Formaldehyde, oligomeric reaction products with acetone and diphenylamine

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11 July 2016 to 05 August 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

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 the start of this in vivo skin irritation study in the rabbit. As recommended in the test guidelines, all available information was evaluated (e.g. existing human and animal data, literature, item data supplied by the Sponsor, analysis of structure activity relationships (SAR), physicochemical properties and reactivity (pH, buffering capacity) and in vitro (BCOP 508841; skin corrosion 508839; skin irritation 508839), ex-vivo and in vivo tests) to determine the need for in vivo skin testing. It was concluded that there is need to perform this in vivo skin irritation study in rabbit in order to establish the possible skin irritating properties of the test item.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

Organization for Economic Co-operation and Development (OECD), OECD Guidelines for Testing of Chemicals, Section 4, Health Effects, No.404: "Acute Dermal Irritation / Corrosion", Paris, 2015.

Deviations:

yes

Remarks:

see "Any other information"

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Commission Regulation (EC) No 440/2008 Part B: Methods for the Determination of Toxicity and other Health Effects; B4: "Acute Toxicity: Dermal Irritation/Corrosion". Official Journal of the European Union No. L142, May 2008, including most recent amendments.

Deviations:

yes

Remarks:

see "Any other information"

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Version / remarks:

United States Environmental Protection Agency (EPA). Health Effects Test Guidelines, OPPTS 870.2500, Acute Dermal Irritation. Office of Prevention, Pesticides and Toxic Items (7101), EPA 712-C-98-196, August 1998.

Deviations:

yes

Remarks:

see "Any other information"

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Formaldehyde, oligomeric reaction products with acetone and diphenylamine

EC Number:

500-011-5

EC Name:

Formaldehyde, oligomeric reaction products with acetone and diphenylamine

Cas Number:

9003-80-9

Molecular formula:

UVCB substance - not applicable

IUPAC Name:

N-phenylaniline; formaldehyde; propan-2-one

Test material form:

solid: flakes

Details on test material:

6.1. Test item
6.1.1. Test item information
Identification Formaldehyde, oligomeric reaction products with acetone
and diphenylamine
Appearance Dark brown flakes
Batch IC5B04P006
Purity/Composition 100% Unknown or Variable compositions, Complex reaction products and Biological materials (UVCB)
Test item storage At room temperature
Stable under storage conditions until 26 February 2019 (expiry date)

Specific details on test material used for the study:

Test substance: 206534/AIdentification: Formaldehyde, oligomeric reaction products with acetone and diphenylamineAppearance: Dark brown flakesBatch: IC5B04P006Purity/Composition: 100% Unknown or Variable compositions, Complex reaction products and Biological materials (UVCB)Test substance storage: At room temperatureStable under storage conditions until: 26 February 2019 (expiry date)Chemical names (IUPAC), synonym or trade name: Formaldehyde, oligomeric reaction products with acetone and diphenylamine (BXA)CAS Number: 9003-80-9pH (1% in water, indicative range): 7.76 – 7.37 (determined by Charles River Den Bsoch)

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Species: Albino rabbit, New Zealand White, (SPF-Quality). Recognized by international guidelines as the recommended test system (e.g. EC, OECD) Source: Charles River France, L’Arbresle Cedex, France Number of animals: 3 Males. Age and body weight: At start of dosing, the animals were between 12 and 24 weeks old and body weights were at least 1.5 kg. Identification: Earmark. Health inspection: At least prior to dosing. It was ensured that the animals were healthy and that the skin to be treated was intact and free from any abnormality.Conditions: Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, at least 10 air changes/hour, and a 12-hour light/12-hour dark cycle: the photoperiod was between 07:00 and 19:00 hrs daily. Any variations to these conditions were maintained in the raw data and had no effect on the outcome of the study. Accommodation: Animals were individually housed in labeled cages with perforated floors (Ebeco, Germany, dimensions 67 x 62 x 55 cm) and shelters (Ebeco, Germany, dimensions 40 x 32 x 23 cm). Acclimatization period was at least 5 days before start of treatment under laboratory conditions. Diet: Pelleted diet for rabbits (Global Diet 2030 from Harlan Teklad®, Mucedola, Milanese, Italy) approximately 100 grams per day. Hay (TecniLab-BMI BV, Someren, The Netherlands) and wooden sticks (Swedish aspen wood, Bioservices, Uden, The Netherlands) were available during the study period. Water: Free access to tap water. Diet, water, bedding and cage enrichment evaluation for contaminants and/or nutrients was performed according to facility standard procedures. There were no findings that could interfere with the study.

Test system

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

other: watery ethanol (50% v/v)

Controls:

not required

Amount / concentration applied:

Each animal was treated by dermal application of 0.5 grams of the test item.

Duration of treatment / exposure:

Four hours

Observation period:

72 hours

Number of animals:

3 animalsThe study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel). The two other animals were treated in a similar manner 20 days later, after considering the degree of skin irritation observed in the first animal.

Details on study design:

Test Item Preparation The test item was ground to a powder using a pestle and mortar prior to weighing and since the test item did not mix with water, the test item was moistened with watery ethanol (50% v/v) (water: Elix, Millipore S.A.S., Molsheim, France; ethanol: Merck, Darmstadt, Germany), immediately before application, to ensure close contact with the animal's skin. No corrections were made for the purity/composition of the test item, since the guidelines require a fixed amount that has to be applied.Treatment Approximately 24 hours before treatment, the dorsal fur was clipped with electric clippers, exposing an area of approximately 150 square centimetres (10x15 cm). To facilitate scoring, treated skin areas were re-clipped at least 3 hours before the observations. Each animal was treated by dermal application of 0.5 grams of the test item. The test item was moistened with 0.6 mL of the vehicle and applied to the skin of one flank, using a metalline patch of 2x3 cm. The patch was mounted on Micropore tape, which was wrapped around the abdomen and secured with Coban elastic bandage#. Four hours after the application, the dressing was removed and the skin cleaned of residual test item using tap water. After the final observation, the first animal was sacrificed by intra-venous injection of Euthasol® 20% (AST Farma BV, Oudewater, The Netherlands). The other two animals were removed from the study alive.Observations Mortality/Viability: Twice daily. Toxicity: At least once daily. Body Weight: Day of treatment (prior to application) and on the day of the final observation. Irritation: The skin reactions were assessed at approximately 1, 24, 48 and 72 hours after the removal of the dressings and test item. The irritation scores and a description of all other (local) effects were recorded. Adjacent areas of the untreated skin of each animal served as controls.Necropsy: No necropsy was performed according to study plan.Histopathology: No histopathology was performed.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No skin irritation was caused by 4 hours exposure to Formaldehyde, oligomeric reaction products with acetone and diphenylamine.There was no evidence of a corrosive effect on the skin.

Other effects:

Colouration / Remnants: Following exposure, light grey staining of the treated skin area was noted after removal of the test item. No further staining of the treated skin by the test item was observed and no test item remnants were seen. Clinical signs / Mortality: No signs of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Any other information on results incl. tables

Individual Skin Irritation Scores

Following exposure, light grey staining of the treated skin area was noted after removal of the test item

Animal	123^1			143			144
Time after exposure	Erythema (0-4)	Oedema (0-4)	Comments	Erythema (0-4)	Oedema (0-4)	Comments	Erythema (0-4)	Oedema (0-4)	Comments
1 hour 24 hours 48 hours 72 hours	0 0 0 0	0 0 0 0	- - - -	0 0 0 0	0 0 0 0	- - - -	0 0 0 0	0 0 0 0	- - - -

¹Sentinel

 

Mean Value Irritation Scores

Animal	Mean 24, 48 and 72 hrs
	Erythema	Oedema
123 143 144	0.0 0.0 0.0	0.0 0.0 0.0

 

Animal Specifications

Animal	Sex	Age at start   (weeks)	Body weights (grams)
			Prior to application	After the final observation
123 143 144	M M M	14 12 12	3192 2812 2686	3337 2883 2894

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

No skin irritation was caused by 4 hours exposure to Formaldehyde, oligomeric reaction products with acetone and diphenylamine.

Executive summary:

Primary skin irritation/corrosion study with Formaldehyde, oligomeric reaction products with acetone and diphenylamine in the rabbit (4-hour semi-occlusive application).

The study was carried out based on the guidelines described in:

OECD No.404, "Acute Dermal Irritation/Corrosion" (2015)

EC, No 440/2008; B4: "Acute Toxicity: Dermal Irritation/Corrosion".

US EPA, OPPTS 870.2500 (1998), Acute Dermal Irritation.

JMAFF Guidelines (2000), including the most recent revisions.

 

Three rabbits were exposed to 0.5 grams of Formaldehyde, oligomeric reaction products with acetone and diphenylamine, moistened with 50% watery ethanol by application onto clipped skin for 4 hours using a semi-occlusive dressing. Skin reactions were assessed 1, 24, 48 and 72 hours after exposure.

 

No skin irritation was caused by 4 hours exposure to Formaldehyde, oligomeric reaction products with acetone and diphenylamine.

 

Based on these results Formaldehyde, oligomeric reaction products with acetone and diphenylamine does not have to be classified and has no obligatory labelling requirement for skin irritation according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).