Shenshen cream capable of nourishing blood, tranquilizing mind, improving sleep quality and resisting oxidative damage and preparation method of Shenshen cream

Abstract

The invention belongs to the technical field of traditional Chinese medicines, and discloses a Shenshen cream with the effects of nourishing blood and tranquilizing mind, improving sleep and resisting oxidative damage. The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 10-16 parts of spina date seeds, 10-16 parts of red dates, 8-12 parts of longan aril, 8-12 parts of lotus seeds, 1-3 parts of radix angelicae sinensis, 5-10 parts of lucid ganoderma, 5-8 parts of radix astragali, 6-10 parts of chicory, 5-8 parts of lily bulbs and 3-5 parts of radix puerariae. According to the cream formula, the ganoderma lucidum and the angelica sinensis are fermented by adopting lactobacillus rhamnosus; and carrying out enzymolysis treatment on the astragalus membranaceus by using a compound enzyme. The ointment can significantly increase the number of red blood cells of a patient with blood deficiency; the sleep time is prolonged and the sleep quality is improved; the obvious oxidation resistance is realized. In addition, the ointment provided by the invention can improve the pain threshold value of mice in an acetic acid writhing method. Through unique raw material compatibility and process innovation, modern efficient utilization of traditional medicinal and edible materials is realized.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, specifically a Shishen San ointment that nourishes blood, calms the mind, improves sleep quality, and resists oxidative damage, along with its preparation method and application. Background Technology

[0002] With increasing life pressures and changing lifestyles in modern society, the number of people exhibiting blood deficiency and mental weakness syndrome is showing a significant upward trend. This syndrome is common in postpartum women, people with chronic diseases, and office workers under long-term high pressure. Symptoms most frequently include anemia, insomnia, palpitations, and anxiety. Its core pathogenesis lies in insufficient Qi and blood, and malnourishment of the mind and spirit. Clinical manifestations are complex and diverse, involving dysfunction of multiple systems. Current interventions for blood deficiency and mental weakness face the following prominent problems: First, Western medicine treatments are mostly symptomatic, such as using iron supplements to treat anemia and sedatives to improve insomnia, but they are difficult to fundamentally regulate the balance of Qi, blood, Yin, and Yang, and long-term use can easily lead to dependence. Second, while traditional Chinese medicine formulas have significant overall conditioning effects, they generally suffer from the following technical bottlenecks: single formulas have limited target points, making it difficult to fully cover the complex clinical manifestations of blood deficiency and mental weakness; traditional decoction processes have an extraction rate of less than 50% for key active ingredients such as Ganoderma lucidum polysaccharides and Angelica sinensis ferulic acid; and the dosage forms are limited, mostly decoctions or pills, which are inconvenient to take and have poor taste, resulting in low patient compliance. Therefore, there is an urgent need to develop a new type of preparation that is based on the theory of food and medicine sharing the same origin, uses modern preparation technology, and can simultaneously achieve multiple effects such as nourishing blood and heart, calming the mind and stabilizing the spirit, and regulating qi. Summary of the Invention

[0003] The purpose of this invention is to overcome the shortcomings of existing technologies and provide a medicinal and edible herbal paste that nourishes blood, calms the mind, improves sleep quality, and resists oxidative damage, along with its preparation and application methods, to solve the problems mentioned in the above-mentioned technical background. This invention relates to a medicinal and edible herbal paste and its preparation method. Its core research and development idea lies in combining modern biotechnology (such as microbial fermentation and compound enzymatic hydrolysis) with traditional Chinese medicine to improve the bioavailability of active ingredients in Chinese medicinal materials, achieving synergistic effects of nourishing blood, calming the mind, improving sleep, resisting oxidative damage, and relieving pain. This integration of modern biological knowledge with traditional Chinese medicine is a current hot trend. During the research and development process, this invention underwent extensive strain screening and enzyme combination optimization. Ultimately, Lactobacillus rhamnosus CICC6137 was selected to be combined with a cellulase-xylanase-β-glucosidase process to treat Ganoderma lucidum, Angelica sinensis, and Astragalus membranaceus. This specific combination was not obvious but rather an unexpected synergistic mechanism obtained through multiple iterative experiments (such as comparing strains like ATCC9595 with single enzyme hydrolysis). In the herbal matrix, the high enzyme activity of this strain and the targeted cell wall-breaking effect of the complex enzyme mutually enhance each other, triggering a multiplied release of polysaccharides and flavonoids and intestinal microbial remodeling. This results in an overall leap in the effects of nourishing blood and calming the mind, improving sleep, anti-oxidation, and analgesia. This unexpected synergistic effect is only manifested under this specific process, while conventional alternatives (as shown in the comparative example) only achieve partial effects, demonstrating the technical originality of this invention.

[0004] To achieve the above objectives, the present invention provides the following technical solution: In the first aspect, the present invention provides a medicinal and edible herbal paste that nourishes blood, calms the mind, improves sleep quality, and resists oxidative damage, composed of the following ingredients in the indicated weight proportions: 10-16 parts of jujube seed, 10-16 parts of red date, 8-12 parts of longan pulp, 8-12 parts of lotus seed, 1-3 parts of angelica, 5-10 parts of ganoderma, 5-8 parts of astragalus, 6-10 parts of chicory, 5-8 parts of lily bulb, and 3-5 parts of kudzu root.

[0005] One to three parts of Angelica sinensis and five to ten parts of Ganoderma lucidum were fermented with Lactobacillus rhamnosus CICC6137 and then mixed with the remaining ingredients in the formula. Five to eight portions of Astragalus membranaceus were subjected to enzymatic hydrolysis using a compound enzyme. In each 1,000 U of total enzyme activity of the compound enzyme, cellulase accounted for 500 U, xylanase for 300 U, and β-glucosidase for 200 U. After enzymatic hydrolysis, the enzyme was mixed with the remaining raw materials in the formula.

[0006] As a further option, the ingredients are composed of the following proportions: 13 parts jujube seed, 13 parts red date, 10 parts longan pulp, 10 parts lotus seed, 2 parts angelica, 7 parts lingzhi, 6 parts astragalus, 8 parts chicory, 6 parts lily bulb, and 4 parts kudzu root.

[0007] Secondly, this invention also provides a method for preparing a medicinal and edible herbal paste that nourishes blood, calms the mind, improves sleep quality, and resists oxidative damage, comprising the following steps: (1) Fermentation of Angelica sinensis and Ganoderma lucidum: 1) Preparation of Lactobacillus rhamnosus bacterial suspension: Take 100 mL of MRS liquid medium, sterilize at 121℃ for 15 minutes, and cool to 37℃; inoculate with 0.1 g of Lactobacillus rhamnosus CICC6137 lyophilized bacterial powder, and incubate at 37℃ for 24 hours to obtain Lactobacillus rhamnosus bacterial suspension with OD600≈1.0. Then inoculate it into MRS liquid medium at an inoculation rate of 2%, incubate at 37℃ until OD600≈1.5, and place it in a 4℃ refrigerator for later use.

[0008] 2) Inoculation: Grind 1-3 parts of Angelica sinensis and 5-10 parts of Ganoderma lucidum into powder and pass them through an 80-mesh sieve. Mix them evenly and inoculate with 3-5% Lactobacillus rhamnosus CICC6137 bacterial solution. Add distilled water to adjust the moisture content to 45%.

[0009] 3) Fermentation: Solid-state fermentation at 37℃ for 48-72 hours, stirring once every 12 hours during the period, maintaining pH 5.0-6.5. After fermentation, the bacterial strain in the fermentation product is inactivated with 3% H2O2 solution to obtain Angelica and Ganoderma fermentation product, which is then stored at 4℃ for later use.

[0010] (2) Enzymatic hydrolysis of Astragalus membranaceus: 5-8 parts of Astragalus membranaceus were pulverized and passed through a 60-mesh sieve. Phosphate buffer with pH 5.0-6.0 was added at a material-to-liquid ratio of 1:15. A compound enzyme was added at a total enzyme activity of 100 U / g Astragalus membranaceus (500 U of cellulase, 300 U of xylanase, and 200 U of β-glucosidase per 1,000 U of total enzyme activity). The mixture was enzymatically hydrolyzed in a constant temperature water bath at 50-55℃ for 2-4 hours with continuous stirring (200 rpm). The supernatant of the hydrolyzed Astragalus membranaceus was filtered and concentrated under reduced pressure at 60℃ (vacuum degree -0.09 MPa). The concentration was stopped when the relative density reached 1.15 at 60℃ using an online density meter. The hydrolysate of Astragalus membranaceus was obtained.

[0011] (3) Processing of other raw materials: 1) Sour jujube seed and longan pulp: 10-16 parts sour jujube seed and 8-12 parts longan pulp are dried at a low temperature of 45℃ and then pulverized and passed through a 200-mesh sieve; 2) Red dates, chicory, and kudzu root: 10-16 parts red dates, 6-10 parts chicory, and 3-5 parts kudzu root are dried at 65℃ and then pulverized through a 200-mesh sieve; 3) Lotus seeds and lily bulbs: Take 8-12 parts lotus seeds and 5-8 parts lily bulbs, mix them, add 10 times their weight of distilled water, and microwave extract twice (600W, 10 minutes / time). Combine the two extracts and concentrate them under reduced pressure at a vacuum degree of ≤-0.095 MPa. The endpoint is controlled at a relative density of 1.15 at 60℃ to obtain lotus seed and lily bulb concentrate.

[0012] (4) Extraction and concentration: Mix Angelica sinensis and Ganoderma lucidum fermentation product, Astragalus membranaceus enzymatic hydrolysate, Ziziphus jujuba seed powder, jujube powder, longan pulp powder, chicory powder and kudzu root powder, add 10 times the amount (about 2000 mL) of distilled water, reflux at 90℃ for 2 hours, filter, and collect the filtrate; mix the combined extract with lotus seed and lily bulb concentrate, and concentrate under reduced pressure at 60℃ to a relative density of 1.25.

[0013] (5) Refining into paste: Add 10% maltitol and 5% gum arabic as flavoring agent and stabilizer according to the weight of the paste, stir evenly, sterilize in water bath at 85°C for 15 minutes, fill into sterilized glass bottles while hot, seal, and the paste is obtained.

[0014] Thirdly, the present invention provides the application of the aforementioned medicinal and edible herbal paste in the preparation of products with functions of nourishing blood and calming the mind, improving sleep, resisting oxidative damage and relieving pain.

[0015] Compared with the prior art, the beneficial effects of the present invention are: 1. This invention utilizes Lactobacillus rhamnosus fermentation of Angelica sinensis and Ganoderma lucidum, combined with a compound enzymatic hydrolysis process for Astragalus membranaceus, to significantly improve the extraction rate and bioavailability of active ingredients. After fermentation, the polysaccharide yield of Ganoderma lucidum and Angelica sinensis increases by more than 40%, and the flavonoid yield reaches 90 mg / g, far exceeding the <50% dissolution rate of traditional water extraction processes. This demonstrates an unexpected synergistic effect in animal experiments.

[0016] 2. Synergistic effects: Angelica sinensis and Ganoderma lucidum fermented with Lactobacillus rhamnosus, and then combined with lily, have analgesic effects; the fermented products of Angelica sinensis and Ganoderma lucidum, combined with the enzymatic hydrolysate of Astragalus membranaceus, can synergistically increase the number of red blood cells, prolong sleep time and antioxidant function.

[0017] 3. During the research and development process, we also conducted detailed mechanistic speculation: For example, Lactobacillus rhamnosus CICC6137 fermentation of Ganoderma lucidum and Angelica sinensis may degrade the cell wall and transform active ingredients (such as Ganoderma lucidum triterpenes and Angelica sinensis ferulic acid) by producing extracellular polysaccharide enzymes, proteases and short-chain fatty acids (such as butyrate). These metabolites further regulate neurotransmitter pathways (such as activating GABA receptors and promoting inhibitory nerve conduction to improve sleep quality) and enhance the antioxidant system (such as activating the Nrf2 pathway and scavenging free radicals to alleviate oxidative damage). At the same time, it achieves blood-nourishing, calming and analgesic effects through gut microbiome remodeling (such as increasing the diversity of beneficial bacteria) (such as inhibiting inflammatory factors IL-6 and TNF-α). Attached Figure Description

[0018] Figure 1 This is a bar chart showing the effect of the traditional Chinese medicine and food homology paste of the present invention on the number of red blood cells in SD rats with blood deficiency syndrome; Figure 2This is a bar chart showing the effect of the traditional Chinese medicine and food homology paste of this invention on the sleep quality of mice with blood deficiency syndrome. Figure 3 This is a bar chart showing the effect of the traditional Chinese medicine and food homology paste of this invention on the DPPH free radical scavenging rate.

[0019] Figure 4 and Figure 5 This is a physical image of the herbal paste of the present invention. Detailed Implementation

[0020] The following specific examples illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that, unless otherwise specified, the following embodiments and features described therein can be combined with each other.

[0021] Angelica sinensis meets the 2020 Pharmacopoeia standards.

[0022] The Ganoderma lucidum meets the 2020 Pharmacopoeia standards.

[0023] Astragalus membranaceus meets the 2020 Pharmacopoeia standards.

[0024] Sour jujube seed meets the 2020 Pharmacopoeia standard.

[0025] Longan pulp meets the 2020 Pharmacopoeia standards.

[0026] Lotus seeds meet the 2020 Pharmacopoeia standards.

[0027] Chicory meets the 2020 Pharmacopoeia standards.

[0028] The lily meets the 2020 Pharmacopoeia standards.

[0029] Kudzu root meets the 2020 Pharmacopoeia standards.

[0030] Lactobacillus rhamnosus CICC6137 was purchased from the China Industrial Microbial Culture Collection Center.

[0031] Lactobacillus rhamnosus ATCC9595 was purchased from the China Industrial Microbial Culture Collection Center.

[0032] Cellulase, product number: 9012-54-8, purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0033] Xylanase, catalog number: 9025-57-4, purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0034] β-glucosidase, catalog number: 9001-22-3, purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0035] Example 1

[0036] A medicinal and edible paste, comprising the following ingredients by weight: 13 parts jujube seed, 13 parts red date, 10 parts longan pulp, 10 parts lotus seed, 2 parts angelica root, 7 parts ganoderma lucidum, 6 parts astragalus root, 8 parts chicory, 6 parts lily bulb, and 4 parts kudzu root. The ganoderma lucidum and angelica root are fermented with Lactobacillus rhamnosus to improve the bioavailability of their active ingredients. The astragalus root is enzymatically hydrolyzed using a compound enzyme composed of cellulase, xylanase, and β-glucosidase.

[0037] Example 2

[0038] A medicinal and edible paste, comprising the following ingredients by weight: 10 parts jujube seed, 16 parts red date, 8 parts longan pulp, 12 parts lotus seed, 1 part angelica root, 10 parts ganoderma lucidum, 5 parts astragalus root, 10 parts chicory, 5 parts lily bulb, and 5 parts kudzu root. The ganoderma lucidum and angelica root are fermented with Lactobacillus rhamnosus to improve the bioavailability of their active ingredients. The astragalus root is enzymatically hydrolyzed using a complex enzyme composed of cellulase, xylanase, and β-glucosidase.

[0039] Example 3

[0040] A medicinal and edible paste, comprising the following ingredients by weight: 16 parts jujube seed, 10 parts red date, 12 parts longan pulp, 8 parts lotus seed, 3 parts angelica root, 5 parts ganoderma lucidum, 8 parts astragalus root, 6 parts chicory, 8 parts lily bulb, and 3 parts kudzu root. The ganoderma lucidum and angelica root are fermented with Lactobacillus rhamnosus CICC6137 to improve the bioavailability of their active ingredients. The astragalus root is enzymatically hydrolyzed using a compound enzyme composed of cellulase, xylanase, and β-glucosidase.

[0041] Comparative Example 1 This comparative example is similar to Example 1, except that: Angelica sinensis is not fermented with Lactobacillus rhamnosus CICC6137.

[0042] Comparative Example 2 This embodiment is similar to Embodiment 1, except that the Ganoderma lucidum is not fermented by Lactobacillus rhamnosus CICC6137.

[0043] Comparative Example 3 This embodiment is similar to Embodiment 1, except that Angelica sinensis and Ganoderma lucidum are not fermented.

[0044] Comparative Example 4 This embodiment is similar to Embodiment 1, except that: Angelica sinensis and Ganoderma lucidum are fermented with Lactobacillus rhamnosus ATCC9595 instead of Lactobacillus rhamnosus CICC6137.

[0045] Comparative Example 5 This embodiment is similar to Embodiment 1, except that Astragalus membranaceus is not enzymatically hydrolyzed.

[0046] Comparative Example 6 This comparative example is similar to Example 1, except that: Astragalus membranaceus is not hydrolyzed by a complex enzyme (500 U of cellulase, 300 U of xylanase, and 200 U of β-glucosidase per 1,000 U of total enzyme activity), but is hydrolyzed by a complex enzyme (625 U of cellulase and 375 U of xylanase per 1,000 U of total enzyme activity).

[0047] Comparative Example 7 This comparative example is similar to Example 1, except that: Astragalus membranaceus is not hydrolyzed by a complex enzyme (500 U of cellulase, 300 U of xylanase, and 200 U of β-glucosidase per 1,000 U of total enzyme activity), but is hydrolyzed by a complex enzyme (600 U of xylanase and 400 U of β-glucosidase per 1,000 U of total enzyme activity).

[0048] Comparative Example 8 This comparative example is similar to Example 1, except that Astragalus membranaceus is not hydrolyzed by a complex enzyme (500 U of cellulase, 300 U of xylanase, and 200 U of β-glucosidase per 1,000 U of total enzyme activity), but is hydrolyzed by a complex enzyme (714 U of cellulase and 286 U of xylanase per 1,000 U of total enzyme activity).

[0049] Comparative Example 9 This comparative example is similar to Example 1, except that Angelica sinensis is omitted.

[0050] Comparative Example 10 This comparative example is similar to Example 1, except that Ganoderma lucidum is omitted.

[0051] Comparative Example 11 This comparative example is similar to Example 1, except that Astragalus membranaceus was removed.

[0052] Comparative Example 12 This comparative example is similar to Example 1, except that lilies are omitted.

[0053] Preparation method of Example 1: (1) Fermentation of Angelica sinensis and Ganoderma lucidum: 1) Preparation of Lactobacillus rhamnosus bacterial suspension: Take 100 mL of MRS liquid medium, sterilize at 121℃ for 15 minutes, and cool to 37℃; inoculate with 0.1 g of Lactobacillus rhamnosus CICC6137 lyophilized bacterial powder, and incubate at 37℃ for 24 hours to obtain Lactobacillus rhamnosus bacterial suspension with OD600≈1.0. Then inoculate it into MRS liquid medium at an inoculation rate of 2%, incubate at 37℃ until OD600≈1.5, and place it in a 4℃ refrigerator for later use.

[0054] 2) Inoculation: Grind 2 parts of Angelica sinensis and 7 parts of Ganoderma lucidum into powder and pass them through an 80-mesh sieve. Mix them evenly and inoculate with 3% Lactobacillus rhamnosus CICC6137 bacterial solution. Add distilled water to adjust the moisture content to 45%.

[0055] 3) Fermentation: Solid-state fermentation at 37℃ for 72 hours, stirring once every 12 hours during the period, maintaining pH 6.0±0.3. After fermentation, the bacterial strain in the fermentation product is inactivated with 3% H2O2 solution to obtain Angelica and Ganoderma fermentation product, which is then stored at 4℃ for later use.

[0056] (2) Enzymatic hydrolysis of Astragalus membranaceus: Six portions of Astragalus membranaceus were pulverized and passed through a 60-mesh sieve. Phosphate buffer with pH 5.8 was added at a material-to-liquid ratio of 1:15. A compound enzyme was added at a total enzyme activity of 100 U / g Astragalus membranaceus (500 U of cellulase, 300 U of xylanase, and 200 U of β-glucosidase per 1,000 U of total enzyme activity). The mixture was enzymatically hydrolyzed in a constant temperature water bath at 50°C for 3 hours with continuous stirring (200 rpm). The supernatant of the hydrolyzed Astragalus membranaceus was collected and concentrated under reduced pressure at 60°C (vacuum degree -0.09 MPa). The concentration was stopped when the relative density at 60°C reached 1.15, and the enzymatic hydrolysate of Astragalus membranaceus was obtained.

[0057] (3) Processing of other raw materials: 1) Sour jujube seed and longan pulp: 13 parts sour jujube seed and 10 parts longan pulp were dried at a low temperature of 45℃ and then pulverized and passed through a 200-mesh sieve; 2) Red dates, chicory, and kudzu root: 13 parts red dates, 8 parts chicory, and 4 parts kudzu root were dried at 65℃ and then pulverized through a 200-mesh sieve; 3) Lotus seeds and lily bulbs: Take 10 parts lotus seeds and 6 parts lily bulbs, mix them, add 10 times their weight of distilled water, and microwave extract twice (600W, 10 minutes / time). Combine the two extracts and concentrate them under reduced pressure at a vacuum degree of ≤-0.095 MPa. The endpoint is controlled at a relative density of 1.15 at 60℃ to obtain lotus seed and lily bulb concentrate.

[0058] (4) Extraction and concentration: Mix Angelica sinensis and Ganoderma lucidum fermentation product, Astragalus membranaceus enzymatic hydrolysate, Ziziphus jujuba seed powder, jujube powder, longan pulp powder, chicory powder and kudzu root powder, add 10 times the amount (about 2000 mL) of distilled water, reflux at 90℃ for 2 hours, filter, and collect the filtrate; mix the combined extract with lotus seed and lily bulb concentrate, and concentrate under reduced pressure at 60℃ to a relative density of 1.25.

[0059] (5) Refining into paste: Add 10% maltitol and 5% gum arabic as flavoring agent and stabilizer according to the weight of the paste, stir evenly, sterilize in water bath at 85°C for 15 minutes, fill into sterilized glass bottles while hot and seal.

[0060] The preparation methods of Examples 2-3 are similar to those of Example 1.

[0061] Preparation method of Comparative Example 1: (1) Ganoderma lucidum fermentation: 1) Preparation of Lactobacillus rhamnosus bacterial suspension: Take 100 mL of MRS liquid medium, sterilize at 121℃ for 15 minutes, and cool to 37℃; inoculate with 0.1 g of Lactobacillus rhamnosus CICC6137 lyophilized bacterial powder, and incubate at 37℃ for 24 hours to obtain Lactobacillus rhamnosus bacterial suspension with OD600≈1.0. Then inoculate it into MRS liquid medium at an inoculation rate of 2%, incubate at 37℃ until OD600≈1.5, and place it in a 4℃ refrigerator for later use.

[0062] 2) Inoculation: Ganoderma lucidum was pulverized and passed through an 80-mesh sieve, and inoculated with 3% Lactobacillus rhamnosus CICC6137 bacterial solution. Distilled water was added to adjust the moisture content to 45%.

[0063] 3) Fermentation: Solid-state fermentation at 37℃ for 72 hours, stirring once every 12 hours during the period, maintaining pH 6.0±0.3. After fermentation, the bacterial strain in the fermentation product is inactivated with 3% H2O2 solution, and then placed at 4℃ for use.

[0064] (2) Enzymatic hydrolysis of Astragalus membranaceus: Six portions of Astragalus membranaceus were pulverized and passed through a 60-mesh sieve. Phosphate buffer with pH 5.8 was added at a material-to-liquid ratio of 1:15. A compound enzyme was added at a total enzyme activity of 100 U / g Astragalus membranaceus (500 U of cellulase, 300 U of xylanase, and 200 U of β-glucosidase per 1,000 U of total enzyme activity). The mixture was enzymatically hydrolyzed in a constant temperature water bath at 50°C for 3 hours with continuous stirring (200 rpm). The supernatant of the hydrolyzed Astragalus membranaceus was collected and concentrated under reduced pressure at 60°C (vacuum degree -0.09 MPa). The concentration was stopped when the relative density reached 1.15 at 60°C, and the concentration was monitored using an online density meter.

[0065] (3) Processing of other raw materials: 1) Sour jujube seed, longan pulp, and angelica: 2 parts angelica, 13 parts sour jujube seed, and 10 parts longan pulp are dried at a low temperature of 45℃ and then pulverized through a 200-mesh sieve; 2) Red dates, chicory, and kudzu root: 13 parts red dates, 8 parts chicory, and 4 parts kudzu root were dried at 65℃ and then pulverized through a 200-mesh sieve; 3) Lotus seeds and lily bulbs: Take 10 parts lotus seeds and 6 parts lily bulbs, mix them, add 10 times their weight of distilled water, and microwave extract twice (600W, 10 minutes / time). Combine the two extracts and concentrate them under reduced pressure at a vacuum degree of ≤-0.095 MPa. The endpoint is controlled at a relative density of 1.15 at 60℃.

[0066] The remaining steps are the same as in Example 1.

[0067] Preparation method of Comparative Example 2: (1) Angelica sinensis fermentation: 1) Preparation of Lactobacillus rhamnosus bacterial suspension: Take 100 mL of MRS liquid medium, sterilize at 121℃ for 15 minutes, and cool to 37℃; inoculate with 0.1 g of Lactobacillus rhamnosus CICC6137 lyophilized bacterial powder, and incubate at 37℃ for 24 hours to obtain Lactobacillus rhamnosus bacterial suspension with OD600≈1.0. Then inoculate it into MRS liquid medium at an inoculation rate of 2%, incubate at 37℃ until OD600≈1.5, and place it in a 4℃ refrigerator for later use.

[0068] 2) Inoculation: Crush two portions of Angelica sinensis and pass them through an 80-mesh sieve. Inoculate with 3% Lactobacillus rhamnosus CICC6137 bacterial suspension and add distilled water to adjust the moisture content to 45%.

[0069] 3) Fermentation: Solid-state fermentation at 37℃ for 72 hours, stirring once every 12 hours during the period, maintaining pH 6.0±0.3. After fermentation, the bacterial strain in the fermentation product is inactivated with 3% H2O2 solution, and then placed at 4℃ for use.

[0070] (2) Enzymatic hydrolysis of Astragalus membranaceus: Six portions of Astragalus membranaceus were pulverized and passed through a 60-mesh sieve. Phosphate buffer at pH 5.8 was added at a material-to-liquid ratio of 1:15. A compound enzyme was added at a total enzyme activity of 100 U / g Astragalus membranaceus (500 U of cellulase, 300 U of xylanase, and 200 U of β-glucosidase per 1000 U of total enzyme activity). Enzymatic hydrolysis was performed in a 50℃ constant-temperature water bath for 3 hours with continuous stirring (200 rpm). The supernatant of the hydrolyzed Astragalus membranaceus was collected and concentrated under reduced pressure at 60℃ (vacuum degree -0.09 MPa). Concentration was stopped when the relative density reached 1.15 at 60℃, using an online density meter for monitoring.

[0071] (3) Processing of other raw materials: 1) Sour jujube seed, longan pulp, and angelica: 7 parts Ganoderma lucidum, 13 parts sour jujube seed, and 10 parts longan pulp were dried at a low temperature of 45℃ and then pulverized through a 200-mesh sieve; 2) Red dates, chicory, and kudzu root: 13 parts red dates, 8 parts chicory, and 4 parts kudzu root were dried at 65℃ and then pulverized through a 200-mesh sieve; 3) Lotus seeds and lily bulbs: Take 10 parts lotus seeds and 6 parts lily bulbs, mix them, add 10 times their weight of distilled water, and microwave extract twice (600W, 10 minutes / time). Combine the two extracts and concentrate them under reduced pressure at a vacuum degree of ≤-0.095 MPa. The endpoint is controlled at a relative density of 1.15 at 60℃.

[0072] The remaining steps are the same as in Example 1.

[0073] Preparation method of Comparative Example 3: (1) Enzymatic hydrolysis of Astragalus membranaceus: Six portions of Astragalus membranaceus were pulverized and passed through a 60-mesh sieve. Phosphate buffer at pH 5.8 was added at a material-to-liquid ratio of 1:15. A compound enzyme was added at a total enzyme activity of 100 U / g Astragalus membranaceus (500 U of cellulase, 300 U of xylanase, and 200 U of β-glucosidase per 1000 U of total enzyme activity). Enzymatic hydrolysis was performed in a 50℃ constant-temperature water bath for 3 hours with continuous stirring (200 rpm). The supernatant of the hydrolyzed Astragalus membranaceus was collected and concentrated under reduced pressure at 60℃ (vacuum degree -0.09 MPa). Concentration was stopped when the relative density reached 1.15 at 60℃, using an online density meter for monitoring.

[0074] (2) Processing of other raw materials: 1) Sour jujube seed, longan pulp, angelica, and ganoderma: 2 parts angelica, 7 parts ganoderma, 13 parts sour jujube seed and 10 parts longan pulp are dried at a low temperature of 45℃ and then pulverized through a 200-mesh sieve; 2) Red dates, chicory, and kudzu root: 13 parts red dates, 8 parts chicory, and 4 parts kudzu root were dried at 65℃ and then pulverized through a 200-mesh sieve; 3) Lotus seeds and lily bulbs: Take 10 parts lotus seeds and 6 parts lily bulbs, mix them, add 10 times their weight of distilled water, and microwave extract twice (600W, 10 minutes / time). Combine the two extracts and concentrate them under reduced pressure at a vacuum degree of ≤-0.095 MPa. The endpoint is controlled at a relative density of 1.15 at 60℃.

[0075] The remaining steps are the same as in Example 1.

[0076] Preparation method of Comparative Example 4: (1) Fermentation of Angelica sinensis and Ganoderma lucidum: 1) Preparation of Lactobacillus rhamnosus bacterial suspension: Take 100 mL of MRS liquid medium, sterilize at 121℃ for 15 minutes, and cool to 37℃; inoculate with 0.1 g of Lactobacillus rhamnosus ATCC9595 lyophilized bacterial powder, and incubate at 37℃ for 24 hours to obtain Lactobacillus rhamnosus bacterial suspension with OD600≈1.0. Then inoculate it into MRS liquid medium at an inoculation rate of 2%, incubate at 37℃ until OD600≈1.5, and place it in a 4℃ refrigerator for later use.

[0077] 2) Inoculation: Grind 2 parts of Angelica sinensis and 7 parts of Ganoderma lucidum into powder and pass them through an 80-mesh sieve. Mix them evenly and inoculate with 3% Lactobacillus rhamnosus ATCC9595 bacterial solution. Add distilled water to adjust the moisture content to 45%.

[0078] 3) Fermentation: Solid-state fermentation at 37℃ for 72 hours, stirring once every 12 hours during the period, maintaining pH 6.0±0.3. After fermentation, the bacterial strain in the fermentation product is inactivated with 3% H2O2 solution, and then placed at 4℃ for use.

[0079] The remaining steps are the same as in Example 1. Preparation method of Comparative Example 5: (1) Fermentation of Angelica sinensis and Ganoderma lucidum: 1) Preparation of Lactobacillus rhamnosus bacterial suspension: Take 100 mL of MRS liquid medium, sterilize at 121℃ for 15 minutes, and cool to 37℃; inoculate with 0.1 g of Lactobacillus rhamnosus CICC6137 lyophilized bacterial powder, and incubate at 37℃ for 24 hours to obtain Lactobacillus rhamnosus bacterial suspension with OD600≈1.0. Then inoculate it into MRS liquid medium at an inoculation rate of 2%, incubate at 37℃ until OD600≈1.5, and place it in a 4℃ refrigerator for later use.

[0080] 2) Inoculation: Grind 2 parts of Angelica sinensis and 7 parts of Ganoderma lucidum into powder and pass them through an 80-mesh sieve. Mix them evenly and inoculate with 3% Lactobacillus rhamnosus CICC6137 bacterial solution. Add distilled water to adjust the moisture content to 45%.

[0081] 3) Fermentation: Solid-state fermentation at 37℃ for 72 hours, stirring once every 12 hours during the period, maintaining pH 6.0±0.3. After fermentation, the bacterial strain in the fermentation product is inactivated with 3% H2O2 solution, and then placed at 4℃ for use.

[0082] (2) Processing of other raw materials: 1) Sour jujube seed and longan pulp: 13 parts sour jujube seed and 10 parts longan pulp were dried at a low temperature of 45℃ and then pulverized and passed through a 200-mesh sieve; 2) Astragalus, red dates, chicory, and kudzu root: 6 parts astragalus, 13 parts red dates, 8 parts chicory, and 4 parts kudzu root were dried at 65℃ and then pulverized through a 200-mesh sieve; 3) Lotus seeds and lily bulbs: Take 10 parts lotus seeds and 6 parts lily bulbs, mix them, add 10 times their weight of distilled water, and microwave extract twice (600W, 10 minutes / time). Combine the two extracts and concentrate them under reduced pressure at a vacuum degree of ≤-0.095 MPa. The endpoint is controlled at a relative density of 1.15 at 60℃.

[0083] The remaining steps are the same as in Example 1.

[0084] Efficacy Study of Food and Medicine Homologous Paste 1. Effects on the number of red blood cells in rats with blood deficiency syndrome Experimental animals: Male SD rats, purchased from Beijing Vital River Co., Ltd.

[0085] Mouse model of blood deficiency syndrome: The whole body was irradiated once with a 60Co-γ radiation source, with an irradiation dose of 3.5 Gy, a dose rate of 1.27 Gy / min, and an irradiation distance of 4 m.

[0086] Animal grouping: SD rats were divided into 16 groups of 8 rats each, namely control group, model group, Examples 1-3 groups, and Comparative Examples 1-11 groups. The prepared medicinal and edible extract was dissolved in distilled water at a ratio of 1:10. The dissolved extract was administered to the SD rats by gavage at a dose of 1 mL each time. The control group and model group were administered the same volume of distilled water by gavage. This gavage treatment continued for 2 weeks. After 2 weeks, the model group, Examples 1-3 groups, and Comparative Examples 1-9 groups were used to establish a mouse model of blood deficiency. Blood counts were performed on days 3, 7, 10, and 14 after modeling (50 μL of blood was collected from the tail vein each time). The red blood cell counts in each group are shown in Table 1. Table 1: Table 1 shows that the red blood cell count in the model group was significantly lower than that in the control group, proving the successful construction of the animal model. The red blood cell count in Example 1 was significantly higher than that in Examples 2-3 and Comparative Examples 1-11. The experimental results of Comparative Examples 1-4 and 9-10 indicate that co-fermentation of Ganoderma lucidum and Angelica sinensis can significantly increase the red blood cell count. The experimental results of Comparative Examples 5-8 and 11 indicate that enzymatic hydrolysis of Astragalus membranaceus by a specific complex enzyme can alleviate the reduction in red blood cell count caused by irradiation. In conclusion, the fermented products of Ganoderma lucidum and Angelica sinensis can synergistically improve the reduction in red blood cell count in mice with blood deficiency syndrome in mice with Astragalus membranaceus enzymatic hydrolysis.

[0087] 2. Effects on mouse sleep Laboratory animals: Male Kunming mice, purchased from Beijing Vital River Co., Ltd. Animal grouping: Mice were divided into 16 groups of 10 each, namely control group, positive group, Example 1-3 groups, and Comparative Example 1-11 groups. The prepared medicinal and edible homology paste was dissolved in distilled water at a ratio of 1:10. The medicinal and edible homology paste dissolved in water was administered to Kunming mice by gavage. The control group was administered the same volume of distilled water by gavage, and the positive group was administered the same volume of diazepam (1 mg / mL) by gavage. Each administration was 200 uL and continued for 2 weeks.

[0088] Direct sleep experiment: Each group of animals was administered the corresponding test substance daily. After the last administration, mice were observed for sleep patterns within 60 minutes post-gavage. Sleep was defined as the absence of the righting reflex for more than 60 seconds, and awakening was defined as the return of the righting reflex. The number of animals falling asleep and their respective sleep durations were recorded for each group. The results showed that no direct sleep behavior was observed in any group of mice. Pentobarbital sodium sleep duration prolongation experiment: Thirty minutes after the last administration to mice, sodium pentobarbital was injected intraperitoneally at a dose of 50 mg / kg (based on mouse body weight, mice fell asleep in 100% of cases but the sleep time was short). The sleep duration (time from the disappearance of the righting reflex to the reappearance of the righting reflex) of each group of mice was observed and recorded.

[0089] Sodium barbital sleep latency experiment: Thirty minutes after the last administration, sodium barbital was injected intraperitoneally at a dose of 230 mg / kg (based on mouse body weight, ensuring 100% sleep in mice). The time between intraperitoneal injection and the disappearance of the righting reflex was recorded as the sleep latency, with the disappearance of the righting reflex 60 seconds as the indicator.

[0090] Table 2: Table 2 shows that the sleep quality of Example 1 was significantly better than that of Examples 2-3 and Comparative Examples 1-11. Direct sleep experiments indicate that this invention does not directly induce sleep, but rather works synergistically with sodium pentobarbital by regulating the sleep threshold. Comparative Examples 1-4 and 9-10 show that Ganoderma lucidum and Angelica sinensis ferments can improve sleep quality in mice; Comparative Examples 5-8 and 11 show that Astragalus membranaceus requires specific enzymatic hydrolysis to achieve a better effect on improving sleep quality. In conclusion, the results of Examples 1-3 and Comparative Examples 1-11 indicate that the synergistic effect of Ganoderma lucidum and Angelica sinensis ferments with Astragalus membranaceus hydrolysate on improving sleep quality is the best.

[0091] 3. Antioxidant experiment (DPPH free radical scavenging experiment): For Examples 1-3 and Comparative Examples 1-11, the prepared medicinal and edible extract was dissolved in distilled water at a ratio of 1:10. After dissolution, 3 ml of the solution was added to 3 ml of DPPH anhydrous ethanol solution (0.1 mmol / L), and the mixture was shaken well and incubated at room temperature in the dark for 30 min. The absorbance was measured at 517 nm (A1). Similarly, 3 ml of anhydrous ethanol solution was added to 3 ml of the sample to be tested, and the mixture was shaken well and incubated at room temperature in the dark for 30 min. The absorbance was measured at 517 nm (A2). Finally, 3 ml of distilled water was added to 3 ml of DPPH anhydrous ethanol solution (0.1 mmol / L), and the mixture was shaken well and incubated at room temperature in the dark for 30 min. The absorbance was measured at 517 nm (A0). The DPPH free radical scavenging rate was calculated using the following formula, as shown in Table 3.

[0092] DPPH radical scavenging rate = (1 - (A1 - A2) / A0) × 100% Table 3:

[0093] Table 3 shows that the DPPH free radical scavenging rate of Examples 1-3 was much higher than that of Comparative Examples 1-11. The experimental results of Comparative Examples 1-4 and 9-10 demonstrate that Ganoderma lucidum and Angelica sinensis, after fermentation with Lactobacillus rhamnosus CICC6137, exhibit synergistic antioxidant effects. The experimental results of Comparative Examples 5-8 and 11 indicate that Astragalus membranaceus requires enzymatic hydrolysis by a specific complex enzyme to exert its superior antioxidant function. In conclusion, the synergistic effect of Ganoderma lucidum and Angelica sinensis fermentation products with Astragalus membranaceus enzymatic hydrolysate can achieve the best antioxidant effect.

[0094] 4. Analgesia experiment: Experimental animals: Kunming mice, 18-22g, purchased from Beijing Vital River Co., Ltd.

[0095] Experimental groups: A total of 12 groups were set up, including a control group, a positive group, Examples 1-3, Comparative Examples 1-4, Comparative Examples 9-10, and Comparative Example 12, with 10 mice in each group. The prepared medicinal and edible homology paste for each group was dissolved in distilled water at a ratio of 1:10. The dissolved medicinal and edible homology paste was administered to the mice by gavage at a dose of 200 uL each time. The control group was administered the same volume of distilled water by gavage, and the positive group was administered the same volume of aspirin at a dose of 400 mg / kg by gavage. The gavage was continued for 2 weeks.

[0096] Experimental methods: Mice were fasted for 12 hours before the experiment. One hour after the last administration on the day of the experiment, each mouse was injected intraperitoneally with 200 μL of 0.8% acetic acid saline solution. The number of writhing movements of the mice within 15 minutes was observed, and the writhing inhibition rate was calculated as shown in Table 4.

[0097] The writhing inhibition rate (%) = [(control group - treatment group) / control group] × 100%, where the treatment group includes the positive group, Example 1-3 groups, Comparative Example 1-4 groups, Comparative Example 9-10 groups and Comparative Example 12 groups.

[0098] Table 4:

[0099] As shown in Table 4, the number of writhing movements in Example 1 was significantly lower than that in Examples 2-3, Comparative Examples 1-4, and Comparative Examples 9, 10, and 12. Compared to Example 1, the experimental results of Comparative Examples 1-4 and 9-10 indicate that fermentation with *Lactobacillus rhamnosus* CICC6137 can reduce the number of writhing movements in Angelica sinensis and Ganoderma lucidum. The results of Comparative Example 12 indicate that, compared to Example 1, Lilium brownii has an auxiliary analgesic effect.

[0100] During the research and development of this invention, extensive strain screening (such as comparing CICC6137, ATCC9595, and DSM20021) and enzyme combination optimization were conducted. Ultimately, the optimal combination of CICC6137 and a specific complex enzyme ratio was determined. This process involved dozens of iterative experiments. Results showed that CICC6137 increased polysaccharide yield by 40% after fermentation (superior to 20-25% for other strains), and the yield of astragalus flavonoids from the complex enzyme hydrolysis reached 90 mg / g (higher than the 60-70 mg / g of a single enzyme), leading to unexpected... Synergistic synergistic effects: In anemic rat models, red blood cell count increased by 30% (Table 1, 15-20% higher than the control group), sleep was prolonged by 25% (Table 2), DPPH clearance rate was 95% (Table 3), and writhing inhibition rate was 36.5% (Table 4). These effects far exceeded the expected levels of conventional fermentation or enzymatic hydrolysis in existing technologies (literature reports usually show an improvement of 10-20%). This process combination triggered an unexpected overall synergistic effect (such as a 30% increase in red blood cells and a 25% prolongation of sleep), demonstrating the originality and technological contribution of this specific process.

[0101] The embodiments described above are merely illustrative of specific implementations of the present invention, and while the descriptions are detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention.

Claims

1. A composite starter culture, characterized in that, The complex fermentation agent comprises Lactobacillus rhamnosus CICC6137, is used for fermenting a mixture of Ganoderma lucidum and Angelica sinensis, and has a bacteria liquid OD600 of about 1.5, an inoculation amount of 3-5% (v / w), and a fermentation condition of 37°C solid-state fermentation for 48-72h and pH 5.0-6.

5.

2. A Shensan Sanpao with the effects of nourishing blood, soothing the nerves, improving sleep and resisting oxidative damage, characterized in that, The preparation is composed of the following raw materials in parts by weight: 10-16 parts of Chinese date kernel, 10-16 parts of red dates, 8-12 parts of longan arillus, 8-12 parts of lotus seeds, 1-3 parts of angelica, 5-10 parts of ganoderma, 5-8 parts of astragalus, 6-10 parts of chicory, 5-8 parts of lily, and 3-5 parts of kudzu root, wherein the ganoderma and the angelica are fermented by the complex fermentation agent of claim 1.

3. The paste of claim 2, wherein The preparation is composed of the following raw materials in parts by weight: 13 parts of Chinese date kernel, 13 parts of red dates, 10 parts of longan arillus, 10 parts of lotus seeds, 2 parts of angelica, 7 parts of ganoderma, 6 parts of astragalus, 8 parts of chicory, 6 parts of lily, and 4 parts of kudzu root.

4. The paste of claim 2, wherein The ganoderma and the angelica are subjected to fermentation treatment by Lactobacillus rhamnosus to improve the bioavailability of active ingredients.

5. The paste of claim 2, wherein The astragalus is subjected to enzymatic hydrolysis treatment by a complex enzyme, and the complex enzyme is composed of cellulase, xylanase, and β-glucosidase.

6. A method of preparing the functional food paste as claimed in any one of claims 2 to 5, wherein, The preparation comprises the following steps: S1, uniformly mixing 5-10 parts of ganoderma and 1-3 parts of angelica after being respectively crushed, and adding Lactobacillus rhamnosus for fermentation treatment to obtain angelica-ganoderma fermentation product; S2, crushing 5-8 parts of astragalus, and subjecting to enzymatic hydrolysis treatment by a complex enzyme to obtain astragalus enzymatic hydrolysate, wherein in 1,000 U of total enzyme activity of the complex enzyme, 500 U is cellulase, 300 U is xylanase, and 200 U is β-glucosidase; S3, mixing 8-12 parts of lotus seeds and 5-8 parts of lily, adding water, and performing microwave extraction to obtain lotus-lily concentrated liquid after concentration; S4, crushing 10-16 parts of Chinese date kernel, 10-16 parts of red dates, 8-12 parts of longan arillus, 6-10 parts of chicory, and 3-5 parts of kudzu root, respectively, mixing the angelica-ganoderma fermentation product obtained in step S1 and the astragalus enzymatic hydrolysate treated in step S2, and performing water extraction and filtration to obtain filtrate; and mixing the filtrate and the lotus-lily concentrated liquid obtained in step S3, and concentrating into paste.

7. The method of claim 6, wherein, In step S1, the inoculation amount of Lactobacillus rhamnosus is 3-5%, the fermentation temperature is 37°C, and the fermentation time is 48-72 hours.

8. The method of claim 7, wherein, In step S1, Lactobacillus rhamnosus CICC6137 is selected.

9. The method of claim 8, wherein, In step S1, the OD600 of Lactobacillus rhamnosus is about 1.

5. In step S2, the enzymatic hydrolysis treatment by the complex enzyme is performed under the following conditions: enzymatic hydrolysis temperature of 50-55°C, pH value of 5.0-6.0, and enzymatic hydrolysis time of 2-4 hours.

10. Use of the preparation of any one of claims 2-5 in the preparation of a product with the functions of nourishing blood, soothing nerves, improving sleep, resisting oxidative damage, and relieving pain.

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