The ASTM Fa method is the standardized procedure used to conduct the bacteria retention test (bacterial challenge test) and determines the filter’s. accordance with ASTM Standard. F Following that standard, the organism is cultured in saline .. tration, ASTM Standard F,. (ASTM. ASTM-F Standard Test Method for Determining Bacterial Retention of ASTM-F › Historical Revision Information ASTM-F
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The bacterial retention test is a parameter of filter validation that has to be evaluated accordung to the requirements of the PDA 26 report and the guidance of the FDA. The filter is mounted f8388-3 a specified device and a defined bacteria solvent is pushed through the filter.
The aim is to determine how many log-levels of bacteria the filter can reduce.
You need certain materials to conduct the test. Furthermore, a testing device with a pressure connection, pressure vessel, three valves and the test filter housing must be available see figure, simplified representation.
Moreover, buffer solutions and nutrient media like for example SLB — saline lactose broth — or TSB – tryptic soy broth must be provided. The laboratory asgm also have a sterile workbench as well as an autoclave and an incubator. First, a microbial strain from the ATCC culture needs to be cultivated. The suspension created this way can then be used for the test, but f883-83 usable only for a maximum of eight hours stored in the aastm. The cultivation of B. Bacteria of this size are considered to be very small and should be retained by a sterile filter with a pore size of 0.
The bacterial retention test according to ASTM Fa
At first the cells are counted under the microscope. For this purpose, different dilutions asrm -3 — 10 -5 of bacterial suspension have to be prepared and plated as a defined quantity 0. After incubating the plates for 48 hours, the grown colonies are counted and the viable cell concentration is calculated. The entire device must be prepared according to the defined requirements explained in the following paragraph to perform the test method.
Since the test is conducted with B. For this purpose the filter to be tested must be mounted in the filter housing of the device and then be sterilized for example in the autoclave.
This also applies to all tubes and connecting pieces. Asym, the testing device has to be assembled under the sterile workbench. For the test d838-83, a negative control has to be prepared in advance.
Subsequently, the filter to be tested is flooded after opening the first valve and the excess of compressed air is released. After that, the real test can be performed. In contrast to the negative control, the filtrated solution is transferred to a second pipe by opening another valve and wets the control filter located there.
This filter is mounted on an agar plate for cultivating possible bacteria as well and is incubated for up to 7 days. The PDA 26 report additionally includes a positive control. During this, instead of the sterile filter to be tested, a filter with a pore size of 0. After the incubation period, the negative control, as well as the sample, has to be checked for the growth of bacterial colonies.
If bacterial growth is detected on the sample, it needs to be determined which bacteria it is. This is done with Gram staining that has to be evaluated microscopically.
The test bacteria B. Accordingly, colonies of a different color for example light red or Gram-positive bacteria mean that there has been a contamination by external germs.
The bacteria retention rate can be calculated now, using the found number of colonies. If other bacterial strains are found on the nutrient medium of the sample, the test has to be declared invalid.
Filter validation: The bacterial retention test according to ASTM F838-15-a
The same applies if bacteria colonies on the negative control are found. Finally, it can be concluded that the standardized performance of the bacterial retention test bacterial challenge test according to the ASTM Fa method during filter validation, guarantees the receipt of reliable results independent of the contract laboratory conducting the test. Janet Thode Trainings – Dr.
Janet Thode Michael Thode.
What is the ISO ? What is method validation? Verification of compendial methods Method transfers – good to know Procedure and requirements for method transfers Planning phase — Preparing a method transfer Types of transfer and transfer strategy Content of a transfer plan Acceptance criteria of comparative method transfers Filter validation: Good to know Filter validation: Aspects to be considered Filter validation: Information from the filter supplier Filter validation: Product-specific studies that need to be performed by the filter user Filter integrity tests — principles and influencing variables The bacterial retention test according to ASTM V838-83 HPLC troubleshooting and method optimization D838-83 for technical scientific documents.
Necessary materials You need certain materials to conduct the test. Preparing the device The entire device must be prepared according to the defined requirements explained in the following paragraph to perform the test method. Conducting the test For the test performance, a negative control has to be prepared in advance. Evaluating the test After the incubation period, the negative control, as well as the sample, has to be checked for the growth of bacterial colonies.