4,4'-benzylidenedianiline

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25 November 2009 - 04 December 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

24 April 2002

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Version / remarks:

No. 440/2008

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

4,4'-benzylidenedianiline

EC Number:

210-038-1

EC Name:

4,4'-benzylidenedianiline

Cas Number:

603-40-7

Molecular formula:

C19H18N2

IUPAC Name:

4-[(4-aminophenyl)(phenyl)methyl]aniline

Test material form:

solid

Details on test material:

Identification: DATIPH
Physical form: Solid
Solubility in water: < 1 g/L
Colour: yellow brown
CAS no.: 603-40-7
Chemical name: 4,4’-Benzylidenedianiline
Molecular formula: C19H18N2
Test substance storage: At room temperature protected from light
Stability: Stable under storage conditions
Expiry date: 31 October 2010

Specific details on test material used for the study:

- Source of test material: Sponsor (identification DATIPH)
- Description: brown solid lumps 09 November 2009
- Date received: 09 November 2009
- Storage conditions: approximately 4°C in the dark

The integrity of supplied data relating to the identity, purity and stability of the test material is the responsibility of the Sponsor.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: For the purpose of the study the test material was ground to a powder prior to application.
- Preliminary purification step (if any):
- Final dilution of a dissolved solid, stock liquid or gel:
- Final preparation of a solid:

FORM AS APPLIED IN THE TEST (if different from that of starting material)

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable)

OTHER SPECIFICS:
The absorption of the test material was not determined.
The pH of the test material was determined prior to commencement of the study and found to be as follows:

Preparation pH Measurement
immediately after 10 minutes after 20 minutes
10% w/w aqueous preparation of the test material 8.9 9.4 9.5

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories UK Limited, Hillcrest, Belton, Loughborough, UK
- Age at study initiation: twelve to twenty weeks old
- Weight at study initiation: 2.38 or 2.39 kg
- Housing: individually housing in suspended cages
- Diet (e.g. ad libitum): 2030 Teklad Global Rabbit diet supplied by Harlan Teklad, Blackthorn, Bicester, Oxon, UK
- Water (e.g. ad libitum): 2030 Teklad Global Rabbit diet supplied by Harlan Teklad, Blackthorn, Bicester, Oxon, UK
The diet and drinking water were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23°C
- Humidity (%): 30 to 70%
- Air changes (per hr): at least fifteen changes per hour
- Photoperiod (hrs dark / hrs light): twelve hours continuous light (06:00 to 18:00) and twelve hours darkness

The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Remarks:

On the day before the test each rabbit was clipped free of fur from the dorsal/flank area using veterinary clippers.

Vehicle:

water

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g of the test material, moistened sufficiently with 0.5 ml of distilled water to achieve a paste

Duration of treatment / exposure:

One animal with three patches for a duration of 3 minutes, 1 hour and 4 hours, respectively.
The second animal with one patch for 4 hours.

Observation period:

lmmediately following removal of the patches and approximately 1, 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored according to the following scale. The proposed 24 and 48-hour observations at the 3 minute and 1- hour exposure sites were performed approximately three to four hours later than specified in the guideline.

Number of animals:

2

Details on study design:

TEST SITE
- Area of exposure: three suitable sites on the back of the rabbit
- % coverage: area under the 2.5 cm x 2.5 cm cotton gauze patch
- Type of wrap if used: the trunk of the rabbit was wrapped in an elasticated corset for the duration of the exposure period

REMOVAL OF TEST SUBSTANCE
- Washing (if done): any residual test material was removed by gentle swabbing with cotton wool soaked in distilled water
- Time after start of exposure: one patch at each time point (3 minutes, 1 hour, 4 hours and after application)

OBSERVATION TIME POINTS
lmmediately following removal of the patches and approximately 1, 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored.

SCORING SYSTEM:
- Erythema and eschar formation and oedema formation were evaluated in the scale of severity from 0 to 4 (Table1).
- Method of calculation:
The scores for erythema and oedema at the 24 and 72-hour readings were totalled for the two test rabbits (8 values) and this total was divided by four to give the primary irritation index of the test material.
The test material was classified according to the scheme (Table 2) devised by Draize J H (1959) "Dermal Toxicity" ln: Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics. Association of Food and Drug Officiais of the United States, Austin, Texas, p.46-59:

If irreversible alteration of the dermal tissue is noted in any rabbit, as judged by the Study Director, which include ulceration and clear necrosis or signs of scar tissue, the test material is classified as corrosive to rabbit skin. Classification according to Draize may, therefore, not be applicable.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Remarks:

3-minute and 1-hour exposure

Basis:

animal #1

Time point:

other: Immediate, 1h, 28h, 52h, 72h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

edema score

Remarks:

3-minute and 1-hour exposure

Basis:

animal #1

Time point:

other: Immediate, 1h, 28h, 52h, 72h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

erythema score

Remarks:

4-hour exposure

Basis:

animal #1

Time point:

other: Immediate, 1h, 24h, 48h, 72h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

edema score

Remarks:

4-hour exposure

Basis:

animal #1

Time point:

other: Immediate, 1h, 24h, 48h, 72h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 5

Irritation parameter:

erythema score

Remarks:

4-hour exposure

Basis:

animal #2

Time point:

other: Immediate, 1h, 24h, 48h, 72h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 6

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: Immediate, 24h, 48h, 72h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

3-minute and 1-hour semi-occluded applications of the test material to the intact skin of one rabbit produced no evidence of skin irritation.
A single 4-hour semi-occluded application of the test material to the intact skin of two rabbits produced no evidence of skin irritation.

Other effects:

All animals showed expected gain in bodyweight during the study (Table 2).

Any other information on results incl. tables

Table 2. lndividual Bodyweights and Bodyweight Changes

Rabbit Number and Sex	lndividual Bodyweight (kg)		Bodyweight Change (kg)
	Day0	Day3
68684 Male	2.39	2.43	0.04
68696 Male	2.38	2.47	0.09

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test material produced a primary irritation index of 0.0 and was classified as non-irritant to rabbit skin according to the Draize classification scheme. The test material did not meet the criteria for classification as irritant or corrosive according to EU labelling regulations (EC) No 440/2008.

Executive summary:

The study was performed to assess the irritancy potential of the 4,4'-Benzylidenedianiline following single, 3-minute, 1 and 4-hour, semi-occluded applications to the intact rabbit skin according to the OECD guideline 404 and method B4 of the Commission Regulation (EC) No. 440/2008. None of the applications to the rabbit intact skin produced any evidence of skin irritation. The test material produced a primary irritation index of 0.0 and was classified as non-irritant to rabbit skin according to the Draize classification scheme. The test material did not meet the criteria for classification as irritant or corrosive according to EU labelling regulations (EC) No 440/2008.