Ammonium sodium 2-[4-[[1-[[(2-methoxy-5-methyl-4-sulphonatophenyl)amino]carbonyl]-2-oxopropyl]azo]phenyl]-6-methylbenzothiazole-7-sulphonate

Endpoint summary

Description of key information

Acute oral toxicity

The acute oral LD50 was determined to be > 2000 mg/kg bw.

Acute dermal toxicity

The acute dermal LD50 was determined to be > 2000 mg/kg bw.

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:

2015-12-16 to 2016-01-26

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)

Version / remarks:

December 17, 2001

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

2002

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japan MAFF 8147

Version / remarks:

2000

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Specific details on test material used for the study:

Batch identification: 13298465
Expiry date: January 2019
Storage conditions: Room temperature

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga GmbH, Germany
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: Young adult animals (female animals approx. 10 weeks)
- Weight at study initiation: Animals of comparable weight 175-189 g (± 20% of the mean weight group 1: 183.7 g, group 2: 177.7 g, actual weights)
- Fasting period before study: 16 hours before administration
- Housing: Single housing
- Diet: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany, ad libitum
- Water: Tap water ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22°C +/- 3°C
- Humidity: 30 – 70%
- Air changes per hr: Approx. 10
- Photoperiod (hrs dark / hrs light): 12 h / 12 h (6.00 a.m. – 6.00 p.m. / 6.00 p.m. – 6.00 a.m.)

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION: The test item was ground with mortar and pestle. The test item preparation was produced for each test group shortly before application by stirring with a high speed homogenizer (Ultra-Turrax) and a magnetic stirrer. Additionally, the homogeneity of the test item preparation during application was ensured by stirring with a magnetic stirrer.

CLASS METHOD
- Rationale for the selection of the starting dose: By request of the sponsor a starting dose of 2000 mg/kg bw was chosen in the first step with 3 female animals.
Because no mortality occurred, 2000 mg/kg bw were administered to another group of 3 female animals in the second step.

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:

Body weight determination: Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation.

Clinical observations: Clinical signs for each animal were recorded several times on the day of administration and at least once during each workday thereafter.

Mortality: A check for any dead or moribund animals was made at least once each workday; these records are archived by Bioassay.

Necropsy of survivors performed: yes
Pathology: Necropsy with gross-pathology examination was performed on the last day of the observation period after sacrifice by CO2-inhalation in a chamber with gradually increasing concentrations.

Histology: No histological examinations were performed.

Conducted analyses:
Stability of the test item preparation
Homogeneity of the test item preparation
Concentration control analysis of the test item preparation
Analysis of feed
Analysis of drinking water
Bedding and enrichment analysis

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred

Clinical signs:

No clinical signs were observed during clinical examination

Body weight:

The body weight of the animals increased within the normal range throughout the study period with one exception. In one animal the body weight increased normally during the first week, but this animal only slightly gained weight during the second week. This effect is observed at times in the rat strain used, because in the required age range the female animals have already reached the phase of slow growth.

Gross pathology:

There were no macroscopic pathological findings in all animals sacrificed at the end of the observation period.

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

GLP and guideline compliant study

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

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

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:

February 24, 1987

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Version / remarks:

August 1998

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japan MAFF 8147

Version / remarks:

2000

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

Batch identification: 13298465
Expiry date: January 2019
Storage conditions: Room temperature

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga GmbH, Germany
- Female animals were nulliparous and non-pregnant: Yes
- Age at study initiation: Young adult animals (male animals approx. 8 weeks, female animals approx. 12 weeks)
- Weight at study initiation: Animals of comparable weight (± 20% of the mean weight female: 211.8 g; male: 241.8 g)
- Housing: Single housing
- Diet: ad libitum
- Water: Tap water ad libitum
- Acclimation period: Acclimatization period of at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22°C +/- 3°C
- Humidity: 30 – 70%
- Air changes (per hr): Approx. 10
- Photoperiod (hrs dark / hrs light): 12 h / 12 h (6.00 a.m. – 6.00 p.m. / 6.00 p.m. – 6.00 a.m.)

Type of coverage:

semiocclusive

Vehicle:

corn oil

Details on dermal exposure:

TEST SITE
- Area of exposure: dorsal and dorsolateral parts of the trunk
- % coverage: About 40 cm² (corresponds to at least 10% of the body surface)

REMOVAL OF TEST SUBSTANCE
- Washing: Yes, with warm water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount applied: 3.33 g/kg bw
- Concentration: 60g /100 mL
- Constant volume or concentration used: yes
- For solids, paste formed: yes

Duration of exposure:

24 hours

Doses:

2000 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: At least once during each workday thereafter and individual body weights shortly before application (day 0), weekly thereafter and on the last day of observation.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, scoring of skin findings, mortality

Analytical examinations:

Stability of the test item preparation: Was determined indirectly by concentration control analysis.
Homogeneity of the test item preparation, Concentration control analysis of the test item preparation, Analysis of feed, Analysis of drinking water, Bedding and enrichment analysis

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:

No systemic clinical signs were observed during clinical examination.
Local effects: Yellowish discoloration of the application area was seen in all animals from study day 1 until study day 2.

Body weight:

The body weight of the male animals increased within the normal range throughout the study period. The body weight of the female animals increased in two animals within the normal range throughout the study period. Three female animals showed stagnation of body weight during the first week, while these animals gained weight in a normal range during the second week. As stagnation of body weight is generally known to occur as a consequence of the dermal application procedure, the body weight stagnation observed is considered to be unspecific.

Gross pathology:

No macroscopic pathologic abnormalities were noted in the animals (5 males and 5 females) examined on the last day of observation.

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

GLP and guideline compliant study

Additional information

Acute oral toxicity

Key study

In an acute oral toxicity study performed according to GLP and the OECD 423 Acute Toxic Class method, 2000 mg/kg of the test substance (preparations in corn oil Ph.Eur.) were administered by gavage to two test groups of three fasted Wistar rats each (6 females). The animals were observed for 14 days.

No mortality occurred and no clinical signs were observed.

The body weight of the animals increased within the normal range throughout the study period with one exception. In one animal the body weight increased normally during the first week, but this animal only slightly gained weight during the second week. This effect is observed at times in the rat strain used, because in the required age range the female animals have already reached the phase of slow growth.

There were no macroscopic pathological findings at the end of the observation period.

The acute oral LD50 was determined to be > 2000 mg/kg bw.

Acute dermal toxicity

Key study

In a GLP and OECD 402 guideline compliant acute dermal toxicity study (Limit Test), young adult Wistar rats (5 males and 5 females) were dermally exposed to a single dose of 2000 mg/kg bw of the test substance (as suspension in corn oil Ph.Eur.). The clipped application site (dorsal and dorso-lateral parts of the trunk, comprising at least 10% of the total body surface), was covered by semi-occlusive dressing during the 24-hour exposure period. The animals were observed for 14 days. In the study no mortality occurred and no clinical signs were observed.

The following test item-related local effects were recorded on the first two days after application: Yellowish discoloration of the application area in all animals.

The body weight of the male animals increased within the normal range throughout the study period. The body weight of the female animals increased in two animals within the normal range throughout the study period. Three female animals showed stagnation of body weight during the first week, while these animals gained weight in a normal range during the second week. As stagnation of body weight is generally known to occur as a consequence of the dermal application procedure, the body weight stagnation observed is considered to be unspecific.  

No macroscopic pathologic abnormalities were noted in the animals examined at the end of the study.

Accordingly, the acute dermal median lethal dose (LD50) was determined to be > 2000 mg/kg bw.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

Acute oral toxicity

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008 (CLP). The LD50 for acute oral toxicity was greater 2000 mg/kg bw. As a result the test substance is not considered to be classified for acute oral toxicity under Regulation (EC) No 1272/2008, as amended for the tenth time in Regulation (EU) No 2017/776.

 

Acute dermal toxicity

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008 (CLP). The LD50 for acute dermal toxicity was greater 2000 mg/kg bw. As a result the test substance is not considered to be classified for acute dermal toxicity under Regulation (EC) No 1272/2008, as amended for the tenth time in Regulation (EU) No 2017/776.