Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 Feb - 28 Mar 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
adopted in 2001
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
adopted in 2008
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
adopted 2002
Qualifier:
according to
Guideline:
other: Food and Agricultural Materials Inspection Centre (FAMIC), 12 Nohsan, Notification No. 8147
Version / remarks:
adopted in 2011
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed

Test animals

Species:
rat
Strain:
Wistar
Remarks:
Crl:WI (Han)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:
Charles River Deutschland, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: approx.
8 or 9 weeks
- Fasting period before study:
animals were fasted overnight and until 3 - 4 h after administration
- Housing:
3 animals per cage in labeled Makrolon cages (MIV type; height 18 cm) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
- Diet:
pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany), ad libitum
- Water: tap water, ad libitum
(analysis were performed)
- Acclimation period:
at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C):
18 - 24
- Humidity (%):
40 - 70
- Air changes (per hr):
at least 10
- Photoperiod (hrs dark / hrs light):
12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Details on oral exposure:
VEHICLE
- Concentration vehicle: 1% aqueous carboxymethyl cellulose
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed at WIL Research Europe and on test substance data supplied by the sponsor.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw
Doses:
2000 mg/kg bw
300 mg/kg bw
No. of animals per sex per dose:
3 females per step (4 steps)
Control animals:
no
Remarks:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15 clinical signs were recorded. The symptoms were graded according to fixed scales and the time of onset, degree and duration was recorded. Mortality/Viability was recorded twice daily. The time of death was recorded as precisely as possible.
- Necropsy of survivors performed: yes
Statistics:
Mean values and standard deviations were calculated for body weights.

Results and discussion

Effect levelsopen allclose all
Sex:
female
Dose descriptor:
LD50 cut-off
Effect level:
2 000 mg/kg bw
Based on:
test mat.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Step 1: At 2000 mg/kg bw, 1 of 3 animals was sacrificed for humane reasons on Day 2.
Step 2: At 2000 mg/kg bw, all 3 animals were found dead on Day 2.
Step 3+4: At 300 mg/kg bw, no mortality occurred during the study period.
Clinical signs:
At 2000 mg/kg bw, lethargy, flat posture, hunched posture, lateral recumbency, uncoordinated movements, piloerection, chromodacryorrhoea, clonic spasms, quick breathing and/or watery discharge from the eye were noted for the animals. The surviving animals had recovered from the symptoms by Day 9. At 300 mg/kg bw, lethargy, hunched posture, uncoordinated movements, piloerection and/or chromodacryorrhoea were noted for the animals on Days 1 and/or 2.
Body weight:
Normal body weight gains were observed in all animals.
Gross pathology:
At 2000 mg/kg bw, abnormalities of the body cavities (thoracic cavity, diaphragm: dark red, hard, nodule 8x6 mm), small intestines (contents: black discolouration), caecum (contents: black, hard) and/or emaciation were found in the animals that died or were sacrificed for humane reasons during the study, at macroscopic post mortem examination. Macroscopic examination of the surviving animals at
termination did not reveal any abnormalities.
At 300 mg/kg bw, abnormalities of the left kidney (enlarged, contents: watery-clear), ureter (left side: dilation) and/or thymus (isolated, reddish focus/foci) were found in two animals, at macroscopic examination. Macroscopic examination of the other animals at termination did not reveal any abnormalities.

Any other information on results incl. tables

Table 1: Absolute body weights

Starting dose (mg/kg bw)

Animal No.

Body weight (g)

Day 1

Day 8

Day 15

2000

1

152

177

198

2

148

171

189

3

135

*

-

Mean ± SD

145 ± 9

174 ± 4

194 ± 6

4

181

**

-

5

168

**

-

6

175

**

-

Mean ± SD

175 ± 7

-

-

300

7

181

209

214

8

197

230

241

9

177

206

210

Mean ± SD

185 ± 11

215 ± 13

222 ± 17

10

162

188

195

11

172

188

212

12

160

189

194

Mean ± SD

165 ± 6

188 ± 1

200 ± 10

* Animal sacrificed for humane reasons on Day 2, bodyweight at death 125 g

** Animals found dead on Day 2, body weights at death 171 g, 157 g and 167 g, respectively

Applicant's summary and conclusion

Interpretation of results:
other: CLP/EU GHS Category 4 (H302) according to Regulation (EC) No 1272/2008
Conclusions:
CLP: Acute Oral 4, H302

An acute oral toxicity study according to OECD Guideline 423 in rats was performed with the test substance and resulted in a LD50 between 300 and 2000 mg/kg bw. Thus the test substance does meet the classification criteria according to Regulation (EC) 1272/2008.