4,4'-[isopropylidenebis(4,1-phenyleneoxy)]dianiline

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1985

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

/ Basic data given.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

other information

Study period:

1985

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

No information available about the tested concentration.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Inhalation of substantially saturated vapor, single exposure for 6 hours, static conditions 25 °C

GLP compliance:

no

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Remarks:

Albino

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 200 - 300 g

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 25

Route of administration:

inhalation: vapour

Type of inhalation exposure:

not specified

Vehicle:

not specified

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: The vapour was produced by enclosing the test substance in a sealed animal chamber for 18 h (static conditions). Oxygen was added as needed in order to maintain a chamber oxygen content of approximatively 20%
- Exposure chamber volume: 120 L
- Temperature in air chamber: 25 °C

Analytical verification of test atmosphere concentrations:

not specified

Duration of exposure:

6 h

Concentrations:

substantially saturated vapor

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Weighing was performed in the beginning and on Day 7 and 14
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Sex:

male/female

Dose descriptor:

LC50

Based on:

test mat.

Exp. duration:

6 h

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

No mortality was observed during the observation period.

Clinical signs:

other: No clinical signs were observed.

Body weight:

All animals showed normal body weight gains during the observation period. (in males: mean body weight gain of 68 g and 120 g after 7 and 14 days, respectively; in females: mean body weight gain of 18 g and 29 g after 7 and 14 days, respectively)

Gross pathology:

No gross pathologic findings were observed.

Interpretation of results:

study cannot be used for classification

Cross-reference 2

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1985

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

/ Basic data given.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

adopted in 2017

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2 - 3 kg
- Housing: Rabbits were immobilized during 24 h contact period

Type of coverage:

occlusive

Vehicle:

corn oil

Details on dermal exposure:

TEST SITE
- Type of wrap if used: The test material was retained under impervious sheeting on the clipped, intact skin of the trunk.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Excess fluid was removed to diminish ingestion.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied: not reported
- For solids, paste formed: yes, test substance was moistened with corn oil

Duration of exposure:

24 h

Doses:

4 and 8 g/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Remarks:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observations of the skin were performed at 1 h, on Day 7 and 14, animals were weighed on Day 7 and 14
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 8 000 mg/kg bw

Based on:

test mat.

Mortality:

There was no mortality during the study period.

Clinical signs:

The high dose administration (8 g/kg bw) resulted in erythema (Day 1) in all 10 animals (female/males), edema (Day 7) in 2 males and 2 females. Desquamation was apparent on the dosed skin from Day 7 till Day 14 in 2 males and in all 5 females. Administration of the lower dose (4 g/kg bw) produced erythema (Day 1) and desquamation (Day 7 - 14) in all 10 animals (females/males).

Body weight:

The high dose administration (8 g/kg bw) resulted in emaciation (Day 7) in 2 males and 1 female, who recovered at Day 14, while in 1 female it was observed at Day 14. Administration of the lower dose (4 g/kg bw) had no impact on the body weight of any animal during the study period.

Gross pathology:

Gross pathologic examination revealed slightly mottled and dark red lungs in 2 males and enlarged kidneys and spleens in 2 females after administration of the high dose (8 g/kg bw), while administration of the lower dose (4 g/kg bw) produced no gross pathologic findings.

Table 1: Body weight changes

	Dose [mg/ kg bw]	Body weight change
		Day 7 (range (mean) [g] )	Day 14 (range (mean) [g] )
Male	8000	-597 to -289 (-381)	-430 to -182 (-294)
	4000	-566 to -130 (-352)	-344 to -57 (-191)
Female	8000	-606 to -2819 (-370)	-632 to -57 (-317)
	4000	-466 to -247 (-348)	-264 to -32 (-158)

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008

Conclusions:

CLP: not classified

Cross-reference 3

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1985

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

/ Basic data given.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted in 2015

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

other: The test substance was moistened with corn oil.

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.5 g

Duration of treatment / exposure:

4 h

Observation period:

7 days

Number of animals:

3 males/ 3 females

Details on study design:

TEST SITE
- Area of exposure: 2.45 x 2.45 cm² on dorsal skin
- Type of wrap if used: The dose is applied to the clipped, intact skin under a gauze patch and is loosely covered with impervious sheeting.

REMOVAL OF TEST SUBSTANCE
- Washing: Excess test substance was removed after contact.
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS
1, 24, 48, 72 h and 7 days

SCORING SYSTEM:
- Method of calculation: Draize scoring system

Irritation parameter:

erythema score

Basis:

mean

Remarks:

of 6 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritation parameter:

edema score

Basis:

mean

Remarks:

of 6 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: reversibility: not applicable

Irritant / corrosive response data:

No skin reaction was observed at any of the test sites of the 6 animals throughout the observation period.

Other effects:

Mortality/Clinical signs:
No mortality or clinical signs of toxicity were observed until the end of the observation period.
Sample residue was observed on 1/6 rabbits at 5 h.

Interpretation of results:

other: CLP/ EU GHS criteria not met, classification not required according to Regulation (EC) No 1272/2008

Conclusions:

CLP: not classified

Cross-reference 4

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1985

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

/ Basic data given.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted in 2017

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.1 g (approx. 0.1 mL)

Duration of treatment / exposure:

single application without washing

Observation period (in vivo):

7 days
Reading time points: 1, 4, 24, 48, 72 h and Day 7

Number of animals or in vitro replicates:

3 males/ 3 females

Details on study design:

SCORING SYSTEM:
Cornea:
(A) Opacity - degree of density (most dense area taken for reading)
No opacity: 0
Scattered or diffuse area, iris details clearly visible: 1
Easily discernible translucent areas, iris details slightly obscured: 2
Opalescent areas, iris details not visible. pupil size barely discernible: 3
Opaque, iris invisible: 4

(A) Area of cornea involved
One-quarter or less, but not zero: 1
Greater than one-quarter, but less than half: 2
Greater than half, but less than three-quarter: 3
Greater than three-quarter, up to whole area: 4

Iris:
(A)
Normal: 0
Folds above normal, congestion, swelling, circumcorneal injection (any or all), iris still reacting to light: 1
No reaction to light. hemorrhage, gross destruction: 2

Conjunctivae:
(A)
Vessels normal: 0
Vessels definitely injected above normal: 1
Diffuse, deep crimson red, individual vessels not readily discernible: 2
Diffuse beefy red: 3

(B)
No chemosis: 0
Any swelling above normal (includes nicitating membrane): 1
Obvious swelling with partial eversion of lids: 2
Swelling with lids about half closed: 3
Swelling with lids about half closed to completely closed: 4
(C)
No discharge: 0
Any amount of discharge different from normal: 1
Discharge with moistening of the lid, and hairs adjacent to lids: 2
Discharge with considerable moistening around the eyes: 3

TOOL USED TO ASSESS SCORE: Fluorescein sodium solution (2%) was used to determine corneal injury before dosing and at readings after one day.

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of 6 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 6 animals

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

of 6 animals

Time point:

24/48/72 h

Score:

0.06

Max. score:

3

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: 1 female showed conjunctivae (redness) scored 1 at 24 h

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 6 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritant / corrosive response data:

1 h after the application the test substance produced iritis (scored 1) in 1/6 animals persisting until the 24 h reading timepoint and slight conjunctival irritation in 4/6 animals (redness and chemosis, scored 1). The redness was then observed in all 6 animals at the 4 h reading time point but was reversible in 5/6 animals within 24 h, while for 1/6 animals (female) this was reversible at 48 h.
Discharge was observed 1 h after application of the test substance in all 6 animals. This reaction was reversible in 1 and 5 animals at 4 h and 24 h, respectively.
No corneal effects were apparent in any animal and time point.

Table 1: Individual scores of eye irritation

Rabbit #	Time [h]	Conjunctivae		Iris	Cornea
		Redness	Chemosis (swelling)
1 (m)	1	1	1	0	0
	4	1	0	0	0
	24	0	0	0	0
	48	0	0	0	0
	72	0	0	0	0
	mean	0	0	0	0
2 (m)	1	1	1	0	0
	4	1	0	0	0
	24	0	0	0	0
	48	0	0	0	0
	72	0	0	0	0
	mean	0	0	0	0
3 (m)	1	0	0	0	0
	4	1	0	0	0
	24	0	0	0	0
	48	0	0	0	0
	72	0	0	0	0
	mean	0	0	0	0
4 (f)	1	0	0	0	0
	4	1	0	0	0
	24	0	0	0	0
	48	0	0	0	0
	72	0	0	0	0
	mean	0	0	0	0
5 (f)	1	1	1	1	0
	4	1	1	1	0
	24	1	0	0	0
	48	0	0	0	0
	72	0	0	0	0
	mean	0.33	0	0	0
6 (f)	1	1	1	0	0
	4	1	0	0	0
	24	0	0	0	0
	48	0	0	0	0
	72	0	0	0	0
	mean	0	0	0	0

Only the values from the reading time points 24, 48 and 72 h were used to calculate the mean values.

Interpretation of results:

other: CLP/ EU GHS criteria not met, classification not required according to Regulation (EC) No 1272/2008

Conclusions:

CLP: not classified

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1985

Report date:

1985

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

4,4'-[isopropylidenebis(4,1-phenyleneoxy)]dianiline

EC Number:

235-985-8

EC Name:

4,4'-[isopropylidenebis(4,1-phenyleneoxy)]dianiline

Cas Number:

13080-86-9

Molecular formula:

C27H26N2O2

IUPAC Name:

4,4'-[isopropylidenebis(4,1-phenyleneoxy)]dianiline

Test animals

Species:

rat

Strain:

Sprague-Dawley

Remarks:

Albino

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation:
- Weight at study initiation: 200 - 300 g
- Fasting period before study: animals were fasted overnight
- Diet: approprtiate commercial diet, ad libitum
- Water: municipal water, ad libitum

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 10% (w/v)
- Justification for choice of vehicle: The substance was not suspending in water

Doses:

125, 250, 500 and 1000 mg/kg bw (males)
125, 250 and 500 mg/kg bw (females)

No. of animals per sex per dose:

5

Control animals:

no

Remarks:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Day 0, Day 7 and Day 14 (prior to sacrifice)
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross pathologic evaluation

Results and discussion

Effect levels

Effect levels 1

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

308 mg/kg bw

Based on:

test mat.

Remarks on result:

other: LD50 values are calculated by the moving average method (Thompson 1947) and are based on a 14-day observation period.

Effect levels 2

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

406 mg/kg bw

Based on:

test mat.

Remarks on result:

other: LD50 values are calculated by the moving average method (Thompson 1947) and are based on a 14-day observation period.

Mortality:

females: 3/5 animals died on Day 5, 1/5 animals on Day 6 and 1/5 animals on Day 8 at a concentration of 500 mg/kg bw; 1/5 animals died on Day 5 at 250 mg/kg bw and no animal died at 125 mg/kg bw.

males: 1/5 animals died on Day 3, 1/5 animals on Day 4, 2/5 animals on Day 5 and 1/5 animals on Day 6 at a concentration of 1000 mg/kg bw; 1/5 animals died on Day 5, 1/5 animals on Day 8 and 1/5 animals on Day 14 at a concentration of 500 mg/kg bw; 1/5 animals died on Day 6 at 250 mg/kg bw and no animal died at 125 mg/kg bw.

Clinical signs:

females: At a test substance concentration of 500 mg/kg/bw sluggishness, unkempt appearance, red crust on perinasal and periocular fur was observed at Day 4, swollen eyelids and kyphosis at Day 5. At a test substance concentration of 250 mg/kg/bw sluggishness, unkempt appearance, red crust on perinasal and periocular fur was observed at Day 3, emaciation and intermittent tremors in 1 animal at Day 7 and alopecia and unkempt appearance at Day 14. No clinical signs were observed at the lowest test substance concentration.

males: At a test substance concentration of 1000 mg/kg/bw sluggishness and yellow periurogenital discharge was observed at Day 1, unkempt appearance in 2 animals at Day 1, swollen eyelids (eyes partially closed), red perinasal and periocular discharge and kyphosis at Day 3, intermittent tremors and emaciation at Day 4. At a test substance concentration of 500 mg/kg/bw red perinasal crust was observed at Day 2, sluggishness, swollen eyelids, kyphosis and red periocular crust at Day 3, unkempt appearance and emaciation at Day 4, lacrimation in 2 animals at Day 5, intermittent tremors in 2 animals at Day 7 alopecia in 1 animal at Day 5. Recovery of 1 survivor at Day 9. At a test substance concentration of 250 mg/kg/bw red perinasal crust was observed at Day 2, sluggishness and swollen eyelids at Day 3, unkempt appearance and emaciation at Day 4. At a test substance concentration of 125 mg/kg bw unkempt appearance was observed in 2 animals at Day 5. Recovery was observed at Day 7.

Body weight:

All animals showed normal body weight gains during the observation period.

Gross pathology:

females: Lungs mottled light and dark red at a test substance concentration of 500 mg/kg bw. At 250 mg/kg bw, 1/5 females (the one which died later) showed red encrustation of perinasal and periocular fur while in the other 4 animals nothing remarkable was observed. At 125 mg/kg bw no gross pathological abnormalities were observed.
males: Lungs mottled light and dark red at a test substance concentration of 1000 mg/kg bw. At 500 mg/kg bw lungs of 3/5 females (the ones which died later) were dark red and the thoracic and abdominal cavity of 1 animal was filled with brown liquid. In the other 2 animals, nothing remarkable was observed. At 250 mg/kg bw, 1/5 males (the one which died later) showed nothing remarkable, while the cervical lymph nodes of 2/5 animals were enlarged. At 125 mg/kg bw, the spleen of 1/5 animals was enlarged.

Any other information on results incl. tables

Table 1: Mortality and body weight changes

	Dose [mg/ kg bw]	Dead/ Dosed animals	Days of death	Body weight change
				Day 7 (range (mean) [g] )	Day 14 (range (mean) [g] )
Male	1000	5/5	3, 4, 5, 5, 6	-	-
	500	3/5	5, 8, 14	-38 - 9 (-18)	-2 - 42 (20)
	250	1/5	6	2 - 19 (8)	61 - 92 (78)
	125	0/5	3, 4, 5, 5, 6	30 - 49 (38)	87 - 120 (111)
Female	500	5/5	5, 5, 5, 6, 8	173	-
	250	1/5	5	2 - 19 (10)	22 - 58 (38)
	125	0/5	-	-10 - 33 (5)	40 - 60 (47)

Applicant's summary and conclusion

Interpretation of results:

other: CLP/ EU GHS criteria met, classification required according to Regulation (EC) No 1272/2008

Conclusions:

CLP: Acute oral toxicity, Cat. 4, H302