Quality Control of

Radiopharmaceuticals
BIOLOGICAL TESTS

JOHN KARL MACROHON

Biological Tests
Purpose: test for the presence of microorganisms
(bacteria, fungi, etc.) should be examined for ALL
pharmaceuticals intended for human administration
 living organisms also can produce metabolic

byproducts (endotoxins) that can undesirably

affect the radiopharmaceutical preparation
Sterility Test
Biological Test

 Sterility testing - demonstrate the presence

or absence of extraneous viable
contaminating microorganisms designed for
human use
 autoclaving for long-lived
radiopharmaceuticals and membrane
filtration/direct inoculation for short-lived
ones
Sterility Test
Biological Test

A proper sterility test involves the

incubation of the radiopharmaceutical
sample for 14 days in
 Two Growth Media

1. Fluid thioglycollate medium for

growth of aerobic and anaerobic
bacteria
2. Soybean-casein digest medium for
growth of fungi and molds
Sterility Test
Biological Test

Membrane filtration
 add an inoculum of a small number of viable
microorganisms <100 CFU to the final portion of sterile
diluent used to rinse the filter.
Direct inoculation
 add an inoculum of a small number of viable
microorganisms <100 CFU to the medium
*Incubation Time: <5 days
Pyrogenicity Test
Biological Test

Pyrogens - polysaccharides or proteins

produced by the metabolism of
microorganisms (endotoxins)
 fever, chills, malaise, joint pain, sweating, headache,
and dilation of the pupils within 30 min to 2h after
administration
Pyrogenicity Test
1. Rabbit Test
 monitoring of the temperature of three healthy
rabbits for 3 h after injection of the test sample
 Pyrogenicity Test
Biological Test
2. Limulus amebocyte lysate (LAL) method
LAL extracts from horseshoe crabs
(limulus) reacts with gram-negative
bacterial endotoxins concentrations
 Gel clot - thicker the gel, the greater the
concentration of pyrogens
 Gram-negative endotoxins - most
important source of pyrogen
contamination
USP Specification: no clotting
Toxicity Test
Biological Test

Toxicity tests should be applied for ALL

radiopharmaceuticals approved for human
use
 LD50/60  toxic effect of a
radiopharmaceutical by determination of
the dose required to produce mortality of
Such test should be done for at least two
50% of a species in 60 days after different species of animals but is
administration of a radiopharmaceutical nowadays replaced with cell culturing and
computer modeling
dose
 For determination of the LD50/60, the test substance
is injected in increasing dosages into a large group
of animals. The dosage at which 50% mortality of
the animals is observed in 60 days following
administration is established as the LD50/60 for the
material. The test must be carried out in at least two
species of animals.
Toxicity Test
Biological Test
THANK YOU!