30. In the medium filling test, after the medium is sterilized and before filling, it is sterilized and filtered through a filter membrane to observe the aseptic effect of the filter during the sterilization and installation process. Is it feasible?

Answer: The culture medium filling test is an investigation of the degree of sterility assurance in all steps including aseptic filtration. It is recommended to use the culture medium directly in the aseptic filtration and subsequent filling process after preparation. Pay attention to the actual operation Prevent insoluble particles from clogging the filter.

31. The medium filling test year is re-verified twice a year, how many batches each time?

Answer: For the annual re-verification of a certain product, the usual practice is to conduct two medium filling tests every year, one batch each time.

32. The aseptic culture time of the 2005 edition of the Chinese Pharmacopoeia has changed to 14 days. Does the culture test at two temperatures after the simulated filling of the medium need to be extended?

Answer: The total cultivation time should not be less than 14 days. It can be divided into two temperatures (22.5 degrees and 32.5 degrees) for at least 7 days each, or it can be cultured directly at one temperature (22.5 degrees).

33. The confidence limit of the qualification standard for the medium filling test is 95%, and the probability of infection is 0.1%. Please explain in detail which statistical method is used and how to calculate it by looking up the table.

Answer: For a detailed description of the calculation formula, please refer to page 258 of the "Guidelines for Drug Production Verification (2003)" (Chemical Industry Press) compiled by the Drug Safety Supervision Department of the State Food and Drug Administration and the Drug Certification Management Center.

There are two calculation methods. One is to use the approximate value calculation formula of the Poisson distribution, namely

P(X>0)=1-e-Np>0.95

Among them, P is the confidence limit, N is the number of simulated bottles or batches, p=0.1% (probability of contamination)

Another calculation method is the more accurate binomial calculation formula, namely

P(X>0)=1-(1-X)N

where P is the confidence limit, N is the number of simulated bottles or batches, X = 0.1% (probability of contamination)

On page 259 of the book, there is a table of the relationship between simulated dispensing quantity, pollution quantity and pollution probability in one simulated dispensing when the credibility limit is 95%. The simulated dispensing quantity and pollution quantity can be found from this table. The corresponding value of.

34. In the film 59 on page 72 of the lecture notes, what are the differences in the process verification of powder injections, freeze-dried powder injections, and small-volume injections? , Where does it start and where does it end?

Answer: For sterile powder injections, the form of medium filling has some special characteristics. For example, if you want to prepare a simulated sterile powder, use a syringe to add the liquid medium to the bottle after dispensing; or divide the sterile medium powder into the bottle. After finishing, add sterile water for injection with a syringe. But the purpose is still to investigate the degree of sterility assurance of the entire aseptic packaging process.

35. How to verify the tightness of the container?

Answer: Physical and microbiological testing methods are often used to verify the tightness of containers. Physical detection has many advantages, such as high sensitivity, convenient use, rapid detection and low cost. During the validity period of the product, physical testing methods can be used to determine whether the integrity of the package meets the specified requirements. An important reason for packaging integrity testing is to ensure that sterile products are always sterile. Therefore, in the development stage of product packaging, microbial penetration tests should be considered, or physical test methods that have been verified and more effective than microbial detection should be considered to detect the integrity of product packaging. However, for the stability test of the product within the validity period, it is more difficult to carry out the microbial invasion test, so physical testing methods are recommended. The microbial invasion test is a challenging test for the integrity of the terminally sterilized container/sealing system. In the verification test, take an infusion bottle or a vial (vial), fill it with a culture medium, stopper and cap sterilize on a normal production line. Then, the sealing surface of the container is immersed in the high-concentration motility bacterial liquid, taken out, cultivated, and checked for microbial invasion to confirm the integrity of the container sealing system. At the same time, a positive control test is required to confirm the growth-promoting ability of the medium.

36. When using the microbial immersion method to verify the tightness of the container, why should the aluminum cap be removed in advance? After removing the aluminum cap, is there only a rubber stopper left? Will the chemical liquid leak during the test and affect the verification result? ?

Answer: The purpose of removing the aluminum cap is to create a more stringent condition. The freeze-dried powder injection is taken as an example in the lecture. Usually there is a high vacuum in the container, which will not cause leakage. The tester can judge the removal according to the characteristics of the product. Whether the aluminum cover is suitable.

37. In the sealing verification, such as the verification of aluminum barrels, the results cannot be observed with the verification of the culture medium. Is there any other method?

Answer: For containers of sterile raw materials, it is recommended to try physical methods, such as saline infiltration.

38. What is the general sampling volume for each test of tightness verification, and what is the period of re-verification?

Answer: You can take at least 10 bottles from the start, middle, and end from the capping line to test. The initial verification should examine the airtightness at different times within the validity period, and the verification can generally be performed once a year.

39. The verification guidelines have related requirements for the sealing of large infusion products, but there are no requirements for subpackaged and freeze-dried products. Is there no need for sealing verification?

Answer: The container tightness verification should be carried out for large-volume injections, small-volume injections, and powder injections.

40. Does the container tightness test have to be completed in all injection forms such as ampoules and vials?

Answer: The container tightness test must be carried out in all injection forms such as ampoules and vials. However, the methods used are not the same. Ampoules generally use physical testing methods, and vials use physical and microbiological testing methods.