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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2009-10-20 to 2009-11-25
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted 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. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
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

Constituent 1
Chemical structure
Reference substance name:
Trimanganese tetraoxide
EC Number:
215-266-5
EC Name:
Trimanganese tetraoxide
Cas Number:
1317-35-7
Molecular formula:
Mn3O4
IUPAC Name:
trimanganese tetraoxide
Test material form:
solid: particulate/powder
Details on test material:
- Molecular formula of test material: Mn3O4
- Appearance: Brown powder
- Physical state: Solid
- Impurities (identity and concentrations): K2O - 1 ppm, Na2O - 3 ppm, MgO -44 ppm, CaO- 14 ppm, SiO2- 12ppm & S0 0.041%
- Composition of test material, percentage of components: Mn 71.2 %
- Purity test date: 26/09/08
- Lot/batch No.: 08100
- Storage condition of test material: Room temperature in the dark

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK, Ltd. Bicester, Oxon, UK
- Age at study initiation: 8 to 12 weeks
- Weight at study initiation: The bodyweight variation did not exceed ± 20 % of the initial/mean bodyweight of any previously dosed animals.
- Fasting period before study: Overnight fasting prior to dosing, and 3 to 4 hours post dosing.
- Housing: Animals were housed in groups of up to four in suspended solid-floor polypropylene cages furnished with woodflakes.
- Diet: 2014 Teklad Global Rodent diet supplied by Harlan Teklad Blackthorn (Bicester, Oxon, UK) available ad libitum
- Water: Mains tap water available ad libitum
- Acclimation period: A minimum of 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25 °C
- Humidity (%): 30 to 70 %
- Air changes (per hr): A minimum of 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hour cycle

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 30 mg/mL at the 300 mg/kg dose level, 200 mg/mL at the 2000 mg/kg dose level
- Amount of vehicle (if gavage): 10 mL/kg
Doses:
2,000 mg/kg employed in the main test and 300 and 2,000 mg/kg in the sighting test.
No. of animals per sex per dose:
1 animal in each of the sighting dose levels, and 4 animals in the main test.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were weighed on days 0 and 7. Clinical observations were made at 30 minutes, then 1, 2 and 4 hours post dosing and then daily for fourteen days. Morbidity and mortality checks were made twice daily.
- Necropsy of survivors performed: yes, animals were killed by cervical dislocation at the end of the 14 day observation period
- Other examinations performed: clinical signs, body weight and histopathology
Statistics:
Data evaluations included the relationship (if any were noted) between the animal's exposure to the test material and the incidence and severity of all abnormalities including behavioural and clinical observations, gross lesions, bodyweight changes, mortality and any other toxicological effects. If possible the signs of evident toxicity were also identified. Evident toxicity is defined as the toxic effects which are of a severity such that administration at the next highest level could result in mortality.

Using mortality data, an estimate of the acute oral median lethal dose (LD50) of the test material was made.

Results and discussion

Preliminary study:
At both levels of dosing in the sighting study, all animals showed expected bodyweight gains over the observation period. No signs of systemic toxicity were noted, and at necropsy.
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Remarks on result:
other: No signs of toxicity were noted at either 300 or 2000 mg/kg
Mortality:
No mortalities occurred during the study.
Clinical signs:
No signs of toxicity were noted during the study.
Body weight:
All animals exhibited expected bodyweight gains during the course of the study.
Gross pathology:
No macroscopic abnormalities were recorded at necropsy.

Any other information on results incl. tables

 Table 2. Individual Clinical Observations and Mortality Data – 2000 mg/kg

Dose level mg/kg

Animal Number and Sex

Effects noted after dosing

Effects noted during periods after dosing (days)

1/2

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2000

2-0 Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-0 Female

0

0

0

0

*

0

0

0

0

0

0

0

0

0

0

0

0

0

3-1 Female

0

0

0

0

*

0

0

0

0

0

0

0

0

0

0

0

0

0

3-2 Female

0

0

0

0

*

0

0

0

0

0

0

0

0

0

0

0

0

0

3-3 Female

0

0

0

0

*

0

0

0

0

0

0

0

0

0

0

0

0

0

* - Due to a technician error day 1 observation not performed

0 – No sign of systemic toxicity

  

Table 3: Individual Bodyweights and Bodyweight Changes -2000 mg/kg

Dose Level mg/kg

Animal Number and Sex

Bodyweights (g) at Day

Bodyweight Gain (g) During week

0

7

14

1

2

2000

2-0 Female

181

191

192

10

1

3-0 Female

159

171

179

12

8

3-1 Female

163

172

173

9

1

3-2 Female

174

188

193

14

5

3-3 Female

176

204

204

28

0

 

Table 4: Individual Necropsy Findings – 2000 mg/kg

Dose Level mg/kg

Animal Number and Sex

Time of Death

Macroscopic Observation

 

 

 

 

2000

2-0 Female

Killed Day 14

No abnormalities detected

3-0 Female

Killed Day 14

No abnormalities detected

3-1 Female

Killed Day 14

No abnormalities detected

3-2 Female

Killed Day 14

No abnormalities detected

3-3 Female

Killed Day 14

No abnormalities detected

 

 

 

 

 

Applicant's summary and conclusion

Interpretation of results:
study cannot be used for classification
Conclusions:
The acute oral median lethal dose (LD50) of the test material in the female Wistar rat was determined to be greater than 2000 mg/kg bw under the conditions of the test.
Executive summary:

The acute oral toxicity of the test material was investigated in a study which was conducted under GLP conditions and in accordance with the standardised guidelines OECD 420 and EU Method B.1 bis.

The acute oral median lethal dose (LD50) of the test material in the female Wistar rat was determined to be greater than 2000 mg/kg bw under the conditions of the test.