Answer: The degree of microbial contamination-that is, the quantity can be examined in accordance with the microbial limit test method in the Pharmacopoeia; microorganism species can be identified in turn from the following aspects:
When using residual probability method, microbial contamination level of product before sterilization should be detected, including boiling test of contaminated bacteria (e.g. 100 ℃, 15 minutes) and microbial count.
Microbiological laboratory is essential for injection manufacturers, and its basic functions should include microbial limit test of raw and auxiliary materials, microbial contamination test before sterilization, product aseptic test, bacterial endotoxin test, dynamic inspection of production environment (count of air particles, floating bacteria and settling bacteria). Qualified laboratories can also carry out the following work: identification of microorganisms (it is recommended that the central laboratory of the group carry out API identification), calibration of biological indicators (that is, D value determination, it is suggested that the central laboratory of the enterprise group carry out this work).
25:Is it necessary to verify sterilization process for over-sterilization method? What is the difference between product research and development process and verification in the actual production of residual probability method ?
Answer: of course, verification is required. Article 83 of EU CGMP Appendix: all sterilization processes should be verified. Residual probability method is the design of sterilization process, which itself needs to be verified and confirmed.
26:Is over-killing method certain that there is no need for microbial challenge tests?
Answer: The connotation of over-killing method is that the number of microorganisms in product is reduced by 12 logarithms, and microbial challenge test is to prove that the residual probability of microorganisms is not more than 10-6, so over-killing method can not carry out microbial challenge tests.
27:Is the final sterilized product subject to separate equipment verification for each variety applied for registration? Is it possible to verify equipment only once and other varieties can be in common use? Can equipment verification data not be attached to variety verification data, but only for archival reference?
Answer: The final sterilization product does not need to be verified separately for each variety applied for registration. If the registered variety adopts the same or lower sterilization conditions, the equipment verification under the condition of higher temperature sterilization can only be carried out; the equipment verification data for sterilization conditions of the variety should also be attached to verification data of the variety.
28:Can the sterilization process of non-solution dosage form, semi-solid or powder injection calculate a value similar to F0? Can you calculate SAL? What is the specific stipulation?
Answer: Cannot, but can calculate SAL. Refer to the decision-making tree of EU sterilization process selection.
29:Regarding filling test of culture medium, whether it is verified for production line or for the declared products (each product has to be filled in three batches for verification), may I ask?
(1)Can production line verification replace filling test of declared product (similar varieties)?
(2)What kind of medium do you want, thioglycolate and modified Martin?
(3)Whether three batches of filling tests are required for different packaging specifications in GMP inspection?
Answer: (1) The simulated filling test of culture medium should be carried out according to different products, packaging specifications and packaging forms. For example, if product is powder injection, the simulated filling test of medium will usually fill liquid medium first, and then fill aseptic powder of simulated product (such as PEG aseptic powder); if aseptic injection or freeze-dried powder injection, only need to fill liquid medium; if packaging specifications are 2ml, 5ml, 10ml, then even if the same product needs to carry out its own medium simulation filling test. If packaging is in the form of vial bottles or other forms (such as pre-filled syringe, ampoule), simulated filling tests of culture medium should be carried out respectively due to the use of different production lines.
(1) In the specific work, considering that the medium simulation filling test is actually a systematic verification of the whole production line, including production equipment, environment and personnel operation, etc., if the medium simulation filling test is carried out with the worst condition, that is, the bottle is the largest, the filling speed is the slowest, the personnel is the most, the time is longer than the normal production time, etc., then the medium simulation filling test of each product, each specification can also be carried out without the need; if it is not the worst condition, it should be carried out condition, it should be carried out according to the product.
(2) Broad-spectrum medium is generally used to promote the growth of gram-positive, negative, yeast and mold, such as soybean tryptone medium, and anaerobic medium is used only under special circumstances.
(3) Only at the time of the first verification, each specification requires three consecutive successful medium simulation filling tests, followed by two subsequent times a year and one batch of medium simulation filling tests each time.
30: Is it feasible to observe the aseptic effect of the filter in the process of disinfection and installation in the medium filling test, after sterilizing the medium, before filling, and then filtering through the filter membrane?
A: medium filling test is an investigation of the degree of aseptic assurance of all steps, including aseptic filtration. It is recommended that the medium be prepared and directly used in aseptic filtration and subsequent filling processes. In practice, attention should be paid to prevent insoluble particles from blocking the filter.
31:How many batches of medium filling tests are re-validated twice a year each year?
A: for the annual revalidation of a product, the usual practice is to conduct medium filling tests twice a year, each batch.
32:The sterile culture time of the 2005 edition of the Chinese Pharmacopoeia has changed to 14 days. Is it also necessary to extend the culture test at two temperatures after the medium is simulated and filled?
A: the total culture 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, or one temperature (22.5 degrees) for direct culture.
33. Please explain which statistical method is used and how to calculate the table.
Answer: the detailed explanation of the calculation formula, can refer to the State Food and Drug Administration Drug Safety Supervision Department and Drug Certification Administration Center organized by the "Drug production Verification Guide (2003)"(Chemical Industry Press) a book 258.
There are two calculation methods. One is to use the approximate formula of Poisson distribution, that is, P (X >0)=1- e-Np>0.95, where the P is the confidence limit, the N is the number of simulated bottles or batches ,0.1%(pollution probability)
The other method is a more accurate binomial formula, that is, P (X>0)=1-(1- X), where the P is the confidence limit, the N is the number of bottles or batches, and 0.1%(pollution probability)
On page 259 of the book, there is a table on the relationship between simulated packing quantity, pollution quantity and pollution probability in a simulated packing with 95% confidence limit.
34:In the fifty-ninth film on page 72 of the handout, it is mentioned that the differences in medium filling procedures in the similarities and differences in the process verification of powder injections, freeze-dried powder injections and small volume injections are mainly what, how, where and from where to end?
Answer: for aseptic powder injection, the form of medium filling has some particularity, if want to prepare simulated aseptic powder, add liquid medium to bottle with syringe after packing; or divide aseptic medium powder separately, Add sterile injection water with syringe after completion. However, the purpose is to investigate the aseptic guarantee degree of the whole aseptic packing process.
35:How to verify the sealing of containers?
A: physical and microbiological methods are often used to verify container sealing. 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 packaging integrity meets the prescribed requirements. An important reason for packaging integrity testing is to ensure that aseptic products remain sterile.
Therefore, in the research and development stage of product packaging, we should consider using microbial invasion test or physical test method which has been proved and is more effective than microbial detection to detect the integrity of product packaging. However, for the stability test of the product in the effective period, it is difficult to carry out the microbial invasion test, so it is suggested to adopt the physical detection method. Microbial invasion test is a challenging test for the integrity of the final sterilizing container / sealing system. In the verification test, take the infusion bottle or the cillin bottle (vial), fill into the medium, and sterilize the plug and cover on the normal production line. Then, the container sealing surface was immersed in high concentration of moving bacteria solution, and the microorganism invasion was taken out, cultured and checked to confirm the integrity of the container sealing system. At the same time, positive control test was needed to confirm the growth ability of culture medium.
36:In the use of microbial immersion for container sealing verification, why to remove the aluminum cover in advance, after removing the aluminum cover, is there only rubber plug, so in the course of the test will there be a leakage of liquid and affect the verification results?
A: the removal of aluminum cover is to create a more stringent condition. In the lecture, freeze-dried powder injection is taken as an example. Usually, there is a high vacuum in the container, which will not cause leakage. The tester can judge whether the removal of aluminum cover is applicable according to the characteristics of his product.
37:In the validation of sealability, such as the validation of aluminum drums, the results can not be observed by medium verification. Is there any other method?
A: for containers of aseptic raw materials, it is recommended to try physical methods, such as brine infiltration.
38:What is the general amount of sampling per leak validation and what is the period of revalidation?
Answer: can take at least 10 bottles from the beginning, middle, end from the gland line to carry on the test, the starting verification should investigate the sealing property of different time in the validity period, and then verify that it can be carried out once a year.
39:There are requirements in the validation guidelines for sealability verification of large infusion products, but there are no requirements for sub-pack and freeze-dried products. Is there no need for sealability verification?
A: large volume injection, small volume injection, powder injection should be container sealing verification.
40:Are container tightness tests required in all injection formulations, such as ampoules and saline bottles?
A: Container sealing test must be carried out in all injection dosage forms such as ampoules and saline bottles. However, the methods used are different. The physical test method is generally used in ampoules, and the physical and microbiological methods are used in cillin bottles.