Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
data is from peer reviewed journals

Data source

Reference
Reference Type:
publication
Title:
Local lymph node data for evaluation of skin sensitization
Author:
Gerberick et al.
Year:
2005
Bibliographic source:
Dermatitis

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Principles of method if other than guideline:
The skin sensitizing potential of the test chemical was assessed by using the Mouse Local Lymphnode Assay.
GLP compliance:
not specified
Type of study:
mouse local lymph node assay (LLNA)

Test material

Constituent 1
Chemical structure
Reference substance name:
2,2'-iminodi(ethylamine)
EC Number:
203-865-4
EC Name:
2,2'-iminodi(ethylamine)
Cas Number:
111-40-0
Molecular formula:
C4H13N3
IUPAC Name:
1,2-ethanediamine, n-(2-aminoethyl)-
Test material form:
liquid
Details on test material:
Name of the test chemical: Diethylene triamine
Molecular Formula: C4H13N3
Molecular Weight: 103.168 g/mol
Substance Type: Organic
Physical State: Liquid

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
Source: No data
Age at study initiation: 7-12 weeks

ENVIRONMENTAL CONDITIONS: No data

Study design: in vivo (LLNA)

Vehicle:
acetone/olive oil (4:1 v/v)
Concentration:
25 µl of 10,20% test chemical in acetone and olive oil
No. of animals per dose:
No data available
Details on study design:
Treatment was performed daily for 3 consecutive days. Five days after initiation of exposure all mice were injected via the tail vein with 250µl of PBS containing 20µCi of tritiatied thymidine. The mice were sacrificed 5 hours later, and draining the auricular lymph nodes were excised and pooled for each experimental group or each individual animal. The incorporation of tritiated thymidine measured by β-scintillation counting and was reported in disintegrations /minute. An SI was calculated for each chemical group as the ratio of disintegrations/minute of the treated group to the disintegrations/minute of the concurrent vehicle control group. A substance was classified skin sensitizer , if at one or more than one concentrations, it induced a three-fold or greater increase in local lymph node proliferative activity when treated with the concurrent vehicle treated controls (SI ≥3)

The approach to estimation of the relative skin sensitization potential is based on the mathematical estimation of the concentration of chemical necessary to obtain a threshold positive response (SI = 3); this is termed as the EC3 value
Positive control substance(s):
not specified
Statistics:

The approach to estimation of the relative skin sensitization potential is based on the mathematical estimation of the concentration of chemical necessary to obtain a threshold positive response (SI = 3); this is termed as the EC3 value

Results and discussion

In vivo (LLNA)

Results
Parameter:
EC3
Value:
5.8
Test group / Remarks:
test group
Remarks on result:
other: Moderate
Cellular proliferation data / Observations:
The EC3 of the test chemical was calculated to be 5.8

Any other information on results incl. tables

Table 2: Results of the LLNA study

CAS

Vehicle

LLNA%

LLNA%

LLNA%

LLNA%

LLNA%

LLNA SI

LLNA SI

LLNA SI

LLNA SI

LLNA SI

LLNA EC3

Relative Potency

111-40-0

AOO

10

25

-

-

-

6.4

12.1

-

-

-

5.8

Moderate Sensitizer

Where AOO – Acetic acid in olive oil(4:1) ; SI - Stimulation Index; 

 

Applicant's summary and conclusion

Interpretation of results:
Category 1 (skin sensitising) based on GHS criteria
Conclusions:
The EC3 of the test chemical was calculated to be 5.8.
Based on the EC3 value, the test chemical was considered to be sensitizing to mice skin
Executive summary:

The skin sensitizing potential of the test chemical was assessed by using the Mouse Local Lymphnode Assay.

 

The LLNA was conducted on groups of CBA mice (7-12 weeks of age) by mean of topical application of chemical on the dorsum of both ears at a dose of 25µl or to an equal volume of relevant vehicle (Acetic acid in olive oil (4:1))only. Treatment was performed daily for 3 consecutive days. Five days after initiation of exposure all mice were injected via the tail vein with 250µl of PBS containing 20µCi of tritiatied thymidine. The mice were sacrificed 5 hours later, and draining the auricular lymph nodes were excised and pooled for each experimental group or each individual animal. The incorporation of tritiated thymidine measured by β-scintillation counting and was reported in disintegrations /minute. An SI was calculated for each chemical group as the ratio of disintegrations/minute of the treated group to the disintegrations/minute of the concurrent vehicle control group.

A substance was classified skin sensitizer, if at one or more than one concentrations, it induced a three-fold or greater increase in local lymph node proliferative activity when treated with the concurrent vehicle treated controls (SI ≥3) The approach to estimation of the relative skin sensitization potential is based on the mathematical estimation of the concentration of chemical necessary to obtain a threshold positive response (SI = 3); this is termed as the EC3 value.

 

The EC3 of the test chemical was calculated to be 5.8.

Based on the EC3 value, the test chemical was considered to be sensitizing to mice skin