Method for large-scale preparation of high-purity unsaturated hyaluronic acid disaccharide
A technology for large-scale preparation and hyaluronic acid, which is applied in the field of oligomeric hyaluronic acid preparation, can solve problems such as unfavorable industrial production, low product purity and reduction, and achieve the effects of less impurities, high product purity and good precipitation effect.
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Embodiment 1
[0046] 1. To 1.5 m 3 Add 1 m to the stainless steel dissolution tank 3 Purify water, raise the temperature to 30°C, and adjust the pH to 5.0 with hydrochloric acid.
[0047] 2. Set the stirring speed to 20 rpm, and add 1.8×10 8 IU of Bacillus hyaluronidase, after stirring for 1 min, slowly add 100 kg of sodium hyaluronate with a molecular weight of 2000 kDa for 1 h. Add 50 kg of sodium hyaluronate for 1 hour, continue enzymatic hydrolysis for 1 hour after adding, then continue to slowly add 30 kg of sodium hyaluronate with a molecular weight of 2000 kDa for 1 hour, continue enzymatic hydrolysis until the molecular weight is reached When it is less than 50 kDa, raise the temperature to 50°C and maintain it for 60 min.
[0048] 3. Filter the enzymatic hydrolyzate after deactivation with a 0.45 µm mixed cellulose filter, and then filter it with a 3 m 3 Precipitate with ethanol to obtain sodium hyaluronate precipitate, dehydrate the precipitate with ethanol, and then vacuum dr...
Embodiment 2
[0052] 1. To 1.5 m 3 Add 1 m to the stainless steel dissolution tank 3 Purify water, raise the temperature to 35°C, and adjust the pH to 6.0 with hydrochloric acid.
[0053] 2. Set the stirring speed to 30 rpm, and add 2.4×10 7 IU of Bacillus hyaluronidase, after stirring for 1 min, slowly add 80 kg of potassium hyaluronate with a molecular weight of 1300 kDa for 1 h, enzymatic hydrolysis for 1 h after adding, and then continue to slowly add potassium hyaluronate with a molecular weight of 1300 kDa Add 40 kg of potassium hyaluronate for 1 hour, continue enzymatic hydrolysis for 1 hour after adding, then continue to slowly add 20 kg of potassium hyaluronate with a molecular weight of 1300 kDa for 1 hour, continue enzymatic hydrolysis until When the molecular weight is less than 80 kDa, raise the temperature to 55°C and maintain it for 40 min.
[0054] 3. Filter the enzymatic hydrolyzate after deactivation with a 0.45 µm mixed cellulose filter, and then filter it with a 5 m ...
Embodiment 3
[0058] 1. To 1.5 m 3 Add 1 m to the stainless steel dissolution tank 3 Purify water, raise the temperature to 40°C, and adjust the pH to 6.5 with hydrochloric acid.
[0059] 2. Set the stirring speed to 50 rpm, and add 1.0×10 7 IU of Bacillus hyaluronidase, after stirring for 1 min, slowly add 40 kg of calcium hyaluronate with a molecular weight of 1800 kDa for 1 h, enzymatic hydrolysis for 1 h after adding, and then continue to slowly add calcium hyaluronate with a molecular weight of 1800 kDa Add 35 kg of calcium hyaluronate for 1 hour, continue enzymatic hydrolysis for 1 hour after adding, then continue to slowly add 25 kg of calcium hyaluronate with a molecular weight of 1800 kDa for 1 hour, continue enzymatic hydrolysis until the molecular weight is reached When it is less than 55 kDa, raise the temperature to 60°C and maintain it for 20 min.
[0060] 3. Filter the enzymatic hydrolysis solution after deactivation with a 0.45 µm mixed cellulose filter, and then filter i...
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