N,N,N',N',N'',N''-hexamethyl-1,3,5-triazine-1,3,5(2H,4H,6H)-tripropanamine

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Jan 23 to 29, 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

N,N,N',N',N'',N''-hexamethyl-1,3,5-triazine-1,3,5(2H,4H,6H)-tripropanamine

EC Number:

240-004-1

EC Name:

N,N,N',N',N'',N''-hexamethyl-1,3,5-triazine-1,3,5(2H,4H,6H)-tripropanamine

Cas Number:

15875-13-5

Molecular formula:

C18H42N6

IUPAC Name:

(3-{3,5-bis[3-(dimethylamino)propyl]-1,3,5-triazinan-1-yl}propyl)dimethylamine

Test material form:

liquid: viscous

Details on test material:

Batch No.: 1166046
Purity 98%

In vivo test system

Test animals

Species:

mouse

Strain:

Balb/c

Sex:

female

Study design: in vivo (LLNA)

Vehicle:

other: DAE 433 - mixture of 40% dimethylacetamide, 30% acetone and 30% ethanol

Concentration:

5%, 10%,25% in DAE433

No. of animals per dose:

5

Details on study design:

The test substance was administered in the form of suspension in DAE 433.
The volume of the application form was constant at all groups of animals - 25 l of the appropriate dilution to the dorsum of each ear once a day morning for 3 consecutive days. The application was performed very slowly by micropipette.
Day 1:
Open application of 25μL (in the morning, by pipette) of appropriate suspensions of the test substance, the vehicle alone or the positive control to the dorsum of each ear.
Days 2 and 3:
The application procedure repeated as carried out on day 1.
Days 4 and 5:
No treatment.
Day 6:
The weight of animals was recorded. Injection 250 μL of phosphate-buffered saline (PBS) containing 7.26 x105 Bq of 3H-methyl thymidine into all test and control mice via the tail vein.

Positive control substance(s):

other: Dinitrochlorobenzene

Statistics:

For statistical calculations the software Statgraphic ® Centurion (version XV, USA) was used. Statistical evaluation of measured parameters was performed at first by applying the non-parametric Kruskal-Wallis test for the comparison of the measured effect in all treatment groups with the vehicle control group, as global test, and then the non-parametric two-group Mann-Whitney rank test (probability level 0.05) for all two-group comparisons.

Results and discussion

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Value:

1

Test group / Remarks:

Negative control

Results 2

Parameter:

SI

Value:

9.67

Test group / Remarks:

Positive Controls

Results 3

Parameter:

SI

Value:

1.38

Test group / Remarks:

25% group

Results 4

Parameter:

SI

Value:

1.03

Test group / Remarks:

10% group

Results 5

Parameter:

SI

Value:

1.01

Test group / Remarks:

5% group

Results 6

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: Negative Control Group = 250 Positive control Group = 2418 25% solution group = 345.89 10% solution group = 257.53 5% solution group = 252.88

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the given test conditions, the test substance does not elicit a sensitising response in LLNA assay.

Executive summary:

The positive control substance DNCB produced positive LLNA response at an exposure level expected to give an increase in the Stimulation Index SI ≥ 3 over the negative control group, which was in congruence with the expected mode of action of a contact allergen. The positive control also elicited a reaction pattern with statistically significant increase in ear weight. The negative control did not reveal any changes.These results demonstrate that the method performed in conditions of our laboratory has sufficient reliability.

The value of SI for the test groups treated by the test substance at all three dose levels is below the threshold, stimulation index

(SI) is < 3. The comparison of the Stimulation Indexes between the treated groups and the control group revealed that the test substance did not cause a statistically significant increase in radioisotope incorporation into the DNA of dividing lymphocytes. So the result of LLNA assay is negative. 

The animals exposed to the test substance showed no significant skin reactions and clinical symptoms of intoxication throughout the experiment.

The test substance showed a tendency to increase ear weight with relation to the dose.Statistically significant increase of ear weight was recorded in animals at the highest and the middle dose level (25%, 10% test substance concentrations).

This weight increase is caused by irritation effect of the test substance. The result showed the skin irritation potential considered as positive – it means the test substance is skin irritant.