CN108864271A - A kind of preparation method of iron protein succinylate - Google Patents
A kind of preparation method of iron protein succinylate Download PDFInfo
- Publication number
- CN108864271A CN108864271A CN201710330921.8A CN201710330921A CN108864271A CN 108864271 A CN108864271 A CN 108864271A CN 201710330921 A CN201710330921 A CN 201710330921A CN 108864271 A CN108864271 A CN 108864271A
- Authority
- CN
- China
- Prior art keywords
- added
- solution
- iron
- succinyl
- iron protein
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4732—Casein
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Gastroenterology & Hepatology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The present invention relates to pharmaceutical technology fields, specifically disclose a kind of preparation method of high acylation degree iron protein succinylate.On the basis of current iron protein succinylate preparation method, by preparing acylation degree height（Acylation degree > 99.3%）Succinyl albumen, finally prepare the high iron protein succinylate of acylation degree, increase its hydrophilic radical, increase its water-soluble, reduce its Aqueous Solution Viscosity, solve the problems, such as that iron protein succinylate oral solution large viscosity, sediment are more.
Description
Technical field
The present invention relates to pharmaceutical technology fields, and in particular to a kind of preparation method of high acylation degree iron protein succinylate.
Background technique
Iron and emulsion succinic acid protein binding in iron protein succinylate form iron-protein complex, various to treat
Hypoferric anemia.Its structure is as follows：
Iron protein succinylate is in precipitated form when pH value is less than 4, and when acidity-basicity ph is higher (pH7.5~8)
Again become solable matter.In addition, iron protein succinylate cannot be by pepsin digestion, it but can be by pancreas egg in pH neutral
White enzyme hydrolysis.Due to these properties of iron protein succinylate, contained by iron by protein film protection and the salt in different gastric juice
Acid and pepsin react, because of the mucosal lesion without caused by.Iron in iron protein succinylate is opened in duodenum
Begin to discharge, because the raising of normal ph is so that this compound becomes solvable again, and makes protein film by trypsase institute
Digestion, the iron released is very beneficial for the physiological absorption of body, but not will form too high absorption peak；In fact, it is in
A kind of existing constant absorption trend, progressivelyes reach the best steady state of absorption and storage at each position of body.
1987, Italy developed a kind of entitled iron protein succinylate (iron-protein in the general big pharmaceutical factory of mark
Succinylate, abbreviation ISP) nanometer polypeptide albuminoid mend iron drug, in Italy's successfully listing and in global more than 20 a states
Family includes the list marketings successively such as Spain, Portugal, Greece, Argentina, South Korea, China.Iron protein succinylate oral solution
Apply for that import has been approved by home by ITALFARMACO S.A. company of Spain, trade name (Chinese) Ferplex, trade name
(English) Ferplex.
Mainly using casein as raw material, with succinic anhydride acylation reaction life occurs for existing iron protein succinylate preparation route
At succinyl albumen, ferric ion occurs load iron with succinyl albumen again and reacts, and generates iron protein succinylate.Reaction equation is such as
Under：
US4493829 discloses a kind of preparation method of iron protein succinylate.Using milk powder as raw material, under alkaline condition
(pH7.5~8) and succinic anhydride carry out acylation reaction, and after completion of the reaction, reaction solution is centrifuged or filtering, and hydrochloric acid acid is added in filtrate
Changing (pH2.5~3) makes to precipitate, and is centrifuged or filters to take and precipitates to obtain succinyl albumen.Succinyl albumen is dispersed in water, in alkaline item
It is dissolved under part (pH7.5), liquor ferri trichloridi is added and carries out carrying iron reaction, filters after completion of the reaction, it is thick to obtain iron protein succinylate
Product.Iron protein succinylate crude product is dispersed in water, and is dissolved at alkaline condition (pH7.5), and filtering obtains solution.It can be through with lower section
Method is prepared into iron protein succinylate：1) acidified (pH2.5), filtering, vacuum drying.2) solution through dialysis remove sodium chloride after,
Freeze-drying or spray drying.It is 95% that the method, which prepares products therefrom acylation degree,.
CN102838667 discloses a kind of preparation method of iron protein succinylate.Using casein as raw material, under alkaline condition
(pH9.0~10.0) and succinic anhydride carry out acylation reaction, and after completion of the reaction, hydrochloric acid acidification is added in filtrate in centrifugal filtration
(pH1~4) make to precipitate, and centrifuging and taking precipitates to obtain succinyl albumen.Succinyl albumen is dispersed in water, alkaline condition (pH9.0~
10.0) it is dissolved under, is centrifuged, obtains succinyl protein solution.Liquor ferri trichloridi is added, while sodium hydroxide is added and keeps reaction solution
PH9.0~10.0 carry out carrying iron reaction, are centrifuged after completion of the reaction, obtain solution.Hydrochloric acid acidification (pH1~4), which is added, to be made to precipitate, and is centrifuged
Obtain filter cake.It is redispersed in water, is dissolved under alkaline condition (pH9.0~10.0), be acidified (pH1~4), be centrifuged to obtain filter cake.Again
It is dispersed in water, is dissolved under alkaline condition (pH9.0~10.0), is centrifuged, acquired solution is after 1.2 μm of ceramic membrane refined filtrations, acid
Change (pH1~4), be centrifuged to obtain filter cake, iron protein succinylate is obtained after freeze-drying.The method prepares products therefrom acylation degree
95.0%~96.8%.
In above-mentioned iron protein succinylate oral solution process for preparation, poor, dissolution that generally there are iron protein succinylate dissolubilities
Speed is slow, and prepares gained oral administration solution large viscosity, the problems such as sediment is more.Iron protein succinylate solution viscosity is big, can lead
It causes to be difficult to carry out by the operating process that micro porous filtration removes microorganism in oral administration solution process for preparation.
The percentage of amino and the initial free amine group of casein that casein is acylated in acylation reaction is referred to as being acylated
Degree.The higher succinyl albumen of acylation degree, water solubility is better, Aqueous Solution Viscosity is lower, it is stronger to carry iron ability, corresponding final system
Standby gained iron protein succinylate good water solubility, Aqueous Solution Viscosity are low.It, can on the basis of current iron protein succinylate preparation method
By improving the acylation degree of iron protein succinylate, increasing its hydrophilic radical, increase its water-soluble and reduce its aqueous solution
Viscosity, to solve slow solution rate in iron protein succinylate oral solution process for preparation, oral administration solution large viscosity and sediment
More problems.
Summary of the invention
For the deficiencies in the prior art, the object of the present invention is to provide a kind of high acylation degree (acylation degrees>
99%), the iron protein succinylate that water-soluble is good, Aqueous Solution Viscosity is low, filter membrane passability is good, impurity content is low and quality is stable
Preparation method, described method includes following steps：
(1), in reaction vessel be added casein and 10~15 times of casein weight water, be added with stirring 1mol/L~
The pH to 7.0~10.0 that the sodium hydroxide solution of 4mol/L adjusts reaction solution dissolves casein.More batches are slowly divided to be added always again
Weight and casein weight ratio are 1:0.2~1:0.4 succinic anhydride, be added while adding succinic anhydride 1mol/L~
After the sodium hydroxide solution of 4mol/L concentration keeps reaction solution pH=7.0~10.0, succinic anhydride to be added, it is anti-to continue stirring
Answer 30~45min, then into reaction solution be added 1mol/L~5mol/L hydrochloric acid adjust pH=2~3, continue stirring 30~
45min, filtering, the pH=7 that obtained solid is washed to eluate, obtains succinyl albumen wet solid.
(2), the water dispersion of succinyl albumen wet solid 5~7 times of weight of casein inventory obtained by step (1), stirring
The lower sodium hydroxide solution that 1mol/L~4mol/L is added, which adjusts reaction solution pH=4.1~4.8, makes succinyl protein dissolution.Centrifugation
Or after filtering (aperture of filter material is 120~3 μm), obtain succinyl protein solution.
(3), the ferric chloride aqueous solutions of 1wt%~10wt% are slowly added to molten to succinyl albumen obtained by step (2)
In liquid, the weight ratio of succinyl albumen and ferro element is 1:0.45~1:0.55, after addition, continue 30~45min of stirring,
Filtering, the pH=7 that gained filter cake is washed to eluate, obtain iron protein succinylate crude product wet solid.
(4), the moisture of iron protein succinylate crude product wet solid 10~15 times of weight of casein inventory obtained by step (3)
It dissipates, the sodium hydroxide solution for being added with stirring 1mol/L~4mol/L, which adjusts reaction solution pH=7.0~10.0, makes albumen succinic acid
The dissolution of iron crude product.After 5 μm of filter membrane initial filters, then through 1 μm of filter membrane refined filtration, the hydrochloric acid of 1mol/L~5mol/L is added in acquired solution
PH=2~3, filtering are adjusted, obtained solid is washed to the pH=7 of eluate, obtains iron protein succinylate wet product.
(5), 10~50 mesh of oscillating granulator of iron protein succinylate wet product obtained by step (4) is pelletized, then sets boiling and does
Dry machine it is dry iron protein succinylate.
Further, the sodium hydroxide solution of addition 1mol/L~4mol/L described in step 2) adjusts reaction solution pH=4.1.
Further, 5 times of weight water dispersions of casein inventory of filter cake described in step 2).
Further, succinic anhydride total weight and casein weight ratio are 3:1.
Further, succinic anhydride is divided in 15-25 addition casein solution in step (1).
Further, actual conditions dry in step (5) are：First dry 0.5~1h at 20~30 DEG C, then it is warming up to 50
Dry 2~3h after~70 DEG C.
Further, actual conditions dry in step (5) are：The first dry 0.5h at 30 DEG C, then after being warming up to 70 DEG C
Dry 2h.
The present invention provides a kind of preparation method of iron protein succinylate, using casein as raw material, after succinic anhydride acylation
It is prepared into succinyl albumen, then is reacted with ferric iron complexing generation load iron and is prepared into iron protein succinylate crude product, most afterwards through refining
It is prepared into iron protein succinylate.Present invention preparation gained succinyl albumen acylation degree is higher than 99%, good water solubility, Aqueous Solution Viscosity
Low, filter membrane passability is good, impurity content is low, high income.
The present invention prepares the characteristic of gained iron protein succinylate product, greatly compensates for existing method and prepares albumen amber
Solution rate is slow, solution viscosity is high in following protein ferric succinate oral administration solution configuration process, solution sediment is more for sour iron
Deficiency.
Specific embodiment
The following is a clear and complete description of the technical scheme in the embodiments of the invention.
Raw material is food grade caseins in following embodiment, is bought in Gansu Hua Ling casein limited liability company.
Embodiment 1
A kind of preparation method of iron protein succinylate, includes the following steps：
1.2Kg purified water is added in the reaction flask of 5L, is added with stirring casein 100g.It finishes, stirs 0.5 hour,
2.5mol/L sodium hydroxide solution is added while stirring, adjusting pH stable to 7.5 dissolves casein.Again to casein solution
In total 300g succinic anhydride is added points for 20 times, a hour, which is added, to be finished, and 2.5mol/L hydrogen is added when succinic anhydride is added
Sodium hydroxide solution makes reacting liquid pH value be maintained at 7.5.Succinic anhydride finishes, and continues after stirring 30min, the salt of 3mol/L is added
Acid adjusts the pH value of reaction solution to 2.5, continues after stirring 30min, filtering obtains succinyl albumen wet solid.By succinyl albumen
Wet solid is added in 5L reaction flask, and 500g purified water is added, and 2.5mol/L sodium hydroxide solution is added while stirring and adjusts pH
Value makes succinyl protein dissolution, filters, gained filtrate adds appropriate amount of water, makes succinic acid protein solution total volume 600ml to 4.5.
It takes solution appropriate, measures succinyl protein content, and measure succinyl albumen acylation degree.
Wherein, succinyl protein by weight measuring method is as follows:
This step succinyl protein solution 10ml is taken, 0.1mol/L hydrochloric acid is added dropwise and adjusts pH=2.0~3.0, filters, gained is solid
At 50~55 DEG C, phosphorus pentoxide is dried under reduced pressure 6 hours body.
Calculation formula：W1=(m/10) × V；
W1：Succinyl protein by weight g；
V：Succinyl protein solution total volume ml；
m：The weight g of gained sample after processing.Following embodiment is same.
The liquor ferri trichloridi of 1.528Kg 9.5wt% is added in upper step succinyl protein solution, finishes and continues to stir
After mixing 30 minutes, filtering obtains iron protein succinylate crude product wet solid.
5L reaction flask is added in iron protein succinylate crude product wet solid obtained by upper step, 1.2Kg purified water is added, stirring is allowed to
At suspension.2.5mol/L sodium hydrate aqueous solution adjusting pH stable is added with stirring to make it completely dissolved 8.5.It is above-mentioned anti-
Answer liquid after 5 μm of filter membrane initial filters, then through 1 μm of filter membrane refined filtration, gained filtrate is added with stirring the salt acid for adjusting pH value of 3mol/L extremely
After 2.5, continue stirring 30 minutes, obtained solid is washed to the pH=7 of eluate, obtains iron protein succinylate wet product, egg by filtering
Bactard acid iron wet product sets first 30 DEG C of dry 0.5h in boiling drier after the granulation of 50 mesh oscillating granulators, then is warming up to 70
DEG C dry 2h, obtains iron protein succinylate 102.7g.
Embodiment 2
A kind of preparation method of iron protein succinylate, includes the following steps：
In the reaction flask of 5L, 1.2Kg purified water is added, is added with stirring casein 100g.It finishes, stirs 0.5 hour,
2.5mol/L sodium hydroxide solution is added while stirring, adjusting pH stable to 7.5 dissolves casein.Into casein solution
Divide 20 times and total 300g succinic anhydride is added, a hour, which is added, to be finished, and 2.5mol/L hydrogen-oxygen is added when succinic anhydride is added
Change sodium solution, reacting liquid pH value is made to be maintained at 7.5.Succinic anhydride finishes, and continues to stir 30min, the hydrochloric acid tune of 3mol/L is added
PH value is saved to 2.5.Continue after stirring 30min, filtering obtains succinyl albumen wet solid.Succinyl albumen wet solid is added to
In 5L reaction flask, 500g purified water is added, 2.5mol/L sodium hydroxide solution is added while stirring and adjusts pH value to 4.2, makes amber
Amber acyl protein dissolution, filtering, gained filtrate add appropriate amount of water, make succinic acid protein solution total volume 600ml.Take solution appropriate,
Succinyl protein content is measured, and measures succinyl albumen acylation degree.
The liquor ferri trichloridi of 1.528Kg 9.5wt% is added in upper step succinyl protein solution, finishes and continues to stir
After mixing 30 minutes, filtering obtains iron protein succinylate crude product wet solid.
5L reaction flask is added in iron protein succinylate crude product wet solid obtained by upper step, 1.2Kg purified water is added, stirring is allowed to
At suspension.It is added with stirring 2.5mol/L sodium hydrate aqueous solution and adjusts pH stable 8.5.Above-mentioned reaction solution is filtered through 5 μm
After film initial filter, then through 1 μm of filter membrane refined filtration, gained filtrate is added with stirring the salt acid for adjusting pH value of 3mol/L to 2.5, continues to stir
30 minutes, obtained solid was washed to the pH=7 of eluate, obtains iron protein succinylate wet product, iron protein succinylate wet product by filtering
After the granulation of 50 mesh oscillating granulators, first 30 DEG C of dry 0.5h, then the 70 DEG C of dry 2h that heat up in boiling drier are set, albumen amber is obtained
Amber acid iron 101.9g.
Embodiment 3
A kind of preparation method of iron protein succinylate, includes the following steps：
In the reaction flask of 5L, 1.2Kg purified water is added, is added with stirring casein 100g.It finishes, stirs 0.5 hour,
2.5mol/L sodium hydroxide solution is added while stirring, adjusting pH stable to 7.5 dissolves casein.Into casein solution
Divide 20 times and total 300g succinic anhydride is added, a hour, which is added, to be finished, and 2.5mol/L hydrogen-oxygen is added when succinic anhydride is added
Change sodium solution, reacting liquid pH value is made to be maintained at 7.5.Succinic anhydride finishes, and continues to stir 30min, the hydrochloric acid tune of 3mol/L is added
PH value is saved to 2.5.Continue after stirring 30min, filtering obtains succinyl albumen wet solid.Succinyl albumen wet solid is added to
In 5L reaction flask, 500g purified water is added, 2.5mol/L sodium hydroxide solution is added while stirring and adjusts pH value to 4.1, makes amber
Amber acyl protein dissolution, filtering, gained filtrate add appropriate amount of water, make succinic acid protein solution total volume 600ml.Take solution appropriate,
Succinyl protein content is measured, and measures succinyl albumen acylation degree.
The liquor ferri trichloridi of 1.528Kg 9.5wt% is added in upper step succinyl protein solution, finishes and continues to stir
After mixing 30 minutes, filtering obtains iron protein succinylate crude product wet solid.
5L reaction flask is added in iron protein succinylate crude product wet solid obtained by upper step, 1.2Kg purified water is added, stirring is allowed to
At suspension.It is added with stirring 2.5mol/L sodium hydrate aqueous solution and adjusts pH stable 8.5.Above-mentioned reaction solution is filtered through 5 μm
After film initial filter, then through 1 μm of filter membrane refined filtration, gained filtrate is added with stirring the salt acid for adjusting pH value of 3mol/L to 2.5, continues to stir
30 minutes, obtained solid was washed to the pH=7 of eluate, obtains iron protein succinylate wet product, iron protein succinylate wet product by filtering
After the granulation of 50 mesh oscillating granulators, first 30 DEG C of dry 0.5h, then the 70 DEG C of dry 2h that heat up in boiling drier are set, albumen amber is obtained
Amber acid iron 80.3g.
Embodiment 4
A kind of preparation method of iron protein succinylate, includes the following steps：
In the reaction flask of 5L, 1.2Kg purified water is added, is added with stirring casein 100g.It finishes, stirs 0.5 hour,
2.5mol/L sodium hydroxide solution is added while stirring, adjusting pH stable to 7.5 dissolves casein.Into casein solution
Divide 20 times and total 300g succinic anhydride is added, a hour, which is added, to be finished, and 2.5mol/L hydrogen-oxygen is added when succinic anhydride is added
Change sodium solution, reacting liquid pH value is made to be maintained at 7.5.Succinic anhydride finishes, and continues to stir 30min, the hydrochloric acid tune of 3mol/L is added
PH value is saved to 2.5.Continue after stirring 30min, filtering obtains succinyl albumen wet solid.Succinyl albumen wet solid is added to
In 5L reaction flask, 500g purified water is added, 2.5mol/L sodium hydroxide solution is added while stirring and adjusts pH value to 4.8, makes amber
Amber acyl protein dissolution, filtering, gained filtrate add appropriate amount of water, make succinic acid protein solution total volume 600ml.Take solution appropriate,
Succinyl protein content is measured, and measures succinyl albumen acylation degree.
The liquor ferri trichloridi of 1.528Kg 9.5wt% is added in upper step succinyl protein solution, finishes and continues to stir
After mixing 30 minutes, filtering obtains iron protein succinylate crude product wet solid.
5L reaction flask is added in iron protein succinylate crude product wet solid obtained by upper step, 1.2Kg purified water is added, stirring is allowed to
At suspension.It is added with stirring 2.5mol/L sodium hydrate aqueous solution and adjusts pH stable 8.5.Above-mentioned reaction solution is filtered through 5 μm
After film initial filter, then through 1 μm of filter membrane refined filtration, gained filtrate is added with stirring the salt acid for adjusting pH value of 3mol/L to 2.5, continues to stir
30 minutes, obtained solid was washed to the pH=7 of eluate, obtains iron protein succinylate wet product, iron protein succinylate wet product by filtering
After the granulation of 50 mesh oscillating granulators, first 30 DEG C of dry 0.5h, then the 70 DEG C of dry 2h that heat up in boiling drier are set, albumen amber is obtained
Amber acid iron 102.8g.
Embodiment 5
A kind of preparation method of iron protein succinylate, includes the following steps：
In the reaction flask of 5L, 1.2Kg purified water is added, is added with stirring casein 100g.It finishes, stirs 0.5 hour,
2.5mol/L sodium hydroxide solution is added while stirring, adjusting pH stable to 7.5 dissolves casein.Into casein solution
Divide 20 times and total 300g succinic anhydride is added, a hour, which is added, to be finished, and 2.5mol/L hydrogen-oxygen is added when succinic anhydride is added
Change sodium solution, reacting liquid pH value is made to be maintained at 7.5.Succinic anhydride finishes, and continues to stir 30min, the hydrochloric acid tune of 3mol/L is added
PH value is saved to 2.5.Continue after stirring 30min, filtering obtains succinyl albumen wet solid.Succinyl albumen wet solid is added to
In 5L reaction flask, 560g purified water is added, 2.5mol/L sodium hydroxide solution is added while stirring and adjusts pH value to 4.5, makes amber
Amber acyl protein dissolution, filtering, gained filtrate add appropriate amount of water, make succinic acid protein solution total volume 700ml.Take solution appropriate,
Succinyl protein content is measured, and measures succinyl albumen acylation degree.
The liquor ferri trichloridi of 1.528Kg 9.5wt% is added in upper step succinyl protein solution, finishes and continues to stir
After mixing 30 minutes, filtering obtains iron protein succinylate crude product wet solid.
5L reaction flask is added in iron protein succinylate crude product wet solid obtained by upper step, 1.2Kg purified water is added, stirring is allowed to
At suspension.It is added with stirring 2.5mol/L sodium hydrate aqueous solution and adjusts pH stable 8.5.Above-mentioned reaction solution is filtered through 5 μm
After film initial filter, then through 1 μm of filter membrane refined filtration, gained filtrate is added with stirring the salt acid for adjusting pH value of 3mol/L to 2.5, continues to stir
30 minutes, obtained solid was washed to the pH=7 of eluate, obtains iron protein succinylate wet product, iron protein succinylate wet product by filtering
After the granulation of 50 mesh oscillating granulators, first 30 DEG C of dry 0.5h, then the 70 DEG C of dry 2h that heat up in boiling drier are set, albumen amber is obtained
Amber acid iron 105.2g.
Embodiment 6
A kind of preparation method of iron protein succinylate, includes the following steps：
In the reaction flask of 5L, 1.2Kg purified water is added, is added with stirring casein 100g.It finishes, stirs 0.5 hour,
2.5mol/L sodium hydroxide solution is added while stirring, adjusting pH stable to 7.5 dissolves casein.Into casein solution
Divide 20 times and total 300g succinic anhydride is added, a hour, which is added, to be finished, and 2.5mol/L hydrogen-oxygen is added when succinic anhydride is added
Change sodium solution, reacting liquid pH value is made to be maintained at 7.5.Succinic anhydride finishes, and continues to stir 30min, the hydrochloric acid tune of 3mol/L is added
PH value is saved to 2.5.Continue after stirring 30min, filtering obtains succinyl albumen wet solid.Succinyl albumen wet solid is added to
In 5L reaction flask, 500g purified water is added, 2.5mol/L sodium hydroxide solution is added while stirring and adjusts pH value to 4.9, makes amber
Amber acyl protein dissolution, filtering, gained filtrate add appropriate amount of water, make succinic acid protein solution total volume 600ml.Take solution appropriate,
Succinyl protein content is measured, and measures succinyl albumen acylation degree.
The liquor ferri trichloridi of 1.528Kg 9.5wt% is added in upper step succinyl protein solution, finishes and continues to stir
After mixing 30 minutes, filtering obtains iron protein succinylate crude product wet solid.
5L reaction flask is added in iron protein succinylate crude product wet solid obtained by upper step, 1.2Kg purified water is added, stirring is allowed to
At suspension.It is added with stirring 2.5mol/L sodium hydrate aqueous solution and adjusts pH stable 8.5.Above-mentioned reaction solution is filtered through 5 μm
After film initial filter, then through 1 μm of filter membrane refined filtration, gained filtrate is added with stirring the salt acid for adjusting pH value of 3mol/L to 2.5, continues to stir
30 minutes, obtained solid was washed to the pH=7 of eluate, obtains iron protein succinylate wet product, iron protein succinylate wet product by filtering
After the granulation of 50 mesh oscillating granulators, first 30 DEG C of dry 0.5h, then the 70 DEG C of dry 2h that heat up in boiling drier are set, albumen amber is obtained
Amber acid iron 104.4g.
Embodiment 7
A kind of preparation method of iron protein succinylate, includes the following steps：
In the reaction flask of 5L, 1.2Kg purified water is added, is added with stirring casein 100g.It finishes, stirs 0.5 hour,
2.5mol/L sodium hydroxide solution is added while stirring, adjusting pH stable to 7.5 dissolves casein.Into casein solution
Divide 20 times and total 300g succinic anhydride is added, a hour, which is added, to be finished, and 2.5mol/L hydrogen-oxygen is added when succinic anhydride is added
Change sodium solution, reacting liquid pH value is made to be maintained at 7.5.Succinic anhydride finishes, and continues to stir 30min, the hydrochloric acid tune of 3mol/L is added
PH value is saved to 2.5.Continue after stirring 30min, filtering obtains succinyl albumen wet solid.Succinyl albumen wet solid is added to
In 5L reaction flask, 900g purified water is added, 2.5mol/L sodium hydroxide solution is added while stirring and adjusts pH value to 4.5, makes amber
Amber acyl protein dissolution, filtering, gained filtrate add appropriate amount of water, make succinic acid protein solution total volume 1000ml.Take solution appropriate,
Succinyl protein content is measured, and measures succinyl albumen acylation degree.
The liquor ferri trichloridi of 1.528Kg 9.5wt% is added in upper step succinyl protein solution, finishes and continues to stir
After mixing 30 minutes, filtering obtains iron protein succinylate crude product wet solid.
5L reaction flask is added in iron protein succinylate crude product wet solid obtained by upper step, 1.2Kg purified water is added, stirring is allowed to
At suspension.It is added with stirring 2.5mol/L sodium hydrate aqueous solution and adjusts pH stable 8.5.Above-mentioned reaction solution is filtered through 5 μm
After film initial filter, then through 1 μm of filter membrane refined filtration, gained filtrate is added with stirring the salt acid for adjusting pH value of 3mol/L to 2.5, continues to stir
30 minutes, obtained solid was washed to the pH=7 of eluate, obtains iron protein succinylate wet product, iron protein succinylate wet product by filtering
After the granulation of 50 mesh oscillating granulators, first 30 DEG C of dry 0.5h, then the 70 DEG C of dry 2h that heat up in boiling drier are set, albumen amber is obtained
Amber acid iron 104.3g.
Embodiment 8
A kind of preparation method of iron protein succinylate, includes the following steps：
In the reaction flask of 5L, 1.2Kg purified water is added, is added with stirring casein 100g.It finishes, stirs 0.5 hour,
2.5mol/L sodium hydroxide solution is added while stirring, adjusting pH stable to 7.5 dissolves casein.Into casein solution
Divide 20 times and total 300g succinic anhydride is added, a hour, which is added, to be finished, and 2.5mol/L hydrogen-oxygen is added when succinic anhydride is added
Change sodium solution, reacting liquid pH value is made to be maintained at 7.5.Succinic anhydride finishes, and continues to stir 30min, the hydrochloric acid tune of 3mol/L is added
PH value is saved to 2.5.Continue after stirring 30min, filtering obtains succinyl albumen wet solid.Succinyl albumen wet solid is added to
In 5L reaction flask, 1.2Kg purified water is added, 2.5mol/L sodium hydroxide solution is added while stirring and adjusts pH value to 7.5, makes amber
Amber acyl protein dissolution, filtering, gained filtrate add appropriate amount of water, so that succinic acid protein solution total volume is 1500ml.Take solution suitable
Amount measures succinyl protein content, and measures succinyl albumen acylation degree.
The liquor ferri trichloridi of 1.528Kg 9.5wt% is added in upper step succinyl protein solution, finishes and continues to stir
After mixing 30 minutes, filtering obtains iron protein succinylate crude product wet solid.
5L reaction flask is added in iron protein succinylate crude product wet solid obtained by upper step, 1.2Kg purified water is added, stirring is allowed to
At suspension.It is added with stirring 2.5mol/L sodium hydrate aqueous solution and adjusts pH stable 8.5.Above-mentioned reaction solution is filtered through 5 μm
After film initial filter, then through 1 μm of filter membrane refined filtration, gained filtrate is added with stirring the salt acid for adjusting pH value of 3mol/L to 2.5, continues to stir
30 minutes, obtained solid was washed to the pH=7 of eluate, obtains iron protein succinylate wet product, iron protein succinylate wet product by filtering
After the granulation of 50 mesh oscillating granulators, first 30 DEG C of dry 0.5h, then the 70 DEG C of dry 2h that heat up in boiling drier are set, albumen amber is obtained
Amber acid iron 106.4g.
Embodiment 9
A kind of preparation method of iron protein succinylate, includes the following steps：
In the reaction flask of 5L, 1.2Kg purified water is added, is added with stirring casein 100g.It finishes, stirs 0.5 hour,
2.5mol/L sodium hydroxide solution is added while stirring, adjusting pH stable to 7.5 dissolves casein.Into casein solution
Divide 20 times and total 300g succinic anhydride is added, a hour, which is added, to be finished, and 2.5mol/L hydrogen-oxygen is added when succinic anhydride is added
Change sodium solution, reacting liquid pH value is made to be maintained at 7.5.Succinic anhydride finishes, and continues to stir 30min, the hydrochloric acid tune of 3mol/L is added
PH value is saved to 2.5.Continue after stirring 30min, filtering obtains succinyl albumen wet solid.Succinyl albumen wet solid is added to
In 5L reaction flask, 500g purified water is added, 2.5mol/L sodium hydroxide solution is added while stirring and adjusts pH value to 7.5, makes amber
Amber acyl protein dissolution, filtering, gained filtrate add appropriate amount of water, make succinic acid protein solution total volume 600ml.Take solution appropriate,
Succinyl protein content is measured, and measures succinyl albumen acylation degree.
The liquor ferri trichloridi of 1.528Kg 9.5wt% is added in upper step succinyl protein solution, finishes and continues to stir
After mixing 30 minutes, filtering obtains iron protein succinylate crude product wet solid.
5L reaction flask is added in iron protein succinylate crude product wet solid obtained by upper step, 1.2Kg purified water is added, stirring is allowed to
At suspension.It is added with stirring 2.5mol/L sodium hydrate aqueous solution and adjusts pH stable 8.5.Above-mentioned reaction solution is filtered through 5 μm
After film initial filter, then through 1 μm of filter membrane refined filtration, gained filtrate is added with stirring the salt acid for adjusting pH value of 3mol/L to 2.5, continues to stir
30 minutes, obtained solid was washed to the pH=7 of eluate, obtains iron protein succinylate wet product, iron protein succinylate wet product by filtering
After the granulation of 50 mesh oscillating granulators, first 30 DEG C of dry 0.5h, then the 70 DEG C of dry 2h that heat up in boiling drier are set, albumen amber is obtained
Amber acid iron 103.7g.
Embodiment 10
A kind of preparation method of iron protein succinylate, includes the following steps：
In the reaction flask of 5L, 1.2Kg purified water is added, is added with stirring casein 100g.It finishes, stirs 0.5 hour,
2.5mol/L sodium hydroxide solution is added while stirring, adjusting pH stable to 7.5 dissolves casein.Into casein solution
Divide 20 times and total 300g succinic anhydride is added, a hour, which is added, to be finished, and 2.5mol/L hydrogen-oxygen is added when succinic anhydride is added
Change sodium solution, reacting liquid pH value is made to be maintained at 7.5.Succinic anhydride finishes, and continues to stir 30min, the hydrochloric acid tune of 3mol/L is added
PH value is saved to 2.5.Continue after stirring 30min, filtering obtains succinyl albumen wet solid.Succinyl albumen wet solid is added to
In 5L reaction flask, 1.2Kg purified water is added, 2.5mol/L sodium hydroxide solution is added while stirring and adjusts pH value to 4.5, makes amber
Amber acyl protein dissolution, filtering, gained filtrate add appropriate amount of water, make succinic acid protein solution total volume 1500ml.Take solution appropriate,
Succinyl protein content is measured, and measures succinyl albumen acylation degree.
The liquor ferri trichloridi of 1.528Kg 9.5wt% is added in upper step succinyl protein solution, finishes and continues to stir
After mixing 30 minutes, filtering obtains iron protein succinylate crude product wet solid.
5L reaction flask is added in iron protein succinylate crude product wet solid obtained by upper step, 1.2Kg purified water is added, stirring is allowed to
At suspension.It is added with stirring 2.5mol/L sodium hydrate aqueous solution and adjusts pH stable 8.5.Above-mentioned reaction solution is filtered through 5 μm
After film initial filter, then through 1 μm of filter membrane refined filtration, gained filtrate is added with stirring the salt acid for adjusting pH value of 3mol/L to 2.5, continues to stir
30 minutes, obtained solid was washed to the pH=7 of eluate, obtains iron protein succinylate wet product, iron protein succinylate wet product by filtering
After the granulation of 50 mesh oscillating granulators, first 30 DEG C of dry 0.5h, then the 70 DEG C of dry 2h that heat up in boiling drier are set, albumen amber is obtained
Amber acid iron 104.1g.
The acylation degree measuring method of the various embodiments described above
The percentage of amino and the initial free amine group of casein that casein is acylated in acylation reaction is referred to as being acylated
Degree.
Ninhydrin solution and amino acid heat together generate ammonia, ammonia and ninhydrin and reproducibility ninhydrin reaction, generate purple
Compound.The depth of color and the content of amino acid are directly proportional, by the absorbance at measurement 570nm wavelength, measure amino acid
Content.
Measuring method：Precision measures casein contrast solution and each 5ml of test solution, sets in 25ml volumetric flask, adds indenes three
Ketone color developing agent 1ml, boiling water bath heat 15 minutes, water-bath cooling to room temperature, and then using volume fraction respectively is 60% ethyl alcohol constant volume
To scale.It places 20 minutes, detects the absorbance at 570nm wavelength, and do blank correction.Acylation degree is calculated as follows：
Asp：Succinyl albumen test solution absorbance；
Ap：Casein reference substance solution absorbance；
Cp：Casein reference substance solution concentration；
Csp：Succinyl albumen test solution concentration.
Above-described embodiment intermediate succinic acyl albumen acylation degree measurement result see the table below：
Table 1：The intermediate succinic acyl albumen acylation degree measurement result of Examples 1 to 10
| Embodiment | Embodiment 1 | Embodiment 2 | Embodiment 3 | Embodiment 4 | Embodiment 5 |
| Acylation degree | 99.5% | 99.6% | 99.7% | 99.5% | 99.3% |
| Embodiment | Embodiment 6 | Embodiment 7 | Embodiment 8 | Embodiment 9 | Embodiment 10 |
| Acylation degree | 98.3% | 96.5% | 95.6% | 96.8% | 96.5% |
Above-described embodiment prepares iron protein succinylate aqueous solution characteristic research
Dissolubility：Iron protein succinylate sample is dissolved in appropriate 2.5mol/L sodium hydroxide solution, and adds water constant volume,
It is prepared into the solution of pH=8, the ratio of the liquor capacity ml after constant volume and iron protein succinylate sample quality g are 10:1.Then
By 6 μm of filter membrane vacuum filters, measured after gained insoluble matter is dry.And its dissolving ratio is calculated according to initial amount to be dissolved.
Viscosity：Iron protein succinylate sample is dissolved in appropriate 2.5mol/L sodium hydroxide solution, adds water dilution, preparation
It is respectively 5%, 6% solution at pH=8, iron protein succinylate mass fraction, then measures its solution viscosity.
Filter membrane passability：5ml mass fraction is the iron protein succinylate solution under 5% viscosity measurement, and vacuum filtration passes through
Diameter 3cm, the filter membrane that aperture is 0.45 μm.
Sediment：Iron protein succinylate sample 8g is weighed, is dissolved in appropriate 2.5mol/L sodium hydroxide solution, is prepared into
The solution of pH=8, and water is added to be settled to 150ml.Above-mentioned solution 60ml is taken, is set in centrifuge tube (pipe diameter 40mm, high 115cm),
4000 revs/min are centrifuged 10 minutes, count bottoms volume.
Iron protein succinylate aqueous solution characteristic result of study prepared in the above embodiments see the table below 2~3：
Table 2：Examples 1 to 10 iron protein succinylate aqueous solution characteristic result of study
Above-described embodiment prepares iron protein succinylate index of correlation measurement result and see the table below：
Table 3：Examples 1 to 10 iron protein succinylate measurement result
| Inspection project | Free iron | Iron content | Free succinic acid | Total succinic acid | Chloride |
| Embodiment 1 | It is not detected | 5.1% | 0.02% | 9.8% | <0.1% |
| Embodiment 2 | It is not detected | 4.9% | 0.03% | 9.6% | <0.1% |
| Embodiment 3 | It is not detected | 5.0% | 0.01% | 9.5% | <0.1% |
| Embodiment 4 | It is not detected | 5.3% | 0.02% | 9.6% | <0.1% |
| Embodiment 5 | It is not detected | 5.2% | 0.02% | 9.7% | <0.1% |
| Embodiment 6 | It is not detected | 4.8% | 0.03% | 9.2% | <0.1% |
| Embodiment 7 | It is not detected | 5.1% | 0.01% | 9.0% | <0.1% |
| Embodiment 8 | It is not detected | 5.4% | 0.03% | 8.8% | <0.1% |
| Embodiment 9 | It is not detected | 5.3% | 0.02% | 8.9% | <0.1% |
| Embodiment 10 | It is not detected | 5.3% | 0.03% | 8.6% | <0.1% |
Claims (6)
1. a kind of preparation method of iron protein succinylate, which is characterized in that include the following steps：
1）Casein and 10 ~ 15 times of weight waters are added in reaction vessel, it is molten to be added with stirring 1mol/L ~ 4mol/L sodium hydroxide
Liquid, which adjusts reaction solution pH=7.0 ~ 10.0, dissolves casein, then it is 1 that total weight and casein weight ratio are added in batches:0.2
~1:0.4 succinic anhydride, while 1mol/L ~ 4mol/L sodium hydroxide solution is added and keeps reaction solution pH=7.0 ~ 10.0, amber
After acid anhydrides is added, continue to be stirred to react 30 ~ 45min, then 1mol/L ~ 5mol/L salt acid for adjusting pH=2 are added into reaction solution
~ 3, continue 30 ~ 45min of stirring, filtering, washing obtain succinyl albumen wet solid；
2）Step（1）Casein inventory 5 ~ 7 times of weight waters dispersion of succinyl albumen wet solid, be added with stirring 1mol/L
~ 4mol/L sodium hydroxide solution, which adjusts reaction solution pH=4.1 ~ 4.8, makes succinyl protein dissolution obtain succinyl after centrifugation or filtering
Protein solution；
3）The liquor ferri trichloridi of 1wt% ~ 10wt % is added to step（2）Succinyl protein solution in, succinyl albumen with
The weight ratio of ferro element is 1:0.45~1:0.55, after addition, continue 30 ~ 45min of stirring, filtering, water washing and precipitating obtain egg
Bactard acid iron crude product wet solid；
4）Step（3）Casein inventory 10 ~ 15 times of weight waters dispersion of iron protein succinylate crude product wet solid, stirring it is lower plus
Entering 1mol/L ~ 4mol/L sodium hydroxide solution adjusting reaction solution pH=7.0 ~ 10.0 dissolves iron protein succinylate crude product, will be above-mentioned
1mol/L ~ 5mol/L salt acid for adjusting pH=2 ~ 3, filtering, institute is added through 5 μm of filter membrane initial filters, then after 1 μm of filter membrane refined filtration in reaction solution
Solid washing is obtained, iron protein succinylate wet product is obtained；
5）By step（4）Gained iron protein succinylate wet product is pelletized with 10 ~ 50 mesh of oscillating granulator, then set in boiling drier
It is dry, obtain iron protein succinylate.
2. a kind of preparation method of iron protein succinylate according to claim 1, it is characterised in that：Step 2）Described in
1mol/L ~ 4mol/L sodium hydroxide solution is added and adjusts reaction solution pH=4.1.
3. a kind of preparation method of iron protein succinylate according to claim 2, it is characterised in that：Step 2）Described in
Filter cake 5 times of weight water dispersions of casein inventory.
4. a kind of preparation method of iron protein succinylate according to claim 3, it is characterised in that：Step（5）Middle drying
Actual conditions be：Elder generation dry 0.5 ~ 1h at 20 ~ 30 DEG C, then 2 ~ 3h is dried after being warming up to 50 ~ 70 DEG C.
5. a kind of preparation method of iron protein succinylate according to claim 4, it is characterised in that：Step（1）It is middle by amber
Amber acid anhydrides divides in 15-25 addition casein solution.
6. the preparation method of -5 any a kind of iron protein succinylates according to claim 1, it is characterised in that：Succinic anhydride
Total weight and casein weight ratio are 3:1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710330921.8A CN108864271B (en) | 2017-05-11 | 2017-05-11 | Preparation method of iron protein succinate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710330921.8A CN108864271B (en) | 2017-05-11 | 2017-05-11 | Preparation method of iron protein succinate |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108864271A true CN108864271A (en) | 2018-11-23 |
| CN108864271B CN108864271B (en) | 2022-01-18 |
Family
ID=64319743
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710330921.8A Active CN108864271B (en) | 2017-05-11 | 2017-05-11 | Preparation method of iron protein succinate |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108864271B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110903379A (en) * | 2019-12-20 | 2020-03-24 | 北京海泰天正医药科技有限公司 | Iron protein succinate and method for industrially preparing iron protein succinate |
| CN111004314A (en) * | 2019-03-22 | 2020-04-14 | 江苏中天药业有限公司 | Preparation method of iron protein succinate with high iron loading capacity |
| CN114634562A (en) * | 2020-12-16 | 2022-06-17 | 武汉科福新药有限责任公司 | Preparation method of iron protein succinate |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102838667A (en) * | 2012-09-25 | 2012-12-26 | 江苏济川制药有限公司 | Preparation method of iron protein succinylate |
| CN104402984A (en) * | 2014-11-11 | 2015-03-11 | 武汉科福新药有限责任公司 | Preparation method of iron-protein succinylate with high iron content |
-
2017
- 2017-05-11 CN CN201710330921.8A patent/CN108864271B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102838667A (en) * | 2012-09-25 | 2012-12-26 | 江苏济川制药有限公司 | Preparation method of iron protein succinylate |
| CN104402984A (en) * | 2014-11-11 | 2015-03-11 | 武汉科福新药有限责任公司 | Preparation method of iron-protein succinylate with high iron content |
Non-Patent Citations (1)
| Title |
|---|
| 周陶鸿: "蛋白琥珀酸铁原料及制剂质量和稳定性研究", 《中国优秀硕士学位论文全文数据库》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111004314A (en) * | 2019-03-22 | 2020-04-14 | 江苏中天药业有限公司 | Preparation method of iron protein succinate with high iron loading capacity |
| CN110903379A (en) * | 2019-12-20 | 2020-03-24 | 北京海泰天正医药科技有限公司 | Iron protein succinate and method for industrially preparing iron protein succinate |
| CN114634562A (en) * | 2020-12-16 | 2022-06-17 | 武汉科福新药有限责任公司 | Preparation method of iron protein succinate |
| CN114634562B (en) * | 2020-12-16 | 2023-09-29 | 武汉科福新药有限责任公司 | Preparation method of protein iron succinate |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108864271B (en) | 2022-01-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108864271A (en) | A kind of preparation method of iron protein succinylate | |
| JP5848240B2 (en) | Crystalline sulfated cellulose II and its production from sulfuric acid hydrolysis of cellulose | |
| CN107849157A (en) | The method for preparing relax more glucose and its intermediate | |
| CN105829352B (en) | Method for reclaiming esterified cellulose ether from mixture of reaction products | |
| CN110105460B (en) | Selenylation carboxymethyl pachyman and preparation method and application thereof | |
| WO2019119339A1 (en) | Copper oxychloride particulate matter and preparation method therefor | |
| CN109456421A (en) | A kind of preparation of water soluble chitosan material and application method | |
| CN106397456B (en) | A kind of composition and its refining methd of the Cefdinir containing high-purity | |
| US20020111501A1 (en) | Process for manufacture of in vivo stain composition | |
| CN106589103A (en) | Preparation method of iron proteinsuccinylate | |
| CN106929001A (en) | A kind of fluorescence SiO2The preparation method of Nano microsphere | |
| EP1120427A1 (en) | Method for the formation of low-substituted hydroxypropyl cellulose particles | |
| JPH07110209B2 (en) | Gelatin manufacturing method | |
| CN107880139A (en) | A kind of preparation method of HES preparation | |
| CN109439018A (en) | The method in small particle color lake is prepared in a kind of continuous flow micro passage reaction | |
| CN108440569A (en) | The preparation method of Ceftriaxone Sodium sphaerocrystal | |
| CN109646454A (en) | The preparation method of isomaltose acid anhydride iron 1000 | |
| CN107090011A (en) | Succinyl casein iron (III) and preparation method thereof | |
| CN109021066A (en) | A kind of method of broth extraction glutathione | |
| CN105985232A (en) | Ferric citrate with high iron content and preparation method therefor | |
| CN109650602B (en) | Method for removing antimony ions in water body by using magnetic adsorption material | |
| JPH03131533A (en) | Production of ruthenium nitrate solution | |
| CN105566620B (en) | Thermo-sensitive gathers（Lactic acid valine）Material and preparation method thereof | |
| CN110903379B (en) | Iron protein succinate and method for industrially preparing iron protein succinate | |
| Osborne | A HYDROLYTIC DERIVATIVE OF THE GLOBULIN EDESTIN AND ITS RELATION TO WEYL'S ALBUMINATE AND THE HISTON GROUP. |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |