In 2010 it has moved back to the main body of the document.
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 preparation is done per the associated annex which defers to ISO 10993-12.
Three rabbits are used weighing at least 2 kg of either sex from a single strain.
Shaving of the fur is done 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 injection and at 24 +/-2, 48 +/-2, and 72 +/-2 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. Only the 24, 48, and 72 hour scores are used for the overall scoring calculation.
How to calculate the score is explained in some mind blowing convoluted fashion. But ultimately it is the overall mean score for both test and control. 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 preparation is done per the associated annex which defers to ISO 10993-12.
Three rabbits are used weighing at least 2 kg of either sex from a single strain.
Shaving of the fur is done 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 injection and at 24 +/-2, 48 +/-2, and 72 +/-2 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. Only the 24, 48, and 72 hour scores are used for the overall scoring calculation.
How to calculate the score is explained in some mind blowing convoluted fashion. But ultimately it is the overall mean score for both test and control. 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.
