Radiopharmaceutical Production

QC Testing

Bacterial Endotoxins

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 Bacterial Endotoxins

• All the pharmacopeia require that Contents

radiopharmaceuticals intended for intravenous • Acceptance Criteria
administration must be tested to ensure that the • Discussion
pyrogen concentration is within acceptable • Example Procedure
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).

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 Acceptance Criteria
Radiopharmaceutical
Production

QC Testing Acceptance Criteria: Not More Than 175 EU in the total

administered dose. The total administered dose is the maximum
Baterial Endotoxin
administered volume at expiration stated in milliliters. This is
Contents often written as 175EU/V. This test should be completed on
Acceptance Criteria every batch. The batch may be released prior to completion of
Discussion
Example Procedure 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.

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 Discussion
Radiopharmaceutical
Production
Discussion: The gel-clot test entails typical incubation period of
QC Testing 60 minutes, which is much too long to wait for a 110 min half life
18
F isotope. Consequently, product may be released for patient
Baterial Endotoxin use prior to completion of this 60 minute test. However, it is
Contents possible to perform an ‘in-process’ LAL test with incubation
Acceptance Criteria period of only 20 minute or less, and should be performed. In
Discussion
Example Procedure 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
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useful for trending endotoxin levels.
 Bacterial Endotoxins Procedure
Radiopharmaceutical
Production The 20 minute Endosafe PTS method

Methodology: The Endosafe PTS method involves a

QC Testing prepared, pre-calibrated cartridge which is loaded with all
necessary reagents. The cartridge is inserted into the PTS
Baterial Endotoxin system, loaded with the radiotracer, and the test is
performed in less than 20 minutes. The tracer may need to
Contents
Acceptance Criteria
be diluted with buffer or LAL water. In general, for each
Discussion tracer, a 1:20 dilution is required.
Example Procedure
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 ºC. 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.

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