Chromate(5-), bis[4-(hydroxy-κO)-7-[2-(2-hydroxy-1-naphthalenyl)diazenyl]-3-[2-[2-(hydroxy-κO)- 3-nitro-5-sulfophenyl]diazenyl]-2-naphthalenesulfonato(4-)]-, sodium (1:5)

Endpoint summary

Description of key information

LD50 (oral) > 2000 mg/kg bw

LD50 (dermal) > 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:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
-Source: BRL Ltd., Basel, Switzerland
-Age at study initiation: Approx. 9 weeks.
-Weight at study initiation:
male: 206-231 g
female: 161 - 188 g
-Number of animals 5 ♂ and 5 ♀
-Housing: Group housing of 5 animals per sex per cage in labelled polycarbonate cages containing purified sawdust as bedding material
-Fasting period before study: Food was withheld overnight prior to dosing until approximately 3-4 hours after administration of the test substance.
-Diet: Free access to standard pelleted laboratory animal diet
-Water: Free access to tap-water
-Acclimation period: 5 days before start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
-Temperature (°C): 21 °C
-Humidity (%): 55%
-Air changes (per hr): 15 air changes per hour
-Photoperiod (hrs dark / hrs light): 12 hours cycle dark/light with artificial fluorescent light

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
-Amount of vehicle (if gavage): Dose volume 10 ml/kg body weight.

Doses:

2000 mg/kg b.w

No. of animals per sex per dose:

5 per sex per dose

Control animals:

no

Details on study design:

-Duration of observation period following administration: 15 days
-Frequency of observations Mortality/Viability: twice daily.
-Frequency of observations clinical signs: at periodic intervals on the day of dosing (day 1) and once daily thereafter. the time of onset, degree and duration were recorded.
-Frequency of observations and weighing: days 1 (pre-administration), 8 and 15
-Necropsy of survivors performed: yes
All animals surviving to the end of the observation period (day 15) were sacrificed by oxygen/carbon dioxide asphyxiation. All animals assigned to the study were subjected to necropsy and descriptions of all macroscopic abnormalities recorded.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study period.

Clinical signs:

other: No clinical signs of ill health or behavioural changes were observed during the study period. Blue/black discolouration of the tail and dark appearance of the faeces, noted in all animals, was ascribed to the staining properties of the test substance and

Gross pathology:

Macroscopic post mortem examination of the animals at termination revealed bluish discolouration of the gastro-intestinal wall and of the mesenteric lymph node and dark discolouration of the kidneys in all animals. In all males bluish discolouration of the testes was noted at postmortem.

Interpretation of results:

other: Not classified according to the CLP Regulation (EC 1272/2008)

Conclusions:

The oral LD50 value exceed 2000 mg/kg body weight.

Executive summary:

The oral acute toxicity of the substance was evaluated with a limit test to albino rat Wistar, according to the method B.1 of EEC-Directive 92/69 EEC. Two groups, each of 5 males and 5 females, were treated with a 2000 mg/kg bw single dose of the substance suspended in an aqueous solution and administered by oral gavage. Symptoms and mortality after administration were recorded during the observation period of 14 days. Thereafter all animals were submitted to necropsy and macroscopic examination.

No mortality occurred during the study period and no significant toxicological effects were observed. The oral LD50 value of the substance in rats was established to exceed 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

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:

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

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
-Source: BRL Ltd., Basel, Switzerland
-Age at study initiation: Approx. 9 weeks.
-Weight at study initiation:
male: 204-217 g
female: 167 - 179 g
-Number of animals 5 ♂ and 5 ♀
-Housing: Individually housed in labelled polycarbonate cages containing purified sawdust as bedding material
-Diet: Free access to standard pelleted laboratory animal diet
-Water: Free access to tap-water
-Acclimation period: 5 days before start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
-Temperature (°C): 21 °C
-Humidity (%): 55%
-Air changes (per hr): 15 air changes per hour
-Photoperiod (hrs dark / hrs light): 12 hours cycle dark/light with artificial fluorescent light

Type of coverage:

occlusive

Vehicle:

water

Remarks:

tap water

Details on dermal exposure:

TEST SITE
-Area of exposure: 5x7 cm on the back of the animal was clipped
-% coverage:
5 x 5 cm for male
3.5x5 cm for female
- Type of wrap if used: gauze patch fixed successively to aluminium foil and flexible bandage with drops of petrolatum.

TEST MATERIAL
-Volume: 10 ml/kg

Duration of exposure:

24 hours

Doses:

2000 mg/kg b.w

No. of animals per sex per dose:

5 per sex per dose

Control animals:

no

Details on study design:

-Duration of observation period following administration: 14 days
-Frequency of observations Mortality/Viability: Twice daily
-Frequency of observations clinical signs: At periodic intervals on the day of treatment
(day 1) and once daily thereafter.
-Frequency of observations and weighing: Days 1 (pre-administration), 8 and 15.
-Necropsy of survivors performed: yes
All animals surviving to the end of the observation period (day 15) were sacrificed by oxygen/carbon dioxide asphyxiation. All animals assigned to the study were subjected to necropsy and descriptions of all macroscopic abnormalities recorded.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study period.

Clinical signs:

other: No clinical signs of ill health or behavioural changes were observed during the study period. Black discolouration of the dorsal skin was ascribed to the staining properties of the test substance and considered not to represent a sign of toxicity.

Gross pathology:

Macroscopic post mortem examination of the animals at termination revealed pelvic dilation of the right kidney in one male only. Renal pelvic dilation is a common finding in animals of this age and strain and therefore considered not related to treatment.

Interpretation of results:

other: Not classified according to the CLP Regulation (EC 1272/2008)

Conclusions:

The dermal LD50 exceed 2000 mg/kg body weight.

Executive summary:

The dermal acute toxicity of the substance was evaluated with a limit test to albino rat Wistar, according to the method Directive 92/69/EEC, Annex V of the EEC directive 67/548/EEC; Part B.3 (1992). A 2000 mg/kg bw single dose of the substance was suspended in an aqueous solution and was administered to two groups each of 5 males and 5 females. A gauze patch fixed to aluminium foil and flexible bandage was applied for 24 h to the skin previously shaved. Clinical signs and mortality after administration were recorded during the observation period of 14 days after treatment. Thereafter all animals were submitted to necropsy and macroscopic examination.

No mortality occurred during the study period and no significant toxicological effects were observed. The dermal LD50 value of the substance in rats of both sexes over a period of 14 days was established to exceed 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Additional information

ACUTE ORAL TOXICITY

The oral acute toxicity of the substance was evaluated with a limit test to albino rat Wistar, according to the method B.1 of EEC-Directive 92/69 EEC. Two groups, each of 5 males and 5 females, were treated with a 2000 mg/kg bw single dose of the substance suspended in an aqueous solution and administered by oral gavage. Symptoms and mortality after administration were recorded during the observation period of 14 days. Thereafter all animals were submitted to necropsy and macroscopic examination.

No mortality occurred during the study period and no significant toxicological effects were observed. The oral LD50 value of the substance in rats was established to exceed 2000 mg/kg bw.

ACUTE DERMAL TOXICITY

The dermal acute toxicity of the substance was evaluated with a limit test to albino rat Wistar, according to the method B.3 of EEC-Directive 92/69 EEC. A 2000 mg/kg bw single dose of the substance was suspended in an aqueous solution and was administered to two groups each of 5 males and 5 females. A gauze patch fixed to aluminium foil and flexible bandage was applied for 24 h to the skin previously shaved. Clinical signs and mortality after administration were recorded during the observation period of 14 days after treatment. Thereafter all animals were submitted to necropsy and macroscopic examination.

No mortality occurred during the study period and no significant toxicological effects were observed. The dermal LD50 value of the substance in rats of both sexes over a period of 14 days was estimated to exceed 2000 mg/kg bw.

Justification for classification or non-classification

In the CLP Regulation (EC 1272/2008) acute toxicity is defined as “those adverse effects occurring following oral or dermal administration of a single dose of a substance or a mixture, or multiple doses given within 24 hours, or an inhalation exposure of 4 hours”. A substance can be allocated to one of four toxicity categories based on acute toxicity by the oral, dermal or inhalation route. The numeric criteria based on the acute toxicity estimates (ATE) are presented in Annex I: 3.1.2.1, Table 3.1.1.

Based on the results of acute oral and dermal toxicity studies performed on the test substance, no classification for acute toxicity is warranted under the CLP Regulation (EC 1272/2008).