Cephalosporin enzyme sterile powder and preparation method thereof

Abstract

The invention discloses a cephalosporin enzyme sterile powder and a preparation method thereof, the preparation method comprises the following steps: S1, preparing a cephalosporin enzyme solution with certain activity, adding the cephalosporin enzyme solution into mannitol, and uniformly stirring to obtain a mixture A, 10 ml of the cephalosporin enzyme solution is added into every 100 g of mannitol; s2, putting the mixture A obtained in the step S1 into a blast drying oven until the mixture A is dried to obtain a mixture B, and grinding the mixture B to obtain cephalosporin enzyme powder; and S3, subpackaging the cephalosporin enzyme powder obtained in the step S2 into penicillin bottles, and carrying out cobalt 60 irradiation sterilization to obtain the cephalosporin enzyme sterile powder. The stability of the prepared sterile powder of the cephalosporin enzyme is higher than that of a traditional liquid preparation of the cephalosporin enzyme, and the preparation cost of the sterile powder of the cephalosporin enzyme is lower than that of traditional freeze-dried powder of the cephalosporin enzyme.

Description

Technical Field

[0001] The present invention relates to the technical field of the preparation of sterile powder of cephalosporinase. Specifically, it relates to a sterile powder of cephalosporinase and a preparation method thereof. Background Art

[0002] When performing sterility tests on cephalosporin products in pharmaceutical factories, cephalosporinase is usually added to destroy the antibacterial properties (neutralize and inactivate) of cephalosporin drugs, so as to achieve the accuracy of sterility tests. At present, there are two commercial products on the market: freeze-dried powder of cephalosporinase and liquid cephalosporinase. Among them, the freeze-dried powder of cephalosporinase needs to be prepared by a freeze dryer, and at the same time, it is necessary to evacuate to remove moisture. The preparation time is up to 40 hours, and the preparation cost is relatively high; although the liquid preparation of cephalosporinase has a much lower production cost than the freeze-dried powder of cephalosporinase, its process is to carry out bacteria removal by filtration and aseptic sub-packaging in a clean environment, but its stability is poor. Its activity decreases by 50% after being stored in a refrigerator at 2-10°C for one year, the solution becomes turbid, and the protein is easy to polymerize, which brings trouble to use. Summary of the Invention

[0003] In view of the above technical problems in the related art, the present invention provides a sterile powder of cephalosporinase and a preparation method thereof, which can solve the above problems.

[0004] To achieve the above technical objectives, the technical solution of the present invention is realized as follows:

[0005] A preparation method of a sterile powder of cephalosporinase includes the following steps:

[0006] S1. Prepare a cephalosporinase solution with a certain activity, and add the cephalosporinase solution to mannitol and stir evenly to obtain a mixture A, wherein 10 ml of the cephalosporinase solution is added to every 100 g of mannitol;

[0007] S2. Place the mixture A obtained in step S1 in a forced-air oven until the mixture A is dried to obtain a mixture B, and grind the mixture B to obtain a powder of cephalosporinase;

[0008] S3. Sub-pack the cephalosporinase powder obtained in step S2 in ampoules and sterilize it by cobalt-60 irradiation to obtain a sterile powder of cephalosporinase.

[0009] Further, the activity of the cephalosporinase solution in step S1 is 20 million u / ml, and the buffer solution for preparing the cephalosporinase solution is 50 mmol / L tris-HCL buffer solution, and the pH of the tris-HCL buffer solution is 8.0.

[0010] Further, the purity specification of the mannitol in step S1 is analytical pure.

[0011] Further, the drying temperature of the blast drying oven in step S2 is 25°C, and the drying time is 24 hours.

[0012] A sterile powder of cephalosporinase is prepared by the above preparation method.

[0013] Further, the sterile powder of cephalosporinase is a white or off-white powder; after dissolving 1 g of the sterile powder of cephalosporinase in 10 ml of purified water, the measured pH is 6.0 - 8.0.

[0014] The beneficial effects of the present invention: The stability of the sterile powder of cephalosporinase prepared by this application is higher than that of the traditional liquid preparation of cephalosporinase, and the preparation cost is lower than that of the traditional freeze-dried powder of cephalosporinase. Detailed implementation manners

[0015] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art belong to the scope of protection of the present invention.

[0016] Embodiment

[0017] Take 100 grams of mannitol (analytical pure), add 10 ml of cephalosporinase solution (20 million U / ml), stir evenly, put it into a blast drying oven at 25°C {LRH-250 type} for 24 hours, and a solid mixture with the protein fixed on mannitol can be obtained (which can not only maintain the activity of the protein, but also avoid the turbidity and aggregation of the protein solution). Mash it and grind it into a powder. Take a small amount for enzyme activity determination. If the enzyme activity determination meets the requirements, it is sub-packed into vials, and 20 - 30K irradiation with cobalt-60 can obtain the sterile powder of cephalosporinase. The 25°C drying technology is adopted, and the drying time is short, which can save a large amount of electric energy.

[0018] When detecting the prepared sterile powder of cephalosporinase, the following items are included:

[0019] (1) Appearance: Pour out the powder and visually observe that it should be a white powder or an off-white powder.

[0020] (2) Color of the solution: Take a quantity of 1 g, place it in a 25-ml Nessler colorimetric tube, add 10 ml of purified water to dissolve it, and compare it with the standard of tube #2 yellow color, and it shall not be deeper.

[0021] (3) Clarity of the solution: Take a quantity of 1 g, place it in a 25-ml Nessler glass colorimetric tube, add 10 ml of purified water to dissolve it, and compare it with tube #1 turbidity tube, and it shall not be thicker.

[0022] (4) Acidity and alkalinity: Take 1 g of the powder, dissolve it in 10 ml of purified water that has been boiled and cooled, and measure it with a pH meter (pHs-3B type). The result should be between 6.0 and 8.0.

[0023] (5) Determination of enzyme activity: According to the "Method for Preparation and Activity Determination of Penicillinase" in the Chinese Pharmacopoeia, the enzyme activity per vial ≥ 2 million U / vial.

[0024] (6) Sterility: According to the sterility test method in the Chinese Pharmacopoeia, the result should be negative.

[0025] The following is the project test form:

[0026] Serial number Item Standard Detection method 1 Appearance White or almost white powder Visual inspection 2 Color of solution ≤Yellow No. 2 Compare with the standard color 3 Clarity of solution ≤1# Compare with the standard turbidity 4 Acidity and alkalinity 6.0～8.0 Determined by pH meter 5 Enzyme activity per vial ≥2,000,000 U Method in Chinese Pharmacopoeia 6 Sterility Negative Method in Chinese Pharmacopoeia

[0027] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A method for preparing a sterile powder of cephalosporinase, characterized in that, It includes the following steps: S1. Prepare a cephalosporinase solution with a certain activity, add the cephalosporinase solution to mannitol and stir evenly to obtain mixture A, where 10 ml of the cephalosporinase solution is added to every 100 g of mannitol; S2. Place the mixture A obtained in step S1 in a forced-air oven until the mixture A is dried to obtain mixture B, and grind mixture B to obtain a powder of cephalosporinase; S3. Dispense the cephalosporinase powder obtained in step S2 into vials and sterilize it by cobalt-60 irradiation to obtain a sterile powder of cephalosporinase.

2. The preparation method of a cephalosporinase sterile powder according to claim 1, wherein, The activity of the cephalosporinase solution in step S1 is 20 million u / ml, the buffer solution for preparing the cephalosporinase solution is 50 mmol / L tris-HCL buffer solution, and the pH of the tris-HCL buffer solution is 8.

0.

3. The preparation method of a cephalosporinase sterile powder according to claim 1, characterized in that, The purity specification of the mannitol in step S1 is analytical pure.

4. The preparation method of a sterile powder of cephalosporinase according to claim 1, characterized in that, The drying temperature of the forced-air oven in step S2 is 25 °C, and the drying time is 24 hours.

5. A sterile powder of cephalosporinase, characterized in that, It is prepared by the preparation method described in any one of claims 1 to 4.

6. The sterile powder of cephalosporinase according to claim 4, characterized in that, The sterile powder of cephalosporinase is a white or off-white powder; after dissolving 1 g of the sterile powder of cephalosporinase in 10 ml of purified water, the measured pH is 6.0 - 8.0.