Methyl 5-nitrohydrogen.isophthalate

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 the low vapour pressure of the test chemical, which is reported to be 0.000272 Pa. 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 and rabbits 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

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

WoE report is based on 2 acute oral toxicity studies as - WoE 2 and WoE 3.
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. Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

2. TEST ANIMALS
- Source: Vivo Bio Tech Ltd, Telangana
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 9 to 11 Weeks
- Weight at study initiation: 142.08 g to 167.18 g
- Identification:By rat accession number. Identification of individual rats was by cage card and turmeric colour body markings. The rat accession number was allotted during the course of the study. The temporary body marking during acclimatization period was done with crystal violet.
- Fasting period before study: rats were fasted for approximately 16 to 18 hours
- Housing:Rats were housed individually in standard polysulfone cages (Size: L 425 x B 266 x H 185 mm), with stainless steel top grill
- Diet (e.g. ad libitum): Rat & Mice pellet feed, ad libitum
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line water filter-cum-purifier, Mumbai, ad libitum
- Acclimation period: The animals were acclimatized five days for G1-FTS, ten days for G1-STS, thirteen days for G2-FTS and sixteen days for G2-STS. Animals were observed once daily during acclimatization period.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 24°C
- Humidity (%): 59 to 67%
- Air changes (per hr): air conditioned with adequate fresh air supply (12.9 to 14.0 air changes/hour).
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle.

IN-LIFE DATES: From: 30 March 2018 To: 12 July 2018
3. not specified

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

2. VEHICLE
- Concentration in vehicle: 300 & 2000 mg/kg body weight (G1 and G2– First and second treatment steps)
- Amount of vehicle (if gavage):10 mL/kg

DOSAGE PREPARATION (if unusual): A required quantity (g) of test item was weighed in mortar and mixed using pestle by adding small volume of Milli-Q water until a uniform suspension was obtained. The mixture was quantitatively transferred to a measuring
cylinder. Further, a small volume of vehicle was added to the mortar and rinsed with the vehicle, all the rinsing was quantitatively transferred into the measuring cylinder. The final volume was made up to the mark with Milli-Q water to get the desired test item concentration (mg/mL).
3. VEHICLE
- Concentration in vehicle: 33% (W/V)
- Amount of vehicle (if gavage): 15 ml/kg
DOSAGE PREPARATION (if unusual): 33% (W/V) aqueous suspension of the test article was administered by oral gavage at a dosing volume of 15 ml/kg of body weight.

Doses:

2. G1 (FTS) - 300 mg/kg
G1 (STS) - 300 mg/kg
G2 (FTS) - 2000 mg/kg
G2 (STS) - 2000 mg/kg
3. 5000 mg/kg bw

No. of animals per sex per dose:

2. G1 (FTS) - 300 mg/kg - 3
G1 (STS) - 300 mg/kg - 3
G2 (FTS) - 2000 mg/kg - 3
G2 (STS) - 2000 mg/kg - 3
3. 10 rats (5 male and 5 female)

Control animals:

not specified

Details on study design:

2. - Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical signs and pre-terminal deaths - At each step, the animals were observed five times on test day 1 (day of administration) i.e. at 30 minutes and four times at hourly intervals and once daily during days 2 to 15 post administration. Observations included changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern. Attention was directed to the observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma and all observed clinical signs were recorded.

- Body weights - The body weights were recorded on test day 1 (pre-administration), day 8 (7 days post administration) and day 15 (14 days post administration).

- Necropsy of survivors performed: yes, the rats surviving to the end of the observation period were euthanised by using isoflurane anaesthesia and subjected to detailed necropsy. Gross
pathological findings were recorded and reported. Microscopic examination was not carried out as no gross pathological changes were observed.
3. - Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed daily for Mortality and general symptoms.
- Necropsy of survivors performed: yes, limited gross necropsy was performed on all animals.
- Other examinations performed: Animals were observed for clinical signs and body weight change.

Statistics:

2. not specified
3. not specified

Preliminary study:

2. not specified
3. not specified

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

2. G1 - [300 mg/kg body weight - Treatment (FTS and STS)]:There were no pre-terminal deaths.
G2 - [2000 mg/kg body weight - Treatment (FTS and STS)]: There were no pre-terminal deaths.
3. No deaths occurred during the study.

Clinical signs:

other: 2. G1 - [300 mg/kg body weight - Treatment (FTS and STS)]: There were no clinical sings observed in any of the rats. G2 - [2000 mg/kg body weight - Treatment (FTS and STS)]: There were no clinical signs observed in any of the rats. 3. Signs observed withi

Gross pathology:

2. There were no gross pathological changes at necropsy.
3. Gross necropsy findings were within normal limits for all animals.

Other findings:

2. not specified
3. 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 acute oral toxicity study was conducted to assess the toxicological profile of the given test chemical in Wistar rats as per OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method).

The dose formulation was prepared by using Milli-Q water and administered as a single oral gavage to overnight fasted (16 to 18 hours) three female rats (G1-FTS) at the dose of 300 mg/kg body weight. There were no clinical signs of toxicity and pre-terminal deaths observed. As all the rats survived at this step, the test was confirmed with three additional female animals with the same dose of 300 mg/kg body weight (G1-STS). There were no clinical signs of toxicity and pre-terminal deaths observed at this step.

Based on the scheme - Annex 2c of the guideline OECD 423, the test was repeated at the dose of 2000 mg/kg body weight (G2-FTS). There were no clinical signs of toxicity observed. Hence, the test was confirmed with three additional female rats at the same dose of 2000 mg/kg body weight (G2-STS) as per the scheme - Annex 2c of the guideline OECD 423. There were no clinical signs of toxicity and pre-terminal deaths observed at this confirmatory test. Hence, the further dosing was stopped.

The rats were observed for mortality and clinical signs for 14 days post treatment. Body weights were recorded prior to dosing on day 1 and again on days 8 and 15. Necropsy was performed for all the rats at termination. All survived rats gained weight during experimental period. There were no gross pathological changes at necropsy.

Based on the results of the present study, the acute oral LD50 value of the given test chemical is >2000 mg/kg bw. Thus, the test item is not classified for acute oral toxicity. CLP criteria "Not classified".

The above study is supported with another study mentioned in authoritative database and secondary report for the given test chemical. The acute oral toxicity study was conducted in 10 Male and female Sprague-Dawley rats at the concentration of 5000mg/kg bw.

The test material was dissolved as 33% (W/V) aqueous suspension and administered by oral gavage at a dosing volume of 15 ml/kg of body weight. Animals were observed for Mortality and general symptoms for 14 days. Limited gross necropsy was performed on all animals. Animals were observed for clinical signs and body weight change.

No deaths occurred during the study. Signs observed within 24 hours following test article administration included irritability, salivation, redness around the nose, discoloration around the mouth, diarrhoea, wet and/or discoloured inguinal fur and discoloured paws. All clinical signs observed were minor in nature and most of the rats appeared normal 48 hours following test article administration. Mean body weights increased during the study. Gross necropsy findings were within normal limits for all animals. Hence, the LD50 value was considered to be >5000 mg/kg bw, when 10 Male and female Sprague-Dawley rats were treated with the given test chemical via oral gavage route.

 

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 report.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

study scientifically not necessary / other information available

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

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 dermal toxicity studies as- WoE 2, WoE 3 and WoE 4.
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:

other: 2. rat 3. rabbit 4. rabbit

Strain:

other: 2. Wistar 3. New Zealand White 4. New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

2. TEST ANIMALS
- Source: Vivo Bio Tech Ltd., Telangana
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8 to 10 weeks
- Weight at study initiation: Females: 213.42 to 248.13 g
- Identification:By rat accession number. Identification of individual rats is by cage card and crystal violet colour body markings. The temporary body marking during acclimatization period was done with crystal violet. The rat accession numbers were allotted during the course of the study and was included in raw data and reported.
- Housing: Animals were housed individually in standard polysulfone cages (Size: L 425 x B 266 x H 185 mm), with stainless steel top grill. Additionally, polycarbonate rat huts were placed inside the cage as enrichment objects and were changed along with the cage once a week. Bedding: Steam sterilized corn cob was used and changed once a week along with the cage.
- Diet (e.g. ad libitum): Rat & Mice pellet feed, ad libitum
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line water filter-cum-purifier, ad libitum
- Acclimation period: The rats were acclimatized for six, eight, twelve and fourteen days before treatment for dose range finding and main study respectively under standard laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 24°C
- Humidity (%): 57 to 67 %
- Air changes (per hr): air conditioned with adequate fresh air supply (12.9 to 14.0 air changes/hour).
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle

IN-LIFE DATES: From: 30 March 2018 To: 29 June 2018
3. not specified
4. TEST ANIMALS
- Age at study initiation: about 3 months
- Weight at study initiation: Animals were weighed an average of 2.59 kg (Male) and 2.45 kg (Female).

Type of coverage:

other: 2. semiocclusive 3. occlusive 4. occlusive

Vehicle:

other: 2. water 3. unchanged (no vehicle) 4. unchanged (no vehicle)

Details on dermal exposure:

2. TEST SITE
- Area of exposure: clipped skin of dorsolateral thoracic surface
- % coverage: 10% of the body surface
- Type of wrap if used: The applied area was covered with cotton gauze (size: Females: 8 x 5 cm of 6 ply) and it was secured in position by adhesive tape wound around the torso.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The dressing was removed and the applied area was washed with deionized water and wiped dry using clean towel.
- Time after start of exposure:24 hours

TEST MATERIAL
- For solids, paste formed: yes, the test item formed a paste in Milli-Q water
3. TEST SITE
- Area of exposure: The test material was applied to the clipped back of each animal.
4. TEST SITE
- Area of exposure: The test material was applied on the back
- Type of wrap if used: Covered with an occlusive wrap.

Duration of exposure:

2. 24 hours
3. 24 hours
4. not specified

Doses:

2. Two treatment group
G1 – DRF and Main
3. 2000 mg/kg bw
4. 2000 mg/kg bw

No. of animals per sex per dose:

2. DRF - 2000 mg/kg - 1
Main - 2000 mg/kg - 2
3. Total = 10 (5/sex)
4. Total = 10 (5/sex)

Control animals:

not specified

Details on study design:

2. - Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical examination and pre-terminal deaths - The animals were observed for clinical signs and pre-terminal deaths (mortality) once during first 30 minutes after application, and at hourly intervals for 6 hours after application on the day of treatment (day 1) and once daily during Days 2 to 15. In addition, treatment site at was observed 24, 48 and 72 hours after removal of test item using the Draize criteria (Refer Annexure 4 of this report). All rats were observed for changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

- Body weights - Individual body weights of animals were recorded on test days 1 (Pre-application), 8 (7 days post application), and 15 (14 days post application).

- Necropsy of survivors performed: yes, at the end of the observation period, all rats were euthanised and exsanguinated under isoflurane anesthesia and subjected to detailed necropsy by an experienced prosector and the findings were recorded. Microscopic examination was not carried out as no gross pathological changes were observed.
3. - Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed daily for Mortality and general symptoms.
- Necropsy of survivors performed: yes
- Other examinations performed: Animals were observed for clinical signs and body weight change.
4. - Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed daily for Mortality and general symptoms; Body weights were assessed at dosing, and on Days 7 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: Animals were observed for clinical signs.

Statistics:

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

Preliminary study:

2. Dose range finding study - As per the available literature, the LD50 of acute dermal median lethal dose of Rabbit is >5000 mg/kg body weight. Hence an intial dose of 2000 mg/kg body weight was tested with 1 female rat (dose range finding study). As there was no mortality at this dose the main study was conducted further with 2 animals to confirm the classification. There was no test item-related mortality. The subsequent dosing was done approximately 48 hours after the previous dosing.
3. not specified
4. not specified

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

2. There were no pre-terminal deaths (mortality) observed during the study.
3. No mortality observed at 2000 mg/kg bw.
4. No mortality observed at 2000 mg/kg bw.

Clinical signs:

other: 2. There were no clinical signs observed during the study. 3. Mild dermal irritation was observed in 4 animals immediately following unwrapping. No adverse treatment-related clinical signs were observed. 4. Clinical signs noted consisted of an erythema at

Gross pathology:

2. No abnormality was detected at necropsy.
3. No gross pathological lesions due to treatment were evident in any of the animals at necropsy.
4. No alterations were noted during gross necropsy.

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 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 and rabbits for test chemical. The studies are summarized as below –

 

The acute dermal toxicity study was conducted as per OECD Guideline 402 (Acute Dermal Toxicity) with the dose concentration of 2000 mg/kg in 1 female for the dose range finding study, followed by additional 2 females for main study at the dose of 2000 mg/kg body weight in Wistar rats.

Based on the individual body weight, the test item at the dose of 2000 mg/kg body weight was weighed on an aluminium foil and made as a paste in Milli-Q water and applied directly to the clipped skin (clipping was done approximately 24 hour prior to application) of the animal to cover about 10% of the body surface of the animal (semi-occlusive). The area of application was covered with cotton gauze (size: Females: 8 x 5 cm of 6 ply) and it was secured in position by adhesive tape wrapped around the torso. The test item contact period with the skin was for 24 hours. After the 24 hours contact period, the adhesive tape and cotton gauge was removed and the applied area was washed with deionized water and wiped dry using clean towels.

All the rats were observed for clinical signs of toxicity and mortality for 14 days post application. There were no clinical signs of toxicity and mortality. There was no skin reaction observed at test item applied area. Body weight was measured on days 1, 8 and 15 and all rats gained weight during experimental period. At the end of observation period, all surviving animals were euthanized and subjected to necropsy. There were no abnormalities detected at the necropsy.

Based on the present study results, the acute dermal LD50 of the given test chemical is >2000 mg/kg body weight in female Wistar rats. The test item does not classify for acute dermal toxicity. CLP classification "Not classified”.

 

The above study is supported with another study available in authoritative database and secondary report and conducted on 10 Male and female New Zealand White rabbits by using the given test chemical at the concentration of 2000 mg/kg bw.

The test material was applied to the clipped back of each animal for 24 hours. Animals were observed daily for Mortality, clinical signs and body weight change.

No mortality observed at 2000 mg/kg bw. Mild dermal irritation was observed in 4 animals immediately following unwrapping. No adverse treatment-related clinical signs were observed. Mean body weights increased during the study. No gross pathological lesions due to treatment were evident in any of the animals at necropsy.

Therefore, the acute dermal LD50 value was considered to be >2000 mg/kg bw, when Male and female New Zealand White rabbits were treated with the given test chemical occlusively to the clipped back skin.

 

Both the above studies are further supported with the study mentioned in authoritative database and secondary reports. The reported study was designed and conducted to determine the acute dermal toxicity profile of the given test chemical. In this study, 10 Male and female New Zealand rabbitswere treated by dermal application at the concentration of 2000 mg/kg bw.

The test material was applied on the back and covered with an occlusive wrap. Prior to application, the backs were shaved and moistened with water. Animals were observed daily for Mortality and general symptoms; Body weights were assessed at dosing, and on Days 7 and 14.

No mortality observed at 2000 mg/kg bw. Clinical signs noted consisted of an erythema at the application site immediately after unwrapping in 2/5 males and 4/5 females. All animals appeared normal by Day 4. Mean body weights increased during the study. No alterations were noted during gross necropsy.

Therefore, the acute dermal LD50 value was considered to be >2000 mg/kg bw, when Male and female New Zealand rabbits were treated with the given test chemical occlusively to the skin.

 

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 Klimisch 2 and from report.

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 acute oral toxicity study was conducted to assess the toxicological profile of the given test chemical in Wistar rats as per OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method).

The dose formulation was prepared by using Milli-Q water and administered as a single oral gavage to overnight fasted (16 to 18 hours) three female rats (G1-FTS) at the dose of 300 mg/kg body weight. There were no clinical signs of toxicity and pre-terminal deaths observed. As all the rats survived at this step, the test was confirmed with three additional female animals with the same dose of 300 mg/kg body weight (G1-STS). There were no clinical signs of toxicity and pre-terminal deaths observed at this step.

Based on the scheme - Annex 2c of the guideline OECD 423, the test was repeated at the dose of 2000 mg/kg body weight (G2-FTS). There were no clinical signs of toxicity observed. Hence, the test was confirmed with three additional female rats at the same dose of 2000 mg/kg body weight (G2-STS) as per the scheme - Annex 2c of the guideline OECD 423. There were no clinical signs of toxicity and pre-terminal deaths observed at this confirmatory test. Hence, the further dosing was stopped.

The rats were observed for mortality and clinical signs for 14 days post treatment. Body weights were recorded prior to dosing on day 1 and again on days 8 and 15. Necropsy was performed for all the rats at termination. All survived rats gained weight during experimental period. There were no gross pathological changes at necropsy.

Based on the results of the present study, the acute oral LD50 value of the given test chemical is >2000 mg/kg bw. Thus, the test item is not classified for acute oral toxicity. CLP criteria "Not classified".

The above study is supported with another study mentioned in authoritative database and secondary report for the given test chemical. The acute oral toxicity study was conducted in 10 Male and female Sprague-Dawley rats at the concentration of 5000mg/kg bw.

The test material was dissolved as 33% (W/V) aqueous suspension and administered by oral gavage at a dosing volume of 15 ml/kg of body weight. Animals were observed for Mortality and general symptoms for 14 days. Limited gross necropsy was performed on all animals. Animals were observed for clinical signs and body weight change.

No deaths occurred during the study. Signs observed within 24 hours following test article administration included irritability, salivation, redness around the nose, discoloration around the mouth, diarrhoea, wet and/or discoloured inguinal fur and discoloured paws. All clinical signs observed were minor in nature and most of the rats appeared normal 48 hours following test article administration. Mean body weights increased during the study. Gross necropsy findings were within normal limits for all animals. Hence, the LD50 value was considered to be >5000 mg/kg bw, when 10 Male and female Sprague-Dawley rats were treated with the given test chemical via oral gavage route.

 

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 the low vapour pressure of the test chemical, which is reported to be 0.000272 Pa. 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 and rabbits for test chemical. The studies are summarized as below –

 

The acute dermal toxicity study was conducted as per OECD Guideline 402 (Acute Dermal Toxicity) with the dose concentration of 2000 mg/kg in 1 female for the dose range finding study, followed by additional 2 females for main study at the dose of 2000 mg/kg body weight in Wistar rats.

Based on the individual body weight, the test item at the dose of 2000 mg/kg body weight was weighed on an aluminium foil and made as a paste in Milli-Q water and applied directly to the clipped skin (clipping was done approximately 24 hour prior to application) of the animal to cover about 10% of the body surface of the animal (semi-occlusive). The area of application was covered with cotton gauze (size: Females: 8 x 5 cm of 6 ply) and it was secured in position by adhesive tape wrapped around the torso. The test item contact period with the skin was for 24 hours. After the 24 hours contact period, the adhesive tape and cotton gauge was removed and the applied area was washed with deionized water and wiped dry using clean towels.

All the rats were observed for clinical signs of toxicity and mortality for 14 days post application. There were no clinical signs of toxicity and mortality. There was no skin reaction observed at test item applied area. Body weight was measured on days 1, 8 and 15 and all rats gained weight during experimental period. At the end of observation period, all surviving animals were euthanized and subjected to necropsy. There were no abnormalities detected at the necropsy.

Based on the present study results, the acute dermal LD50 of the given test chemical is >2000 mg/kg body weight in female Wistar rats. The test item does not classify for acute dermal toxicity. CLP classification "Not classified”.

 

The above study is supported with another study available in authoritative database and secondary report and conducted on 10 Male and female New Zealand White rabbits by using the given test chemical at the concentration of 2000 mg/kg bw.

The test material was applied to the clipped back of each animal for 24 hours. Animals were observed daily for Mortality, clinical signs and body weight change.

No mortality observed at 2000 mg/kg bw. Mild dermal irritation was observed in 4 animals immediately following unwrapping. No adverse treatment-related clinical signs were observed. Mean body weights increased during the study. No gross pathological lesions due to treatment were evident in any of the animals at necropsy.

Therefore, the acute dermal LD50 value was considered to be >2000 mg/kg bw, when Male and female New Zealand White rabbits were treated with the given test chemical occlusively to the clipped back skin.

 

Both the above studies are further supported with the study mentioned in authoritative database and secondary reports. The reported study was designed and conducted to determine the acute dermal toxicity profile of the given test chemical. In this study, 10 Male and female New Zealand rabbitswere treated by dermal application at the concentration of 2000 mg/kg bw.

The test material was applied on the back and covered with an occlusive wrap. Prior to application, the backs were shaved and moistened with water. Animals were observed daily for Mortality and general symptoms; Body weights were assessed at dosing, and on Days 7 and 14.

No mortality observed at 2000 mg/kg bw. Clinical signs noted consisted of an erythema at the application site immediately after unwrapping in 2/5 males and 4/5 females. All animals appeared normal by Day 4. Mean body weights increased during the study. No alterations were noted during gross necropsy.

Therefore, the acute dermal LD50 value was considered to be >2000 mg/kg bw, when Male and female New Zealand rabbits were treated with the given test chemical occlusively to the skin.

 

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.