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Method for purifying citicoline from biotransformation or multienzyme reaction liquid

A technology of citicoline and enzyme reaction solution, which is applied in the field of separation and purification of citicoline, can solve the problem of cytidine acid being unable to be recovered, and achieves the effects of excellent product quality, good separation effect and easy operation.

Active Publication Date: 2008-01-02
NANTONG QIUZHIYOU BIOSCI & BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the more expensive unreacted cytidylic acid cannot be recovered

Method used

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  • Method for purifying citicoline from biotransformation or multienzyme reaction liquid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] 4000mL LCDP-C reaction liquid, containing CDP-C 34.5g, CMP 5.0g, phosphate 150g, phosphorylcholine 40g, magnesium sulfate 9.7g, protein 107mg, pass 1-3 in order #Column, the flow rate is 0.3m / h. The size of column 1 is 40×1000mm, containing 1000mL of treated HD-1(H+) resin; the size of column 2 is 60×1000mm, containing 2000mL of treated D315(OH-) resin; the size of column 3 is 30×1000mm, Pack 500mL of treated HZ201 resin.

[0038] After loading the column, rinse with 1000mL deionized water. Disconnect column 1 and column 2. Then column 2 was rinsed with 1500 mL of dilute hydrochloric acid solution at pH 5.0. Disconnect column 2 and column 3. Column 3 was then eluted with a pH 2 dilute hydrochloric acid solution (containing 4% sodium chloride). Collect about 800 mL of the eluate containing CDP-C, concentrate to 100 mL, and adjust the pH to 7.5 with sodium hydroxide. Add 400mL of absolute ethanol, stir evenly, place at 4°C for 16 hours, filter and crystallize, and d...

Embodiment 2

[0041] 4000mL of the CDP-reaction solution in Example 1 was passed through 1-3 successively # Column, the flow rate is 0.3m / h.

[0042] The size of column 1 is 40×1000mm, containing 1000mL of treated HD-1(H+) resin; the size of column 2 is 60×1000mm, containing 2200mL of treated D301(OH-) resin; Pack 450 mL of treated Amberlite IRA-900 (OH-) resin.

[0043] Others are operated according to Example 1.

[0044] As a result, 28.5 g of CDP-C sodium salt was obtained with a purity of 98.9% and a yield of 82.6 g. 4.0 g of cytidylic acid was recovered with a yield of 80%.

Embodiment 3

[0046] The CDP-C (A) obtained in Example 1 and the CDP-C (B) prepared by the traditional process were prepared into a 0.125 g / mL solution with sterile water, vacuum-sealed, and heated at 110° C. for 30 minutes. After cooling, the absorbance was measured at 430nm, and the results are shown in Table 1.

[0047] Table 1

[0048] sample

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Abstract

The invention discloses a separating method of CDP-C from biological conversion or polyzyme reacting liquid in the biochemical technical domain, which is characterized by the following: passing the CDP-C reacting liquid into cation exchange column 1, anion exchange column 2 and anion exchange column 3; washing the columns 1-3 through deionized water; washing the anion exchange column 2 and anion exchange column 3 through solution with pH value at 5.0; eluting CDP-C from anion exchange column 3 through high-density salt and alcaine to obtain high-density CDP-C solution; using the solution with pH value at 3.0 to elute CMP on the anion exchange column 2. The invention shortens the separating time with water consumption over 60%, which improves the receiving rate of CDP-C with easy crystallization and recycles most of non-reacted CMP.

Description

technical field [0001] The invention relates to a method for separating and purifying citicoline. In particular, the method for separating and purifying citicoline from biotransformation or multi-enzyme reaction liquid by adopting novel ion exchange resin combined column chromatography. Background technique [0002] Citicoline diphosphate choline (CDP-Choline for short, CDP-C) is an important intermediate in the synthesis of lecithin in the body and a necessary coenzyme for phospholipid metabolism. Clinically, CDP-C can improve the metabolism of phospholipids in the state of impaired phospholipid metabolism in the brain, thereby promoting the recovery of consciousness. It has a certain curative effect on the disturbance of consciousness caused by traumatic brain injury and brain surgery, as well as some chronic diseases such as stroke sequelae, Parkinson's disease, central vertigo, tinnitus, glaucoma and sensory deafness. [0003] CDP-C can be prepared by chemical synthesi...

Claims

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Application Information

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IPC IPC(8): C07H19/10C07H1/06
Inventor 邱蔚然闫蓬勃丁庆豹
Owner NANTONG QIUZHIYOU BIOSCI & BIOTECH
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