preparing a composition with immune-enhancing function according to claim 1, characterized in that, In step (c), the mixed microbial agent accounts for 2-5% of the mass of the fermentation substrate.

7. The method for preparing a composition with immune-enhancing effects according to claim 1, characterized in that, In step (c), the fermentation time is 48-72 hours.

8. The method for preparing a composition with immune-enhancing function according to claim 1, characterized in that, In step (d), the mass ratio of the filtered fermentation broth to concentrated apple juice and yeast β-glucan is (100-200):(6-10):

1.

9. The use of the preparation method according to any one of claims 1-8 in the preparation of an immune-enhancing composition.

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