Benzenesulfonic acid, 4-chloro-2-[2-[4,5-dihydro-3-methyl-5-oxo-1-(3-sulfophenyl)-1H-pyrazol-4-yl]diazenyl]-5-methyl-, ammonium salt (1:2)

Endpoint summary

Description of key information

In an oral acute toxicity study (GLP) according to EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure), no mortality or adverse treatment-related effects were observed in male and female rats. The median lethal oral dose to rats was found to be greater than 2000 mg/kg bodyweight.

In a dermal acute toxicity study (GLP) according to EU Method B.3 (Acute Toxicity (Dermal)), no mortality or adverse treatment-related effects were observed in male and female rats. No irritation or other dermal changes were observed. The median lethal dermal dose to rats was found to be greater than 2000 mg/kg bodyweight.

Experimental data of a read across substance are used to evaluate the acute inhalative toxicity of the test substance. A single administration of the analogue substance via respiratory system did not cause health effects or mortalities in rats. The LC50 is > 5.5 mg/L air.

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:

May - June 1994

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Lot/batch number of test material: KL384
- Expiration date of the lot/batch: not specified
- Purity test date: not specified

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature in the dark
- Stability under storage conditions: not specified
- Stability under test conditions: not specified
- Solubility and stability of the test substance in the solvent/dispersant/vehicle/test medium: not specified

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: suspended in distilled water
- Final dilution of a dissolved solid, stock liquid or gel: 20 % concentration in distilled water

FORM AS APPLIED IN THE TEST (if different from that of starting material) : suspended in distilled water. The test substance was prepared on the day of dosing.

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Olac Ltd., Bicester, Oxon, England
- Age at study initiation: approximately 4 to 7 weeks
- Weight at study initiation: males: 102.8 g (mean); females: 96.2 g (mean)
- Fasting period before study: Access to food only was prevented overnight prior to and approximately 4 hours after dosing.
- Housing: up to 5 rats of the same sex per cage
- Diet: standard laboratory rodent diet ad libitum
- Water: drinking water ad libitum
- Acclimation period: 5 days
- Microbiological status when known : not specified
- Method of randomisation in assigning animals to test and control groups: not specified

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 - 24
- Humidity (%): 56
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: To: not specified

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 20%

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg bodyweight.

Doses:

2 g/kg bodyweight

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: Animals were observed soon after dosing and at frequent intervals for the remainder of Day 1 (a period of five hours). On subsequent days animals were observed once in the morning and again at the end of the experimental day. The bodyweight of each rat was recorded on Days 1 (prior to dosing), 2,3,4, 8 and 15.
- Necropsy of survivors performed: All animals were subjected to a macroscopic examination which consisted of opening the abdominal and thoracic cavities. The macroscopic appearance of all tissues was recorded.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality and no signs of specific toxicity observed

Mortality:

none

Clinical signs:

- Piloerection was observed in all rats within five minutes of dosing and throughout the remainder of day 1
- Yellow discolouration of the faeces was evident for all animals on day 2 only

Body weight:

All rats achieved anticipated bodyweight gains throughout the study.

Gross pathology:

No macroscopic abnormalities were observed.

Interpretation of results:

GHS criteria not met

Conclusions:

The acute lethal oral dose to rats was found to be greater than 2.0 g/kg bodyweight.

Executive summary:

A group of ten fasted rats (five males and five females) was given a single dose by gavage of the test substance, formulated in distilled water, at a dose level of 2.0 g/kg bodyweight. All animals were killed and examined macroscopically on Day 15, the end of the observation period.

There were no deaths. Clinical signs of reaction to treatment were limited to piloerection, recovery was complete by Day 2. However, a yellow discolouration of the faeces was evident for all animals on Day 2 only. All rats achieved the anticipated bodyweight gains throughout the study. No abnormalities were recorded at the macroscopic examination on Day 15.

The acute lethal oral dose to rats was found to be greater than 2.0 g/kg bodyweight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

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:

Nov 1981

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

limited data to test substance

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

yes

Remarks:

No analytical purity; gravimetric analyses of the concentration without analytical confirmation of the composition and reanalysis of the airborne material, respectively; single instead of twice particle size distribution determination.

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: WIGA Versuchstierzuchtanstalt, Sulzfeld
- Age at study initiation: ca. 8 - 10 weeks
- Weight at study initiation: 307 +/- 15 g (males), 228 +/- 17 g (females)
- Housing: 5 animals/cage
- Diet: SSNIFF R Alleindiät für Ratten und Mäuse (SSNIFF-Versuchstierdiäten GmbH, Soest) ad libitum
- Water: tap water ad libitum
- Acclimation period: not specified
- Method of randomisation in assigning animals to test and control groups: not specified

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2
- Humidity (%): 55 +/- 5
- Air changes (per hr): not specified
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: To: not specified

Route of administration:

other: dust aerosol

Type of inhalation exposure:

nose/head only

Vehicle:

air

Mass median aerodynamic diameter (MMAD):

1.2 µm

Geometric standard deviation (GSD):

4

Details on inhalation exposure:

Head-nose inhalation system INA 20 (glass/steel construction, BASF Aktiengesellschaft, V ca. 55 l); the animals are restrained in tubes and their snouts project into the inhalation chamber.
A mixture of dust and air was generated by means of a vibration aerosol generator (fluidized bed).
By means of a dust generator the substance to be tested was generated into a dust aerosol, which was passed into the inhalation system.
A vibration aerosol generator was used for generating dust. The concentration was adjusted by varying the amplitude and frequency of the vibrator.
The rate of flow was adjusted as follows: 1500 l/h of compressed air through the vibrator.
The inhalation mixture was offered to the animals for inhalation for 4 hours.
By means of an exhaust air system the pressure ratios in the inhalation chamber were adjusted in such a way that the amount of fresh air was about 10% lower. This ensured that the mixture of test substance and air was not diluted by laboratory air in the breathing zones of the animals.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

5.5 mg/l

No. of animals per sex per dose:

10

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: before starting the study, after 7 days and after the end of the observation period
- Necropsy of survivors performed: yes

Statistics:

The statistical evaluation of the concentration-response relationship was carried out in accordance with the binominal test (Wittig, H.: Mathematische Statistik 1974, pp. 32 - 35) according to tables of the testing facility.
The particle size was determined in accordance with mathematical and graphical methods of evaluating particle measurements (Silverman, L.: Particle Size Analysis in Industrial Hygiene, 1971, pp. 235-259).

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.5 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

none

Clinical signs:

other: During exposure: substance-colored fur in the region of the head. After exposure: substance-colored fur, partly agitated behavior, after 1 day nothing abnormal detected.

Body weight:

The body weight of the animals showed no adverse effects in comparison with that of the control.

Gross pathology:

Sacrificed animals: no abnormalities detected.

Mean body weight:

Test animals; before start of the study: 307 g (males), 228 g (females); after 7 days: 341 g (males), 241 g (females); after 14 days: 370 g (males), 252 g (females)

Control animals: before start of the study 307 g (males), 230 g (females); after 7 days 340 g (males), 240 g (females); after 14 days 370 g (males), 249 g (females)

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating conc.

Value:

5 500 mg/m³

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:

June 1994

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Lot/batch number of test material: KL384
- Expiration date of the lot/batch: not specified
- Purity test date: not specified


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature in the dark
- Stability under storage conditions: not specified
- Stability under test conditions: not specified
- Solubility and stability of the test substance in the solvent/dispersant/vehicle/test medium: not specified

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: suspended in distilled water
- Final dilution of a dissolved solid, stock liquid or gel: 71.43 % concentration in distilled water

FORM AS APPLIED IN THE TEST (if different from that of starting material): suspended in distilled water. The test substance was prepared on the day of dosing.

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Olac Ltd., Bicester, Oxon, England
- Age at study initiation: approximately 7 to 10 weeks
- Weight at study initiation: males: 240.8 g (mean); females: 226.2 g (mean)
- Fasting period before study: no
- Housing: one animal per cage
- Diet: standard laboratory rodent diet ad libitum
- Water: drinking water ad libitum
- Acclimation period: 6 days
- Microbiological status when known: not specified
- Method of randomisation in assigning animals to test and control groups: not specified

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 - 24
- Humidity (%): 52
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: To: not specified

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: dorso-lumbar region
- % coverage: approximately 10% of the total body surface
- Type of wrap if used: The treated area (approximately 50 mm x 50 mm) was covered with gauze which was held in place with a non-irritative dressing encircled firmly around the trunk.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated area of skin was washed with warm (30° to 40°C) water and blotted dry with absorbent paper.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2.0 g/kg bodyweight
- Constant volume or concentration used: yes
- For solids, paste formed: not specified. The suspended test substance was applied by spreading it evenly over the prepared skin.

Duration of exposure:

24 h

Doses:

2 g/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: Animals were observed soon after dosing and at frequent intervals for the remainder of Day 1 (a period of three hours). On subsequent days animals were observed once in the morning and again at the end of the experimental day. Local dermal irritation at the treatment site was assessed daily. Individual bodyweights were recorded on Days 1 (prior to dosing), 8 and 15.
- Necropsy of survivors performed: All animals were killed on Day 15 by cervical dislocation and were subjected to a macroscopic examination which consisted of opening the abdominal and thoracic cavities. The macroscopic appearance of all examined tissues was recorded.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Remarks on result:

other: no mortality and no signs of specific toxicity observed

Mortality:

none

Clinical signs:

There were no signs of systemic reaction to treatment.

Body weight:

- A slightly low bodyweight gain was recorded for one male and one female on Day 8 and in three males on Day 15. All other rats achieved the anticipated bodyweight gains throughout the study.

Gross pathology:

No macroscopic abnormalities were observed.

Other findings:

Sites of application of the test substance showed no irritation or other dermal changes. However, a residual yellow staining was evident in a number of animals during the study clearing in all instances by Day 15.

Interpretation of results:

GHS criteria not met

Conclusions:

The acute lethal dermal dose to rats was found to be greater than 2.0 g/kg bodyweight.

Executive summary:

A group of ten rats (five males and five females) was given a single dermal application of the test substance, at a maximum practical concentration of 71.43% w/v in distilled water, and at a dose level of 2.0 g/kg bodyweight. All animals were killed and examined macroscopically on Day 15, the end of the observation period. There were no deaths and no signs of systemic reaction to treatment. Sites of application of the test item showed no irritation or other dermal changes. However, a residual yellow staining was evident in a number of animals during the clearing in all instances by Day 15. A slightly low bodyweight gain was recorded for one male and one female on Day 8 and in three males on Day 15. All other rats achieved the anticipated bodyweight gains throughout the study. No abnormalities were recorded at the macroscopic examination on Day 15.

The acute lethal dermal dose to rats was found to be greater than 2.0 g/kg bodyweight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Additional information

Acute oral toxicity:

In an oral acute toxicity study (GLP) according to EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure), a group of ten fasted rats (five males and five females) was given a single dose by gavage of the test substance, formulated in distilled water, at a dose level of 2.0 g/kg bodyweight. All animals were killed and examined macroscopically on Day 15, the end of the observation period.

There were no deaths. Clinical signs of reaction to treatment were limited to piloerection, recovery was complete by Day 2. However, a yellow discolouration of the faeces was evident for all animals on Day 2 only. All rats achieved the anticipated bodyweight gains throughout the study. No abnormalities were recorded at the macroscopic examination on Day 15.

The acute lethal oral dose to rats was found to be greater than 2000 mg/kg bodyweight.

Acute dermal toxicity:

In a dermal acute toxicity study (GLP) according to EU Method B.3 (Acute Toxicity (Dermal)), a group of ten rats (five males and five females) was given a single dermal application of the test substance, at a maximum practical concentration of 71.43% w/v in distilled water, and at a dose level of 2.0 g/kg bodyweight. All animals were killed and examined macroscopically on Day 15, the end of the observation period.

There were no deaths and no signs of systemic reaction to treatment. Sites of application of the test item showed no irritation or other dermal changes. However, a residual yellow staining was evident in a number of animals during the clearing in all instances by Day 15. A slightly low bodyweight gain was recorded for one male and one female on Day 8 and in three males on Day 15. All other rats achieved the anticipated bodyweight gains throughout the study. No abnormalities were recorded at the macroscopic examination on Day 15.

The acute lethal dermal dose to rats was found to be greater than 2000 mg/kg bodyweight.

Acute inhalative toxicity:

The test substance was not analyzed for its acute toxicity by inhalation. Experimental data of a structural analogue are available. The substances are salts of sulphonatophenyl-pyrazol-azobenzenesulfonic acid and differ in their respective inorganic cation and two substituents only. A detailed read across justification is attached in chapter 7.2.2 of the IUCLID dossier.

In a non-GLP study equivalent to OECD guidelline 403, male and female rats (10/sex/dose) were exposed for 4 h to a dust aerosol (5.5 mg/L air, 1.500 L/h, nose only) of the analogue substance. Following dosing, the animals were observed for 14 d. Examination of clinical signs and viability were performed daily. There were no deaths. Clinical signs or changes in body weight gain were not observed. Gross necropsy was without any findings. Thus, the LC50 was greater than 5.5 mg/L air, which is also 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. The oral and dermal LD50 in rats was greater than 2000 mg/kg bw. The LC50 of an analogue substance was greater than 5.5 mg/L air. No mortality was observed in each of these studies. As a result the actual substance is not considered to be classified for acute toxicity under Regulation (EC) No. 1272/2008.