3,3'-[(2,5-dimethyl-p-phenylene)bis[imino(1-acetyl-2-oxoethylene)azo]]bis[4-chloro-N-(5-chloro-o-tolyl)benzamide]

Endpoint summary

Description of key information

Oral: LD50 > 5000 mg/kg bw/day, OECD 401

Dermal: LD50 > 5000 mg/kg bw/day and additional read-across (CAS 5580-57-4): LD50 > 5000 mg/kg bw/day

Inhalative (dust): Read-across (CAS 5580-57-4): LC50 (4h, rat) > 1.7 mg/L air (highest concentration possible), OECD 403

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

no

Remarks:

Quality Assurance Unit supervision

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

commercial grade

Species:

rat

Strain:

other: Tif : RAIf (SPF), F3-crosses of RII 1/Tif x RII 2/Tif

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Ciba-Geigy Ltd. Tierfarm, Sisseln / Switzerland
- Age at study initiation: 7 - 8 weeks
- Weight at study initiation: 177 - 217 g
- Fasting period before study: overnight
- Housing: in groups of five
- Diet: Rat food, NAFAG No. 890, NAFAG AG, Gossau / Switzerland; ad libitum
- Water: ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 55 +/- 15%
- Air changes (per hr): approximately 15
- Photoperiod (hrs dark / hrs light): 12 / 12

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

(PEG 400)

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 20 mL/kg bw

Doses:

5000 mg/kg bw

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: Mortality: daily; a.m. and p.m. on working days, a.m. on weekend days; Signs and Symptoms: daily; Body weight: on days 1, 7, 14 and at death
- Necropsy of survivors performed: yes

Statistics:

From the body weights, the group means and their standard deviations were calculated.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortalities observed.

Mortality:

No mortalities observed.

Clinical signs:

other: Dyspnoea (day 1 - day 10), exophthalmus (day1 - day 9), ruffled fur (day 1 - day 7), diarrhea (day 1) and curved body position (day 1 - day 5) were observed, being common symptoms in acute tests. In addition a transient sedation was noted. The surviving a

Gross pathology:

Besides some enlarged caeca (in 4 male and 2 female animals), no noticeable findings were made at necropsy.

Interpretation of results:

GHS criteria not met

Conclusions:

According to the test conditions the test substance has practically no acute toxicity when adminstered orally to the albino rat.

Executive summary:

In an acute oral toxicity study (BAF, 1983), groups of 5 male and 5 female Wistar rats were given a single oral dose of the test substance in polyethylene glycol at a dose of 5000  mg/kg bw and observed for 14 days. No animals died and the treated animals did showed common symptoms as dyspnoea, exophtalmus, ruffled fur and curved body position. In addition a transient sedation was noted. Therefore the LD50 is greater than 5000 mg/kg bw. In conclusion, according to the test conditions the test substance has practically no acute toxicity when adminstered orally to the albino rat.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: inhalation

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

READ ACROSS ANALOGUE APPROACH

The read-across hypothesis is fundamentally based on the same core structure of five ‘yellow disazo condensation pigments’ which optionally can serve as target or as source substances. None of the pigments are sufficiently soluble, either in water or in octanol for systemic uptake or metabolism. The molecular weight ranges from 716.6 g/mol (Pigment Yellow 155) to 1229.2 g/mol (Pigment Yellow 128). Therefore, the molecular weights of all ‘yellow disazo condensation pigments’ are well above the threshold of 500 g/mol, which is generally considered for low dermal and oral uptake [ECHA Guidance R. 7c, 2017]. Furthermore, for each of the substances, the critical body burden (CBB) is above the octanol solubility, which generally indicates a low uptake in biota and makes toxicity unlikely [ECHA Guidance R. 11, 2017].


Please find the complete Read-across justification text including a data matrix and structures attached in the attachment.

Reason / purpose for cross-reference:

read-across source

Principles of method if other than guideline:

Assessment based on read-across to study similar to OECD 403

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 1.7 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: No mortality and clinical signs observed

Reference 2

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

(no data on test substance purity, limited documentation)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

yes

Remarks:

(no data on body weight determination during the test; dark / light period 10/14 instead of 12/12)

Principles of method if other than guideline:

Inhalation toxicity was tested according to the method of Sachsse et al. (1973).
K. Sachsse, L. Ullmann, G. Voss and R. Hess: Measurement of inhalation toxicity of aerosols in small laboratory animals. In: Proceedings of the Europ. Soc. for the Study of Drug Toxicity. Vol. XV, pp. 239-251, Zurich, June 1973.

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

- Lot/batch No.: EN 42476.75

Species:

rat

Strain:

other: Tif : RAIf (SPF) strain

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: own breed
- Weight at study initiation: mean = 190 to 195 g
- Fasting period before study: no data
- Housing: 9 animals to a cage (the males and females were segregated); in Macrolon cages, type 4
- Diet: Rat food - NAFAG, Gossau SG, ad libitum
- Water: ad libitum
- Acclimation period: for a minimum of 4 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 1
- Humidity (%): 55 ± 5
- Photoperiod (hrs dark / hrs light): 10 / 14

Route of administration:

inhalation: dust

Type of inhalation exposure:

nose only

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION

The rats were kept on separate PVC tubes positioned radially around the exposure chamber such that snout and nostrils of the animals only were exposed to the dust. The exposure was started 15 minutes after onset of the dust production, when the dust had reached an even dispersal throughout the chamber. The dust was generated by injecting the test material with the help of a "Grafix Exaktomat Injector" into an air stream which was discharged into the exposure chamber through a nozzle under a pressure of 2 atm at a rate of 20 L/min.

TEST ATMOSPHERE
- Brief description of analytical method used: The concentration of the dust in the vicinity of the animals was monitored at 1 hour intervals throughout the dust exposure.
- Samples taken from breathing zone: yes

TEST ATMOSPHERE
- Particle size distribution:
The particle size distribution of the dust in the vicinity of the animals was monitored at 1 hour intervals throughout the dust exposure. The concentration was determined gravimetrically by sampling the test atmosphere through a selectron filter of 50 mm diameter and with a pore size of 0.2 µm (Schleicher and Schuell, Feldbach, Switzerland) at an air flow rate of 10 L/min. The size distribution of the dust particles was measured with a Cascade Impactor with selectron filters of 25 mm diameter and with a pore size of 0.2 µm (Schleicher and Schuell) at an air flow rate of 17.5 L/min. 5% have a particle size < 1 µm and 40% are between 3 - 7 µm, 25 - 30% are between 1 - 3 µm and additionally, 25 - 30% are > 7 µm.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

1692 ± 313 mg/m³ (Average dust concentration, no higher concentrations were possible.)

No. of animals per sex per dose:

9

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: no data
- Necropsy of survivors performed: yes

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 1.7 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: No mortality and clinical signs observed.

Mortality:

No mortality was observed.

Clinical signs:

other: During the 4-hour exposure and the subsequent 14-day observation period no toxic symptoms were observed.

Body weight:

No data.

Gross pathology:

No substance related gross organ changes were seen.

Interpretation of results:

GHS criteria not met

Conclusions:

The LC50 of a 4 hour dust exposure for rats of both sexes is greater than 1.7 mg/L.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

1 700 mg/m³ air

Acute toxicity: via dermal route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: Only short abstract available and original reference not translated (czech)

Principles of method if other than guideline:

Rats were applied dermally with the test substance and observed for 14 days.

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

commercial grade
purity not specified

Species:

rat

Strain:

Wistar

Sex:

male

Details on test animals or test system and environmental conditions:

- Weight at study initiation: 233-277 g

Type of coverage:

not specified

Vehicle:

olive oil

Duration of exposure:

no data

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

3

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: yes

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortalities observed.

Mortality:

No mortalities occured during observation period.

Clinical signs:

other: No clinical signs observed.

Gross pathology:

No macroscopic abnormalities observed.

Reference 2

Endpoint:

acute toxicity: dermal

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Justification for type of information:

READ ACROSS ANALOGUE APPROACH

The read-across hypothesis is fundamentally based on the same core structure of five ‘yellow disazo condensation pigments’ which optionally can serve as target or as source substances. None of the pigments are sufficiently soluble, either in water or in octanol for systemic uptake or metabolism. The molecular weight ranges from 716.6 g/mol (Pigment Yellow 155) to 1229.2 g/mol (Pigment Yellow 128). Therefore, the molecular weights of all ‘yellow disazo condensation pigments’ are well above the threshold of 500 g/mol, which is generally considered for low dermal and oral uptake [ECHA Guidance R. 7c, 2017]. Furthermore, for each of the substances, the critical body burden (CBB) is above the octanol solubility, which generally indicates a low uptake in biota and makes toxicity unlikely [ECHA Guidance R. 11, 2017].


Please find the complete Read-across justification text including a data matrix and structures attached in the attachment.

Reason / purpose for cross-reference:

read-across source

Principles of method if other than guideline:

Read-across assessment based on in vivo studies in rat

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortalities observed.

Reference 3

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: Only short abstract available and original reference not translated (czech)

Principles of method if other than guideline:

Rats were applied dermally with the test substance and observed for 14 days.

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male

Details on test animals or test system and environmental conditions:

- Weight at study initiation: 245-303 g

Type of coverage:

not specified

Vehicle:

olive oil

Duration of exposure:

no data

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

3

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: yes

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortalities observed.

Mortality:

No mortalities occured during observation period.

Clinical signs:

other: No clinical signs observed.

Gross pathology:

No macroscopic abnormalities observed.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 mg/kg bw

Additional information

Oral:

Reliable data from two studies on acute toxicity after oral application are available for the test substance. These data reveal a very low acute oral toxicity of the test substance: LD50 values in rats are above 5000 mg/kg bw, the upper limit for classification.

In an acute oral toxicity study (Ciba-Geigy Ltd., 1983), groups of 5 male and 5 female Wistar rats were given a single oral dose of the test substance in polyethylene glycol at a dose of 5000 mg/kg bw and observed for 14 days. No animals died and the treated animals showed common symptoms as dyspnoea, exophtalmus, ruffled fur and curved body position. In addition a transient sedation was noted. Therefore the LD50 is greater than 5000 mg/kg bw. In conclusion, according to the test conditions the test substance has practically no acute toxicity when administered orally to the albino rat.

Additionally a supporting study with the test substance is available performed with a much higher concentration (Sythesia, 1989). In this study groups of 5 male and 5 female Wistar rats were given a single oral dose of the test substance in olive oil at a dose of 15850  mg/kg bw and observed for 14 days. No animals died and the treated animals did not show any abnormalities. Therefore the LD50 is greater than 15850 mg/kg bw.

 

Dermal:

In two acute dermal toxicity studies (Synthesia, 1989), three male Wistar rats were dermally exposed to 5000 mg/kg bw test substance (CAS 5580-57-4 and 5280-80-8). Animals then were observed for 14 days. No mortality occurred. No systemic signs were observed in the animals during the entire observation period. No macroscopical organ findings were observed in the animals.

Although these studies are only short abstracts they can be used as weight of evidence since they show both the same results. Furthermore, the substances are also not irritant after skin contact and reveal a low log Pow which indicates that these substances are hardly absorbed through the skin. Therefore, no classification for acute dermal toxicity is necessary for the members of the 'yellow disazo condensation pigments'.

 

Inhalation:

In an acute inhalation toxicity study (similar to OECD 403, Ciba-Geigy Ltd., 1976), groups of Tif:RAIf rats (9/sex) were exposed to dust of the test substance (CAS 5580-57-4) for 4 hours and observed for 14 days. No mortality occurred during 14 day observation. At concentrations of 1700 mg/m³ air at the 4 hour exposure the animals showed no toxic symptoms. At autopsy, no deviations from normal morphology were found in all animals. 1700 mg/m³ air was the highest possible concentration. That leads to an LC50 greater than 1700 mg/m³ air at 4 hour exposure. As no lethal effects occurred at the maximum technically feasible concentration it is concluded that the members of the 'yellow disazo condensation pigments' have not to be classified for acute toxicity after inhalation exposure.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008
The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on available data on acute toxicity, the test item is not classified according to Regulation (EC) No 1272/2008 (CLP), as amended for the tenth time in Regulation (EU) No 2017/776.