4,4'-[2,2,2-trifluoro-1-(trifluoromethyl)ethylidene]diphenol; bisphenol AF

Endpoint summary

Description of key information

Non-sensitising; OECD 406 ; Drs. A.H.B.M. van Huygevoort., 2001c

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Reference 1

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19 February - 25 May 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study was conducted in accordance with International guidelines and in accordance with GLP. All guideline validity criteria were met.

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

The study pre-dates the adoption of the OECD 429 OECD Guideline (2002). The most appropriate method at the time was conducted for this study.

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature, dark.
- Stability under test conditions: Not reported
- Solubility and stability of the test substance in the solvent/vehicle: Not reported
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: Not reported

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Test item formulated in corn oil within fours prior to treatment.
- Preliminary purification step (if any): N/A
- Final dilution of a dissolved solid, stock liquid or gel: N/A
- Final preparation of a solid: N/A

FORM AS APPLIED IN THE TEST (if different from that of starting material): Liquid (formulated in corn oil)

OTHER SPECIFICS: Homogeneity of the formulation was confirmed by visual inspection.

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Kisslegg, Germany
- Females (if applicable) nulliparous and non-pregnant: Yes
- Microbiological status of animals, when known: Not reported
- Age at study initiation: ca. 7 weeks
- Weight at study initiation: 378 - 467 g
- Housing: Grouped housing of 5 animals per metal cage with wire-mesh floors.
- Diet (e.g. ad libitum): Free access to standard guinea pig diet, including ascorbic acid (1000 mg/kg) (Charles River Breeding and Maintenance Diet for Guinea Pigs, Altorim, Germany)
- Water (e.g. ad libitum): Free access to tap water
- Acclimation period: Minimum of 5 days
- Indication of any skin lesions: Not reported

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 3 ºC
- Humidity (%): 30 - 70 %
- Air changes (per hr): ca. 15
- Photoperiod (hrs dark / hrs light): 12:12 light:dark
- IN-LIFE DATES: From: To: Not reported

Route:

intradermal

Vehicle:

corn oil

Concentration / amount:

1 %

Day(s)/duration:

Day 1

Adequacy of induction:

highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically

Route:

epicutaneous, occlusive

Vehicle:

corn oil

Concentration / amount:

50% / 0.5 mL

Day(s)/duration:

Day 8 / 48 hrs

Adequacy of induction:

highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

corn oil

Concentration / amount:

5 % / 0.1 mL

Day(s)/duration:

Day 21 / 24 hrs

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

Treated: 10
Control: 5

Details on study design:

RANGE FINDING TESTS:

Formulations of the test item were prepared in corn oil.

Intradermal test- A series of 6 test item concentrations were used; 20, 10, 5, 2, 1 and 0.5 % (top concentration being the highest that could technically be injected) with each of the three animals receiving two injections (0.1 mL) each i.e. 20 and 10 % tested in one animal, 5 and 2 % in the second etc. 1 % was found to cause moderate irritation and was chosen as the concentration to be used for the induction phase of the main test.

Epidermal test- Four concentrations were used; 50, 20, 10 and 5 % (top concentration being the highest that could be technically applied) with each of the four animals treated with two concentrations each i.e. test concentrations were tested in duplicate. 0.5 mL solution was applied to clipped flanks and covered for 24 h. Residual test item was then removed, and the skin cleaned using water. Observations were taken after 24 and 48 h contact time. The 5 % solution would be used in the challenge phase of the main test as it was the highest concentration that did not cause irritation. The 50 % solution would be used in the main test as it was the maximum concentration found to cause moderate irritation.

MAIN STUDY
A. INDUCTION EXPOSURE - Intradermal injection
- No. of exposures: 1
- Exposure period: Day 1
- Test groups: 10 animals each injected at 3 sites (injections in pairs either side of the midline) (1. mixture of Freunds' Complete Adjuvant (FCA) and water, 2. test item at 1 % concentration, 3. test item and FCA 1:1 v/v (1 % concentration of test item)
- Control group: 5 animals
- Site: Scapular region
- Frequency of applications: Once
- Duration: n/a
- Concentrations: 1 %

A. INDUCTION EXPOSURE - Epidermal injection
- No. of exposures: 1
- Exposure period: Day 8
- Test groups: 10 animals treated using a Metalling patch (2 x 3 cm) mounted on Medical tape, which was held in place with Micropore tape and subsequently Coban elastic bandage.
- Control group: 5 animals treated similarly except that vehicle alone was administered.
- Site: scapular region, between injection sites
- Frequency of applications: Once
- Duration: 48 hrs
- Concentrations: 1 %

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: Day 21
- Exposure period: 24 h
- Test groups: 10 animals treated using Patch Test Plasters held in place with Micropore tape and subsequently Coban elastic bandage.
- Control group: 5 animals treated similarly than controls.
- Site: flank
- Concentrations: 5 %
- Evaluation (hr after challenge): 24 and 48 h after application

Challenge controls:

Corn oil

Positive control substance(s):

yes

Remarks:

Assessment study conducted within last 6 months successfully validated procedure. Reference item was alpha-hexylcinnamicaldehyde. Intradermal induction: 5% solution in water. Epidermal induction: undiluted. Challenge: 10% solution in water.

Positive control results:

100 % sensitisation observed in test group

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

5 %

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

5 %

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

Control

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

Control

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Dose level:

10 % (alpha-hexylcinnamicaldehyde)

No. with + reactions:

10

Total no. in group:

10

Clinical observations:

None

Remarks on result:

positive indication of skin sensitisation

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Dose level:

10 % (alpha-hexylcinnamicaldehyde)

No. with + reactions:

10

Total no. in group:

10

Clinical observations:

None

Remarks on result:

positive indication of skin sensitisation

Table 1:       Erythema gradings after induction and challenge phases

Rep.	Intradermal			Epidermal
	3 days after induction			10 days after induction		23 days after induction (1 day after challenge)		24 days after induction (2 days after challenge)
	A (Erythema Score)	B (Erythema Score)	C (Erythema Score)	Erythema Score	Oedema Score	5 %	Control	5 %	Control
Control
1	3	2	2	2	0	0	0	0	0
2	3	2	3	0	0	0	0	0	0
3	3	3	3	0	0	0	0	0	0
4	3	2	3	2	0	0	0	0	0
5	3	1	3	2	0	0	0	0	0
Treatment
1	2	1	2	3	0	0	0	0	0
2	3	3	3	3	0	0	0	0	0
3	3	3	3	3	0	0	0	0	0
4	3	2	3	3	0	0	0	0	0
5	3	2	3	3	0	0	0	0	0
6	2	2	2	n	1	0	0	0	0
7	3	2	3	3	0	0	0	0	0
8	3	2	3	3	0	0	0	0	0
9	3	2	2	3	0	0	0	0	0
10	3	3	3	n	1	0	0	0	0

A: 1:1 mixture of FCA and water for injection

B: Corn oil (control) or 1 % test item (treatment)

C: Corn oil (control) or 1:1 mixture of FCA and test tem (1 % final concentration) (treatment)

Epidermal induction: Corn oil (control) or 50 % test item (treatment)

Epidermal challenge: Corn oil (control) or 5 % test item (treatment)

n: necrosis observed

Grading’s on a scale of 0 – 4 (0: no erythema / no oedema, 1: slight erythema / slight oedema, 2: well-defined erythema, 3: moderate erythema and swelling, 4: severe erythema, swelling and deep epidermal injury)

Interpretation of results:

GHS criteria not met

Conclusions:

The test item cannot be classified as a skin sensitiser based on GHS criteria.

Executive summary:

In a dermal sensitization study (OECD 406) with test item in corn oil, young adult Dunkin Hartley albino guinea pigs (15 females) were tested using the guinea pig maximisation method.

 

A preliminary irritation study was conducted using 3 individuals in order to select the test substance concentrations to be used in the main study. The results concluded that intradermal injections at a concentration of 1 % test item and epidermal application at a concentration of 50 % test item were the highest concentrations that produced moderate irritation. These concentrations were therefore assigned in the induction phase of the main test. The highest epidermal concentration applied that did not cause irritation was 5 % test item. This concentration was therefore assigned in the challenge phase of the main test.

 

The main test was conducted using 15 individuals, 5 controls and 10 treated. On Day 1, each individual was subjected to three pairs of intradermal injection around the scapular region (0.1 mL/ site); 2 x 1:1 FCA:water, 2 x 1 % test item in corn oil (or corn oil only for controls), 2 x 1 % test item in FCA (1:1 mix) (corn oil: FCA mix for controls). The dermal reactions caused by the injections were assessed on Day 3 for irritation. On Day 8, the scapular region was clipped and a dermal application (0.5 mL) of the test item (50 % concentration) was applied (2 x 3 cm patch) between the injection sites. The patch was held in place for 48 using micropore tape and an elastic bandage. The patches were removed, and the applications sites cleaned prior to assessment. On Day 21 each individual received a challenge dermal application, that was applied as previously described, at a concentration of 5 %. The patch was removed after 24 h contact time, the skin cleaned and assessment for irritation made at 24 and 48 h (after challenge application).

 

The results of the main studies showed moderate erythema (grade 3) at the injection and dermal patch sites after the induction of the test item at a concentration of 1 % and 50 %, respectively. It should be noted that signs of necrosis were observed in 2 treated individuals after the dermal induction phase. No skin reactions were observed following the challenge phase in any of the individuals. No mortality or systemic toxicity were observed during the main test and body weight increases in the treated individuals was in the same range as the controls.

 

In this study, the test item is not a dermal sensitizer.

Reference 2

Endpoint:

skin sensitisation: in vitro

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro skin sensitisation study does not need to be conducted because adequate data from an in vivo skin sensitisation study are available

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
An in vitro or in chemico skin sensitisation study does not need to be conducted because adequate data from an in vivo skin sensitisation study are available (carried out before 11 October 2016).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

A preliminary irritation study was conducted using 3 individuals in order to select the test substance concentrations to be used in the main study. The results concluded that intradermal injections at a concentration of 1 % test item and epidermal application at a concentration of 50 % test item were the highest concentrations that produced moderate irritation. These concentrations were therefore assigned in the induction phase of the main test. The highest epidermal concentration applied that did not cause irritation was 5 % test item. This concentration was therefore assigned in the challenge phase of the main test.

 

The main test was conducted using 15 individuals, 5 controls and 10 treated. On Day 1, each individual was subjected to three pairs of intradermal injection around the scapular region (0.1 mL/ site); 2 x 1:1 FCA:water, 2 x 1 % test item in corn oil (or corn oil only for controls), 2 x 1 % test item in FCA (1:1 mix) (corn oil: FCA mix for controls). The dermal reactions caused by the injections were assessed on Day 3 for irritation. On Day 8, the scapular region was clipped and a dermal application (0.5 mL) of the test item (50 % concentration) was applied (2 x 3 cm patch) between the injection sites. The patch was held in place for 48 using micropore tape and an elastic bandage. The patches were removed, and the applications sites cleaned prior to assessment. On Day 21 each individual received a challenge dermal application, that was applied as previously described, at a concentration of 5 %. The patch was removed after 24 h contact time, the skin cleaned and assessment for irritation made at 24 and 48 h (after challenge application).

 

The results of the main studies showed moderate erythema (grade 3) at the injection and dermal patch sites after the induction of the test item at a concentration of 1 % and 50 %, respectively. It should be noted that signs of necrosis were observed in 2 treated individuals after the dermal induction phase. No skin reactions were observed following the challenge phase in any of the individuals. No mortality or systemic toxicity were observed during the main test and body weight increases in the treated individuals was in the same range as the controls.

 

In this study, Bisphenol AF is not a dermal sensitiser.

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

The substance does not meet the classification criteria for skin sensitisation in accordance with Regulation (EC) No 1272/2008 (CLP).