2002 it had been moved to an informative annex.
There is an exemption statement that test articles are excluded when the pH is less than or equal to 2 or greater than or equal to 11.5 or those already known as an irritant in other tissues.
If there is to be preparation of the test material the the preparation is done per the associated annex which defers to ISO 10993-12.
Two rabbits are used weighing at least 2 kg of either sex from a single strain
The fur is shaved 4 to 18 hours before dosing. The needle size used for dosing should be the smallest appropriate for the sample viscosity. Five, 0.2 ml injections are made of each of both polar and non polar extracts and associated controls. Observations (scoring) are made immediately after dosing and at 24 hours, 48 hours, and 72 hours post-dose
The sites are graded on a 5 point scale ranging from no(0), very slight(1), well-defined(2), moderate(3), to severe(4) for both erythema and edema.
How to calculate the score can be confusing. The overall scoring requires reading the explanation on how to calculate the score which says that there are 3 grading periods, and it also requires prior knowledge of other guidelines of this test to realize that only the 24, 48, and 72 hour scores are used for overall calculation. The calculation of the final score is explained as totaling the erythema and edema scores and dividing by 12 because there are 2 animals scored 3 times for 2 observations (erythema and edema) (2 x 3 x 2).
Final scoring is a pass/fail scenario where the study passes if the difference between the test and the control is 1.0 or less. A retest condition is included but it is a vague statement which allows for a retest in 3 animals, but no real tolerances are specified.
There is an exemption statement that test articles are excluded when the pH is less than or equal to 2 or greater than or equal to 11.5 or those already known as an irritant in other tissues.
If there is to be preparation of the test material the the preparation is done per the associated annex which defers to ISO 10993-12.
Two rabbits are used weighing at least 2 kg of either sex from a single strain
The fur is shaved 4 to 18 hours before dosing. The needle size used for dosing should be the smallest appropriate for the sample viscosity. Five, 0.2 ml injections are made of each of both polar and non polar extracts and associated controls. Observations (scoring) are made immediately after dosing and at 24 hours, 48 hours, and 72 hours post-dose
The sites are graded on a 5 point scale ranging from no(0), very slight(1), well-defined(2), moderate(3), to severe(4) for both erythema and edema.
How to calculate the score can be confusing. The overall scoring requires reading the explanation on how to calculate the score which says that there are 3 grading periods, and it also requires prior knowledge of other guidelines of this test to realize that only the 24, 48, and 72 hour scores are used for overall calculation. The calculation of the final score is explained as totaling the erythema and edema scores and dividing by 12 because there are 2 animals scored 3 times for 2 observations (erythema and edema) (2 x 3 x 2).
Final scoring is a pass/fail scenario where the study passes if the difference between the test and the control is 1.0 or less. A retest condition is included but it is a vague statement which allows for a retest in 3 animals, but no real tolerances are specified.
