Practical technical manual of finished preloaded culture medium plate 1
發(fā)布時(shí)間：
2022-12-27
作者：
Practical technical manual of finished preloaded culture medium plate 1
preface
Microbial contamination is one of the important indicators, drug quality evaluation to avoid exogenous pollution into the pharmaceutical production workshop, 2010 version of the GMP regulation: pharmaceutical companies to clean environment for dynamic or static monitoring, using settlement bacteria AGAR, air planktonic bacteria AGAR and surface contact properly evaluate aseptic production situation of microorganisms.
In the past, pharmaceutical companies generally met their use requirements by self-matching petri dishes, but there were some insurmountable problems in self-matching petri dishes, such as:
Preparation of asepsis AGAR culture medium is required with melamine, packaging, vacuum, gamma rays, terminal sterilization conditions, craft complex, difficult to achieve, if not for terminal sterilization easy to cause false positive results;
After 100% preculture, it is necessary to carry out aseptic examination on the plate culture medium to prevent foreign pollutants from entering the monitoring area.
The use of containers for placing flat dishes, or directly into the clean area for use, will also contaminate the production area;
The validity period of self - allocated flat ware is short, and it is often prepared in small quantity for many times, which increases the inter-batch difference and management cost.
As a result, more and more pharmaceutical companies choose to use standardized pre-filled commodity culture plate. With the further implementation of the new GMP, the self-leveling dish will be gradually replaced by the pre-filled culture plate.
In order to strengthen the supervision and management of clean area environment monitoring, the state food and drug administration on September 27, 2008, issued the notice on the product categories defined culture medium type, the medium products listed in the management of medical equipment, and demanded that the regulators and the production and business operation entity shall comply with them. Medium class product categories define specific content: the basic medium, nutrient medium, general enrichment medium, transport medium, storage medium as Ⅰ medium product of medical equipment management; And drug sensitive test media, differential medium, special/choice enrichment medium, separation medium, choose medium as Ⅱ class of medical equipment management of media products.
On November 26, 2013, the state food and drug administration in the food and drug supervision bureau about classification of subdirectories notice issued by the in vitro diagnostic reagent, medium products division in 6840 in vitro diagnostic reagents classification subdirectory (2013 edition).
On September 5, 2014, the state food and drug administration in "about medical equipment production and the quality control standard for the implementation of relevant issues notice" in the fourth, it is pointed out that "since January 1, 2018, all medical device production enterprise shall conform to the requirements of medical equipment production quality management standard". The regulation clearly stipulates the production of culture medium.
To help users understand and use goods pre filling culture plate, we write this on the finished product with medium AGAR practical technical manual, manual on the basis of practice, combined with clean areas of microbial control theory, a detailed expounded how to effectively control the clean area microorganisms can achieve clean area stability, can control level, and clean environment microbial detection methods and their development history, elaborated in clean areas used in the process of microbial detection with the finished product of medium plate production management and quality control, with the finished product of medium plate quality requirements, the product is how to use, Common questions and answers during use.
This manual is divided into eight chapters, the introduction of clean areas microbial monitoring process of the actual case, also gives the basic ideas of how to monitor the clean area environment, a detailed discussion on the some of the foreign advanced ideas, hoping to help people in the use of the finished product effectively with the help of the culture medium of plating process, the monitoring results reflect the true level of clean area microorganisms. There may be some repetition in different sections of the manual, but this repetition is necessary to emphasize its importance.
This manual is applicable to environmental microbiological detection in hospitals, centers for disease control, food, cosmetics, microelectronics and other industries.
Due to drugs, especially biological products industry standards have been update the status, combined with the editor level is limited, time hasty, manual unavoidably exist deficiencies, we sincerely hope that readers and comment.
Chapter 1 microbiological control regulations and requirements in clean areas
I. control of microorganisms in clean areas
1
Drug quality in addition to directly reflected in the efficacy and safety, but also on the stability of the drug quality and consistency, if does not carry on the production environment in the process of drug production strictly controlled, is vulnerable to microorganisms, such as dust particles contamination or cross-contamination. These characteristics of drugs, the indoor environment, the process of drug production of air temperature, humidity, cleanliness, differential pressure, discharge mode and personnel health keeping in various aspects such as all kinds of requirements.
Accords with a requirement in order to ensure the quality of the drugs, pharmaceutical companies in the pharmaceutical production environment needs to be done in processing in a clean area, clean area refers to the certain space within the scope of the dust particles in the air, microbial pollutants such as exclusion, and the indoor temperature and humidity, cleanliness, differential pressure, discharge way, vibration noise, intensity of illumination and electrostatic control within a certain range, and special design of the room. By heating ventilation and air conditioning system (heating ventilation and air conditioning, HVAC systems) to control, according to GMP air conditioning purification system in China, is a key system of pharmaceutical enterprises, it provides patients with pharmaceutical companies can realize its like a safe and effective drug target and its importance. If the drug production environment and reasonable design, construction, commissioning, operation and maintenance, help to ensure that the quality of the products, improve the reliability of products, at the same time reduce the enterprise cost of investment in the early and late running cost.
2
GMP (2010 edition) clean zone classification
Air cleanliness refers to clean environment in the air containing the extent of the amount of suspended particles, including the number of particles in the air is less, the higher the cleanliness, countries around the world are all in the corresponding standards of cleanliness and GMP in China (2010 edition), the design must conform to the corresponding cleanliness requirement of clean area, including reached the standard of "static" and "dynamic".
The clean areas needed for the production of aseptic drugs can be divided into the following four levels:
Class A: high-risk operations area, such as filling area, placing barrels and stopper is exposure to direct contact with sterile products packaging container operation area and aseptic assembly or connection area, ought to use uniflow work station (cover) to maintain the area's environment. The unidirectional flow system must provide uniform air supply in its working area with a wind speed of 0.36-0.54m/s (guide value). Data should be available to verify and verify the state of one-way flows.
Low wind speeds can be used in closed isolation operators or glove boxes.
Class B: refers to the background area of grade A clean area where high risk operations such as sterile preparation and filling are carried out.
Class C and class D: refers to the clean area of a sterile drug with a lower degree of importance in the production process.
Practical technical manual of finished preloaded culture medium plate 1
Note:
(1) to confirm the level of class A clean area, the sampling amount of each sampling point shall not be less than 1 cubic meter. The level of aerosol particles in class A clean area is ISO 4.8, and the range of aerosol particles in class A clean area is greater than or equal to 5.0. Class B clean (static) aerosols are of the class ISO 5 and include two aerosols in the table. For class C clean areas (static and dynamic), the aerosol levels are ISO 7 and ISO 8, respectively. For class D clean (static) aerosol particles the level is ISO 8. The test method can refer to iso14644-1.
(2) when the recognition level is confirmed, a portable dust particle counter with a shorter sampling tube should be used to avoid settling the suspended particles with a value of 5.0 for a long sampling tube of the remote sampling system. In a one-way flow system, isodynamic sampling heads should be used.
(3) dynamic testing can be in normal operation, the culture medium to simulate filling process, prove that to achieve the dynamic level of cleanliness, but medium filling simulation test requirements under the "worst situation" dynamic testing.
Requirements for microorganisms at all levels in a clean area
Dynamic monitoring of microorganisms should be carried out to assess the microbiological status of aseptic production. The monitoring methods include sedimentation bacterium method, quantitative airborne phytoplankton sampling method and surface sampling method (such as cotton swab wiping method and contact plate method). Dynamic sampling should avoid adverse effects on clean areas.
Monitoring of surfaces and operators shall be carried out after the completion of key operations. In addition to normal production operation monitoring, microbiological monitoring can be added after system verification, cleaning or disinfection, etc.
Practical technical manual of finished preloaded culture medium plate 1
Note:
(1) all values in the table are average values.
(2) the exposure time of a single sedimentation disc can be less than 4 hours, and multiple sedimentation discs can be used for continuous monitoring and accumulation counting at the same location.
4
Clean areas are classified by status
Dynamic means that the production equipment operates in the predetermined process mode and a specified number of operators operate in the field.
Static state means that all production equipment is installed and ready, but no production activity and no operator is present.
5
Clean areas are classified by airflow pattern
Clean area of air distribution for clean in production area to achieve a particular level of clean air, to limit and avoid dust particles and bacteria attached on the dust particles of product, the direct contact with the product packaging materials, equipment, containers, utensils of pollution and use of purify air flow state and distribution status.
The indoor air organization has laminar flow and turbulent flow according to the flow state, laminar flow is divided into vertical one-way flow (laminar flow) and horizontal one-way flow (laminar flow).
One-way flow refers to the flow of air in the same direction in a stable and uniform manner and at a sufficient rate. One-way flow can continuously remove particles from key operating areas, as shown in the following figure:
Practical technical manual of finished preloaded culture medium plate 1
Vertical one-way flow clean zone:
Vertical one-way flow controls pollution in all directions at a suitable airflow speed. The faster the air flows, the better the cleanliness. Generally, in the design of one-way flow cleanroom, the internal section custom v of class A vertical one-way flow is greater than 0.36m/s.
Day-to-day operations of the most common vertical uniflow device for class A laminar flow hood, norms is lesser, mainly used in high-risk areas in protection products, such as aseptic filling in packaging and sterilization after implements the preservation of open area.
Horizontal one-way flow clean area: horizontal one-way flow clean area is full of high efficiency filter on the side air supply wall, the opposite side air return wall is full of medium effect filter (or combine with return style column). Airflow through the state of high efficiency air filter to clean, and the horizontal level uniflow state to uniform through the work area, take away the dust particles of the working area, the remaining heat and wet, the return air into the return air plenum. The most common horizontal one-way flow device in daily production is the horizontal purification bench with small specifications.
Level uniflow cleanroom must consider to overcome the influence of gravity settling of dust particles, therefore, in the design of A grade level uniflow clean indoor section customs should be slightly larger than the vertical uniflow cleanroom wind speed.
Turbulence area
Turbulent flow clean room clean air through the cover after the export of high efficiency filter of air supply outlet diffusion into the interior, clean air rapidly spreading around, mixed with indoor air, dilution of indoor air pollution, and carries on the exchange with indoor heat load, after mixing with indoor airflow dust, the positive pressure of the air, from the inlet line to walk, the diagram below:
Practical technical manual of finished preloaded culture medium plate 1
There are four main airflow tissues in the turbulence cleaning room:
The air supply at the top of the hole plate is divided into the air supply at the top of the whole hole plate and the air supply at the top of the partial hole plate.
The airflow arrangement is suitable for high buildings over 4 meters.
The high efficiency filter tuyere diffuses the top to send the air, is the production USES the clean area common airflow organization form.
Side supply air. This kind of form is suitable for low - storey workshop, and is used for old workshop reconstruction. There are many eddies in the side air supply room, and the cleanliness can only reach grade C.
6
Air supply and ventilation times in the clean area
As mentioned above, the difference between turbulence clean areas or key indicators uniflow a clean area is the air flow state in a clean area, and the air flow state is determined and the flow of the air way and speed, therefore, specify the size of the space unit time clean air quantity, is crucial for cleanliness. The following table shows the relationship between the number of clean room operators, air ventilation times and indoor cleanliness.
Practical technical manual of finished preloaded culture medium plate 1
Note:
The lower limit of indoor ventilation is 30min, and the cleanliness should reach 80%.
Uniflow (laminar flow) clean area air output = clean area area (㎡) X wind speed;
Turbulent flow (non-one-way flow) the air supply in the clean area = the volume of the clean area (m3) X times of air exchange (second /h).
The number of ventilation times in the clean environment of drug production should be determined according to different dosage forms, and the regulation of ventilation times should be verified and determined according to the calculation of heat balance and air balance.
7
Sources of microbial contamination in clean areas
Microbial pollution refers to the generation and attachment of microorganisms, which bring adverse effects to a specific environment. If bacteria get the necessary nutrients, the right amount of water and the right amount of temperature and humidity, they can reproduce very quickly, very quickly, and thus harm the clean areas where they are produced. Clean area pollution can be divided into two types. The first is the pollution of inorganic substances (inanimate objects), such as suspended particles in the air and particles on materials. The second is organic pollution, such as microorganisms. There are five main sources of microbial pollution: people, equipment, materials, air and water.
(1) personnel pollution
As is known to all, people are the main source of pollution in clean areas, and human pollution in clean areas is an endless process. Skin chips, hair, microbes in the body, etc.
Bacteria/per minute
Sneeze 12-3400
Cough 1-1000
Singing 1-128
Speech 1-28
Calm breathing 0-4
Take the operator of the clean area for example:
When sitting or standing still, up to 100,000 particles (10,0000/min) travel per minute.
The number of particles is as high as 500,000 (50,0000/min) while sitting with moderate body and arm movements.
Up to 2.5 million particles (250,000 /min) during normal walking
As many as 10 million particles (1000,0000/min) during fast walking
Up to 30 million particles (3000000 /min) are installed and repaired.
There are more than 100 billion microbes living in the human body. There are up to 400 kinds of microbes, distributed in all parts of the body.
Mouth:
A milliliter of saliva contains more than 100 million bacteria, mainly e. coli, e. coli, anaerobic bacteria
Sex coccus, lactobacillus, the bacteria in the mouth can be cleared in 6 hours after the formation.
Skin:
The skin surface is changed every four days, and 4-6 grams of skin flakes are lost every day. Each skin chip contains about 4 bacteria.
There are mainly staphylococcus epidermis, staphylococcus aureus.
How to control the pollution of people?
First of all, GMP training should be carried out for personnel engaged in preparation operation and drug inspection, who should receive professional technical training and have basic theoretical knowledge and practical operational skills. Those who have special requirements for preparation and operation of preparations and drug inspectors should also receive corresponding professional technical training. All personnel engaged in preparation should be familiar with GMP standard.
Second, clean room (area) and limited to the staff in the room and the approved personnel into the clean room (area) the personnel shall not make up and wear decorations, not bare hand direct contact with drugs. Persons entering the cleansing area shall have a health record and have at least one physical examination every year. Persons suffering from infectious diseases, skin diseases or wounds on the surface of their bodies shall not engage in any work related to the production of products.
Third, clean room staff regularly train basic microbiology theories and related pollution sources to wear aseptic clothing in the correct way.
Aseptic operations, such as: speed of movement during operation, operating range and indoor airflow direction, contact of aseptic materials or articles, sampling and disinfection methods. It should also include safe operation of the clean room, cleaning and maintenance of the clean room, and personal hygiene.
(2) pollution of facilities and equipment
Facilities are rarely an inherent source of microorganisms. However, design defects, poor structural materials and improper operation can also have a significant impact on pollution levels. Equipment together with other sources constitutes a source of pollution.
The new equipment could be designed to bring the source of internal microbial contamination to a minimum level to reach a hygienic level. On the contrary, microbiological control of these devices is very difficult to solve.
In addition to the design of the equipment will bring microbial contamination, explain the equipment clean, after completion of every working procedure of the pharmaceutical production, pharmaceutical equipment for cleaning is the essential means to prevent drug contamination and cross-contamination. After the production of products, there will always remain a number of raw materials and microorganisms. Under appropriate temperature and humidity, microorganisms can reproduce in large Numbers with the organic matter in the residue as nutrition and produce various metabolic products, thus greatly increasing the complexity and harm degree of the residue. If these residual raw materials, microorganisms and their metabolites enter the next batch of production process, they will inevitably have adverse effects on the next batch of products. Therefore, these pollution sources must be removed from the circulation of drug production through cleaning.
The cleaning method of process equipment can be divided into manual cleaning method and automatic cleaning method. The manual cleaning method is characterized by manual holding of cleaning tools, cleaning equipment according to predetermined requirements, and determining the degree of cleaning according to visual measurement until the cleaning is completed. The automatic cleaning method is characterized by the automatic special equipment according to certain procedures to complete the entire cleaning process automatically.
Regardless of the cleaning method, the key is the difficult point of clean cleaning process equipment, such as: ingredients can clean the bottom of the blade surface of impeller is the most difficult to clean, the cleaning validation is to consider the water flow velocity, temperature, cleaning time, and so on factors, finally the parameters of the revised according to the results of the validation of cleaning SOP.
Practical technical manual of finished preloaded culture medium plate 1
(3) material pollution
Materials are pollution sources and one of the pollution sources we should pay attention to. Materials are the basic parts of products, especially the raw and auxiliary materials and inner packaging materials that directly contact with products. Therefore, in the key process of product, we need to control the microbial level of the material, so as to avoid the pollution of the clean area caused by the material pollution and ultimately affect the product quality.
Therefore, the materials entering the clean area, the key areas of microbial load limitation, are potential microbial pollution sources. Microorganisms can be attached on the raw material, these microbes are hard to be a normal risk assessment, so the supplier audit for the material, partition management, in accordance with material source building materials of microbial monitoring program and so on are now commonly used method to control the material contamination.
But we have to be paid more attention material again pollution in the process of storage, transfer, material must be purified into a clean area disinfection processing, appearance, before entering the clean area shall be carried out in accordance with the enterprise internal SOP purification room, material purification, disinfection, transfer line should be separated from personnel enter the route, to avoid cross contamination. It is mainly caused by the "sticky dust bacteria" caused by the transmission equipment used in the material transfer process and the air pollution caused by the air flow during the transmission process.
If the materials used in elevator transportation, people and goods should not be transported together. The elevator shall be located in an unclean area, and if it is located in a clean area, a buffer zone shall be set at the elevator entrance.
(4) air pollution
The air is clean area pollution sources is carried, natural microbial contamination in the air are, microorganism through attachment on the inert particles in the air, such as dust particles and water droplets, skin crumbs, etc., affect product, cause pollution.
The main way to control microbial contamination in pharmaceutical production is
暫無(wú)數(shù)據(jù)