Strontium 3-hydroxy-4-[(1-sulfonato-2-naphthyl)azo]-2- naphthoate 5/2 hydrate

Endpoint summary

Description of key information

The oral LD50 value of R507-2 in Wistar rats was established to exceed 2000 mg/kg body weight.

The dermal LD50 value of R507-2 in Wistar rats was established to exceed 2000 mg/kg body weight.

The inhalation LC50 value of R507-2 in Wistar rats was found to exceed the maximum attainable concentration of 3 mg/L.

However based on the maximum concentration attainable in air, the minor temporary decrease in body weight, the absence of mortality, clinical symptoms and macroscopic findings, R507-2 does not have to be classified and has no obligatory labelling requirement for inhalation toxicity according to the GHS of Classification and labelling of Chemicals of the United nations (2004) and EC criteria for classification and labelling (1272/2008).

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:

7th November 2007- 23rd November 2007

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)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfield, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 9 weeks old
- Weight at study initiation: Does not exceed the mean.
- Fasting period before study: 20 hours.
- Housing: Animals were housed in 3 cages in labelled Macrolon cages containing sterilized sawdust as bedding material and paper as cage- enrichment.
- Diet (e.g. ad libitum): Free access to pelleted rodent diet.
- Water (e.g. ad libitum): Free access to tap water.
- Acclimation period: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 23.1 °C
- Humidity (%): 40-80 %
- Air changes (per hr): 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial light and 12 hours darkness per day.

IN-LIFE DATES: From: 07 November 2007 To: 23rd November 2007

Route of administration:

oral: gavage

Vehicle:

propylene glycol

Details on oral exposure:

VEHICLE
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed at NOTOX and on test substance data supplied by the sponsor.

Doses:

2000 mg/kg (10mL/kg) body weight

No. of animals per sex per dose:

3 animals per dose.

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Body weights were measured on Day 1, 8 and 15. Clinical observations were recorded daily.
- Necropsy of survivors performed: yes

Statistics:

No statistical analysis was performed.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No Mortality occurred.

Clinical signs:

other: Hunched posture and/or uncoordinated movements were noted in all animals on Day 1. In the first set of animals, red staining of the back was observed on Day 1 and red faeces were observed between Days 2 and 4. These findings were considered to be related

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Interpretation of results:

GHS criteria not met

Conclusions:

The oral LD50 value of R507-2 in Wistar rats was established to exceed 2000 mg/kg body weight.

According to the OECD 423 test guideline, the LD50 cut-off value was considered to exceed 5000 mg/kg body weight.

Based on these results, R507-2 does not have to be classified and has no obligatory labelling requirement for oral toxicity according to the GHS of Classification and labelling of Chemicals of the United nations (2004) and EC criteria for classification and labelling (1272/2008)

Executive summary:

R507 -2 was administered by oral gavage to two subsequent groups of three female Wistar rats at 2000 mg/kg body weight. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15)

No Mortality occurred.

Hunched posture and/or uncoordinated movements were noted in all animals on Day 1. In the first set of animals, red staining of the back was observed on Day 1 and red faeces were observed between Days 2 and 4. These findings were considered to be related to staining properties of the test substance and to be of no toxicological significance.

The body weight gain shown by animals over the study period was considered to be normal.

he oral LD50 value of R507-2 in Wistar rats was established to exceed 2000mg/kg body weight.

According to the OECD 423 test guideline, the LD50 cut-off value was considered to exceed 5000 mg/kg body weight.

Based on these results, R507-2 does not have to be classified and has no obligatory labelling requirement for oral toxicity according to the GHS of Classification and labelling of Chemicals of the United nations (2004) and EC criteria for classification and labelling (1272/2008).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

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:

27th March 2008 - 10 April 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.2 (Acute Toxicity (Inhalation))

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed concentration procedure

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 Deutschland, Sulzfield, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 15 weeks old
- Weight at study initiation: Does not exceed the mean.
- Fasting period before study: Not specified.
- Housing: Animals were housed in 5 animals per sex per cage in labelled Macrolon cages containing sterilized sawdust as bedding material and paper as cage- enrichment.
- Diet (e.g. ad libitum): Free access to pelleted rodent diet.
- Water (e.g. ad libitum): Free access to tap water.
- Acclimation period: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.9 - 21.3 °C
- Humidity (%): 40-80 %
- Air changes (per hr): 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial light and 12 hours darkness per day.

IN-LIFE DATES: From: 27 March 2008 To: 10 April 2008

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

oxygen

Mass median aerodynamic diameter (MMAD):

> 1 - <= 1.2 µm

Geometric standard deviation (GSD):

> 2.4 - < 3.3

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Exposure chamber
- Method of holding animals in test chamber: Animals were restrained in polycarbonate restraining tubes.
- Source and rate of air: 2.0 L/min
- System of generating particulates/aerosols: Aerosol was generated by administering the test substance to a stream of pressurized air by means of a brush feeder and an air mover. The aerosol was directly led through the exposure chamber. From the exposure chamber the test atmosphere was passed through a filter before it was released to the exhaust of the fume hood.
- Method of particle size determination: Particle size distribution was characterized twice during the exposure period. The samples were drawn (2 L/min) from the test atmosphere through a tube mounted in one the free animal ports of the middles sections of the exposure chamber. Samples were than collected with an 8 stage Marple personal cascade impactor. Amounts of test substance collected were gravimetrically. MMAD and GSD were determined.
- Temperature, humidity, pressure in air chamber: The temperature and relative humidity were measured with a humidity and temperature indicator and were recorded after the animals were placed in the experimental set up and at 30 minute intervals after initiation of the exposure. The probe was inserted in a tube mounted in one of the free animal ports of the middles section of the exposure chamber.

TEST ATMOSPHERE
- Brief description of analytical method used: Sample volumes were measured by means of a dry gas meter.
- Samples taken from breathing zone: yes

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

3.0 mg/L

No. of animals per sex per dose:

5 animals per sex per dose.

Control animals:

not specified

Details on study design:

Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Body weights wer measured on Day 1, 8 and 15. Clinical observations were recorded twice at 1 and 3 hours after exposure, on the day of the dosing and daily thereafter until day 15.
- Necropsy of survivors performed: yes

Statistics:

No statistical analysis was performed.

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 3 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Mortality:

No mortality occurred.

Clinical signs:

other: No signs of systemic toxicity were noted. Red staining of the fur due to deposition of test substance on the snout of the animals during exposure was noted in all animals throughout the entire observation period. Red faeces were noted in all animals from

Body weight:

Slight body weight loss was shown by the males between Day 1 and 8. This body weight loss was considered minor based on the absence of any corroborative findings in these animals. The mean body weight gain shown by females and males over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.

Gross pathology:

Macroscopic post mortem examination of the animals at termination revealed dark red discolouration of the caecum in one male and two females and reddish discolouration of the tail in all animals.

An average exposure concentration of 3 mg/L was attained instead of the target concentration of 5 mg/L. During method development in the first instance a spiral feeder was used to administer test substance to the air mover. Although different spirals were used no stable delivery of test substance to the air mover was attained, due to clogging of the spiral by the test substance. The spiral feeder was replaced by a brush feeder for the delivery of the test substance. A more stable concentration was attained and no clogging of the apparatus occurred. It was concluded that the brush feeder was the most suitable feeding apparatus.

In view of the effort spent to generate a test atmosphere with R507 -2 and the generation efficiency during exposure to animals, which was relatively high, 3 mg/L was considered the maximum attainable concentration which could be generated in air for 4 hours.

Interpretation of results:

GHS criteria not met

Conclusions:

The LC50 value of R507-2 in Wistar rats was found to exceed the maximum attainable concentration of 3 mg/L.

However based on the maximum concentration attainable in air, the minor temporary decrease in body weight, the absence of mortality, clinical symptoms and macroscopic findings, R507-2 does not have to be classified and has no obligatory labelling requirement for inhalation toxicity according to the GHS of Classification and labelling of Chemicals of the United nations (2004) and EC criteria for classification and labelling (1272/2008).

Executive summary:

R507-2 was administered by inhalation for 4 hours to one group of five males and five female Wistar rats. Animals were subjected to daily observations and weekly determinations of body weight. Macroscopic examination was performed after terminal sacrifice (day 15)

The mean actual concentration was 3 +/-0.7 mg/L. The concentration was considered the maximum attainable concentration of R507 -2 which could be generated at a stable level for 4 hours. The nominal concentration was 14.9 mg/L. The generation efficiency was 20 %.

MMAD and GSD were determined twice. The MMAD was 1.2 micrometeres and 1.0 micrometres respectively and the GSD was 2.4 and 3.3 respectively.

No mortality occurred.

No signs of systemic toxicity were noted.

Red staining of the fur due to deposition of test substance on the snout of the animals during exposure was noted in all animals throughout the entire observation period. Red faeces were noted in all animals from Day 2 to day 5. This finding is considered to be due to grooming of the fur and the subsequent ingestion of the test substance.

The mean body weight gain shown by females and males over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.

Macroscopic post mortem examination of the animals at termination revealed dark red discolouration of the caecum in one male and two females and reddish discolouration of the tail in all animals.

The LC50 value of R507 -2 in Wistar rats was found to exceed the maximum attainable concentration of 3 mg/L.

However based on the maximum concentration attainable in air, the minor temporary decrease in body weight, the absence of mortality, clinical symptoms and macroscopic findings, R507-2 does not have to be classified and has no obligatory labelling requirement for inhalation toxicity according to the GHS of Classification and labelling of Chemicals of the United nations (2004) and EC criteria for classification and labelling (1272/2008).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

3 mg/m³ air

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:

13th February 2008- 27th February 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

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: Charles River Deutschland, Sulzfield, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 9 weeks old
- Weight at study initiation: Does not exceed the mean.
- Fasting period before study: Not specified.
- Housing: Animals were housed individually in labelled Macrolon cages containing sterilized sawdust as bedding material and paper as cage- enrichment.
- Diet (e.g. ad libitum): Free access to pelleted rodent diet.
- Water (e.g. ad libitum): Free access to tap water.
- Acclimation period: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.7 - 21.6 °C
- Humidity (%): 39 - 63 %
- Air changes (per hr): 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial light and 12 hours darkness per day.

IN-LIFE DATES: From: 13 February 2008 To: 27 February 2008

Type of coverage:

occlusive

Vehicle:

propylene glycol

Details on dermal exposure:

TEST SITE
- Area of exposure: Back
- % coverage: 10 %
- Type of wrap if used: Covered with aluminium foil and Coban elastic bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Residual test substance removed with tap water.
- Time after start of exposure: 24 hours.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg (10 mL/kg)

Duration of exposure:

24 Hours

Doses:

Single dose

No. of animals per sex per dose:

5 animals per sex per dose.

Control animals:

not specified

Details on study design:

Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Body weights were measured on Day 1, 8 and 15. Clinical observations were recorded daily.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other

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:

other: Hunched posture, piloerection and/or chromodacryorrhoea were noted among both males and females. In addition, lethargy, flat posture, uncoordinated movements, quick breathing, shallow respiration, ptosis and/or hypothemia were noted in all males. The anim

Gross pathology:

At macroscopic post mortem examination, isolated grey/white foci, enlargement and dark red discoloration were found in the papillary process in the liver of one male. Macroscopic post mortem examination of the other animals at termination did not reveal any abnormalities.

Interpretation of results:

GHS criteria not met

Conclusions:

The dermal LD50 value of R507-2 in Wistar rats was established to exceed 2000 mg/kg body weight.

Based on these results, R507-2 does not have to be classified and has no obligatory labelling requirement for dermal toxicity according to the GHS of Classification and labelling of Chemicals of the United nations (2004) and EC criteria for classification and labelling (1272/2008)

Executive summary:

R507 -2 was administered by a single dermal application to two subsequent groups of 5 female Wistar rats and 5 male rats at 2000 mg/kg body weight for 24 hours. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15)

No Mortality occurred.

Hunched posture, piloerection and/or chromodacryorrhoea were noted among both males and females. In addition, lethargy, flat posture, uncoordinated movements, quick breathing, shallow respiration, ptosis and/or hypothemia were noted in all males. The animals had recovered from the symptoms between Days 2 and 3.

These symptoms were occasionally seen in dermal toxicity studies. Based on their mild nature and short duration, these symptoms were considered to be of no toxicological significance.

Scales and/or scabs were seen in the treated skin area of the majority of females during the observation period.

Red staining of the treated skin area and/or other body parts of the animals was noted during the observation period and was considered to be related to staining properties of the test substance.

The changes noted in the body weight gain in males and females were within the range expected for rats used in this type of study.

At macroscopic post mortem examination, isolated grey/white foci, enlargement and dark red discoloration were found in the papillary process in the liver of one male. Macroscopic post mortem examination of the other animals at termination did not reveal any abnormalities.

The dermal LD50 value of R507-2 in Wistar rats was established to exceed 2000 mg/kg body weight.

Based on these results, R507-2 does not have to be classified and has no obligatory labelling requirement for dermal toxicity according to the GHS of Classification and labelling of Chemicals of the United nations (2004) and EC criteria for classification and labelling (1272/2008)

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Justification for classification or non-classification

According to EC criteria for classification and labelling (1272/2008) the test substance R507 -2 is not classified for acute toxicity.