1,3-Benzenediol, 4,4',4''-(1,3,5-triazine-2,4,6-triyl)tris-

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation

Remarks:

in vivo

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

2001-01-09 to 2001-02-22

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: OECD study, GLP, read across substance

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

LLNA incorporated adopted on 17-Jul-1992

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:

-

EC Number:

444-500-0

EC Name:

-

IUPAC Name:

444-500-0

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan Netherlands, B.V. Postbus 6174, NL-5960 AD Horst / The Netherlands
- Age at study initiation: 7-12 weeks (beginning of acclimatization)
- Weight at study initiation: 17.5 g - 21.8g (beginning of acclimatization period)
- Housing: In groups of four in Makrolon type-3 cages with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz).
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 7 days under test conditions after health examination. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 +/- 3 °C
- Humidity: 30 - 70 %
- Air changes: 10-15h
- Photoperiod: 12 hour fluorescent light / 12 hour dark cycle with at least 8 hours music during the light period.

IN LIFE DATES:
09-Jan-2001 to 24-Jan-2001

Study design: in vivo (LLNA)

Vehicle:

other: ethanol, absolute

Concentration:

5 %, 10 % and 20 %

No. of animals per dose:

4 females

Details on study design:

TREATMENT PREPARATION AND ADMINISTRATION:
TOPICAL APPLICATION
Each test group of mice was treated by topical (epidermal) application to the dorsal surface of each ear lobe (left and right) with different test item concentrations of 5 %, 10 % and 20 % in ethanol, absolute. The application volume, 25 µl, was spread over the entire dorsal surface of each ear lobe once daily for three consecutive days. A further group of mice was treated with an equivalent volume of the vehicle alone. A hair dryer was used to dry the ear's surface as quickly as possible to avoid loss of test item applied.
ADMINISTRATION OF 3H-METHYL THYMIDINE
3H-methyl thymidine (3HTdR) was purchased from Amersham International (Amersham product code no. TRA 310; specific activity, 2 Ci/mmol; concentration, 1 mCi/ml). Five days after the first topical application, all mice were administered with 250 µl of 82.61 µCi/ml ³HTdR (equal to 20.65 µCi ³HTdR) by intravenous injection via a tail vein.
DETERMINATION OF INCORPORATED ³HTDR
Approximately five hours after treatment with ³HTdR all mice were euthanized by intraperitoneal injection of NARCOREN (Rhone Merieux GmbH, D-88471 Laupheim) at a dose of at least 2 mL/kg body weight. The draining lymph nodes were rapidly excised and pooled for each experimental group (8 nodes per group). Single cell suspensions (phosphate buffered saline) of pooled lymph node cells were prepared by gentle mechanical disaggregation through stainless steel gauze (200 µm mesh size). After washing three times with phosphate buffered saline (approx. 10 mL) the lymph node cells were resuspended in 5 % trichloroacetic acid (approx. 3 mL) and incubated at approximately +4 °C overnight for precipitation of macromolecules. The precipitates were then resuspended in 5 % trichloroacetic acid (1 mL) and transferred to glass scintillation vials with 10 mL of 'Ultima Gold' scintillation liquid and thoroughly mixed. The level of ³HTdR incorporation was then measured on a ß-scintillation counter. Similarly, background ³HTdR levels were also measured in two 1 mL-aliquots of 5 % trichloroacetic acid. The ß-scintillation counter expresses ³HTdR incorporation as the number of radioactive disintegrations per minute (DPM).

Positive control substance(s):

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

Statistics:

The mean values and standard deviations were calculated in the body weight tables.

Results and discussion

Positive control results:

ALPHA-HEXYLCINNAMALDEHYDE is regarded as a non-sensitizer at a concentration of 5 %. At concentrations of 10 % and 25 % it was found to be a sensitizer. STIMULATION INDICES of 2.4, 3.7 and 7.0 were determined with the test item at concentrations of 5 %, 10 % and 20 %

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: In this study STIMULATION INDICES of 1.0, 1.2 and 2.0 were determined with the test item at concentrations of 5 %, 10 % and 20 %, respectively, in ethanol, absolute.

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: 2489 (5 %), 2799 (10 %), 4922 (29 %) and 2443 (control group)

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU