Radiopharmaceutical Production - PowerPoint Presentation

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Radiopharmaceutical Production

QC TestingBacterial Endotoxins

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Bacterial Endotoxins

All the pharmacopeia require that radiopharmaceuticals intended for intravenous administration must be tested to

ensure that the pyrogen concentration is within acceptable limits.Pyrogens most often originate from gram-negative bacterial cell walls – referred to as bacterial

endotoxin

and that are

readily detected by a gel-clot or other techniques based on Limulus amebocyte lysate (LAL).

ContentsAcceptance CriteriaDiscussionExample Procedure

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Acceptance Criteria

Acceptance Criteria:

Not More Than 175 EU in the total administered dose. The total administered dose is the maximum administered volume at expiration stated in milliliters. This is often written as 175EU/V. This test should be completed on every batch. The batch may be released prior to completion of the test but should the dose should not be injected into a patient until the batch has passed this test.

Procedure:

There are widely used and acceptable tests for assessing presence of bacterial

endotoxin in a radiopharmaceutical preparation. One is the gel-clot technique using Limulus Amebocyte Lysate (LAL). The bacterial endotoxin

test can also be performed with devices that utilize the turbidity and kinetic measurement of gel formation. It is essential that the test is validated for potential inhibition (and hence false negative result) and positive controls. Slide4

Discussion

Discussion:

The gel-clot test entails typical incubation period of 60 minutes, which is much too long to wait for a 110 min half life 18F isotope. Consequently, product may be released for patient use prior to completion of this 60 minute test. However, it is possible to perform an ‘in-process’ LAL test with incubation period of only 20 minute or less, and should be performed. In addition to the gel-clot method, two other methods: turbidimetric and kinetic are possible alternate that can be considered. A full 60 minute test may be performed at a specified time post-release if required. It is recommended that the shorter version LAL test is validated for its applicability. USP specifies that the product can be distributed under control after the bacterial

endotoxin

test is initiated. However,

endotoxin test results should meet the acceptance criteria before administrating the product to humans. The

Chromogenic test is demonstrated here. This test is complete in 20 minutes and gives a quantitative value for the concentration of the endotoxins in the sample. This is very useful for trending endotoxin levels.Slide5

Bacterial

Endotoxins ProcedureThe 20 minute

Endosafe PTS method Methodology: The Endosafe PTS method involves a prepared, pre-calibrated cartridge which is loaded with all necessary reagents. The cartridge is inserted into the PTS system, loaded with the radiotracer, and the test is performed in less than 20 minutes. The tracer may need to be diluted with buffer or LAL water. In general, for each tracer, a 1:20 dilution is required.

Performing a Routine Test: Press the MENU key on the PTS keypad to turn the unit on. The unit will perform a self test and heat itself to 37

º

C. This will take approximately 5 minutes. The unit will display "SELF TEST OK" then "INSERT CARTRIDGE.“ Remove a cartridge from its packaging and insert it into the unit. The unit will heat the cartridge to 37

ºC. Dispense the sample: With (4) separate pipette tips, place 25 µL of sample into each of the four wells. Press ENTER on the keypad to start the test.

Test Results: When the test is complete, the PTS reader gives an audible signal and displays the results. Link to DemonstrationSlide6

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