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Purification method of nicotinamide adenine dinucleotide

A technology of nicotinamide adenine and dinucleotide, which is applied in chemical instruments and methods, organic chemistry, preparation of sugar derivatives, etc., can solve the problems of cumbersome process, low product quality, complicated operation process, etc., and achieve process operation. The effect of simplicity, good product quality and low process cost

Pending Publication Date: 2022-04-01
JIANGSU CHENGXIN PHARMA +1
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Problems solved by technology

[0005] CN104876993A discloses a method for separating and purifying NAD+ using a reverse HPLC preparative column, but there are problems of high cost and difficulty in industrialization
[0006] CN111065644A discloses a method for purifying NAD+ by using crystallization to increase temperature and filter multiple times to adjust pH; however, the method is complicated to operate, and there is a risk of product decomposition during the process of pH adjustment, and the crystallization process uses Chemical reagent ethanol, organic reagents will remain in the product
However, multiple purifications in this process (D152 resin, 717 resin, modified silica gel preparation chromatography), multiple nanofiltration concentration, and complicated process, especially when purified by D152, ammonia water is used for elution, and the product is under alkaline conditions. Unstable, there is a risk of decomposition; in the pure product preparation process, the use of modified silica gel to prepare chromatographic separation, the cost is high, the organic solvent acetone crystallization is used in the crude product preparation process, and the final product quality is not high (the highest purity is 99.3%)
[0009] CN107417749A discloses a resin filler separation method for coenzyme I. The filtered cell filtrate containing coenzyme I is washed and eluted after exchanged by a D945 macroporous resin column. The D945 macroporous resin has a large specific surface area, strong adsorption, High selectivity, high reuse rate, no need to adjust PH, relatively mild conditions, and can be separated in the most suitable pH range of coenzyme I, reducing the generation rate of by-products, greatly improving the coenzyme I after resin separation Purity; however, the macroporous resin requires organic solvent ethanol in the elution and regeneration process, and the cost is high. Due to the presence of organic solvent in the eluent, there are certain problems in the subsequent removal process; the method of nanofiltration concentration is used to remove ethanol, Organic solvents will damage ordinary nanofiltration membranes; removing ethanol by heating will damage the product

Method used

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  • Purification method of nicotinamide adenine dinucleotide
  • Purification method of nicotinamide adenine dinucleotide
  • Purification method of nicotinamide adenine dinucleotide

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Embodiment 1

[0091] This embodiment provides a method for purifying nicotinamide adenine dinucleotide, which includes the following steps:

[0092] (1) Take 2.5kg of reaction solution (wherein the effective amount of NAD+ is 50g), first filter through medium-speed filter paper to remove insoluble protein, and obtain supernatant;

[0093] (2) The supernatant that step (1) is obtained is carried out ultrafiltration processing by vacuum fiber membrane (molecular weight cut-off 8000-10000D); After ultrafiltration finishes, add 1L purified water in the liquid that ultrafiltration obtains and continue ultrafiltration (repeat Twice operation), obtain ultrafiltrate;

[0094] (3) carry out anion resin purification (D301 type, Cl type) purification to the ultrafiltrate that step (2) obtains; First to diameter, be 10cm, add 2L anion exchange resin in the chromatographic column that is 50cm high; Then step ( 2) The obtained ultrafiltrate is loaded at a flow rate of 1 column volume / h; after loading, r...

Embodiment 2

[0101] This embodiment provides a method for purifying nicotinamide adenine dinucleotide, which includes the following steps:

[0102] (1) Take 2.5kg of reaction solution (wherein the effective amount of NAD+ is 50g), first filter through medium-speed filter paper to remove insoluble protein, and obtain supernatant;

[0103] (2) The supernatant that step (1) is obtained is carried out ultrafiltration processing by vacuum fiber membrane (molecular weight cut-off 8000-10000D); After ultrafiltration finishes, add 1.1L purified water in the liquid that ultrafiltration obtains and continue ultrafiltration ( Repeat twice) to obtain ultrafiltrate;

[0104] (3) carry out anion resin purification (D301 type, Cl type) purification to the ultrafiltrate that step (2) obtains; First to diameter, be 10cm, add 2.2L anion exchange resin in the chromatographic column that is 50cm high; Then step (2) The obtained ultrafiltrate is loaded at a flow rate of 1.2 times the column volume / h; after th...

Embodiment 3

[0109] This embodiment provides a method for purifying nicotinamide adenine dinucleotide, which includes the following steps:

[0110] (1) Take 2.5kg of reaction solution (wherein the effective amount of NAD+ is 50g), first filter through medium-speed filter paper to remove insoluble protein, and obtain supernatant;

[0111] (2) the supernatant that step (1) is obtained is carried out ultrafiltration processing by vacuum fiber membrane (molecular weight cut-off 8000-10000D); After ultrafiltration finishes, add 0.9L purified water in the liquid that ultrafiltration obtains and continue ultrafiltration ( Repeat twice) to obtain ultrafiltrate;

[0112] (3) carry out anion resin purification (D301 type, Cl type) purification to the ultrafiltrate that step (2) obtains; First to diameter, be 10cm, add 1.9L anion exchange resin in the chromatographic column that is 50cm high; Then step (2) The obtained ultrafiltrate was loaded at a flow rate of 1.1 times the column volume / h; after t...

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Abstract

The invention provides a purification method of nicotinamide adenine dinucleotide, which comprises the following steps: sequentially carrying out solid-liquid separation, ultrafiltration, anion exchange resin treatment, nanofiltration, decoloration and drying on reaction liquid to obtain high-purity nicotinamide adenine dinucleotide. According to the purification method of nicotinamide adenine dinucleotide provided by the invention, purified water is adopted in anion exchange resin flushing and elution processes, and an organic solvent is not needed for flushing and elution, so that the purification method is more environment-friendly and lower in cost; the eluent only needs to be subjected to vacuum freeze drying after being concentrated, and compared with a crystallization mode, the product yield is high, no chemical reagent residues exist in the product, and the product quality is better; the purification method of nicotinamide adenine dinucleotide provided by the invention has the remarkable advantages of simple process operation, high yield, good product quality, environmental protection and low cost.

Description

technical field [0001] The invention belongs to the technical field of nucleotide coenzymes, and in particular relates to a method for purifying nicotinamide adenine dinucleotide. Background technique [0002] Nicotinamide adenine dinucleotide (NAD+) is an essential multifunctional small molecule for organisms, mainly serving as an important coenzyme for oxidoreductases and a source of NADP (nicotinamide adenine dinucleotide phosphate). The multi-enzyme redox system in which NAD(H) participates is the main biological oxidation system in the electron transfer process in the respiratory chain of living organisms. Most of the oxidation reactions in the decomposition of the three major metabolites of sugar, lipid and protein are also through this system. The system is completed. NAD+ has great application value in the field of medicine. [0003] The preparation methods of nicotinamide adenine dinucleotide (NAD+) are mainly divided into chemical synthesis and biocatalysis. Chem...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H19/207C07H1/06
Inventor 李斌李二军魏云亮张超祝俊徐飞谭从祥何宝亮张建明
Owner JIANGSU CHENGXIN PHARMA
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