Reaction mass of 3H-Indolium, 2-[2-[4-[(2-cyanoethyl)methylamino]phenyl]ethenyl]-1,3,3-trimethyl- & acetate

Endpoint summary

Description of key information

Acute oral toxicity: 

The acute oral toxicity dose (LD50) for Reaction mass of 3H-​Indolium, 2-​[2-​[4-​(dimethylamino)​phenyl]​ethenyl]​-​1,​3,​3-​trimethyl-​ & acetate was considered based on data available for the structurally and functionally similar read across chemicals. The LD50 value is between 300-2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, Reaction mass of 3H-​Indolium, 2-​[2-​[4-​(dimethylamino)​phenyl]​ethenyl]​-​1,​3,​3-​trimethyl-​ & acetate can be classified as “Category 4” for acute oral 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 of read across substances

Justification for type of information:

Data for the target chemical is summarized based on the structurally and functionally similar read across 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

Specific details on test material used for the study:

- Name of test material : Reaction mass of 3H-​Indolium, 2-​[2-​[4-​(dimethylamino)​phenyl]​ethenyl]​-​1,​3,​3-​trimethyl-​ & acetate- Molecular formula : C23H28N2O2- Molecular weight : 364.486 g/mol- Substance type: Organic- Physical state: Liquid

Species:

rat

Strain:

other: 1. Sprague-Dawley 2. Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

1. 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 at the commencement of its dosing.- Weight at study initiation: Body weight range was 188.0 to 208.2 grams.Body weights at the start: Female Mean: 198.51 g (= 100 %) Minimum : 188.0 g (- 5.29 %) Maximum : 208.2 g (+ 4.88 %)- Identification: Each female rat was individually identified by the picric acid marking.- 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): 20.1 to 22.3 degree centigrade.- Humidity (%): 55.1% to 59.3%- 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: 21-06-2017 to 08-07-20172. 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 at the commencement of its dosing.- Weight at study initiation: Body weight range was 196.8 to 204.0 grams.Body weights at the start :Female Mean: 199.92 g (= 100 %) Minimum : 196.8 g (- 1.56 %) Maximum : 204.0 g (+ 2.04 %)- Identification: Each female rat was individually identified by the picric acid marking.- 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): 20.1 to 22.1 degree centigrade.- Humidity (%): 56.1% to 61.1%- 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: 10-07-2017 to 28-07-2017

Route of administration:

other: 1. oral: gavage 2. oral: gavage

Vehicle:

other: 1. water 2. water

Details on oral exposure:

1. VEHICLE- Concentration in vehicle: 300 mg/kg, 300 mg/kg and 2000 mg/kgMAXIMUM DOSE VOLUME APPLIED: 10 ml/kg body weight.2. VEHICLE- Concentration in vehicle: 300 mg/kg, 300 mg/kg, 2000 mg/kg and 2000 mg/kgMAXIMUM DOSE VOLUME APPLIED: 10 ml/kg body weight.

Doses:

1. Dose Group I : 300 mg/kg; Dose Group I : 300 mg/kg; Dose Group II : 2000 mg/kg2. Dose Group I : 300 mg/kg; Dose Group I : 300 mg/kg; Dose Group II : 2000 mg/kg; Dose Group II : 2000 mg/kg

No. of animals per sex per dose:

1. Three females were used at each step. 2. Three females were used at each step.

Control animals:

not specified

Details on study design:

1. - 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 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. 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, found dead and sacrificed 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: Gross pathological examination revealed stomach distended with bluish discoloured mucosa and small and large intestine distended with coloured liquid ingesta and no gross abnormality observed except colouration hence, no organ collected for histopathology.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 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. 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, found dead and sacrificed 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. Histopathology: Gross pathological examination revealed stomach, small and large intestine with test item coloured mucosa in animals from 2000 mg/kg dose group, colouration imparted due to the local irritation of the test item and no other systemic abnormality observed hence, no organ collected for histopathology.

Statistics:

1. No data2. No data

Preliminary study:

1. No data2. No data

Sex:

female

Dose descriptor:

LD50

Effect level:

500 mg/kg bw

Based on:

test mat.

Remarks on result:

other: 50% mortality was observed

Mortality:

1. Group I Step I :Animals treated at the dose level of 300 mg/kg body weight: All animals survived through the study period of 14 days.Group I Step II :Animals treated at the dose level of 300 mg/kg body weight: One animal died on day 1 after the dosing. Group II Step I :Animals treated at the dose level of 2000 mg/kg body weight: Three animals died on day 1 after the dosing. 2. Group I Step I :Animals treated at the dose level of 300 mg/kg body weight: All animals survived through the study period of 14 days.Group I Step II :Animals treated at the dose level of 300 mg/kg body weight: All animals survived through the study period of 14 days.Group II Step I :Animals treated at the dose level of 2000 mg/kg body weight: One animal died at 24 hours after the dosing.Group II Step II :Animals treated at the dose level of 2000 mg/kg body weight: One animal died on day 3, one animal died on day 5 and one animal died on day 10 after the dosing.

Clinical signs:

1. Animals treated at the dose level of 300 mg/kg body weight resulted in diarrhoea and test item coloured faeces with onset at 1 to 4 hours after the dosing. All animals survived through the study period of 14 days and were free of signs of toxicity at on day 2 after the dosing. Animals treated at the dose level of 300 mg/kg body weight resulted in diarrhoea, test item coloured faeces and reduced locomotor activity with onset at 30 minutes to 1 hour after the dosing. All surviving animals were free of signs of toxicity on day 2 after the dosing. Animals treated at the dose level of 2000 mg/kg body weight resulted in diarrhoea, test item coloured faeces, reduced locomotor activity and ataxic gait with onset at 1 to 6 hours after the dosing.2. Animals treated at the dose level of 300 mg/kg body weight resulted in distension, diarrhoea, reduced locomotor activity, ataxic gait and test item coloured feces 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. Animals treated at the dose level of 300 mg/kg body weight resulted in distension, diarrhoea, reduced locomotor activity, ataxic gait and test item coloured feces with onset at 1 hour 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. Animals treated at the dose level of 2000 mg/kg body weight resulted in salivation, distension, diarrhoea, reduced locomotor activity, ataxic gait and test item coloured feces with onset at 30 minutes to 6 hours after the dosing. All surviving animals were free of signs of toxicity on day 7 after the dosing. Animals treated at the dose level of 2000 mg/kg body weight resulted in salivation, distension, diarrhoea, reduced locomotor activity, ataxic gait and test item coloured feces with onset at 30 minutes to 4 hours after the dosing.

Body weight:

1. Group I Step I (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 4.71% and 10.94% respectively. Group I Step II (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 6.10% and 14.58% respectively. Group II Step I (2000 mg/kg) - All animals died on day 1, hence, body weight gain could not be calculated. 2. Group I Step I (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 9.07% and 18.34% respectively. Group I Step II (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 8.68% and 16.75% respectively. Group II Step I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 4.77% and 10.96% respectively. Group II Step II (2000 mg/kg) - All animals died, hence, body weight gain could not be calculated.

Gross pathology:

1. Gross pathological examination did not reveal any abnormalities in animals sacrificed terminally from 300 mg/kg dose group.Gross pathological examination revealed stomach distended with bluish discoloured mucosa and small and large intestine distended with coloured liquid ingesta in found dead animals from 300 mg/kg and 2000 mg/kg dose groups.2. Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg dose group.Gross pathological examination revealed stomach, small and large intestine with test item coloured mucosa in animals sacrificed terminally from 2000 mg/kg dose group and stomach distended with test item coloured mucosa and small and large intestine distended with liquid test item coloured ingesta in found dead animals from 2000 mg/kg dose group.

Other findings:

1. No data available2. No data available

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

According to CLP regulation the test chemical Reaction mass of 3H-​Indolium, 2-​[2-​[4-​(dimethylamino)​phenyl]​ethenyl]​-​1,​3,​3-​trimethyl-​ & acetate (CAS no.: 71598-17-9) can be classified as "Category 4" for acute oral toxicity, as the LD50 value is in the dose range of 300-2000 mg/kg bw.

Executive summary:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the acute oral toxicity of the test chemical Reaction mass of 3H-​Indolium, 2-​[2-​[4-​(dimethylamino)​phenyl]​ethenyl]​-​1,​3,​3-​trimethyl-​ & acetate (CAS no.: 71598-17-9). The studies are as mentioned below:

1. The study was designed and conducted to determine the acute oral toxicity profile of given test chemical in Sprague Dawley rats. Initially, 3 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 resulted in diarrhoea and test item coloured faeces with onset at 1 to 4 hours after the dosing and no mortality after the dosing. As no mortality was observed at 24 hours after the dosing, 3 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 resulted in diarrhoea, test item coloured faeces and reduced locomotor activity with onset at 30 minutes to 1 hour after the dosing. One animal died on day 1 after the dosing. One mortality was observed at 300 mg/kg dose group, hence additional 3 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, test item coloured faeces, reduced locomotor activity and ataxic gait with onset at 1 to 6 hours after the dosing. 3 animals died on day 1 after the dosing. All surviving animals from 300 mg/kg dose group exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities in animals sacrificed terminally from 300 mg/kg dose group. Gross pathological examination revealed stomach distended with bluish discoloured mucosa and small and large intestine distended with coloured liquid ingesta in found dead animals from 300 mg/kg and 2000 mg/kg dose groups. The acute oral LD50 of test chemical was 500 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 4 (300 – ≤ 2000)” criteria of CLP.

2. The study was designed and conducted to determine the acute oral toxicity profile of test chemical in Sprague Dawley rats. Initially, 3 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 resulted in distension, diarrhoea, reduced locomotor activity, ataxic gait and test item coloured faeces with onset at 2 hours after the dosing and no mortality at 24 hours after the dosing. As no mortality was observed at 24 hours after the dosing, 3 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 resulted in distension, diarrhoea, reduced locomotor activity, ataxic gait and test item coloured feces with onset at 1 hour after the dosing and no mortality after the dosing. No mortality was observed at 300 mg/kg dose group, hence additional 3 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 salivation, distension, diarrhoea, reduced locomotor activity, ataxic gait and test item coloured feces with onset at 30 minutes to 6 hours after the dosing. One animal died at 24 hours after the dosing. As one mortality was observed at 24 hours after the dosing, additional 3 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 salivation, distension, diarrhoea, reduced locomotor activity, ataxic gait and test item coloured faeces with onset at 30 minutes to 4 hours after the dosing. One animal died on day 3, one animal died on day 5 and one animal died on day 10 after the dosing. All animals from 300 mg/kg dose group survived through the study period of 14 days and four animals died from 2000 mg/kg dose group after the dosing. Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg dose group. Gross pathological examination revealed stomach, small and large intestine with test item coloured mucosa in animals sacrificed terminally from 2000 mg/kg dose group and stomach distended with test item coloured mucosa and small and large intestine distended with liquid test item coloured ingesta in found dead animals from 2000 mg/kg dose group. The acute oral LD50 of test chemical was 500 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 4 (300 – ≤ 2000)” criteria of CLP.

Thus, based on the above summarised studies, Reaction mass of 3H-​Indolium, 2-​[2-​[4-​(dimethylamino)​phenyl]​ethenyl]​-​1,​3,​3-​trimethyl-​ & acetate (CAS no.: 71598-17-9) and it’s structurally and functionally similar read across substances, it can be concluded that LD50 value is between 300-2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, Reaction mass of 3H-​Indolium, 2-​[2-​[4-​(dimethylamino)​phenyl]​ethenyl]​-​1,​3,​3-​trimethyl-​ & acetate can be classified as “Category 4” for acute oral toxicity.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

500 mg/kg bw

Quality of whole database:

Data is Klimisch 2 and from study report.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Acute oral toxicity:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the acute oral toxicity of the test chemical Reaction mass of 3H-​Indolium, 2-​[2-​[4-​(dimethylamino)​phenyl]​ethenyl]​-​1,​3,​3-​trimethyl-​ & acetate. The studies are as mentioned below:

1. The study was designed and conducted to determine the acute oral toxicity profile of given test chemical in Sprague Dawley rats. Initially, 3 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 resulted in diarrhoea and test item coloured faeces with onset at 1 to 4 hours after the dosing and no mortality after the dosing. As no mortality was observed at 24 hours after the dosing, 3 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 resulted in diarrhoea, test item coloured faeces and reduced locomotor activity with onset at 30 minutes to 1 hour after the dosing. One animal died on day 1 after the dosing. One mortality was observed at 300 mg/kg dose group, hence additional 3 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, test item coloured faeces, reduced locomotor activity and ataxic gait with onset at 1 to 6 hours after the dosing. 3 animals died on day 1 after the dosing. All surviving animals from 300 mg/kg dose group exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities in animals sacrificed terminally from 300 mg/kg dose group. Gross pathological examination revealed stomach distended with bluish discoloured mucosa and small and large intestine distended with coloured liquid ingesta in found dead animals from 300 mg/kg and 2000 mg/kg dose groups. The acute oral LD50 of test chemical was 500 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 4 (300 – ≤ 2000)” criteria of CLP.

2. The study was designed and conducted to determine the acute oral toxicity profile of test chemical in Sprague Dawley rats. Initially, 3 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 resulted in distension, diarrhoea, reduced locomotor activity, ataxic gait and test item coloured faeces with onset at 2 hours after the dosing and no mortality at 24 hours after the dosing. As no mortality was observed at 24 hours after the dosing, 3 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 resulted in distension, diarrhoea, reduced locomotor activity, ataxic gait and test item coloured feces with onset at 1 hour after the dosing and no mortality after the dosing. No mortality was observed at 300 mg/kg dose group, hence additional 3 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 salivation, distension, diarrhoea, reduced locomotor activity, ataxic gait and test item coloured feces with onset at 30 minutes to 6 hours after the dosing. One animal died at 24 hours after the dosing. As one mortality was observed at 24 hours after the dosing, additional 3 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 salivation, distension, diarrhoea, reduced locomotor activity, ataxic gait and test item coloured faeces with onset at 30 minutes to 4 hours after the dosing. One animal died on day 3, one animal died on day 5 and one animal died on day 10 after the dosing. All animals from 300 mg/kg dose group survived through the study period of 14 days and four animals died from 2000 mg/kg dose group after the dosing. Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg dose group. Gross pathological examination revealed stomach, small and large intestine with test item coloured mucosa in animals sacrificed terminally from 2000 mg/kg dose group and stomach distended with test item coloured mucosa and small and large intestine distended with liquid test item coloured ingesta in found dead animals from 2000 mg/kg dose group. The acute oral LD50 of test chemical was 500 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 4 (300 – ≤ 2000)” criteria of CLP.

Thus, based on the above summarised studies, Reaction mass of 3H-​Indolium, 2-​[2-​[4-​(dimethylamino)​phenyl]​ethenyl]​-​1,​3,​3-​trimethyl-​ & acetate and it’s structurally and functionally similar read across substances, it can be concluded that LD50 value is between 300-2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, Reaction mass of 3H-​Indolium, 2-​[2-​[4-​(dimethylamino)​phenyl]​ethenyl]​-​1,​3,​3-​trimethyl-​ & acetate can be classified as “Category 4” for acute oral toxicity.

Justification for classification or non-classification

Based on the above studies, Reaction mass of 3H-​Indolium, 2-​[2-​[4-​(dimethylamino)​phenyl]​ethenyl]​-​1,​3,​3-​trimethyl-​ & acetate and it’s structurally and functionally similar read across substances, it can be concluded that LD50 value is between 300-2000 mg/kg bw, for acute oral toxicity. Thus, comparing this value with the criteria of CLP regulation, Reaction mass of 3H-​Indolium, 2-​[2-​[4-​(dimethylamino)​phenyl]​ethenyl]​-​1,​3,​3-​trimethyl-​ & acetate can be classified as “Category 4” for acute oral toxicity.