Dendrobium officinale fermentation liquor composition with anti-inflammatory effect and preparation method thereof

Abstract

The invention belongs to the field of health food, and particularly relates to a dendrobium officinale fermentation liquor composition with an anti-inflammatory effect and a preparation method thereof.The composition is prepared from 30-40 parts of dendrobium officinale fermentation liquor, 15-25 parts of cyclocarya paliurus juice, 10-20 parts of kiwi fruit juice, 8-12 parts of sugarcane juice, 3-5 parts of polydextrose, 2-4 parts of lemon juice and 0.5-1 part of mint. The invention further optimizes a process method for fermenting the dendrobium officinale. According to the dendrobium officinale fermentation liquor composition, the anti-inflammatory effect is remarkably improved, the immunocompetence in a mouse body can be adjusted, the remarkable protection effect on the intestinal tract of the mouse is achieved, and the total phenol content and the total sugar content of the dendrobium officinale fermentation liquor can be remarkably improved through the dendrobium officinale fermentation process.

Description

Technical Field

[0001] This invention relates to the field of food technology, and in particular to a Dendrobium officinale fermentation liquid composition with anti-inflammatory effects. Background Technology

[0002] Dendrobium officinale, commonly known as black-jointed grass, is mainly distributed in Anhui, Zhejiang, and Fujian provinces of China. Its scientific name was officially established in the *Flora of China* in 1999 and included in the *Chinese Pharmacopoeia* in 2005. Studies have shown that Dendrobium officinale contains polysaccharides, polyphenols, flavonoids, alkaloids, and other components, possessing various active functions such as lowering blood sugar, anti-inflammation, anti-oxidation, and enhancing immunity. As a plant of the *Dendrobium* genus used for both food and medicine, Dendrobium officinale can be consumed directly or used to develop various products, such as Dendrobium officinale wine, beverages, and biscuits. Furthermore, it can be developed into health products or traditional Chinese medicine preparations, such as capsules, granules, and oral liquids. However, patent document CN118489818A mentions that a fermented Dendrobium officinale sparkling water suffers from insufficient anti-inflammatory effects and difficulty in the effective utilization of its components.

[0003] Anti-inflammatory effects refer to helping the body cope with inflammatory processes caused by infection, injury, or immune abnormalities by inhibiting inflammatory responses, reducing tissue damage, or alleviating symptoms. The core lies in regulating the release of inflammatory mediators and balancing the immune response. The mechanisms of anti-inflammatory action are divided into two types: inhibiting inflammatory mediators and regulating immune cell activity. Inhibiting inflammatory mediators involves anti-inflammatory substances reducing their production by blocking related enzymes or interfering with signaling pathways; regulating immune cell activity involves restoring immune balance through cell migration and reducing cytokine secretion. Patent document CN118725154A mentions a low molecular weight Dendrobium officinale polysaccharide with both antioxidant and anti-inflammatory effects, but the product has insufficient anti-inflammatory efficacy. Patent document CN115572738B mentions a Dendrobium officinale fermented liquid with anti-inflammatory, soothing, and repairing effects, but it targets skin repair and cannot demonstrate its anti-inflammatory effects in vivo.

[0004] Therefore, it is very important to study a composition based on Dendrobium officinale fermentation liquid that enhances anti-inflammatory effects and has a rich flavor. Summary of the Invention

[0005] One objective of this invention is to overcome the aforementioned technical shortcomings and provide a Dendrobium officinale fermentation liquid composition with anti-inflammatory effects. The composition is mainly composed of Dendrobium officinale fermentation liquid, Eucommia ulmoides juice, kiwi juice, and sugarcane juice. It has multi-target anti-inflammatory and antioxidant effects. The active ingredients from different sources and with different molecular structures can act on multiple links in the inflammatory and oxidative stress pathways, forming a more comprehensive protective network.

[0006] Another objective of this invention is to improve the utilization of active ingredients. The organic acids in the added kiwi juice and lemon juice can regulate the dissolution and absorption of active ingredients, and the vitamins in kiwi can also reduce the oxidative loss of other antioxidants during processing and in the body.

[0007] Another objective of this invention is to obtain a composition that can comprehensively regulate physical health. The addition of polydextrose, lemon juice, and mint enhances the composition's regulatory effect on the body.

[0008] Another objective of this invention is to provide a method for preparing Dendrobium officinale fermentation liquid. By limiting the fermentation bacteria and optimizing the reaction process, a Dendrobium officinale fermentation liquid with better fermentation effect can be obtained.

[0009] Another objective of this invention is to provide a method for preparing a Dendrobium officinale fermentation liquid composition with anti-inflammatory effects, which is simple and effective.

[0010] To achieve the above objectives, this invention discloses a Dendrobium officinale fermentation liquid composition with anti-inflammatory effects, comprising the following raw materials by weight: 30-40 parts Dendrobium officinale fermentation liquid, 15-25 parts Eucommia ulmoides juice, 10-20 parts kiwifruit juice, and 8-12 parts sugarcane juice. The Dendrobium officinale fermentation liquid contains abundant Dendrobium officinale oligosaccharides, polysaccharides, polyphenols, and flavonoids, which are core components for regulating intestinal flora, increasing short-chain fatty acid production, inhibiting the NF-κB inflammatory pathway, and reducing TNF-α and IL-6 pro-inflammatory factors. The saponins and flavonoids in Eucommia ulmoides juice have anti-inflammatory effects, regulate immune cell function, and also possess antioxidant properties. The kiwifruit juice contains abundant vitamin C. Polyphenols and kiwifruit protease have a strong antioxidant effect, scavenging free radicals; sugarcane juice contains polyphenols and flavonoids, which work synergistically with kiwifruit juice for antioxidant effects; the above components work together to create a multi-target, multi-level synergistic effect: kiwifruit juice and sugarcane juice form an antioxidant network, reducing free radicals and oxidative stress that cause tissue damage and chronic inflammation from the source; Dendrobium officinale fermentation liquid is the core, inhibiting inflammatory channels to reduce excessive immune response, optimizing intestinal flora structure, and promoting the production of anti-inflammatory short-chain fatty acids, achieving systemic anti-inflammatory effects from the gut-immune axis; and Eucommia ulmoides juice has the ability to rapidly inhibit acute inflammatory responses, forming a complete anti-inflammatory system from prevention, regulation to intervention.

[0011] Preferably, the Dendrobium officinale fermentation liquid is derived from Dendrobium officinale juice through fermentation, and the flavonoid content of the Eucommia ulmoides juice is 3-3.5%.

[0012] Preferably, the fermentation bacteria in the Dendrobium officinale juice fermentation is a compound bacterial agent of Lactobacillus acidophilus and Lactobacillus plantarum. Fermenting Dendrobium officinale juice with Lactobacillus acidophilus and Lactobacillus plantarum can significantly increase the content of polyphenols and flavonoids, enhance anti-inflammatory effects, and degrade large-molecule polysaccharides into small-molecule polysaccharides, making them easier for the human body to absorb.

[0013] Preferably, the ratio of Lactobacillus acidophilus to Lactobacillus plantarum is 0.6-3:1, and the inoculation amount is 1-9% of Dendrobium officinale juice.

[0014] As a preferred option, the following components are also included, by weight: 3-5 parts polydextrose, 2-4 parts lemon juice, 0.5-1 part peppermint, and purified water to make up to 120 parts. Lemon juice is rich in vitamin C, citric acid, and flavonoids, which work synergistically with kiwi juice and sugarcane juice to build a powerful antioxidant network. Polydextrose, along with oligosaccharides and polysaccharides in Dendrobium officinale, acts as prebiotics to optimize the intestinal flora structure, while peppermint can help Eucommia ulmoides juice quickly suppress acute inflammation.

[0015] Preferably, the particle size of nano zinc oxide is required to be 30-60 nm.

[0016] This invention also provides a method for preparing Dendrobium officinale fermentation broth, comprising the following steps: S1. After washing the Dendrobium officinale stems, juice them using a high-speed blender at a material-to-liquid ratio of 1:20 (m / v), filter the juice, and then sterilize it for later use. S2. Cultivate the compound bacteria, and after three generations, wash three times with physiological saline for later use. S3. After inoculating the activated compound bacteria into the sterilized Dendrobium officinale juice for fermentation, Dendrobium officinale fermentation liquid is obtained.

[0017] Preferably, the sterilization process in step S1 involves sterilizing the bacteria at 121°C for 10 minutes in an autoclave; the culture in step S2 involves incubating the bacteria in MRS broth medium at 37°C for 48 hours, subculturing twice, activating the bacteria, and then culturing to the third generation to obtain an initial bacterial concentration of 2.6 × 10⁻⁶. 8 CFU / mL; the fermentation conditions for S3 are: fermentation temperature of 31-43℃ and fermentation time of 12-60h.

[0018] This invention also provides a method for preparing Dendrobium officinale fermentation broth, comprising the following steps: S1. Add the Dendrobium officinale fermentation liquid, Eucommia ulmoides juice, kiwi juice, and sugarcane juice to the mixing tank in proportion, stir, add polydextrose and continue stirring, then add lemon juice and mint pieces, and stir lightly to obtain a mixture. S2. The mixture is homogenized using a homogenizer, and then water is added to make up to 120 parts to obtain a preliminary Dendrobium officinale fermentation liquid composition. S3. The preliminary Dendrobium officinale fermentation liquid composition is sterilized to obtain a Dendrobium officinale fermentation liquid composition with anti-inflammatory effects.

[0019] Preferably, the kiwi juice and sugarcane juice in step S1 are fresh kiwi and sugarcane juices that have been washed, peeled, pressed, coarsely filtered, and sterilized. The sterilization temperature is 95℃ and the sterilization time is 30s. The stirring in step S1 is 400-600rpm and stirred for 10-20min.

[0020] Preferably, the pressure of the homogenizer in step S2 is 15-25 MPa, and the sterilization treatment in step S3 is 400-600 MPa for 5-10 minutes.

[0021] The beneficial effects of this invention are as follows: The Dendrobium officinale fermentation liquid composition of this application has multi-target synergistic anti-inflammatory effects. Different components in the composition exert synergistic effects through complementary mechanisms. By scientifically compounding Dendrobium officinale fermentation liquid, Eucommia ulmoides juice, kiwi juice and sugarcane juice, multi-target, deep anti-inflammatory effects from the intestinal root are achieved, and the synergistic enhancement of antioxidant and metabolic regulation is combined. At the same time, the addition of lemon juice and mint solves the palatability problem that is common in functional beverages.

[0022] The Dendrobium officinale fermentation liquid composition of this invention has the effect of assisting in regulating metabolism. In addition to assisting in anti-inflammatory effects, Eucommia ulmoides juice also has the effect of assisting in lowering blood sugar and blood lipids. Kiwi juice has the effect of assisting digestion. The dietary fiber and organic acids in sugarcane juice can work synergistically with kiwi juice to promote digestion.

[0023] The Dendrobium officinale fermentation liquid composition of the present invention regulates immunity and has antioxidant properties. The polysaccharides and flavonoids in the Dendrobium officinale fermentation liquid can activate immune cells and enhance the body's disease resistance. They work synergistically with Eucommia ulmoides juice to fight oxidation and improve immunity. Kiwi juice is rich in vitamin C, which synergistically promotes white blood cell activity. It also contains vitamin E and polyphenols to assist in antioxidant activity.

[0024] The present invention relates to a method for preparing Dendrobium officinale fermentation liquid, which uses Lactobacillus acidophilus and Lactobacillus plantarum to ferment Dendrobium officinale juice, thereby increasing the content of active substances and significantly improving the content of anti-inflammatory and antioxidant components such as total phenols and total flavonoids; optimizing absorption and utilization by degrading large-molecule polysaccharides into small molecules, thereby improving bioavailability.

[0025] The present invention discloses a method for preparing a Dendrobium officinale fermentation liquid composition, which can produce a Dendrobium officinale fermentation liquid composition with anti-inflammatory effects, and the process is simple. Detailed Implementation

[0026] The present invention will be further described in detail below with reference to specific embodiments.

[0027] General Example: Dendrobium officinale fermentation broth composition: 30-40 parts Dendrobium officinale fermentation broth, 15-25 parts Eucommia ulmoides juice, 10-20 parts kiwi juice, 8-12 parts sugarcane juice, 3-5 parts polydextrose, 2-4 parts lemon juice, 0.5-1 part peppermint, and purified water to a total of 120 parts; A method for preparing Dendrobium officinale fermentation broth: S1. Take the washed Dendrobium officinale stems and juice them using a blender at a material-to-liquid ratio of 1:20 (m / v). Filter the juice and then sterilize it in an autoclave at 121℃ for 10 minutes.

[0028] S2. Inoculate *Lactobacillus plantarum* and *Lactobacillus acidophilus* at a ratio of (0.3-3):1 into MRS broth medium, incubate at 37℃ for 48 hours, then perform a second subculture. After activation, culture to the third generation, wash three times with physiological saline, with an initial bacterial concentration of 2.6 × 10⁻⁶. 8 CFU / mL; S3. Inoculate the activated Lactobacillus plantarum and Lactobacillus acidophilus into the Dendrobium officinale juice prepared in S1. The inoculation amount is 1-9% of the Dendrobium officinale juice. The fermentation temperature is 31-43℃ and the fermentation time is 24-60h. The plant lactobacillus and lactobacillus acidophilus used were obtained from the Henan Provincial Industrial Microbial Strain Engineering Technology Research Center, and the Dendrobium officinale stems were obtained from Zhejiang Tiefengtang Health Technology Co., Ltd.

[0029] A method for preparing a Dendrobium officinale fermentation broth composition: S1. Add the Dendrobium officinale fermentation liquid, Eucommia ulmoides juice, kiwi juice, and sugarcane juice to the mixing tank in proportion. After stirring, add polydextrose and continue stirring. Then add lemon juice and mint pieces and stir gently to obtain a mixture. The stirring speed is 400-600 rpm and the stirring time is 10-20 minutes. S2. The mixture is homogenized using a homogenizer, and then water is added to make up to 120 parts. The pressure of the homogenizer is 15-20 MPa to obtain a preliminary Dendrobium officinale fermentation liquid composition. S3. The initial Dendrobium officinale fermentation broth composition is sterilized at 400-600 MPa for 5-10 minutes to obtain a Dendrobium officinale fermentation broth composition with anti-inflammatory effects. Example 1:

[0030] Composition of Dendrobium officinale fermentation liquid: 35 parts Dendrobium officinale fermentation liquid, 20 parts Eucommia ulmoides juice, 15 parts kiwi juice, 10 parts sugarcane juice, 4 parts polydextrose, 3 parts lemon juice, 0.8 parts mint, and purified water to make up to 120 parts. A method for preparing Dendrobium officinale fermentation broth: S1. Take the washed Dendrobium officinale stems and juice them using a blender at a material-to-liquid ratio of 1:20 (m / v). Filter the juice and then sterilize it in an autoclave at 121℃ for 10 minutes. S2. Inoculate *Lactobacillus plantarum* and *Lactobacillus acidophilus* at a 1:1 ratio into MRS broth medium, incubate at 37°C for 48 hours, then perform a second subculture. After activation, culture to the third generation, wash three times with physiological saline, with an initial bacterial concentration of 2.6 × 10⁻⁶. 8 CFU / mL; S3. Inoculate the activated Lactobacillus plantarum and Lactobacillus acidophilus into the Dendrobium officinale juice prepared in S1. The inoculation amount is 5% of the Dendrobium officinale juice. The fermentation temperature is 37℃ and the fermentation time is 36h. A method for preparing a Dendrobium officinale fermentation broth composition: S1. Add the Dendrobium officinale fermentation liquid, Eucommia ulmoides juice, kiwi juice, and sugarcane juice to the mixing tank in proportion, stir, add polydextrose and continue stirring, then add lemon juice and mint pieces, and stir gently to obtain a mixture; the stirring speed is 500 rpm and the stirring time is 15 min. S2. The mixture is homogenized using a homogenizer, and then water is added to make up to 120 parts. The pressure of the homogenizer is 18 MPa to obtain a preliminary Dendrobium officinale fermentation liquid composition. S3. The initial Dendrobium officinale fermentation broth composition is sterilized at 500 MPa for 8 minutes to obtain a Dendrobium officinale fermentation broth composition with anti-inflammatory effects.

[0031] Unless otherwise specified, the preparation methods in the following embodiments are the same as those in Example 1. Example 2:

[0032] The composition of Dendrobium officinale fermentation liquid is as follows: 30 parts Dendrobium officinale fermentation liquid, 20 parts Eucommia ulmoides juice, 15 parts kiwi juice, 10 parts sugarcane juice, 4 parts polydextrose, 3 parts lemon juice, 0.8 parts mint, and purified water to make up to 120 parts. Example 3:

[0033] The composition of Dendrobium officinale fermentation liquid is as follows: 40 parts Dendrobium officinale fermentation liquid, 20 parts Eucommia ulmoides juice, 15 parts kiwi juice, 10 parts sugarcane juice, 4 parts polydextrose, 3 parts lemon juice, 0.8 parts mint, and purified water to make up to 120 parts. Example 4:

[0034] The composition of Dendrobium officinale fermentation liquid is as follows: 35 parts Dendrobium officinale fermentation liquid, 15 parts Eucommia ulmoides juice, 15 parts kiwi juice, 10 parts sugarcane juice, 4 parts polydextrose, 3 parts lemon juice, 0.8 parts mint, and purified water to make up to 120 parts. Example 5:

[0035] The composition of Dendrobium officinale fermentation liquid is as follows: 35 parts Dendrobium officinale fermentation liquid, 25 parts Eucommia ulmoides juice, 15 parts kiwi juice, 10 parts sugarcane juice, 4 parts polydextrose, 3 parts lemon juice, 0.8 parts mint, and purified water to make up to 120 parts. Example 6:

[0036] The composition of Dendrobium officinale fermentation liquid is as follows: 35 parts Dendrobium officinale fermentation liquid, 20 parts Eucommia ulmoides juice, 10 parts kiwi juice, 10 parts sugarcane juice, 4 parts polydextrose, 3 parts lemon juice, 0.8 parts peppermint, and purified water to make up to 120 parts. Example 7:

[0037] The composition of Dendrobium officinale fermentation liquid is as follows: 35 parts Dendrobium officinale fermentation liquid, 20 parts Eucommia ulmoides juice, 20 parts kiwi juice, 10 parts sugarcane juice, 4 parts polydextrose, 3 parts lemon juice, 0.8 parts peppermint, and purified water to make up to 120 parts. Example 8:

[0038] Composition of Dendrobium officinale fermentation liquid: 35 parts Dendrobium officinale fermentation liquid, 20 parts Eucommia ulmoides juice, 15 parts kiwi juice, 8 parts sugarcane juice, 4 parts polydextrose, 3 parts lemon juice, 0.8 parts mint, and purified water to make up to 120 parts. Example 9:

[0039] Composition of Dendrobium officinale fermentation liquid: 35 parts Dendrobium officinale fermentation liquid, 20 parts Eucommia ulmoides juice, 15 parts kiwi juice, 12 parts sugarcane juice, 4 parts polydextrose, 3 parts lemon juice, 0.8 parts mint, and purified water to make up to 120 parts. Example 10:

[0040] The composition of Dendrobium officinale fermentation liquid is as follows: 35 parts Dendrobium officinale fermentation liquid, 20 parts Eucommia ulmoides juice, 15 parts kiwi juice, 10 parts sugarcane juice, 3 parts polydextrose, 3 parts lemon juice, 0.8 parts mint, and purified water to make up to 120 parts. Example 11:

[0041] The composition of Dendrobium officinale fermentation liquid is as follows: 35 parts Dendrobium officinale fermentation liquid, 20 parts Eucommia ulmoides juice, 15 parts kiwi juice, 10 parts sugarcane juice, 5 parts polydextrose, 3 parts lemon juice, 0.8 parts mint, and purified water to make up to 120 parts. Example 12:

[0042] The composition of Dendrobium officinale fermentation liquid is as follows: 35 parts Dendrobium officinale fermentation liquid, 20 parts Eucommia ulmoides juice, 15 parts kiwi juice, 10 parts sugarcane juice, 4 parts polydextrose, 2 parts lemon juice, 0.8 parts mint, and purified water to make up to 120 parts. Example 13:

[0043] The composition of Dendrobium officinale fermentation liquid is as follows: 35 parts Dendrobium officinale fermentation liquid, 20 parts Eucommia ulmoides juice, 15 parts kiwi juice, 10 parts sugarcane juice, 4 parts polydextrose, 4 parts lemon juice, 0.8 parts mint, and purified water to make up to 120 parts. Example 14:

[0044] Composition of Dendrobium officinale fermentation liquid: 35 parts Dendrobium officinale fermentation liquid, 20 parts Eucommia ulmoides juice, 15 parts kiwi juice, 10 parts sugarcane juice, 4 parts polydextrose, 3 parts lemon juice, 0.5 parts mint, and purified water to make up to 120 parts. Example 15:

[0045] The composition of Dendrobium officinale fermentation liquid is as follows: 35 parts Dendrobium officinale fermentation liquid, 20 parts Eucommia ulmoides juice, 15 parts kiwi juice, 10 parts sugarcane juice, 4 parts polydextrose, 3 parts lemon juice, 1 part mint, and purified water to make up to 120 parts.

[0046] The following examples investigate the technical parameters in the preparation of Dendrobium officinale fermentation broth by varying the proportion of inoculum, inoculum size, fermentation temperature, and fermentation time. The composition formulation of Example 1 is used as a baseline. Example 16:

[0047] A method for preparing Dendrobium officinale fermentation broth: S1, Same as in Example 1; S2, the inoculation ratio of Lactobacillus plantarum and Lactobacillus acidophilus is 3:1, and the rest is the same as in Example 1; S3, Same as Example 1. Example 17:

[0048] A method for preparing Dendrobium officinale fermentation broth: S1, Same as in Example 1; S2, the inoculation ratio of Lactobacillus plantarum and Lactobacillus acidophilus is 2:1, and the rest is the same as in Example 1; S3, Same as Example 1. Example 18:

[0049] A method for preparing Dendrobium officinale fermentation broth: S1, Same as in Example 1; S2, the inoculation ratio of Lactobacillus plantarum and Lactobacillus acidophilus is 0.5:1, and the rest is the same as in Example 1; S3, Same as Example 1. Example 19:

[0050] A method for preparing Dendrobium officinale fermentation broth: S1, Same as in Example 1; S2, the inoculation ratio of Lactobacillus plantarum and Lactobacillus acidophilus is 0.3:1, and the rest is the same as in Example 1; S3, Same as Example 1. Example 20:

[0051] A method for preparing Dendrobium officinale fermentation broth: S1, Same as in Example 1; S2, Same as in Example 1; S3, the inoculation amount is 1% of Dendrobium officinale juice, and the other aspects are the same as in Example 1. Example 21:

[0052] A method for preparing Dendrobium officinale fermentation broth: S1, Same as in Example 1; S2, Same as in Example 1; S3, the inoculation amount is 3% of Dendrobium officinale juice, and the other aspects are the same as in Example 1. Example 22:

[0053] A method for preparing Dendrobium officinale fermentation broth: S1, Same as in Example 1; S2, Same as in Example 1; S3, the inoculation amount is 7% of Dendrobium officinale juice, and the other aspects are the same as in Example 1. Example 23:

[0054] A method for preparing Dendrobium officinale fermentation broth: S1, Same as in Example 1; S2, Same as in Example 1; S3, the inoculation amount is 9% of Dendrobium officinale juice, and the other aspects are the same as in Example 1. Example 24:

[0055] A method for preparing Dendrobium officinale fermentation broth: S1, Same as in Example 1; S2, Same as in Example 1; S3, the fermentation temperature is 31℃, and the other conditions are the same as in Example 1. Example 25:

[0056] A method for preparing Dendrobium officinale fermentation broth: S1, Same as in Example 1; S2, Same as in Example 1; S3, the fermentation temperature is 34℃, and the other conditions are the same as in Example 1. Example 26:

[0057] A method for preparing Dendrobium officinale fermentation broth: S1, Same as in Example 1; S2, Same as in Example 1; S3, the fermentation temperature is 40℃, and the other conditions are the same as in Example 1. Example 27:

[0058] A method for preparing Dendrobium officinale fermentation broth: S1, Same as in Example 1; S2, Same as in Example 1; S3, fermentation temperature is 43℃, the other conditions are the same as in Example 1. Example 28:

[0059] A method for preparing Dendrobium officinale fermentation broth: S1, Same as in Example 1; S2, Same as in Example 1; S3, fermentation time 24h, other aspects are the same as in Example 1. Example 29:

[0060] A method for preparing Dendrobium officinale fermentation broth: S1, Same as in Example 1; S2, Same as in Example 1; S3, fermentation time 48h, other aspects are the same as in Example 1. Example 30:

[0061] A method for preparing Dendrobium officinale fermentation broth: S1, Same as in Example 1; S2, Same as in Example 1; S3, fermentation time 60h, other aspects are the same as in Example 1.

[0062] Experimental tests were conducted on samples taken in Examples 1-15. Chronic inflammation model: DSS-induced mouse ulcerative colitis model; Experimental methods: (1) Experimental animals: C57BL / 6 strain mice, 6-8 weeks old, male, 6-10 mice per group. (2) DSS solution preparation: Sodium gluconate sulfate with a molecular weight of 36000-50000 Da, concentration of 2-3% (w / v). Method: Dissolve DSS powder in sterile drinking water of mice, protect from light, stir with a magnetic stirrer until fully dissolved, and use immediately after preparation; Modeling cycle: Free access to DSS water for 5 consecutive days; then switch back to normal drinking water for 7 days, repeating the cycle twice to simulate chronic relapse; Detection indicators: (1) Disease activity index (DAI): assessed daily from the third day of modeling, combining three scores: A. Weight loss (0 points: none; 1 point: 1-5%; 2 points: 5-10%; 3 points: 10-15%; 4 points: >15%) B. Stool condition (0 points: normal stool; 1 point: moist stool; 2 points: loose stool; 3 points: semi-loose stool; 4 points: diarrhea). C. Rectal bleeding / occult blood (0 points: none; 1 point: a small amount of visible rectal bleeding, mild; 2 points: a small amount of visible rectal bleeding, moderate; 3 points: a large amount of visible rectal bleeding; 4 points: a large amount of visible rectal bleeding, severe) D.DAI = (Weight Loss Score + Stool Characteristics Score + Blood in Stool) / 3 (2) Colonic histological scoring: At the end of the experiment, the length of the colon was measured by dissection. 0 points: No inflammation, no lesions, lesion range <1%, crypts intact; 1 point: Very mild inflammation, lesion depth to the submucosa, lesion range 1-25%, basal 1 / 3 of crypts destroyed; 2 points: Mild inflammation, lesion depth to the mucosa and muscularis propria, lesion range 26-50%, basal 2 / 3 of crypts destroyed; 3 points: Moderate inflammation, lesion depth to the muscularis propria, lesion range 51-75%, all crypts destroyed, but intact epithelium present; 4 points: Severe inflammation, lesion range 76-100%, lesion depth to the serosa, all crypts and intact epithelium destroyed.

[0063] Test results of Examples 1-15 Table 1 Test results of Examples 1-15

[0064] The blank control group consisted of mice that did not drink the Dendrobium officinale fermentation liquid composition; As shown in the table above, Examples 1-3 describe the effect of the amount of Dendrobium officinale fermentation liquid added on the anti-inflammatory effect of the composition. The results show that the anti-inflammatory effect of the composition is better with the increase of the amount of Dendrobium officinale fermentation liquid added. Examples 1 and 4-5 describe the effect of the amount of Eucommia ulmoides juice added on the anti-inflammatory effect of the composition. The anti-inflammatory effect of the composition increases with the increase of the amount of Eucommia ulmoides juice added. Examples 1 and 6-7 describe the effect of kiwifruit juice on the anti-inflammatory effect of the composition. The amount of kiwifruit juice added decreases slowly between 15 and 20 parts. Examples 1 and 8-9 describe the effect of sugarcane juice on the anti-inflammatory effect of the composition. Similar to the changes in kiwifruit juice, the amount added decreases slowly between 10 and 12 parts. Examples 1 and 10-15 describe the effect of polydextrose, lemon juice, and mint on the anti-inflammatory effect of the composition. The three have a small anti-inflammatory effect on the composition and only have a certain auxiliary ability. Through the blank control group (i.e. mice that did not drink the Dendrobium officinale fermentation liquid composition), it can be seen that the Dendrobium officinale fermentation liquid with anti-inflammatory effects of the present invention can effectively improve the resistance of mice to colitis, upregulate the expression of intestinal tissue connexin in mice, improve the anti-inflammatory effect of intestinal tissue, and increase the content of short-chain fatty acids in the mouse intestine.

[0065] Examples 16-30 describe the determination of total sugar and total phenol content in fermented Dendrobium officinale juice. Each experiment was repeated three times, and the results are expressed as mean ± standard deviation. Table 2 Test results of Examples 16-30

[0066] As shown in Table 2, and in Examples 1 and 16-19, the total phenol and total sugar content initially increased and then decreased with the change in the bacterial strain ratio, reaching an optimal level at 1:1. Regarding the change in total sugar, *Lactobacillus plantarum* produces fructooligosaccharides and extracellular polysaccharides; as their proportion increases, the total sugar content rises. Conversely, *Lactobacillus acidophilus* secretes lipases and proteases, which promote cell decomposition; when their proportion decreases, total sugar accumulates. When the bacterial strain ratio is optimal (e.g., 1:1), the pectinase and other enzymes secreted by the bacteria decompose the cell wall, releasing polyphenols into a free state, thus increasing the total phenol content. However, when the bacterial strain ratio is unbalanced (e.g., 1:3, 0.3:1), competition for space and nutrients among the bacteria leads to a decrease in the secretion efficiency of pectinase and cellulase, and some simple phenols are degraded as carbon sources, thus reducing the total phenol content.

[0067] As shown in Examples 1 and 20-23, the total sugar and total phenolic content initially increased and then decreased with increasing inoculum amount, reaching a peak at 5%. When the inoculum amount is too low (e.g., 1%), insufficient lactic acid bacteria reproduction leads to low bacterial density and low fermentation efficiency, making it difficult to effectively decompose Dendrobium officinale juice and release polysaccharides. When the inoculum amount increases from 1% to 5%, the metabolic activity of lactic acid bacteria intensifies, sugars are consumed, and fructooligosaccharides are synthesized, resulting in an increase in total sugar content. When the inoculum amount exceeds 5%, excessive inoculum leads to the consumption of nutrients, intensified nutrient competition among bacterial strains, and excessive accumulation of lactic acid, accelerating sugar degradation. When the inoculum amount increases from 1% to 5%, the large-scale reproduction of lactic acid bacteria promotes the decomposition of Dendrobium officinale cell walls by cellulase and other enzymes, releasing bound polyphenols. When the inoculum amount exceeds 5%, excessive competition for nutrients causes some simple phenols to be degraded as carbon sources, ultimately decreasing the total phenolic content.

[0068] As shown in Examples 1 and 24-27, the total sugar content initially increases and then decreases with increasing fermentation temperature. When the temperature rises from 31℃ to 37℃, the total sugar content gradually increases and reaches its peak at 37℃, then decreases as the temperature continues to rise. At lower temperatures, the enzyme production and activity of the bacteria are relatively low, limiting bacterial growth and resulting in a lower total sugar content in the fermentation broth. Excessively high temperatures destroy polysaccharide molecules and cause the precipitation of impurities such as proteins, and reduce or even kill the activity of lactic acid bacteria, thus leading to a decrease in total sugar content. The total phenol content shows the same trend. When the fermentation temperature rises from 31℃ to 37℃, the total phenol content gradually increases and then begins to decrease. This is because within this temperature range, the metabolic activity of lactic acid bacteria increases with increasing temperature, leading to increased secretion of enzymes such as pectinase, cellulase, and β-glucosidase, which promotes the destruction of plant cell wall structure and the release of phenolic and flavonoid compounds. As the temperature continues to rise, the metabolic activity of lactic acid bacteria is inhibited, thus reducing enzyme secretion efficiency and decreasing the total phenol content.

[0069] As shown in Examples 1 and 28-30, the total sugar content decreases continuously with prolonged fermentation time. Lactic acid bacteria consume sugar sources through metabolic activities to maintain growth and reproduction and produce metabolites. As fermentation time increases, lactic acid bacteria continuously utilize the sugar sources in the fermentation system. Simultaneously, during fermentation, *Lactobacillus plantarum* and *Lactobacillus acidophilus* continuously compete for nutrients and living space, leading to the death of some bacteria and limiting fermentation. Therefore, the total sugar content shows a gradual decreasing trend. The total phenolic content of *Dendrobium officinale* juice shows a trend of first increasing and then decreasing with fermentation time, but it remains higher than that of unfermented juice after fermentation. In the early stages of fermentation, as time increases, the metabolic activity of lactic acid bacteria gradually strengthens, and the enzymes secreted by them promote the release and synthesis of phenolic substances in *Dendrobium officinale* juice, thus the total phenolic content continues to rise. When the fermentation time reaches approximately 36 hours, the production of phenolic substances reaches its peak. At this time, lactic acid bacteria are in a suitable metabolic stage with high enzyme activity. As time continues to increase, the total phenolic content decreases, nutrients are consumed, lactic acid bacteria activity is inhibited, enzyme activity decreases, and metabolic activity weakens.

[0070] This invention relates to a fermentation broth composition of Dendrobium officinale, using fermentation broth of Dendrobium officinale as the main component, and adding Eucommia ulmoides juice, kiwi juice, and sugarcane juice as a base, along with polydextrose, lemon juice, and mint. Through single-factor experiments, the fermentation parameters of Dendrobium officinale were optimized, resulting in a method for preparing Dendrobium officinale fermentation broth. Experiments show that this composition has excellent anti-inflammatory effects and also exhibits excellent antioxidant properties. The method for preparing Dendrobium officinale fermentation broth of this invention ensures better fermentation results from the compound bacteria, leading to a more effective anti-inflammatory effect in the prepared Dendrobium officinale fermentation broth.

Claims

1. A Dendrobium officinale fermentation liquid composition with anti-inflammatory effects, characterized in that, By weight, 30-40 parts of Dendrobium officinale fermentation liquid, 15-25 parts of Eucommia ulmoides juice, 10-20 parts of kiwifruit juice, and 8-12 parts of sugarcane juice.

2. The Dendrobium officinale fermentation liquid composition with anti-inflammatory effects according to claim 1, characterized in that, The Dendrobium officinale fermentation liquid is derived from Dendrobium officinale juice through fermentation, and the flavonoid content of the Eucommia ulmoides juice is 3-4%.

3. The Dendrobium officinale fermentation liquid composition with anti-inflammatory effects according to claim 2, characterized in that, The fermentation bacteria in the Dendrobium officinale juice fermentation is a compound bacterial agent of Lactobacillus acidophilus and Lactobacillus plantarum.

4. The Dendrobium officinale fermentation liquid composition with anti-inflammatory effects according to claim 3, characterized in that, The ratio of Lactobacillus acidophilus to Lactobacillus plantarum is 0.3-3:1, and the inoculation amount is 1-9% of Dendrobium officinale juice.

5. The Dendrobium officinale fermentation liquid composition with anti-inflammatory effects according to claim 1, characterized in that, It also includes the following components, by weight: 3-5 parts polydextrose, 2-4 parts lemon juice, 0.5-1 part peppermint, and purified water to make up to 120 parts.

6. A method for preparing Dendrobium officinale fermentation broth, characterized in that, According to claim 2, the preparation of the Dendrobium officinale fermentation liquid composition with anti-inflammatory effects includes the following steps: S1. After washing the Dendrobium officinale stems, juice them using a high-speed blender at a material-to-liquid ratio of 1:20 (m / v), filter the juice, and then sterilize it for later use. S2. Cultivate the compound bacteria, and after three generations, wash three times with physiological saline for later use. S3. After inoculating the activated compound bacteria into the sterilized Dendrobium officinale juice for fermentation, Dendrobium officinale fermentation liquid is obtained.

7. The method for preparing Dendrobium officinale fermentation broth according to claim 6, wherein the sterilization process in step S1 is to sterilize at 121℃ for 10 min in an autoclave; and the cultivation in step S2 is to culture in MRS broth medium at 37℃ for 48 h, perform two subculturings, and then culture to the third generation after activation, obtaining an initial bacterial concentration of 2.6 × 10⁻⁶. 8 CFU / mL; the fermentation conditions for S3 are: fermentation temperature of 31-43℃ and fermentation time of 12-60h.

8. A method for preparing a Dendrobium officinale fermentation liquid composition, comprising the following steps: S1. Add the Dendrobium officinale fermentation liquid, Eucommia ulmoides juice, kiwi juice, and sugarcane juice to the mixing tank in proportion, stir, add polydextrose and continue stirring, then add lemon juice and mint pieces, and stir lightly to obtain a mixture. S2. The mixture is homogenized using a homogenizer, and then water is added to make up to 120 parts to obtain a preliminary Dendrobium officinale fermentation liquid composition. S3. The preliminary Dendrobium officinale fermentation liquid composition is sterilized to obtain a Dendrobium officinale fermentation liquid composition with anti-inflammatory effects.

9. A method for preparing a Dendrobium officinale fermentation liquid composition according to claim 8, wherein the kiwi juice and sugarcane juice in step S1 are fresh kiwi and sugarcane juices that have been washed, peeled, pressed, coarsely filtered, and sterilized. The sterilization temperature is 95°C and the sterilization time is 30s. The stirring rate in step S1 is 400-600 rpm and the stirring time is 10-20 min.

10. The method for preparing a Dendrobium officinale fermentation broth composition according to claim 8, wherein the pressure of the homogenizer in step S2 is 15-25 MPa, and the sterilization treatment in step S3 is 400-600 MPa for 5-10 min.

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