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Description of key information

Acute oral toxicity:

The acute oral median lethal dose (LD50) of the test chemical was >2000 mg/kg body weight. Thus, it can be concluded that the test chemical is likely to fall in category "Not classified" when administered by oral route of exposure, as per the criteria mentioned in CLP regulation.

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:
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 for the target chemical is summarized based on data from various test chemicals
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Principles of method if other than guideline:
The purpose of this study was to assess the Toxicological profile of test item to a single administration via oral route to Sprague Dawley rats. This study was designed to determine the acute toxicity at fixed dose levels by oral route of the test item.
GLP compliance:
not specified
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
other: 2. Sprague-Dawley ; 3. Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
2. TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Females nulliparous and non-pregnant: yes
- Age at study initiation: Female rats of the age of approximately 8 to 12 weeks old were used.
- Weight at study initiation: Body weight range was 199.1 to 219.9 grams.
Body weights at the start : Female Mean: 206.81 g (= 100 %); Minimum : 199.1 g (- 3.73 %); Maximum : 219.9 g (+ 6.33 %)
- Fasting period before study: Approximately 16 hours or more.
- Housing: The rats were housed in polycarbonate cages.
- 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): 20.6 to 23.2 degree centigrade.
- Humidity (%): 55.1% to 58.6%.
- 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: 26-09-2016 to 15-10-2016

3. Species:Rattus norvegicus (Rat)
Strain:Wistar
Number and Sex: Six Females
Supplier / Source: In-house animals, bred.
Health Status :Healthy young adult animals were used for the study. Females were nulliparous and non pregnant.
Body weight of animals:Minimum: 141 g Maximum: 171 g (Individual body weights were within ± 20 % prior to treatment after overnight fasting)
Age: 08 - 11 weeks at the time of dosing.
Acclimatisation:Animal nos. 1-3 were acclimatized for 6 days and 4-6 for 10 days, prior to administr ation of the test item.
Identification :The animals were marked temporarily on tail, permanently on toe pad micro tattooing and cage cards. Individual cage cards were labelled with study no., study type, test system, group, dose, sex, animal number experimental start and completion date. Husbandry Conditions
Diet:All animals were provided conventional laboratory rodent diet.
Bedding:All cages were provided with corn cobs (Sparconn Life Sciences Bangalore) SPAR – 27 /2014.
Water: Aqua guard filtered tap water was provided ad libitum via drinking bottles.
Husbandry:The animals were housed individually in polycarbonate cages.
Room Sanitation:The experimental room floor and work tops were swept and mopped with dis infectant solution every day.
Cages and water bottle:All the cages and water bottles were changed at least twice every week.
Experimental Room Condition
Temperature:Minimum: 19.40 °C Maximum: 22.10 °C
Relative humidity:Minimum: 46.60% Maximum: 65.40%
Light-dark-rhythm: 12 hour light and 12 hour dark
Air Changes: More than 12 changes per hour

Route of administration:
other: oral: 1. Gavage; 2. Unspecified
Vehicle:
water
Remarks:
(Distilled water)
Details on oral exposure:
2. VEHICLE
- Concentration in vehicle: 300 mg/kg, 300 mg/kg, 2000 mg/kg and 2000 mg/kg
MAXIMUM DOSE VOLUME APPLIED: 10 ml per kg of body weight.

3. Distilled water was selected as a vehicle based on solubility testing.
Doses:
2. Dose Group I : 300 mg/kg
Dose Group I : 300 mg/kg
Dose Group II : 2000 mg/kg
Dose Group II : 2000 mg/kg

3. 2000 mg/kg bw
No. of animals per sex per dose:
2. Three females were used at each step.
3. 2000 mg/kg bw- 6 female rats
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 and morbidity, until sacrifice.
Onset, duration and severity of any sign were recorded. The clinical signs and mortality observations were conducted at immediately (0 to 5 minutes), 5, 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily therea fter for 14 day. Daily observation was done as far as possible at the same time.
Body weights: Individual animal body weights were recorded, before fasting, prior to administration of the test item (fasting body weights), weekly thereafter and at termination on day 14. Weight changes were calculated and recorded.
Gross Pathology: Necropsy was performed on all animals at the end of the study period on day 15. 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.
Histopathology: No gross abnormalities were observed in animals sacrificed terminally hence, no histopathology was performed.

3. - Duration of observation period following administration:
14 days - Frequency of observations and weighing: daily
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, mortality, body weight, pathology
Mortality: All surviving animals were observed twice daily (morning and evening) for morbidity and mortality, throughout the acclimatization and study period
Clinical signs: After test item administration, individual animals were frequently observed at 30 m inutes, 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing). Subsequently, all surviving animals were observed once a day during the 14 day observation period.
Body weight: All surviving rats were weighed on days 0 (prior to dosing), 7 and 14.
Gross pathology: At the end of 14 day observation period, all the survived rats were euthanised by overdose of CO2. All the animals were observed for external and internal gross pathology.
Statistics:
No data
Preliminary study:
No data
Sex:
female
Dose descriptor:
LD50
Remarks:
2
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality was obseved
Sex:
female
Dose descriptor:
LD50
Remarks:
3
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality was observed
Mortality:
2. All animals treated at the dose level of 300 mg/kg body weight and 2000mg/kg body weight survived through the study period of 14 days.
3. No mortality was observed in the animals treated with 2000 mg/Kg dose throughout the 14 days observation period
Clinical signs:
2. Group I Step I : Animals treated at the dose level of 300 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days.
Group I Step II : Animals treated at the dose level of 300 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days.
Group II Step I : Animals treated at the dose level of 2000 mg/kg body weight resulted in diarrhoea (black colour stools) in all animals with onset at 2 hours after the dosing. All animals survived through the study period of 14 days and were free of signs of toxicity on day 1 after the dosing.
Group II Step II : Animals treated at the dose level of 2000 mg/kg body weight resulted in diarrhoea (black colour stools) in all animals with onset at 4 hours after the dosing. All animals survived through the study period of 14 days and were free of signs of toxicity on day 1 after the dosing.
Staining of the stool is attributed to the black colour of the test item.

3. At 2000 mg/Kg bw, all the animals were normal thorughout the experimenatl period
Body weight:
2. Group I Step I (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 4.45% and 12.76% respectively.
Group I Step II (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 3.56% and 12.49% respectively.
Group II Step I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 3.06% and 10.98% respectively.
Group II Step II (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 2.17% and 9.86% respectively.

3. Mean body weight of all surviving animals treated with 2000 mg/kg body weight was observed with gain on day 7 and 14, as compared to day 0
Gross pathology:
2. Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups.
3. No external and internal gross pathology changes were seen in all the six animals treated with 2000 mg/Kg bw during terminal sacrifice
Other findings:
No data
Interpretation of results:
other: Not classified
Conclusions:
Under the condition of the study, the acute oral LD50 of the test chemical was >2000 mg/kg body weight. Thus, it was concluded that the acute toxicity study of test chemical, when administered via oral route in rats falls into the “Category Not classified” criteria of CLP.
Executive summary:

Data available for the various test chemicals was reviewed to determine the toxic nature of the test chemical. The studies are as mentioned below:

The acute oral toxicity study was designed and conducted as per OECD 423 for the test chemical using Sprague Dawley rats. Initially, three female animals were treated at the dose level of 300 mg/kg body weight of the test item (Step - I). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality at 24 hours after the dosing. As no mortality was observed at 24 hours after the dosing, three female animals were added to the study and treated with the same dose of 300 mg/kg of the test item (Step - II). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality after the dosing. No mortality was observed at 300 mg/kg dose group, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - I). Administration of the test item at 2000 mg/kg resulted in diarrhoea (black colour stools) in all animals with onset at 2 hours and no mortality after the dosing. As no mortality were observed at 24 hours after the dosing, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - II). Administration of the test item at 2000 mg/kg resulted in diarrhoea (black colour stools) in all animals with onset at 4 hours and no mortality after the dosing. All animals from 300 mg/kg and 2000 mg/kg dose groups survived through the study period of 14 days. Staining of the stool is attributed to the black colour of the test item. Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups. The acute oral LD50 of test chemical was >2000 mg/kg body weight. Thus, it was concluded that the acute toxicity study of test chemical, when administered via oral route in Sprague Dawley rats falls into the “Category Not classified” criteria of CLP.

An acute toxicity study as per OECD 423 guideline was conducted to examine the effects of the test chemical through a single exposure. Female wistar rats were used for this experiment. The starting dose was selected as 2000 mg/kg bw. All surviving animals were observed twice daily (morning and evening) for morbidity and mortality, throughout the acclimatization and study period. After test item administration, individual animals were frequently observed at 30 minutes, 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing). Subsequently, all surviving animals were observed once a day during the 14 day observation period. All surviving rats were weighed on days 0 (prior to dosing), 7 and 14. At the end of 14 day observation period, all the survived rats were euthanized by overdose of CO2. All the animals were observed for external and internal gross pathology. However, no mortality or any other changes were observed at any dose group. Based on all the observations and results, it was concluded that the acute oral medial lethal dose (LD50) of the test chemical was >2000 mg/kg bw and thus was not acutly toxic to the female Wistar rats.

Based on the data available and applying the weight of evidence approach, the acute oral median lethal dose (LD50) of the test chemical was >2000 mg/kg body weight. Thus, it was concluded that the acute toxicity study of test chemical, when administered via oral route in rats falls into the “Category -Not classified” criteria of CLP.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Data is from K2 source

Additional information

Acute oral toxicity:

Data available for the various test chemicals was reviewed to determine the toxic nature of the test chemical. The studies are as mentioned below:

The acute oral toxicity study was designed and conducted as per OECD 423 for the test chemical using Sprague Dawley rats. Initially, three female animals were treated at the dose level of 300 mg/kg body weight of the test item (Step - I). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality at 24 hours after the dosing. As no mortality was observed at 24 hours after the dosing, three female animals were added to the study and treated with the same dose of 300 mg/kg of the test item (Step - II). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality after the dosing. No mortality was observed at 300 mg/kg dose group, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - I). Administration of the test item at 2000 mg/kg resulted in diarrhoea (black colour stools) in all animals with onset at 2 hours and no mortality after the dosing. As no mortality were observed at 24 hours after the dosing, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - II). Administration of the test item at 2000 mg/kg resulted in diarrhoea (black colour stools) in all animals with onset at 4 hours and no mortality after the dosing. All animals from 300 mg/kg and 2000 mg/kg dose groups survived through the study period of 14 days. Staining of the stool is attributed to the black colour of the test item. Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups. The acute oral LD50 of test chemical was >2000 mg/kg body weight. Thus, it was concluded that the acute toxicity study of test chemical, when administered via oral route in Sprague Dawley rats falls into the “Category Not classified” criteria of CLP.

An acute toxicity study as per OECD 423 guideline was conducted to examine the effects of the test chemical through a single exposure. Female wistar rats were used for this experiment. The starting dose was selected as 2000 mg/kg bw. All surviving animals were observed twice daily (morning and evening) for morbidity and mortality, throughout the acclimatization and study period. After test item administration, individual animals were frequently observed at 30 minutes, 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing). Subsequently, all surviving animals were observed once a day during the 14 day observation period. All surviving rats were weighed on days 0 (prior to dosing), 7 and 14. At the end of 14 day observation period, all the survived rats were euthanized by overdose of CO2. All the animals were observed for external and internal gross pathology. However, no mortality or any other changes were observed at any dose group. Based on all the observations and results, it was concluded that the acute oral medial lethal dose (LD50) of the test chemical was >2000 mg/kg bw and thus was not acutly toxic to the female Wistar rats.

Based on the data available and applying the weight of evidence approach, the acute oral median lethal dose (LD50) of the test chemical was >2000 mg/kg body weight. Thus, it can be concluded that the test chemical is likely to fall in category "Not classified" when administered by oral route of exposure, as per the criteria mentioned in CLP regulation.

Justification for classification or non-classification

Based on the data available and applying the weight of evidence approach, it can be concluded that the test chemical is likely to fall in category "Not classified" when administered by oral route of exposure, as per the criteria mentioned in CLP regulation.