2,2'-[[4-[(2-hydroxyethyl)amino]-3-nitrophenyl]imino]bisethanol

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2005

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study performed according to OECD guideline 429 in compliance to GLP.

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Reference 2

Reference Type:

other: Published secondary source

Title:

Unnamed

Year:

2006

Report date:

2006

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:

Not applicable

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

2,2'-[[4-[(2-hydroxyethyl)amino]-3-nitrophenyl]imino]bisethanol

EC Number:

251-410-3

EC Name:

2,2'-[[4-[(2-hydroxyethyl)amino]-3-nitrophenyl]imino]bisethanol

Cas Number:

33229-34-4

Molecular formula:

C12H19N3O5

IUPAC Name:

2-({4-[bis(2-hydroxyethyl)amino]-2-nitrophenyl}amino)ethan-1-ol

Details on test material:

Test item : 2-[(4-N,N-bis-(2'-hydroxyethyl)amino-2-nitrophenyl)amino]ethanol
EC number : 252-410-3
Batch number : 114B5
Content : 98.7%

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Study design: in vivo (LLNA)

Vehicle:

dimethylformamide

Concentration:

25uL of 0.5, 1.0, 2.5, 5 and 10% in dimethylformamide.

No. of animals per dose:

4 females per dose

Details on study design:

Range Finding Test : No information available

Main Study
In two independent experients, 25uL HC Blue No2 in dimethylformamide was applied to the dorsal surface of both ears at the followinh concentrations : 0.5%, 1.0%, 2.5%, 5.0%, 10.0%. A positive control group received 25% alpha-hexylcinnamicaldehyde (HCA) in dimethylformamide and the negative control the vehicle alone.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

Positive control results:

Positive control induced a positive response. The stimulation indices were 9.97 and 5.70 at the concentration of 25%.

In vivo (LLNA)

Results

Parameter:

SI

Remarks on result:

other: see Remark

Remarks:

In each experiment, during the induction phase, the test item, vehicle or positive control was applied over the ears (25uL per ear) for 3 conecutive days. After 2 days of resting, the proliferation of lymphocytes in the lymph node draining the application site was measured by incorporation of tritiated thymidine (day 6). The obtained values were used to calculate stimulation indices (SI). In the first experiment, an increase in stimulation index was observed at 5% only. In the second experiment, a dose related increase in the stimulation index was noted at concentrations between 1 and 10%, but the threshold positove value of 3 was not reached at any of the tested concentrations.

Any other information on results incl. tables

Experiment 1

Treatment	Concentration (%)	Signs of Local Irritation	Stimulation Index (SI)
B37	0.5	no	1.37
B37	1	no	1.03
B37	2.5	no	1.30
B37	5	no	3.92
B37	10	no	1.55
HCA	25	-	9.97

Experiment 2

Treatment	Concentration (%)	Signs of Local Irritation	Stimulation Index (SI)
B37	0.5	no	1.60
B37	1.0	no	0.90
B37	2.5	no	1.35
B37	5	no	1.50
B37	10	no	2.01
HCA	25	-	5.70

Applicant's summary and conclusion

Interpretation of results:

sensitising

Remarks:

Migrated information

Conclusions:

While the data is equivocal, the test substance HC Blue No 2 is considered to have a skin sensitising potential.

Executive summary:

The potential of HC Blue no 2 to induce contact hypersensitivity was examined using the local lymph node assay (OECD guideline 429). Groups of 4 female mice in each were treated with 0.5%, 1%, 2.5%, 5% and 10% of the test substance in vehicle (dimethylformamide) for 3 daily treatments. After 2 days of resting, the proliferation of lymphocytes in the lymph node draining the application site was measured by incorporation of tritiated methyl thymidine (day 6). While the data is equivocal, the test substance HC Blue No 2 is considered to have a skin sensitising potential.