4,4'-Isopropylidenediphenol, polymer with 1-chloro-2,3-epoxypropane, propane-1,2-diol acrylate and succinic anhydride

Endpoint summary

Description of key information

Based on the results of an in vivo LLNA study, the test substance is considered to be sensitizing to skin

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:

From 06 June 2012 to 25 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

Type of study:

mouse local lymph node assay (LLNA)

Species:

mouse

Strain:

other: 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 (19 - 24 grams)
- Housing: Animals were group housed in labeld 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
Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, approximately 15 room air changes/hour, and a 12-hour light/12-hour dark cycle. Any variations to these conditions were maintained in the raw data and had no effect on the outcome of the study.

IN-LIFE DATES: From: 06 June to 25 June 2012

Concentration:

0, 10, 30 and 60%

No. of animals per dose:

5

Details on study design:

The vehicle was selected based on trial formulations performed at WIL Research Europe and on test substance data supplied by the sponsor.

RANGE FINDING TESTS:
In the interest of animal welfare and to minimize any testing likely to produce severe responses in animals, a weight of evidence analysis was performed prior to start of this study. All available information was evaluated (e.g. existing human and animal data, literature, substance data supplied by the sponsor, analysis of structure activity relationships (SAR), physicochemical properties and reactivity (pH, buffering capacity).

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: Local Lymph Node Assay
- Criteria used to consider a positive response:DPM values are presented for each animal and for each dose group. A Stimulation Index (SI) is calculated for each group. The SI is the ratio of the DPM/group compared to DPM/vehicle control group. If the results indicate a SI ≥ 3, the test substance may be regarded as a skin sensitizer. The results were evaluated according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (20011) and the Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008 on Classification, Labelling and Packaging of substances and mixtures.

ANIMAL ASSIGNMENT
Three groups of five animals were treated with one test substance concentration per group. One group of five animals was treated with vehicle.

TREATMENT PREPARATION AND ADMINISTRATION:
Test substance preparation: The test substance formulations (w/w) were prepared within 4 hours prior to each treatment. Homogeneity was obtained to visually acceptable levels.
Rationale for vehicle: The vehicle was selected based on trial formulations performed at NOTOX and on test substance data supplied by the sponsor.

Induction - Days 1, 2 and 3; Excision of nodes - Day 6; Tissue processing for radioacitivity - Day 6; Radioactivity measurements - Day 7; Performed according to test guidelines.

Observations:
Mortality/Viability: Twice daily.
Body weights: On Day 1 (pre-dose) and Day 6 (prior to necropsy).
Clinical signs: Once daily on Days 1-6 (on Days 1-3 between 3 and 4 hours after dosing).
Irritation: Once daily on Days 1-6 (on Days 1 - 3 immediately after dosing) according to the following numerical scoring system. Furthermore, a description of all other (local) effects was recorded according to guidelines.

Necropsy: All animals surviving to the end of the study were sacrificed by intra-peritoneal injection with Euthasol® 20% (0.2 mL/animal).

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'.

Key result

Parameter:

SI

Value:

1

Test group / Remarks:

at 10% test concentration

Key result

Parameter:

SI

Value:

1.5

Test group / Remarks:

at 30% test concentration

Key result

Parameter:

SI

Value:

5.1

Test group / Remarks:

at 60% test concentration

Key result

Parameter:

EC3

Value:

42.5

Test group / Remarks:

test concentration estimated to produce SI =3

Results Pre-screen test:

Well-defined erythema was noted in both animals at 60% on Days 2-5 (very slight on Day 6) and very slight erythema was noted in both animals at 30% on Days 2-6. Variations in ear thickness during the observation period were less than 30% from Day 1 pre-dose values. All animals showed bald areas on the head from Day 3 onwards.

Based on these results, the highest test substance concentration selected for the main study was a 60% concentration.

Other results - main study:

Skin reactions / Irritation:

The very slight irritation, or up to well-defined irritation in case of the 60% treated animals, of the ears as shown by all animals was considered not to have a toxicologically significant effect on the activity of the nodes.All animals at 60% showed bald areas around the ears on Day 5 and 6. Based on the pre-screen data and the maximal erythema grade 2, this bald skin is considerednot to have a toxicologically significant effect on the activity of the nodes.

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. The body weight loss noted for some animals across the dose groups was considered not toxicologically significant since the changes were slight in nature and no concentration-related incidence was apparent.

Macroscopy of the auricular lymph nodes and surrounding area:

All auricular lymph nodes of the animals treated at 60% were considered larger in size, as compared to the other treated groups. No macroscopic abnormalities of the surrounding area were noted in any of the animals.

Interpretation of results:

other: sensitising

Conclusions:

Under the conditions of the study, the test substance was therefore considered to be sensitizing to skin

Executive summary:

A study was conducted to determine the skin sensitization potential of the test substance '4,4'-Isopropylidenediphenol, polymer with 1-chloro-2,3-epoxypropane, propane-1,2-diol acrylate and succinic anhydride'in mouse using the local lymph node assay according to OECD Guideline 429 and EU Method B.42, in compliance with GLP. The 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, 30 or 60% w/w on three consecutive days, by open application on the ears. Five vehicle control animals were similarly treated, but with vehicle alone (Methyl ethyl ketone). 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.The very slight irritation, or up to well-defined irritation in case of the 60% treated animals, of the ears as shown by all animals was considered not to have a toxicologically significant effect on the activity of the nodes. All animals at 60% showed bald areas around the ears on Day 5 and 6. Based on the pre-screen data and the maximal erythema grade 2, this bald skin is considered not to have a toxicologically significant effect on the activity of the nodes.No mortality occurred and no clinical signs of systemic toxicity were observed. Body weights and body weight gain of experimental animals remained in the same range as controls over the study period. The body weight loss noted for some animals across the dose groups was considered not toxicologically significant since the changes were slight in nature and no concentration-related incidence was apparent.All auricular lymph nodes of the animals treated at 60% were considered larger in size, as compared to the other treated groups. No macroscopic abnormalities of the surrounding area were noted in any of the animals. The SI values calculated for the substance concentrations 10, 30 and 60% were 1.0, 1.5 and 5.1 respectively. These results indicate that the test substance could elicit an SI ≥ 3. An EC3 value (the estimated test substance concentration that will give a SI =3) of 42.5% was calculated. Under the conditions of the study, the test substance was therefore considered to be sensitizing to skin (WIL Research Europe, 2012).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Additional information:

A study was conducted to determine the skin sensitization potential of the test substance '4,4'-Isopropylidenediphenol, polymer with 1-chloro-2,3-epoxypropane, propane-1,2-diol acrylate and succinic anhydride' in mouse using the local lymph node assay according to OECD Guideline 429 and EU Method B.42, in compliance with GLP. The 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, 30 or 60% w/w on three consecutive days, by open application on the ears. Five vehicle control animals were similarly treated, but with vehicle alone (Methyl ethyl ketone). 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.The very slight irritation, or up to well-defined irritation in case of the 60% treated animals, of the ears as shown by all animals was considered not to have a toxicologically significant effect on the activity of the nodes. All animals at 60% showed bald areas around the ears on Day 5 and 6. Based on the pre-screen data and the maximal erythema grade 2, this bald skin is considered not to have a toxicologically significant effect on the activity of the nodes.No mortality occurred and no clinical signs of systemic toxicity were observed. Body weights and body weight gain of experimental animals remained in the same range as controls over the study period. The body weight loss noted for some animals across the dose groups was considered not toxicologically significant since the changes were slight in nature and no concentration-related incidence was apparent.All auricular lymph nodes of the animals treated at 60% were considered larger in size, as compared to the other treated groups. No macroscopic abnormalities of the surrounding area were noted in any of the animals. The SI values calculated for the substance concentrations 10, 30 and 60% were 1.0, 1.5 and 5.1 respectively. These results indicate that the test substance could elicit an SI ≥ 3. An EC3 value (the estimated test substance concentration that will give a SI =3) of 42.5% was calculated. Under the conditions of the study, the test substance was therefore considered to be sensitizing to skin (WIL Research Europe, 2012).

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the results of an in vivo LLNA study as well as its calculated EC3 value, the test substance qualifies for classification as Skin sens. Cat 1B (H317 – May cause an allergic skin reaction) according to CLP (EC 1272/2008) criteria.