Ashland Skin Irritation Test

Topic: Skin irritation

Test Method Number:
TM2015-02 (EU)
Short Name of TM:
Ashland RhE SIT
Year received:
2015
Responsible Organisation:
General Comments:
Similar test method to already validated and accepted ones. Not considered for follow-up

Method Description

This test is based on the in vitro test system of Reconstructed Human Epidermis (RHE), which closely mimics the biochemical and physiological properties of the upper parts of the human skin. Cells used are primary keratinocytes isolated from human skin. Irritant compounds induced reversible cell damage which can be measured after short time application (42 minutes) and long post incubation (42 hours). Cell viability in RHE models is measured by enzymatic conversion of the vital dye MTT 3-(4,5- Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. After solubilisation Thiazolyl blue is measured by optical density. Cell viability of compound is compared to negative control (DPBS) to define percent of viability (end point). Compound is classified irritant if viability is under 50%.

This test allows testing of chemical compounds to class them into 2 subgroups (irritant and non irritant) for cosmetics and pharmaceutical application.

There is an impact in...

This test is based on the in vitro test system of Reconstructed Human Epidermis (RHE), which closely mimics the biochemical and physiological properties of the upper parts of the human skin. Cells used are primary keratinocytes isolated from human skin. Irritant compounds induced reversible cell damage which can be measured after short time application (42 minutes) and long post incubation (42 hours). Cell viability in RHE models is measured by enzymatic conversion of the vital dye MTT 3-(4,5- Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. After solubilisation Thiazolyl blue is measured by optical density. Cell viability of compound is compared to negative control (DPBS) to define percent of viability (end point). Compound is classified irritant if viability is under 50%.

This test allows testing of chemical compounds to class them into 2 subgroups (irritant and non irritant) for cosmetics and pharmaceutical application.

There is an impact in terms of the Three Rs principle because this method avoids using animal to test irritant compounds. It’s an alternative method to cutaneous Draize test so it replaces animal testing.

Track Approval Status

  •  
    Submission
  •  
    Validation
  •  
    Peer-review
  •  
    Recommendation
  •  
    Regulatory acceptance/Standards
Step
Submission Show status
Validation
Peer-review
Recommendation
Regulatory acceptance/Standards