Reaction mass of trimethyl-3-[(1-oxooctadecyl)amino]propylammonium methyl sulphate, Propane-1,2-diol and Trimethyl-3-[(1-oxohexadecyl)amino]propylammonium methyl sulphate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From February 04 to 20, 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of trimethyl-3-[(1-oxooctadecyl)amino]propylammonium methyl sulphate, Propane-1,2-diol and Trimethyl-3-[(1-oxohexadecyl)amino]propylammonium methyl sulphate

EC Number:

947-471-7

Molecular formula:

C22H47N2O.CH3O4S - C3H8O2 - C24H51N2O.CH3O4S

IUPAC Name:

Reaction mass of trimethyl-3-[(1-oxooctadecyl)amino]propylammonium methyl sulphate, Propane-1,2-diol and Trimethyl-3-[(1-oxohexadecyl)amino]propylammonium methyl sulphate

Test material form:

solid

Remarks:

yellowish-brown solid

Test animals

Species:

rat

Strain:

other: Han:WIST rats

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: TOXI COOP ZRT. Cserkesz u. 90. 1103 Budapest, Hungary
- Females (if applicable) nulliparous and non-pregnant: yes
- Fasting period before study: the day before treatment the animals were fasted
- Age at study initiation: young adult rat, 8 weeks old in first, second and third step
- Weight at study initiation: 156 - 166 g at first step, 162 - 174 g at second step, 156 - 165 g at third step
- Housing: group caging (3 animals/cage)
- Diet (e.g. ad libitum): ssniff® SM R/M-Z+H ad libitum
- Water (e.g. ad libitum): tap water from municipal supply ad libitum
- Acclimation period: 5 days in first step, 6 days in second step and 7 days in third step

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 22 ± 3 °C
- Air changes (per hr): above 10 air exchanges/hour by central air-condition system.
- Photoperiod (hrs dark / hrs light): 12 hours daily, from 6.00 a.m. to 6.00 p.m.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Aqua purificata

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30 mg/mL and 200 mg/mL

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: the starting dose was selected on the basis of the available information about the test item. The LD50 value > 200 – 2.000 mg/kg bw according to safety data sheet.

Doses:

300 and 2000 mg/kg bw

No. of animals per sex per dose:

3 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
- Necropsy of survivors performed: yes/no
- Clinical signs including body weight
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

500 mg/kg bw

Based on:

test mat.

Mortality:

No death occurred at 300 mg/kg bw single oral dose of the test item. All female rats in step 1 and step 2 survived until the end of the 14-day observation period.
All rats dosed at 2000 mg/kg bw SJWZPA died on Day 1. The deaths seemed to be consequences of systemic toxic effect of the test item.

Clinical signs:

other: In group 1 treated with 300 mg/kg bw dose no treatment related symptoms were observed throughout the 14-day post-treatment period. In group 2 treated with 300 mg/kg bw dose no treatment related symptoms were observed throughout the 14-day post-treatment p

Gross pathology:

All animals treated with 300 mg/kg bw dose survived until the scheduled necropsy on Day 15.
All animals treated with 2000 mg/kg bw dose spontaneously died during the study and were necropsied on Day 1.
Moderate hydrometra was found in two animals (No.: 7262, 7264) of group 1 and severe hydrometra was detected in animal No.: 7272 of group 2. Hydrometra is physiological finding and connected to the cycle of the animal. No pathological changes were found related to the effect of the test item during the macroscopic examination of animals treated with 300 mg/kg bw dose.
External necropsy findings as anus contaminated with faeces was found in all animals of group 3 (2000 mg/kg bw). Blood around the nose was observed in two animals (No.: 7259, 7260) of group 3. Internal necropsy findings as stomach full of foamy discharge, blushed mucous membrane in the stomach and punctiform haemorrhages on the cardia were found in all animals of group 3 (2000 mg/kg bw). Organs were partially autolised in two animals (No.: 7259, 7260). The intestinal wall was sanguine and intestines were full of yellow and mucous content in animal No.: 7261.

Applicant's summary and conclusion

Interpretation of results:

other: Category 4 based on criteria in the CLP Regulation (EC 1272/2008).

Conclusions:

LD50 value = 500 mg/kg bw

Executive summary:

The acute oral toxicity of the test item was assessed according to the procedure described in the OECD guideline 423.

The starting dose was selected on the basis of the available information about the test item. The LD50 value > 200 – 2.000 mg/kg bw according to safety data sheet.

The acute toxic class method was carried out involving a stepwise procedure with the use of 300 mg/kg bw as the starting dose in three female rats. No animals died in first step, so further three female rats were treated with the same dose. No animals died in second step, too, so further three female rats were treated with higher (2000 mg/kg bw) dose. All animals died in third step.

Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out in animals died on Day 1, as well as 15th day after the treatment in survivor animals.

Lethality, Clinical symptoms and Body weight:

No death occurred at 300 mg/kg bw single oral dose of the test item. All female rats in step 1 and step 2 survived until the end of the 14-day observation period. All rats dosed at 2000 mg/kg bw SJWZPA died on Day 1.

In first step, no clinical symptoms were observed on the day of the treatment and during the 14-day observation period, the general state and behaviour of experimental animals were normal.

In second step, no clinical symptoms were observed on the day of the treatment and during the 14-day observation period, the general state and behaviour of experimental animals were normal.

In third step, CNS - and emotion symptoms (decreased activity, closed eyes, tremor, vocalisation), disturbance of coordination (abnormal gait), decreased righting reflex, decreased muscular - body - tone and a disturbances of the autonomic functions (diarrhea, piloerection) were observed in animals between treatment day (from second hours) and Day 1.

The body weight development was undisturbed in all surviving animals.

Gross pathology:

All animals treated with 300 mg/kg bw dose survived until the scheduled autopsy on Day 15. All organs of the animals treated with 300 mg/kg bw proved to be free of treatment related gross pathological changes.

All animals treated with 2000 mg/kg bw dose died spontaneously during the study and were necropsied on Day 1. External (anus contaminated with faeces, blood around the nose) and internal necropsy findings (stomach and intestines were full of discharge, blushed mucous membrane in the stomach, haemorrhages on the cardia, sanguine intestinal wall) found in animals of group 3 were connected with test item toxic effect.

Conclusion

The estimated LD50 value is 500 mg/kg bw on basis of LD50 cut off mg/kg bw, because three animals died in 2000 mg/kg bw at third step.

The substance is classified for acute toxicity in category 4 based on criteria in the CLP Regulation (EC 1272/2008).