Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
10 May 2012 to 7 June 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
up-and-down procedure
Limit test:
yes

Test material

Constituent 1
Reference substance name:
944127-07-1
IUPAC Name:
944127-07-1
Test material form:
other: solid (unspecified)
Details on test material:
- Appearance: White Solid
- Storage conditions of the test material: Ambient (18 to 36 °C)

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Age at study initiation: 9 to 11 weeks
- Weight at study initiation: 168.85 to 173.49 g. The body weight variation of animals was minimal and did not exceed ± 20 % of the previously dosed animals.
- Fasting period before study: Yes. Animals were fasted overnight prior to dosing (16 to 18 hours). Access to water was not interrupted and the animals were fed immediately after dosing.
- Housing: Rats were housed individually in standard polysulfone cages (Size: approximately 425 mm long x 266 mm deep x 175 mm high), with stainless steel top grills having facilities for pelleted food and drinking water. Steam sterilised clean corn cob bedding was used and changed along with the cage twice weekly. The rats were provided with huts as environmental enrichment in each cage during the experimental period.
- Diet (e.g. ad libitum): Standard rat and mouse pelleted maintenance diet was provided ad libitum
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in a water filter-cum-purifier was provided to animals ad libitum in polycarbonate bottles with stainless steel sipper tubes. The water in the bottles was replenished once daily and the water bottles were changed once a week.
- Acclimation period: Five to fourteen days under standard laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 24 °C
- Humidity (%): 65 to 67 % (relative)
- Air changes (per hr): 12 to 15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours of light and 12 hours of dark

IN-LIFE DATES: From: 10 May 2012 To: 7 June 2012

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: 0.5 % w/v carboxymethyl cellulose sodium salt in Milli-Q water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200 mg/mL as a suspension.
- Justification for choice of vehicle: 2.0 g of the test material was mixed with Milli-Q water and the volume made up to 10 mL. The test material was not miscible. With the addition of 0.5 % w/v carboxymethyl cellulose sodium salt in Milli-Q water, the test material formed a visibly homogenous suspension.

MAXIMUM DOSE VOLUME APPLIED: The volume administered was 10 mL/kg body weight.

DOSAGE PREPARATION: 2 g of test material was mixed by adding small quantities of 0.5 % w/v carboxymethyl cellulose sodium salt in Milli-Q water and the volume was made up to 10 mL. Homogeneity of the test material in the vehicle was maintained during treatment by constant stirring and mixing with a glass rod. Preparations were made prior (within one hour) to dosing.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The limit value was selected as the starting dose.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 females per dose
Control animals:
no
Details on study design:
- Dosing schedule: Single animals were dosed in sequence. Each subsequent animal was dosed only after assessing the toxicity of the test material to the previously dosed animal.
- Duration of observation period following administration: 14 days following test material administration.
- Frequency of observations and weighing: The animals were observed five times on test day 1 (day of administration), at 30 minutes, 1, 2, 3 and 4 hours after dose administration and once daily during days 2 to 15. Individual body weights of animals were recorded on test day 1 (pre-administration), day 8 (7 days post administration) and day 15 (14 days post administration).
- Necropsy of survivors performed: Yes. All animals were necropsied at the end of the 14-day observation period; animals were sacrificed by exsanguination under isoflurane anaesthesia. Animals that died during the observation period were necropsied on the day of death. External surfaces of the body, all orifices, tissues and organs of the thoracic and abdominal cavities of all animals were examined. The stomach was opened, the contents rinsed / removed, and the mucosal surface was examined for signs of irritation, erosions and ulcers. All necropsy observations were recorded.
Statistics:
Dose progression and stopping criteria was calculated using a dedicated software program (Acute Oral Toxicity (Guideline 425) Statistical Program) provided by the EPA.

Results and discussion

Preliminary study:
The limit test at 2000 mg/kg was initiated with one female rat. The first animal survived and there were no clinical signs of toxicity or mortality; hence four additional female rats were tested sequentially at the same dose.
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
- Step 1: There was no mortality.
- Step 2: There was no mortality.
- Step 3: The rat died on day 2.
- Step 4: The rat died on day 2.
- Step 5: There was no mortality.
Clinical signs:
- Step 1: There were no clinical signs of toxicity.
- Step 2: There were no clinical signs of toxicity.
- Step 3: Clinical signs such as slight salivation and lethargy were observed at 3 & 4 hours post treatment.
- Step 4: Clinical signs such as slight salivation and lethargy were observed at 3 & 4 hours post treatment.
- Step 5: Clinical signs such as slight salivation and lethargy were observed at 3 & 4 hours post treatment and the rat was normal from day 2 onwards.
Body weight:
All surviving rats gained body weight throughout the observation period, and the rats that died had lost bodyweight. See Table 1.
Gross pathology:
There were no abnormalities detected at necropsy.

Any other information on results incl. tables

Table 1: Body Weight and Body Weight Changes

Step

Rat Number

Body weight (g)

Initial

(Day 1)

Day 8

Weight change

(Day 8 -Initial)

Day 15

Weight change

(Day 15 -Initial)

At death

1

Rm871

173.49

190.12

16.63

201.23

27.74

-

2

Rm872

173.35

189.94

16.59

203.42

30.07

-

3

Rm873

173.03

-

-

-

-

171.11

4

Rm874

168.85

-

-

-

-

165.01

5

Rm875

171.75

186.92

15.17

201.11

29.36

 

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study, the acute oral LD50 was estimated to be >2000 mg/kg body weight in female Wistar strain rats and the test material requires no classification in accordance with EU criteria.
Executive summary:

The acute oral toxicity potential of the test material to female Wistar strain rats was assessed in accordance with the standardised guidelines OECD 425 and US EPA OPPTS 870.1100 under GLP conditions.

The study was initiated with a limit test at 2000 mg/kg with one female rat. The test material was suspended in 0.5 %w/v carboxymethyl cellulose sodium salt in in Milli-Q water and administered as a single oral dose via gavage to fasted rats. The first animal survived; hence four additional animals were tested sequentially with the same dose.

Three out of the five animals tested showed clinical sings of toxicity such as lethargy and slight salivation on Day 1. Two rats died on Day 2.

As per the Acute Oral Toxicity Software Program (AOT425 StatPgm), the stopping criterion was met and the surviving rats were observed for 14 days post treatment. All the surviving animals had gained body weight during the 14 day observation period.

Rats that died during the observation period were necropsied on the day of death. All surviving animals were sacrificed as scheduled at the end of the observation period. There were no gross abnormalities detected in any of the rats at necropsy.

Under the conditions of this study, the acute oral LD50 was estimated to be >2000 mg/kg body weight in female Wistar strain rats and the test material requires no classification in accordance with EU criteria.