Practical technical manuals for prepackaged medium plates for finished products -- individualized service of Petri dishes
Personalized service is to break the traditional passive service mode, make full use of various resources advantages, and actively carry out the full range of service to meet the customer's personalized needs. We know that for antibiotic enterprises and some pharmaceutical manufacturers, it is not comprehensive to monitor the microbes in the clean area only according to the two medium (TSA and SDA) recommended by the national standard, and the suitable neutralizer is added when necessary, and the medium plate is a product customized according to the customer needs.
At present, there are two risks in the monitoring of environmental microorganism in pharmaceutical enterprises.
One of the risks is the inhibition or killing effect of API on individual microorganisms in different degrees. In particular, antibiotic production enterprises, the clean room is inevitably mixed with a certain amount of antibiotic dust, and the medium plate used in environmental monitoring can not effectively eliminate the antibiotic dust for the survival and survival of microorganisms. Environmental impact factors may result in "false negative" results in environmental monitoring of clean areas. Therefore, to restore the actual condition of the microorganism in the clean room, it is necessary to eliminate the effect of antibiotics on the microorganism. The method currently used is to hydrolyze the antibiotics that fall on the plate of the culture base. For example, beta lactam antibiotics are added to beta lactamase (penicinase), cephalosporins are added to cephalosporins, and metal enzymes are needed for Penan antibiotics.
How to add enzymes in the culture of base plates to eliminate the residues of antibiotics is mainly based on the three appendix 1105 Table 2 of the 2015 edition of the Chinese Pharmacopoeia: neutralization or inactivation method corresponding to "neutralizer or inactivation method of common interferents", and then according to the actual situation of antibiotic production enterprises, collected in its production The amount of antibiotic dust in the environment was finally determined by adding enzyme experiments. In order to ensure the homogeneity of the added enzyme, such as TSA, SDA and so on, the medium must be added to the enzyme at the lowest temperature before the curing of the liquid, and it must be stirred at a constant speed to make the enzyme and the medium fully mixed and evenly mixed in the empty culture dish, which is doomed to be an industrialized production. Process. Of course, confirming the amount of enzyme added in each dish is the data that must be verified through validation.
Antibiotic hydrolase is a biological enzyme, and its essence is protein. Enzyme activity is closely related to temperature. The maximum temperature of enzyme activity is close to human body temperature, about 37 degrees Celsius. For example, the stability of cephalosporins is as follows:
(a production of cephalosporins)
Assessment of environmental monitoring points in clean area
Pharmaceutical enterprises should set up monitoring points through risk assessment when choosing clean zone environmental monitoring points. For example, the position of aseptic filling needle in the process of filling antibiotic products is indeed the key area for the exposure of aseptic materials, but the monitoring points also have a large number of antibiotic dust settlement, so it is not significant to select this point as a key point to monitor bacterial settlement. It is more scientific to keep the location of the needle as a sampling point. First, the concentration of antibiotic dust is slightly lower, and the next location is often easily contaminated by microbes. Therefore, the choice of environmental monitoring points for antibiotics production should pay more attention to "risk points". This is different from the environmental monitoring during the production of non antibiotic products.
Collection of antibiotic residue in the environment of clean area
The enterprise selects several high risk positions in the monitoring point determined after the risk assessment, and puts the clean empty dish in the whole production process (first called the empty dish quality) and marks it well. After the production is finished, weigh the weight of each dish, draw the amount of powder collected in each dish and record well. Each variety is done 3 times, respectively.
Enzyme addition test
After obtaining the antibiotic residues in the monitoring area, the type of enzyme added and the amount of enzyme added were determined. According to the environmental sampling point collected up to 2-10 times the maximum residual dust volume to carry out the test, the final amount of enzyme added. According to the type of antibiotics, microbiology laboratory determines the types of enzymes that hydrolyze antibiotics. According to the content of different enzyme content, the plate was prepared by coating method, and the recovery rate was tested, and then compared with the unenzyme culture medium dish. For example, one point in the workshop was collected from 3.8mg/ dish, and the test tube was dissolved with 15ml water to dissolve 3.8mg dust, then added enzyme to neutralize, then added microorganism. If the microbial growth test is acceptable, it is possible to determine the amount of enzyme needed to neutralize. It is important to note that the selected strains must be sensitive bacteria and common environmental bacteria in the antimicrobial spectrum of the antibiotic.
The two of the risk is the use of disinfectants in the clean environment of pharmaceutical companies every day. How to determine the residual disinfectant does not affect the growth of microbes and ensure that the results of microbiological monitoring in the clean area do not appear "false negative". Therefore, choosing the right culture medium can maximize the measurement of the microbial level on the surface of the clean room. Usually, the combination of lecithin and Twain -80 can eliminate the effects of quaternary amine compounds (benzalkonium bromide, benzalkonium chloride and other) disinfectants. The combination of lecithin, Twain -80 and L- histidine can eliminate the effects of aldehydes (formaldehyde, amyl two, etc.) and phenols (phenol, Resorcinol, etc.); sodium thiosulfate can be eliminated. The effects of halogen disinfectants, such as iodophor, iodine, etc.
At present, most of the culture medium used for the surface sampling of clean rooms in the environmental microbiological monitoring of foreign pharmaceutical companies is lecithin Twain tryptone soybean culture medium (TSAWLP). The selection of such a medium can effectively degrade the residues of disinfectants (mainly Ji Anlei compound disinfectants) that may remain on the cleanroom surface. But at present, it is unscientific to use the lecithin Twain tryptone soybean culture medium (TSAWLP) to monitor the surface microorganism level of the clean room when most pharmaceutical companies do not know the species and residual amount of the disinfectant in the clean room. Because the lecithin + Twain -80 contained in the lecithin Twain tryptone soybean medium is quantitative, which means that the residue of the degrading disinfectant must be within a specified limit. Therefore, the residual limit of quaternary ammonium disinfectants used is not blindly adopted, and the false negative results will also occur.
In a word, when designing a microbiological monitoring scheme for a clean room, the above two risks should be taken into consideration, which can truly restore the true level of the possible microorganisms in the production environment of the pharmaceutical enterprise.
Each production enterprise is a personalized individual, so as an environment monitoring and training base plate production enterprises, to make their own products to reflect the customer's personality, we must understand the "personality" of the customer. Through the reconsideration and design of production and construction, business processes and other modules, we will create high-quality personalized services for our customers.