Registration Dossier

Administrative data

Description of key information

Acute toxicity: oral: 300 < Rat LD50 (female) < 2000 mg/kg bw (OECD 420, GLP, K, rel. 1)
Acute toxicity: dermal: Rat LD50 > 2000 mg/kg bw (OECD 402, GLP, K, rel. 1)

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:
From January 15 to February 10, 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Version / remarks:
Adopted 8th February 2002
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. certificate)
Remarks:
UK GLP Compliance Programme (inspected on August 19, 2008/ signed on March 04, 2009)
Test type:
fixed dose procedure
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK Limited, Bicester, Oxon, UK
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 187-219 g
- Fasting period before study: Animals were fasted for overnight period before test item administration and for approximately 3-4 h after dosing.
- Housing: Animals were housed in groups of up to four in suspended solid-floor polypropylene cages furnished with woodflakes.
- Diet: Food (2014 Teklad Global Rodent diet supplied by Harlan Teklad, Bicester, Oxon, UK), ad libitum
- Water: Mains drinking water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 19-25 °C
- Humidity: 30-70 %
- Air changes: 15 changes / h
- Photoperiod: 12 h dark / 12 h light

IN-LIFE DATES: From: January 15, 2009 To: February 10, 2009
Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 30 and 200 mg/mL
- Justification for choice of vehicle: Test material was not dissolved/suspended in distilled water, therefore arachis oil was selected as vehicle.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION:
- Test material was freshly prepared, as required, as a suspension in arachis oil BP.
Doses:
- Sighting study: 300 and 2000 mg/kg bw
- Main study: 300 mg/kg bw
No. of animals per sex per dose:
- Sighting study: 1 female/dose
- Main study: 5 females/dose (One animal from sighting study and 4 additional animals)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Clinical observations were made 0.5, 1, 2 and 4 h after dosing and then daily for 14 days. Morbidity and mortality checks were made twice daily. Body weight of each animal was recorded on Day 0 (the day of dosing) and on Days 7 and 14 or at death.
- Necropsy of survivors performed: Yes; on completion of the observation period, animals were killed by cervical dislocation and subjected to gross necropsy.
Statistics:
None
Preliminary study:
- There were no deaths or clinical signs of toxicity at a dose level of 300 mg/kg bw.
- Animal treated at a dose level of 2000 mg/kg bw was killed in extremis approximately 2 h after dosing.
- Signs of systemic toxicity noted in animal treated with 2000 mg/kg bw were hunched posture, ataxia, lethargy, pilo-erection, decreased respiratory rate, laboured respiration and hypothermia. The animal was comatose approximately 2 h after dosing.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
>= 300 - < 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: One animal was found dead at 2000 mg/kg bw
Mortality:
- No mortality was observed at a dose level of 300 mg/kg bw.
Clinical signs:
- No signs of systemic toxicity were noted during the observation period at 300 mg/kg bw.
Body weight:
- All animals showed expected gains in bodyweight over the 14 day study period at 300 mg/kg bw.
Gross pathology:
- No abnormalities were noted at necropsy.
Other findings:
None

None

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
Under the test conditions, the oral LD50 for the registered substance is 300 < Oral LD50 ≤2000 mg/kg bw in female rats, therefore the test material is classified as ‘Category 4’ according to the annex VI of the Regulation EC No. 1272/2008 (CLP) and to the GHS. Based on narcotic effects observed at the dose level of 2000 mg/kg bw, the test material is classified as a specific target organ toxicant after single exposure, H336: May cause drowsiness or dizziness according to the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.
Executive summary:

In an acute oral toxicity study performed according to OECD Guideline No. 420 and in compliance with GLP, female Wistar (HsdRccHan®™:WIST™) rats were administered a single oral dose of test material by gavage.

Following a sighting study using one animal at a dose level of 300 and 2000 mg/kg bw, additional four animals were administered a single oral dose of test item at 300 mg/kg bw (main study). Animals were then observed for mortality, clinical signs and bodyweight development for 14 days and at the end of the study the surviving animals were subjected to macroscopic examination.

The animal treated at a dose level of 2000 mg/kg bw was killed in extremis approximately two hours after dosing. Signs of systemic toxicity noted in animal treated with 2000 mg/kg bw were hunched posture, ataxia, lethargy, pilo-erection, decreased respiratory rate, laboured respiration and hypothermia. Animal was comatose approximately two hours after dosing.

No mortality or clinical signs were observed at 300 mg/kg bw. All animals showed expected gains in bodyweight over the 14 day study period. No abnormalities were noted at necropsy.

300 < Rat Oral LD50 (females) ≤2000 mg/kg bw

Under the test conditions, the test material is classified as ‘Category 4’ according to the annex VI of the Regulation EC No. 1272/2008 (CLP) and to the GHS. Based on narcotic effects observed at the dose level of 2000 mg/kg bw, the test material is classified as a specific target organ toxicant after single exposure, H336: May cause drowsiness or dizziness according to the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

This study is considered as acceptable and satisfies the requirement for acute oral toxicity endpoint.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
300 mg/kg bw
Quality of whole database:
The key study is GLP compliant and of high quality (Klimisch score = 1)

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
Not required for substances at the REACH Annex VII tonnage level.

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
Study period:
From July 15 to 29, 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
Adopted 24 February 1987
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. certificate)
Remarks:
UK GLP Compliance Programme (inspected on August 19, 2008/ signed on March 04, 2009)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK Limited, Bicester, Oxon, UK.
- Age at study initiation: 8-12 weeks
- Weight at study initiation: At least 200 g
- Housing: Animals were housed individually during the 24 h exposure period and in groups of five by sex for the remainder of the study in suspended solid-floor polypropylene cages furnished with woodflakes
- Diet: Food (2014 Teklad Global Rodent diet supplied by, Harlan Teklad, Bicester, Oxon, UK), ad libitum
- Water: Mains drinking water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 19-25 °C
- Humidity: 30-70 %
- Air changes: 15 changes/h
- Photoperiod: 12 h dark / 12 h light

IN-LIFE DATES: From: July 15, 2009 To: July 29, 2009
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: Back and flank area
- % coverage: Approximately 10 % of the total body surface area.
- Type of wrap if used: A piece of surgical gauze was placed over the treatment area and semi-occluded with a piece of self-adhesive bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): After the 24 h contact period, the bandage was carefully removed and the treated skin and surrounding hair wiped with cotton wool moistened with distilled water to remove any residual test material.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- Constant volume or concentration used: Yes; 1.78 mL/kg bw
- For solids, paste formed: Yes; the test material was melted in a warming bath set at 60 °C and allowed to cool prior to dosing.
Duration of exposure:
24 h
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed for mortality or clinical signs of toxicity at 0.5, 1, 2 and 4 h after dosing and subsequently once daily for 14 days. Individual bodyweights were recorded prior to application of the test material on Day 0 and on Days 7 and 14.
- Necropsy of survivors performed: Yes; at the end of the study animals were killed by cervical dislocation and subjected to gross necropsy.

Statistics:
None
Preliminary study:
Not applicable
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality was observed.
Mortality:
- No mortality was observed.
Clinical signs:
- No signs of dermal irritation and systemic toxicity were observed.
Body weight:
- Animals showed expected gains in bodyweight over the study period, except for two females which showed no gain in bodyweight and one female which showed bodyweight loss during the first week but expected gain in bodyweight during the second week.
Gross pathology:
- No abnormalities were noted at necropsy.
Other findings:
None

Table 7.2.3/1: Individual bodyweights and bodyweight changes

Dose level

(mg/kg bw)

Animal number

and sex

Body weight (g)

at Day

Body weight gain (g)

during week

0

7

14

1

2

2000

1-0 Male

312

314

332

2

18

1-1 Male

312

322

342

10

20

1-2 Male

319

325

342

6

17

1-3 Male

316

336

343

20

7

1-4 Male

312

316

333

4

17

2-0 Female

200

196

212

-4

16

2-1 Female

201

205

217

4

12

2-2 Female

202

202

212

0

10

2-3 Female

201

207

218

6

11

2-4 Female

208

208

219

0

11

Interpretation of results:
GHS criteria not met
Conclusions:
Under the test conditions, the dermal LD50 for the registered substance is higher than 2000 mg/kg bw in rats therefore it is not classified according to the Annex VI of the Regulation EC No. 1272/2008 (CLP) and to the GHS.
Executive summary:

In an acute dermal toxicity study (limit test) performed according to OECD Guideline No. 402 and in compliance with GLP, a group of Wistar (HsdRccHan®™:WIST®™) rats (5/sex) was given a single dermal application of the undiluted test material at 2000 mg/kg bw to intact skin of the back and flank area at the dose volume of 1.78 mL/kg bw. Test sites were covered with a semi-occlusive dressing for 24 h. Animals were observed for mortality, clinical signs and bodyweights for 14 days and at the end of the study the surviving animals were sacrificed for macroscopic examination. Skin irritation was assessed and scored according to the Draize scale at 24 h after removal of the dressings and then daily for 14 days.

 

No mortality, dermal irritation or systemic toxicity were observed. Animals showed expected gains in bodyweight over the 14 day study period, except for two females which showed no gain in bodyweight and one female which showed bodyweight loss during the first week but expected gain in bodyweight during the second week. No abnormalities were noted at necropsy.

 

Dermal LD50 Combined > 2000 mg/kg bw

 

Under the test conditions, the test material is not classified according to the annex VI of the Regulation EC No. 1272/2008 (CLP) and to the GHS.

 

This study is considered as acceptable and satisfies the requirement for acute dermal toxicity endpoint.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The key study is GLP compliant and of high quality (Klimisch score = 1)

Additional information

Acute toxicity: oral:

A key study was identified (Harlan, 2009, rel. 1). This acute oral toxicity study was performed according to the OECD test guideline No. 420 and in compliance with GLP.

Following a sighting study using one animal at a dose level of 300 and 2000 mg/kg bw, additional four animals were administered a single oral dose of test item at 300 mg/kg bw (main study).

The animal treated at a dose level of 2000 mg/kg bw was killed in extremis approximately two hours after dosing. Signs of systemic toxicity noted in animal treated with 2000 mg/kg bw were hunched posture, ataxia, lethargy, pilo-erection, decreased respiratory rate, laboured respiration and hypothermia. Animal was comatose approximately two hours after dosing.

No mortality or clinical signs were observed at 300 mg/kg bw. All animals showed expected gains in bodyweight over the 14 day study period. No abnormalities were noted at necropsy.

300 < Rat Oral LD50 (females) ≤2000 mg/kg bw

Acute toxicity: dermal:

A key study was identified (Harlan, 2009, rel. 1). This acute dermal toxicity study was performed according to the OECD test guideline No. 402 and in compliance with GLP.

Rats (5/sex) was given a single dermal application of the undiluted test material at 2000 mg/kg bw to intact skin of the back and flank area.

No mortality, dermal irritation or systemic toxicity were observed. Animals showed expected gains in bodyweight over the 14 day study period, except for two females which showed no gain in bodyweight and one female which showed bodyweight loss during the first week but expected gain in bodyweight during the second week. No abnormalities were noted at necropsy.

 

Dermal LD50 Combined > 2000 mg/kg bw

Justification for classification or non-classification

Harmonized classification:

The substance has no harmonized classification according to the Regulation (EC) No. 1272/2008.

Self classification:

Acute toxicity (Oral):

Based on the available information, the substance is classified as H302: Harmful if swallowed (Category 4) according to the Regulation (EC) No. 1272/2008 and to the GHS as the LD50 is between 300 and 2000 mg/kg bw.

Acute toxicity (Dermal):

Based on the available information, the substance is:

- not classified according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) as the dermal LD50 is higher than 2000 mg/kg bw

- not classified according to the GHS since there is no reliable evidence that indicates the LD50 to be in the range of Category 5 values (GHS criteria not met)

Acute toxicity (Inhalation):

No information was available. Not required for substances at the REACH Annex VII tonnage level.

Specific target organ toxicity: single exposure (Oral):

The classification criteria according to the Annex VI of the Regulation (EC) No. 1272/2008 and to the GHS as specific target organ toxicant (STOT) – single exposure, oral Category 3 (H336: May cause droziness or dizziness) are met since narcotic effects, such as lethargy and ataxia were observed following dosing at 2000 mg/kg bw in the acute oral toxicity study and following dosing at 600 mg/kg bw/day in the repeated dose toxicity study.

No non-lethal significant and/or severe toxic effects were observed at the guidance value (oral) for a Category 1 classification (C≤ 300 mg/kg bw) and at the guidance value (oral) for a Category 2 classification (2000 mg/kg bw≥C > 300 mg/kg bw).

Specific target organ toxicity: single exposure (Dermal):

The classification criteria according to the Annex VI of the Regulation (EC) No 1272/2008 and to the GHS as specific target organ toxicant (STOT) – single exposure, dermal are not met since no reversible or irreversible adverse health effects were observed immediately or delayed after exposure and no effects were observed at the guidance value (dermal) for a Category 1 classification (C≤ 1000 mg/kg bw) and at the guidance value (dermal) for a Category 2 classification (2000 mg/kg bw≥C > 1000 mg/kg bw). No classification is required.

The criteria for Transient Organ effects (STOT-SE Category 3) according to Annex VI of the Regulation (EC) No. 1272/2008 and to the GHS are not met since narcotic effects were not observed in the acute dermal toxicity study.

Specific target organ toxicity: single exposure (Inhalation):

No information was available. Not required for substances at the REACH Annex VII tonnage level.