Registration Dossier

Administrative data

Description of key information

Acute oral toxicity: 

The acute oral toxicity dose (LD50) for Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate was considered based on data available for the structurally and functionally similar read across chemicals. The LD50 value is between 50 – ≤ 300 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate can be classified as “Category 3” for acute oral toxicity. 

Acute Inhalation toxicity: 

Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate has very low vapour pressure (1.42E-10 Pa), so the potential for the generation of inhalable vapours is very low. Also the normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be highly unlikely and therefore this end point was considered for waiver.

Acute Dermal toxicity:

The acute dermal toxicity dose (LD50) for Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate was considered based on data available for the structurally and functionally similar read across chemicals. The LD50 value is >2000 mg/kg bw.Thus, comparing this value with the criteria of CLP regulation, Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate 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 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:
read-across source
Reason / purpose:
read-across source
Qualifier:
according to
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 the test material (IUPAC name): Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate- Common name: Basic Violet 4- Molecular weight: 515.7375- Molecular formula: C33H45N3O2- Substance type: Organic- Physical Appearance: greenish lustrous liquid- Stability: Stable in recommended storage conditions- Storage Conditions: Ambient Temp (23 to 27 °C)
Species:
rat
Strain:
other: 1. Sprague-Dawley 2. not specified
Sex:
female
Details on test animals 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 197.3 to 204.4 grams.Body weights at the start : Female Mean: 200.28 g (= 100 %); Minimum : 197.3 g (- 1.49 %); Maximum : 204.4 g (+ 2.06 %)- 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 10-07-20172. not specified
Route of administration:
other: 1. oral: gavage 2. oral: unspecified
Vehicle:
other: 1. Polyethylene Glycol - 400 2. not specified
Details on oral exposure:
1. VEHICLE- Concentration in vehicle: 300 mg/kg, 300 mg/kg, 50 mg/kg and 50 mg/kgMAXIMUM DOSE VOLUME APPLIED: 10 ml/kg body weight.2. not specified
Doses:
1. Dose Group I : 300 mg/kg; Dose Group I : 300 mg/kg; Dose Group II : 50 mg/kg; Dose Group II : 50 mg/kg2. 200 mg/kg
No. of animals per sex per dose:
1. Three females were used at each step. 2. not specified
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 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: Gross pathological examination revealed stomach, small and large intestine distended with yellowish liquid ingesta and no gross abnormality observed except colouration hence, no organ collected for histopathology.2. - Other examinations performed: Animals were observed for mortality and clinical signs.
Statistics:
1. No data2. not specified
Preliminary study:
1. No data2. not specified
Sex:
female
Dose descriptor:
LD50
Effect level:
300 mg/kg bw
Based on:
test mat.
Remarks on result:
other: 50% mortality was observed
Sex:
not specified
Dose descriptor:
LD50
Effect level:
> 200 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality was observed
Mortality:
1. Group I Step I : Animals treated at the dose level of 300 mg/kg body weight: One animal died at 4 hours after the dosing. Group I Step II :Animals treated at the dose level of 300 mg/kg body weight: Two animals died at 1 hour after the dosing. Group II Step I :Animals treated at the dose level of 50 mg/kg body weight: All animals survived through the study period of 14 days. Group II Step II :Animals treated at the dose level of 50 mg/kg body weight: All animals survived through the study period of 14 days.2. No mortality was observed at 200 mg/kg bw
Clinical signs:
1. Group I Step I :Animals treated at the dose level of 300 mg/kg body weight resulted in diarrhoea, reduced locomotor activity, ataxic gait and convulsions with onset at 30 minutes to 2 hours after the dosing. All surviving animals were free of signs of toxicity on day 1 after the dosing.Group I Step II :Animals treated at the dose level of 300 mg/kg body weight resulted in diarrhoea, reduced locomotor activity, ataxic gait and convulsions with onset at 30 minutes after the dosing. The only surviving animal was free of signs of toxicity on day 1 after the dosing.Group II Step I :Animals treated at the dose level of 50 mg/kg body weight resulted in diarrhoea, piloerection and ataxic gait with onset at 4 to 6 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 50 mg/kg body weight resulted in diarrhoea and ataxic gait with onset at 4 to 6 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.2. Clinical signs were observed as follows, in gastrointestinal tract - hypermotility, diarrhea; Behavioral changes - convulsions or effect on seizure threshold, and muscle weakness.
Body weight:
1. Group I Step I (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 5.10% and 11.08% respectively. Group I Step II (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 3.99% and 10.21% respectively. Group II Step I (50 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 5.53% and 12.55% respectively. Group II Step II (50 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 6.36% and 12.87% respectively. 2. not specified
Gross pathology:
1. Gross pathological examination did not reveal any abnormalities in animals sacrificed terminally from 50 mg/kg and 300 mg/kg dose groups.Gross pathological examination revealed stomach, small and large intestine distended with yellowish liquid ingesta in found dead animals from 300 mg/kg dose group.2. not specified
Other findings:
1. No data available2. not specified
Interpretation of results:
Category 3 based on GHS criteria
Conclusions:
According to CLP regulation the test chemical Reaction mass of Methylium, tris[4-(diethylamino)phenyl]- & acetate can be classified as "Category 3” for acute oral toxicity, as the LD50 value is in the dose range of 50 – ≤ 300 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 Methylium, tris[4-(diethylamino)phenyl]- & acetate. The studies are as mentioned below:

1. 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 diarrhoea, reduced locomotor activity, ataxic gait and convulsions with onset at 30 minutes to 2 hours after the dosing. One animal died at 4 hours after the dosing. As 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, reduced locomotor activity, ataxic gait and convulsions with onset at 30 minutes after the dosing. Two animals died at 1 hour after the dosing. As mortality was observed at 300 mg/kg dose group, hence 3 female animals were treated with the lower dose of 50 mg/kg of the test item (Step - I). Administration of the test item at 50 mg/kg resulted in diarrhoea, piloerection and ataxic gait with onset at 4 to 6 hours after the dosing and no mortality after the dosing. As no mortality was observed at 24 hours after the dosing, additional 3 female animals were treated with the higher dose of 50 mg/kg of the test item (Step - II). Administration of the test item at 50 mg/kg resulted in diarrhoea and ataxic gait with onset at 4 to 6 hours after the dosing and no mortality after the dosing. All animals from 50 mg/kg dose group survived and 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 50 mg/kg and 300 mg/kg dose groups. Gross pathological examination revealed stomach, small and large intestine distended with yellowish liquid ingesta in found dead animals from 300 mg/kg dose group. The acute oral LD50 of test chemical was 300 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 3 (50 – ≤ 300)” criteria of CLP.

2. The acute oral toxicity of test chemical was tested in rats at the dose concentration of 200 mg/kg bw. Animals were observed for mortality and clinical signs. No mortality was observed at 200 mg/kg bw. Clinical signs were observed as follows, in gastrointestinal tract - hyper motility, diarrhoea; Behavioural changes - convulsions or effect on seizure threshold, and muscle weakness. Hence, LD50 value was considered to be >200 mg/kg bw, when rats were treated with test chemical via oral route.

Thus, based on the above summarised studies, Reaction mass of Methylium, tris[4-(diethylamino)phenyl]- & acetate and it’s structurally and functionally similar read across substances, it can be concluded that LD50 value is between 50 – ≤ 300 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate can be classified as “Category 3” for acute oral toxicity.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
300 mg/kg bw
Quality of whole database:
Data is Klimisch 2 and from study report.

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 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:
read-across source
Reason / purpose:
read-across source
Qualifier:
according to
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
Specific details on test material used for the study:
- Name of the test material (IUPAC name): Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate- Common name: Basic Violet 4- Molecular weight: 515.7375- Molecular formula: C33H45N3O2- Substance type: Organic- Physical Appearance: greenish lustrous liquid- Stability: Stable in recommended storage conditions- Storage Conditions: Ambient Temp (23 to 27 °C)
Species:
rat
Strain:
other: 1. Sprague-Dawley 2. Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
1. 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 216.5 to 244.7 grams at initiation of dosing. Body weights at the start : Male Mean : 239.82 g (= 100 %); Minimum : 231.8 g (- 3.34 %); Maximum : 244.7 g (+ 2.03 %)Female Mean : 220.38 g (= 100 %); Minimum : 216.5 g (- 1.76 %); Maximum : 223.4 g (+ 1.37 %)- 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 26-07-20172. 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 215.0 to 254.3 grams at initiation of dosing. Body weights at the start : Male Mean: 246.28 g (= 100 %); Minimum : 240.9 g (- 2.18 %); Maximum : 254.3 g (+ 3.26 %)Female Mean: 219.02 g (= 100 %); Minimum : 215.0 g (- 1.84 %); Maximum : 223.4 g (+ 2.00 %)- 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): 19.6 to 21.6 degree centigrade.- Humidity (%): 55.0% to 58.4%.- 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: 12-06-2017 to 27-06-2017
Type of coverage:
other: 1. semiocclusive 2. semiocclusive
Vehicle:
other: 1. water 2. water
Details on dermal exposure:
1. 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 - For solids, paste formed: Paste2. 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 - For solids, paste formed: Yes
Duration of exposure:
1. 24 hours2. 24 hours
Doses:
1. 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).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).
No. of animals per sex per dose:
1. 10 (5/sex). 2. 10 (5/sex).
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, 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.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.
Statistics:
1. not specified2. not specified
Preliminary study:
1. not specified2. not specified
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality was observed
Mortality:
1. 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.2. Sex : Male Group I - Animal treated at the dose level of 2000 mg/kg body weight: All animals survived through the study period of 14 days.Sex : Female Group I - Animal treated at the dose level of 2000 mg/kg body weight: All animals survived through the study period of 14 days
Clinical signs:
1. 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 result in any signs of toxicity during the study period of 14 days. 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 result in any signs of toxicity during the study period of 14 days.
Body weight:
1. Sex : Male Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 9.53% and 17.80% respectively. Sex : Female Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 4.91% and 9.62% respectively. 2. Sex : Male Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 9.10% and 17.97% respectively. Sex : Female Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 6.25% and 9.81% respectively.
Gross pathology:
1. Gross pathological examination did not reveal any abnormalities in animals from 2000 mg/kg dose group.2. Gross pathological examination did not reveal any abnormalities in animals from 2000 mg/kg dose group.
Other findings:
1. - Other observations: Evaluation of Dermal ReactionSex : 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.2. - Other observations: Evaluation of Dermal ReactionSex : 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.
Interpretation of results:
other: Not classified
Conclusions:
According to CLP regulation the test chemical Reaction mass of Methylium, tris[4-(diethylamino)phenyl]- & acetate cannot be classified for acute dermal toxicity, as the LD50 value is >2000 mg/kg bw.
Executive summary:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the acute dermal toxicity of the test chemical Reaction mass of Methylium, tris[4-(diethylamino)phenyl]- & acetate. The studies are as mentioned below:

1. The study was designed and conducted to determine the acute dermal toxicity profile of test chemical 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. It was concluded that the acute dermal median lethal dose (LD50) of test chemical, when administered to male and female Sprague Dawley rats was considered to be >2000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that test chemical does not classify as an acute dermal toxicant. CLP Classification: “Not classified”.

2. The study was designed and conducted to determine the acute dermal toxicity profile of test chemical 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. It was concluded that the acute dermal median lethal dose (LD50) of test chemical, when administered to male and female Sprague Dawley rats was considered to be >2000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that test chemical does not classify as an acute dermal toxicant. CLP Classification: “Not classified”.

Thus, based on the above summarised studies, Reaction mass of Methylium, tris[4-(diethylamino)phenyl]- & acetate and it’s structurally similar read across substances, it can be concluded that LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, Reaction mass of Methylium, tris[4-(diethylamino)phenyl]- & acetate 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 study report.

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 Methylium, tris[4-(diethylamino)phenyl]- & acetate. The studies are as mentioned below:

1. 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 diarrhoea, reduced locomotor activity, ataxic gait and convulsions with onset at 30 minutes to 2 hours after the dosing. One animal died at 4 hours after the dosing. As 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, reduced locomotor activity, ataxic gait and convulsions with onset at 30 minutes after the dosing. Two animals died at 1 hour after the dosing. As mortality was observed at 300 mg/kg dose group, hence 3 female animals were treated with the lower dose of 50 mg/kg of the test item (Step - I). Administration of the test item at 50 mg/kg resulted in diarrhoea, piloerection and ataxic gait with onset at 4 to 6 hours after the dosing and no mortality after the dosing. As no mortality was observed at 24 hours after the dosing, additional 3 female animals were treated with the higher dose of 50 mg/kg of the test item (Step - II). Administration of the test item at 50 mg/kg resulted in diarrhoea and ataxic gait with onset at 4 to 6 hours after the dosing and no mortality after the dosing. All animals from 50 mg/kg dose group survived and 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 50 mg/kg and 300 mg/kg dose groups. Gross pathological examination revealed stomach, small and large intestine distended with yellowish liquid ingesta in found dead animals from 300 mg/kg dose group. The acute oral LD50 of test chemical was 300 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 3 (50 – ≤ 300)” criteria of CLP.

2. The acute oral toxicity of test chemical was tested in rats at the dose concentration of 200 mg/kg bw. Animals were observed for mortality and clinical signs. No mortality was observed at 200 mg/kg bw. Clinical signs were observed as follows, in gastrointestinal tract - hyper motility, diarrhoea; Behavioural changes - convulsions or effect on seizure threshold, and muscle weakness. Hence, LD50 value was considered to be >200 mg/kg bw, when rats were treated with test chemical via oral route.

Thus, based on the above summarised studies, Reaction mass of Methylium, tris[4-(diethylamino)phenyl]- & acetate and it’s structurally and functionally similar read across substances, it can be concluded that LD50 value is between 50 – ≤ 300 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate can be classified as “Category 3” for acute oral toxicity.

Acute Inhalation toxicity: 

Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate has very low vapour pressure (1.42E-10 Pa), so the potential for the generation of inhalable vapours is very low. Also the normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be highly unlikely and therefore this end point was considered for waiver.

Acute Dermal Toxicity:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the acute dermal toxicity of the test chemical Reaction mass of Methylium, tris[4-(diethylamino)phenyl]- & acetate. The studies are as mentioned below:

1. The study was designed and conducted to determine the acute dermal toxicity profile of test chemical 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. It was concluded that the acute dermal median lethal dose (LD50) of test chemical, when administered to male and female Sprague Dawley rats was considered to be >2000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that test chemical does not classify as an acute dermal toxicant. CLP Classification: “Not classified”.

2. The study was designed and conducted to determine the acute dermal toxicity profile of test chemical 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. It was concluded that the acute dermal median lethal dose (LD50) of test chemical, when administered to male and female Sprague Dawley rats was considered to be >2000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that test chemical does not classify as an acute dermal toxicant. CLP Classification: “Not classified”.

Thus, based on the above summarised studies, Reaction mass of Methylium, tris[4-(diethylamino)phenyl]- & acetate and it’s structurally similar read across substances, it can be concluded that LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, Reaction mass of Methylium, tris[4-(diethylamino)phenyl]- & acetate cannot be classified for acute dermal toxicity.

Justification for classification or non-classification

Based on the above studies, Reaction mass of Methylium, tris[4-(diethylamino)phenyl]- & acetate and it’s structurally and functionally similar read across substances, it can be concluded that LD50 value is between 50 – ≤ 300 mg/kg bw, for acute oral toxicity; and LD50 value is >2000 mg/kg bw, for acute dermal toxicity. Thus, comparing these values with the criteria of CLP regulation, Reaction mass of Methylium, tris[4-​(diethylamino)​phenyl]​-​ & acetate can be classified as “Category 3” for acute oral toxicity and cannot be classified for acute dermal toxicity. For acute inhalation toxicity wavier were added so, not possible to classify.