N,N'-[1,4-phenylenebis(methylene)]bis{N,N-dimethyl-3-[(prop-2-enoyl)amino]propan-1-aminium} dichloride

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

04 - 20 Feb 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

Adopted in 2001

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Slovak National Accreditation Service, Bratislava, Slovak Republic

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

{[4-({dimethyl[3-(prop-2-enamido)propyl]azaniumyl}methyl)phenyl]methyl}dimethyl[3-(prop-2-enamido)propyl]azanium dichloride

EC Number:

812-548-5

Cas Number:

1621424-03-0

Molecular formula:

C24H40Cl2N4O2

IUPAC Name:

{[4-({dimethyl[3-(prop-2-enamido)propyl]azaniumyl}methyl)phenyl]methyl}dimethyl[3-(prop-2-enamido)propyl]azanium dichloride

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: VELAZ PRAHA, Czech Republic
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: 8 - 12 weeks of age at first dose
- Weight at study initiation: 185 - 211 g (range)
- Fasting period before study: Animals were fasted 10 - 12 h prior to dosing (food but not water was withheld over night). After test substance administration, food was withheld for a further 3 - 4 h.
- Housing: Animals were housed in plastic cages suspended on stainless steel racks. 3 animals per cage. Bedding used was SAFE 3/4 S, JRS J. Rettenmaier & Söhne GmbH + Co KG.
- Diet: laboratory food ssniff (ssniff Spezialdiäten GmbH, Germany), ad libitum
- Water: tap water for human consumption, ad libitum
- Acclimation period: 5 days prior to start of treatment

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 50 - 60
- Photoperiod (hrs dark / hrs light): 12 /12

IN-LIFE DATES: From: 04 Feb 2020 To: 20 Feb 2020

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Water for injection

Details on oral exposure:

VEHICLE
- Amount of vehicle: 5 mL/kg bw
- Justification for choice of vehicle: An aqueous solution/suspension/emulsion is a common vehicle in toxicity studies like OECD TG 423.
- Lot/batch no.: 18H0902 (Bieffe Medital S. p. A, Italy)
- Purity: water for injection

MAXIMUM DOSE VOLUME APPLIED: 5 mL/kg bw

DOSAGE PREPARATION: The required amount of the test substance was mixed with the vehicle shortly before administration.

CLASS METHOD
- Rationale for the selection of the starting dose: The limit dose of 2000 mg/kg bw was used as a starting dose because available information indicated that the test substance was likely to be non-toxic regarding acute toxicity.

Doses:

2000 mg/ kg bw

No. of animals per sex per dose:

6 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed for clinical signs individually immediatelly, 0.5, 1, 2, and 4 h post administration. Each animal was observed daily for the next 14 days for clinical signs of toxicity. Individual body weights of animals were measured immediately before test substance administration and weekly thereafter.
- Necropsy of survivors performed: Yes.
- Clinical signs including body weight: Observations included: changes in skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous systems, somatomotor activity, and behavioral pattern. Particular attention was given to potential neurologic endpoints such as tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
- Other examinations performed: All test animals were subjected to gross necropsy.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occured during the study period of 14 days. For details see Table 1 under "Any other information on results incl. tables".

Clinical signs:

other: No clinical signs of toxicity were observed up to the end of the 14-day observation period.

Gross pathology:

Necropsy and histopathological examination revealed no substance-related findings.

Any other information on results incl. tables

Table 1. Acute oral toxicity 

Dose [mg/kg bw]	Mortality	Clinical signs
	N*	N*
Females
2000	0/6	0/6

*N= Number of animals/ number of animals used

Applicant's summary and conclusion

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008.

Conclusions:

The LD50 of the test substance is > 2000 mg/kg bw after single oral administration to Wistar rats.