Ethanol, 2,2'-iminobis-, N-tallow alkyl derivs., N-oxides

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21 May - 29 June, 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

17 December 2001

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Version / remarks:

30 May 2008

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

December 2002

Deviations:

no

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:

Ethanol, 2,2'-iminobis-, N-tallow alkyl derivs., N-oxides

EC Number:

263-179-6

EC Name:

Ethanol, 2,2'-iminobis-, N-tallow alkyl derivs., N-oxides

Cas Number:

61791-46-6

Molecular formula:

n.a

IUPAC Name:

Ethanol, 2,2'-iminobis-, N-tallow alkyl derivs., N-oxides

Test material form:

semi-solid (amorphous): gel

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: In-house bred animals, Animal Facility, Bioneeds India Private Limited, Devarahosahally, Sompura Hobli, Nelamangala Taluk, Bangalore Rural District, PIN - 562 111, Karnataka, India
- Females: Yes
- Age at study initiation: young adult, 8 to 12 weeks
- Weight at study initiation: 160 - 180 g
- Housing: up to three animals in standard polypropylene cage with stainless steel mesh top grill.
- Diet: Altromin Maintenance diet for rats and mice 1324 (Altromin Spezialfutter GmbH & Co. KG), ad libitum
- Water: Deep bore-well water passed through reverse osmosis unit, provided in plastic water bottles with stainless steel sipper tubes, ad libitum
- Acclimation period: min. 5 days
- Fasting period before study: Three animals per step will be fasted overnight (16 to 18 hours) prior to dosing.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): 12 -15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Remarks:

0.5% w/v

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30 mg/mL
- Amount of vehicle: 10 mL
- Justification for choice of vehicle: The test item forms uniform suspension in 0.5% w/v Carboxy Methyl Cellulose (CMC) as evidenced by the in-house solubility test. Hence, 0.5% w/v CMC has been selected for the test item formulation preparation.0.5% w/v CMC is universally accepted and routinely used vehicle in oral toxicity studies.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION:
The required quantity of test item was weighed into a beaker and mixed well using a glass rod by adding little volume of vehicle. Then it was transferred into the measuring cylinder. Again a small quantity of vehicle was added, mixed well and transferred into the measuring cylinder. The rinsing procedure was repeated to ensure complete transfer of the test item formulation into the measuring cylinder. Finally, the volume was made up to the required quantity with vehicle to get the desired concentration. Freshly prepared formulation was used for dosing. The homogeneity of the test item formulation was maintained by continuous stirring on a magnetic stirrer during dosing (if required).


CLASS METHOD
- Rationale for the selection of the starting dose: A starting dose of 300 mg/kg body weight has been selected from the fixed dose levels of 5, 50, 300 and 2000 mg/kg body weight as there is no available information on the LD50 of the test item.

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: All the animals were observed for clinical signs of toxicity and mortality at 30 to 40 min, 1 hr (±10 min), 2 hrs (±10 min), 3 hrs (±10 min) and 4 hrs (±10 min) post dosing on Day 1 and at least once daily thereafter for clinical signs of toxicity throughout the experimental period. However, more frequent observations were carried out based on the severity of the clinical signs observed and twice daily for mortality during the experimental period. Individual animal body weight was recorded at receipt, on day 1 before test item administration and on day 8 and 15 during the observation period. If the animal is found dead, then the body weight of dead animal was recorded.
- Necropsy of survivors performed: Yes, at the end of observation period, all surviving animals were sacrificed under carbon dioxide/isoflurane anaesthesia. All animals (including those that die during the test or are removed from the study for animal welfare reasons) were subjected to gross pathology
- Other examinations performed: clinical signs, body weight, histopathology

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat. (total fraction)

Mortality:

No mortality was observed during the study

Clinical signs:

No clinical signs were observed during the study

Body weight:

No abmormal body weight development was observed.

Gross pathology:

No gross pathological changes were observed in any of the animals.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In an in vivo acute oral toxicity study according to OECD guideline 423, an LD50 of above 2000 mgkg/kg bw was determined.

Executive summary:

The test item was evaluated for acute oral toxicity in young adult female Sprague Dawley rats according to OECD guideline 423 (Acute Toxic Class Method, adopted on 17 December 2001).

A starting dose of 300 mg/kg body weight was selected as there was no available information on the LD50 of the test item. A total of 12 females (3 females each for each Step-I, Step-l confirmation, Step-II and Step-ll confirmation) were used for the experiment. All the animals of Step-I and Step-I confirmation were administered with 300 mg/kg body weight of the test item and Step II and Step II confirmation were administered with 2000 mg/kg body weight of the test item by oral route.

All animals were observed for clinical signs of toxicity and mortality at 30 to 40 min, 1 h (±10 mins), 2 h (±10 mins), 3 h (±10 mills) and 4 h (±10 mins) post dosing on day 1. Thereafter, observation was performed once daily for clinical signs of toxicity and twice daily for mortality during the 14 days observation period. Body weight was recorded at receipt, on day 1 before test item administration and on day 8 and 15 during the observation period. At the end of observation period, all surviving animals were sacrificed under carbon dioxide anesthesia and subjected to necropsy and gross pathological examination.

Animals did not reveal any clinical signs of toxicity and mortality. No changes were observed in body weight and percent change in body weight after treatment. All the animals revealed physiologically normal body weight development. No gross pathological changes were observed in any of the animals.

Based on the results of the experiment, it is concluded that the LD50 cut off value for the test item is 2000 mg/kg body weight when administered as a single dose by oral gavage to female Sprague Dawley rats. Accordingly, no classification for acute oral toxicity is warranted.