Registration Dossier

Administrative data

Description of key information

Oral (Read-across from structurally similar substance)

LD50 > 2000 mg/kg bw, female rat, OECD 420, EU Method B.1 bis, Bradshaw (2009).

Dermal (Read-across from structurally similar substance)

LD50 > 2000 mg/kg bw, male/female rat, OECD 402, Zelenák (2013).

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
20 January 2009 and 12 February 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in compliance with GLP and agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of relevant results.
Remarks:
Study conducted on read-across material
Justification for type of information:
A RAAF report will shortly be provided.
Reason / purpose for cross-reference:
other: Read-across target
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)
Remarks:
Date of Signature: 15/10/2007 Date of Inspection: 21/08/2007
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK Limited, Bicester, Oxon, UK.
- Age at study initiation: At the start of the study the animals were eight to twelve weeks of age.
- Weight at study initiation: The bodyweight variation did not exceed ± 20% of the initial/mean bodyweight of any previously dosed animal(s).
- Fasting period before study: Overnight fast immediately before dosing and for approximately three to four hours after dosing.
- Housing:The animals were housed in groups of up to four in suspended solid floor polypropylene cages furnished with woodflakes.
- Diet (e.g. ad libitum):Free access to food (2014 Teklad Global Rodent diet supplied by Harlan Teklad, Blackthorn, Bicester, Oxon, UK) was allowed throughout the study.
- Water (e.g. ad libitum): Free access to mains drinking water.
- Acclimation period: Acclimatisation period of at least five days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): The temperature were set to achieve limits of 19 to 25°C.
- Humidity (%): Relative humidity were set to achieve limits 30 to 70%.
- Air changes (per hr): rate of air exchange was at least fifteen changes per hour.
- Photoperiod (hrs dark / hrs light): lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.


IN-LIFE DATES: From:Day 0 To:Day 14
Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200mg/ml
- Amount of vehicle (if gavage):10ml/kg
- Justification for choice of vehicle:. Arachis oil BP was used because the test material did not dissolve/suspend in distilled water.
- Lot/batch no. (if required): Not available
- Purity: Not available


MAXIMUM DOSE VOLUME APPLIED: 10ml/kg


DOSAGE PREPARATION (if unusual):


CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose:Following a sighting test at a dose level of 2000 mg/kg, an additional four fasted female animals were given a single oral dose of test material, as a suspension in arachis oil BP, at a dose level of 2000 mg/kg bodyweight.
Doses:
A total of five animals were treated at a dose level of 2000 mg/kg in the study.
No. of animals per sex per dose:
Starting dose -1 female rat dose level-2000mg/kg
Additional group - 4 female rats dose level-2000mg/kg
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:Clinical observations were made ½, 1, 2, and 4 hours after dosing and subsequently once daily for fourteen days. Morbidity and mortality checks were made twice daily. Individual bodyweights were recorded on Day 0 (the day of dosing) and on Days 7 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:At the end of the observation period the animals were killed by cervical dislocation. All animals were subjected to gross necropsy. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.
Statistics:
None
Preliminary study:
There were no deaths.
No signs of systemic toxicity were noted.
No abnormalities were noted at necropsy.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
There were no deaths.
Clinical signs:
No signs of systemic toxicity were noted.
Body weight:
Individual bodyweights and bodyweight changes are given in Table 2.
All animals showed expected gains in bodyweight over the observation period.
Gross pathology:
Individual necropsy findings are given in Table 3.
No abnormalities were noted at necropsy.
Other findings:
None

Table1              Individual Clinical Observations and Mortality Data

Dose Level mg/kg

Animal Number and Sex

Effects Noted After Dosing
(Hours)

Effects Noted During Period After Dosing
(Days)

½

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2000

1-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-1

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-2

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-3

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 


Table2              Individual Bodyweights and Bodyweight Changes

Dose Level mg/kg

Animal Number and Sex

Bodyweight (g) at Day

Bodyweight Gain (g) During Week

0

7

14

1

2

2000

1-0 Female

208

219

223

11

4

2-0 Female

171

184

193

13

9

2-1 Female

183

187

194

4

7

2-2 Female

186

199

209

13

10

2-3 Female

184

197

211

13

14

 


Table3              Individual Necropsy Findings

Dose Level
mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

2000

1-0 Female

Killed Day 14

No abnormalities detected

2-0 Female

Killed Day 14

No abnormalities detected

2-1 Female

Killed Day 14

No abnormalities detected

2-2 Female

Killed Day 14

No abnormalities detected

2-3 Female

Killed Day 14

No abnormalities detected


0= No signs of systemic toxicity

Interpretation of results:
GHS criteria not met
Conclusions:
The acute oral median lethal dose (LD50) of the test material in the female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight (Globally Harmonised Classification System - Unclassified).
Executive summary:

Introduction. The study was performed to assess the acute oral toxicity of the test material in the Wistar strain rat. The method was designed to meet the requirements of the following:

§        OECD Guidelines for Testing of Chemicals No 420 “Acute Oral Toxicity - Fixed Dose Method” (adopted)

§        Method B1bisAcute Toxicity (Oral) of Commission Directive 2004/73/EC

Method. Following a sighting test at a dose level of 2000 mg/kg, an additional four fasted female animals were given a single oral dose of test material, as asuspensioninarachis oil BP, at a dose level of 2000 mg/kg bodyweight. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.

Mortality. There were no deaths.

Clinical Observations. There were no signs of systemic toxicity.

Bodyweight. All animals showed expected gains in bodyweight.

Necropsy. No abnormalities were noted at necropsy.

Conclusion. The acute oral median lethal dose (LD50) of the test material in the female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight (Globally Harmonised Classification System-Unclassified).

Endpoint:
acute toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study conducted on read-across material
Justification for type of information:
A RAAF report will shortly be provided.
Reason / purpose for cross-reference:
read-across source
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
The read-across study was performed in accordance with standardised guidelines and under GLP conditions and was assigned a reliability score of 1 in accordance with the criteria for assessing data quality as described in Klimisch et al. (1997). The quality of the database is therefore considered to be high.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
30 January 2013 to 13 February 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in compliance with GLP and agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of relevant results.
Remarks:
Study conducted on read-across material
Justification for type of information:
A RAAF report will shortly be provided.
Reason / purpose for cross-reference:
other: Read-across target
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: CRL:(WI) (SPF)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Age at study initiation: Young adults
- Weight at study initiation: 219 to 248 g
- Housing: Individually in type II polypropylene/polycarbonate cages
- Diet: ad libitum
- Water: municipal tap water ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 30 to 70 % (relative)
- Air changes: 15 to 20 air changes per hour
- Photoperiod: 12 hour light/dark cycle (light from 06:00 to 18:00)

IN-LIFE DATES:
From: 24 January 2013
To: 13 February 2013
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: Back. Animals were shaved 24 hours prior to exposure
- % coverage: Approximately 10 %
- Type of wrap if used: Sterile gauze pads held in place by a patch with an adhesive hypoallergenic plaster and the entire trunk of the animal wrapped in a semi-occlusive plastic wrap.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Water (at body temperature)
- Time after start of exposure: 24 hours

TEST MATERIAL
- For solids, paste formed: Yes; the test material was moistened with water to ensure good contact with the skin
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: Clinical observations were performed on the day of treatment at 1 and 5 hours after application and once each day thereafter. Observations included the skin and fur, eyes and mucous membranes, the respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma. Body weights were recorded on Day 0 (prior to test material administration), and on days 7 and 14.
- Necropsy of survivors performed: Yes. All animals were anaesthetised with RELEASE 300 mg/mL inj. A.U.V and exsanguinated. After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs was observed. All macroscopic changes were recorded.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred during the study.
Clinical signs:
No clinical signs, including local dermal signs, were observed after treatment with the test material or during the 14-day observation period.
Body weight:
There were no effects on bodyweight or bodyweight gain after administration of the test material.
Gross pathology:
There was no evidence of any macroscopic changes at necropsy.

Table 1: Bodyweight and Bodyweight Gain for Males

Animal No.

Bodyweight (g)

Bodyweight Gain (g)

Day 0

Day 7

Day 14

Days 0 to 7

Days 7 to 14

Days 0 to 14

9419

237

292

333

55

41

96

9420

248

292

334

44

42

86

9421

245

287

313

42

26

68

9422

234

281

329

47

48

95

9423

234

284

334

50

50

100

Mean

239.6

287.2

328.6

47.6

41.4

89.0

Standard Deviation

6.5

4.9

9.0

5.1

9.4

12.8

 

Table 2: Bodyweight and Bodyweight Gain for Females

Animal No.

Bodyweight (g)

Bodyweight Gain (g)

Day 0

Day 7

Day 14

Days 0 to 7

Days 7 to 14

Days 0 to 14

9424

230

252

262

22

10

32

9425

227

256

272

29

16

45

9426

243

260

273

17

13

30

9427

220

240

275

20

35

55

9428

219

232

251

13

19

32

Mean

227.8

248.0

266.6

20.2

18.6

38.8

Standard Deviation

9.7

11.7

10.1

6.0

9.8

10.8

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study, the acute dermal median lethal dose (LD50) was determined to be > 2000 mg/kg bw in male and female rats.
Executive summary:

The acute dermal toxicity of the test material was determined in male and female CRL:(WI) rats in accordance with the standardised guideline OECD 402 under GLP conditions.

The study was performed as a limit test. 2000 mg/kg bw was applied to the shaved back of 5 animals of each sex (equating to around 10 % of the surface area of the skin) and covered with a semi-occlusive dressing. After 24 hours, the test site was washed with water and the animals were observed for any reactions over a fourteen day observation period. Bodyweights were recorded at days 0, 7 and 14. At the end of the study, all surviving animals were submitted for necropsy.

There was no mortality throughout the study and none of the animals demonstrated any clinical signs of toxicity. Bodyweights and bodyweight gains were not affected by test material administration. All animals were found to be normal at necropsy.

Under the conditions of this study, the acute dermal median lethal dose (LD50) was determined to be > 2000 mg/kg bw in male and female rats.

Endpoint:
acute toxicity: dermal
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study conducted on read-across material
Justification for type of information:
A RAAF report will shortly be provided.
Reason / purpose for cross-reference:
read-across source
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
The read-across study was performed in accordance with standardised guidelines and under GLP conditions and was assigned a reliability score of 1 in accordance with the criteria for assessing data quality as described in Klimisch et al. (1997). The quality of the database is therefore considered to be high.

Additional information

Oral

The key study (Bradshaw 2009) was performed on the read-across substance to assess the acute oral toxicity of the test material in the Wistar strain rat. The method was designed to meet the requirements of OECD 420 and EU Method B.1 bis. The study was performed under GLP conditions and was assigned a reliability score of 1 in accordance with the principles for assessing data quality as defined in Klimisch et al. (1997).

Following a sighting test at a dose level of 2000 mg/kg, an additional four fasted female animals were given a single oral dose of test material, as a suspension in arachis oil BP, at a dose level of 2000 mg/kg bodyweight. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.

There were no deaths and no signs of systemic toxicity. All animals showed expected gains in bodyweight. No abnormalities were noted at necropsy.

The acute oral median lethal dose (LD50) of the test material in the female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight.

In a second study, the acute oral toxicity of the test material was investigated using a single oral administration of 2000 mg/kg in male and female ICR mice of age six weeks. A control group was dosed with vehicle only. The animals were observed for 14 days following administration.

No deaths were observed in male or female mice at the dose level of 2000 mg/kg and so the lethal dose level was estimated to exceed 2000 mg/kg. No abnormalities were observed in any animals in the observations of external appearance, nutritional status and behaviour. There was no significant difference between the body weight development of the treated and control groups and there were no abnormalities in any animals in the macroscopic observations performed.

Under the conditions of this study the oral LD50 value in ICR mice was established to exceed 2000 mg/kg body weight.

Inhalation

In accordance with column 2, point 8.5.2 of Annex VIII of the Regulation (EC) No. 1907/2006 (REACH), testing via the inhalatory route is appropriate if it is a likely route of exposure to humans. As the acute toxicity of the test material has been addressed by the two most likely routes of exposure (oral and dermal), testing for this endpoint is therefore omitted.

 

Dermal

In the key study (Zelenák 2013) the acute dermal toxicity of the read-across material was determined in male and female CRL:(WI) rats in accordance with the standardised guideline OECD 402 under GLP conditions. The study was therefore assigned a reliability score of 1 in accordance with the principles for assessing data quality as defined in Klimisch et al. (1997).

The study was performed as a limit test. 2000 mg/kg bw was applied to the shaved back of 5 animals of each sex (equating to around 10 % of the surface area of the skin) and covered with a semi-occlusive dressing. After 24 hours, the test site was washed with water and the animals were observed for any reactions over a fourteen day observation period. Bodyweights were recorded at days 0, 7 and 14. At the end of the study, all surviving animals were submitted for necropsy.

There was no mortality throughout the study and none of the animals demonstrated any clinical signs of toxicity. Bodyweights and bodyweight gains were not affected by test material administration. All animals were found to be normal at necropsy.

Under the conditions of this study, the acute dermal median lethal dose (LD50) was determined to be > 2000 mg/kg bw in male and female rats.

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No. 1272/2008, the substance does not require classification with respect to acute toxicity via the oral or dermal route.