CN102584611B - Production method for medical grade valine - Google Patents

Abstract

The invention discloses a production method for high purity medical grade valine. Industrial valine raw material is prepared to be water solution higher than 5 percent under the temperature of 50 to 65 DEG C, flocculating agent is added, modified activated carbon is added after filtering, ultrafiltration is performed by using an ultra filtration membrane with molecular weight cutoff larger than 500, then vacuum concentration and cooling crystallization are performed to obtain the crystal product of medical grade valine, mother solution is added into a gel chromatography column for chromatographic separation, pure water with the temperature of 40 to 50 DEG C is taken as eluate, valine solution flowing out of the column is collected, after concentrated crystallization, the crystal product of medical grade valine is obtained, and finally, the removing rate of bacterial endotoxin and protein in the product can reach 99 percent, lactamic acid, aminocarproic acid and isoleucine can be effectively removed, the purity of the product can reach 99 percent, medical grade standards are met, the comprehensive yield of products can reach more than 96 percent, and large-scale production can be achieved.

Description

A kind of production method of medical grade valine

Technical field

The present invention relates to a kind of amino acid whose separation and method of purification, especially the production method of high purity medical level Valine.

Background technology

Valine belongs to branched chain amino acid, is one of necessary amino acid of human body, has different physiological roles, is widely used in the fields such as manufacture of medicine, food and seasonings, animal-feed and makeup.At field of medicaments, α-amino-isovaleric acid is usually used in manufacturing aminoacids complex transfusion and amino acids oral-liquor hardens and renal failure with treatment hemato encephalic barrier, hepatic coma, Chronic Liver, inborn errors of metabolism, septicemia and diabetes etc., also can be used as the medicine of accelerating surgical wound healing.

The main production method of α-amino-isovaleric acid is fermentation method, usings glucose as nutritive substance, after corresponding enzymic fermentation, produces α-amino-isovaleric acid.During the fermentation, because the raw material sugar adding can not all be utilized, valine fermentation liquid is also carried the material that polysaccharide, non-reducing sugar etc. can not be fermented utilization secretly, therefore when fermentation ends, in fermented liquid except existing α-amino-isovaleric acid, also have protein, pigment, colloid thing, raw material reducing sugar, polysaccharide, non-reducing sugar etc., and amino acid heteroacid is as leucine, L-Ala, Isoleucine.

The gordian technique of purifying valine is the separation of heteroacid in α-amino-isovaleric acid, and the common method of existing purifying valine has recrystallization method, the precipitator method and ion-exchange techniques.

The method of recrystallization is to utilize gac to carry out pre-treatment to α-amino-isovaleric acid, and the liquid after decolouring is through heating, vacuum concentration, crystallisation by cooling.The mother liquor of separating is got back to down in the filtrate of criticizing and recycle after treatment, although this method is simple to operate, but the separated heteroacid of main difficulty (L-Ala, leucine) existing due to α-amino-isovaleric acid is neutral amino acids, therefore, at mother liquid recycle, arrive after certain degree, amino acid heteroacid just can not effectively be removed, and a large amount of mother liquors cannot utilize, and reduces the yield of α-amino-isovaleric acid.

Ajincomoto Co., Inc adopts p-isopropyl Phenylsulfonic acid to precipitate purifying valine, although this intermediate processing is simple to operate, selectivity is strong, but precipitation agent reclaims difficulty, discharging of waste liquid pollutes, the toxicity of residual precipitation agent is larger, and the amino acid of food grade and pharmaceutical grade can not adopt this method to produce.

What the extraction of food grade and medical grade valine generally adopted is ion exchange method, but this method exists following problem: the one, adopt the method for filtering remove thalline, because thalline is smaller, thalline is removed not thorough conventionally; The 2nd, when depigmentation, bacterial endotoxin impurity, to use large carbon content active, and need to repeatedly process, technique relative complex, pollutes greatlyr, and simultaneously gac also adsorbs the more α-amino-isovaleric acid of taking, and yield is reduced.The 3rd, in α-amino-isovaleric acid, impurity component is many, and content is high, and the 4th, resin stain is serious, easily cause the destruction of resin structure and lose regenerative power, increase running cost, the high ionic substance of content can make resin saturated fast simultaneously, the regeneration times that increases resin, causes the problems such as sewage discharge.

CN101721979A discloses a kind of macroporous adsorbent resin special for separating valine, use this resin can α-amino-isovaleric acid, L-Ala, leucine is separated, product purity can reach more than 99%, but the production of this resin is still at the experimental stage, can not large-scale application in industrial production.

In addition, Chinese patent application prospectus CN101798273A adopts and is warmed to 220 ± 10 ℃ of degree, and L-Ala and Isoleucine are removed in distillation, but under high temperature, α-amino-isovaleric acid easily decomposes destruction.

Summary of the invention

The object of this invention is to provide a kind of industrial scale and produce the method for medical grade valine, the method can be removed the heteroacid in α-amino-isovaleric acid effectively, improves α-amino-isovaleric acid yield, reduces production costs.

The inventive method comprises the steps:

A. industrial α-amino-isovaleric acid raw material is mixed with to more than 5% aqueous solution of weight percent concentration at 50 ~ 65 ℃, adds the flocculation agent of solution weight 0.2 ~ 2%, stir after 1 ~ 5 hour and filter;

B. in filtrate, add the modified activated carbon of filtrate weight 0.5 ~ 2.5%, 50 ~ 65 ℃ of stirrings, after 1 ~ 5 hour, filter;

C. filtrate, at 50 ~ 65 ℃, is greater than 500 ultra-filtration membrane ultrafiltration with molecular weight cut-off;

D. the filtrate vacuum concentration after ultrafiltration, crystallisation by cooling, filters, and obtains medical grade valine crystalline product.

In the present invention, the industrial α-amino-isovaleric acid aqueous solution is preferably mixed with the saturated aqueous solution under above-mentioned solvent temperature.

The present invention's flocculation agent used can be by natural polymer substance as the polymkeric substance with acrylic amide graft copolymerization such as starch, Mierocrystalline cellulose, chitosan, or polyacrylamide.Preferably polyacrylamide, can remove microorganism, bacterial endotoxin and fragment in α-amino-isovaleric acid mother liquor effectively, and part that can be removed pigment, heavy metal ion and some other pollutent.The add-on of flocculation agent preferably solution weight 0.5 ~ 1.5%.

Modified activated carbon is that gac is immersed in to pH=4 ~ 6.5, and then the aqueous hydrogen peroxide solution of weight concentration 20 ~ 50% filters, and washing is dry, then the product obtaining after activation at 160 ~ 250 ℃.The add-on of modified activated carbon preferably solution weight 1 ~ 2%.The present invention carries out oxide treatment to gac, can form on the surface of gac the oxidation carbon-coating of one deck band portion electric charge, adopt modified activated carbon as sorbing material, except having the decolouring function of gac, also there is macromole in adsorbent solution and comprise bacterial endotoxin, and the effect of zwitterion.

In ultra-filtration membrane ultra-filtration process, entering film pressure and membrane pressure difference can be 0.02 ~ 0.1MPa, preferably 0.06 ~ 0.08MPa.

α-amino-isovaleric acid solution after ultrafiltration can be at normal temperature to 60 ℃, concentrated under the air pressure of working pressure 0.05 ~ 0.09MPa.Then concentrated solution is cooled to 20 ~ 25 ℃ of crystallizations, filters, obtain medical grade valine crystalline product.

In a preferred method of the invention, also comprise step e: the mother liquor in step D after crystallization filtration joins gel chromatography column and carries out chromatographic separation, with the pure water of 40 ~ 50 ℃, as elutriant, collect the α-amino-isovaleric acid solution flowing out in post, through condensing crystal, obtain medical grade valine crystalline product.

More preferably: the filtrate vacuum Concentrating Process in step D after ultrafiltration stops concentrating after can being concentrated into original solution volume 20 ~ 40%, crystallisation by cooling filters out after crystalline product, and mother liquor joins gel chromatography column chromatography for separation.

In chromatographic separation, the velocity of flow of elutriant in gel column is 0.5 ~ 5cm/min, is preferably 1.5 ~ 3 cm/min.

Above-mentioned gel is dextrane gel or polyacrylamide gel, preferably polyacrylamide gel.

The present invention takes the absorption of flocculation agent-modified activated carbon, the production method of crystallization and gel chromatography technical tie-up, muriate in the finished product, vitriol, ammonium salt plasma reaches version < < Chinese Pharmacopoeia > > in 2010 requirement, bacterial endotoxin, protein removal rate reaches more than 99%, pigment removal reaches 99%, effectively by L-Ala, leucine, Isoleucine separation is removed, the purity of product reaches more than 99%, reach pharmaceutical grade standard, the comprehensive yield of product reaches more than 96%, be produced on a large scale, equipment is simple, easy and simple to handle, gel is recyclable to be used again.

Accompanying drawing explanation

Fig. 1 is technological process of production sketch of the present invention.

Embodiment

The preparation of modified activated carbon: under normal temperature, gac is immersed in to PH=6, the aqueous hydrogen peroxide solution that weight concentration is 30%, after 10 hours, filters, with pure water, wash to washings without acid ion, dry, then at 200 ℃, dry activation 6 hours, obtain modified activated carbon.

In dissolving vessel, pump enters pure water 1000L, be heated to 60 ℃, add industrial α-amino-isovaleric acid raw material 100.40Kg, unlatching is stirred to dissolve complete, be incubated about 6 hours, (Shanghai Mei Naiqing Trade Co., Ltd. produces to add 10.02Kg polyacrylamide flocculant, model is the polyacrylamide of Kingfloc4170), stir 3 hours, remove by filter the protein of flocculation agent and absorption thereof, microorganism and fragment, filter, in filtrate, add 14.50Kg modified activated carbon, stir and decolour and remove bacterial endotoxin for 3 hours, filter, filtrate is at 58 ~ 62 ℃, enter under the condition that film pressure and membrane pressure difference are 0.07MPa with holding back relative molecular weight and be greater than 500 ultra-filtration membrane and carry out heat filtering.

Start vacuum pump, the filtrate after ultrafiltration, at 60 ℃, stops concentrating under the pressure lower than 0.09MPa for 1/3 o'clock that is concentrated into original solution volume, and regulating pH value is 6.0, is cooled to 25 ℃ of crystallizations, and suction filtration obtains α-amino-isovaleric acid crystalline product 61.24Kg.

Gel chromatography method purifying valine for mother liquor, filtrate is added to 1.5 meters of high Polyacrylamide Gel Columns, and (Switzerland Pharmacia Biotech company produces, model is the polyacrylamide gel of sephadexG100), adopt the pure water of 40 ℃ as elutriant, controlling the velocity of flow of elutriant in gel column is 2cm/min, collect the elutriant first flowing out in post, use differential refraction detector on-line monitoring, when there is leucine, stop collecting, elutriant is at 60 ℃, concentrated under 0.08Mpa working pressure, crystallization while being then cooled to 25 ℃, with centrifugal concentrator, throw away mother liquor, rotary-drum vacuum is dry, obtain α-amino-isovaleric acid crystalline product 31.33Kg.

Accumulative total obtains α-amino-isovaleric acid crystalline product 92.57Kg, total recovery 96.04%.

Primary industry α-amino-isovaleric acid part index number: muriate>=0.05%, vitriol>=0.05%, ammonium salt>=0.05%, transmittance 81%(C=10, H₂o), amino acid heteroacid: 1.2 ~ 1.5%, bacterial endotoxin≤1000EU/mg, valine content 96%.

Crystalline product α-amino-isovaleric acid part index number: muriate≤0.02%, vitriol≤0.03%, ammonium salt≤0.02%, transmittance 99.1%(C=10, H₂o), amino acid heteroacid < 0.2%, bacterial endotoxin < 10EU/g, valine content 99.3%, reaches the standard of medical grade valine.

Mother liquor crystallization product α-amino-isovaleric acid part index number: muriate≤0.02%, vitriol≤0.03%, ammonium salt≤0.02%, transmittance 99.0%(C=10, H₂o), amino acid heteroacid < 0.5%, bacterial endotoxin < 20EU/g, valine content 99.0%, reaches the standard of medical grade valine.

Claims (12)

1. a production method for medical grade valine, is characterized in that, comprises the steps:

A. industrial α-amino-isovaleric acid raw material is mixed with to more than 5% aqueous solution of weight percent concentration at 50 ~ 65 ℃, adds the flocculation agent of solution weight 0.2 ~ 2%, stir after 1 ~ 5 hour and filter;

B. in filtrate, add the modified activated carbon of filtrate weight 0.5 ~ 2.5%, 50 ~ 65 ℃ of stirrings, after 1 ~ 5 hour, filter;

C. filtrate, at 50 ~ 65 ℃, is greater than 500 ultra-filtration membrane ultrafiltration with molecular weight cut-off;

D. the filtrate vacuum concentration after ultrafiltration, crystallisation by cooling, filters, and obtains medical grade valine crystalline product;

Described modified activated carbon is that gac is immersed in to pH=4 ~ 6.5, and then the aqueous hydrogen peroxide solution of weight concentration 20 ~ 50% filters, and washing is dry, then the product obtaining after activation at 160 ~ 250 ℃.

2. the production method of medical grade valine according to claim 1, is characterized in that, in steps A, industrial α-amino-isovaleric acid is mixed with the saturated aqueous solution under described solvent temperature.

3. the production method of medical grade valine according to claim 1, is characterized in that, described flocculation agent is polyacrylamide.

4. the production method of medical grade valine according to claim 1, is characterized in that, in step C, the difference that solution flows to the pressure of ultra-filtration membrane and flows out the pressure of ultra-filtration membrane is 0.02 ~ 0.1MPa.

5. the production method of medical grade valine according to claim 4, is characterized in that, in step C, the difference that solution flows to the pressure of ultra-filtration membrane and flows out the pressure of ultra-filtration membrane is 0.06 ~ 0.08MPa.

6. the production method of medical grade valine according to claim 1, is characterized in that, the add-on of described modified activated carbon is 1 ~ 2% of solution weight.

7. according to the production method of medical grade valine described in the arbitrary claim of claim 1 ~ 6, it is characterized in that, also comprise step e: the mother liquor in step D after crystallization filtration joins gel chromatography column and carries out chromatographic separation, with the pure water of 40 ~ 50 ℃ as elutriant, collect the α-amino-isovaleric acid solution flowing out in post, through condensing crystal, obtain medical grade valine crystalline product.

8. the production method of medical grade valine according to claim 7, is characterized in that, in step D, filtrate vacuum concentration is to original solution volume 20 ~ 40%.

9. the production method of medical grade valine according to claim 7, is characterized in that, in described chromatographic separation, the velocity of flow of elutriant in gel column is 0.5 ~ 5cm/min.

10. the production method of medical grade valine according to claim 8, is characterized in that, in described chromatographic separation, the velocity of flow of elutriant in gel column is 0.5 ~ 5cm/min.

11. production methods of medical grade valine according to claim 7, is characterized in that, described gel is polyacrylamide gel.

12. production methods of medical grade valine according to claim 8, is characterized in that, described gel is polyacrylamide gel.

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