[No public or meaningful name is available]

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30 August 2016 to 22 September 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Test material form:

liquid

Details on test material:

- Appearance/physical state: Colourless to yellow liquid
- Storage conditions: Room temperature in the dark

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

ANIMAL INFORMATION
- Female Wistar strain rats (RccHan:WIST) were supplied by Envigo RMS (UK) Limited, Oxon, UK.
- On receipt the animals were randomly allocated to cages.
- The females were nulliparous and non-pregnant.
- After an acclimatisation period of at least 5 days the animals were selected at random and given a number unique within the study by indelible ink marking on the tail and a number written on a cage card.
- At the start of the study the animals were 8 to 12 weeks of age.
- Body weight variation did not exceed ± 20 % of the mean body weight at the start of treatment.

ANIMAL CARE AND HUSBANDRY
- Animals were housed in groups of up to four in suspended solid-floor polypropylene cages furnished with wood flakes.
- With the exception of an overnight fast immediately before dosing and for approximately 3 to 4 hours after dosing, free access to mains drinking water and food (2014C Teklad Global Rodent diet supplied by Envigo RMS (UK) Limited, Oxon, UK) was allowed throughout the study.
- The diet, drinking water and bedding were routinely analysed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.
- Temperature was set to achieve limits of 19 to 25 °C.
- Relative humidity was set to achieve limits of 30 to 70 %.
- Rate of air exchange was at least 15 changes per hour.
- Lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness.
- Animals were provided with environmental enrichment items which were condidered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

TEST ITEM PREPARATION AND ANALYSIS
- The test item was used as supplied.
- Specific gravity was determined and used to calculate the appropriate dose volume for the required dose level.

EXPOSURE TO TEST ITEM
- All animals were dosed once only by gavage using a metal cannula attached to a graduated syringe.
- The volume administered to each animal was calculated according to the fasted body weight at the time of dosing.
- Treatment of animals was sequential.
- Sufficient time was allowed between each dose group to confirm the survival of previously dosed animals.
- Clinical observations were made 0.5, 1, 2 and 4 hours after dosing and then daily for 14 days.
- Morbidity and mortality checks were made twice daily (early and late) during normal working days and once daily on weekends and public holidays.
- Individual body weights were recorded on Day 0 (the day of dosing) and on days 7 and 14.
- At the end of the observation period the animals were killed by cervical dislocation.
- All animals were subjected to gross necropsy consisting of an external examination plus opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

DATA EVALUATION
- The test item was evaluated according to Annex 3 of the OECD Guidelines for Testing of Chemicals No 420 “Acute Oral Toxicity – Fixed Dose Method” (adopted 17 December 2001) as shown in the flow chart in Annex 2 (attached).
- Evaluation of data included identification of the number of animals that died during the study (or were killed for humane reasons) plus determination of the nature, severity, onset and duration of toxic effects. If possible, the signs of evident toxicity were described.
- Evident toxicity refers to toxic effects of sufficient severity that administration of the next higher dose level could result in development of severe signs of toxicity and probably mortality.
- Effects on body weights and abnormalities at necropsy were also identified.

Doses:

Single dose of 2000 mg/kg bw

No. of animals per sex per dose:

Five

Control animals:

no

Details on study design:

STUDY DESIGN
- Flow charts summarising the sighting test and main test are provided in Annex 2 and Annex 3 (attached).
- Using available information on the toxicity of the test item, the starting dose was chosen as 2000 mg/kg bw.
- A single animal was treated with 2000 mg/kg bw (specific gravity 0.970; dose volume 2.07 mL/kg).
- In the absence of toxicity, an additional group of four animals was then treated with 2000 mg/kg bw (specific gravity 0.970; dose volume 2.07 mL/kg).
- A total of five animals was therefore treated at the dose level of 2000 mg/kg bw during the study.

Statistics:

- An estimate of the acute oral median lethal dose (LD50) of the test item was made using mortality data obtained during the study.

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 animal deaths took place during the study.

Clinical signs:

other: - No signs of systemic toxicity were noted during the observation period.

Gross pathology:

- Individual necropsy findings are given in Appendix 3 (attached).
- No abnormalities were noted at necropsy.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was found to be > 2000 mg/kg bw.

Executive summary:

GUIDELINE

The study was performed to assess acute oral toxicity of the test item in the Wistar strain rat in compliance with the OECD Guideline for Testing of Chemicals No 420 “Acute Oral Toxicity – Fixed Dose Method” (2001) and Method B.1 bis Acute Toxicity (Oral) of Commission Regulation (EC) No 440/2008.

 

METHODS

Following a sighting test at a dose level of 200 mg/kg, an additional four fasted female animals were given a single oral dose of test item at a dose level of 2000 mg/kg bw. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

 

RESULTS

No animal deaths took place during the study and clinical observations resulted in no reports of systemic toxicity. All animals showed expected gains in body weight and no abnormalities were noted at necropsy.

 

CONCLUSION

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was found to be > 2000 mg/kg bw.