Benzoic acid, 2,3,4,5-tetrachloro-6-cyano-, methyl ester, reaction products with p-phenylenediamine and sodium methoxide

Endpoint summary

Description of key information

- Oral: similar to OECD guideline 401, prior GLP, LD50 (rat) >5000 mg/kg bw (no mortality or signs of toxicity)

- Inhalation: similar to OECD guideline 403, prior GLP, LC50 (rat) >1.04 mg/L (no mortality or signs of toxicity)

- Dermal: read across to CAS 30125-47-4, similar to OECD guideline 402, prior GLP, LD50 (rat) >2500 mg/kg bw (no mortality or signs of toxicity)

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:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

unknown concentration in vehicle, uncommon dose selection, high dose volume in PEG

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Specific details on test material used for the study:

- Physical state: solid
- Analytical purity: no data

Species:

rat

Strain:

other: Tif: RAif (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 7 to 8 weeks
- Mean weight at study initiation per treatment group: males 193-208 g (maximum SD: 13.4 g); females 172-188 g (maximum SD: 9.4)
- Fasting period before study: overnight
- Housing: groups of 5 in Macrolon cages (type 3)
- Diet (e.g. ad libitum): NAFAG, Gossau SG, CH; ad lib.
- Water (e.g. ad libitum): not specified; ad lib.
- Acclimation period: at least 4 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+-2
- Humidity (%):55+-10
- Photoperiod (hrs dark / hrs light): 10/14

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Details on oral exposure:

VEHICLE
- Concentration in vehicle: no data

MAXIMUM DOSE VOLUME APPLIED: 10 and 20 mL/kg bw

Doses:

2000, 3000, 4000 and 5000 mg/kg bw (5000 mg/kg bw was reported to be the highest possible dose)

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: Physical condition and rate of deaths were monitored throughout the whole observation period. Bodyweights were recorded immediately prior to dosing (control weights) and at 7 and 14 days.
- Necropsy of survivors performed: yes. The animals were submitted at random to a necropsy at the end of the observation period.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality observed.

Mortality:

No mortality observed.

Clinical signs:

other: Dyspnoea, exophthalmus, curved position and ruffled fur were observed in all treatment groups and were reversible within at least 8 days (2000-3000 mg treatment) or 9 days (>=4000 mg).

Gross pathology:

No substance-related gross organ changes were seen.

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

5 000 mg/kg bw

Quality of whole database:

Similiar to OECD 401, pre-GLP, Klimisch 2

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

no data on purity

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

Principles of method if other than guideline:

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:

traditional method

Limit test:

yes

Specific details on test material used for the study:

- Physical state: solid
- Analytical purity: no data

Species:

rat

Strain:

other: Tif:RAIf (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 185-190 g
- Housing: 9 per cage (Macrolon, Type 4)
- Diet (e.g. ad libitum): NAFAG, Gossau SG, CH; ad lib.
- Water (e.g. ad libitum): unspecified; ad lib.
- Acclimation period: at least 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:

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. The concentration and the particle size distribution of the dust in the vicinity of the animals were 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.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

1.04 +/- 0.07 mg/L ( original data: 1041 +/- 69 mg/m^3)

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: observation intervals were not reported; weighing was not performed
- Necropsy of survivors performed: yes

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 1.04 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: No mortality and no clinical signs were observed.

Mortality:

No mortality observed

Clinical signs:

other: No signs of toxicity observed

Body weight:

Not measured

Gross pathology:

No substance related gross organ changes were seen.

Interpretation of results:

GHS criteria not met

Executive summary:

No mortality and clinical signs were observed at the highest technically attainable concentration.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating conc.

Value:

1 041 mg/m³ air

Physical form:

inhalation: dust / mist

Quality of whole database:

Similiar to OECD 403, pre-GLP, Klimisch 2

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

please refer to the attached read-across justification

Reason / purpose for cross-reference:

read-across source

Statistics:

not performed

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 500 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality and no clinical signs were observed.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 500 mg/kg bw

Quality of whole database:

Read across to CAS 30125-47-4, similiar to OECD 402, pre-GLP, Klimisch 2

Additional information

Oral:

In the course of the key study, acute oral toxicity was examined by single oral (gavage) administration of the test material dissolved in polyethylene glycol at doses of 2000, 3000, 4000 or 5000 mg/kg bw to male and female Tif: RAif rats (5/sex/dose, fasted overnight). Bodyweights were recorded immediately prior to dosing (control weights) and at 7 and 14 days. Physical condition and rate of deaths were monitored throughout the whole observation period. All animals survived until the scheduled day of necropsy, no substance related gross organ changes were seen. Therefore, the LD50 was >5000 mg/kg bw.

 

Next to the key study mentioned above, several other acute oral toxicity studies in rats and mice were performed. All of the animals received single doses of the test substance up to 5000 or 7000 mg/kg bw, respectively, and were observed for a minimum of 8 days. Mortality did not occur in any of the studies; clinical signs or organ changes were also not observed. In two of the studies dyspnoe, curved position and ruffled fur were recorded; these symptoms were reversible within the post observation period.

 

Inhalation:

Acute inhalation toxicity was examined by a 4h dust exposure (1000 mg/m³ (highest technically attainable conc., nose only) to 9 male and female Tif: RAif rats. The exposure was started 15 minutes after onset of the dust production, when the dust had reached an even dispersal throughout the chamber. After a 4 hour inhalation, the rats were returned to their cages. Physical conditions and incidence of death were monitored throughout an observation period of 14 days. None of the animals died, organ changes or clinical signs of toxicity were not observed. The LC50 was considered to be greater than 1.04 mg/L air.

 

Dermal:

Acute toxicity after dermal application of the test item was not evaluated. However, experimental data of a structural analogue (CAS 30125-47-4) are available.

To determine acute dermal toxicity of an analogue substance, 2500 mg/kg bw of the material was applied onto dorsal skin of male and female rats for 24h under occlusive conditions. The animals were observed for 14 days and checked daily for mortality and clinical signs of toxicity. None of the rats died; yellowish staining of the treatment site was observed. The LD50 was set >2500 mg/kg bw. 

Based on the physico-chemical, structural as well as toxicoligical similarities, the same outcome is assumed for the actual substance.

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 1272/2008. No mortality occurred. As a result, the substance is not considered to be classified for acute oral, inhalation or dermal toxicity under Regulation (EC) No. 1272/2008, as amended for the fourteenth time in Regulation (EC) No. 2020/217.