Reaction mass of tetrasodium 7,7'-(carbonyldiimino)bis[4-hydroxy-3-[(2-methyl-4-sulphonatophenyl)azo]naphthalene-2-sulphonate] and tetrasodium 4-[[1-hydroxy-6-[[[[5-hydroxy-6-[(2-methyl-4-sulphonatophenyl)azo]-7-sulphonato-2-naphthyl]amino]carbonyl]amino]-3-sulphonato-2-naphthyl]azo]benzoate and tetrasodium 4,4'-[carbonylbis[imino(1-hydroxy-3-sulphonatonaphthalene-6,2-diyl)azo]]dibenzoate

Endpoint summary

Description of key information

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant, guideline study, available as unpublished report, no restrictions, fully adequate for assessment

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Qualifier:

according to guideline

Guideline:

other: Japan MAFF Testing Guideline of 12 Nosan No. 8147

GLP compliance:

yes (incl. QA statement)

Remarks:

Bioassay, Labor für biologische Analytik GmbH, Im Neuenheimer Feld 515-519, 69120 Heidelberg

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga GmbH, Germany
- Age at study initiation: Young adult animals (approx. 10 weeks)
- Weight at study initiation: mean weight of 6 animals: 177.7 g
- Fasting period before study: 16 hours before administration
- Housing: Makrolon cage, type III, single housing with Bedding: H 15005-29; Ssniff, Spezialitäten GmbH (Experimental Animal Diets Inc., 59494 Soest, Germany) and Enrichment: Wooden gnawing blocks (Type NGM E-022) ; ABEDD® LAB & VET Service GmbH, Hasnerstraße 84/6; 1160 Wien – Austria
- Diet: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany, ad libitum
- Water: Tap water, ad libitum
- Acclimation period: at least 5 days before

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): approx. 10
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Remarks:

0.5% solution in deionized water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 20 g/mL
- Amount of vehicle (if gavage): 10 mL/kg bw
- Justification for choice of vehicle: A good homogeneity in water could not be guaranteed, because the test item preparation was a suspension. Therefore a 0.5% solution of CMC in deionized water was applied

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

6

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 d
- Frequency of observations and weighing: Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation. Clinical signs for each animal were recorded several times on the day of administration and at least once during each workday thereafter. A check for any dead or moribund animals was made at least once each workday.
- 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. No histological examinations were performed.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Mortality:

No mortality occurred.

Clinical signs:

other: All animals of the first 2000 mg/kg bw. test group showed a red discoloration of feces from study day 1 until day 3 and red discolored urine from day 2 until day 3. In addition, two of these animals showed impaired general state and piloerection from hour

Gross pathology:

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

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

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:

other: GLP compliant, guideline study, available as unpublished report, no restrictions, fully adequate for assessment

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

other: Japan MAFF Testing Guideline of 12 Nosan No. 8147

GLP compliance:

yes (incl. QA statement)

Remarks:

Bioassay Labor für biologische Analytik GmbH, 69120 Heidelberg, Germany

Test type:

standard acute method

Limit test:

yes

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
- Age at study initiation: male animals approx. 8 weeks, female animals approx. 12 weeks
- Mean weight at study initiation (mean weight ± SD): males: 227.6 ± 2.88 and females 206.8 ± 2.17 g
- Housing: Single housing, Makrolon cage, type III with bedding (H 15005-29; Ssniff, Spezialitäten GmbH (Experimental Animal Diets Inc., 59494 Soest, Germany)) and enrichment (Wooden gnawing blocks (Type NGM E-022) ; ABEDD® LAB & VET Service GmbH, Hasnerstraße 84/6; 1160 Wien – Austria)
- Diet: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany, ad libitum
- Water: Tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30-70
- Air changes (per hr): Approx. 10
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Vehicle:

CMC (carboxymethyl cellulose)

Remarks:

0.5% solution in deionized water

Details on dermal exposure:

TEST SITE
- Area of exposure: Single application to the clipped epidermis (dorsal and dorsolateral parts of the trunk).
- % coverage: approx. 10 (about 40 cm²)
- Type of wrap if used: semi-occlusive dressing.

REMOVAL OF TEST SUBSTANCE
- Washing: rinsing of the application site with warm water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount applied: 5.71 mL/kg bw
- Concentration: 35g/100 mL
- Form of application: suspension
- Preparation test item: The test item preparation was produced for the 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.

VEHICLE
A good homogeneity in water could not be guaranteed, because the test item preparation was a suspension. Therefore a 0.5% solution of CMC in deionized water was applied.

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: Individual body weights shortly before application (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.
- Scoring of skin findings: Individual readings 30 – 60 minutes after removal of the semi-occlusive dressing (day 1), several times and on the last day of observation
- Mortality: A check for any dead or moribund animals was made at least once each workday.
- 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.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

other: No systemic clinical signs were observed.

Gross pathology:

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

Other findings:

Local effects:
- Males: Due to the reddish discoloration of the application area, no erythema was assessable on study day 1 only. Moderate erythema (grade 3) was seen in four animals from day 2 until day 3, followed by well-defined erythema from day 6 until day 9 and very slight erythema (grade 1) on day 10. The fifth animal showed also moderate erythema from day 2 until day 3, but followed by very slight erythema on day 6, 7 and 10. Four animals showed severe edema (grade 4) on day 1, followed by moderate edema (grade 3) from day 2 until day 3 and very slight edema (grade 1) on day 6 and 7. The fifth male animal showed moderate edema from day 1 until day 3, followed by slight edema (grade 2) on day 6 and 7 and very slight edema on day 8 and 9. Red test item residues and a reddish discoloration of the application area could be noted from day 2 until day 9 in all animals.
- Females: Due to the reddish discoloration of the application area, no erythema was assessable on day 1 only. Moderate erythema (grade 3) was seen in four animals from day 2 until day 3, followed by well-defined erythema (grade 2) from day 6 until day 9 or day 10. One of these animals showed very slight erythema (grade 1) on day 10. In the fifth animal showed well-defined erythema (grade 2) was observed from day 2 until day 10. Four animals showed moderate edema (grade 3) from day 1 until day 2, ensued by slight edema (grade 2) on day 3. On day six very slight edema (grade 1) could be noted in three of these animals, while the fourth animal still showed slight edema on day 6. In the fifth animal moderate edema was observed on day 1, followed by slight edema from day 2 until day 3 and very slight edema on day 6. Red test item residues and a reddish discoloration of the application area could be noted from day 2 until day 10 in all animals.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Additional information

Acute toxicity: oral

An acute oral toxicity study was conducted according to OECD 423 and in compliance with GLP. Six female Wistar rats were exposed to 2000 mg/kg bw test substance dissolved in 0.5% CMC-solution by oral gavage. No mortality was observed. Impaired general state and piloerection were observed in 4 animals. Red discoloration of feces was observed in all animals and discoloration of urine in 3 animals. No adverse effects on body weight were observed. There were no macroscopic pathological findings in any of the animals. Under the conditions of the test the LD50 was determined to be greater than 2000 mg/kg bw.

Three albino rats per sex per dose were exposed via oral gavage. Animals received 2000 mg/kg bw of the test substance (containing 20% of the active ingredient). At the end of the study period animals were necropsied. No mortality was observed. Piloerection, hunched posture, and dyspnea were seen. The animals recovered within 6 to 8 days. Gross pathology showed no deviations. The LD50 was determined to be >2000 mg/kg bw.

 

Acute toxicity: dermal 

An acute dermal toxicity study was performed according to OECD 402 and in compliance with GLP. Five Wistar rats per sex were exposed to 2000 mg/kg bw as suspension in 0.5 % CMC-solution. The test substance was applied to clipped application site (dorsal and dorso-lateral parts of the trunk, comprising at least 10% of the total body surface) and was covered by semi-occlusive dressing during the 24-hour exposure period. The animals were observed for 14 days. Neither mortality nor signs of systemic toxicity were observed in the animals. Local effects occurred within the first 10 days after application. Very slight to moderate erythema (grade 1 to 3) and very slight to severe edema (grade 1 to 4) were observed. Erythema was not assessable due to reddish discoloration of the application area (day 1). Some stagnation of body weight gain was observed but was considered to be unspecific. No macroscopic pathologic abnormalities were noted in all animals examined at the end of the study. Under the conditions of the test the LD50 was determined to be greater than 2000 mg/kg bw.

Justification for selection of acute toxicity – oral endpoint
There are two studies available. The study performed with the highest concentration active ingredient (90%) was selected as key.

Justification for selection of acute toxicity – dermal endpoint
There is only one study available.

Justification for classification or non-classification

As the oral LD50 and the dermal LD50 were determined to be >2000 mg/kg bw, classification for oral and dermal acute toxicity is not warranted in accordance with EU Classification, Labeling and Packaging of Substances and Mixtures (CLP) Regulation No. 1272/2008.

 

Due to lacking data classification for acute inhalation toxicity is not possible in accordance with EU Classification, Labeling and Packaging of Substances and Mixtures (CLP) Regulation No. 1272/2008.