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REACH

Formaldehyde, oligomeric reaction products with acetone and diphenylamine

EC number: 500-011-5 | CAS number: 9003-80-9

Life Cycle description
Uses advised against

Toxicological information

Endpoint summary

Administrative data

Description of key information

Acute toxicity via oral route

LD50 >5000 mg/kg/bw in female Wistar rats.

Acute toxicity via dermal route

LD50 >2000 mg/kg bw in male/female Wistar rats.

Acute toxicity via inhalation route

LD50 >5000 mg/m3 bw in male/female Wistar rats.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 June 2015 to 12 October 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

yes

Remarks:

Deviations from the maximum level of daily mean relative humidity occurred. Evaluation: Laboratory historical data do not indicate an effect of the deviations. The study integrity was not adversely affected by the deviation.

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

yes

Remarks:

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

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: 6 Females (nulliparous and non-pregnant). Each dose group consisted of 3 animals.Age and body weight: Young adult animals (approx. 10-11 weeks old) were selected. Body weight variation did not exceed +/- 20% of the sex mean.Identification: Earmark and tail markHealth inspection: At least prior to dosing. It was ensured that the animals were healthy and without any abnormality that might affect the study integrity.ConditionsEnvironmental controls for the animal room are 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 is between 07:00 and 19:00 hrs daily. The light/dark cycle may be interrupted for study related activities. Any variations to these conditions will be evaluated and maintained in the raw data.Accommodation: Group housing of 3 animals per cage in labeled Makrolon cages (MIV 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.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.

Route of administration:

oral: gavage

Vehicle:

propylene glycol

Details on oral exposure:

Method: Oral gavage, using plastic feeding tubes. The test item preparations were stirred on a magnetic stirrer during dosing.Fasting: Animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test substance. Water was available ad libitum.Frequency: Single dosage on Day 1.Dose level (volume): 2000 mg/kg (10 mL/kg) body weight.

Doses:

Single dose adminstered by oral gavage on day 1

No. of animals per sex per dose:

6 Females (nulliparous and non-pregnant). Each dose group consisted of 3 animals each given 2000 mg/kg (10 mL/kg) body weight.

Control animals:

not specified

Details on study design:

The toxicity of the test substance was assessed by stepwise treatment of groups of 3 females. The first group was treated at a dose level of 2000 mg/kg. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups.

Statistics:

The oral LD50 value of the test substance was ranked within the following ranges: 0-5, 5-50, 50-300 or 300-2000 mg/kg b.w. or as exceeding 2000 mg/kg b.w. The LD50 cut-off value was established based on OECD guideline 423. No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).The results were evaluated according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (including all amendments) and 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 (including all amendments).

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortaility occurs

Clinical signs:

Hunched posture and/or uncoordinated movements were noted for all animals on day 1

Body weight:

The body weight gain shown by the animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.

Gross pathology:

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

Mortality data

TEST DAY	1	1	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16
HOURS AFTER TREATMENT	0	2	4
FEMALES 2000 MG/KG	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
FEMALES 2000 MG/KG	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-

Clinical signs

TEST DAY	Max grade	1	1	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16
HOURS AFTER TREATMENT		0	2	4
FEMALES 2000 MG/KG Animal 1
Gait / motility. Uncoordinated movements	(3)	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Skin / fur. Piloerection	(1)	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Animal 2
Gait / motility. Uncoordinated movements	(3)	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Skin / fur. Piloerection	(1)	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Animal 3
Gait / motility. Uncoordinated movements	(3)	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Skin / fur. Piloerection	(1)	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Animal 4
Posture: Hunched posture	(1)	1	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Animal 5
Posture: Hunched posture	(1)	1	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Animal 6
Posture: Hunched posture	(1)	1	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-

- Sign not observed

Body weight gain

Sex/Dose level	ANIMAL	DAY 1	DAY 8	DAY 15
FEMALES 2000 MG/KG
	1	186	206	218
	2	177	191	205
	3	174	204	211
	MEAN	179	200	211
	ST.DEV.	6	8	7
	N	3	3	3
	1	193	212	221
	2	187	209	215
	3	199	216	226
	MEAN	193	212	221
	ST.DEV.	6	4	6
	N	3	3	3

Interpretation of results:

GHS criteria not met

Conclusions:

The oral LD50 value of Formaldehyde, oligomeric reaction products with acetone and diphenylamine in Wistar rats was established to exceed 5000 mg/kg body weight.

Executive summary:

The aim of the test was to assess the toxicity of the test substance when administered orally in a single dose to female rats. The test was conducted in accordance to:

Organization for Economic Co-operation and Development (OECD), OECD Guidelines for Testing of Chemicals, Section 4, Health Effects. No. 423, "Acute Oral Toxicity - Acute Toxic Class Method", 2001.

Commission Regulation (EC) No 440/2008 Part B: Methods for the Determination of Toxicity and other Health Effects; B1 tris: "Acute Oral Toxicity, Acute Toxic Class Method". Official Journal of the European Union No. L142, May 2008, including the most recent amendments.

United States Environmental Protection Agency (EPA). Health Effects Test Guidelines, OPPTS 870.1100, Acute Oral Toxicity. Office of Prevention, Pesticides and Toxic Substances (7101), EPA 712-C-02-190, 2002.

Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000; including the most recent partial revisions.

In conclusion the test concluded that the oral LD50 value for Formaldehyde, oligomeric reaction products with acetone and diphenylamine was established to exceed 2000 mg/kg/bw. No mortalities occurred, however, there were clinical signs such as hunched posture, piloerection and/or uncoordinated movements were noted for all animals on Day 1. The body weight gain shown by the animals was considered to be similar to that expected for normal untreated animals of the same age and stain. No abnormalities were found at macroscopic post mortem examination of the animals.

According to the OECD 423 test guideline, the LD50 cut-off value was considered to exceed 5000 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 oral toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2011) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments).

Endpoint conclusion

Endpoint conclusion:: no adverse effect observed
Dose descriptor:: LD50
Value:: 5 000 mg/kg bw
Quality of whole database:: K1 - GLP accredited laboratory study in accordance with recognised guidelines

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

01 August 2016 to 23 August 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

Organisation for Economic Co-operation and Development (OECD), OECD Guidelines for Testing of Chemicals, Section 4, Health Effects. No.403, "Acute Inhalation Toxicity", September 2009.

Deviations:

Qualifier:

according to guideline

Guideline:

EU Method B.2 (Acute Toxicity (Inhalation))

Version / remarks:

Commission Regulation (EC) No 440/2008 Part B: Methods for the Determination of Toxicity and other Health Effects; B.2. AcuteToxicity (inhalation). Official Journal of the European Communities No. L142, May 2008, including most recent amendments.

Deviations:

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Version / remarks:

EPA Health Effects Test Guidelines OPPTS 870.1300, Acute inhalation Toxicity. EPA 712-C-98-193, August 1998.

Deviations:

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Notification No 8147.

Version / remarks:

Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000; including the most recent partial revisions.

Deviations:

GLP compliance:

yes

Test type:

traditional method

Limit test:

yes

Specific details on test material used for the study:

Test item: 206534/B
Identification: Formaldehyde, oligomeric reaction products with acetone and diphenylamine
Appearance: Dark brown flakes
Batch: IC6D11P001
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: 27 April 2020 (expiry date)
Chemical name (IUPAC), synonym or trade name: Formaldehyde, oligomeric reaction products with acetone and diphenylamine (BXA)
CAS Number: 9003-80-9

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Species: Rat: Crl:WI(Han) (outbred, SPF-Quality). Recognised 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 nonpregnant) per exposure level. Two exposure levels
Age and body weight: Young adult animals were selected (approximately 10-11 weeks old). Animals used within the study were of approximately the same age and body weight variation did not exceed +/- 20% of the sex mean.
Identification: Individual unique number by tattoo on hind leg.
Health inspection: At least prior to exposure. It was ensured that the animals were healthy and without any abnormality that might affect the study integrity.

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.
Any variations to these conditions were maintained in the raw data and had no effect on the outcome of the study.
Accommodation
Group housing of five animals per sex per cage in labelled Makrolon cages (type IV; height 18 cm) containing sterilised sawdust as bedding material (Lignocel S 8-15, JRS -J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cageenrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
Acclimatisation period was at least 5 days before start of treatment under laboratory conditions.
Diet
Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) except during exposure to the test item.
Water
Free access to tap water except during exposure to the test item.

Animal husbandry on the day of exposure
The animals were moved to the inhalation area to in order to perform the exposure. During the exposure, there was no access to food and water. After exposure, the animals were returned to their cages which were placed in a fume cupboard for a short time period to allow test item remnants to evaporate. A sheet of filter paper was used to cover the bedding material to prevent suffocation in case of bad health condition and in order to recover and to aid the clinical observations. The sheet was removed and before the end of the exposure day, the surviving animals were returned to the animal room.
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.

Route of administration:

inhalation: dust

Type of inhalation exposure:

nose only

Vehicle:

air

Mass median aerodynamic diameter (MMAD):

>= 3.9 - <= 4.3 µm

Geometric standard deviation (GSD):

>= 1.9 - <= 2.1

Remark on MMAD/GSD:

The particle size distribution was characterized twice during each exposure period. The samples were drawn (2 L/min) from the test atmosphere through a tube mounted in one of the free animal ports of the middle section of the exposure chamber. The samples were collected with an 8 stage Marple personal cascade impactor containing fiber glass filters (TE-290-GF. Tisch Environmental, Cleves, Ohio, USA) and a fiber glass back-up filter (SEC-290-F1, Westech, Upper Stondon, Bedfordshire, England). Amounts of test item collected were measured gravimetrically. Subsequently the Mass Median Aerodynamic Diameter (MMAD) and the Geometric Standard Deviation (GSD) were determined.

Details on inhalation exposure:

Exposure Chamber
The design of the exposure chamber is based on the flow past nose-only inhalation chamber (Am. Ind. Hyg Assoc. J. 44(12): 923-928, 1983). The chamber (volume approximately 150 mL) consisted of three animal sections with eight animal ports each.
Each animal port had its own atmosphere inlet and exhaust outlet. The animals were placed in restraining tubes and connected to the animal ports. The number of animal sections and number of open inlets were adapted to the air flow in such a way that at each animal port the theoretical air flow was at least 1 L/min, which ensures an adequate oxygen supply to the animals. The main inlet of the test atmosphere was located at the top section and the main outlet was located at the bottom section. The direction of the flow of the test atmosphere guaranteed a freshly generated atmosphere for each individual animal.
All components of the exposure chamber in contact with the test material were made of stainless steel, glass, rubber or plastic. To avoid exposure of the personnel and contamination of the laboratory the exposure chamber was placed in a fume hood, which maintained at a slight negative pressure.

Test Atmosphere Generation
The test atmosphere generation was based on the method developed during extensive trial generations.
Administering the test item to a stream of pressurized air using a combination of a brush feeder and micronizing jet mill generated (Bernstein, D.N., Aerosols, pp 721- 723, 1984) generated a dust. The dust was passed through a series of three cyclones, allowing larger particles to settle, and diluted with pressurized air before it entered the exposure chamber. For the exposure at 1 mg/L, the mean total air flow was 23 L/min. and at 5 mg/L the mean total airflow was 20 L/min.
From the exposure chamber the test atmosphere was passed through a filter before it was released to the exhaust of the fume hood.

Test Atmosphere Characterization
Nominal Concentration
The nominal concentration was calculated by dividing the amount of test item used by the volume of pressurized air (average air flow times exposure time) entering the exposure chamber used for exposure of the animals. Due to the small volume of the exposure chamber the equilibrium time was negligible. The volume of air was calculated from the average air flow (measured by means of thermal mass flow meters and was recorded regularly, preferably in 30 minute intervals) and the exposure time.

Actual Concentration
A total of 21 and 28 representative samples were taken for determination of the actual concentrations during exposure at 5 and 1 mg/L, respectively. Samples were drawn from the test atmosphere through a tube mounted in one of the free animal ports of the middle section of the exposure chamber. Samples were drawn through a glass fiber filter (type APFC04700, Millipore, Billerica, MA, USA). The collected amount of the test item in the air sample was measured gravimetrically. Sample volumes were measured by means of a dry gas meter (type G 1.6, Actaris Meterfabriek B.V., Dordrecht, The Netherlands). Subsequently the timeweighted mean concentrations with the standard deviations were calculated.

Analytical verification of test atmosphere concentrations:

Duration of exposure:

4 h

Concentrations:

The study was performed following a stepwise exposure scenario. Target concentrations were based on the cut off concentration values specified in the UN and EC classification guidelines for dusts and mists (0.05, 0.5, 1 or 5 mg/L.).

No. of animals per sex per dose:

Toxicity data of the test item indicated that severe effects may be expected at the highest target concentration as indicated above. Therefore, as a first step the study was initiated with the exposure of five animals of each sex to 1 mg/L for 4 hours. Based on the results, five animals of each sex were exposed to 5 mg/L

Control animals:

Details on study design:

Stability Monitoring
It was considered that the opacity of the test atmosphere could not be reliably monitored by means of an aerosol monitoring system. An indication of the stability of the test atmosphere was obtained from the concentration measurements, which were equally distributed over time.

Temperature and Relative Humidity
The temperature and relative humidity were measured with a humidity and temperature indicator (E+E Elektronik, Engerwitzdorf, Austria) and were recorded after the animals were placed in the experimental set-up and at 30 minute intervals after initiation of the exposure.
The probe was inserted in a tube mounted in one of the free animal ports of the middle section of the exposure chamber.
The temperature of the atmosphere during the exposures were between 21.1 and 22.8°C and relative humidity was between 16 and 39%. These conditions were considered appropriate for this relatively short 4 hours exposure duration.

Treatment
Prior to each exposure, both eyes of each rat were instilled with Opthosan (AST Farma BV, Oudewater, The Netherlands) to protect the eyes against potential irritation by the test item.
Prior to exposure the animals were restrained in polycarbonate restraining tubes; these tubes were connected to the exposure chamber. Sixteen or eighteen minutes after the last animal was placed the generation of the test atmosphere was started. The exposure time was 4 hours.

Observations
Mortality/Viability: Twice daily.
Clinical signs: During exposure
Three times during exposure for mortality, behavioural signs of distress and effects on respiration.
Clinical signs: After exposure
On Day 1, one and three hours after exposure and once daily thereafter until Day 15. The clinical signs were graded according to fixed scales and the time of onset, degree and duration were recorded:
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).
Body weights: Days 1 (pre-administration), 2, 4, 8 and 15.
Necropsy: All animals were sacrificed at the end of the observation period by an intraperitoneal injection with Euthasol® (AST Farma BV, Oudewater, The Netherlands). All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities were recorded. Particular attention was given to any changes in the respiratory tract.

Statistics:

No statistical analysis was performed.

Preliminary study:

For the 1 mg/L exposure group, the time-weighted mean actual concentration was 1.1 ± 0.03 mg/L.

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5 mg/L air (nominal)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

No mortality occurred.

Clinical signs:

other: At 1 mg/L, quick breathing was seen during exposure. After exposure, one male showed hunched posture on Day 1. Brown staining by test item remnants of the skin and fur was seen between Days 1 and 6. At 5 mg/L, quick breathing was seen during exposure. Aft

Body weight:

Overall body weight gain in males and females was within the range expected for rats of this strain and age used in this type of study and were therefore considered not indicative of toxicity.

Gross pathology:

Macroscopic post mortem examination revealed several purple foci on the left side of the thymus of one female exposed to 1 mg/L. Macroscopic examination of the other animals did not reveal further abnormalities.

Other findings:

Concentration
For the 1 mg/L exposure group, the time-weighted mean actual concentration was 1.1 ± 0.03 mg/L. The nominal concentration (amount of test item used divided by the volume of pressurized air used) was 9.7 mg/L. The generation efficiency (ratio of actual and nominal concentration) was 11%.
The concentration was measured at time points (n=28) that were equally distributed over the exposure period, the results of which demonstrated that it was difficult to maintain the test item stable. The generation was interrupted on two occasions in order to make adjustments to the system. To compensate for these interruptions, the generation time was elongated by 9 minutes in order to achieve an actual exposure time of 240 minutes. By calculating the time weighted mean concentration, the effect of the interruptions and variations were taken into account. Overall, it was considered that the generation represented the intended exposure to the 1 mg/L target concentration for 4 hours.
For the 5 mg/L exposure group, the time-weighted mean actual concentration was 5.3 ± 0.4 mg/L. The nominal concentration was 64.1 mg/L and the generation efficiency was 8%.
The concentration was measured at time points (n=21) that were equally distributed over the exposure period, the results of which showed that it was difficult to maintain a stable concentration. The generation was interrupted twice and the set-up was changed within the first 50 minutes after start. The generation time was elongated with 12 minutes in order to achieve an actual exposure time of 240 minutes. By calculating the time weighted mean concentration, the effect of the interruptions and variations were taken into account. Overall, it was considered that the generation represented the intended exposure to the 5 mg/L target concentration for 4 hours.

Particle Size
The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice during the exposure period. At 1 mg/L, the MMAD was 3.9 μm (gsd 1.9) and 4.0 μm (gsd 2.1). At 5 mg/L, the MMAD was 4.0 μm (gsd 2.1) and 4.3 μm (gsd 2.0).

Gravimetric Concentration of Test Item

1 mg/L exposure group

Start of generation of test atmosphere: 09:55

End of generation of test atmosphere: 14:04

Time

(hh:mm)

Action

Sample volume

(L)

Mass sampled

(mg)

Concentration

(mg/L)

% of total exposure time

Weight concentration

(mg/L)

9:55

9:59

10:03

10:10

10:18

10:24

10:31

10:48

11:02

11:08

11:14

11:20

11:32

11:51

12:09

12:24

12:31

12:37

12:43

12:49

12:54

13:00

13:03

13:10

13:11

13:17

13:23

13:28

13:34

13:38

13:40

13:42

13:59

14:04

Start of generation

Start of sampling

Stop generation

Start of generation

Start of sampling

Stop generation

Start of generation

Start of sampling

End of generation

n.a.

3.90

4.62

4.02

4.84

6.34

5.40

6.07

7.45

8.60

8.08

5.91

5.02

5.97

6.71

6.98

7.53

7.62

4.45

3.52

1.91

0.83

n.a.

8.34

8.80

4.34

4.85

4.09

n.a.

5.55

4.48

n.a.

0.780

0.924

0.804

0.968

1.057

1.080

1.214

1.490

1.720

1.616

1.182

1.004

1.194

1.342

1.396

1.506

1.524

0.890

0.704

0.382

0.166

0.166¹⁾

n.a.

1.668

1.760

0.868

0.970

0.818

0.818¹⁾

n.a.

1.110

0.896

0.896¹⁾

0.0

1.7

2.9

3.3

2.5

2.9

7.1

5.8

2.5

5.0

7.9

7.5

6.3

2.9

2.5

2.1

2.5

1.3

n.a.

0.4

2.5

2.1

2.5

1.7

n.a.

0.8

7.1

2.1

n.a.

0.013

0.015

0.023

0.032

0.026

0.032

0.086

0.087

0.043

0.040

0.030

0.050

0.095

0.101

0.087

0.044

0.038

0.022

0.018

0.008

0.004

0.002

n.a.

0.007

0.044

0.022

0.020

0.014

n.a.

0.009

0.063

0.019

Time-weighted mean concentration

Standard deviation

Number of samples

1.115

0.028

1) Assumed concentration, based on the last sample

2) n.a. = not applicable

Gravimetrical Concentration of Test Item

5 mg/L exposure group

Start of generation of test atmosphere: 09:14

End of generation of test atmosphere: 13:26

Time

(hh:mm)

Action

Sample volume

(L)

Mass sampled

(mg)

Concentration

(mg/L)

% of total exposure time

Weight concentration

(mg/L)

9:14

9:17

9:22

9:28

9:31

9:34

9:35

9:42

9:48

9:58

10:07

10:08

10:11

10:19

10:21

10:29

10:35

10:41

10:46

10:54

11:16

11:20

11:37

11:59

12:21

13:20

13:26

Start of generation

Start of sampling

Stop generation

Start of generation

Start of sampling

Stop generation

Start of generation

Start of sampling

End of generation

n.a.

12.74

8.56

3.98

n.a.

14.91

1.00

0.49

n.a.

41.86

5.51

79.38

40.47

8.27

33.03

39.39

9.42

38.13

10.64

36.97

32.38

39.08

37.05

49.66

n.a.

2.548

1.712

0.796

0.796¹⁾

n.a.

2.982

0.200

0.082

0.082¹⁾

n.a.

8.372

1.102

15.876

8.094

1.654

6.606

7.878

1.884

7.626

2.1228

7.394

6.476

7.816

7.410

6.208

6.208¹⁾

0.0

1.3

2.1

2.5

1.3

n.a.

0.4

2.9

2.5

4.2

n.a.

0.4

1.2

3.3

0.8

3.3

2.5

2.1

3.3

9.2

1.7

7.1

9.2

24.6

2.5

n.a.

0.032

0.036

0.020

0.010

n.a.

0.012

0.006

0.002

0.003

n.a.

0.035

0.014

0.529

0.067

0.055

0.165

0.197

0.039

0.254

0.195

0.123

0.459

0.716

0.679

1.526

0.155

Time-weighted mean concentration

Standard deviation

Number of samples

5.331

0.351

1) Assumed concentration, based on the last sample

2) n.a. = not applicable

Aerodynamic Particle Size Distribution in the Test Atmosphere

1 mg/L exposure group

Start of generation of test atmosphere: 09:55

End of generation of test atmosphere: 14:04

Sampling speed (L/min): 2

Measurement 1:

Sampling time: 10:56

Sample volume (L): 8

Stage

Cut point

(μm)

Mass sampled

(mg)

Relative mass

(%)

Cumulative mass

(% of total sampled)

Back up

21.0

15.0

10.0

6.0

3.5

2.0

0.9

0.5

0.25

0.01

0.07

1.00

1.89

3.25

1.32

0.48

0.15

0.14

0.12

0.84

12.03

22.74

39.11

15.88

5.78

1.81

1.68

99.88

99.04

87.00

64.26

25.15

9.27

3.49

1.68

0.00

MMAD¹

(μm):

gsd²:

3.9

1.9

¹Mass Median Aerodynamic Diameter

²Geometric standard deviation

Measurement 2:

Sampling time: 13:47

Sample volume (L): 9

Stage

Cut point

(μm)

Mass sampled

(mg)

Relative mass

(%)

Cumulative mass

(% of total sampled)

Back up

21.0

15.0

10.0

6.0

3.5

2.0

0.9

0.5

0.25

0.00

0.07

1.23

1.66

2.34

1.17

0.38

0.09

0.26

0.00

0.97

17.08

23.06

32.50

16.25

5.28

1.25

3.61

100.00

99.03

81.94

58.89

26.39

10.14

4.86

3.61

0.00

MMAD¹

(μm):

gsd²:

4.0

2.1

¹Mass Median Aerodynamic Diameter

²Geometric standard deviation

Aerodynamic Particle Size Distribution in the Test Atmosphere

5 mg/L exposure group

Start of generation of test atmosphere: 09:14

End of generation of test atmosphere: 13:26

Sampling speed (L/min): 2

Measurement 1:

Sampling time: 11:54

Sample volume (L): 2

Stage

Cut point

(μm)

Mass sampled

(mg)

Relative mass

(%)

Cumulative mass

(% of total sampled)

Back up

21.0

15.0

10.0

6.0

3.5

2.0

0.9

0.5

0.25

0.00

0.24

0.48

0.47

0.66

0.26

0.06

0.00

11.06

22.12

21.66

30.41

11.98

2.76

0.00

100.00

88.94

66.82

45.16

14.75

2.76

0.00

MMAD¹

(μm):

gsd²:

4.0

2.1

¹Mass Median Aerodynamic Diameter

²Geometric standard deviation

Measurement 2:

Sampling time: 13:24

Sample volume (L): 2

Stage

Cut point

(μm)

Mass sampled

(mg)

Relative mass

(%)

Cumulative mass

(% of total sampled)

Back up

21.0

15.0

10.0

6.0

3.5

2.0

0.9

0.5

0.25

0.00

0.84

0.95

1.76

0.79

0.72

0.14

0.01

0.00

16.12

18.23

33.78

15.16

13.82

2.69

0.19

100.00

83.88

65.64

31.86

16.70

2.88

0.19

0.00

MMAD¹

(μm):

gsd²:

4.3

2.0

¹Mass Median Aerodynamic Diameter

²Geometric standard deviation

Mortality

TEST DAY

HOURS AFTER TREATMENT

MALES 1 MG/LITER

FEMALES 1 MG/LITER

MALES 5 MG/LITER

FEMALES 5 MG/LITER

Clinical Signs

TEST DAY

HOURS AFTER TREATMENT

MAX

GRADE

MALES 1 MG/LITER

ANIMAL 1

Skin / fur

Brown staining (Ear left)

Brown staining (Ear right)

Brown staining (General)

(1)

ANIMAL 2

Skin / fur

Brown staining (Ear left)

Brown staining (Ear right)

Brown staining (General)

(1)

ANIMAL 3

Posture

Hunched posture

Skin / fur

Brown staining (Ear left)

Brown staining (Ear right)

Brown staining (General)

(1)

ANIMAL 4

Skin / fur

Brown staining (Ear left)

Brown staining (Ear right)

Brown staining (General)

(1)

ANIMAL 5

Skin / fur

Brown staining (Ear left)

Brown staining (Ear right)

Brown staining (General)

(1)

FEMALES 1 MG/LITER

ANIMAL 6

Skin / fur

Brown staining (Ear left)

Brown staining (Ear right)

Brown staining (General)

(1)

ANIMAL 7

Skin / fur

Brown staining (Ear left)

Brown staining (Ear right)

Brown staining (General)

(1)

ANIMAL 8

Skin / fur

Brown staining (Ear left)

Brown staining (Ear right)

Brown staining (General)

(1)

ANIMAL 9

Skin / fur

Brown staining (Ear left)

Brown staining (Ear right)

Brown staining (General)

(1)

ANIMAL 10

Skin / fur

Brown staining (Ear left)

Brown staining (Ear right)

Brown staining (General)

(1)

MALES 5 MG/LITER

ANIMAL 11

Skin / fur

Green staining (General)

Brown staining (Ears)

(1)

ANIMAL 12

Skin / fur

Green staining (General)

Brown staining (Ears)

(1)

ANIMAL 13

Skin / fur

Green staining (General)

Brown staining (Ear right)

Brown staining (Ears)

(1)

ANIMAL 14

Skin / fur

Green staining (General)

Brown staining (Ear right)

Brown staining (Ears)

(1)

ANIMAL 15

Skin / fur

Green staining (General)

Brown staining (Ears)

(1)

FEMALES 5 MG/LITER

ANIMAL 16

Skin / fur

Green staining (General)

Brown staining (Ears)

(1)

ANIMAL 17

Skin / fur

Green staining (General)

Brown staining (Ears)

(1)

ANIMAL 18

Skin / fur

Green staining (General)

Brown staining (Ear right)

Brown staining (Ears)

(1)

ANIMAL 19

Skin / fur

Green staining (General)

Brown staining (Ears)

(1)

ANIMAL 20

Skin / fur

Green staining (General)

Brown staining (Ears)

(1)

Body Weights (Gram)

SEX/DOSE LEVEL

ANIMAL

DAY 1

DAY 2

DAY 4

DAY 8

DAY 15

MALES 1 MG/LITER

MEAN

ST.DEV.

299

334

318

313

326

318

294

328

316

307

320

313

300

336

321

314

325

319

312

350

332

320

335

330

331

376

356

340

360

353

FEMALES 1 MG/LITER

MEAN

ST.DEV.

195

198

196

200

214

201

194

200

201

209

211

203

191

198

208

211

213

204

194

202

207

209

215

205

216

219

221

220

224

220

MALES 5 MG/LITER

MEAN

ST.DEV.

279

285

276

290

264

279

266

264

263

276

252

264

277

276

273

289

259

275

291

297

292

308

273

292

315

319

313

335

290

314

FEMALES 5 MG/LITER

MEAN

ST.DEV.

184

206

187

181

194

190

178

195

184

180

186

185

182

195

188

179

196

188

193

204

199

185

209

198

207

212

202

215

207

Macroscopic Findings

ANIMAL

ORGAN

FINDING

DAY OF DEATH

MALES 1 MG/LITER

No findings noted

Scheduled necropsy

Day 15 after treatment

Scheduled necropsy

Day 15 after treatment

Scheduled necropsy

Day 15 after treatment

Scheduled necropsy

Day 15 after treatment

Scheduled necropsy

Day 15 after treatment

FEMALES 1 MG/LITER

Thymus

No findings noted

Left side: focus/foci, several, purple

No findings noted

Scheduled necropsy

Day 15 after treatment

Scheduled necropsy

Day 15 after treatment

Scheduled necropsy

Day 15 after treatment

Scheduled necropsy

Day 15 after treatment

Scheduled necropsy

Day 15 after treatment

MALES 5 MG/LITER

No findings noted

Scheduled necropsy

Day 15 after treatment

Scheduled necropsy

Day 15 after treatment

Scheduled necropsy

Day 15 after treatment

Scheduled necropsy

Day 15 after treatment

Scheduled necropsy

Day 15 after treatment

FEMALES 5 MG/LITER

No findings noted

Scheduled necropsy

Day 15 after treatment

Scheduled necropsy

Day 15 after treatment

Scheduled necropsy

Day 15 after treatment

Scheduled necropsy

Day 15 after treatment

Scheduled necropsy

Day 15 after treatment

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results, the inhalatory LC50, 4h value of Formaldehyde, oligomeric reaction products with acetone and diphenylamine in Wistar rats was considered to exceed 5 mg/L.

Executive summary:

Assessment of acute inhalation toxicity with Formaldehyde, oligomeric reaction products with acetone and diphenylamine in the rat (nose-only)

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

OECD Guidelines, Section 4, Health Effects. No.403, "Acute Inhalation Toxicity", Sep 2009.

Commission Regulation (EC) No 440/2008, B.2. Acute Toxicity (inhalation), L142, May 2008.

EPA OPPTS 870.1300, Acute inhalation Toxicity. EPA 712-C-98-193, August 1998.

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

Formaldehyde, oligomeric reaction products with acetone and diphenylamine was administered as a dust by nose-only inhalation for 4 hours to two groups of five male and five female Wistar rats. Mortality and clinical signs were observed daily during the observation period and body weights were determined on Days 1, 2, 4, 8 and 15. Macroscopic examination was performed after terminal sacrifice (Day 15).

For the 1 mg/L exposure group, the time-weighted mean actual concentration was 1.1 ± 0.03 mg/L. The nominal concentration (amount of test item used divided by the volume of pressurized air used) was 9.7 mg/L. The generation efficiency (ratio of actual and nominal concentration) was 11%. The concentration was measured at time points (n=28) that were equally distributed over the exposure period, the results of which demonstrated that it was difficult to maintain the test item stable. Overall, it was considered that the generation represented the intended exposure to the 1 mg/L target concentration for 4 hours.

For the 5 mg/L exposure group, the time-weighted mean actual concentration was 5.3 ± 0.4 mg/L. The nominal concentration was 64.1 mg/L and the generation efficiency was 8%. The concentration was measured at time points (n=21) that were equally distributed over the exposure period, the results of which showed that it was difficult to maintain a stable concentration. Overall, it was considered that the generation represented the intended exposure to the 5 mg/L target concentration for 4 hours.

The Mass Median Aerodynamic Diameter (MMAD) and geometric standard deviation (gsd) were determined twice during the exposure period. At 1 mg/L, the MMAD was 3.9 μm (gsd 1.9) and 4.0 μm (gsd 2.1). At 5 mg/L, the MMAD was 4.0 μm (gsd 2.1) and 4.3 μm (gsd 2.0).

No mortality occurred.

At 1 mg/L, quick breathing was seen during exposure. After exposure, one male showed hunched posture on Day 1. Brown staining by test item remnants of the skin and fur was seen between Days 1 and 6.

At 5 mg/L, quick breathing was seen during exposure. After exposure, no systemic clinical signs were seen. Brown and green staining by test item remnants of the skin and fur was seen between Days 1 and 4.

Overall body weight gain in males and females was within the range expected for rats of this strain and age used in this type of study and were therefore considered not indicative of toxicity.

Based on the results, the inhalatory LC50, 4h value of Formaldehyde, oligomeric reaction products with acetone and diphenylamine in Wistar rats was considered to exceed 5 mg/L.

Based on this study, Formaldehyde, oligomeric reaction products with acetone and diphenylamine does not have to be classified and has no obligatory labelling requirement for acute inhalation 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.

Endpoint conclusion

Endpoint conclusion:: no adverse effect observed
Dose descriptor:: LC50
Value:: 5 000 mg/m³
Quality of whole database:: Klimisch 1 study

Acute toxicity: via dermal route

Link to relevant study records

Reference

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

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"

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"

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

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)

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.

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

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

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

HOURS AFTER TREATMENT

MAX GRADE

MALES 2000 MG/KG

ANIMAL 1

Posture

Flat posture

Various

Ptosis

Black (Treated skin)

(1)

(3)

(1)

ANIMAL 2

Posture

Flat posture

Various

Ptosis

Black (Treated skin)

(1)

(3)

(1)

ANIMAL 3

Posture

Flat posture

Skin / fur

Scales (Treated skin)

Various

Ptosis

Black (Treated skin)

(1)

(3)

(1)

ANIMAL 4

Posture

Flat posture

Skin / fur

Scales (Treated skin)

Scabs

Secretion / excretion

Chromodacryorrhoea (Snout)

Various

Ptosis

Black (Treated skin)

(1)

(3)

(1)

ANIMAL 5

Posture

Flat posture

Skin / Fur

Scales (Treated skin)

Various

Ptosis

Black (Treated skin)

(1)

(3)

(1)

FEMALES 2000 MG/KG

ANIMAL 6

Posture

Flat posture

Secretion / excretion

Chromodacryorrhoea (Snout)

Various

Ptosis

Black (Treated skin)

(1)

(3)

(1)

ANIMAL 7

Posture

Flat posture

Various

Ptosis

Black (Treated skin)

(1)

(3)

(1)

ANIMAL 8

Posture

Flat posture

Various

Ptosis

Black (Treated skin)

(1)

(3)

(1)

ANIMAL 9

Posture

Flat posture

Skin / fur

Scales (Treated skin)

Scabs

Various

Ptosis

Black (Treated skin)

(1)

(3)

(1)

ANIMAL 10

Posture

Flat posture

Various

Ptosis

Black (Treated skin)

(1)

(3)

(1)

- = sign not observed

BODY WEIGHTS (GRAM)

SEX/DOSE LEVEL

ANIMAL

DAY 1

DAY 8

DAY 15

MALES 2000 MG/KG

276

286

276

277

284

286

291

287

278

300

303

314

306

291

327

MEAN

ST. DEV.

280

288

308

FEMALES 2000 MG/KG

182

189

186

181

198

186

194

188

195

206

192

210

191

MEAN

ST. DEV.

186

189

199

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

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).

Endpoint conclusion

Endpoint conclusion:: no adverse effect observed
Dose descriptor:: LD50
Value:: 2 000 mg/kg bw
Quality of whole database:: K1 - GLP accredited laboratory study in accordance with recognised guidelines

Additional information

Acute toxicity via oral route

The aim of the test was to assess the toxicity of the test substance when administered orally in a single dose to female rats. The test was conducted in accordance with OECD Guideline 423. The oral LD50 value for Formaldehyde, oligomeric reaction products with acetone and diphenylamine was established to exceed 5000 mg/kg/bw. No mortalities occurred, however, there were clinical signs such as hunched posture, piloerection and/or uncoordinated movements were noted for all animals on Day 1. The body weight gain shown by the animals was considered to be similar to that expected for normal untreated animals of the same age and stain. No abnormalities were found at macroscopic post mortem examination of the animals.

Acute toxicity via dermal route

The aim of the test was to assess the toxicity of the test substance when administered by the dermal route to rats. The test was conducted in accordance with OECD Guideline 402. 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.

Acute toxicity via inhalation route

The aim of the test was to assess the toxicity of the test substance when administered by the inhalation route (nose-only) to rats. The test was conducted in accordance with OECD Guideline 403. Formaldehyde, oligomeric reaction products with acetone and diphenylamine was administered as a dust by nose-only inhalation for 4 hours to two groups of five male and five female Wistar rats. Mortality and clinical signs were observed daily during the observation period and body weights were determined on Days 1, 2, 4, 8 and 15. Macroscopic examination was performed after terminal sacrifice (Day 15).

No mortality occurred. At 1 mg/L, quick breathing was seen during exposure. After exposure, one male showed hunched posture on Day 1. Brown staining by test item remnants of the skin and fur was seen between Days 1 and 6. At 5 mg/L, quick breathing was seen during exposure. After exposure, no systemic clinical signs were seen. Brown and green staining by test item remnants of the skin and fur was seen between Days 1 and 4. Overall body weight gain in males and females was within the range expected for rats of this strain and age used in this type of study and were therefore considered not indicative of toxicity. Macroscopic post mortem examination revealed several purple foci on the left side of the thymus of one female exposed to 1 mg/L. Macroscopic examination of the other animals did not reveal further abnormalities.

The generation of high dust concentrations is technically challenging. In this study, the concentration varied considerably during the first 2 hours. Over the total 4 hour exposure period, the time weighted mean concentration exceeded 5 mg/L. Considering the absence of mortality and severe signs, no significant mortality would be expected after exposure to 5 mg/L with less variation. Based on these results, the inhalatory LC50, 4h value of Formaldehyde, oligomeric reaction products with acetone and diphenylamine in Wistar rats was considered to exceed 5 mg/L.

Justification for classification or non-classification

Acute toxicity via oral route

According to the OECD 423 test guideline, the LD50 cut-off value was considered to exceed 5000 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 oral toxicity according to Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments).

Acute toxicity via dermal route

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 Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).

Acute toxicity via inhalation route

Based on the results, the inhalatory LC50, 4h value of Formaldehyde, oligomeric reaction products with acetone and diphenylamine in Wistar rats was considered to exceed 5 mg/L. Based on the results of this study, formaldehyde, oligomeric reaction products with acetone and diphenylamine does not have to be classified and has no obligatory labelling requirement for acute inhalation toxicity according to Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments.

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