Extraction process of jellyfish extract

Abstract

The invention relates to the technical field of collagen and mucoprotein extraction, and discloses a jellyfish extract extraction process which comprises the following steps: (1) pretreatment: mainly comprising the steps of cleaning, desalting, acid and alkali treatment, homogenization, enzymolysis, centrifugation and the like, and the specific operation is performed according to the patent (application number: CN118255872A); (2) ultrafiltration and concentration: separating the supernate obtained by pretreatment, and retaining a concentrated solution; and (3) solid drying: freeze-drying the concentrated solution to obtain a jellyfish extract solid product rich in collagen and mucoprotein. The jellyfish extract prepared by the method is a white or faint yellow solid, has no peculiar smell, is rich in collagen and mucoprotein, and is widely applied to the field of cosmetics.

Description

Technical Field

[0001] The invention relates to the technical field of collagen and mucin extraction, in particular to an extraction process of jellyfish extract. Background Art

[0002] At present, there are many kinds of cosmetics, and their various formulas also dazzle us. The use effect of cosmetics will be affected by many factors such as product formula, concentration and personal skin quality. A jellyfish extract provided by the present invention is suitable for the field of cosmetics, and this jellyfish extract contains collagen and mucin at the same time. Collagen has the characteristics of low immunogenicity, good biocompatibility, and high biological activity. It is used in the field of cosmetics and has the effects of anti-wrinkle, firming, moisturizing, whitening and freckle removal, anti-oxidation, soothing, and repairing; mucin has low biological rejection and has great utilization value to the human body. It helps to promote the production of collagen and elastin, increase skin elasticity, reduce wrinkles, and make the skin firmer and smoother. It can be seen that the jellyfish extract prepared by the present invention is a preferred product as a cosmetic raw material.

[0003] In the past, Patent 1 disclosed a method for graded extraction of mucin and collagen, but its extraction rate was low, and organic solvents such as ethanol were used, resulting in high costs and unsuitability for large-scale production. In addition, Patent 2 (application number: CN118255872A) proposed a jellyfish collagen and its preparation process. Document 1 studied the extraction and structural characteristics of jellyfish collagen, and Document 2 studied the extraction of jellyfish collagen and its physicochemical properties. However, none of the three articles involved the extraction of mucin. Document 3 studied the extraction, process purification, physicochemical properties, and biological activity of glycoproteins in the jellyfish mouth arm, but did not mention the extraction process of jellyfish collagen. Document 4 has pointed out that the extraction methods of jellyfish polysaccharide include: hot water extraction, protease extraction, alkali extraction of polysaccharide, alcohol precipitation, etc., and it is clearly pointed out that hot water extraction is the most commonly used method for extracting jellyfish polysaccharide, but its higher temperature can accelerate the extraction rate of polysaccharide, and at the same time, it will denature some proteins in the jellyfish body and reduce the protein content in the jellyfish polysaccharide extract. There is a problem that some glycosidic bonds of alkali-extracted polysaccharide are degraded due to alkali treatment. Therefore, in order to solve the existing technical problems, the present invention combines enzyme method with membrane separation equipment to ensure that the raw materials are fully reacted, and uses 30kDa membrane for ultrafiltration, which increases the extraction rate of mucin and collagen, and does not use organic solvents such as ethanol, which is economical and suitable for large-scale production.

[0004] Therefore, we propose an extraction process of jellyfish extract. Summary of the invention

[0005] The present invention mainly solves the technical problems existing in the above-mentioned prior art and provides an extraction process of jellyfish extract.

[0006] In order to achieve the above object, the present invention adopts the following technical scheme, a jellyfish extract extraction process, comprising the following steps:

[0007] (1) Pretreatment: mainly includes: cleaning, desalting, acid-base treatment, homogenization, enzymatic hydrolysis, centrifugation and other steps. The specific operations are carried out according to the patent (application number: CN118255872A).

[0008] (2) Ultrafiltration concentration: Separate the supernatant obtained from the pretreatment and retain the concentrated solution.

[0009] (3) Solid drying: The concentrated solution is freeze-dried to obtain a solid product of jellyfish extract rich in collagen and mucin.

[0010] As a preferred technical solution of the present invention, the (2) ultrafiltration concentration: using an ultrafiltration membrane with a molecular weight cutoff of 30 kDa to separate the supernatant obtained from the pretreatment, and retaining the concentrated solution.

[0011] The present invention provides a jellyfish extract extraction process. It has the following beneficial effects

[0012] 1. The jellyfish extract provided by the present invention is rich in collagen and mucin, has multiple functions such as moisturizing, nourishing the skin, anti-oxidation, whitening and anti-aging, and is widely used in the field of cosmetics.

[0013] 2. The present invention adopts an ultrafiltration membrane with a molecular weight cutoff of 30 kDa to simultaneously extract collagen and mucin from jellyfish, thereby reducing production costs and avoiding environmental pollution and waste of jellyfish mucin resources. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] The structures, proportions, sizes, etc. illustrated in this specification are only used to match the contents disclosed in the specification for people familiar with this technology to understand and read, and are not used to limit the conditions under which the present invention can be implemented, so they have no substantial technical significance.

[0015] Figure 1 This is the electrophoresis diagram of jellyfish collagen;

[0016] Figure 2 This is the electrophoresis diagram of jellyfish extract. DETAILED DESCRIPTION

[0017] Example 1

[0018] See also Figure 1 , a process for extracting mucin from jellyfish, comprising the following steps:

[0019] (1) Thawing: Thaw the jellyfish naturally and set aside.

[0020] (2) Grinding and cell disruption: Grind the thawed jellyfish and then perform ultrasonic cell disruption.

[0021] (3) Extraction: Add NaCl solution to the jellyfish treated as above and extract at 4°C.

[0022] (4) Centrifugation: Centrifuge the above solution at 4000 r / min for 20 min and collect the supernatant.

[0023] (5) Alcohol precipitation: The supernatant is precipitated with alcohol in an ice bath.

[0024] (6) Dialysis: Dissolve the precipitate in distilled water and dialyze in running water for 24 h, then in distilled water at 4°C for 24 h (change the distilled water every 4 h).

[0025] (7) Vacuum freeze drying: The dialyzed liquid is freeze dried to obtain jellyfish mucin with a molecular weight of 41.53 kDa.

[0026] Example 2

[0027] A jellyfish collagen protein and a preparation process thereof, comprising the following steps:

[0028] (1) Cleaning: Place the fresh jellyfish in a cleaning bubbler and add clean water for cleaning.

[0029] (2) Acid swelling: Cut the cleaned sample with skin into small uniform pieces, add 0.25 mol / L citric acid solution at a ratio of w:v=1:3, soak for 6 h, drain and set aside.

[0030] (3) Homogenization: Use a colloid mill to homogenize the above sample to obtain a colloidal sample.

[0031] (4) Enzymatic hydrolysis: After homogenization, the pH of the sample was adjusted to below 2, 0.3 wt% pepsin was added, and the enzymatic hydrolysis reaction was continuously stirred at 9°C for 24 h.

[0032] (5) Centrifugation: The above enzymatic hydrolysate was centrifuged at 9700 rpm in a butterfly centrifuge, and then centrifuged at 14000 rpm in a tubular centrifuge, and the supernatant was retained.

[0033] (6) Microfiltration: The supernatant was passed through a 0.22 μm PES microfiltration membrane to remove bacteria.

[0034] (7) Ultrafiltration and concentration: Use polyethersulfone ultrafiltration membrane filter to intercept and concentrate collagen macromolecules with a molecular weight of more than 80 kDa. Monitor the conductivity of the ultrafiltrate (located on the original solution side) in real time and stop when the conductivity is as low as 24 μs / cm.

[0035] (8) Freeze drying: The ultrafiltration concentrate was placed in a freeze dryer for freeze drying, with the vacuum degree set to 5 Pa and the cold trap temperature set to -45°C to obtain a collagen solid product with a molecular weight of 135 kDa. Figure 1 .

[0036] Example 3

[0037] A process for extracting jellyfish extract comprises the following steps:

[0038] (1) Pre-treatment:

[0039] S1: Cleaning: Put the fresh jellyfish into clean water for cleaning to remove impurities;

[0040] S2: Desalination: Soak the salted jellyfish in clean water or use running water to wash and desalinate. Take a certain volume of soaking liquid and add silver nitrate until there is no white turbidity, which means the desalination is successful;

[0041] S3: Acid-base treatment: Take the cleaned / desalted fresh / salted jellyfish samples and soak them in a certain proportion of acid / base for 3h to 12h;

[0042] S4: homogenization: homogenizing the processed sample to obtain a jellyfish homogenate;

[0043] S5: Enzymatic hydrolysis: Take the homogenized sample, add acid solution according to the ratio of W:V=1:0.2~2, add 1~3.0wt% enzyme of the raw material, and perform enzymatic hydrolysis for 8h~48h at a temperature of 4℃~10℃;

[0044] S6: Centrifugation: Centrifuge the above enzymatic hydrolysate at 12000 r / min and retain the supernatant;

[0045] (2) Ultrafiltration concentration: Use an ultrafiltration membrane with a molecular weight cutoff of 30 kDa to separate the supernatant and retain the concentrate;

[0046] (3) Solid drying: The concentrated solutions are mixed and freeze-dried to obtain a solid product of jellyfish extract rich in collagen and mucin.

[0047] Example 4

[0048] Preparation of fresh jellyfish extract:

[0049] A process for extracting jellyfish extract comprises the following steps:

[0050] (1) Pre-treatment:

[0051] S1: Cleaning: Put the fresh jellyfish into clean water for cleaning to remove impurities;

[0052] S2: Acid treatment: Take the cleaned fresh jellyfish sample and soak it in 0.5M citric acid at 4°C for 12h according to W:V=1:3;

[0053] S4: homogenization: homogenize the treated sample for 2 to 5 minutes to obtain a jellyfish homogenate;

[0054] S5: Enzymatic hydrolysis: Take the homogenized sample, add 0.5M citric acid according to the ratio of W:V=1:0.5, add 3.0wt% of the enzyme in the raw material amount, and perform enzymatic hydrolysis for 24h at a temperature of 8°C;

[0055] S6: Centrifugation: Centrifuge the above enzymatic hydrolysate at 12000 r / min and retain the supernatant;

[0056] (2) Ultrafiltration concentration: Use an ultrafiltration membrane with a molecular weight cutoff of 30 kDa to separate the supernatant and retain the concentrate;

[0057] (3) Solid drying: freeze-drying or spray-drying the concentrated solution to obtain a solid product of fresh jellyfish extract suitable for the cosmetics field, which is rich in collagen and mucin.

[0058] Example 5

[0059] Preparation of salted jellyfish extract:

[0060] (1) Pre-treatment:

[0061] S1: Desalination: Soak the salted jellyfish in clean water or use running water to wash and desalinate. Take a certain volume of soaking liquid and add silver nitrate until there is no white turbidity, which means the desalination is successful;

[0062] S2: Alkaline treatment: Take the cleaned and desalted salted jellyfish sample and soak it in 0.1M sodium hydroxide at 25°C for 6h according to W:V=1:6;

[0063] S4: Homogenization: Wash the sample treated with alkali with deionized water until it is neutral and then homogenize for 2 to 5 minutes to obtain a jellyfish homogenate;

[0064] S5: Enzymatic hydrolysis: Take the homogenized sample, add 0.5M citric acid according to the ratio of W:V=1:0.5, add 3.0wt% of the enzyme in the raw material amount, and perform enzymatic hydrolysis for 24h at a temperature of 8°C;

[0065] S6: Centrifugation: Centrifuge the above enzymatic hydrolysate at 12000 r / min and retain the supernatant;

[0066] (2) Ultrafiltration concentration: Use an ultrafiltration membrane with a molecular weight cutoff of 30 kDa to separate the supernatant and retain the concentrate;

[0067] (3) Solid drying: freeze-drying or spray-drying the concentrated solution to obtain a solid product of salted jellyfish extract suitable for the cosmetics field, which is rich in collagen and mucin.

[0068] Example 6

[0069] Determination of Hydroxyproline and Total Sugar Content in Jellyfish Extract:

[0070] (1) Determination of Hydroxyproline Content

[0071] Detection method:

[0072] ① Take 10-15 mg of fresh / salted jellyfish extract solid, add 3 mL of 6 mol / L hydrochloric acid solution, hydrolyze at 130℃ for 4 hours, take out and cool. Add a drop of methyl red indicator, adjust to pH=6, and make up to 100 mL. This solution is the sample test solution.

[0073] ② With deionized water as blank, take 1mL of the above test solution in a test tube, add 1mL of citric acid buffer and 1mL of chloramine T, shake well, oxidize at room temperature for 10min, add 1mL of 27% perchloric acid, shake well, react at room temperature for 10min, add 1mL of coloring agent, shake well, and place in a 65℃ water bath for color development for 20min. After cooling, measure the absorbance at 560nm. The test results are shown in Table 1:

[0074] Table 1 Hydroxyproline detection results

[0075]

[0076] (2) Determination of total sugar content

[0077] Detection Methods

[0078] ① Weigh 0.25g~0.4g of fresh / salted jellyfish extract solid sample, carefully pour the sample into a 100mL conical flask, add 20mL water and 6mL concentrated hydrochloric acid. Install a condensation reflux device and place it in a 100℃ water bath for hydrolysis for 3h. Cool to room temperature and filter, then wash the filter residue with distilled water, combine the filtrate and washing liquid, and dilute to 100mL with water. This solution is the sample test solution.

[0079] ② Accurately pipette 0.2mL of the sample test solution into a 10mL stoppered test tube, fill it up to 1.0mL with distilled water, add 1mL of phenol solution, and then quickly add 5mL of concentrated sulfuric acid. Let the reaction solution stand for 10 minutes, vortex, and react the test tube in a 30℃ water bath for 20 minutes. Adjust to zero with a blank solution, and measure the absorbance at 490nm. The test results are shown in Table 2:

[0080] Table 2 Total sugar test results

[0081]

[0082] Example 7

[0083] Determination of jellyfish extract by electrophoresis

[0084] Detection method:

[0085] ① Glue making: Clean the gel plates and place them in the single-hole gel making machine. Use paper to absorb the remaining water. First, prepare the separation gel in a certain proportion, mix well, and add it to the prepared gel plate with a pipette. Finally, add isopropanol on the separation gel to keep the separation gel level. Leave it for 30 minutes to solidify. Clean the solidified separation gel, absorb the remaining water, prepare the concentrated gel in a certain proportion, add it to the gel plate with a pipette, and immediately insert the comb. It can be used after about 30 minutes.

[0086] ② Sample treatment: Weigh 4 mg of sample, add 1 mL of acetic acid-sodium acetate buffer solution, vortex for 2 min, extract at 4 °C for 2 h, vortex again until the sample is dissolved, centrifuge at 10000r for 5 min, take the supernatant, mix the sample according to 4:1 (sample: loading), then boil in water bath for 5 min, cool to room temperature and set aside.

[0087] ③ Loading: First add 10μL protein marker, then add 20μL treated jellyfish extract in sequence.

[0088] ④Electrophoresis: electrophoresis was performed at 120 V for 65 min.

[0089] ⑤ Staining: Carefully remove the gel after electrophoresis, put it into a gel box, add Coomassie Brilliant Blue staining solution, and stain on a shaker for 1 hour.

[0090] ⑥ Decolorization: Pour out the staining solution, add Coomassie Brilliant Blue R-250 decolorization solution, decolorize for two hours, and observe the results under the irradiation of the UV gel. Figure 2 .

[0091] In summary, the jellyfish extract prepared by the present invention is rich in collagen and mucin, can be widely used in the field of cosmetics, and has the functions of moisturizing, nourishing the skin, anti-oxidation, whitening and anti-aging, etc. It not only reduces the production cost, but also avoids environmental pollution and waste of resources.

[0092] The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention. It should be understood by those skilled in the art that the present invention is not limited by the above embodiments, and the above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, and these changes and improvements all fall within the scope of the present invention to be protected.

Claims

1. A process for extracting jellyfish extract, characterized in that: The following steps are involved: (1) Pretreatment: mainly includes: cleaning, desalting, acid-base treatment, homogenization, enzymatic hydrolysis, centrifugation and other steps. The specific operations are carried out according to the patent (application number: CN118255872A). (2) Ultrafiltration concentration: Separate the supernatant obtained from the pretreatment and retain the concentrated solution. (3) Solid drying: The concentrated solution is freeze-dried to obtain a solid product of jellyfish extract rich in collagen and mucin.

2. According to claim 1 (2) ultrafiltration concentration: using an ultrafiltration membrane with a molecular weight cutoff of 30 kDa to separate the supernatant obtained from the pretreatment, and retaining the concentrated solution.

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