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Administrative data

Description of key information

LD50, oral rat: >2000 mg/kg bw
LC50, inhal rat: > 0.649 mg/L (active ingredient)
LD50, dermal rat: >2000 mg/kg bw

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
March 05, 1997 through March 26, 1997
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: HanIbm: WIST
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd. Wölferstrasse 4, 4414 Füllinsdorf / Switzerland
- Age at study initiation: Males: 8 weeks; Females: 10 weeks
- Fasting period before study: 16 hours
- Weight at study initiation: Males: 184 - 195 g; Females: 171 - 187 g
- Housing: Groups of five in Makrolon type -4 cages with standard softwood bedding.
- Diet (e.g. ad libitum): Pelleted standard Kliba 343, Batch nos 81/96 and 84/97 rat maintenance diet available ad libitum.
- Water (e.g. ad libitum): Community tap water from Itengen, available ad libitum
- Acclimation period: One week under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3 degrees centigrade
- Humidity (%): 40 - 70%
- Air changes (per hr):10 - 15 per hour
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 2000 mg/kg bw
- Amount of vehicle (if gavage): 10 ml





Doses:
2000 mg/kg administered via gavage as 10 ml/kg in PEG 400
No. of animals per sex per dose:
5 males, 5 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Mortality and viablity observations were four times during test day 1 and once during days 2 - 15. Body weights were recorded on test day 1 (pre-administration), 8 and 15. Each animal was examined for changes in appearance and behaviour four times during day 1, and once daily during days 2 - 15.
- Necropsy of survivors performed: yes
Statistics:
The LOGIT-Model could not be used as no deaths occurred.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
No deaths occurred during the study.
Clinical signs:
other: No clinical signs of toxicity were observed during the study period
Gross pathology:
No macroscopic organ findings were observed at necropsy.

Table 2. Body weights (g) of all test animals during the test period.

Group/Sex

Animal

Day 1

Day 8

Day 15

Group 1/ males (2000 mg/kg)

1

187.1

249.2

272.5

 

2

194.8

250.5

284.9

 

3

188.9

254.3

296.7

 

4

183.6

248.2

280.8

 

5

192.9

255.1

284.7

 

Mean

189.4

251.5

283.9

 

St. Dev.

4.5

3.1

8.7

 

N

5

5

5

 

 

 

 

 

Group 2/females

(2000 mg/kg)

6

186.0

211.0

219.8

 

7

170.5

189.2

207.1

 

8

182.0

201.2

212.5

 

9

186.8

206.3

224.7

 

10

179.4

199.8

217.7

 

Mean

180.9

201.5

216.4

 

St. Dev.

6.6

8.2

6.8

 

N

5

5

5

Conclusions:
The mean lethal dose of CGF-C-1607 after single oral administration to rats of both sexes, observed over a period of 14 days, could not be estimated, because no death occurred: LD50 > 2000 mg/kg
Executive summary:

A group of five male and five female Hanlbm:WIST (SPF) rats was treated with CGF-C-1607 at 2000 mg/kg body weight by oral gavage. The test article was suspended in vehicle (PEG 400) at a concentration of 0.2 g/ml and administered at a volume of 10 ml/kg body weight. Four times during day 1 and once daily during days 2-15 the animals were examined for clinical signs. Mortality/ viability were recorded together with clinical signs at the same time intervals. Body weights were recorded on day 1 before administration and on days 8 and 15. All animals were necropsied and examined macroscopically. No deaths occurred during the study. No clinical signs of toxicity were observed during the observation period. The body weight of the animals was within the range of physiological variability known for rats of this strain and age. No macroscopic organ findings were observed at necropsy.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP guideline study, but restricted for use for purposes of classification and labeling given the air concentration tested relative to the active ingredient.
Qualifier:
according to guideline
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Deviations:
yes
Remarks:
, restricted for use for purposes of classification and labeling given the air concentration tested relative to the active ingredient.
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Species:
rat
Strain:
other: HanRCC: WIST (SPF)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: RCC Ltd; Wölferstrasse 4; 4414 Füllinsdorf; Switzerland
- Age at study initiation: 9 weeks (males); 10 weeks (females)
- Weight at study initiation: 253.4 g - 262.3 g (males); 210.4 g - 220.7 g (females)
- Fasting period before study: no
- Housing: Makrolon Type-IV wire cages with softwood bedding
- Diet (e.g. ad libitum): ad libitum except during 4-hr exposure period
- Water (e.g. ad libitum): ad libitum except during 4-hr exposure period
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20
- Humidity (%): 33 to 70
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: 04-Apr-2007 (day of exposure) To: 18-Apr-2007 (day of necropsy)
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
air
Remarks:
compressed, filtered
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: glass nebuliser (Lomir Biomedical, Inc.)
- Exposure chamber volume: not applicable
- Method of holding animals in test chamber: restraint tubes
- Source and rate of air: compressed, filtered; 18 liters per minute (equated to aerosol flow rate of 1 L/min/animal port)
- Method of particle size determination: Mercer 7-stage cascade impactor (Model 02-130, In-Tox Products, Inc.)
- Temperature, humidity, pressure in air chamber: >22 °C

TEST ATMOSPHERE
- Brief description of analytical method used: collection of aerosol on Millipore HVLP, polyvinylidinediflouride, 0.45 µm pore size filters in 47 mm stainless sampling cassettes. Analysis by gravimetric and HPLC.

- Samples taken from breathing zone: yes

TEST ATMOSPHERE
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): 2.05 µm / 2.66 to 2.10 µm / 2.62
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
6.492 mg/L (SD +- 0.237 mg/L, n=4) based on the analytical measured concentrations of FAT 70'884 consisting of 10% bemotrizinol in phenethyl benzoate. This equates to approximately 0.649 mg/L of bemotrizinol.
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Daily observation, with weighing on days 4, 8, and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, histopathology (macroscopic)
Statistics:
None, given lack of findings.
Sex:
male/female
Dose descriptor:
LC0
Effect level:
> 0.649 mg/L air (analytical)
Based on:
act. ingr.
Remarks:
Bemotrizinol
Exp. duration:
4 h
Sex:
male/female
Dose descriptor:
other: LOAEC
Effect level:
0.649 mg/L air (analytical)
Based on:
act. ingr.
Exp. duration:
4 h
Mortality:
None
Clinical signs:
other: Because there was no vehicle control group and because there are no inhalation studies on the vehicle used, the clinical signs presented here cannot be attributed to the active ingredient. All animals: salivation (slight to moderate in degree), tachypnea
Body weight:
There were no adverse effects on body weight gain of male animals and three of five female animals (male nos. 1 to 5, and female nos. 6, 8 and 9) during the 15-day observation period. Marginal body weight loss or retardation in body weight gain were occasionally seen in the other two female animals, i.e. female no. 7 (+0.4% weight gain from test day 1 to 4 and –0.4% weight loss from test day 8 to 15) and female no. 10 (+0.8% weight gain from test day 4 to 8). In view of the concomitant clinical signs noted in all animals, a relationship of these minor effects on female body weight to the treatment with the test item could not be entirely discounted, although slight physical stress, e.g. during restraint in the exposure tubes, may have contributed to these effects.
Gross pathology:
No macroscopic findings.

Body Weights (grams)

Group

Animal
No.

Sex

Test Day 1
(Treatment)

Test Day 4

Test Day 8

Test Day 15

1

(6.492 mg/L)

1

M

255.2

264.9

276.0

306.7

2

M

262.3

268.8

290.5

325.0

3

M

253.4

259.6

266.8

290.0

4

M

257.7

266.8

280.9

311.9

Mean

257.4

265.6

278.9

308.3

+S.D.

3.4

3.7

8.6

12.6

N

5

5

5

5

 *Group 1 (10% active ingredient): 0.649 mg/L (active ingredient)

Group

Animal
No.

Sex

Test Day 1
(Treatment)

Test Day 4

Test Day 8

Test Day 15

1

(6.492 mg/L)

6

F

220.7

231.5

235.7

242.4

7

F

213.2

214.1

216.9

216.0

8

F

210.4

214.2

218.7

225.6

9

F

217.9

224.0

226.6

239.6

Mean

216.2

221.1

224.3

231.4

+S.D.

4.3

7.3

7.4

10.7

N

5

5

5

5

*Group 1 (10% active ingredient): 0.649 mg/L (active ingredient)

Conclusions:
The LC50 of FAT 70’884 [undiluted formulation comprising approximately 10% FAT 70’884 in X-Tend (phenethyl benzoate)] obtained in this study was estimated to be >6.492 mg/L air (chemically determined mean aerosol concentration), which equates to >0.649 mg/L for the active ingredient. There was no indication of relevant sex-related differences in toxicity of the test item.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: HanIbm: WIST
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd. Wolferstrasse 4, 4414 Fullinsdorf / Switzerland
- Age at study initiation: Males: 8 weeks; Females: 11 weeks
- Fasting period before study: 16 hours
- Weight at study initiation: Males: 231.7 - 256.9 g; Females: 190.4 - 219.8 g
- Housing: Groups of five in Makrolon type -4 cages with standard softwood bedding.
- Diet (e.g. ad libitum): Pelleted standard Kliba 343, Batch nos 81/96 and 84/97 rat maintenance diet available ad libitum.
- Water (e.g. ad libitum): Community tap water from Itengen, available ad libitum
- Acclimation period: One week under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3 degrees centigrade
- Humidity (%): 40 - 70%
- Air changes (per hr):10 - 15 per hour
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
semiocclusive
Vehicle:
polyethylene glycol
Remarks:
(PEG 400)
Details on dermal exposure:
TEST SITE
- Area of exposure: backs
- % coverage: 10
- Type of wrap if used: elastic adhesive bandage

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes, with lukewarm tap water
- Time after start of exposure: 24 hrs

TEST SOLUTION
- Amount(s) applied (volume or weight with unit): 4 ml/kg bw
- Concentration of test material: 0.5 g/ml
- Dose: 2000 mg/kg bw
- For solids, paste formed: no (suspension in PG400)
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Daily, with weighings on days 1 (prior to exposure), 8, and 15
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, histopathology
Statistics:
None, as no deaths ocurred.
Sex:
male/female
Dose descriptor:
LD0
Effect level:
2 000 mg/kg bw
Based on:
test mat.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
None
Clinical signs:
other: None
Gross pathology:
No abnormalities

 

Group / Sex

Animal

Day 1

Day 8

Day 15

1 / Males

(2000 mg/kg bw)

1

245.8

271.9

299.9

2

243.4

265.0

299.1

3

256.9

284.0

320.2

4

242.5

272.9

306.1

5

231.7

266.5

300.1

Mean

244.1

272.1

305.1

+S.D.

9.0

7.5

8.9

N

5

5

5

1 / Females

(2000 mg/kg bw)

6

209.4

211.0

220.7

7

206.4

207.5

214.5

8

219.8

225.5

229.0

9

219.5

227.5

237.8

10

190.4

204.4

212.6

Mean

209.1

215.2

222.9

+S.D.

12.0

10.6

10.5

N

5

5

5

 

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
No mortality, clinical signs of toxicity, or macroscopic abnormalities were observed at the dermally applied limit dose of 2000 mg/kg bw.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Additional information

Bemotrizinol is not expected to cause acute toxicity via oral and dermal routes. In rats, oral LD50s are >2000 mg/kg bw (RCC, 1997), LC50 is > 0.6492 mg/L (RCC, 2008), and dermal LD50s are >2000 mg/kg bw (RCC, 1997). No clinical signs were reported in either oral or dermal studies. Clinical signs due to inhalation of 10% bemotrizinol in phenethyl benzoate include salivation, tachypnea, restlessness, hunched posture and ruffled fur. However, since there was no control group exposed to phenethyl benzoate only and there do not exist inhalation studies with phenethyl benzoate, the clinical signs reported cannot be attributed exclusively to the active ingredient.

Justification for classification or non-classification

Acute oral toxicity

Based on the results of acute oral toxicity testing (LD50 > 2000 mg/kg), Bemotrizinol is not expected to cause any adverse effects and would not be classified under the EU DSD classification criteria or under the EU CLP classification criteria (EU Regulation 1272/2008).

Acute inhalation toxicity

The results obtained from inhalation study with rats of 10% bemotrizinol in phenethyl benzoate indicate the LC50 > 0.6492 mg/L.

Considering the rules of CLP, this would mean that bemotrizinol is still potentially classified as acute toxic cat. 4. However, combining this result with the lack of toxicity in the acute oral and dermal studies, it can be safely concluded that bemotrizinol is not hazardous in case of short-term inhalation.

Acute dermal toxicity

Based on the results of the study (rat dermal LD50 >2000 mg/kg), Bemotrizinol would not be classified under the EU DSD classification criteria or under the EU CLP classification criteria (EU Regulation 1272/2008).