Insulina iniettabile neutra
Insulin
CAS: 9004-10-8
Molecular Formula: C256H381N65O77S6
Insulina iniettabile neutra - Names and Identifiers
Insulina iniettabile neutra - Physico-chemical Properties
| Molecular Formula | C256H381N65O77S6 |
| Molar Mass | 5793.54364 |
| Density | 1.09 g/cm3 |
| Melting Point | 233 °C |
| Solubility | acidified water, pH2.0: 2mg/mL |
| Appearance | solution |
| Storage Condition | 2-8°C |
| Use | For the treatment of moderate and severe diabetes, malnutrition, early cirrhosis and other diseases |
Insulina iniettabile neutra - Risk and Safety
| Safety Description | S22 - Do not breathe dust. S24/25 - Avoid contact with skin and eyes. |
| WGK Germany | 3 |
| RTECS | NM8900250 |
| FLUKA BRAND F CODES | 3-10 |
Insulina iniettabile neutra - Nature
Open Data Verified Data
- This line is a white or off-white crystalline powder extracted from porcine or bovine pancreas. There are two forms due to different refining methods, one is irregular fine particles, called "amorphous", the other is flat oblique hexahedron. Insulin is soluble in less than 80% of dilute alcohol, dilute acetone, acid, alkali, insoluble in more than 90% of ethanol and other organic solvents, more stable at pH 2~4, it is easy to be damaged in alkaline medium and is variable by heating. In case of protease, strong acid, strong alkali can be destroyed. Isoelectric point pH = 5.3~5.4 easy with zinc, cadmium, cobalt and other divalent metal combination, medicinal often made of zinc insulin, and protamine can form a complex, complex hormone action time is longer.
- the insulin molecule consists of 51 amino acid residues and is divided into A chain (containing 21 amino acid residues) and B chain (containing 30 amino acid residues), the two chains of the mountain two disulfide bonds are connected, A chain itself also has A: sulfur bond.
- The main function of insulin is to regulate glucose metabolism, reduce gluconeogenesis, promote glycogen synthesis, inhibit glycogen decomposition, and accelerate anaerobic glycolysis and aerobic oxidation of glucose, it promotes the utilization of glucose by tissues and promotes the conversion of glucose into fat, thus reducing blood sugar. In addition, it can promote the synthesis of fat, inhibit the decomposition of fat, reduce the production of ketone bodies, and correct the symptoms of ketosis and acidemia. And can promote the synthesis of protein, inhibit the decomposition of protein. Together with glucose, potassium ions can be promoted from the extracellular fluid into the intracellular fluid. In the body can be hydrolyzed by proteolytic enzymes lose activity, 'General injection of drugs, in the blood and tissues are also hydrolyzed by enzymes, the efficacy is short.
Last Update：2024-01-02 23:10:35
Insulina iniettabile neutra - Preparation Method
Open Data Verified Data
- acid alcohol extraction reduced pressure concentration method. Insulin is soluble and stable in about 70% acidic ethanol, whereas proteolytic enzymes in the pancreas are sparingly soluble in this environment and their activity is inhibited. After extracting insulin with acidic ethanol solution, basic and acidic proteins were removed by adjusting the pH value of the extract. The extract was then distilled off with ethanol at low temperature, separated from fat, and salted out with sodium chloride to obtain crude insulin. The pH value of the crude product is adjusted in an appropriate concentration of acetone aqueous solution to further remove the impurity protein. 2n2 + is added to the citric acid buffer solution, and then the fine product of insulin is obtained by recrystallization.
- fractional extraction direct zinc salt precipitation method. The frozen pancreas was dissected with a pancreas planing machine, 2.3-2.6 times 86%-88% ethanol and 5% oxalic acid were added, stirred at 10-15C for 3H, and centrifuged. Filter residue with 1 times the amount of 68% ~ 70% ethanol and 0.4% oxalic acid extraction 2H, centrifugal separation. The two extracts were combined. The extract was adjusted to pH 8.0-8.4 (liquid temperature 10-15 ° C.) by adding concentrated aqueous ammonia with stirring, and immediately filtered to remove basic protein to obtain a clear filtrate. The filtrate was acidified with sulfuric acid to PH 3. 6-3.8, cooled to 5 ° C., and allowed to stand for 4H to fully precipitate acidic protein. The supernatant was sucked into a reduced-pressure concentration tank, and the lower layer was filtered with a canvas. The filtrate was incorporated into the supernatant, and ethanol was distilled off under reduced pressure at 30 ° C. Or lower, and concentrated to 1/9 to 1/10 of the original volume. The concentrated solution was transferred to the fat-removing pan, and immediately cooled to 5 ℃ with ice saline after rapid heating to 50 ℃, and allowed to stand for 3-4H, the lower clear liquid (the upper lipid layer can also recover insulin) is separated, the pH is adjusted to 2. 3~2.5, and 27g/L solid sodium chloride is added at 20~25 ℃ under stirring, stand for about 2H. The precipitation of the salt is the crude insulin. The salting-out material is calculated as the dry weight, 7 times the amount of distilled water is added to dissolve, then 3 times the amount of cold acetone is added, and the pH is adjusted to 4. 2~4.3 with 4mol/L ammonia water, and then acetone is added, the ratio of water and acetone in the solution was adjusted to 7:3. The solution was allowed to cool to below 5 ° C. The next day, centrifuged at low temperature, or the precipitate was filtered off to obtain a filtrate in which acidic protein was removed. 4mol/L ammonia water was added to the filtrate to bring the pH to 6.2-6.4, and 3. 6% of 20% zinc acetate solution was adjusted to pH 6.0 with 4mol/L ammonia water. The solution was kept at low temperature overnight and filtered the next day to separate the precipitate. The precipitate was washed with cold acetone to obtain a dry product (0.1 ~ 0.125g dry product per kg pancreas). Add 50ml of ice-cold 2% citric acid, 2ml of 65% zinc acetate solution and 16ml of acetone per gram of dry weight, and dilute to lOOmL with ice water to fully dissolve. Then, it was cooled to below 5 ° C., adjusted to pH 8.0 with 4mol/L aqueous ammonia, and rapidly filtered. The basic protein was removed from the filtrate. The filtrate was immediately adjusted to pH 6 with 10% citric acid solution and supplemented with acetone to maintain the acetone content of 16% throughout the solution system. Then slow stirring 3 ~ sh, the crystallization is precipitated, and then transferred to about 5 deg C cold room for 3-4d, the crystallization is complete. The crystals were collected by centrifugation, and the gray-yellow amorphous precipitate was carefully removed from the upper layer, washed with distilled water or ammonium acetate solution, dehydrated with acetone and ether, centrifuged and dried to obtain insulin crystals.
Last Update：2022-01-01 11:12:01
Insulina iniettabile neutra - Standard
Authoritative Data Verified Data
This line is a protein consisting of 51 amino acid residues extracted from porcine pancreas. Based on the dry product, containing insulin (including deaminating insulin) should be 95.5% to 105.0%.
Each unit corresponds to 0.0345mg of insulin.
Last Update：2024-01-02 23:10:35
Insulina iniettabile neutra - Trait
Authoritative Data Verified Data
- This product is white or off-white crystalline powder.
- This product is almost insoluble in water and ethanol; Soluble in inorganic acid or sodium hydroxide solution.
Last Update：2022-01-01 15:37:52
Insulina iniettabile neutra - Use
Open Data Verified Data
hypoglycemic drugs. Treatment of insulin-dependent diabetes mellitus (diabetes mellitus, type I) and non-insulin-dependent diabetes mellitus (diabetes mellitus, type II) that has not been treated with diet modifications and oral hypoglycemic agents, as well as with severe infections, wasting diseases (such as tuberculosis) diabetic retinopathy, neuropathy, nephropathy, pregnancy and major surgery of various types of diabetes; Treatment of ketoacidosis and hyperosmolar nonketotic rash diabetes Coma; Correction of intracellular potassium deficiency.
Last Update：2022-01-01 11:12:02
Insulina iniettabile neutra - Differential diagnosis
Authoritative Data Verified Data
- in the chromatogram recorded under the content determination item, the retention time of the main peak of the test solution should be consistent with the retention time of the main peak of the reference solution.
- take an appropriate amount of this product, add 0.1% trifluoroacetic acid solution to dissolve and dilute to make a solution containing 10 mg per 1 ml, take 20 u1, add 0.2mol/L Tris-HCL buffer (pH 7.3)20ul, 0.1% V8 enzyme solution 20u1 and water 140 u1, mix well, place in 37°C water bath for 2 hours, add 3ul of phosphoric acid as test solution; Take appropriate amount of insulin reference substance, prepare with the same method, as a control solution. Under the chromatographic conditions of content determination, 0.2mol/L sulfate buffer (pH2.3)-acetonitrile (90:10) as mobile phase A, acetonitrile-water (50:50) for mobile Phase B, gradient elution was performed as follows. Take 25 u1 of each of the reference solution and the test solution, and inject the human liquid chromatograph respectively, and record the chromatographic circle. The peptide map of the test solution should be consistent with the peptide map of the control solution.
Last Update：2022-01-01 15:37:53
Insulina iniettabile neutra - Safety
Open Data Verified Data
It was kept under -15 ℃, closed and protected from light.
Last Update：2022-01-01 11:12:02
Insulina iniettabile neutra - Exam
Authoritative Data Verified Data
Related proteins
take an appropriate amount of this product and add O.Olmol/L hydrochloric acid solution was dissolved and diluted to prepare a solution containing about 3.5mg per 1 ml as a test solution (freshly prepared, stored at 10°C or less). According to the chromatographic conditions under the content determination item, 0.2mol/L sulfate buffer (pH 2.3)-acetonitrile (82:18) as mobile phase A, gradient elution was carried out using acetonitrile-water (50:50) as mobile phase B as follows. The mobile phase ratio was adjusted so that the retention time of the insulin peak was about 25 minutes. 20ul of the sample solution was injected into the liquid chromatograph, and the chromatogram was recorded. Calculated by peak area normalization method, A21 deammoniated insulin should not be more than 5.0%, and other related proteins should not be more than 5.0%.
high molecular protein
take an appropriate amount of this product, add O.Olmol/L hydrochloric acid solution to dissolve and dilute to prepare a solution containing about 4mg per 1 ml as a test solution. Tested according to size exclusion chromatography (General 0514). Hydrophilic modified silica gel was used as filler (3 ~ lOurn ); Glacial acetic acid-acetonitrile-0.1% arginine solution (15:20:65) was used as mobile phase r flow rate was 0.5ml per minute; the detection wavelength was 276nm. Take the insulin monomer-dimer control (or take the appropriate amount of insulin and place it at 60°C overnight), and add O.Olmol/L hydrochloric acid solution is dissolved and diluted to make a solution containing about 4mg per lml, and lool is injected into the liquid chromatograph. The resolution of insulin monomer peak and dimer peak should meet the requirements. The sample solution 100 u1 was injected into the liquid chromatograph and the chromatogram was recorded. Excluding other peak areas where the retention time is greater than the insulin peak, the sum of all peak areas where the retention time is less than the insulin peak shall not be greater than 1.0% calculated by peak area normalization method.
loss on drying
take this product about 0.2g, precision weighing, drying to constant weight at 105°C, loss of weight shall not exceed 10.0% (General rule 0831).
zinc
take an appropriate amount of this product and weigh it accurately, add O.Olmol/L hydrochloric acid solution to dissolve and quantitatively dilute to prepare a solution containing about 0.1 mg per 1 ml as a test solution. Another precise amount of zinc single element standard solution (each lml containing zinc lOOOug) appropriate amount, with O.Olmol/L hydrochloric acid solution was quantitatively diluted to produce 0.2ug, 0.4ug, 0.8ug, l of zinc per 1 ml. Oug and 1.2ug of zinc standard solution. The absorbance was measured at a wavelength of 0406 NM according to Atomic Absorption Spectrophotometry (General Rule 1, Method 1). The zinc content shall not exceed 1.0% based on the dry product.
bacterial endotoxin
take this product, add O.Olmol/L hydrochloric acid solution was dissolved and diluted with inspection water to make a solution containing 5mg per 1 ml, which was checked according to law (General rule 1143). The amount of endotoxin per 1 mg of insulin should be less than 10EU.
microbial limit
take 0.3g of this product, according to the microbial limit test of non sterile products: microbial count method (General 1105), the total number of aerobic bacteria in lg donor products shall not exceed 300cfu.
biological activity
take an appropriate amount of this product, according to the insulin bioassay (General 1211) test, the number of experimental animals in each group can be halved, the experiment adopts random design, according to the method of bioassay statistics (General 1431), the titer shall not be less than 15 units per lmg, which shall be calculated by the random design method of parallel line determination of medium quantity reaction.
Last Update：2022-01-01 15:37:54
Insulina iniettabile neutra - Content determination
Authoritative Data Verified Data
measured by high performance liquid chromatography (General 0512).
chromatographic conditions and system suitability test
with eighteen alkyl silane bonded silica gel as filler (5 ~ 10um) with 0.2mol/L sulfate buffer solution (anhydrous sodium sulfate 28.4g, add water to dissolve, add phosphoric acid 1.7, the pH value of ethanolamine was adjusted to 2.3, and water was added to 1000ml)-acetonitrile (74:26) as mobile phase; The column temperature was 40 ° C.; The detection wavelength was 214nm. Take the system applicable solution 20m (take the insulin control, plus O.Olmol/L hydrochloric acid solution is dissolved and diluted to make a solution containing about 40 units per 1 ml, and placed for at least 24 hours), injected into the liquid chromatograph, and the chromatogram is recorded, the degree of separation between the insulin peak and the A21 deaminated insulin peak (relative retention time to the insulin peak is about 1.2) should be not less than 1.8, and the tailing factor should be not more than 1.8.
assay
take the right amount of this product, precision weighing, plus O.Olmol/L hydrochloric acid solution was dissolved and quantitatively diluted to make a solution containing about 40 units per 1 mL (ready to use new system, or stored at 2-4°C, used within 48 hours). Accurately, 20u1 was injected into the liquid chromatograph, and the chromatogram was recorded. An appropriate amount of insulin reference substance was taken and determined by the same method. According to the external standard method, the sum of the peak area of insulin and the peak area of A21 deammoniated insulin was calculated.
Last Update：2022-01-01 15:37:54
Insulina iniettabile neutra - Category
Authoritative Data Verified Data
hypoglycemic drugs.
Last Update：2022-01-01 15:37:55
Insulina iniettabile neutra - Storage
Authoritative Data Verified Data
light-shielding, hermetically sealed, and stored at a temperature below 15°C.
Last Update：2022-01-01 15:37:55
Insulina iniettabile neutra - Legal requirements
Authoritative Data Verified Data
This product should be extracted from the frozen pancreas of Quarantine qualified pigs. The production process shall comply with the requirements of the current version of "Good Manufacturing Practice for Pharmaceutical Products.
Last Update：2022-01-01 15:37:55
Insulina iniettabile neutra - Insulin injection
Authoritative Data Verified Data
- This product is a sterile aqueous solution of insulin. The potency should be between 90.0% and 110.0% of the labeled amount.
- This product can add 1.4~0.25g of glycerol and g of phenol per 100ml.
trait
This product is colorless or almost colorless clear liquid.
identification
take this product, according to the item of insulin identification (1) test, showed the same results.
examination
- the pH value should be 6.6 to 8.0 (General 0631).
- relevant protein take this product, add 9.6mol/L hydrochloric acid solution 3u1 to each lml for acidification, mix well and then use it as a test solution, take the appropriate amount of sample solution (about equivalent to 2 units of insulin, h), record the chromatogram, subtract the phenol peak, and calculate according to the peak area normalization method, the A21 deaminated insulin peak shall not be greater than 5.0%; The sum of peaks of other related proteins shall not be greater than 6.0%.
- high molecular protein take this product, add 9.6mol/L hydrochloric acid solution for 3m to each lml for acidification, mix well and then use it as the test solution; Take the test solution for lool, determine according to the method under the item of insulin, subtract the other peaks whose retention time is greater than the main peak of insulin, and calculate according to the peak area normalization method, the sum of all peak areas whose retention time is less than the main peak of insulin shall not be greater than 2.0%.
- zinc precision quantity take proper amount of this product, use O. The Olmol/L hydrochloric acid solution is quantitatively diluted to make a solution containing about 4 units per 1 ml. As the test solution, the zinc content per 100 units shall not exceed 40ug according to the method under the item of insulin. Fine weighing appropriate amount of phenol (purity> 99.5%) and adding O.Olmol/L hydrochloric acid solution is dissolved and quantitatively diluted to make a solution containing about 0.25mg of phenol per 1 ml as a phenol control solution; Take an appropriate amount of this product with 0.Olmol/L hydrochloric acid solution was quantitatively diluted to prepare a solution containing about 0.25mg of phenol per 1 ml as a test solution; According to the chromatographic conditions under the content measurement item, the detection wavelength was 270Nm. An appropriate amount of insulin control substance was taken, dissolved and diluted with phenol control solution to make a solution containing about 1 mg of insulin per 1 ml, and 20ul was injected into the liquid chromatograph. The resolution of phenol peak and main peak of insulin should meet the requirements. 20ul of the phenol control solution and 20ul of the test solution were respectively injected into the human liquid chromatograph, the chromatogram was recorded, and the peak area was calculated according to the external standard method. The amount of phenol in each lml should be 2.2~2.8mg. Visible foreign matter this product shall be inspected according to law (General rule 0904), and no visible foreign matter such as metal debris, glass chip, color block, fiber with length or maximum particle size exceeding shall be detected. If fine and visible foreign matter or smoke-like particle column is detected, another 20 (bottles) should be re-tested with the same method, no more than 5 test articles (vials) shall be detected from tiny visible foreign matter or smoky particle columns.
- bacterial endotoxin this product, according to the law to check (General 1143), the amount of endotoxin per 1 unit of insulin should be less than 0.80EU.
- others should comply with the relevant provisions under injection (General 0102).
Content determination
precision take the right amount of this product, add 9.6mol/L hydrochloric acid solution per lml 3-acidification, with 0.Olmol/L hydrochloric acid solution is quantitatively diluted to make a solution containing 40 units per 1 mL (ready to use a new system, or stored at 2~4°C, used within 48 hours), as a test solution, the assay was performed according to the method under insulin. According to the external standard method, the sum of the area of insulin peak and A21 deammoniated insulin peak is calculated.
category
Same as insulin.
specification
(l)3ml:300 units (2)10ml:400 units (3)10ml:800 units
storage
sealed and stored in cold place to avoid freezing.
Last Update：2022-01-01 15:37:56
Insulina iniettabile neutra - Protamine zinc insulin injection
Authoritative Data Verified Data
- This product is a sterile suspension of insulin containing protamine and zinc chloride, and its potency should be 90.0% to 110.0% of the labeled amount.
- This product contains protamine 100~1.0 mg and zinc 0.2~0.25mg per 1.4 units; Glycerol 0.25 ~ g and benzoic acid g can be added per.
trait
This product is white or white suspension; Shaking should be able to disperse evenly.
identification
take this product, according to the item of insulin identification (1) test, showed the same results.
examination
- the pH value should be 6.9 to 7.3 (General 0631).
- relevant protein take this product, add 3ul of 9.6mol/L hydrochloric acid solution to every lml, and mix well. After the suspension is clarified, it is used as the test solution. Determine according to the method under insulin, record chromatogram, subtract phenol and fish essence protein peak, calculate according to peak area normalization method, A21 deammoniated insulin peak shall not be greater than 5.0%; the sum of other relevant protein peaks should not be greater than 6.0%.
- Take appropriate amount of high molecular protein, add 3ul of 9.6mol/L hydrochloric acid solution to every lml, mix well, after the suspension is clarified, as the test solution; Take the test solution l00ul, determine according to the method under the item of insulin, record the chromatogram, subtract the other peaks whose retention time is greater than the main insulin peak, and calculate according to the peak area normalization method, the sum of all peak areas whose retention time is less than the insulin peak shall not be greater than 3.0%.
- zinc precision quantity take proper amount of this product, use O.Olmol/L hydrochloric acid solution is diluted to prepare a solution containing 0.4 units per 1 ml as a test solution; Determined according to the method under the item of insulin, the zinc content is 100 to 0.20 mg per 0.25 units.
- the amount of insulin in the supernatant is taken, 1500g is centrifuged for 10 minutes, and the supernatant is taken as the test solution. Olmol/L hydrochloric acid solution was dissolved and quantitatively diluted to prepare a solution containing about 1 unit per 1 ml as a reference solution. The supernatant should not contain more than 2.5% insulin as determined by the method described under the item of insulin determination. Fine weighing appropriate amount of phenol (purity> 99.5%) and adding O.Olmol/L hydrochloric acid solution is dissolved and diluted quantitatively to make a solution containing about 0.25mg of phenol per 1 ml, which is used as a phenol control solution. Olmol/L hydrochloric acid solution was quantitatively diluted to prepare a solution containing about 0.25mg of phenol per 1 ml as a test solution; According to the chromatographic conditions under the content measurement item, the detection wavelength was 270Nm. An appropriate amount of insulin control substance is taken, dissolved and diluted with phenol control solution to make a solution containing about 1 mg of insulin per 1 ml, and 20ul is injected into the liquid chromatograph. The resolution between the phenol peak and the main peak of insulin shall meet the requirements. 20ul of phenol control solution and 20ul of test solution were injected into the liquid chromatograph respectively. The chromatogram was recorded and the peak area was calculated according to the external standard method. The amount of phenol in each lml should be 2.2~2.8mg.
- sterile take this product, add 1% sterile ascorbic acid aqueous solution, shake to clarify the solution, after membrane filtration treatment, rinse with pH 7.0 sterile sodium chloride-peptone buffer (not less than 1101 per membrane), with Staphylococcus aureus as positive control bacteria, according to law (general rule), should comply with the provisions.
- bacterial endotoxin this product, according to the law to check (General 1143), the amount of endotoxin per 1 unit of insulin should be less than 0.80EU.
- others should comply with the relevant provisions under injection (General 0102).
Content determination
take this product, add 9.6mol/L hydrochloric acid solution 3~4u1 per lml to make the solution completely clear, take appropriate amount of precision, use O.Olmol/L hydrochloric acid solution is quantitatively diluted to make a solution containing 40 units of insulin per 1 mL (ready to use a new system, or stored at 2~4°C, used within 48 hours), according to the method under the item of insulin determination, according to the external standard method to insulin and A21 deammoniated insulin peak area calculation, that is obtained.
category
Same as insulin.
specification
(l)10ml:400 units (2)10ml:800 units
storage
sealed and stored in cold place to avoid freezing.
Last Update：2022-01-01 15:37:58
Insulina iniettabile neutra - Protamine zinc insulin injection (30R)
Authoritative Data Verified Data
- this strain is a sterile suspension prepared by mixing a solution made of insulin with a suspension made of protamine and insulin in a certain ratio. Its potency should be 90.0% to 110.0% of the labeled amount, of which the soluble insulin titer should be 30% of the labeled amount.
- This product can be added to the appropriate amount of phenol as a bacteriostatic agent.
trait
This product is white or white suspension, should be able to disperse evenly after shaking. Under the microscope, the crystal is rod-shaped, and the vast majority of crystals should not be less than lum, not more than 60um, no polymer exists.
identification
- 3ul of 9.6mol/L hydrochloric acid solution was added to every 1 ml of this product to make it completely clear, and the same result was shown according to the identification (1) Test under the item of insulin.
- in the chromatogram recorded under the phenol check item, the retention time of the phenol peak in the test solution should be consistent with the retention time of the phenol peak in the control solution.
examination
- the pH value should be 6.9 to 7.8 (General 0631).
- Related substances take this product, add 9.6mol/L hydrochloric acid solution 3u1 per lml and mix it evenly, then take the appropriate amount of test solution (equivalent to 2 units of insulin), according to the chromatographic conditions under the item of insulin, the phenol peak, m-cresol peak and fish sperm protein peak are removed, and calculated by peak area normalization method, the sum of the peak areas of A21 deaminated insulin shall not exceed 5.0%, and the other impurities shall not exceed 6.0%.
- high molecular protein take this product, add 3m of 9.6mol/L hydrochloric acid solution to each lml and mix it evenly, then use it as the test solution; Take the test solution, and check it according to the method under insulin, excluding other peak areas where the retention time is greater than the main insulin peak, the retention time is less than the sum of all peak areas of the insulin peak, calculated by area normalization method, and shall not exceed 3.0%.
- take an appropriate amount of zinc, add 3ul of 9.6mol/L hydrochloric acid solution per lml to make it completely clear. Olmol/L hydrochloric acid solution is quantitatively diluted to prepare a solution containing about 0.4 ~ 0.8ug of zinc per 1 ml as the test solution, and the test solution is checked according to the method under the item of insulin, the zinc content per 100 units shall be 10-40ug.
- soluble insulin precise quantity take this product with O. 1mol/L Tris-hydrochloric acid buffer (pH 8.2,25°C ± 1°C) appropriate amount, equal volume mixing, shaking, 25°C ± 1°C for 1 hour, filtering with 0.2um filter, take the filtrate and add 3ul of 9.6mol/L hydrochloric acid solution per 1 ml to acidify it as a soluble insulin test solution, add 3ul of 9.6mol/L hydrochloric acid solution per 1 ml, after the solution is clarified, A total insulin test solution corresponding to the insulin concentration in the soluble insulin test solution was prepared by quantitative dilution with 0.01mol/L hydrochloric acid solution, and was measured according to the method under the content determination item. The soluble insulin content should be between 25.0% and 35.0% of the total insulin content.
- the appropriate amount of phenol was accurately weighed, dissolved and quantitatively diluted with O.Olmol/L hydrochloric acid solution to prepare a solution of appropriate concentration as a reference solution. Take this product, add 9.6mol/L hydrochloric acid solution 3ul per lml, make it completely clear, take appropriate amount of precision, use O.Olmol/L hydrochloric acid solution was quantitatively diluted to prepare a solution containing appropriate concentration of phenol as the test solution. The detection wavelength was 270nm according to the chromatographic conditions under the insulin content measurement. An appropriate amount of insulin reference substance is taken, and the solution of the above reference substance is added to dissolve and dilute to prepare a solution containing 1 mg of insulin per 1 ml, and 20ul is injected into the liquid chromatograph. The resolution of phenol peak and insulin peak shall meet the requirements.
- 20 u1 of phenol reference solution and 20 u1 of test solution are respectively injected into the liquid chromatograph, the chromatogram is recorded, and the peak area is calculated according to the external standard method, the phenol content shall be between 80% and 110% of the labeled amount.
- sterile take this product, add 1% sterile ascorbic acid solution ml shake to clarify the solution, after the membrane filtration method, according to the law (General Principles 1101), should comply with the provisions.
- bacterial endotoxin this product, according to the inspection (General 1143), the amount of endotoxin per 100 units of insulin should be less than 80EU.
- others should comply with the relevant provisions under injection (General 0102).
Content determination
take this product, add 3ul of 9.6mol/L hydrochloric acid solution per lml to make it completely clear, and take appropriate amount for precision, quantitative dilution with 0.01mol/L hydrochloric acid solution to make a solution containing 40 units per 1 mL (ready to use new system), according to the method under the item of insulin, obtained.
category
Same as insulin.
specification
(l)3ml:300 units (2)10ml:400 units
storage
sealed and stored in cold place to avoid freezing.
Last Update：2022-01-01 15:38:01
Insulina iniettabile neutra - Reference Information
| Introduction | insulin is a white or off-white crystalline powder derived from porcine or bovine pancreas. According to different refining methods, there are two forms: irregular fine powder particles (amorphous) and flat oblique hexahedron. Soluble in dilute alcohol, dilute acetone, acid, alkali, insoluble in more than 90% ethanol and other organic solvents, more stable at pH 2-4, easy to destroy in alkaline medium, heating variability. In case of protease, strong acid, strong alkali can be destroyed. Isoelectric point pH 5.3-5.4, easy to combine with zinc, cadmium, cobalt and other divalent metals. Pharmaceutical often made of zinc insulin, and protamine can form a complex, and the effect of a longer time. |
| Use | for the treatment of moderate to severe diabetes, malnutrition, early cirrhosis and other diseases hypoglycemic agents. For insulin-dependent diabetes mellitus; Non-insulin-dependent diabetes mellitus with severe infection, trauma, major surgery and other severe stress; Diabetic ketoacidosis; Diabetes with pregnancy; Malnutrition, weight loss, liver cirrhosis can be at the same time intravenous glucose and small doses of insulin, to facilitate tissue utilization of glucose. Adverse Reactions: hypoglycemia, due to improper use of insulin. Allergic reactions, local allergies to the injection site erythema, papules, induration; Systemic reactions with urticaria or associated with angioneurotic edema, asthma, Dyspnea, occasional hypotension, Shock or even death. Insulin resistance. Fat atrophy at injection site. Fat hyperplasia at injection site. insulin is commonly used in the treatment of diabetes. Diabetes mellitus is a metabolic endocrine disease characterized by hyperglycemia. It is a metabolic disorder such as sugar, fat and protein caused by insufficient insulin secretion and excessive secretion of hyperglycemia. The hypoglycemic effect of insulin is strong, mainly used for insulin dependent, heavy, medium-sized diabetic patients and complicated with ketoacidosis and hyperosmolar non-ketosis Coma and other complications. Because this product is easy to be destroyed by digestive enzymes, it is invalid by oral administration and must be injected. |
| production method | insulin can be extracted from the pancreas of pigs, cattle, sheep and other edible animals. Genetic engineering has been successfully used to clone the human insulin gene into the deoxyribonucleic acid of Escherichia coli, so that a new technology for producing human islets by using bacteria has been developed and has entered the stage of scale-up trial production. This recombinant product human insulin is not limited by natural resources, low cost, will eventually replace the existing pig, bovine insulin. Process for extraction of insulin from germanium pancreas: 1. Stripping, extraction and filtration 2. Ammonification, pressure filtration, acidification and concentration 3. Degreasing and salting out 4. Refining 5. Crystallization, drying method 1, acid and alcohol extraction, decompression and concentration extraction, alkalization, acidification, the frozen pancreas was crushed, 2.3-2.6 times the amount of 86%-88% ethanol and 5% oxalic acid were added, 10-15 deg C stirring extraction 3H, centrifugal, filter residue and then 1 times the amount of 68%-70% ethanol and 0.4% oxalic acid extraction 2H, combined filtrate after separation. The extract was stirred at 10-15 °c and adjusted to pH 8-8.4 with ammonia water. The protein was removed by pressure filtration, acidified to pH 3.6-3.8 in time, cooled to 5 °c, and left for 4H to precipitate acidic protein. Porcine pancreas [ethanol (70%), oxalic acid] →[10-15 ℃] extract [ammonia] →[pH8-8.4] alkaline solution [sulfuric acid, PH 3.6-3.8 →[5 ℃] acidified solution concentration, fat removal and salting-out the supernatant was placed in a reduced pressure concentrated Pan, the lower layer was filtered with canvas, the precipitate was discarded, the supernatant was incorporated into the filtrate, and ethanol was recovered under reduced pressure, concentrate the remainder to a relative density of 1.04-1.06. The concentrated solution was transferred to a fat-removing Pan, heated to 50 ℃, then decreased to 5 ℃, and left for 4H. The lower clear solution was separated, adjusted to pH2-2.5, and stirred at 20 ℃ to add 270g/L solid sodium chloride, heat preservation and static, to get the crude insulin salting out. Acidizing liquid [below 30 ℃] → concentrated liquid [rapid heating and rapid cooling] → fat-free liquid [NaCl, pH2]→ Salting-out material except acidic protein and zinc precipitation, dissolve in 7 times the amount of distilled water, add 3 times the amount of cold acetone, 4mol/L ammonia water to adjust pH 4.2-4.3, add additional acetone, make the ratio of water and acetone to 7:3, fully stir, 5 degrees below overnight. The next day, the filtrate was separated at low temperature to obtain acidic protein. Ammonia water (4mol/L) was added to the filtrate, pH was adjusted to 6.2-6.4, a 3.6% volume fraction of zinc acetate (20%) solution was added, and then the above ammonia water was adjusted to pH 6, and the temperature was kept overnight. The precipitate was collected by filtration and washed with cold acetone. Salting-out material [water, acetone, ammonia water] →[pH4.2-4.3] filtrate [ammonia water, zinc acetate] →[pH6] precipitate basic protein, crystallize and precipitate by adding cold 2% citric acid 50ml, 65g/L zinc acetate solution 2ml, acetone 16ml per gram, dilute to 100ml with ice water, dissolve and cool to below 5 ℃, use ammonia water (same as before). The filtrate was quickly adjusted to pH 6 with citric acid (10%) solution. Additional acetone was added to make acetone content 16%. After stirring at a slow speed for 3-5H, the crystals were precipitated and left at 5 ° C. For 3-4 days. Centrifugation, take the precipitate, remove the upper gray-yellow precipitate, wash with water, acetone, ether dehydration, centrifugal drying the crystals to obtain crystalline insulin. The preparation was recovered by precipitation at pH 4.2. Precipitate [citric acid, zinc acetate, acetone, ammonia] →[pH8, below 5 ℃, filtered] filtrate [citric acid, acetone] →[pH6] Crystal [dry] → crystallized insulin precipitate recovered acid protein (pH 4.2-4.3) precipitate, stirred with cold distilled water (7 times the amount), hydrochloric acid (2mol/L) to adjust pH 2.5, add 3 times the amount of dry product acetone, with ammonia water (2mol/L) to adjust pH 4.2-4.3, fill with acetone to 30%, refrigerated overnight. Below 5 ℃, filter, adjust pH 6.2 with ammonia water (2mol/L), add 20% ml of zinc acetate (1.8), and adjust pH 6 with ammonia water (2mol/L), cold overnight. Filtration, precipitation according to (4) crystallization treatment. Method 2. Fractional extraction direct zinc salt precipitation extraction the frozen pancreas was dissected with a pancreas planing machine and fed with 82% L ethanol in the proportion of pancreas. Add 6mol/L sulfuric acid to adjust pH 2.8-3 at 10-12 ℃, stir and extract for 0.5h, add 6mol/L hydrochloric acid to adjust pH 2, and continue stirring for 2.5h. The residue was separated, 65% ethanol 150 L was added, adjusted to pH 2 with the above hydrochloric acid, extracted with stirring for 2H, separated, and the extracts were combined twice. The porcine pancreas [ethanol, sulfuric acid, HCl]→[pH2.8-3, 10-12 ℃] extract was basified, acidified, the extract was cooled to 0-5 ℃, adjusted to pH 7.8 with concentrated ammonia, and filtered with diatomite plate and frame, I. E. Acidify pH 2.5 with V hydrochloric acid: V sulfuric acid: V water = 4:1:4, siphon the clear liquid after complete precipitation, suspend and filter, add zinc chloride solution to the clear liquid (add 3kg per 100kg pancreas, pH 2.5), take the clear liquid suspension filtration, clear liquid with concentrated ammonia water to adjust pH 6.8, at 5 deg C overnight. The next day, the clear liquid was removed by siphoning and the precipitate was collected by filtration (both at 0-5 °c). Extract [ammonia, water, mixed acid] →[pH7.8-8, pH2.5] filtrate [zinc chloride, ammonia] →[pH2.5-6.8] Precipitation zinc precipitation, degreasing, salting out, the precipitate was dissolved in 5 times the amount of distilled water, adjusted to pH 2.7 with 6mol/L hydrochloric acid, and placed at 12-15 °c. On the next day, the lower clear skim liquid was released, and the upper fat was washed with 5 times of distilled water. The lower clear liquid was recovered for the next batch of zinc precipitation. Skim liquid with 6mol/L hydrochloric acid to adjust pH 2.5,25 deg C into 270g/L sodium chloride salting out to get crude product, dissolved in 20 times the amount of distilled water, and then the above hydrochloric acid to adjust pH 2.5, add 160g/L sodium chloride salting-out, filter, collect salting-out material, dissolve it in 7 times the amount of distilled water, add 3 times the amount of acetone, with 4mol/L ammonia to adjust pH 4.5, it was cooled to 0-5 °c overnight and the precipitate was removed (recovered separately) to obtain a clear solution. Precipitate [HCl, 12-15 °c] →[pH2.7] skim liquid [HCl, NaCl]→[pH2.5] crude product [HCl, NaCl]→[pH2.5] salting-out product [ammonia, acetone, water] →[pH4.5] clear liquid zinc precipitation, crystallization, drying clear liquid with 4mol/L ammonia to adjust pH 6, plus 20% zinc acetate solution (according to 100 pancreas plus 30 ml), A white precipitate was precipitated, filtered, and the precipitate was collected. Crystallize (calculated as the precipitate from 500 of pancreas) according to the following formula: 2% citric acid 20% ml, 6.5 zinc acetate 160 ml, acetone 1000 ml, diluted to ml with distilled water. It was cooled to 0-5 °c, adjusted to pH 8 with 4mol/L aqueous ammonia, and filtered to remove the precipitate. Adjust pH 6 with 10% citric acid, stir crystallization for 2 days, remove the clear liquid by siphoning, collect the crystals by centrifugation, wash twice with distilled water and acetone, and dry in a phosphorus pentoxide vacuum dryer to obtain the finished insulin product. Clear solution [zinc acetate, ammonia] →[pH 6] precipitate [citric acid, acetone, zinc acetate] →[0-5 ℃, ph 6] Crystal [water, acetone] →[P2O5] finished insulin. |
| toxic substance data | information provided by: pubchem.ncbi.nlm.nih.gov (external link) |
Last Update：2024-04-09 20:52:54
