Reactive Orange 143

Endpoint summary

Description of key information

The test substance is considered as skin sensitiser in LLNA study.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

14 December 2011 to 06 June 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Specific details on test material used for the study:

Identification: FAT 40854/A TE
Description: Reddish-brown powder (determined at NOTOX)
Batch: TZ 5719 / BOP 02-11
Content: 46.2 % (4 main constituents)
Test substance storage: At room temperature in the dark
Stability under storage conditions: Stable
Expiry date: 01 April 2016

Species:

mouse

Strain:

CBA:J

Sex:

female

Details on test animals and environmental conditions:

- Source: Janvier, Le Genest-Saint-Isle, France
- Age at study initiation: Young adult animals (approx. 9 weeks old)
- Weight at study initiation: Body weight variation was within ± 20 % of the sex mean.
- Housing: Animals were group housed in tagged Makrolon cages.
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17.9 – 20.7
- Humidity (%): 41 – 72
- Air changes (per hr): approx. 15
- Photoperiod (hrs dark / hrs light): 12/12

Cleaning procedures in the room might have caused the temporary fluctuations above the optimal maximum level of 70 % for relative humidity. Deviations from the minimum level of temperature occurred. Based on laboratory historical data, these fluctuations were considered not to have affected the study integrity.

IN-LIFE DATES: From: 14 December 2011 - 16 January 2012

Vehicle:

dimethylformamide

Concentration:

0, 10, 25, 50 %

No. of animals per dose:

5

Positive control substance(s):

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

Statistics:

Not performed.

Positive control results:

The six-month reliability check with Alpha-hexylcinnamicaldehyde indicates that the Local Lymph Node Assay as performed at NOTOX is an appropriate model for testing for contact hypersensitivity. See attached document 'Reliability check'.

Parameter:

SI

Value:

11.7

Test group / Remarks:

Group 2 (10 % test substance)

Parameter:

SI

Value:

13.4

Test group / Remarks:

Group 3 (25 % test substance)

Parameter:

SI

Value:

20.1

Test group / Remarks:

Group 4 (50 % test substance)

Cellular proliferation data / Observations:

All auricular lymph nodes of the animals of the experimental groups were considered larger in size compared to normal. The largest auricular lymph nodes were found in the higher dose groups. No macroscopic abnormalities of the surrounding area were noted in any of the animals.

Results Pre-screen test:

Variations in ear thickness during the observation period were less than 25 % from Day 1 pre-dose values. Red test substance remnants prevented scoring for erythema, except for scoring prior to dosing on Day 2. At that observation no erythema was noted in all animals at 25 and 50 %. Based on these results, the highest test substance concentration selected for the main study was a 50 % concentration.

 

Other results - main study:

 

Skin reactions / Irritation:

Red test substance remnants prevented scoring for erythema.

 

Systemic toxicity/Body weights:

No mortality occurred and no clinical signs of systemic toxicity were observed in the animals of the main study. Body weights and body weight gain of experimental animals remained in the same range as controls over the study period.

 

Macroscopy of the auricular lymph nodes and surrounding area:

All auricular lymph nodes of the animals of the experimental groups were considered larger in size compared to normal. The largest auricular lymph nodes were found in the higher dose groups. No macroscopic abnormalities of the surrounding area were noted in any of the animals.

Interpretation of results:

Category 1 (skin sensitising) based on GHS criteria

Conclusions:

In LLNA study, FAT 40854/A is considered as skin sensitizer.

Executive summary:

In a GLP-compliant study, assessment of Contact Hypersensitivity to FAT 40854/A TE in the Mouse (Local Lymph Node Assay) was carried out according to OECD guideline 429 and EU method B42. Test substance concentrations selected for the main study were based on the results of a pre-screen test. In the main study, three experimental groups of five female CBA/J mice were treated with test substance concentrations of 10, 25 or 50 % w/w on three consecutive days, by open application on the ears. Five vehicle control animals were similarly treated, but with vehicle alone (Dimethyl formamide). Three days after the last exposure, all animals were injected with ³H-methyl thymidine and after five hours the draining (auricular) lymph nodes were excised and pooled for each animal. After precipitating the DNA of the lymph node cells, radioactivity measurements were performed. The activity was expressed as the number of Disintegrations Per Minute (DPM) and a stimulation index (SI) was subsequently calculated for each group. Red test substance remnants prevented scoring for erythema. No mortality occurred and no clinical signs of systemic toxicity were observed in the animals of the main study. Body weights and body weight gain of experimental animals remained in the same range as controls over the study period. All auricular lymph nodes of the animals of the experimental groups were considered larger in size compared to normal. The largest auricular lymph nodes were found in the higher dose groups No macroscopic abnormalities of the surrounding area were noted in any of the animals. Mean DPM/animal values for the experimental groups treated with test substance concentrations 10, 25 and 50 % were 5481, 6308 and 9451 DPM, respectively. The mean DPM/animal value for the vehicle control group was 470 DPM. The SI values calculated for the substance concentrations 10, 25 and 50 % were 11.7, 13.4 and 20.1, respectively. These results show that the test substance elicits an SI ≥3. The EC3 value (the estimated test substance concentration that will give a SI =3) was established to be lower than 10 %. Based on these results FAT 40854/A would be regarded as skin sensitizer.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Additional information:

In a GLP-compliant study, assessment of Contact Hypersensitivity to FAT 40854/A in the mouse (Local Lymph Node Assay) was carried out according to OECD guideline 429 and EU method B42. Test substance concentrations selected for the main study were based on the results of a pre-screen test. In the main study, three experimental groups of five female CBA/J mice were treated with test substance concentrations of 10, 25 or 50 % w/w on three consecutive days, by open application on the ears. Five vehicle control animals were similarly treated, but with vehicle alone (Dimethyl formamide). Three days after the last exposure, all animals were injected with ³H-methyl thymidine and after five hours the draining (auricular) lymph nodes were excised and pooled for each animal. After precipitating the DNA of the lymph node cells, radioactivity measurements were performed. The activity was expressed as the number of Disintegrations Per Minute (DPM) and a stimulation index (SI) was subsequently calculated for each group. Red test substance remnants prevented scoring for erythema. No mortality occurred and no clinical signs of systemic toxicity were observed in the animals of the main study. Body weights and body weight gain of experimental animals remained in the same range as controls over the study period. All auricular lymph nodes of the animals of the experimental groups were considered larger in size compared to normal. The largest auricular lymph nodes were found in the higher dose groups. No macroscopic abnormalities of the surrounding area were noted in any of the animals. Mean DPM/animal values for the experimental groups treated with test substance concentrations 10, 25 and 50 % were 5481, 6308 and 9451 DPM, respectively. The mean DPM/animal value for the vehicle control group was 470 DPM. The SI values calculated for the substance concentrations 10, 25 and 50 % were 11.7, 13.4, and 20.1, respectively. These results show that the test substance elicits an SI ≥3. The EC3 value (the estimated test substance concentration that will give a SI =3) was established to be lower than 10 %. Based on these results FAT 40854/A would be regarded as skin sensitizer.

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the findings in the skin sensitisation study, the test substance should be classified as Skin Sensitiser according to EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.