Sea cucumber aqueous extract anti-helicobacter pylori solid beverage and preparation method thereof

Abstract

The invention discloses a sea cucumber aqueous extract anti-helicobacter pylori solid beverage and a preparation method thereof. The solid beverage comprises the following substance components in parts by weight: 10-15 parts of a sea cucumber-hericium erinaceus aqueous extract mixture, 20-30 parts of resistant dextrin, 0.5-1 part of whey protein powder and 2-4 parts of xylitol. The preparation method comprises the following steps: obtaining the sea cucumber extract in a water extraction and alcohol precipitation manner; the hericium erinaceus extract is obtained in a water extraction and alcohol precipitation mode; the sea cucumber extract and the hericium erinaceus extract are redissolved, homogenized and then spray-dried, resistant dextrin, whey protein and xylitol are added, the mixture is mixed evenly and directly bagged, and the sea cucumber aqueous extract anti-helicobacter pylori solid beverage is obtained. The solid beverage disclosed by the invention has direct helicobacter pylori resisting activity in vitro, solves the problems of increased bacterial drug resistance, low eradication rate and large side effect in the existing quadruple therapy for treating helicobacter pylori, and can promote the deep processing of sea cucumbers and improve the additional value of sea cucumbers.

Description

Technical Field

[0001] The invention relates to a solid beverage and a preparation method thereof, in particular to a sea cucumber water extract anti-Helicobacter pylori solid beverage and a preparation method thereof. Background Art

[0002] Helicobacter pylori is a type of pathogenic bacteria that parasitizes the gastric mucosal epithelium of the human body. It has been identified as a Class I biological carcinogen by the World Health Organization and the International Agency for Research on Cancer. At present, the traditional treatment for Helicobacter pylori is quadruple therapy, which includes antibiotics. Long-term use will increase the drug resistance of Helicobacter pylori, reduce the eradication rate, and make it easy to get infected repeatedly. In addition, the use of multiple drugs can also cause serious toxic side effects such as vomiting, abdominal distension, diarrhea, and intestinal flora imbalance.

[0003] Sea cucumbers have been a nutritious tonic since ancient times. The ingredients in sea cucumbers, such as collagen, polysaccharides, polypeptides, etc., all have a variety of good biological activities. However, the sea cucumber industry is still mainly sold in the form of whole cucumbers, and deep processing is seriously insufficient. Therefore, it will be of great significance to develop a functional food with resistance to Helicobacter pylori based on sea cucumber water extracts. Summary of the invention

[0004] In order to solve the shortcomings of the above technology, the present invention provides a sea cucumber water extract anti-Helicobacter pylori solid beverage and a preparation method thereof.

[0005] In order to solve the above technical problems, the technical solution adopted by the present invention is: a sea cucumber water extract anti-Helicobacter pylori solid beverage, which includes the following material components in weight ratio:

[0006] 10-15 parts of sea cucumber-hericium erinaceus water extract mixture, 20-30 parts of resistant dextrin, 0.5-1 part of whey protein powder, and 2-4 parts of xylitol.

[0007] Preferably, the weight ratio of the sea cucumber water extract to the Hericium erinaceus water extract in the sea cucumber-Hericium erinaceus water extract mixture is 3:1.

[0008] A method for preparing a sea cucumber water extract anti-Helicobacter pylori solid beverage, the preparation method comprising:

[0009] The first step is to obtain sea cucumber extract by water extraction and alcohol precipitation;

[0010] The second step is to obtain Hericium erinaceus extract by water extraction and alcohol precipitation;

[0011] The third step is to re-dissolve the sea cucumber extract and Hericium erinaceus extract, homogenize and then spray-dry, add resistant dextrin, whey protein and xylitol, mix well and directly bag to obtain a sea cucumber water extract anti-Helicobacter pylori solid beverage.

[0012] Preferably, the preparation method comprises the following specific steps:

[0013] S1, freeze-drying and crushing the whole sea cucumber to obtain sea cucumber powder;

[0014] S2, adding sodium acetate buffer and papain to the sea cucumber powder for enzymatic extraction, separating the solid and liquid after extraction, and collecting the supernatant;

[0015] S3, add cetylpyridinium chloride solution to the supernatant, react at room temperature for 10-30 hours, and collect the precipitate;

[0016] S4, dissolving the precipitate in a NaCl solution-ethanol mixed solution, adding ethanol, mixing and standing, and collecting the precipitate;

[0017] S5, re-dissolving the precipitate collected in S4, desalting, concentrating and freeze-drying to obtain a sea cucumber water extract;

[0018] S6. crushing the dried Hericium erinaceus;

[0019] S7, dissolving Hericium erinaceus powder in water, performing extraction, performing solid-liquid separation after extraction, and collecting the supernatant;

[0020] S8, adding ethanol to the Hericium erinaceus extraction supernatant, mixing and standing, collecting the precipitate, and obtaining the Hericium erinaceus water extract;

[0021] S9, mixing the sea cucumber water extract obtained in S5 and the Hericium erinaceus water extract obtained in S8 in a weight ratio of 3:1, re-dissolving, homogenizing and spray-drying to obtain a sea cucumber-Hericium erinaceus water extract mixture;

[0022] S10. 10-15 parts of the sea cucumber-hericium erinaceus water extract mixture, 20-30 parts of resistant dextrin, 0.5-1 part of whey protein powder, and 2-4 parts of xylitol are mixed according to a weight ratio to obtain a sea cucumber water extract anti-Helicobacter pylori solid beverage.

[0023] Preferably, in S2, the concentration of the sodium acetate buffer is 0.1-0.3 mol / L; the material-liquid ratio of the sea cucumber powder to the sodium acetate buffer is 1:20-30 g / mL;

[0024] The enzymatic activity of papain is 700-900U / mg; 0.04-0.08g of papain is used for each gram of sea cucumber powder.

[0025] Preferably, in S2, the temperature for enzymatic extraction of sea cucumber powder is 50-70°C and the time is 20-30h;

[0026] In S7, the extraction temperature of Hericium erinaceus is 50-70°C.

[0027] Preferably, in S3, the mass volume ratio of the cetylpyridinium chloride solution is 10%; the solid-liquid ratio of the sea cucumber powder to the cetylpyridinium chloride solution is 1:1.6-2 g / mL.

[0028] Preferably, in S4, in the NaCl solution-ethanol mixed solution, the volume ratio of the NaCl solution to the ethanol is 100:15;

[0029] The concentration of the NaCl solution is 2-3 mol / L; the volume concentration of the ethanol is 95%.

[0030] Preferably, in S4 and S8, when the mixed solution and ethanol are mixed and allowed to stand, the volume ratio of the two is mixed solution: ethanol = 1:3-6; the standing temperature is 4-8°C, and the standing time is 12-24h.

[0031] Preferably, in S5, the desalination is ultrafiltration desalination with a molecular weight cut-off of 5000Da.

[0032] The present invention provides a solid beverage mainly composed of sea cucumber water extract and added with hericium erinaceus water extract having stomach protecting and stomach-protecting effects, and a preparation method thereof. The sea cucumber water extract anti-Helicobacter pylori solid beverage obtained by the preparation method of the present invention has direct anti-Helicobacter pylori activity in vitro, solving the problems of increased bacterial resistance, low eradication rate and large side effects in the existing quadruple therapy for the treatment of Helicobacter pylori, and can promote the deep processing of sea cucumbers and improve the added value of sea cucumbers. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 Schematic diagram of Helicobacter pylori culture in an embodiment of the present invention.

[0034] Figure 2 It is an electron microscope image of Helicobacter pylori in the embodiment of the present invention.

[0035] Figure 3 This is a graph showing the results of the anti-Helicobacter pylori inhibition zone experiment of the solid beverage 1 in the embodiment of the present invention.

[0036] Figure 4 This is a graph showing the results of the anti-Helicobacter pylori inhibition zone experiment of the solid beverage 2 in the embodiment of the present invention. DETAILED DESCRIPTION

[0037] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of this application.

[0038] Wherein, the reagents or raw materials used in the following examples are all commercially available or homemade.

[0039] The invention discloses a sea cucumber water extract anti-Helicobacter pylori solid beverage, which comprises the following material components in weight proportion: 10-15 parts of sea cucumber-hericium erinaceus water extract mixture, 20-30 parts of resistant dextrin, 0.5-1 part of whey protein powder and 2-4 parts of xylitol.

[0040] Among them, the sea cucumber-Hericium erinaceus water extract mixture is a mixture of sea cucumber water extract and Hericium erinaceus water extract, and the weight ratio of the two is 3:1.

[0041] The polysaccharide components contained in sea cucumbers have the effects of sterilization, anti-inflammatory, and repairing gastric mucosa, and have a certain auxiliary inhibitory effect on pathogens such as Helicobacter pylori. The present invention combines the sea cucumber water extract with the Hericium erinaceus water extract which has a stomach-nourishing effect, so that the solid beverage has direct anti-Helicobacter pylori activity in vitro.

[0042] The sea cucumber extract and Hericium erinaceus extract are then re-dissolved, homogenized and spray-dried, and resistant dextrin, whey protein and xylitol are added, mixed and directly bagged to obtain a sea cucumber water extract anti-Helicobacter pylori solid beverage.

[0043] The preparation method of the sea cucumber water extract anti-Helicobacter pylori solid beverage of the present invention comprises the following specific steps:

[0044] S1, freeze-drying and crushing the whole sea cucumber to obtain sea cucumber powder;

[0045] S2. Sodium acetate buffer and papain are added to the sea cucumber powder for enzymatic extraction, solid-liquid separation is performed after extraction, and the supernatant is collected; the sea cucumber powder is enzymatically hydrolyzed with papain, and the connecting bonds between some sea cucumber molecules are broken, thereby destroying their molecular structure, achieving the purpose of peeling, and at the same time, no odor is generated, and no loss of nutrients is caused, so as to better maintain the nutritional value of the sea cucumber.

[0046] S3, add cetylpyridinium chloride solution to the supernatant, react at room temperature for 10-30 hours, and collect the precipitate;

[0047] S4, dissolving the precipitate in a NaCl solution-ethanol mixed solution, adding ethanol, mixing and standing, and collecting the precipitate;

[0048] S5, re-dissolving the precipitate collected in S4, desalting, concentrating and freeze-drying to obtain a sea cucumber water extract;

[0049] S6. crushing the dried Hericium erinaceus;

[0050] S7, dissolving Hericium erinaceus powder in water, performing extraction, performing solid-liquid separation after extraction, and collecting the supernatant;

[0051] S8, adding ethanol to the Hericium erinaceus extraction supernatant, mixing and standing, collecting the precipitate, and obtaining the Hericium erinaceus water extract;

[0052] S9, mixing the sea cucumber water extract obtained in S5 and the Hericium erinaceus water extract obtained in S8 in a weight ratio of 3:1, re-dissolving, homogenizing and spray-drying to obtain a sea cucumber-Hericium erinaceus water extract mixture;

[0053] S10. 10-15 parts of the sea cucumber-hericium erinaceus water extract mixture, 20-30 parts of resistant dextrin, 0.5-1 part of whey protein powder, and 2-4 parts of xylitol are mixed according to a weight ratio to obtain a sea cucumber water extract anti-Helicobacter pylori solid beverage.

[0054] In some embodiments of the present invention, in S2, the concentration of sodium acetate buffer is 0.1-0.3 mol / L; the material-liquid ratio of sea cucumber powder to sodium acetate buffer is 1:20-30 g / mL;

[0055] In some embodiments of the present invention, in S2, the papain activity is 700-900 U / mg; and 0.04-0.08 g of papain is used for treatment per gram of sea cucumber powder.

[0056] In some embodiments of the present invention, in S2, the temperature of enzymatic extraction of sea cucumber powder is 50-70°C and the time is 20-30h; the extraction temperature of Hericium erinaceus is 50-70°C.

[0057] In some embodiments of the present invention, in S3, the mass volume ratio of the cetylpyridinium chloride solution is 10%; the solid-liquid ratio of the sea cucumber powder to the cetylpyridinium chloride solution is 1:1.6-2 g / mL.

[0058] In some embodiments of the present invention, in S4, in the NaCl solution-ethanol mixed solution, the volume ratio of NaCl solution to ethanol is 100:15; the concentration of NaCl solution is 2-3 mol / L; and the concentration of ethanol is 95%, by volume.

[0059] In some embodiments of the present invention, in S4 and S8, when the mixed solution and ethanol are mixed and allowed to stand, the volume ratio of the two is mixed solution: ethanol = 1:3-6; the standing temperature is 4-8°C, and the standing time is 12-24h.

[0060] In some embodiments of the present invention, in S5, the desalination is ultrafiltration desalination with a molecular weight cut-off of 5000Da.

[0061] Example

[0062] The method disclosed in the present invention is used to prepare a sea cucumber water extract anti-Helicobacter pylori solid beverage, comprising the following steps:

[0063] S1, freeze-drying and crushing the whole fresh sea cucumber to obtain sea cucumber powder;

[0064] S2, add 0.1mol / L sodium acetate buffer to 100g sea cucumber powder, and the solid-liquid ratio of the crushed sea cucumber powder to the sodium acetate buffer is 1:20g / mL; 8g papain (800U / mg), stir and react at 60°C for 24h, centrifuge, and collect the supernatant;

[0065] S3, adding 10% (w / v) cetylpyridinium chloride (CPC) solution to the supernatant, crushing the obtained sea cucumber powder to the CPC solution in a solid-liquid ratio of 1:1.6 g / mL, stirring and standing for 24 hours, centrifuging, and collecting the precipitate;

[0066] S4. The precipitate obtained in S3 was fully dissolved in a mixed solution of 3 mol / L NaCl solution: 95% (v / v) ethanol = 100:15 (v / v), and 3 times the volume of the mixed solution of 95% (v / v) ethanol solution was added. The mixture was allowed to stand at 4°C for 24 h, and the precipitate was collected by centrifugation.

[0067] S5, re-dissolving the precipitate to a concentration of about 10 mg / mL, desalting by ultrafiltration with a molecular weight cut-off of 5000 Da; concentrating and freeze-drying after desalting to obtain a sea cucumber water extract;

[0068] S6, crushing the dried Hericium erinaceus to obtain Hericium erinaceus powder;

[0069] S7, dissolving Hericium erinaceus powder in water with a solid-liquid ratio of 1:20 g / mL; extracting at 60° C., centrifuging, and collecting the supernatant;

[0070] S8, adding 4 times the volume of ethanol to the supernatant, leaving it to stand at 4°C overnight, collecting the precipitate to obtain the Hericium erinaceus water extract;

[0071] S9, mixing and re-dissolving the sea cucumber water extract and the Hericium erinaceus water extract in a weight ratio of 3:1, homogenizing and spray-drying to obtain a sea cucumber-Hericium erinaceus water extract mixture;

[0072] S10, mixing 15 parts of the sea cucumber-hericium erinaceus aqueous extract mixture, 20 parts of resistant dextrin, 0.5 parts of whey protein powder, and 2 parts of xylitol according to a weight ratio to obtain a solid beverage 1;

[0073] Solid beverage 2 was obtained by mixing 10 parts of sea cucumber-hericium erinaceus water extract mixture, 30 parts of resistant dextrin, 1 part of whey protein powder and 4 parts of xylitol according to the weight ratio.

[0074] The anti-Helicobacter pylori activity of the solid beverage 1 and the solid beverage 2 was measured, specifically:

[0075] Cultivation of Helicobacter pylori: Helicobacter pylori (purchased from Shangcheng Beina Chuanglian Biotechnology Co., Ltd.) was revived in Columbia blood agar medium containing 7% defibrinated sheep blood and cultured in a three-gas incubator (5% O2, 10% CO2, 85% N2) for 72 h.

[0076] Characteristics of Helicobacter pylori colonies Figure 1 As shown in the figure, a round, raised, transparent, needle-like colony with a diameter of about 1 mm appears on the culture medium. When the concentration is high, a layer of translucent bacterial lawn is formed. Take another plate of Helicobacter pylori for Gram staining, as shown in the figure. Figure 2 As shown, microscopic observation of Helicobacter pylori shows Gram-negative, S-shaped, arc-shaped or short rod-shaped, curved bacteria.

[0077] Sea cucumber water extract solid beverage antibacterial zone experiment:

[0078] The sea cucumber water extract solid beverage 1 and solid beverage 2 were prepared into 40 mg / mL, and amoxicillin was prepared into 20 ug / mL, and set aside. Sterile water was used as a blank control and amoxicillin was used as a positive control.

[0079] Scrape an appropriate amount of Helicobacter pylori, place it in Columbia agar medium containing 10% sterile defibrinated sheep blood, pour it into a plate, and after the medium solidifies, use sterile tweezers to stick a 6mm diameter sterile filter paper on the plate and press it gently. Place 5 filter paper pieces on each plate, drop 5μL sterile water (blank control, 2 pieces), amoxicillin solution (positive control) and 2 solid beverage groups, respectively, put the plate into a three-gas incubator, and take it out for observation after 72h of incubation.

[0080] The results are as follows Figure 3 and Figure 4 As shown, at the same time, Table 1 shows the results of the anti-Helicobacter pylori inhibition zone diameter. Within the experimental dose and limited time range, the blank control group had no inhibition zone, and the positive drug control group amoxicillin inhibition zone diameter was ≥15mm, indicating that amoxicillin has a strong direct inhibition activity against Helicobacter pylori. The inhibition zone diameters of sea cucumber water extract solid beverages 1 and 2 were not as good as those of amoxicillin in the positive control group, but both showed the ability to directly inhibit Helicobacter pylori activity.

[0081] Table 1 Diameter of the inhibition zone of solid beverage against Helicobacter pylori

[0082]

[0083] Note: a The diameter of the inhibition zone includes the diameter of the filter paper: 6 mm

[0084] The above implementation modes are not limitations of the present invention, and the present invention is not limited to the above examples. Changes, modifications, additions or substitutions made by technicians in this technical field within the scope of the technical solution of the present invention also belong to the protection scope of the present invention.

Claims

1. A sea cucumber water extract anti-Helicobacter pylori solid beverage, characterized in that: It includes the following material components in weight ratio: 10-15 parts of sea cucumber-hericium erinaceus water extract mixture, 20-30 parts of resistant dextrin, 0.5-1 part of whey protein powder, and 2-4 parts of xylitol.

2. The sea cucumber water extract anti-Helicobacter pylori solid beverage according to claim 1, characterized in that: The weight ratio of the sea cucumber water extract to the Hericium erinaceus water extract in the sea cucumber-Hericium erinaceus water extract mixture is 3:

1.

3. A method for preparing a sea cucumber water extract anti-Helicobacter pylori solid beverage according to any one of claims 1 to 2, characterized in that: The preparation method comprises: The first step is to obtain sea cucumber extract by water extraction and alcohol precipitation; The second step is to obtain Hericium erinaceus extract by water extraction and alcohol precipitation; The third step is to re-dissolve the sea cucumber extract and Hericium erinaceus extract, homogenize and then spray-dry, add resistant dextrin, whey protein and xylitol, mix well and directly bag to obtain a sea cucumber water extract anti-Helicobacter pylori solid beverage.

4. The method for preparing the sea cucumber water extract anti-Helicobacter pylori solid beverage according to claim 3, characterized in that: The specific steps of the preparation method are as follows: S1, freeze-drying and crushing the whole sea cucumber to obtain sea cucumber powder; S2, adding sodium acetate buffer and papain to the sea cucumber powder for enzymatic extraction, separating the solid and liquid after extraction, and collecting the supernatant; S3, add cetylpyridinium chloride solution to the supernatant, react at room temperature for 10-30h, and collect the precipitate; S4, dissolving the precipitate in a NaCl solution-ethanol mixed solution, adding ethanol, mixing and standing, and collecting the precipitate; S5, re-dissolving the precipitate collected in S4, desalting, concentrating and freeze-drying to obtain a sea cucumber water extract; S6. crushing the dried Hericium erinaceus; S7, dissolving Hericium erinaceus powder in water, performing extraction, performing solid-liquid separation after extraction, and collecting the supernatant; S8, adding ethanol to the Hericium erinaceus extraction supernatant, mixing and standing, collecting the precipitate, and obtaining the Hericium erinaceus water extract; S9, mixing the sea cucumber water extract obtained in S5 and the Hericium erinaceus water extract obtained in S8 in a weight ratio of 3:1, re-dissolving, homogenizing and spray-drying to obtain a sea cucumber-Hericium erinaceus water extract mixture; S10. 10-15 parts of the sea cucumber-hericium erinaceus water extract mixture, 20-30 parts of resistant dextrin, 0.5-1 part of whey protein powder, and 2-4 parts of xylitol are mixed according to a weight ratio to obtain a sea cucumber water extract anti-Helicobacter pylori solid beverage.

5. The method for preparing the sea cucumber water extract anti-Helicobacter pylori solid beverage according to claim 4, characterized in that: In S2, the concentration of the sodium acetate buffer is 0.1-0.3 mol / L; the material-liquid ratio of sea cucumber powder to sodium acetate buffer is 1:20-30 g / mL; The papain activity is 700-900 U / mg; 0.04-0.08 g of papain is used for each gram of sea cucumber powder.

6. The method for preparing the sea cucumber water extract anti-Helicobacter pylori solid beverage according to claim 4, characterized in that: In S2, the temperature of enzymatic extraction of sea cucumber powder is 50-70°C and the time is 20-30h; In S7, the extraction temperature of Hericium erinaceus is 50-70°C.

7. The method for preparing the sea cucumber water extract anti-Helicobacter pylori solid beverage according to claim 4, characterized in that: In S3, the mass volume ratio of the cetylpyridinium chloride solution is 10%; the solid-liquid ratio of the sea cucumber powder to the cetylpyridinium chloride solution is 1:1.6-2 g / mL.

8. The method for preparing the sea cucumber water extract anti-Helicobacter pylori solid beverage according to claim 4, characterized in that: In S4, in the NaCl solution-ethanol mixed solution, the volume ratio of NaCl solution to ethanol is 100:15; The concentration of the NaCl solution is 2-3 mol / L; the volume concentration of the ethanol is 95%.

9. The method for preparing the sea cucumber water extract anti-Helicobacter pylori solid beverage according to claim 4, characterized in that: In S4 and S8, when the mixed solution and ethanol are mixed and allowed to stand, the volume ratio of the two is mixed solution: ethanol = 1:3-6; the standing temperature is 4-8°C, and the standing time is 12-24h.

10. The method for preparing the sea cucumber water extract anti-Helicobacter pylori solid beverage according to claim 4, characterized in that: In S5, the desalination is ultrafiltration desalination with a molecular weight cut-off of 5000Da.

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