Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Performed to GLP standards.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guideline
Qualifier:
no guideline available
Principles of method if other than guideline:
The purpose of this study is to determine the eye irritation potential of trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate using the SkinEthic Reconstituted Human Corneal model after a treatment period of 10 minutes. It is thought that irritant chemicals are able to penetrate the stratum corneum and are sufficiently cytotoxic to cause cell death in the underlying cell layers. Cytotoxicity is determined by the reduction of MTT to formazan by viable cells in the trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate treated tissues relative to the negative control.
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals / tissue source

Species:
other: in vitro test
Strain:
other: in vitro test
Details on test animals or tissues and environmental conditions:
Incubation conditions: 37 C, 5% CO2 in air.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: not applicable as in vitro test.
Amount / concentration applied:
Main test: 30 mg of trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate
Negative control: 30 microlitres os solution A (Na2HPO4 0.142 g/l; Glucose 1.802 g/l; HEPES 7.149 g/l; KCl 0.224 g/l; NaCl 7.597 g/l)
Positive control: 30 microlitres of 1% w/v Sodium Dodecyl Sulphate (SDS)
Duration of treatment / exposure:
10 minutes
Number of animals or in vitro replicates:
No animals involved - in vitro test.
Details on study design:
Experimental design of this study consists of a test for direct reduction of MTT by the test materials followed by the main test. For the main test, triplicate SkinEthic tissues are treated with 30 mg of trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate for 10 minutes. Triplicate tissues treated with 30 microlitres of solution A served as the negative control and triplicate tissues treated with 30 microlitres of 1% w/v Sodium Dodecyl Sulphate served as the positive control. After the exposure period, each SkinEthic tissue was rinsed and taken for MTT loading and extraction. The absorbency of triplicate alquots of the extracted MTT solution for each SkinEthic tissue is measured and the cell viability for each is calculated.

Results and discussion

In vivo

Results
Irritation parameter:
other: % Viability
Remarks on result:
other: in vitro test: see remarks on results section below for scores
Irritant / corrosive response data:
The mean OD50 values and mean viabilities for each treatment group are given in table 1.

Any other information on results incl. tables

Table 1. Assessment of eye irritation potential - viability of RHC tissues.

Material  Mean Tissue Viability Mean OD540 % Viability
Negative Control 0.919 0.937 100
0.954
Positive Control  0.282 0.295 31.5
0.308
trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate 0.82 0.839 89.5
0.858

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: expert judgment
Conclusions:
This study suggests that trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate is considered to be a non-irritant.
Executive summary:

In a primary eye irritation study 30 mg of trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate without a vehicle was instilled onto a reconstituted human corneal epithelium model. for 10 minutes. After the exposure, the tissues were rinsed and taken for MTT testing. Irritation was scored by the method of % cell viability.

 In this study, trisodium 6,6',6"-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoate is not an eye irritant.