4,4'-carbonimidoylbis[N,N-diethylaniline] monohydrochloride

Endpoint summary

Description of key information

Acute oral toxicity:

Under the condition of the study, the acute oral LD50 (Cut-off value) of 4, 4’-carbonimidoylbis [N,N-diethylaniline] monohydrochloride (CAS No. 6358-36-7) was 300 mg/kg body weight. Thus, it was concluded that the acute toxicity study of 4, 4’-carbonimidoylbis [N,N-diethylaniline] monohydrochloride (CAS No. 6358-36-7), when administered via oral route in Sprague Dawley rats falls into the “Category 3 (50 – ≤ 300)” criteria of CLP.

Acute inhalation toxicity:
The study need not be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size.

Acute dermal toxicity:

It was concluded that the acute dermal median lethal dose (LD50) of 4, 4’-carbonimidoylbis [N,N-diethylaniline] monohydrochloride (CAS No. 6358-36-7), when administered to male and female Sprague Dawley rats was found to be greater than 2000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that 4, 4’-carbonimidoylbis [N,N-diethylaniline] monohydrochloride (CAS No. 6358-36-7) does not classify as an acute dermal toxicant.  

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:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Data is from experimental study report.

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:

yes

Test type:

acute toxic class method

Limit test:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Test Item: 4, 4’-carbonimidoylbis [N,N-diethylaniline]monohydrochloride (CAS No. 6358-36-7)
- Source of test material: Sustainability Support Services (Europe) AB, Sweden
- Batch No. of test material: KCP/FS/44/17
- Manufacturing Date: January; 2017
- Expiration date of the lot/batch: December; 2017
- Purity test date: No data available
- Consistency: Solid, powder

RADIOLABELLING INFORMATION (not applicable)
- Radiochemical purity: N/A
- Specific activity: N/A
- Locations of the label: N/A
- Expiration date of radiochemical substance: N/A

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Test Item and prepared formulation(s) were stored at ambient temperature.
- Stability under test conditions: No data available
- Solubility and stability of the test substance in the solvent/vehicle: No data available
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: No data available

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Test item was suspended in Polyethylene Glycol - 400. The formulation was prepared fresh on the day of dosing.
- Preliminary purification step (if any): No data available
- Final dilution of a dissolved solid, stock liquid or gel: No data available
- Final preparation of a solid: No data available

FORM AS APPLIED IN THE TEST: No data available

OTHER SPECIFICS:
Safety Precautions: Safety precautions included use of protective clothing, gloves, masks and eye protection (glasses).

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

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 %)
Total No. of animals : 12
- 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 10-07-2017

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

Polyethylene Glycol - 400

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 300 mg/kg, 300 mg/kg, 50 mg/kg and 50 mg/kg
- Amount of vehicle (if gavage): No data available
- Justification for choice of vehicle: No data available
- Lot/batch no. (if required): No data available
- Purity: No data available

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg body weight.

DOSAGE PREPARATION (if unusual): No data available

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: No data available

Doses:

Dose Group I : 300 mg/kg
Dose Group I : 300 mg/kg
Dose Group II : 50 mg/kg
Dose Group II : 50 mg/kg

No. of animals per sex per dose:

Three females were used at each step.

Control animals:

not specified

Details on study design:

- 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.

Statistics:

No data

Sex:

female

Dose descriptor:

LD50

Effect level:

300 mg/kg bw

Based on:

test mat.

Remarks on result:

other: LD50 cut off value

Mortality:

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.

Clinical signs:

other: 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

Gross pathology:

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.

Other findings:

No data available

Table No. I

Summary of Clinical Signs of Toxicity and Mortality

Test System : Sprague Dawley Rat

Sex : Female

Group I :   

Step No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days From - to	Mortality
I	300	Diarrhoea	2	2 3	1 hr. - 2 hrs. 1 hr. - 6 hrs.	1/3
		Convulsions	1	2	2 hrs.
		Reduced locomotor activity	1	2	30 min. - 2 hrs.
		Ataxic gait	3	1,3 2	30 min. - 6 hrs. 30 min. - 2 hrs.

 

Group I :

Step No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days From - to	Mortality
II	300	Diarrhoea	2	4,6	30 min.	2/3
		Convulsions	1	6	30 min.
		Reduced locomotor activity	3	4,6 5	30 min. 30 min. - 6 hrs.
		Ataxic gait	3	4,6 5	30 min. 30 min. - 6 hrs.

 

Group II :

Step No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days From - to	Mortality
I	50	Diarrhoea	2	7,8	6 hrs.	0/3
		Piloerection	1	7	4 hrs.
		Ataxic gait	2	7,8	4 hrs. - 6 hrs.

 

 

Group II :

Step No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days From - to	Mortality
II	50	Diarrhoea	2	11,12	6 hrs.	0/3
		Ataxic gait	2	11,12	4 hrs. – 6 hrs.

 

 

 

 

Table No. II

Mean Body Weight and Percent Body Weight Gain (g)

Test System : Sprague Dawley Rat

Sex : Female

Group I :

Step No.	Dose (mg/kg body weight)		Before Fasting Body weight	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	300	Mean	199.17	210.15	5.10	222.10	5.69	11.08
		± SD	1.38	0.49	0.06	1.98	0.69	0.79

 

 

Group I :

Step No.	Dose (mg/kg body weight)		Before Fasting Body weight	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
II	300	Mean	201.03	210.90	3.99	223.50	5.97	10.21
		± SD	1.57	-	-	-	-	-

 

 

Group II :

Step No.	Dose (mg/kg body weight)		Before Fasting Body weight	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	50	Mean	201.40	212.53	5.53	226.67	6.66	12.55
		± SD	2.75	3.53	0.73	1.46	1.14	1.13

 

 

Group II :

Step No.	Dose (mg/kg body weight)		Before Fasting Body weight	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
II	50	Mean	199.53	212.20	6.36	225.20	6.12	12.87
		± SD	2.68	1.31	0.84	2.54	0.71	1.23

 

 

 

 

 

Table No.III

Summary of Gross Pathological Findings

Test System : Sprague Dawley Rat

Sex : Female

Group I :  

Step No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	300	1, 3	TS	No abnormality detected
		2	FD	Stomach, small and large intestine distended with yellowish liquid ingesta

 

 Group I :

Step No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
II	300	4, 6	FD	Stomach, small and large intestine distended with yellowish liquid ingesta
		5	TS	No abnormality detected

 

 Group II :

Step No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	50	7 - 9	TS	No abnormality detected

 

 Group II :

Step No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
II	50	10 - 12	TS	No abnormality detected

 

 FD = Found dead

 TS = Terminal Sacrifice

Interpretation of results:

Category 3 based on GHS criteria

Conclusions:

Under the condition of the study, the acute oral LD50 (Cut-off value) of 4, 4’-carbonimidoylbis [N,N-diethylaniline] monohydrochloride (CAS No. 6358-36-7) was 300 mg/kg body weight. Thus, it was concluded that the acute toxicity study of 4, 4’-carbonimidoylbis [N,N-diethylaniline] monohydrochloride (CAS No. 6358-36-7), when administered via oral route in Sprague Dawley rats falls into the “Category 3 (50 – ≤ 300)” criteria of CLP.

Executive summary:

The study was designed and conducted to determine the acute oral toxicity profile of 4, 4’-carbonimidoylbis [N,N-diethylaniline] monohydro chloride (CAS No. 6358-36-7) in 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 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, 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 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 three 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 three 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 (Cut-off value) of 4, 4’-carbonimidoylbis [N,N-diethylaniline] monohydrochloride (CAS No. 6358-36-7) was 300 mg/kg body weight. 

Thus, it was concluded that the acute toxicity study of 4, 4’-carbonimidoylbis [N,N-diethylaniline] monohydrochloride (CAS No. 6358-36-7), when administered via oral route in Sprague Dawley rats falls into the “Category 3 (50 – ≤ 300)” criteria of CLP.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

50 mg/kg bw

Quality of whole database:

Data is from K1 study report

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Data is from experimental study report.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Principles of method if other than guideline:

This study was designed to determine the dermal LD50 of the test item (up to 2000 mg/kg) or to establish a non-lethal dose level of 2000 milligram of test item per kilogram of body weight.

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Test Item: 4,4’-carbonimidoylbis [N,N-diethylaniline] monohydrochloride (CAS No. 6358-36-7)
- Source of test material: Sustainability Support Services (Europe) AB
- Batch No.of test material: KCP/FS/44/17
- Manufacturing Date: January; 2017
- Expiration date of the lot/batch: December; 2017
- Purity test date: No data available
- Consistency: Solid, powder

RADIOLABELLING INFORMATION (Not applicable)
- Radiochemical purity: N/A
- Specific activity: N/A
- Locations of the label: N/A
- Expiration date of radiochemical substance: N/A

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient Temperature
- Stability under test conditions: No data available
- Solubility and stability of the test substance in the solvent/vehicle: No data available
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: No data available

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Test item was grounded to fine powder prior to application. The particulates were moistened
with distilled water before application.
- Preliminary purification step (if any):No data available
- Final dilution of a dissolved solid, stock liquid or gel: No data available
- Final preparation of a solid: No data available

FORM AS APPLIED IN THE TEST: Paste

OTHER SPECIFICS:
Safety Precautions : Safety precautions included use of protective clothing, gloves, masks and eye protection (glasses).

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Females nulliparous and non-pregnant: No data available
- 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 %)
Total No. of animals : 5
Female
Mean : 220.38 g (= 100 %)
Minimum : 216.5 g (- 1.76 %)
Maximum : 223.4 g (+ 1.37 %)
Total No. of animals : 5
- Identification: Each rat was individually identified by the cage number.
- Fasting period before study: No data available
- Housing: The rats were individually housed in polycarbonate cages with paddy husk as bedding.
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences, Pune, was provided ad libitum from individual feeders.
- Water (e.g. ad libitum): Water was provided ad libitum from individual bottles attached to the cages. All water was from a local source and passed through the reverse osmosis membrane before use.
- Acclimation period: 5 days.

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

IN-LIFE DATES: 11-07-2017 to 26-07-2017

Type of coverage:

semiocclusive

Vehicle:

water

Remarks:

(Distilled water)

Details on dermal exposure:

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
- Constant volume or concentration used: No data available
- For solids, paste formed: Paste

VEHICLE
- Amount(s) applied (volume or weight with unit): No data available
- Concentration (if solution): No data available
- Lot/batch no. (if required): No data available
- Purity: No data available

Duration of exposure:

24 hours

Doses:

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:

10 (5/sex).

Control animals:

not specified

Details on study design:

- 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:

No data

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

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.

Clinical signs:

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

Gross pathology:

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

Other findings:

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

Sex : Female
Group I -
Animal treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.

Table No. I

Summary of Clinical Signs of Toxicity and Mortality

Test System : Sprague Dawley Rat

Sex : Male

Group  No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days  From - to	Mortality
I	2000	No clinical signs observed	5	1 - 5	Day 0 - Day 14	0/5

 

 

Sex : Female

Group  No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days  From - to	Mortality
I	2000	No clinical signs observed	5	6 - 10	Day 0 - Day 14	0/5

 

 

Table No. II

Summary of Evaluation of Dermal Reaction

Test System : Sprague Dawley Rat

Sex : Male 

Group  No.	Dose mg/kg	Dermal Reaction	Total Number of Animals	Animal Nos.	Period of signs in days  From - to	Mortality
I	2000	No dermal reaction observed	5	1 - 5	Day 0 - Day 14	0/5

 

 

Sex : Female

Group  No.	Dose mg/kg	Dermal Reaction	Total Number of Animals	Animal Nos.	Period of signs in days  From - to	Mortality
I	2000	No dermal reaction observed	5	6 - 10	Day 0 - Day 14	0/5

 

 

Table No.III

Mean Body Weight and Percent Body Weight Gain (g)

Test System : Sprague Dawley Rat

Sex : Male

Group No.	Dose (mg/kg body weight)		Body weight Day 0	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	2000	Mean	239.82	262.68	9.53	282.52	7.55	17.80
		± SD	5.18	6.37	0.42	7.49	0.75	0.91

 

Sex : Female

Group No.	Dose (mg/kg body weight)		Body weight Day 0	Body weight Day 7	% body weight gain day 0-7	Body weight Day 14	% body weight gain day 7- 14	% body weight gain day 0- 14
I	2000	Mean	220.38	231.20	4.91	241.58	4.49	9.62
		± SD	2.58	2.83	0.71	3.51	0.86	1.29

 

 

Table No.IV

Summary of Gross Pathological Findings

Test System : Sprague Dawley Rat

Sex : Male

Group No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	2000	1 - 5	TS	No abnormality detected

 

Sex : Female

Group No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	2000	6 - 10	TS	No abnormality detected

TS = Terminal Sacrifice

      

 

 

Interpretation of results:

other: Not classified

Conclusions:

It was concluded that the acute dermal median lethal dose (LD50) of 4, 4’-carbonimidoylbis [N,N-diethylaniline] monohydrochloride (CAS No. 6358-36-7), when administered to male and female Sprague Dawley rats was found to be greater than 2000 mg/kg body weight.
Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that 4, 4’-carbonimidoylbis [N,N-diethylaniline] monohydrochloride (CAS No. 6358-36-7) does not classify as an acute dermal toxicant.  
CLP Classification: “Not classified”.

Executive summary:

The study was designed and conducted to determine the acute dermal toxicity profile of 4, 4’-carbonimidoylbis [N,N-diethylaniline]mono hydrochloride (CAS No. 6358-36-7) 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 4, 4’-carbonimidoylbis [N,N-diethylaniline] monohydrochloride (CAS No. 6358-36-7), when administered to male and female Sprague Dawley rats was found to be greater than 2000 mg/kg body weight.

Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that 4, 4’-carbonimidoylbis [N,N-diethylaniline]monohydrochloride (CAS No. 6358-36-7) does not classify as an acute dermal toxicant.  

CLP Classification: “Not classified”.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Data is from k1 study report

Additional information

Acute oral toxicity:

The study was designed and conducted to determine the acute oral toxicity profile of 4, 4’-carbonimidoylbis [N,N-diethylaniline] monohydro chloride (CAS No. 6358-36-7) in 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 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, 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 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 three 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 three 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 (Cut-off value) of 4, 4’-carbonimidoylbis [N,N-diethylaniline] monohydrochloride (CAS No. 6358-36-7) was 300 mg/kg body weight. 

Thus, it was concluded that the acute toxicity study of 4, 4’-carbonimidoylbis [N,N-diethylaniline] monohydrochloride (CAS No. 6358-36-7), when administered via oral route in Sprague Dawley rats falls into the “Category 3 (50 – ≤ 300)” criteria of CLP.

Acute inhalation toxicity:
The study need not be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size.

Acute dermal toxicity:

In different studies, 4,4'-carbonimidoylbis(N,N-diethylaniline) hydrochloride has been investigated for acute dermal toxicity to a greater or lesser extent. Often are the studies based in rodents, i.e. most commonly in rat for 4,4'-carbonimidoylbis(N,N-diethylaniline) hydrochloride along with the study available on structurally similar read across substance Undec-10 -enal [Aldehydes C-11 Undecylenic](CAS no 112-45-8).

The study was designed and conducted to determine the acute dermal toxicity profile of 4, 4’-carbonimidoylbis [N,N-diethylaniline]mono hydrochloride (CAS No. 6358-36-7) 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 4, 4’-carbonimidoylbis [N,N-diethylaniline] monohydrochloride (CAS No. 6358-36-7), when administered to male and female Sprague Dawley rats was found to be greater than 2000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that 4, 4’-carbonimidoylbis [N,N-diethylaniline]monohydrochloride (CAS No. 6358-36-7) does not classify as an acute dermal toxicant.  

It is further supported by experimental study conducted by Sustainability Support Services (2015)on structurally similarread across Undec-10 -enal [Aldehydes C-11 Undecylenic] (CAS no 112-45-8),, acute dermal toxicity was evaluated in five male and five female healthy young adult Wistar rats by using Undec-10 -enal [Aldehydes C-11 Undecylenic] as per OECD No. 403.Twenty four hours prior to dermal application of test item, approximately 10% of body surface area of each rat was clipped. A limit dose of 2000 mg/ kg body weight of test item based on the density (0.8417) and latest body weight was applied by single dermal application and observed for 14 days after treatment. On test day 0, calculated amount of test item was applied directly on the intact skin of clipped area of rats; the porous gauze dressing was put on to the intact skin of clipped area. This porous gauze dressing was covered with a non-irritating tape. After the 24-hour application period, the dressings were removed and the skin was gently wiped with distilled water. The skin reactions were assessed. No mortality was observed in any animal till the end of the experimental period.All the animals were observed with normal clinical signs throughout the experimental period.Mean body weight of both males and females were observed with increase on day 7 and 14, as compared to day 0.In addition,the external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality. Therefore, acute dermal median lethal dose of Undec-10-enal [Aldehydes C-11 Undecylenic] was considered to be > 2000 mg/kg body weight. Undec-10-enal [Aldehydes C-11 Undecylenic] (CAS No. – 112-45-8) is being classified as non toxic by dermal exposure.

This is further supported byexperimentalstudy given by Opdykeet al(Food and Cosmetics Toxicology Volume 11, Issue 3, June 1973, Pages 479)on structurally similarread across Undec-10 -enal [Aldehydes C-11 Undecylenic] (CAS no 112-45-8), acute dermal toxicity was evaluated in rabbits by using Undec-10 -enal [Aldehydes C-11 Undecylenic] in the concentration of 5000 mg/kg bw by dermal application. No effect on survival of treated rabbits were observed at 5000 mg/kg bw. Therefore, LD50 was considered to be > 5000 mg/kg bw when rabbits were treated with Undec-10 -enal [Aldehydes C-11 Undecylenic] by dermal application.

Thus, based on the above studies of 4,4'-carbonimidoylbis(N,N-diethylaniline) hydrochloride and its read across substances , it can be concluded that LD50 value is greater than 2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, 4,4'-carbonimidoylbis[N,N-diethylaniline]monohydrochloride can be “Not classified” for acute dermal toxicity.

Justification for classification or non-classification

Based on k1 data for target 4,4'-carbonimidoylbis(N,N-diethylaniline) hydrochloride (CAS no 6358-36-7) and its Structurally similar read across Undec-10 -enal [Aldehydes C-11 Undecylenic] (CAS no 112-45-8) is likely to be hazardous by acute oral and non-hazardous by dermal route of exposure. Thus, comparing this value with the criteria of CLP regulation, 4-methylphenyl acetate can be “Acute toxicity category 3” for acute oral toxicity and "Not classfied" for Acute dermal toxicity.