86449-31-6 - Names and Identifiers
86449-31-6 - Physico-chemical Properties
| Physical and Chemical Properties | Ulinastatin is a kind of glycoprotein with the function of protease inhibitor, which was first isolated from human urine, plasmin has a significant effect. At the same time, Ulinastatin in sepsis, severe pancreatitis and other severe inflammatory reactions have a good effect, is considered to be a useful immunomodulator drugs. |
| Use | For protease inhibitors |
86449-31-6 - Standard
Authoritative Data Verified Data
A glycoprotein extracted from fresh human urine of this strain that inhibits the activity of various proteolytic enzymes. Ulinastatin activity per 1 mg of protein should not be less than 3500 units.
Last Update：2024-01-02 23:10:35
86449-31-6 - Legal requirements
Authoritative Data Verified Data
This product should be extracted from the urine of healthy people, and the production process should comply with the requirements of the current version of "good manufacturing practice. In the production process, this product needs to be heated at 60°C for 10 hours to make Virus inactivation.
Last Update：2022-01-01 11:32:41
86449-31-6 - Trait
Authoritative Data Verified Data
- This product is off-white to slightly brown powder; Odorless.
- This product is soluble in water, insoluble in ether.
Last Update：2022-01-01 11:32:41
86449-31-6 - Differential diagnosis
Authoritative Data Verified Data
According to the identification test under the Ulinastatin solution item, the same results were shown.
Last Update：2022-01-01 11:32:41
86449-31-6 - Exam
Authoritative Data Verified Data
loss on drying
take 0.lg of this product and dry under reduced pressure at 60°C for 3 hours, and the weight loss shall not exceed 6.0% (General rule 0831).
hepatitis B surface antigen
take this product, add 0.9% sodium chloride solution to dissolve and dilute the solution containing 100,000 units per lml, according to the kit instructions, should be negative.
acid breakdown, clarity and color of solution, kininogenase substance, molecular weight, related substances, abnormal toxicity, bacterial endotoxin and thrombin-like active substances
According to the method under the Ulinastatin solution check, should comply with the provisions.
Last Update：2022-01-01 11:32:42
86449-31-6 - Titer determination
Authoritative Data Verified Data
take this product, according to the method under the Ulinastatin solution.
Last Update：2022-01-01 11:32:42
86449-31-6 - Category
Authoritative Data Verified Data
Same as Ulinastatin solution.
Last Update：2022-01-01 11:32:43
86449-31-6 - Storage
Authoritative Data Verified Data
sealed and stored at a temperature below 20°C.
Last Update：2022-01-01 11:32:43
86449-31-6 - Ulinastatin solution
Authoritative Data Verified Data
A glycoprotein extracted from fresh human urine of this strain that inhibits the activity of various proteolytic enzymes. The activity of Ulinastatin in each lml should not be less than 100,000 units, and the activity of Ulinastatin in each lmg protein should not be less than 3500 units.
System Requirements
This product should be extracted from the urine of healthy people, and the production process should comply with the requirements of the current version of "good manufacturing practice. In the production process, this product needs to be heated at 60°C for 10 hours to make Virus inactivation.
trait
This product is colorless to yellow clear liquid; Odorless.
identification
- take this product, dilute with water to make a solution containing 1000 units per 1 ml, take 0.5ml, add 0.5ml of 5% phenol solution, shake well, add 2.5ml of sulfuric acid, after shaking, the solution appeared orange-yellow.
- This product was taken and diluted with a 0.2mol / L triethanolamine buffer solution (pH 7.8) for titer measurement to prepare a solution containing 200 units per 1 ml as a test solution. Take one test tube, add 1.6 of the above buffer solution, 0.2ml of the test solution and 0.2 of the trypsin solution for titer determination, shake well, and place it in 25°C water bath for 5 minutes, 1.0 of substrate solution under the item of titer determination, shake well, place in 25°C water bath for 5 minutes, and the solution should be colorless. Another test tube was taken, and 0.2ml of the above buffer was used instead of the test solution. The solution should be yellow in color.
- This product is diluted with water to make a solution containing 2000 units per 1 ml, which is determined by ultraviolet-visible spectrophotometry (General 0401) and has a maximum absorption at a wavelength of 277nm.
- take this product and dilute it with 0.9% sodium chloride solution to make a solution containing 500 units per 1 ml. With boric acid-sodium hydroxide buffer (pH 8.4)(take boric acid 24.736g, add 0.1 mol / L sodium hydroxide solution dissolved and diluted to 1.2%) to prepare 3403 agarose gel plate, according to the immune double diffusion method (general rule), should be with rabbit anti Ulinastatin serum to form a clear precipitation line.
examination
- the pH value of this product is diluted with water to make a solution containing 10000 units per lml, which is determined according to law (General 0631). The pH value should be 6.0~7.5.
- clarity and color of the solution this product is diluted with 0.9% sodium chloride solution to make a solution containing 20000 units per 1 ml, which is checked according to law (General rules 0901 1 and 0902 1), the solution should be clear and colorless; If the color is developed, it should not be deeper than the yellow No. 1 Standard Colorimetric solution.
- kininogenase substance 1. Preparation of substrate solution S-2266 C was taken to be H-D-Val-Leu-Arg-PNA. 2 HC1 ) 25mg, and water was added to dissolve and dilute to prepare a 0.0015mol/L solution, which was stored at 18°C. 2. Preparation of test solution this product is diluted with water to make a solution containing about 50000 units per 1 ml. 3.2 test tubes were used for the determination, 0.4 of human test solution was added for each precision, and then 0.2 mol/L Tris-HCL buffer was added respectively (24.228g of Tris, and of water was added for dissolution, adjust the pH value to 8.0 with 6mol / L hydrochloric acid solution, add water to 0.5 ml), mix well, place in 37°C ± 0.5°C water bath for 5 minutes, and then add glacial acetic acid solution (1-2) in the first tube. 0.1ml, 0.1ml of substrate solution was added to the 2nd tube, and the mixture was shaken immediately and timed. The reaction was carried out in a 37°C ± 0.5°C water bath for 30 minutes, and 0.1ml of substrate solution was added to the 1st tube, the second tube added glacial acetic acid solution (1-2) 0.1, according to UV-visible spectrophotometry (General 0401), with the first tube as the blank, at the wavelength of 405nm, the absorbance of tube 2 should not exceed 0.03.
- the appropriate amount of molecular weight of this product is diluted with water to make a solution containing 2mg of protein per 1 ml. Add the same volume of sample buffer, mix well, place in a water bath for 5 minutes, and let cool, as a test solution; Take an appropriate amount of molecular weight standard (molecular weight of 10000~100000 standard protein) and add a buffer solution for the test sample to make a solution containing 1/xg per 1u1, the solution was placed in a water bath for 5 minutes and allowed to cool as a molecular weight standard solution. As determined by electrophoresis (General rule 0541 fifth reduction method), the concentration of the separating gel was 12.5%, and the gel was stained with Coomassie Brilliant Blue. The molecular weight should be between 37000 and 43000.
- Related substances take this product and dilute it with mobile phase to make a solution containing about 10000 units per 1ml, which is used as a test solution. Take 1ml for precision measurement and put it in a 50ml measuring flask, dilute with mobile phase to the scale, shake, as a control solution; Take the appropriate amount of test solution, heated at 105°C for 3 hours, let cool, add the same volume of test solution, mix well as a system-suitable solution. Using hydrophilic modified silica gel as filler (TSK-GEL-G3000SWxl, 0514 X 7.8mm, 5um or other suitable chromatographic column) according to molecular exclusion chromatography (General rule 300mm); sodium Phosphate buffer (sodium dihydrogen phosphate 6.90g, disodium hydrogen phosphate 17.91g, and sodium chloride 8.77g) was dissolved in 6.8 of water, adjusted to pH 0.7, and added to of water) as mobile phase; The flow rate was per minute; the detection wavelength was 280nm. 20ul of the applicable solution of the system is taken, and human is injected into the liquid chromatograph to record the chromatogram. The separation degree between Ulinastatin peak and adjacent impurity peak shall meet the requirements, and the number of theoretical plates shall not be less than 800 based on Ulinastatin peak. 20 u1 of the test solution and the control solution were respectively injected into the liquid chromatograph, and the chromatogram was recorded to 2 times of the retention time of the main component peak. If there are impurity peaks in the chromatogram of the test solution, the sum of each impurity peak area shall not be greater than the main peak area of the control solution (2.0%).
- hepatitis B surface antigen this product, according to the kit instructions, should be negative.
- abnormal toxicity: take this product, dilute it with sodium chloride injection to make 45000 units of solution containing Ulinastatin per lml, and check it according to law (General rule 1141).
- the amount of endotoxin in Ulinastatin should be less than 1.25EU per 1143 units of Ulinastatin for inspection according to law (General rule 10000).
- thrombin-like active substance 1. Preparation of plasma fresh rabbit blood was placed in a container pre-filled with 3.8% sodium citrate solution (the ratio of sodium valproate solution to blood volume was 1:9) and mixed well, centrifuge at 3500 rpm for 20 minutes at 2-8°C. The supernatant was taken for storage at one 20°C for later use and thawed in a 25°C water bath prior to use. 2. Determination of this product, diluted with barbiturate buffer (pH 7.4) to prepare 5000 units of test solution per 1 ml. Take 2 tubes [(10-12 )mm x 75mm] and add barbiturate buffer (pH 7.4) to the first tube. 0.1 of test solution was added to the second tube, and 0.1 of rabbit plasma was added to the second tube, respectively. The mixture was kept in a 25°C ± 0.5°C water bath for 5 minutes, and 0.37% of 0.1 calcium chloride solution was added rapidly, mix well and time. The time of tube turbidity (initial coagulation) was observed and recorded. The initial setting time of the test tube should not be less than that of the blank control tube.
titer determination
- enzyme activity the appropriate amount of crystalline trypsin (containing 7500 to 10000 BAEE units per 1 mg) was accurately weighed and a cold calcium chloride hydrochloric acid solution (taking 2.94g of calcium chloride, and adding O. OL mol/L hydrochloric acid solution looml dissolved) dissolved and diluted to make each lm l containing 0.lmg solution, immediately with the new system, and ice bath preservation.
- the appropriate amount of benzoyl-L-arginine-P-nitroaniline hydrochloride was taken from the substrate solution, dissolved with water and diluted to prepare a solution containing 1 mg per 1 ml, which was freshly prepared and stored in a dark place.
- preparation of standard solution Ulinastatin standard was taken, and 0.2mol / L triethanolamine buffer (pH 7.8) was added (29.8g of triethanolamine was taken, and of water was added for dissolution, the pH was adjusted to 7.8 with a 4mol / L hydrochloric acid solution, and water was added to dissolve and quantitatively dilute to give a solution containing 50 units per 1 ml.
- preparation of test solution an appropriate amount of this product was accurately measured and quantitatively diluted with 0.2mol/L triethanolamine buffer (pH 7.8) to prepare a solution having the same concentration as the standard solution.
- 0.2ml of 7.8 mol / L triethanolamine buffer (pH 0.1) (preheated to 25°C ± 1.6°C) was placed in the cuvette, add 0.2ml of each standard solution and trypsin solution, mix well, accurately incubate for 5 minutes, keep the temperature in the cuvette at 25°C ± 0.1°C, and add substrate solution (preheated to 25°C ± 0.1°C)1.0ml, immediately shake and time, with water as the blank, according to UV-visible spectrophotometry (General 0401), at the wavelength of 405nm, every 1 minute to measure absorbance, a total of 5 minutes, the rate of change in absorbance should be constant. The absorbance change rate (AAs) per minute was determined by plotting the reaction time on the abscissa and the absorbance on the ordinate.
- the appropriate amount of protein content of this product, precision weighing, according to the determination of protein content (General 0731 first method) determination, that is.
- specific activity units containing Ulinastatin activity per 1 mg of protein were calculated from the measured titer and protein content.
category
protease inhibitors.
storage
sealed and stored at a temperature below 20°C.
Last Update：2022-01-01 11:32:44
86449-31-6 - Ulinastatin for injection
Authoritative Data Verified Data
This product is a sterile lyophilized product of Ulinastatin (solution) plus an appropriate amount of stabilizer and excipient. The potency of Ulinastatin should be 85.0% to 120.0% of the labeled amount.
trait
This product is a white to yellowish lyophilized cake or powder. After reconstitution, it should be colorless to yellow clear liquid with slight opalescence.
identification
take this product, according to the Ulinastatin solution under the identification of (1), (2) test, showed the same results.
examination
- pH value: take this product, add 2ml of water to dissolve, mix well, and measure according to law (General rule 0631). The pH value should be 6.0~7.5.
- loss on drying: Take O.lg of this product, use phosphorus pentoxide as desiccant, and dry under reduced pressure at 60°C for 3 hours. The loss of weight shall not exceed 6.0% (General rule 0831).
- allergic reaction take this product, add sodium chloride injection to dissolve and dilute the solution containing 3000 units per lml, check according to law (General 1147), should comply with the provisions.
- the clarity, color, abnormal toxicity and bacterial endotoxin of the solution should be checked according to the method under Ulinastatin solution.
- others should comply with the relevant provisions under injection (General 0102).
titer determination
take 5 pieces of this product, add appropriate amount of 0.2mol / L triethanolamine buffer (pH 7.8) to dissolve, and transfer the whole amount to the same 100ml measuring flask, dilute to the scale with the above buffer, shake well. Take an appropriate amount of precision, quantitatively dilute with the above buffer to make a solution containing 50 units per 1 ml, and measure and calculate according to the method under Ulinastatin solution.
category
Same as Ulinastatin solution.
specification
(1) 25,000 units (2) 50,000 units (3) 100,000 units
storage
sealed, stored in a cool and dry place.
Last Update：2022-01-01 11:32:45