Practical Techniques of Preloading Culture Medium Plate with Finished Products
Microbial contamination is an important indicator of drug quality assessment. In order to avoid exogenous contamination from entering the drug production workshop, the 2010 version of GMP stipulates that pharmaceutical companies need to conduct dynamic or static monitoring of the clean environment, and use sedimentation bacteria flat dishes, air planktonic bacteria flat dishes, and surface contact dishes to evaluate the microbial status of aseptic production.
In the past, pharmaceutical companies generally met their needs through self-prepared culture plates, but there were some insurmountable problems due to self-prepared culture plates, such as:
The preparation of sterile flat dishes requires conditions such as matching dishes, packaging, vacuuming, and gamma ray end point sterilization. The process is complicated and difficult to achieve. If end point sterilization is not carried out, it is easy to cause false positive results;
After 100% pre-culture, sterility of the plate needs to be checked to prevent the introduction of foreign contaminants into the monitoring area.
Using containers to hold flat dishes, or directly passing flat dishes into clean areas for use, can also contaminate the production area.
Self-prepared flat dishes have a short valid period and are often prepared in small quantities, which increases batch differences and management costs.
Therefore, more and more pharmaceutical companies choose to use standardized commercial pre-filled culture flat dishes. With the in-depth implementation of the new version of GMP, self-prepared flat dishes will gradually be replaced by pre-filled culture flat dishes.
In order to strengthen the environmental monitoring and supervision of clean areas, the State Food and Drug Administration issued the "Notice on the Classification and Definition of Medium Products" on September 27, 2008, which included the medium products in the management of medical apparatus, and required the regulatory authorities and production and business units to comply with the implementation. The specific content of the classification of medium products is defined: basic, nutrition, general sterilization, transportation, preservation of medium and other media products managed as Class I medical apparatus; and drug susceptibility testing, identification, special/selection sterilization, isolation, selection of medium as Class II medical apparatus management products.
On November 26, 2013, the State Food and Drug Administration classified the culture media products in the "6840 In Vitro Diagnostic Reagents Classification Subcatalog (2013 Edition) " in the "Notice of the Food and Drug Administration on Printing and Distributing the Classification Subcatalog of In Vitro Diagnostic Reagents".
On September 5, 2014, the State Food and Drug Administration stated in the fourth point of the "Notice on the Implementation of Quality Management Standards for the Production of Medical Apparatus" that "since January 1, 2018, all medical apparatus manufacturers shall comply with the requirements of the production quality management standards for medical apparatus". The regulations clearly stipulate the production of culture media.
In order to help the majority of users understand and use commercial pre-filled culture plates, we have prepared this "Practical Technical Manual for Finished Pre-Filled Petri Plates", which is based on practice and combined with the theory of microbial control in clean areas. It explains in detail how to effectively control microorganisms in clean areas to achieve a stable and controllable level of microorganisms in clean areas, as well as the methods for testing microorganisms in clean environments and their development history. It also describes the production management and quality control of pre-packaged finished petri platters used in the process of microbial testing in clean areas. Quality requirements for pre-packaged finished petri platters, how to use the product, and common questions and answers during use.
This manual is divided into 8 chapters, introducing actual cases of microbial monitoring in clean areas, and also giving basic ideas on how to monitor the environment of clean areas. It discusses some advanced concepts abroad in detail, hoping to help everyone get effective help in the process of using finished culture media flat dishes, so that the monitoring results reflect the true level of microorganisms in clean areas. There may be some duplicate content in different chapters of the manual, but this duplicate content is very necessary, mainly to emphasize its importance.
This manual is not only suitable for the pharmaceutical industry, but also for environmental microbial testing in hospitals, CDC, food, cosmetics, microelectronics, and other industries.
Due to the fact that the industry standards for pharmaceuticals, especially biological products, are constantly being updated, coupled with the limited level of editors and the lack of time, the manual will inevitably have shortcomings. We sincerely hope that readers and colleagues will criticize and correct it.
Chapter 1: Clean Area Microbial Control Specifications and Requirements
First, the control of microorganisms in clean areas
In addition to directly reflecting the efficacy and safety of drugs, the quality of drugs is also reflected in the stability and consistency of drug quality. If the production environment is not strictly controlled during the production process of drugs, it is easy to be contaminated or cross-contaminated by microorganisms, dust particles, etc. These characteristics of drugs put forward various requirements for the indoor environment in which the drug production process is located, the temperature, humidity, cleanliness, pressure difference, discharge method and the maintenance of personnel hygiene.
In order to ensure that the quality of medicines meets the requirements, the production environment of traditional Chinese medicine products in pharmaceutical companies needs to be processed in the clean area. The clean area refers to the removal of dust particles, microorganisms and other pollutants in the air within a certain space range, and the indoor temperature and humidity, cleanliness, pressure difference, emission method, vibration noise, illuminance and static electricity are controlled within a certain range, and specially designed rooms are given. That is, through heating ventilation and air conditioning system (HVAC system) to control, in China GMP called air conditioning purification system, is a key system for pharmaceutical companies, it has an important impact on whether pharmaceutical companies can achieve their goal of providing safe and effective medicines to patients. If the drug production environment is reasonably designed, constructed, commissioned, operated and maintained, it will help to ensure the quality of the product, improve the reliability of the product, and reduce the initial investment cost and later operation cost of the enterprise.
GMP (2010 version) clean area classification
Air cleanliness refers to the degree to which the air in a clean environment contains suspended particles. The less the number of particles in the air, the higher the cleanliness. All countries in the world have corresponding cleanliness standards. The GMP (2010 version) of our country stipulates that the design of clean areas must meet the corresponding cleanliness requirements, including "static" and "dynamic" standards.
The clean areas required for the production of sterile medicines can be divided into the following four levels:
Class A: High-risk operating areas, such as filling areas, areas where stopper drums and open packaging containers in direct contact with sterile preparations, and areas where sterile assembly or joining operations are performed, should be maintained with a unidirectional flow console (hood). Unidirectional flow systems must be evenly ventilated in their working areas at a wind speed of 0.36-0/s (guidance value). There should be data to prove the status of unidirectional flow and verify it.
In a closed isolated operator or glove box, a lower wind speed can be used.
Class B: refers to the background area where the Class A clean area for high-risk operations such as aseptic preparation and filling is located.
Class C and Class D refer to clean areas for less important steps in the production of sterile pharmaceutical products.
Note:
(1) In order to confirm the level of Class A clean areas, the sampling volume of each sampling point shall not be less than 1 cubic meter. The level of airborne particles in Class A clean areas is ISO 4.8, with a limit of ≥ 5.0 μm as the standard. The level of airborne particles in Class B clean areas (static) is ISO 5, and at the same time includes suspended particles of two particle sizes in the table. For Class C clean areas (static and dynamic), the level of airborne particles is ISO 7 and ISO 8 respectively. For Class D clean areas (static), the level of airborne particles is ISO 8. The test method can be referred to ISO 14644-1.
(2) When confirming the level, a portable dust particle counter with a short sampling tube should be used to avoid sedimentation of suspended particles ≥ 5.0 μm in the long sampling tube of the remote sampling system. In a unidirectional flow system, an isokinetic sampling head should be used.
(3) Dynamic testing can be performed during routine operation and medium simulation filling to demonstrate a dynamic cleanliness level, but the medium simulation filling test requires dynamic testing under "worst-case conditions".
