CN101204576A - Process for preparing cerebroprotein hydrolysate NaCl injection - Google Patents
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Abstract
The invention discloses a method for preparing sodium chloride injection liquid of brain protein hydrolysate.The preparation includes the processes: weighing the sodium chloride of 90g and adding injection water of 3,000ml dissolve the sodium chloride; adding activated carbon, standing and filtering for removing the carbon; adding the injection water to 8,000ml to prepare sodium chloride solution; weighing stock solution of the brain protein hydrolysate of 1,000ml to ultrafilter by 8,000 dalton ultrafiltration membrane; adding the filtering liquid into the sodium chloride and adding the injection water to 10,000ml, stirring and mixing uniformly and inletting nitrogen into mixed liquid; respectively filtering through filteration membrane of 0.45Mum and 0.22Mum, filling, nitrogen-inletting, encapsulating and sterilizing the mixed liquid. The injection liquid provided by the invention can be directly used in clinic and is unnecessary to be diluted again, thus avoiding secondary pollution, improving the safety of the medicine and convenient for the clinical use; the operation is simplified due to one-time ultrafiltration in the production.
Description
One, technical field
The present invention relates to a kind of preparation method of medicine, particularly relate to a kind of preparation method of brain protein hydrolysate sodium chloride injection.
Two, background technology
At present, the main component of known Cerebrolysin Vial is that the mixture of free amino acid and molecular weight are at the small-molecular peptides below 10000, wherein free amino acid accounts for 85%, the animal brain protolysate belongs to the natural neuropeptide of micromolecule, and they are being subjected to broad research as useful neuromodulator.Cerebrolysin Vial class medicine is to be widely used in the functional disease that treatment is caused by cerebral tissue change at present both at home and abroad.Clinically be used for cerebrovascular and cerebral arteriosclerosis, cerebral trauma sequela, cerebral malacia or apoplexy sequela, cerebral hypoplasia, dementia or alzheimer disease, be the neurasthenia of main performance, slight infant cerebral hypoplasia etc. with the hypomnesis.Such medicine can promote that brain cell DNA is synthetic, repair and regeneration, can improve brain metabolism, cerebral function improvement, promotion brain cell maturation, each seed amino acid that improves intelligence and supply with the brain cell reparation and regenerate required.The animal brain protolysate can directly enter in the cranial nerve cell by blood brain barrier, promotes protein synthesis and influences its respiratory chain, and it also has the defencive function of anti-hypoxia.
Begun to produce Cerebrolysin Vial the external seventies and be used for clinically, domestic nineteen ninety-five is by the health ministry approval and begin to produce.In recent years, being on the increase of such medicine both at home and abroad.
One of preparation method of existing Cerebrolysin Vial preparation is: get fresh Medulla sus domestica and remove surperficial mucosa and bloodstain etc., homogenate, the acetone precipitation that adds 5 times of volumes adds equivalent ether washing again and soaked 1 hour after the precipitate and separate, refilter, vacuum drying makes the brain powder; The brain powder with water dissolution after, use pepsin, pancreatin double-enzyme hydrolysis successively, hydrolyzed solution centrifuging and taking supernatant, ebuillition of heated and rapidly cooling get hydrolyzed solution through plate-and-frame filtration, ultrafiltration and concentrating under reduced pressure then; The hydrolyzed solution branch is pulled on Sephadex G-25 gel chromatography column chromatography, liquid is collected at active peak made smart brilliant liquid through concentrating under reduced pressure, 0.45um microporous filter; According to required amount of liquid medicine, drug standard (numbering WS1-XG-25-2000) by the promulgation on the 28th of National Drug Administration's December in 2000 calculates the aminoacid consumption, add corresponding amino acid ligand and make concentrated wiring liquid, rarely then join, the fill finished product, become sterile preparation through steam sterilization.The shortcoming of this method is: (1) equipment investment is big, and the production cycle is long, and production efficiency is low; With the chromatography method effective ingredient of purifying, each process need 30～48 hours, the production cycle is long; Because the production cycle is long, for ensuring drug quality, requirement is carried out Sephadex G-25 gel filtration chromatography in 2～8 ℃ of freezer workshops, and the necessary import of Sephadex G-25 gel chromatography material, cost an arm and a leg, produce 1,000,000 (10ml) brain protein hydrolysate injections per year and calculate, above-mentioned input just reaches more than 2,000,000 yuans; Use concentrating under reduced pressure and centrifugal step, production efficiency is low, is unfavorable for the control to drug quality; (2) use a large amount of organic chemistry solvents, the boiling point of acetone and ether is extremely low, it is inflammable, explosive, deleterious hazardous chemical, ether also is used as anesthetis, uses a large amount of acetone and ether earlier the brain albumen precipitation to be come out to make the brain powder, and this step is abnormally dangerous, produce in must be between hoolivan, operate loaded down with trivial details, contaminated environment, harm health of operators.
Notification number be CN1228082C patent disclosure a kind of preparation method of Cerebrolysin Vial preparation, this method is through repeatedly microporous filter membrane and ultrafilter membrane ultrafiltration, its shortcoming is: it is low that (1) uses ultrafilter membrane to carry out the ultrafiltration and concentration response rate, though this technology has adopted hyperfiltration technique, but what use is that the molecular weight that dams is that 1000 ultrafilter membrane concentrates, make effective active composition brain polypeptide losing in various degree, greatly reduce yield; (2) be 70,000 and 10,000 ultrafilter membrane repeatedly, just obtain ultrafiltrate, operate more loaded down with trivial details through the microporous filter membrane and the molecular weight that dams.
Three, summary of the invention:
The technical problem to be solved in the present invention is: a kind of work simplification, safe in utilization, the preparation method that can be directly used in clinical brain protein hydrolysate sodium chloride injection are provided.
Technical scheme of the present invention:
A kind of preparation method of brain protein hydrolysate sodium chloride injection, carry out according to the following steps during preparation:
(1) weighing 90 gram sodium chloride add the dissolving of 3000ml water for injection, add active carbon, leave standstill, and decarbonization filtering adds water for injection again to 8000ml, is made into sodium chloride solution;
(2) measure Cerebrolysin Vial stock solution 1000ml, get filtrate, filtrate is added in the described sodium chloride solution, add water for injection again, stir, make mix homogeneously, in mixed liquor, feed nitrogen to 10000ml with 8000 dalton's ultrafilter membrane ultrafiltration;
(3) with described mixed liquor through the filter membrane filtration respectively of 0.45 μ m and 0.22 μ m, fill, logical nitrogen, encapsulate again, sterilizing gets final product.
Aminoacid in the described Cerebrolysin Vial stock solution is: Aspartic Acid 2.40～3.60mg/ml, threonine 0.21～0.39mg/ml, serine 0.21～0.39mg/ml, glutamic acid 3.20～4.80mg/ml, glycine 1.20～1.80mg/ml, alanine 2.40～3.60mg/ml, valine 1.60～2.40mg/ml, methionine 0.35～0.65mg/ml, isoleucine 1.60～2.40mg/ml, leucine 4.80～7.20mg/ml, phenylalanine 1.60～2.40mg/ml, lysine 4.80～7.20mg/ml, histidine 1.04～1.56mg/ml, arginine 0.30～1.10mg/ml, tryptophan 0.35～0.65mg/ml and proline 1.60～2.40mg/ml.
Described activated carbon dosage is that per 10,000 ml injection need active carbon 1.5g, and described sterilization is to sterilize 10 minutes under 121 ℃ condition.
Every 100ml contains total amino acids 281～421mg, Cerebrolysin Vial by total nitrogen 55～67mg, sodium chloride 0.855～0.945g in the described mixed liquor, and PH is controlled at 6.9～7.5, and every 1ml contains endotoxin less than 0.5EU.
Product of the present invention is to the requirement of Cerebrolysin Vial stock solution:
Cerebrolysin Vial stock solution, it is the mixture that contains about 16 kinds of free amino acids that healthy Medulla sus domestica is made through enzyme hydrolysis, and contain a small amount of small-molecular peptides, every 1ml Cerebrolysin Vial solution contains free amino acid 28.08～42.14mg, and Cerebrolysin Vial contains small-molecular peptides by total nitrogen 5.49～6.71mg.
1. character: this product is light yellow clear liquid.
2. differentiate:
(1) gets this product 2ml, add allophanamide aldehyde reagent (get copper sulfate 0.75g and potassium sodium tartrate 0.3g, add water 250ml and make dissolving, under agitation add 10% sodium hydroxide solution 150ml) 4ml, answer displaing amaranth.
(2) get this product, according to high performance liquid chromatography (two appendix VD of Chinese Pharmacopoeia version in 2000) test.
The chromatographic condition gel chromatographic columns is a mobile phase with phosphate buffer (get dipotassium hydrogen phosphate 10.63g and potassium dihydrogen phosphate 5.31g, add isopropyl alcohol 50ml, add water to 1000ml, regulate pH value to 7.0 with phosphoric acid), and flow velocity is 1ml/min, and the detection wavelength is 280nm.
Algoscopy is got contrast solution and is injected chromatograph of liquid for each 10ml of trial target solution, the record chromatogram, for the chromatogram of trial target solution should with the chromatogram basically identical of contrast solution.
3. check:
(1) index of refraction of index of refraction this product (two appendix VI of Chinese Pharmacopoeia version in 2000 F) is 1.341～1.343.
(2) pH value should be 6.9～7.5 (two appendix VI of Chinese Pharmacopoeia version in 2000 H).
(3) protein is got this product 5ml, adds 20% sulfosalicylic acid 2ml, and solution should be clarified.
(4) other materials calculate by area normalization method with reference to above-mentioned discrimination method test, for surpassing 5.0% with the inconsistent peak area of contrast solution chromatogram in the trial target solution chromatogram.
(5) bacterial endotoxin is got this product, checks (two appendix XI of Chinese Pharmacopoeia version in 2000 E) in accordance with the law, and every 1ml contains bacterial endotoxin should be less than 1.5EU.
(6) aseptic this product of getting is checked (two appendix XI of Chinese Pharmacopoeia version in 2000 H) in accordance with the law, should be up to specification.
4. assay
(1) aminoacid is got this product, carries out assay determination with suitable amino-acid analyzer, and other gets the reference substance solution that corresponding aminoacid reference substance is made respective concentration, measures with method.Press external standard method with the various amino acid whose content of calculated by peak area.Amino acid whose content should meet the following requirements:
Aspartic Acid 2.40～3.60mg/ml, threonine 0.21～0.39mg/ml, serine 0.21～0.39mg/ml, glutamic acid 3.20～4.80mg/ml, glycine 1.20～1.80mg/ml, alanine 2.40～3.60mg/ml, valine 1.60～2.40mg/ml, methionine 0.35～0.65mg/ml, isoleucine 1.60～2.40mg/ml, leucine 4.80～7.20mg/ml, phenylalanine 1.60～2.40mg/ml, lysine 4.80～7.20mg/ml, histidine 1.04～1.56mg/ml, arginine 0.30～1.10mg/ml, tryptophan 0.35～0.65mg/ml and proline 1.60～2.40mg/ml, total amount is 28.08～42.12mg/ml.
(2) total nitrogen is got this product, measures (two appendix VII of Chinese Pharmacopoeia version in 2000 D, second method) in accordance with the law, and total nitrogen is 5.49～6.71mg/ml.
In order to control product quality, we adopt high performance liquid chromatography and thin layer chromatography that Cerebrolysin Vial stock solution is measured, the Rf value of calculated activity composition, and compare.
1. experimental apparatus and material
LC-6A high performance liquid chromatograph (shimadzu), SPD-6AV UV-detector (shimadzu), XK96-A flash mixer (the new health doctor in Jiangyan City vortex agitator), microscale sampler, Cerebrolysin Vial stock solution sample (to 3 batches), Cerebrolysin Vial stock solution reference substance-cerebrolysin (Austria, lot number 611327), nor-leucine (the biochemical Shanghai of gill company limited), sodium acetate analytical pure, all the other are chromatographically pure, and derivatization reagent is a phenyl isothiocyanate solution.
2. experimental technique
2.1 the chromatographic condition chromatographic column is that (mobile phase stock solution 100ml adds distilled water and is diluted to 1000ml ACCQTagTM for 3.9mm * 150mm, 4 μ m, B is acetonitrile-water (60: 40), and binary gradient elution, flow velocity are 0.8ml/min, 37 ℃ of column temperatures detect wavelength 254nm, and sample size is 1.0 μ l.
2.2 the preparation precision of aminoacid reference substance solution and need testing solution is measured aminoacid reference substance solution 15.00ml, places the 25ml measuring bottle, dilute with water, and standardize solution shakes up, as this experiment aminoacid reference substance solution.Precision is measured Cerebrolysin Vial stock solution sample 1mL and is placed the 10ml measuring bottle, is diluted to scale with 0.1mol/ml hydrochloric acid, shakes up, and this solution is need testing solution.
2.3 nor-leucine 18mg is got in the preparation of nor-leucine inner mark solution, uses the 0.1mol/ml dissolving with hydrochloric acid, and is diluted to 10ml, mass fraction is 1.8mg/ml.
2.4 the derivative reaction precision is measured the aminoacid reference substance solution and each 200 μ L of need testing solution put in the 2 μ l centrifuge tubes, the accurate nor-leucine inner mark solution 20 μ l that add, add each 120 μ L of 1mol/L triethylamine-acetonitrile solution (144: 856) and 0.1mol/L phenyl isothiocyanate solution, in the even 5min (600r/min) of vortex concussion instrument mid point jolting, room temperature is placed 45min, add normal hexane 300 μ l, behind the even 5min (600r/min) of vortex shaker mid point jolting, leave standstill 15min, draw lower floor's solution, with 0.45 μ m membrane filtration.
2.5 thin layer chromatography is measured former fluid samples of three batches Cerebrolysin Vial and import cerebrolysin, the Rf value of calculated activity composition.
Instrument utensil: glass plate (18cm * 18cm, 5cm * 20cm), drying baker, sample applicator (tack microsyringe, capillary tube), development chamber (flat or double flute chromatography cylinder), mortar, glass rod, baking oven; Adsorbent silica gel G, 0.5% 1,2,3-indantrione monohydrate-acetone developer.Cerebrolysin Vial stock solution specimen lot is respectively (0705181,0705182,0705183).
2.5.1 the preparation of silica gel g thin-layer plate
With washing liquid wash clean and oven dry, glass plate requires smooth surface to the glass plate that thin layer is used in advance.Take by weighing and select white, high-purity and have the thin layer chromatography reagent silica gel G powder 16g that certain particle size distributes, add the 35ml distilled water, be stirred to slowly in beaker with glass rod that the silica gel serosity is uniformly dispersed, stickiness is moderate, be poured over then on clean, the exsiccant sloping glass plate, with glass rod silica gel G is promoted to the other end by an end, with finger tip springing flat board, make silica gel G be paved into the uniform thin layer of thickness.Treat that thin sheet surface water separates drying and is placed in the baking oven, treat that temperature rises to 110 ℃ of postactivated 30min.Take out after being cooled to room temperature, place exsiccator standby (note: avoid that thin plate is shock heating, quenching makes the thin layer fracture or come off) in an exhibition layer process.The lamellae of making requires surfacing, and thickness is even.
2.5.2 point sample
Get one of the thin plate for preparing, the 1.5cm place draws a straight line on the distance base, on straight line every 5cm as a mark (dub with pencil, thin layer can not be punctured), totally 2 points.Capillary tube with the 0.5mm diameter is drawn about 5～50 μ g of sample thief amount, the about 1-1.5 μ of point sample volume l, and graded drips, and control point sample spot diameter is no more than 2mm.The cold and hot wind of available hair-dryer alternately dries up sample in the point sample process, also can allow the sample natural drying.
2.5.3 developing solvent and exhibition layer
First developing solvent: n-butyl alcohol: glacial acetic acid: distilled water was respectively 4: 1: 0.8, second developing solvent: n-butyl alcohol: glacial acetic acid: distilled water was respectively 4: 1: 1.2, thin plate point sample one end of point sample is put into the chromatography cylinder that fills exhibition layer solvent, exhibition layer liquid level must not surpass the some line-transect, chromatography cylinder is airtight, and bottom-up exhibition layer is when exhibition layer solvent takes out thin plate when point of sample upwards launches about 10cm, a mark is made with pencil or little pin in the forward position, 60 ℃ of drying in oven or dry.
2.5.4 0.5% 1,2,3-indantrione monohydrate-acetone developer even spraying on thin layer, is put 85 ℃ of baking oven internal heating to chromatography speckle and is manifested.
2.6 peptide figure analysis
Chromatographic column: TSK GEL2000SWXL (7.8*300mm), mobile phase: phosphate buffer is prepared with getting dipotassium hydrogen phosphate 10.63g, potassium dihydrogen phosphate 5.31g, add isopropyl alcohol 50ml, add water and make and be dissolved into 1000ml, regulate PH to 7.0 with phosphoric acid, flow velocity 1.0ml/min, sample size 10 μ l, 25 ℃ of column temperatures, the long 280nm of check stream.
3. result
3.116 plant the free aminoacid content check result
Aminoacid reference substance solution and each 1 μ L of need testing solution of getting behind the derivative reaction measure, and calculate amino acid content by internal standard method respectively with peak area, see Table 1 and Fig. 1.
The various free aminoacid contents of table 1
| Batch 1 | Batches 2 | |
The import cerebrolysin | |
| 1Asp 2Glu 3Ser 4Gly 5His 6Arg 7Thr 8Ala 9Pro 10Val 11Met 12Ile 13Leu 14Phe 15Trp 16Lys | 3.040±0.239 4.074±0.284 0.323±0.068 1.406±0.168 1.313±0.131 0.599±0.065 0.413±0.061 2.834±0.137 2.146±0.134 1.997±0.130 0.421±0.058 2.053±0.194 6.097±0.365 2.125±0.153 0.351±0.039 5.355±0.506 | 2.862±0.246 4.088±0.237 0.343±0.045 1.428±0.164 1.325±0.118 0.620±0.081 0.403±0.094 2.866±0.135 2.125±0.116 2.046±0.148 0.419±0.043 2.070±0.223 6.048±0.354 2.170±0.197 0.356±0.054 5.314±0.410 | 2.923±0.214 4.261±0.347 0.370±0.040 1.389±0.125 1.365±0.111 0.644±0.060 0.438±0.056 2.845±0.122 2.138±0.118 2.072±0.132 0.463±0.052 2.098±0.318 6.039±0.369 2.164±0.218 0.366±0.041 5.312±0.461 | 3.096±0.245 4.027±0.283 0.318±0.068 1.465±0.173 1.371±0.131 0.652±0.064 0.342±0.056 2.887±0.135 2.073±0.134 2.052±0.132 0.570±0.059 2.108±0.191 6.150±0.368 2.179±0.153 0.501±0.038 6.013±0.504 |
3.2 peptide figure analysis chromatographic column: mobile phase: phosphate buffer (is used Na
2HPO
4H
2O 5.31g and NaH
2PO
4H
2O 10.63g adds isopropyl alcohol 50ml, adds water to 1000ml and mixes, and regulates PH to 7.0 with phosphoric acid), flow velocity 1.0ml/min; Sample size 10 μ l, column temperature 25 detects the long 250nm of stream, sees Fig. 2.
3.3 thin layer chromatography is launched through bi-directional secondary, as can be seen, Cerebrolysin Vial and import cerebrolysin all comprise three kinds of bioactive peptide compositions in the sample, it is approaching that both compare each active component, and its difference not statistically significant (P＞0.05) belongs to same overall, and three batches sample, every batch of repeated experiments 8 times is all consistent with import cerebrolysin result, and stable components is seen Fig. 3, Fig. 4, Fig. 5 and table 2.
Table 2 thin layer chromatography free amino acid and micromolecule polypeptide Rf value
| Composition 1 | Composition 2 | |
|
| Cerebrolysin sample 1 sample 2 |
0.723±0.077 0.703±0.086 0.702±0.080 0.706±0.062 | 0.448±0.046 0.454±0.070 0.448±0.079 0.463±0.090 | 0.197±0.053 0.199±0.041 0.183±0.0 0.191±0.059 |
| F P | 0.15 0.930 | 0.08 0.973 | 0.16 0.920 |
According to two-way thin layer chromatography result, calculate 95% range of normal value quality control standard of micromolecule polypeptide Rf value, for control of product quality provides simple and feasible foundation.
Rf value 95% quality control standard computing formula:
Composition 1:0.561～0.847, composition 2:0.304～0.606, composition 3:0.099～0.283
4. conclusion
High pressure lipuid chromatography (HPLC) and two-way thin layer chromatography measurement result show that all three batches sample is consistent with import cerebrolysin bioactive peptide composition; And two-way thin layer chromatography to have expense low, method is easy, fast, characteristics such as accurate help aborning product quality being carried out dynamic monitoring.This product has been established 95% quality control standard, to guarantee stable and reliable product quality.
Positive beneficial effect of the present invention:
1, the present invention directly adds sodium chloride in Cerebrolysin Vial stock solution, the brain protein hydrolysate sodium chloride injection of making, can be directly used in clinical, be adapted to craniocerebral trauma, cerebrovascular disease (cerebral blood supply insufficiency, cerebral infarction) sequela with diseases such as hypomnesis, need not during use to dilute once more, avoided secondary pollution, improved the safety of medicine, convenient clinical use.
2, product of the present invention contains a large amount of free amino acids and low molecular peptide, aminoacid can be in brain metabolism rapidly, the brain insufficiency person is had the auxiliary improvement effect, and for the treatment of potein deficiency, neurasthenia and general protein digestibility malabsorption, effect is obvious.
3, reasonable, the simplification of methodological science of the present invention.The polypeptide molecular weight that needs in the Cerebrolysin Vial is generally between 6000～10000, because ultrafiltration medium micropore is distributed with certain limit, adopt 8000 dalton's direct ultra-filtration concentrated solutions, effective active composition brain polypeptide is lost few, reduce molecular weight greater than the probability that 10000 peptide material exists, improved the yield of effective active composition.Whole process of production only needs a ultrafiltration, just can remove pyrogen fully, and the ultrafiltration time is few, has simplified operating process.
Four, description of drawings
Fig. 1 aminoacid HPLC chromatogram
Fig. 2 Cerebrolysin Vial peptide figure analysis
First sample Cerebrolysin Vial of Fig. 3 and import cerebrolysin bioactive peptide composition two-dimensional chromatography figure
Fig. 4 second batch sample Cerebrolysin Vial and import cerebrolysin bioactive peptide composition two-dimensional chromatography figure
Five, the specific embodiment:
Embodiment: the preparation method of brain protein hydrolysate sodium chloride injection
Weighing 90 gram sodium chloride add the dissolving of 3000ml water for injection, add 1.5 gram active carbons, leave standstill, and decarbonization filtering adds the injection water to 8000ml, is made into sodium chloride solution; Measure 1000ml Cerebrolysin Vial stock solution, wherein, Aspartic Acid 2.40～3.60mg/ml, threonine 0.21～0.39mg/ml, serine 0.21～0.39mg/ml, glutamic acid 3.20～4.80mg/ml, glycine 1.20～1.80mg/ml, alanine 2.40～3.60mg/ml, valine 1.60～2.40mg/ml, methionine 0.35～0.65mg/ml, isoleucine 1.60～2.40mg/ml, leucine 4.80～7.20mg/ml, phenylalanine 1.60～2.40mg/ml, lysine 4.80～7.20mg/ml, histidine 1.04～1.56mg/ml, arginine 0.30～1.10mg/ml, tryptophan 0.35～0.65mg/ml and proline 1.60～2.40mg/ml.
With the above-mentioned stock solution of 8000 dalton's ultrafilter membrane ultrafiltration, filtrate is added in the described sodium chloride solution; Add water for injection again to 10000ml, in solution, feed nitrogen.
Measure each component content in the solution, make every 100ml injection contain total amino acids 281～421mg, Cerebrolysin Vial by total nitrogen 55～67mg, sodium chloride 0.855～0.945g, PH is controlled at 6.9～7.5, the bacterial detection endotoxin, and every 1ml injection contains endotoxin less than 0.5EU.After qualified, with filter membrane respectively filtration, fill, the logical nitrogen of medicinal liquid, by filling in, roll lid through 0.45 μ m and 0.22 μ m; 121 ℃ of sterilizations 10 minutes.Lamp inspection, labeling, packing.
Claims (4)
1. the preparation method of a brain protein hydrolysate sodium chloride injection is characterized in that, carries out according to the following steps during preparation:
(1) weighing 90 gram sodium chloride add the dissolving of 3000ml water for injection, add active carbon, leave standstill, and decarbonization filtering adds water for injection again to 8000ml, is made into sodium chloride solution;
(2) measure Cerebrolysin Vial stock solution 1000ml, get filtrate, filtrate is added in the described sodium chloride solution, add water for injection again, stir, make mix homogeneously, in mixed liquor, feed nitrogen to 10000ml with 8000 dalton's ultrafilter membrane ultrafiltration;
(3) with described mixed liquor through the filter membrane filtration respectively of 0.45 μ m and 0.22 μ m, fill, logical nitrogen, encapsulate again, sterilizing gets final product.
2. the preparation method of injection according to claim 1, it is characterized in that the aminoacid in the described Cerebrolysin Vial stock solution is: Aspartic Acid 2.40～3.60mg/ml, threonine 0.21～0.39mg/ml, serine 0.21～0.39mg/ml, glutamic acid 3.20～4.80mg/ml, glycine 1.20～1.80mg/ml, alanine 2.40～3.60mg/ml, valine 1.60～2.40mg/ml, methionine 0.35～0.65mg/ml, isoleucine 1.60～2.40mg/ml, leucine 4.80～7.20mg/ml, phenylalanine 1.60～2.40mg/ml, lysine 4.80～7.20mg/ml, histidine 1.04～1.56mg/ml, arginine 0.30～1.10mg/ml, tryptophan 0.35～0.65mg/ml and proline 1.60～2.40mg/ml.
3. the preparation method of injection according to claim 1 is characterized in that, described activated carbon dosage is that per 10,000 ml injection need active carbon 1.5g, and described sterilization is to sterilize 10 minutes under 121 ℃ condition.
4. according to the preparation method of each described injection of claim 1 to 3, it is characterized in that, every 100ml contains total amino acids 281～421mg, Cerebrolysin Vial by total nitrogen 55～67mg, sodium chloride 0.855～0.945g in the described mixed liquor, PH is controlled at 6.9～7.5, and every 1ml contains endotoxin less than 0.5EU.
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| CN101732695B (en) * | 2008-11-20 | 2012-11-07 | 深圳四环医药有限公司 | Compound of brain protein hydrolyzate and maleic acid and method for preparing same |
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| CN101732695B (en) * | 2008-11-20 | 2012-11-07 | 深圳四环医药有限公司 | Compound of brain protein hydrolyzate and maleic acid and method for preparing same |
| CN102718857A (en) * | 2012-07-09 | 2012-10-10 | 河北智同医药控股集团有限公司 | Denatured protein powder and brain protein hydrolyzate prepared from same |
| CN102718857B (en) * | 2012-07-09 | 2013-05-22 | 河北智同医药控股集团有限公司 | Denatured protein powder and brain protein hydrolyzate prepared from same |
| CN104096214A (en) * | 2014-06-10 | 2014-10-15 | 广州一品红制药有限公司 | Cerebroprotein hydrolysate freeze-dried powder injection and preparation method thereof |
| CN104096214B (en) * | 2014-06-10 | 2015-12-30 | 广州一品红制药有限公司 | A kind of Cerebrolysin Vial lyophilized injectable powder and preparation method thereof |
| CN106226438A (en) * | 2016-09-20 | 2016-12-14 | 吉林万通药业集团梅河药业股份有限公司 | A kind of method of inspection of brain protein hydrolysate oral liquid |
| CN106226438B (en) * | 2016-09-20 | 2018-09-21 | 吉林万通药业集团梅河药业股份有限公司 | A kind of method of inspection of brain protein hydrolysate oral liquid |
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