Method for preparing chondroitin sulfate from sturgeon cartilage

A technology of chondroitin sulfate and cartilage, which is applied in the field of seafood processing, can solve the problems of unfavorable industrial production of chondroitin sulfate, low purity of chondroitin sulfate, and complicated extraction process, so as to shorten purification time, shorten extraction time, and improve extraction The effect of acquisition rate

Inactive Publication Date: 2013-02-13

SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI

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AI-Extracted Technical Summary

Problems solved by technology

[0004] The traditional extraction method of chondroitin sulfate in animal cartilage tissue is time-consuming and complicated, and the pur...

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Abstract

The invention discloses a method for preparing chondroitin sulfate from sturgeon cartilage. The method comprises the following steps: (1) pretreating cartilage; (2) performing ultrasonic enzymolysis treatment: hydrolyzing the sturgeon cartilage by using trypsin, accelerating the enzymolysis process under the action of ultrasonic waves, finishing enzymolysis, deactivating enzyme, cooling and centrifuging to obtain supernate containing sturgeon chondroitin; (3) hydrolyzing and precipitating synchronously, continuously hydrolyzing the supernate in the step (2) with ethanol and NaOH solution and centrifuging to obtain white precipitate; and (4) purifying, drying and redissolving the precipitate to remove insoluble impurities, and drying to obtain a white spongy chondroitin sulfate product. The process is simple to operate and is new. Based on the steps of performing ultrasonic auxiliary enzymolysis, hydrolyzing and precipitating synchronously and purifying, the hydrolysis time can be greatly shortened, the purity and the yield of the products are increased and the production cost is reduced.

Technology Topic

Ultrasonic assistedChondroitin +6

Examples

Experimental program(3)

Example Embodiment

[0012] Example 1

[0013] The method for extracting and purifying sturgeon chondroitin sulfate provided in this embodiment includes the following steps: 1) Sample pretreatment: take the sturgeon cartilage tissue and treat it in a water bath at 80°C for 20 minutes to remove impurities such as muscle and fat on the surface. Wet ultramicro Crush to make a wet sample with a particle size of 100 mesh.

[0014] 2) Extraction of sturgeon sulphuric acid cartilage: wet cartilage sample with 0.1M phosphate buffer solution, pH 6.5 mass volume ratio of 10 (g/ml), the enzyme used for enzymatic hydrolysis is trypsin, and the added amount is per gram of sturgeon The fish cartilage sample is added to 1000U. The ultrasonic-assisted enzymatic hydrolysis conditions were 35kHz frequency, 280W power, and 50°C temperature adjustment for 50min, then the enzyme was inactivated at 95°C for 5min, and the supernatant was obtained by freezing and centrifuging at 4°C for 10min.

[0015] 3) Hydrolysis and purification of sturgeon cartilage with sulfate: the volume of absolute ethanol is 0.8 times (V/V) of the supernatant, and the concentration of NaOH is 0.8M. NaOH was added to the absolute ethanol solution, mixed and pre-cooled, and then slowly added to the supernatant obtained by enzymatic hydrolysis. The hydrolysis and precipitation temperature was normal temperature and the time was 30 minutes. The precipitate is reconstituted with 2 times water, neutralized to neutral with 40% acetic acid by mass percentage, centrifuged at 4°C, and freeze-dried in vacuo to obtain a white spongy chondroitin sulfate product. The product yield is 28%, and the purity exceeds 79.2%.

Example Embodiment

[0016] Example 2

[0017] The method for extracting and purifying sturgeon chondroitin sulfate provided in this embodiment includes the following steps: 1) Sample pretreatment: take the sturgeon cartilage tissue and treat it in a water bath at 90°C for 10 minutes to remove impurities such as muscle and fat on the surface, dry, crush, and dry The temperature is 50°C, and it is crushed to make a dry sample with a particle size of 80 mesh.

[0018] 2) Extraction of sturgeon sulfate cartilage: dry cartilage sample with 0.1M phosphate buffer solution, pH 7.0 mass volume ratio of 10 (g/ml), the enzyme used for enzymatic hydrolysis is trypsin, and the amount added per gram of sturgeon The cartilage sample was added to 2000U. The ultrasonic-assisted enzymatic hydrolysis conditions are 35kHz frequency, 360W power, and temperature adjusted to 40°C for enzymatic hydrolysis for 20 minutes, then the enzyme is inactivated at 95°C for 5 minutes, and the supernatant is obtained by freezing and centrifuging at 3°C for 10 minutes.

[0019] 3) Purification of sturgeon sulfate cartilage: The volume of absolute ethanol used for the hydrolysis and precipitation is 1.2 times (V/V) of the supernatant, and the concentration of NaOH is 0.6M. The specific steps of hydrolysis and precipitation are adding NaOH solution to a quantitative absolute ethanol solution, mixing and pre-cooling, and slowly adding it to the supernatant obtained from enzymatic hydrolysis. The hydrolysis and precipitation temperature is normal temperature and the time is 35 minutes. The precipitate was reconstituted with 3 times water, neutralized with 40% acetic acid to neutrality, centrifuged at 2°C, and vacuum freeze-dried to obtain a white spongy chondroitin sulfate product. The product yield is 29.61% and the purity is 76.3%.

Example Embodiment

[0020] Example 3

[0021] The method for extracting and purifying sturgeon chondroitin sulfate provided in this embodiment includes the following steps:

[0022] 1) Sample pretreatment: Take the sturgeon cartilage tissue and treat it in a water bath at 85°C for 20 minutes to remove impurities such as muscle and fat on the surface. The drying temperature is 48°C and the crushed particle size is 120 mesh.

[0023] 2) Sturgeon sulfate cartilage extraction: the mass-volume ratio of cartilage sample to phosphate buffer solution 0.1M, pH 7.0 is 10 (g/ml), the enzyme used for enzymatic hydrolysis is trypsin, and the added amount is per gram of sturgeon cartilage The sample is added to 2000U. The ultrasound-assisted enzymatic hydrolysis conditions were 35kHz frequency, 360W power, and temperature adjusted to 45°C for enzymatic hydrolysis for 30 minutes, then the enzyme was inactivated at 95°C for 5 minutes, and the supernatant was obtained by freezing and centrifuging at 0°C for 10 minutes.

[0024] 3) Purification of sturgeon sulfate cartilage: The volume of absolute ethanol used for the hydrolysis and precipitation is 1.0 times (V/V) of the supernatant, and the NaOH concentration is 0.8M. The specific steps of hydrolysis and precipitation are adding NaOH to the absolute ethanol solution, mixing and pre-cooling, and then slowly adding it to the supernatant obtained from enzymatic hydrolysis. The hydrolysis and precipitation temperature is normal temperature and the time is 40 minutes. The precipitate was re-dissolved with 3 times water, neutralized with 40% acetic acid, centrifuged at 4°C, and freeze-dried in vacuo to obtain a white spongy chondroitin sulfate product. The product yield is 31.0% and the purity is 81.3%.

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Classification and recommendation of technical efficacy words

Reduce purification time
Reduce extraction time

Method for preparing chondroitin sulfate from sturgeon cartilage

AI-Extracted Technical Summary

Problems solved by technology

Abstract

Examples

Example Embodiment

Example Embodiment

Example Embodiment

PUM

Description & Claims & Application Information

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