Dihydrogen (ethyl)[4-[[4-[ethyl(3-sulphonatobenzyl)amino]phenyl](2-sulphonatophenyl)methylene]cyclohexa-2,5-dien-1-ylidene](3-sulphonatobenzyl)ammonium, aluminium salt

Endpoint summary

Description of key information

Acute oral toxicity: 

The acute oral toxicity dose (LD50) was considered based on different studies conducted on rats for the test chemical. The studies concluded that the LD50 value is >2000 mg/kg bw, for acute oral toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral toxicity.

Acute Inhalation Toxicity:

The acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to very low vapour pressure of the test chemical. Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver. 

 

Acute Dermal toxicity:

The acute dermal toxicity dose (LD50) was considered based on different studies conducted on rats for the test chemical. The studies concluded that LD50 value is >2000 mg/kg bw, for acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute dermal toxicity.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

experimental data from various test chemicals

Justification for type of information:

Data is summarized based on the available information from various read across test chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

WoE report is based on 3 acute oral toxicity studies as - WoE 2, WoE 3 and WoE 4.
Acute Oral toxicity test was carried out to study the effects of the test chemicals on rodents.

GLP compliance:

not specified

Test type:

other: not specified

Limit test:

no

Species:

rat

Strain:

other: 2. Wistar 3. not specified 4. not specified

Sex:

not specified

Details on test animals or test system and environmental conditions:

2. not specified
3. not specified
4. not specified

Route of administration:

oral: unspecified

Vehicle:

not specified

Details on oral exposure:

2. not specified
3. not specified
4. not specified

Doses:

2. 2000 mg/kg bw
3. 2000 mg/kg bw
4. 2000 mg/kg bw

No. of animals per sex per dose:

2. not specified
3. not specified
4. not specified

Control animals:

not specified

Details on study design:

2. not specified
3. not specified
4. not specified

Statistics:

2. not specified
3. not specified
4. not specified

Preliminary study:

2. not specified
3. not specified
4. not specified

Sex:

not specified

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

2. No mortality was observed at 2000 mg/kg bw in treated animals.
3. No mortality was observed at 2000 mg/kg bw.
4. No mortality was observed in treated animals.

Clinical signs:

other: 2. not specified 3. not specified 4. not specified

Gross pathology:

2. not specified
3. not specified
4. not specified

Other findings:

2. not specified
3. not specified
4. not specified

Interpretation of results:

other: Not classified

Conclusions:

According to CLP regulation, the given test chemical cannot be classified for acute oral toxicity, as the LD50 value is >2000 mg/kg bw.

Executive summary:

In different studies, the given test chemical has been investigated for acute oral toxicity to a greater or lesser extent. Often are the studies based on in-vivo experiments in rodents, i.e. most commonly in rats for test chemical. The studies are summarized as below –

 

The reported study was mentioned in peer-reviewed journal and conducted to determine acute oral toxicity dose by using the given test chemical in adult Wistar rat at the dose concentration of 2000 mg/kg bw orally. Animals were observed for mortality. No mortality was observed at 2000 mg/kg bw. Hence, the Lethal concentration value (LD50) was considered to be >2000 mg/kg bw, when adult Wistar rat was treated with the given test chemical orally.

 

The above study is supported with another study mentioned in handbook and authoritative database for the given test chemical. The acute oral toxicity study was performed in rat at the dose concentration of 2000 mg/kg bw. Animals were observed for mortality. No mortality was observed at 2000 mg/kg bw. Hence, the Lethal concentration value (LD50) was considered to be >2000 mg/kg bw, when rat was treated with the given test chemical orally.

 

Both the above studies are further supported with the data available in handbook and authoritative database using the given test chemical in rat at the dose concentration of 2000 mg/kg bw. The animals were observed for mortality. No mortality was observed at 2000 mg/kg bw in treated animals. Hence, the Lethal concentration value (LD50) was considered to be >2000 mg/kg bw, when rat was treated with the given test chemical orally.

 

Thus, based on the above summarised studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral toxicity.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Data is Klimisch 2 and from publication.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

other:

Endpoint conclusion

Quality of whole database:

Waiver

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

experimental data from various test chemicals

Justification for type of information:

Data is summarized based on the available information from various read across test chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

WoE report is based on 2 acute dermal toxicity studies as- WoE 2 and WoE 3.
Acute dermal toxicity test was carried out to study the effects of the test chemicals on rodents.

GLP compliance:

not specified

Test type:

other: not specified

Limit test:

no

Species:

rat

Strain:

other: 2. Sprague-Dawley 3. not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

2. TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Age at study initiation: Young adult male and female rats aged between 8 – 12 weeks were used.
- Weight at study initiation: The weight range of approximately 216.9 to 251.9 grams at initiation of dosing.
Body weights at the start : Male Mean: 241.18 g (= 100 %); Minimum : 235.8 g (- 2.23 %); Maximum : 251.9 g (+ 4.44 %)
Female Mean : 221.08 g (= 100 %); Minimum : 216.9 g (- 1.89 %); Maximum : 228.5 g (+ 3.36 %)
- Identification: Each rat was individually identified by the cage number.
- Housing: The rats were individually housed in polycarbonate cages with paddy husk as bedding.
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences, Pune, was provided ad libitum from individual feeders.
- Water (e.g. ad libitum): Water was provided ad libitum from individual bottles attached to the cages. All water was from a local source and passed through the reverse osmosis membrane before use.
- Acclimation period: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.7 to 21.8 degree centigrade.
- Humidity (%): 56.2% to 60.1%
- Air changes (per hr): Ten to fifteen air changes per hour.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

IN-LIFE DATES: 11-07-2017 to 26-07-2017
3. not specified

Type of coverage:

other: 2. semiocclusive 3. Dermal

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

2. TEST SITE
- Area of exposure: Trunk (dorsal surface and sides from scapular to pelvic area)
- % coverage: Approximately 10% of the body surface area.
- Type of wrap if used: Porous gauze dressing and non-irritating tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Distilled water was used to remove residual test item.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
3. not specified

Duration of exposure:

2. 24 hours
3. not specified

Doses:

2. A single dose of 2000 mg of the test item per kilogram of body weight was administered to ten rats (five males and five females).
3. 3000 mg/kg bw

No. of animals per sex per dose:

2. 10 (5/sex).
3. not specified

Control animals:

not specified

Details on study design:

2. - Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Twice daily
- Necropsy of survivors performed: Yes
- Other examinations performed: Clinical Observations and General Appearance: Animals were observed for clinical signs, mortality, until sacrifice.
Onset, duration and severity of any sign were recorded. The clinical signs and mortality observations were conducted at 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily thereafter for 14 day. Daily observation was done as far as possible at the same time.
The observations were included general clinical signs, observations of eyes, mucous membranes, respiratory, circulatory system and behavior pattern.

Evaluation of Dermal Reaction: Dermal reaction was observed daily for study period of 14 days.

Body weights: Individual animal body weights were recorded pre-test (prior to administration of the test item), day 7 and at termination on day 14.

Gross Pathology: Necropsy was performed on animals surviving at the end of the study. Macroscopic examination of all the orifices, cavities and tissues were made and the findings were recorded. All animals surviving the study period were sacrificed by the carbon dioxide asphyxiation technique (day 15).

Histopathology: No gross abnormalities were observed in animals sacrificed terminally hence, no histopathology was performed.
3. not specified

Statistics:

2. not specified
3. not specified

Preliminary study:

2. not specified
3. not specified

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Sex:

not specified

Dose descriptor:

LD50

Effect level:

> 3 000 mg/kg bw

Based on:

test mat.

Mortality:

2. Sex : Male Group I - All animals survived through the study period of 14 days.
Sex : Female Group I - All animals survived through the study period of 14 days.
3. No mortality was observed at 3000 mg/kg bw.

Clinical signs:

other: 2. Sex : Male Group I - Animal treated at the dose level of 2000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days. Sex : Female Group I - Animal treated at the dose level of 2000 mg/kg body weight did not resul

Gross pathology:

2. Gross pathological examination did not reveal any abnormalities in animals from 2000 mg/kg dose group.
3. not specified

Other findings:

2. - Other observations: Evaluation of Dermal Reaction
Sex : Male Group I - Animal treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Sex : Female Group I - Animal treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
3. not specified

Interpretation of results:

other: Not classified

Conclusions:

According to CLP regulation, the given test chemical cannot be classified for acute dermal toxicity, as the LD50 value is >2000 mg/kg bw.

Executive summary:

In different studies, the given test chemical has been investigated for acute dermal toxicity to a greater or lesser extent. Often are the studies based on in-vivo experiments in rodents, i.e. most commonly in rats for test chemical. The studies are summarized as below –

 

The reported study was mentioned in study report and conducted to determine acute dermal toxicity dose by using the given test chemical as per OECD Guideline 402 (Acute Dermal Toxicity) in Sprague Dawley rats.

The test item was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days. Animals exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment.

Hence, the acute dermal LD50 value was considered to be >2000 mg/kg bw, when male and female Sprague Dawley rats were semiocclusively treated with test chemical  by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).

 

The above study is supported with the study mentioned in authoritative database and conducted to determine acute dermal toxicity dose of the given test chemical in rats at the concentration of 3000 mg/kg bw. No mortality was observed in treated rats at 3000 mg/kg bw. Therefore, LD50 was considered to be >3000 mg/kg bw, when rats were treated with test chemical by dermal application.

 

Thus, based on the above summarised studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute dermal toxicity.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Data is from a Klimisch 2 datasource and provides a robust study summary.

Additional information

Acute oral toxicity:

In different studies, the given test chemical has been investigated for acute oral toxicity to a greater or lesser extent. Often are the studies based on in-vivo experiments in rodents, i.e. most commonly in rats for test chemical. The studies are summarized as below –

 

The reported study was mentioned in peer-reviewed journal and conducted to determine acute oral toxicity dose by using the given test chemical in adult Wistar rat at the dose concentration of 2000 mg/kg bw orally. Animals were observed for mortality. No mortality was observed at 2000 mg/kg bw. Hence, the Lethal concentration value (LD50) was considered to be >2000 mg/kg bw, when adult Wistar rat was treated with the given test chemical orally.

 

The above study is supported with another study mentioned in handbook and authoritative database for the given test chemical. The acute oral toxicity study was performed in rat at the dose concentration of 2000 mg/kg bw. Animals were observed for mortality. No mortality was observed at 2000 mg/kg bw. Hence, the Lethal concentration value (LD50) was considered to be >2000 mg/kg bw, when rat was treated with the given test chemical orally.

 

Both the above studies are further supported with the data available in handbook and authoritative database using the given test chemical in rat at the dose concentration of 2000 mg/kg bw. The animals were observed for mortality. No mortality was observed at 2000 mg/kg bw in treated animals. Hence, the Lethal concentration value (LD50) was considered to be >2000 mg/kg bw, when rat was treated with the given test chemical orally.

 

Thus, based on the above summarised studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral toxicity.

 

Acute Inhalation Toxicity:

The acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to very low vapour pressure of the test chemical. Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver. 

 

Acute Dermal Toxicity:

In different studies, the given test chemical has been investigated for acute dermal toxicity to a greater or lesser extent. Often are the studies based on in-vivo experiments in rodents, i.e. most commonly in rats for test chemical. The studies are summarized as below –

 

The reported study was mentioned in study report and conducted to determine acute dermal toxicity dose by using the given test chemical as per OECD Guideline 402 (Acute Dermal Toxicity) in Sprague Dawley rats. The test chemical was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days. Animals exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment. Hence, the acute dermal LD50 value was considered to be >2000 mg/kg bw, when male and female Sprague Dawley rats were semiocclusively treated with test chemical  by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).

 

The above study is supported with the study mentioned in authoritative database and conducted to determine acute dermal toxicity dose of the given test chemical in rats at the concentration of 3000 mg/kg bw. No mortality was observed in treated rats at 3000 mg/kg bw. Therefore, LD50 was considered to be >3000 mg/kg bw, when rats were treated with test chemical by dermal application.

 

Thus, based on the above summarised studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute dermal toxicity.

Justification for classification or non-classification

Based on the above studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw, for acute oral and acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral and acute dermal toxicity. For acute inhalation toxicity wavier was added so, not possible to classify.