Reaction mass of Diethyl[4-[[4-(diethylamino)phenyl]phenylmethylene]-2,5-cyclohexadien-1-ylidene]ammonium and acetate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Data is from experimental study report.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

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

Test material

Test material information

Constituent 1

Reference substance name:

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

EC Number:

228-770-5

EC Name:

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

Cas Number:

6358-36-7

Molecular formula:

C21H29N3.ClH

IUPAC Name:

4,4'-carbonimidoylbis(N,N-diethylaniline) hydrochloride

Test material form:

solid: particulate/powder

Details on test material:

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
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Test Item and prepared formulation(s) were stored at ambient temperature.
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.
OTHER SPECIFICS:
Safety Precautions: Safety precautions included use of protective clothing, gloves, masks and eye protection (glasses).

Test animals

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

Administration / exposure

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

Results and discussion

Preliminary study:

no data

Effect levels

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

300 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no 50% mortality was observed

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:

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.

Body weight:

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.

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

Any other information on results incl. tables

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

Applicant's summary and conclusion

Interpretation of results:

Category 3 based on GHS criteria

Conclusions:

The lethal concentration (LD50-Cut-off value) value for acute oral toxicity test was considered to be 300 mg/kg bw,when female Sprague Dawley rats were treated with test chemical orally via gavage according to OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method).

Executive summary:

The acute oral toxicity profile of test chemical 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.Hence, The lethal concentration (LD50-Cut-off value) value for acute oral toxicity test was considered to be 300 mg/kg bw,when female Sprague Dawley rats were treated with test chemical orally via gavage according to OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method).