Benzoic acid, 4-[2-(2-hydroxy-6-sulfo-1-naphthalenyl)diazenyl]-, strontium salt (2:1)

Endpoint summary

Description of key information

Acute oral toxicity: LD50 > 2000 mg/kg bw; Limit-Test, GLP, OECD Guideline Study, Safepharm Laboratories Ltd, 1235/003, 1998. 
The test substance is virtually nontoxic after a single oral administration. 
Acute dermal toxicity: LD50 > 2000 mg/kg bw; Limit-Test, GLP, OECD Guideline Study, Safepharm Laboratories Ltd, 1235/054, 1998
The test substance is virtually nontoxic after a single dermal application.

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:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

adopted 24 February 1987

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

other: Sprague-Dawley (Crl : CD ® (SD) IGS BR)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Ltd ., Margate, Kent, U.K.
- Age at study initiation: eight to twelve weeks.
- Weight at study initiation: males: 201 - 217g, females: 200 - 214g,
- Fasting period before study: overnight fast immediately before dosing and for approximately three to four hours after dosing
- Housing: up to five by sex in solid floor polypropylene cages furnished with woodflakes
- Diet: Rat and Mouse Expanded Diet No . 1, Special Diet Services Limited, Witham, Essex, U.K.), ad libitum
- Water: mains drinking water, ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 23
- Humidity (%): 54 - 67
- Air changes (per hr): approximately 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 / 12

Route of administration:

oral: gavage

Vehicle:

other: distilled water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg / ml
- Amount of vehicle (if gavage): The volume administered to each animal was calculated according to its fasted bodyweight at the time of dosing.
- Justification for choice of vehicle: No data

DOSE VOLUME APPLIED: 10 ml / kg

DOSAGE PREPARATION (if unusual):
- For the purpose of the study the test material was freshly prepared, as required, as a solution at the appropriate concentration in distilled water.
- The concentration, homogeneity and stability of the test material preparations were not determined by analysis.

Doses:

2000 mg/kg bodyweight

No. of animals per sex per dose:

5

Control animals:

not specified

Details on study design:

• Range-finding Study:
2000 mg / kg bodyweight were administered at a concentration of about 200 mg / mL at a dose volume of 10 mL / kg to one male and one female rat, respectively.
- Duration of observation period following administration: five days.
- Frequency of observations and weighing: The animals were observed for deaths or overt signs of toxicity ½, 1 2, and 4 hours after dosing and subsequently once daily for five days. Individual bodyweights were recorded on the day of dosing to allow calculation of individual treatment volumes.
- Necropsy of survivors performed: no
There were no deaths or clinical signs of toxicity.

• Main Study:
Based on the information of the range-finding study, a dose level of 2000 mg / kg bodyweight was selected for the main study.
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed for deaths or overt signs of toxicity ½, 1, 2 and 4 hours after dosing and subsequently once daily for 14 days. Individual bodyweights were recorded prior to dosing on Day 0 and on Days 7 and 14.
- Necropsy of survivors performed: yes, at the end of the study the animals were killed by cervical dislocation and subjected to gross pathological examination. This consisted of an external examination and opening of the abdominal and thoracic cavities for examination of major organs. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality. No signs of systemic toxicity.

Mortality:

There were no deaths.

Clinical signs:

other: No signs of systemic toxicity were noted during the study. The faeces of all animals was stained orange one and two days after treatment. The urine of all females was also stained orange during this period.

Gross pathology:

No abnormalities were noted at necropsy.

Other findings:

No other findings.

Table 1: Mean body weight
		Mean	SD
Day 0	Males	208.6	5.86
	Females	206.0	5.83
Day 7	Males	269.2	9.68
	Females	238.0	8.80
Day 14	Males	315.4	17.52
	Females	257.2	11.82

Table 2: Body weight gain (g) during week
		Mean	SD
Week 1	Males	60.6	8.11
	Females	32.0	3.94
Week 2	Males	46.2	8.93
	Females	19.2	3.90

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

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 - Guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

adopted 24 February 1987

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

other: Sprague-Dawley (Crl : CD ® (SD) IGS BR)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Ltd ., Margate, Kent, UK
- Age at study initiation: eight to twelve weeks.
- Weight at study initiation: males: 205 - 218 g, females: 200 - 221 g,
- Fasting period before study: overnight fast immediately before dosing and for approximately three to four hours after dosing
- Housing: The animals were housed in suspended polypropylene cages furnished with woodflakes. The animals were housed individually during the 24-hour exposure period and in groups of five, by sex, for the remainder of the study.
- Diet: Rat and Mouse Expanded Diet No . 1, Special Diet Services Limited, Witham, Essex, UK), ad libitum
- Water: mains drinking water, ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 21
- Humidity (%): 51 - 65
- Air changes (per hr): approximately 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 / 12

Type of coverage:

semiocclusive

Vehicle:

other: The test material was moistened with distilled water.

Details on dermal exposure:

TEST SITE
- On the day before treatment the back and flanks of each animal were clipped free of hair using veterinary dippers.
- Area of exposure: back and flanks
- % coverage: approximating to 10% of the total body surface area
- Type of wrap if used: A piece of surgical gauze was placed over the treatment area and semioccluded with a piece of self-adhesive bandage. The animals were caged individually for the 24-hour exposure period. Shortly after dosing the dressings were examined to ensure that they were securely in place.

REMOVAL OF TEST SUBSTANCE
- Washing: the bandage was carefully removed and the treated skin and surrounding hair wiped with cotton wool moistened with
distilled water to remove any residual test material.
- Time after start of exposure: 24h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg / kg bw
- Concentration (if solution): undiluted
- Constant volume or concentration used: yes

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: once daily
- Frequency of weighing: prior to application of the test material on day 0 and on day 7 and 14.
- Necropsy of survivors performed: yes, animals were killed by cervical dislocation and subjected to gross pathological examination. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.
- Other examinations performed: clinical signs, body weight, test sites were examined for evidence of primary irritation and scored according to Draize J.H. (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy 01 Sciences, Washington DC p. 31.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality. No signs of systemic toxicity.

Mortality:

There were no deaths.

Clinical signs:

other: No signs of systemic toxicity were noted during the study. Orange-coloured staining caused by the test material was noted at the treated skin sites of all animals and prevented an evaluation of the degree of erythema on Day 1. No signs of skin irritation

Gross pathology:

No abnormalities were noted at necropsy.

Other findings:

Signs of toxicity (local):
On day 1, scoring for erythema due to orange staining was not possible. No signs of dermal irritation were detected until the end of the study.

Table 1: Mean body weight
		Mean	SD*
Day 0	Males	213.4	5.22
	Females	208.6	8.62
Day 7	Males	256.6	18.37
	Females	221.6	9.29
Day 14	Males	311.4	23.16
	Females	238	11.98

Table 2: Body weight gain (g) during week 1 and 2
		Mean	SD*
Week 1	Males	43.2	15.91
	Females	13.0	3.61
Week 2	Males	54.8	14.81
	Females	16.4	3.51

* standard deviation

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Additional information

There are valid in vivo data available for the assessment of the acute oral and dermal toxicity of the test item.

Acute Oral Toxicity

In the key study, performed as a limit test according to GLP and OECD Guideline 401 (Acute Oral Toxicity), fasted, 8 to 12 weeks old, Sprague Dawley rats (Crl : CD ® (SD) IGS BR, 5/sex/dose) were given a single oral dose (gavage) of the test item (analytical purity unknown) at a single dose of 2000 mg/kg bw (Safepharm Laboratories Ltd, 1235/003, 1989). The test item was diluted in distilled water at a concentration of 0.2 mg/mL and administered at a volume dosage of 10 ml/kg. The animals were observed subsequently for a period of 14 days. All animals were killed and subjected to gross pathology. No mortality occurred during the observation period. No signs of systemic toxicity were noted during the study. The faeces of all animals and the urine of all females were stained orange one to two days after treatment. All animals showed an expected gain in bodyweight during the study. No abnormalities were noted at necropsy.

Acute Dermal Toxicity

In the key study,performed as a limit test according to GLP and OECD Guideline 402 (Acute Dermal Toxicity), 8 to 12 weeks old Sprague Dawley rats (Crl : CD ® (SD) IGS BR, 5/sex/dose) were given a single semi-occluded dermal application at a dose level of the test item (analytical purity unknown) of 2000 mg/kg bodyweight (Safepharm Laboratories Ltd, 1235/054, 1989). The exposure period was 24 h and the observation period 14 days. No mortality occurred during the observation period. There were no signs of systemic toxicity or skin irritation during the study. There were no treatment related necropsy findings or changes in body weight.

Justification for classification or non-classification

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

The available experimental test data are reliable and suitable for the purpose of classification under Regulation (EC) No.1272/2008. No mortality occurred in rats treated with the limit dose in OECD testing guideline studies. Based on the data, classification for acute oral or dermal toxicity is not warranted under Regulation (EC) No.1272/2008.