Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Compliance

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:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
Species/strain: healthy rats, WISTAR rats Crl: WI(Han) (full barrier)
Source: Charles River, 97633 Sulzfeld, Germany
Sex: female, non-pregnant, nulliparous
Number of animals:3 per step
Age
at the beginning of the study: 8 - 12 weeks old
Body weight
on the day of administration: Step 1 / animals no. 1 - 3: 177 – 187 g
Step 2 / animals no. 4 - 6: 142 – 162 g
The animals were derived from a controlled full-barrier maintained breeding system (SPF). According to Art. 9.2, No. 7 of the German Act on Animal Welfare the animals were bred for experimental purposes.

Environmental Conditions:
Full barrier in an air-conditioned room
Temperature: 22 ± 3 °C
Relative humidity: 55± 10%
Artificial light, sequence being 12 hours light, 12 hours dark
Air change: 10 x / hour

Free access to Altromin 1324 maintenance diet for rats and mice (lot no. 0815)
Free access to tap water, sulphur acidified to a pH value of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
The animals were kept in groups in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding (lot no. 110811)
Certificates of food, water and bedding are filed at BSL BIOSERVICE
Adequate acclimatisation period (at least five days) under laboratory conditions

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
Dose Administration:
The test item was administered at a single dose by gavage using a feeding tube.
The test item was administered at a dose volume of 10 mL/kg body weight.

Preparation of Dose Formulation:
For all animals of both steps, 2 g of the test item were dissolved in the vehicle to gain a final volume of 10 mL and to achieve a dose of 2000 mg/kg body weight at a dose volume of 10 mL/kg body weight.
Doses:
The starting dose was selected to be 2000 mg/kg body weight. No compound-related mortality was recorded for any animal of step 1 or 2. Based on these results and according to the acute toxic class method regime no further testing was required.
No. of animals per sex per dose:
3 animals per step (2 steps were used)
Control animals:
no
Details on study design:
Preparation of the Animals:
The animals were marked for individual identification by tail painting.
Prior to the administration a detailed clinical observation was made of all animals.
Prior to the administration food was withheld from the test animals for 16 to 19 hours (access to water was permitted). Following the period of fasting the animals were weighed and the test item was administered. Food was provided again approximately 4 hours post dosing.

Observation Period:
All animals were observed for 14 days after dosing for general clinical signs, morbidity and mortality.

Weight Assessment:
The animals were weighed on day 1 (prior to the administration) and on days 8 and 15.

Clinical Examination:
A careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention given during the first 4 hours post-dose). As soon as symptoms were noticed they were recorded. Thereafter, the animals were observed for clinical signs once daily until the end of the observation period.

Pathology:
At the end of the observation period the animals were sacrificed with an overdosage of pentobarbital injected intraperitoneally (Narcoren®, Merial; lot no.: 215061; expiry date: 06/2014) at a dosage of approximately 8 mL/kg bw.
All animals were subjected to gross necropsy. All gross pathological changes were recorded.

Evaluation of Results:
Results were interpreted according to OECD Guideline 423, Annex 2 (see also flow charts in the appendix of the study plan).
Individual reactions of each animal were recorded at each time of observation.
Toxic response data were recorded by dose level.
Nature, severity and duration of clinical observations were described.
Body weight changes were summarised in tabular form.
Necropsy findings were described.
On the basis of the test results, the test substance may be classified in any of the following classes in conformity with the criteria given in Annex VI to Commission Directive 2001/59/EC :
• Very toxic
Substances and preparations shall be classified as very toxic, and assigned the symbol “T+” and indication of danger “very toxic” in accordance with the criteria specified below:
R28 Very toxic if swallowed
- LD50 oral, rat ≤ 25 mg/kg
- less than 100% survival at 5 mg/kg oral, rat by the fixed dose procedure, or
- high mortality at doses ≤ 25 mg/kg oral, by the acute toxic class method.
• Toxic
Substances and preparations shall be classified as toxic, and assigned the symbol “T” and indication of danger “toxic” in accordance with the criteria specified below. Risk phrases shall be assigned in accordance with the following criteria:
R25 Toxic if swallowed
- LD50 oral, rat 25 < LD50 ≤ 200 mg/kg
- discriminating dose, oral rat 5 mg/kg: 100% survival but evident toxicity, or
- high mortality in the dose range > 25 to ≤ 200 mg/kg oral, rat, by the acute toxic class method.
• Harmful
Substances and preparations shall be classified as harmful, and assigned the symbol “Xn” and indication of danger “harmful” in accordance with the criteria specified below. Risk phrases shall be assigned in accordance with the following criteria:
R22 Harmful if swallowed
- LD50 per oral, rat 200 < LD50 ≤ 2000 mg/kg
- discriminating dose, oral rat, 50 mg/kg: 100% survival but evident toxicity,
- less than 100% survival at 500 mg/kg, rat oral by the fixed dose procedure, or
- high mortality in the dose range > 200 to ≤ 2000 mg/kg oral, rat, by the acute toxic class method.
On the basis of the test results, the following risk phrases may be assigned in conformity with the criteria given in Annex I of Regulation (EC) 1272/2008 :
Category 1: LD50 ≤ 5 mg/kg. DANGER. Skull and crossbones in diamond. Fatal if swallowed.
Category 2: LD50 > 5 mg/kg ≤ 50 mg/kg. DANGER. Skull and crossbones in diamond. Fatal if swallowed.
Category 3: LD50 > 50 mg/kg ≤ 300 mg/kg. DANGER. Skull and crossbones in diamond. Toxic if swallowed.
Category 4: LD50 > 300 mg/kg ≤ 2000 mg/kg. WARNING. Exclamation point in diamond. Harmful if swallowed.
On the basis of the test results, the following risk phrases may be assigned in conformity with the criteria given in GHS - Globally Harmonized System of Classification and Labelling of Chemicals, third revised edition, July 2009:
Category 1: LD50 ≤ 5 mg/kg
DANGER. Skull and crossbones in diamond. Fatal if swallowed.
Category 2: LD50 > 5 mg/kg ≤ 50 mg/kg
DANGER. Skull and crossbones in diamond. Fatal if swallowed.
Category 3: LD50 > 50 mg/kg ≤ 300 mg/kg.
DANGER. Skull and crossbones in diamond. Toxic if swallowed.
Category 4: LD50 > 300 mg/kg ≤ 2000 mg/kg.
WARNING. Exclamation point in diamond. Harmful if swallowed.
Category 5: LD50 > 2000 mg/kg ≤ 5000 mg/kg.
WARNING. No symbol. May be harmful if swallowed.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No Mortality was observed in this study.
Clinical signs:
One of six animals (animal No. 6) showed clinical signs on the day of application, which were slightly reduced spontaneous activity,
slight piloerection, half eyelid-closure.
Body weight:
Throughout the 14-day observation period, the weight gain of the animals was within the normal range of variation for this strain.
Gross pathology:
With the exception of acute injection of blood vessels in the abdominal region, which is due to the euthanasia injection, no specific gross pathological changes were recorded for any animal.

Any other information on results incl. tables

Clinical Signs:

Animal
No. / Sex

Time of
Observation
Post-Dose

Observations

 

Step 1 (2000 mg/kg Body Weight)

1 / female

during the whole observation period

no signs of toxicity

2 / female

during the whole observation period

no signs of toxicity

3 / female

during the whole observation period

no signs of toxicity

Step 2 (2000 mg/kg Body Weight)

4 / female

during the whole observation period

no signs of toxicity

5 / female

during the whole observation period

no signs of toxicity

6 / female

30 min

no signs of toxicity

1 h

slightly reduced spontaneous activity,
slight piloerection, half eyelid-closure

2 h

slightly reduced spontaneous activity,
slight piloerection

3 h, 4 h

slight piloerection

d 2 until the end of the observation period

no signs of toxicity

d = day; day 1 = day of administration, h = hour/s, min = minute/s

Body Weight Development:

Animal No. /
Sex

g
Day 1

g
Day 8

g
Day 15

%
Day 1-15

Step 1 (2000 mg/kg Body Weight)

1 / female

177

195

199

12

2 / female

187

214

216

16

3 / female

182

199

208

14

Step 2 (2000 mg/kg Body Weight)

4 / female

162

179

187

15

5 / female

162

183

198

22

6 / female

142

159

174

23

Pathology:

Animal No./
Sex

Organ

Macroscopic Findings

Step 1 (2000 mg/kg Body Weight)

1 / female

-

nsf

2 / female

-

nsf

3 / female

-

nsf

Step 2 (2000 mg/kg Body Weight)

4 / female

-

nsf

5 / female

-

nsf

6 /female

-

nsf

nsf = no specific findings

LD50 Cut-Off:

Dose
(unit)

Number of
Animals
Investigated

Number of Intercurrent Deaths

LD50 Cut-Off

2000 mg/kg bw

6

0

5000 mg/kg bw

bw = body weight

Applicant's summary and conclusion

Interpretation of results:
relatively harmless
Remarks:
Migrated information Criteria used for interpretation of results: OECD GHS
Conclusions:
Under the conditions of the present study, a single oral application of the test item FAT 40858/A TE to rats at a dose of 2000 mg/kg body weight was associated with signs of toxicity.
The median lethal dose of FAT 40858/A TE after a single oral administration to female rats, observed over a period of 14 days is:
LD50 cut-off (rat): 5000 mg/ kg bw
LD50: >2000 mg/kg bw
In conformity with the criteria given in Annex VI to Commission Directive 2001/59/EC the test item FAT 40858/A TE has no obligatory labelling requirement for toxicity.
According to Annex I of Regulation (EC) 1272/2008 the test item FAT 40858/A TE has no obligatory labelling requirement for toxicity and is not classified.
According to GHS (Globally Harmonized Classification System) the test item FAT 40858/A TE has obligatory labelling requirement for toxicity and is classified into Category 5.
Executive summary:

Two steps, each of three female WISTAR Crl: WI(Han) rats, were treated with the test item by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was dissolved in the vehicle aqua ad injectionem (sterile water) at a concentration of 0.2 g/mL and administered at a dose volume of 10 mL/kg body weight.

All animals used in the study after their arrival at BSL were allowed to acclimatise to the laboratory conditions for at least 5 days. The animals were observed on delivery, on inclusion in the study and before administration for mortality/morbidity and other clinical signs. All animals were examined for clinical signs several times on the day of dosing and once daily until the end of the observation period. Their body weights were recorded on day 1 (prior to the administration) and on days 8 and 15.All animals were necropsied and examined macroscopically.

Results per Step:

Step

Sex/No.

Dose (mg/kg)

Number of Animals

Number of Intercurrent Deaths

1

female/1-3

2000

3

0

2

female/4-6

2000

3

0

All animals survived until the end of the study. Only one of six animals showed signs of toxicity.

The clinical findings in this animal treated with the test item at a dose of 2000 mg/kg bw were reduced spontaneous activity, piloerection and half eyelid-closure. These signs are considered to be common symptoms in acute oral toxicity tests.

Throughout the 14-day observation period, the body weight gain of the test animals was within the normal range of variation for this strain.

At necropsy, no macroscopic findings were observed in any animal of any step.


 

On the basis of the test results given below and in conformity with the criteria given in Annex VI to Commission Directive 2001/59/EC, the substance should be:

not classified

X

 

 

limit test

X

On the basis of the test results given below and in conformity with the criteria given inAnnex I of Regulation (EC) 1272/2008, the substance should be:

not classified

X

On the basis of the test results given below and in conformity with the criteria given inGHS (Globally Harmonized System of Classification and Labelling of Chemicals), the substance should be:

classifiedinto category 5

X

LD50cut-off: 5000 mg/kg bw

LD50:  >2000 mg/kg bw

Species/strain: WISTAR Crl: WI(Han) rats

Number of animals: 3 per step / 2 steps performed

Vehicle: aqua ad injectionem

Method: OECD 423

EC 440/2008, Method B.1 tris
OPPTS 870.1000
OPPTS 870.1100