Registration Dossier

Administrative data

Description of key information

Oral:

LD50 of between 500 and 1000 mg/kg.

OECD 423, GLP (Stepan, 2000), Sprague-Dawley rats exposed to 200 mg/kg (only females) and 2000 mg/kg (males and females). Two animals treated with 2000 mg/kg were found dead one or two days after dosing. Necropsy results revealed dark liver and dark kidneys in the 2000 mg/kg group. 

Dermal:

LD50 greater than 2000 mg/kg

OECD 402, GLP (Stepan, 2000), 5 Sprague-Dawley rats exposed to 2000 mg/kg. No observed mortality. In four animals, a slight brown discoloration was observed.

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 October 20, 1999 to November 24, 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
March 1996
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source: Stepan Company, Illinois, USA
- Batch No.of test material: 876 TK
- Date received: 10 May 1999
- Description: white paste

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Freshly prepared as a solution in distilled water
- Final dilution of a dissolved solid: The dose levels were selected using a correction factor based on the purity of the supplied test material, to ensure that the animals received equivalent to 200 or 2000 mg/kg of pure test material

FORM AS APPLIED IN THE TEST: solution
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd, Margate, Kent, UK
- Age at study initiation: eight to twelve weeks old
- Weight at study initiation: males weighed 228 to 236 g, and the females 201 to 236 g
- Fasting period before study: overnight immediately before dosing
- Housing: in groups of three by sex in solid-floor polypropylene cages furnished with wood flakes
- Diet (e.g. ad libitum): ad libitum with the exception of an overnight fast immediately before dosing and for approximately three to four hours after dosing
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25 ºC
- Humidity (%): 30 - 70 %
- Air changes (per hr): The rate of air exchange was approximately fifteen changes per hour
- Photoperiod (hrs dark / hrs light): cycles of twelve hours continuous light and twelve hours darkness.
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 217.4 mg/ml and 21.8 mg/ml
- Amount of vehicle (if gavage): 10 ml

DOSAGE PREPARATION: The test material solutions were prepared using a correction factor based on the purity of the test substance to ensure that the animals received 2000 and 200 mg/kg of the pure test material

MAXIMUM DOSE VOLUME APPLIED: 10 mL

CLASS METHOD
- Rationale for the selection of the starting dose: Using all available information, 2174 mg/kg bodyweight was selected as the starting dose. (This dose level was selected using a correction factor based on the purity of the supplied test material, to ensure that the animals received a dose equivalent to 2000 mg/kg bodyweight of pure test material).

OTHER:
- animals were dosed once only by gavage
Doses:
2174 mg/kg (equivalent to 2000 mg pure test material/kg) and 218 mg/kg (equivalent to 200 mg pure test material/kg)
No. of animals per sex per dose:
2174 mg/kg: a group of 3 female rats.
218 mg/kg: 6 rats, one group of 3 females and one group of 3 males
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Observation: Death or overt signs of toxicity were checked at 1/2, 1, 2, and four hours after dosing and subsequently once daily
- Frequency of observations and weighing: Individual bodyweights were recorded prior to dosing and seven and fourteen days after treatment or at death
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs and body weight.
Sex:
female
Dose descriptor:
LD50
Effect level:
>= 500 - <= 1 000 mg/kg bw
Based on:
act. ingr.
Remarks on result:
other: Pure test substance
Mortality:
Two animals treated with 2174 mg/kg were found dead one or two days after dosing. There were no deaths noted at a dose level of 218 mg/kg.
Clinical signs:
Clinical signs in animals treated with 2174 mg/kg were hunched posture, diarrhoea and increased salivation with incidents of pallor of the extremities, emaciation, lethargy, pilo-erection, decreased respiratory rate, laboured respiration, red/brown staining around snout and tiptoe gait. The surviving female animals treated with 2174 mg/kg recovered five days after dosing. All animals treated with 218 mg/kg appeared normal throughout the study.
Body weight:
All surviving animals showed expected gains in bodyweight over the study period.
Gross pathology:
Necropsy results of animals that died during the study (treated with 2174 mg/kg) were haemorrhagic lungs, dark liver, dark kidneys, haemorragic gastric mucosa, sloughing and/or haemorrhage of the non-glandular epithelium of the stomach and haemorrhagic small and large intestines. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

Mortality data

Dose level [mg/kg]

Animal number and sex

Number of animals treated

Deaths during day of dosing (hours)

Effects noted after dosing (days)

1/2

1

2

4

1

2

3

4

5

6

7

8-14

Deaths

2174

female

3

0

0

0

0

1

1

0

0

0

0

0

0

2/3

218

female

3

0

0

0

0

0

0

0

0

0

0

0

0

0/3

male

3

0

0

0

0

0

0

0

0

0

0

0

0

0/3

Individual Clinical Observations

Dose level mg/kg

Animal Nº

and Sex

Effects Noted After Dosing (Hours)

Effects Noted During Period After Dosing (Days)

 

2174

 

 

1/2

1

2

4

1

2

3

4

5

6

7

18

9

10

11

12

13

14

1-0

Female

H

H

HD

HD

HPD

Ss

HP

H

H

 

 

 

 

 

 

 

 

 

 

1-1

Female

HS

HS

 

HSD

HSD

X

 

 

 

 

 

 

 

 

 

 

 

 

 

1-2

Female

HS

HS  

HSD

HSD

H

P

Em

Ss

Rd

RL

Wt

E

X

 

 

 

 

 

 

 

 

 

 

 

 

D: diarrhea                                                               E: pallor of extremities

Em: emaciation                                                         H: hunched posture

L: lethargy                                                               P: pilo-erection

Rd: decreased respiratory rate                                RL: labored respiration

S: increased salivation                                              Ss: red/brown stainig around snout

Wt: tiptoe gait                                                          X: animal death

 

No clinical observations were noted at the dose of 218 mg/kg.

 

Body weights

Only the animals that died during the study showed a loss of weight

 

Dose Level mg/kg

Animal Nº and sex

Body weight (g) at day

Body weight (g) at

2174

 

0

7

14

Death

1-0 Female

204

243

246

 

1-0 Female

201

 

 

175

1-0 Female

207

 

 

170

 

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The acute oral LD50 of the test substance in Sprague-Dawley rats was in the range of 500 – 1000 mg/kg of pure test substance.
Executive summary:

The Acute Oral toxicity – Acute Toxic Class method study for the test substance was performed in Sprague-Dawley CD strain rats. A stepwise procedure was used; a group of 3 females was treated with a dose of 2174 mg/kg (equivalent to 2000 mg of pure test material/kg) and 2 groups (3 females and 3 males) were treated with a dose of 218 mg/kg (equivalent to 200 mg of pure test material /kg). The test substance was administered as a solution in distilled water. An oral single dose of 10 ml/kg was given by gavage.

Clinical observations were made at 0.5, 1, 2, and 4 hours after dosing and subsequently once daily for up to 14 days. Individual body weights were recorded prior to dosing and 7 and 14 days after treatment or at death. All animals including those that died during the study were subjected to gross pathological observations. Two animals treated with 2174 mg/kg died. No mortalities were noted in animals treated with 218 mg/kg. The acute oral LD50 of the test substance in Sprague-Dawley rats was in the range of 500 – 1000 mg/kg of pure test substance.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
discriminating dose
Value:
500 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

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 October 20, 1999 to November 3, 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
Feb 1987
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source:
- Batch No.of test material: 876 TK
- Date received: 10 May 1999
- Purity: 92%
- Physical appearance: white paste

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test material was used as supplied. It was weighed out according to the animals individual body weight.
Species:
rat
Strain:
Sprague-Dawley
Remarks:
Crl: CD (SD) IGS BR
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK)
- Age at study initiation: approximately eight to twelve weeks old
- Weight at study initiation: males 206 to 232 g, and the females 214 to 225 g.
- Housing: in suspended polypropylene cages furnished with wood flakes. The animals were housed individually during the 24-hour exposure period and in groups of five, by sex, for the remainder of the study
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25 ºC
- Humidity (%): 30-70 %
- Air changes (per hr): approximately fifteen changes per hour
- Photoperiod (hrs dark / hrs light): twelve hours continuous light and twelve hours darkness
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): with cotton wool moistened with distilled water to remove any residual test material.
- Time after start of exposure: After the 24-hour contact period



TEST SITE
- Area of exposure: Back and flank of animals
- % coverage: approximating to 10% of the total body surface area.
- Type of wrap if used: Surgical gauze and self-adhesive bandage

REMOVAL OF TEST SUBSTANCE
- Washing: Residual test substance was wiped off with cotton wool moistened with distilled water
- Time after start of exposure: After a 24-hour contact period

TEST MATERIAL
- Amount applied: not specified
- Constant volume or concentration used: yes
- For solids, paste formed: yes

VEHICLE
- No vehicle used. The undiluted test substance was used.
Duration of exposure:
24-hours
Doses:
2174 mg/kg (equivalent to 2000 mg/kg bodyweight of pure test material).
No. of animals per sex per dose:
Ten animals (five males and five females)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animals were observed for deaths or overt signs of toxicity ½, 1, 2 and 4 hours after dosing and subsequently once daily for fourteen 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
- Scoring system: Scale from Draize J H (1977)
Statistics:
Mortality data was used to estimate the acute dermal median lethal dose (LD50) of the test substance
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 174 mg/kg bw
Based on:
test mat.
Remarks on result:
other: The dose 2174 mg/kg is equivalent to 2000 mg/kg bodyweight of pure test material
Mortality:
There were no deaths.
Clinical signs:
No signs of systemic toxicity were noted during the study. Very slight to well-defined erythema was noted at the treatment sites of all animals one and two days after dosing, in nine animals three days after dosing and in four animals four days after dosing. Very slight oedema was noted at the treatment sites of eight animals one day after dosing, in six animals two days and three days after dosing and in two animals four days after dosing. On the treated site of some animals a slight brown discoloration of the epidermis during days one to seven (in four animals), bleeding four days after dosing (in one animal), desquamation was noted three days (in four animals) and four days (in five animals), and crust formation after five days (in five female animals), six and seven days (in four animals each), and after eight and nine days (in three animals) was noted.
Body weight:
All animals showed expected gain in bodyweight during the study except for one female animal which showed bodyweight loss during the first week and expected gain in bodyweight during the second week.
Gross pathology:
No abnormalities were noted at necropsy.
Other findings:
Residual test material was noted at the treatment sites of all animals during the study and prevented accurate evaluation of oedema in two males one to five days after dosing.
One female showed body weight loss during the first week andd expected gain in body weight during the second week. All other animlas showed normal body weights.

Table 1: Individual clinical observations and mortality data

Dose level [mg/kg]

Animal number and sex

Effects noted after dosing (hours)

Effects noted after dosing (days)

1/2

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2174

1-0

male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-1

male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-2

male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-3

male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-4

male

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

2-4

female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0 = no signs of systemic toxicity

Table 2.1: Individual dermal reactions - males

Dose level [mg/kg]

Animal number and sex

Observation

Effects noted after dosing (days)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2174

1-0

male

Erythema

Oedema

Other

2

1

Rt

2

1

Rt

1

1

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-1

male

Erythema

Oedema

Other

2

od

RtBr

2

od

RtBr

1

od

RtBr

0

od

RtBr

0

od

RtBr

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

G

0

0

G

0

0

G

1-2

male

Erythema

Oedema

Other

2

1

Rt

1

1

Rt

1

1

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-3

male

Erythema

Oedema

Other

2

1

Rt

2

1

Rt

2

1

Rt

1

1

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

G

0

0

G

1-4

male

Erythema

Oedema

Other

2

od

Rt

2

od

Rt

1

od

Rt

0

od

RtBs

0

od

RtBr

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

G

0

0

G

0

0

G

0 = no signs of dermal irritation; G = glossy skin; Bs = Bleeding; Rt = residual test material; Br = light brown discoloration of the epidermis; od = adverse reactions prevent accurate evaluation of oedema

Table 2.2: Individual dermal reactions - females

Dose level [mg/kg]

Animal number and sex

Observation

Effects noted after dosing (days)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2174

2-0

female

Erythema

Oedema

Other

2

1

Rt

1

1

Rt

1

1

RtD

1

1

RtD

0

0

RtCf

0

0

Cf

0

0

Cf

0

0

Cf

0

0

Cf

0

0

G

0

0

G

0

0

G

0

0

G

0

0

G

2-1

female

Erythema

Oedema

Other

2

1

RtBr

2

1

RtBr

2

1

RtBrD

1

0

RtBrD

0

0

RtBrCf

0

0

Cf

0

0

Cf

0

0

Cf

0

0

Cf

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-2

female

Erythema

Oedema

Other

2

1

Rt

2

0

Rt

1

0

RtD

0

0

RtD

0

0

RtCf

0

0

Cf

0

0

Cf

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-3

female

Erythema

Oedema

Other

2

1

RtBr

2

1

RtBr

2

1

RtBrD

2

0

RtBrD

0

0

RtBrCf

0

0

BrCf

0

0

BrCf

0

0

Cf

0

0

Cf

0

0

G

0

0

G

0

0

G

0

0

G

0

0

G

2-4

female

Erythema

Oedema

Other

2

1

Rt

1

0

Rt

0

0

Rt

0

0

D

0

0

Cf

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0 = no signs of dermal irritation; D = desquamation; Br = light brown discoloration of the epidermis; G = glossy skin; Cf = crust formation; Rt = residual test material

Table 3: Individual bodyweight and weekly changes

 

Doselevel

mg/kg

Animal Number and Sex

Bodyweight

(g) atDay

Bodyweight Gain (g)

During Week

 

 

0

7

14

1

2

2174

1-0 Male

206

238

286

32

48

1-1 Male

228

285

336

57

51

1-2 Male

232

280

345

48

65

1-3Male

220

257

312

37

55

1-4 Male

215

201

322

46

61

2-0 Female

214

215

234

1

19

2-1 Female

220

225

236

5

11

2-2 Female

225

220

245

-5

25

2-3 Female

224

235

265

11

30

2-4 Female

225

226

250

1

24

 

Interpretation of results:
GHS criteria not met
Conclusions:
The acute dermal median lethal dose (LD50) of the test substance in the Sprague-Dawley CD strain rat was found to be greater than 2174 mg/kg bodyweight (equivalent to 2000 mg/kg bodyweight of pure test material).
Executive summary:

The Acute Dermal Toxicity assay for the test substance was performed in Sprague-Dawley CD rats. One female group and one male group, of 5 animals each one, were treated with a single dose of 2174 mg/kg bodyweight, equivalent to 2000 mg/kg bodyweight of pure test material (Limit test). The test material was applied uniformly to an area of shorn skin (approximating to 10% of the total body surface area). A piece of surgical gauze was placed over the treatment area and semi-occluded with a piece of self-adhesive bandage. After the 24-hour contact period the bandage was carefully removed and the treated skin and surrounding hair wiped with cotton wool moistened with distilled water. Clinical observation was conducted at 1/2,1, 2 and 4 hours after dosing and subsequently once daily for fourteen days. Body weight was controlled before the administration, on day 0 and on days 7 and 14 after the administration. All animals were subjected to gross necropsy. No test substance related effects were noted from clinical observations or post-mortem examination. The acute dermal median lethal dose (LD50) of the test substance in the Sprague-Dawley CD strain rat was found to be greater than 2174 mg/kg bodyweight (equivalent to 2000 mg/kg bodyweight of pure test material).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Additional information

The Acute Oral toxicity – Acute Toxic Class method study for the test substance was performed in Sprague-Dawley CD strain rats. A stepwise procedure was used; a group of 3 females was treated with a dose of 2174 mg/kg (equivalent to 2000 mg of pure test material/kg) and 2 groups (3 females and 3 males) were treated with a dose of 218 mg/kg (equivalent to 200 mg of pure test material /kg). The test substance was administered as a solution in distilled water. An oral single dose of 10 ml/kg was given by gavage.

Clinical observations were made at 0.5, 1, 2, and 4 hours after dosing and subsequently once daily for up to 14 days. Individual body weights were recorded prior to dosing and 7 and 14 days after treatment or at death. All animals including those that died during the study were subjected to gross pathological observations. Two animals treated with 2174 mg/kg died. No mortalities were noted in animals treated with 218 mg/kg.The acute oral LD50 of the test substance in Sprague-Dawley rats was calculated in the range of 500 – 1000 mg/kg of pure test substance.

The Acute Dermal Toxicity assay for the test substance was performed in Sprague-Dawley CD rats. One female group and one male group, of 5 animals each one, were treated with a single dose of2174 mg/kg bodyweight, equivalent to 2000 mg/kg bodyweight of pure test material(Limit test).The test material was applieduniformly to an area of shorn skin (approximating to 10% of the total body surface area). A piece of surgical gauze was placed over the treatment area and semi-occluded with a piece of self-adhesive bandage. After the 24-hour contact period the bandage was carefully removed and the treated skin and surrounding hair wiped with cotton wool moistened with distilled water.Clinical observation was conducted at 1/2,1, 2 and 4 hours after dosing and subsequently once daily for fourteen days.Body weight was controlled before the administration, on day 0 and on days 7 and 14 after the administration. All animals were subjected to gross necropsy.No test substance related effects were noted from clinical observations or post-mortem examination.The acute dermal median lethal dose (LD50) of the test substance in the Sprague-Dawley CD strain rat was found to be greater than 2174 mg/kg bodyweight (equivalent to 2000 mg/kg bodyweight of pure test material).

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for the purpose of classification. Based on the criteria laid down in Regulation (EC) No.1272/2008, as amended for the second time in Directive EC 286/2011, the test substance is classified for acute toxicity with category 4 (H302).