2、指南征求稿中關于培養時間至少 14 天，一般采用先在 20-25℃最少培養 7 天，然后在30-35℃的范圍繼續培養 7 天。 也可采用單一的 20-35℃的培養條件，但應有相關的數據證明培養條件有利于微生物的生長。一是能否先30-35℃后20-25℃培養？二是采用單一的 20-35℃培養怎么操作？
1. In the 2015 edition of Chinese pharmacopoeia, the aseptic examination of aerobic bacteria and fungi was changed to tryptone soybean broth medium (TSB). Should the culture medium of simulated filling test be changed?
A: no change is required. Generally, broad-spectrum tryptone soybean broth medium (TSB), fungus, bacteria can grow. Products if ever had anaerobic growth, can use sulfur glycolic acid salt liquid medium (FTM) simulation of the filling to do simulation test in the same conditions, the anaerobic culture typically layered, upper used to isolate the lower air layer, leading to anaerobic bacteria in the upper is not long. Anaerobic bacteria to grow in the lower, but not because of some bottles, such as 2 ml and 5 ml bottle can't create isolation air layer, due to the simulation of positive bottle is not long, so, is not only make the anaerobic bacteria can also be difficult to survive, so most companies are not doing anaerobic culture medium filling simulation test.
2, guidelines for draft of training time at least 14 days, commonly used in the first 20 to 25 ℃ at least 7 days, and then continue in the range of 30-35 ℃ for 7 days. Also can use a single culture conditions of 20-35 ℃, but shall have the relevant data to prove that culture conditions conducive to the growth of microorganisms. One is whether first 30-35 ℃ after 20 to 25 ℃ training? The second is to use a single 20-35 ℃ training how to operate?
Answer: the first question: is the first low temperature fungi cultivation, and high temperature bacteria, for fear that some bacteria at low temperature to high temperature of 30-35 ℃ train would be killed. In addition, fungi grow more slowly than bacteria, so the fungus can be cultured for 7 days first, followed by another 7 days. And the bacteria grow very fast, generally can be seen in 3 days, so it can be cultured later.
The second question: the cultivation of the culture medium filling temperature, can be done in the medium to long after the test, select a 20-35 ℃ temperature for culture, but to control the temperature of the incubator in the plus or minus 2.5 ℃.
3. What is the "worst condition" for simulated filling test of medium? (translated from the medical micro-school, the article was provided by zhou zhen, shu Yang, sichuan province)
A: 2010 version of the drug GMP guidelines: in the design of media fills, the choice of technological condition selection should be the reasonable conditions of "worst", in the worst condition to the technological process, equipment and the whole system. If good results can be obtained under the worst conditions, the reliability of aseptic guarantee is more guaranteed in the actual production which is better than the worst case. The "worst condition" considers the following three aspects:
First, the storage time after facilities, equipment, packaging materials and auxiliary materials are cleaned and sterilized:
Ii. Filling personnel and parameter setting:
Third, interference project design:
Simulation of the actual production process in various types of regular interference and non-conventional interference. On every six months before the media fills, should a stage summarize production status and trend analysis, considering the activities of the representative and intervention, to type and frequency of the interference on the plan. According to the comprehensive consideration of human, machine, material, method, ring and measurement, the most representative "worst condition" is combined.
4. Major personnel changes. Do you need to carry out the simulated filling test of culture medium before you can be officially employed?
A: article 47 of the aseptic drugs appendix: the first validation of the simulated filling test of the culture medium, three consecutive qualified tests shall be conducted for each shift. After major changes in the air purification system, equipment, production technology and personnel, the simulated filling test of culture medium shall be repeated. The simulated filling test of culture medium shall normally be carried out once every half a year according to the production process, at least one batch at a time. Significant changes, certainly need to medium filling simulation experiment was carried out, only participated in filling simulation test personnel, is allowed to enter to the core filling area for normal production operations.
5. After the completion of the simulated filling of the culture medium, should all the cylinder be inverted or only partially inverted during the culture?
A: as long as the culture medium is in contact with all contact surfaces of the cylinder, whether upright, inverted or flat. Industry there are three kinds of practice: one is the medium filling simulation over, the cultivation of the normal, does not need to be inverted, medium during the bottle come into contact with any surface to be medium, it is ok to flip a few times; The second is to fully contact the culture medium with the cylinder and the rubber stopper, and then half of the vertical culture and half of the vertical culture; Third, the culture medium was fully exposed to the cylinder and the rubber stopper of xilin. The culture medium was incubated at 20-25 degrees for 7 days and inverted at 30-35 degrees for 7 days. Guidelines for draft, is the first one is to cultivate, to reverse the filling simulation products, light wave so that the medium contact all inner surface, and is set to cultivate, began to develop should not be reversed after interference, etc. We are a mixture of the second and third, such as filling products to distinguish between different batches according to first, the same batch number plate layer filling products also need to separate different place (small batch samples half is placed, placed upside down), load transfer in the basket, first place 20 -- 25 ℃ to cultivate, 7 days for visual inspection, will be made into a handstand, inverted into are set, will buy 30-35 ℃ for 7 days.
6. How to determine which filling samples do not need to be cultured?
A: including damaged samples that affect the sealing performance; Failure of integrity test (leak test) samples; Intermediate control test samples that are clearly specified in the document to be discarded; Samples automatically removed by the equipment; It is clearly stipulated in the document that samples should be removed manually after each interference (the provisions should be detailed and clear and need to prove their operability and stability credibility). In addition, the removed samples should be recorded to indicate the reason for the discard.
7. Is it necessary to conduct video recording in the filling test of medium? How long does it take to keep the video?
A: not necessarily. Video is an optional auxiliary means of investigation and training. If video recording is used, it will only be used as information within the company (such as internal audit reports) and will not be required to present it to inspectors. The company shall specify in the documents the circumstances under which the video shall be recorded, the storage time of the video, etc.
8. Can the power failure be simulated in the media simulated filling test?
A: no. After power failure, AB level in the core filling area cannot guarantee asepsis. If it can be simulated, what should A level monitor do? Once grade A is out of power and there is no laminar flow protection, the aseptic state is destroyed. Even if your aseptic test is qualified, after all, the test is only representative and has its limitation of aseptic testing. The simulated power failure in the media simulation filling test cannot be used as the excuse for product release behavior. It is also possible to consider the quick power transfer within A UPS or 22S.