Formaldehyde, oligomeric reaction products with acetone and diphenylamine

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25 July 2016 to 09 August 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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 402 (Acute Dermal Toxicity)

Version / remarks:

Organization for Economic Co-operation and Development (OECD), OECD Guidelines for Testing of Chemicals, Section 4, Health Effects, No. 402, "Acute Dermal Toxicity", Paris, 1987.

Deviations:

yes

Remarks:

See "Any other information"

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Version / remarks:

Commission Regulation (EC) No 440/2008 Part B: Methods for the Determination of Toxicity and other Health Effects; B.3: "Acute Toxicity (Dermal)". 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 870.1200 (Acute Dermal Toxicity)

Version / remarks:

United States Environmental Protection Agency (EPA). Health Effects Test Guidelines, OPPTS 870.1200, Acute Dermal Toxicity. Office of Prevention, Pesticides and Toxic Items (7101), EPA 712-C-98-192, August 1998.

Deviations:

yes

Remarks:

see "Any other information"

GLP compliance:

yes

Test type:

standard acute method

Limit test:

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 Viable compositions, Complex reaction products and Biological materials (UVCB)Test substance storage: At room temperatureStable under storage conditions until: 26 February 2019 (expiry date)Chemical name (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 Bosch)

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Species: Rat, Wistar strain, Crl:WI (Han) (outbred, SPF-Quality). Recognized by international guidelines as the recommended test system (e.g. OECD, EC). Source: Charles River Deutschland, Sulzfeld, Germany. Number of animals: 5 males and 5 females (females were nulliparous and non-pregnant). Age and body weight: Young adult animals (approx. 10 weeks old) were selected. Body weight variation did not exceed +/- 20% of the sex mean. Identification: Tail mark with indelible ink. 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: Individually housed in labeled Makrolon cages (MIII type, height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom). Acclimatization period was at least 5 days before start of treatment under laboratory conditions. During the acclimatization period the animals were group housed in Makrolon cages (MIV type, height 18 cm). Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany). 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.

Administration / exposure

Type of coverage:

occlusive

Vehicle:

propylene glycol

Details on dermal exposure:

Test Item Preparation Vehicle: Propylene glycol (Merck, Darmstadt, Germany) (specific gravity 1.036) Rationale: The vehicle was selected based on trial preparation performed at Charles River Den Bosch and on test item data supplied by the Sponsor. There was no information available regarding the solubility or stability in vehicle. Preparation: The preparation (w/w) was kept at room temperature and dosed within 4 hours after adding the vehicle to the test item. Homogeneity was assessed by visual inspection of the solutions and the formulations were stirred during dosing, which ensures homogeneity sufficient for these kinds of studies. Adjustment was made for specific gravity of the vehicle. No correction was made for purity of the test item.Treatment Method: Dermal application. The test item (preparation) was stirred on a magnetic stirrer during application. Clipping One day before exposure (Day -1) an area of approximately 5x7 cm on the back of each animal was clipped. Application: The test item preparation was applied on an area of approx. 10% of the total body surface, i.e. approx. 25 cm² for males and 18 cm² for females. The test item preparation was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D), successively covered with aluminum foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only.Application: period 24 hours, after which dressings were removed and the skin cleaned of residual test item using tap water. The skin was dried by gentle padding using a tissue.

Duration of exposure:

24 hours

Doses:

2000 mg/kg (10 mL/kg) body weight.

No. of animals per sex per dose:

10 animals (5 male/5 female)

Control animals:

not required

Details on study design:

Mortality/Viability: Twice daily. Body weights: Days 1 (pre-administration), 8 and 15. Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15. The time of onset, degree and duration were recorded and the symptoms graded according to fixed scales: Maximum grade 4: grading slight (1) to very severe (4) Maximum grade 3: grading slight (1) to severe (3) Maximum grade 1: presence is scored (1). Necropsy: At the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.

Statistics:

Not specified

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

Flat posture, ptosis and/or chromodacryorrhoea (snout) were noted for all animals on Day 1.Black discoloration was seen in the treated skin-area of all animals on Days 2 and/or 3. This was considered due to the colour of the formulation. Scales and/or scabs were noted for animals no. 3, 4, 5 and 9 between Days 5 and 15. These local effects were considered not to have affected the conclusion of the study.

Body weight:

The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Other findings:

No further findings specified in the study report

Any other information on results incl. tables

MORTALITY DATA

TEST DAY	1	1	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15
HOURS AFTER TREATMENT	0	2	4
MALES 2000 MG/KG	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
FEMALES 2000 MG/KG	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-

 

CLINICAL SIGNS

TEST DAY		1	1	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15
HOURS AFTER TREATMENT	MAX GRADE	0	2	4
MALES 2000 MG/KG
ANIMAL 1 Posture                Flat posture Various                Ptosis                Black (Treated skin)	(1)   (3) (1)	-   - -	-   - -	1   3 -	-   - 1	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -
ANIMAL 2 Posture                Flat posture Various                Ptosis                Black (Treated skin)	(1)   (3) (1)	-   - -	-   - -	1   2 -	-   - 1	-   - 1	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -
ANIMAL 3 Posture                Flat posture Skin / fur                Scales (Treated skin) Various                Ptosis                Black (Treated skin)	(1)   (3)   (3) (1)	-   -   - -	-   -   - -	1   -   2 -	-   -   - 1	-   -   - -	-   -   - -	-   1   - -	-   1   - -	-   1   - -	-   1   - -	-   1   - -	-   1   - -	-   1   - -	-   -   - -	-   -   - -	-   -   - -	-   -   - -
ANIMAL 4 Posture                Flat posture Skin / fur                Scales (Treated skin)                Scabs Secretion / excretion                Chromodacryorrhoea (Snout) Various                Ptosis                Black (Treated skin)	(1)   (3) (3)   (3)   (3) (1)	-   - -   1   - -	-   - -   1   - -	1   - -   1   3 -	-   - -   -   - 1	-   - -   -   - 1	-   - -   -   - -	-   1 -   -   - -	-   1 -   -   - -	-   1 -   -   - -	-   1 -   -   - -	-   1 1   -   - -	-   1 1   -   - -	-   1 1   -   - -	-   - 1   -   - -	-   - 1   -   - -	-   - 1   -   - -	-   - 1   -   - -
ANIMAL 5 Posture                Flat posture Skin / Fur                Scales (Treated skin) Various                Ptosis                Black (Treated skin)	(1)   (3)   (3) (1)	-   -   - -	-   -   - -	1   -   3 -	-   -   - 1	-   -   - 1	-   -   - -	-   -   - -	-   1   - -	-   1   - -	-   1   - -	-   1   - -	-   1   - -	-   1   - -	-   1   - -	-   -   - -	-   -   - -	-   -   - -
FEMALES 2000 MG/KG
ANIMAL 6 Posture                Flat posture Secretion / excretion                Chromodacryorrhoea (Snout) Various                Ptosis                Black (Treated skin)	(1)   (3)   (3) (1)	-   -   - -	-   1   - -	1   1   3 -	-   -   - 1	-   -   - -	-   -   - -	-   -   - -	-   -   - -	-   -   - -	-   -   - -	-   -   - -	-   -   - -	-   -   - -	-   -   - -	-   -   - -	-   -   - -	-   -   - -
ANIMAL 7 Posture                Flat posture Various                Ptosis                Black (Treated skin)	(1)   (3) (1)	-   - -	-   - -	1   3 -	-   - 1	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -
ANIMAL 8 Posture                Flat posture Various                Ptosis                Black (Treated skin)	(1)   (3) (1)	-   - -	-   - -	1   2 -	-   - 1	-   - 1	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -
ANIMAL 9 Posture                Flat posture Skin / fur                Scales (Treated skin)                Scabs Various                Ptosis                Black (Treated skin)	(1)   (3) (3)   (3) (1)	-   - -   - -	-   - -   - -	1   - -   2 -	-   - -   - 1	-   - -   - 1	-   - -   - -	-   1 -   - -	-   1 -   - -	-   1 -   - -	-   1 -   - -	-   1 1   - -	-   1 1   - -	-   1 1   - -	-   1 1   - -	-   1 1   - -	-   1 1   - -	-   1 1   - -
ANIMAL 10 Posture                Flat posture Various                Ptosis                Black (Treated skin)	(1)   (3) (1)	-   - -	-   - -	1   3 -	-   - 1	-   - 1	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -	-   - -

- = sign not observed

 

BODY WEIGHTS (GRAM)

SEX/DOSE LEVEL	ANIMAL	DAY 1	DAY 8	DAY 15
MALES 2000 MG/KG
	1 2 3 4 5	276 286 276 277 284	286 291 287 278 300	303 314 306 291 327
	MEAN ST. DEV. N	280 5 5	288 8 5	308 13 5
FEMALES 2000 MG/KG
	6 7 8 9 10	182 182 189 189 186	181 198 186 194 188	195 206 192 210 191
	MEAN ST. DEV. N	186 4 5	189 7 5	199 9 5

 

MACROSCOPIC FINDINGS

ANIMAL	ORGAN	FINDING	DAY OF DEATH
MALES 2000 MG/KG
1		No findings noted	Scheduled necropsy Day 15 after treatment
2		No findings noted	Scheduled necropsy Day 15 after treatment
3		No findings noted	Scheduled necropsy Day 15 after treatment
4		No findings noted	Scheduled necropsy Day 15 after treatment
5		No findings noted	Scheduled necropsy Day 15 after treatment
FEMALES 2000 MG/KG
6		No findings noted	Scheduled necropsy Day 15 after treatment
7		No findings noted	Scheduled necropsy Day 15 after treatment
8		No findings noted	Scheduled necropsy Day 15 after treatment
9		No findings noted	Scheduled necropsy Day 15 after treatment
10		No findings noted	Scheduled necropsy Day 15 after treatment

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The dermal LD50 value of Formaldehyde, oligomeric reaction products with acetone and diphenylamine in Wistar rats was established to exceed 2000 mg/kg body weight.

Executive summary:

Assessment of acute dermal toxicity with Formaldehyde, oligomeric reaction products with acetone and diphenylamine in the rat.

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

OECD No.402 (1987) "Acute Dermal Toxicity"

Commission Regulation (EC) No 440/2008, B3: "Acute Toxicity (Dermal)"

EPA, OPPTS 870.1200 (1998), "Acute Dermal Toxicity"

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

 

Formaldehyde, oligomeric reaction products with acetone and diphenylamine was administered to five Wistar rats of each sex by a single dermal application at 2000 mg/kg body weight for 24 hours. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15).

 

No mortality occurred.

Flat posture, ptosis and/or chromodacryorrhoea (snout) were noted for all animals on Day 1.

Black discoloration was seen in the treated skin-area of all animals on Days 2 and/or 3. This was considered due to the colour of the formulation. Scales and/or scabs were noted for animals no. 3, 4, 5 and 9 between Days 5 and 15. These local effects were considered not to have affected the conclusion of the study.

The mean body weight gain during the observation period was within the range expected for rats used in this type of study.

No abnormalities were found at macroscopic post mortem examination of the animals.

 

The dermal LD50 value of Formaldehyde, oligomeric reaction products with acetone and diphenylamine in Wistar rats was established to exceed 2000 mg/kg body weight.

 

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 acute dermal toxicity 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).