1H-1,4,7-Triazonine, octahydro-1,4,7-trimethyl-

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 03 March 2008 to 16 April 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

24 April 2002

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

Octahydro-1,4,7-trimethyl-1H-1,4,7-triazonine dihydrochlorid

Molecular formula:

C9H21N3, 2 HCl

IUPAC Name:

Octahydro-1,4,7-trimethyl-1H-1,4,7-triazonine dihydrochlorid

impurity 1

Reference substance name:

Water

EC Number:

231-791-2

EC Name:

Water

Cas Number:

7732-18-5

Molecular formula:

H2O

IUPAC Name:

Oxidane

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: SAJ 07-065
- Expiration date of the lot/batch: 23 October 2009
- Purity test date:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at 2-8 °C
- Stability under test conditions: in water, saline, propylene glycol, and carboxymethylcellulose >1 day at room temperature, > 2 days in the refrigerator, >3 days in the freezer
- Solubility and stability of the test substance in the solvent/vehicle: the highest concentration which can be used was 25% solution in ethanol:deionised water (70/30)
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: -

In vivo test system

Test animals

Species:

mouse

Strain:

CBA/Ca

Remarks:

OlaHsd

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan Netherlands - B.V. Postbus 6174 - NL 5960 AD Horst
- Females (if applicable) nulliparous and non-pregnant: yes
- Microbiological status of animals, when known: -
- Age at study initiation: 8-10 weeks
- Weight at study initiation: from 18.7g to 21.5g
- Housing: single, in Makrolon Type 1 cages with wire mesh top, and granulated soft wood bedding
- Diet: pelleted standard diet ad libitum (Harlan Winkelmann GmbH, D-33178 Borchen)
- Water: tap water, ad libitum (Gemeindewerke, D-64380 Rossdorf)
- Acclimation period: at least 5 days under test conditions
- Indication of any skin lesions: no

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3°C
- Humidity: 30-70%
- Air changes (per hr): not available
- Photoperiod: 12 hrs dark / 12 hrs light
- IN-LIFE DATES: not available

Study design: in vivo (LLNA)

Vehicle:

other: ethanol:deionised water (70/30)

Concentration:

5,10 and 25%

No. of animals per dose:

4

Details on study design:

PRE-SCREEN TESTS:
- Compound solubility: the highest concentration which can be technically used was 25% solution ins ethanol:deionised water (70/30)
- Irritation: no sign of irritation was observed up to 25%
- Systemic toxicity: none
- Ear thickness measurements: not measure
- Erythema scores:

MAIN STUDY

ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: random
- Criteria used to consider a positive response:
* exposure to at least one concentration of test item results in an incorporation of 3HTdR at least 3-fold or greater than that recorded in control mice, as indicated yb the stimulation index.
* the data are compatible with a conventional dose response, although allowance must be made for either local toxicity or immunological suppression.

TREATMENT PREPARATION AND ADMINISTRATION:
The solutions were made freshly before each dosing occasion.

Positive control substance(s):

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

Results and discussion

Positive control results:

5%: SI = 1.78
10%: SI = 1.84
25%: SI = 4.87
EC3 = 15.7% (w/v)

In vivo (LLNA)

Results

Results 1

Key result

Parameter:

SI

Value:

1.45

Test group / Remarks:

5%

Results 2

Key result

Parameter:

SI

Value:

1.84

Test group / Remarks:

10%

Results 3

Key result

Parameter:

SI

Value:

1.42

Test group / Remarks:

25%

Cellular proliferation data / Observations:

CELLULAR PROLIFERATION DATA
See table

DETAILS ON STIMULATION INDEX CALCULATION
See table

EC3 CALCULATION: not determined since all SI are below 3.

CLINICAL OBSERVATIONS: none

BODY WEIGHTS: the BW recorded prior to the first application and prior to treatment witth 3HTdR was within the range commonly recorded for animals of this strain and age

Any other information on results incl. tables

Results:

Test item concentration % (w/v)	Group	DPM	DPM-BG	number of lymph nodes	DPM per lymph node	SI
	BG I	48
	BG II	25
0 (Control)	1	2865	2829	8	353.6
5	2	4150	4114	8	514.2	1.45
10	3	5251	5215	8	651.8	1.84
25	4	4060	4024	8	502.9	1.42

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test item was found to be not a skin sensitiser under the test conditions.

Executive summary:

In a dermal sensitization study with TACN dichloride at 5, 10 and 25% in ethanol:deionised water (70/30), young adult female mice (4/group) were tested using the LLNA method according to OECD 429 guideline. No clinical signs (systemic or local) nor mortality were reported.                                            

In this study, TACN dichloride is not a dermal sensitizer according to CLP criteria.