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Description of key information

Oral (OECD 422, read across): NOAEL female rat = 300 mg/kg bw/day

Oral (OECD 422, read across): NOAEL male rat = 1000 mg/kg bw/day

Oral (OECD 407, read across): NOAEL, male and female rat >1000 mg/kg bw/day

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
Refer to analogue justification provided in IUCLID section 13.
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Key result
Dose descriptor:
NOAEL
Effect level:
300 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
body weight and weight gain
food consumption and compound intake
Remarks on result:
other:
Remarks:
Source: CAS 59231-34-4
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male
Remarks on result:
not determinable due to absence of adverse toxic effects
Remarks:
Source: CAS 59231-34-4
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Remarks:
Source: CAS 91031-48-0
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Remarks:
Source: CAS 3687-46-5
Key result
Critical effects observed:
no
Conclusions:
CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008.
Endpoint:
short-term repeated dose toxicity: oral
Remarks:
combined repeated dose and reproduction / developmental screening
Type of information:
experimental study
Adequacy of study:
key study
Study period:
01 Nov - 13 Dec 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
22 Mar 1996
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Behoerde fuer Soziales, Familie, Gesundheit und Verbraucherschutz; Hamburg, Germany
Limit test:
no
Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories Research Models and Services Germany GmbH, Sulzfeld, Germany
- Age at study initiation: males: 50 days; females: 60 days
- Mean weight at study initiation: males: 248.8 to 298.7 g; females: 195.2 to 228.1 g
- Fasting period before study: no
- Housing: single housing in MAKROLON cages (type III plus)
- Diet: commercial ssniff R-Z V1324 (ssniff Spezialdiäten GmbH, Soest, Germany); ad libitum
- Water: tap water; ad libitum
(Analyses of diet and water was performed.)
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 50, 150 and 500 mg/mL
- Amount of vehicle (if gavage): 2 mL/kg
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
For the analysis of the test item-vehicle mixtures samples were taken at the following time points and stored at ≤ -20°C until analysis:
Start of treatment period; immediately after preparation of the test item-vehicle mixtures; 8 and 24 hours after storage of the test item preparations at room temperature; end of treatment period; during treatment with the test item always before administration to the last animal of the dose level group
The following parameters were determined: linearity, accuracy, precision, sensitivity, specificity, stability at +2°C to +8°C or -20°C (0, 24, 72 and 168 hours)
Duration of treatment / exposure:
Males: The daily administration of the test item was started two weeks before mating and lasted until test day 35, which was one day before sacrifice.
Females: The daily administration of the test item was started two weeks before mating and continued to at least day 3 of lactation.
Maximum: 56 days of treatment
Frequency of treatment:
once daily; 7 days/week
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: based on a 14-day range-finding study (Leuschner, 2012)
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: before and after dosing
- Cage side observations checked: skin/fur, eyes, mucous membranes, respiratory and circulatory systems, somatomotor activity and behaviour patterns

MORTALITY AND CLINICAL SIGNS
- Time schedule: at least once daily (the frequency was increased when signs of toxicity were observed); deaths were recorded twice daily (animals which died or were sacrificed during the study were necropsied as soon as possible after exitus)

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once before the first exposure (to allow within-subject comparisons) and once a week thereafter
- Paramters: changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions and autonomic activity (e.g. lacrimation, pilo-erection, pupil size, and unusual respiratory pattern); changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies (e.g. excessive grooming, repetitive circling), difficult or prolonged parturition or bizarre behaviour (e.g. self-mutilation, walking backwards)

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: daily by visual appraisal

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations:
- Dose groups that were examined: all dose groups

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at the end of the pre-mating period
- Anaesthetic used for blood collection: Yes (ether)
- Animals fasted: Yes, overnight
- How many animals: 5/sex/dose
- Parameters: haemoglobin, erythrocytes, leucocytes, differential blood count (absolute/relative), reticulocytes, platelets, haematocrit, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, thromboplastin time, activated partial thromboplastin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the end of the pre-mating period
- Animals fasted: Yes, overnight
- How many animals: 5/sex/dose
- Parameters examined: albumin, globulin, albumin/globulin ratio, bile acids, bilirubin, cholesterol (total), creatinine, glucose, urea, total protein, calcium, chlorid, potassium, sodium, alanine aminotransferase (ALAT), alkaline phosphatase (AP), aspartate aminotransferase (ASAT)

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: two hours after dosing shortly before scheduled sacrifice in 5 males per group (day 35); two hours after dosing during lactation, shortly before scheduled sacrifice in 5 females per group (day 39-56)
Screening of assessment were conducted as described on the following pages in five males and five females randomly selected from each group.
- Dose groups that were examined: all dose groups
- Battery of functions tested: sensory reactivity to stimuli of different types (e.g. auditory, visual and proprioceptive stimuli) (based on Gad), as well as the assessment of grip strength (Meyer) and motor activity

OTHER: Qualitative sperm analysis was performed.
(for further reproduction parameters see respective study entry (chapter 7.8.1))
Males were sacrificed on day 36, females were sacrifices on day 4 post-partum or shorty thereafter.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
- Organ weights: epididymes and testicles (all males); adrenal gland, brain, heart, kidney, liver, spleen, thymus (5/sex/dose)
- Fixation: epididymis, gross lesions, mammary gland, ovary, prostate, seminal vesicle, testicle, uterus (incl. cervix and oviducts), vagina (all animals); adrenal gland, bone marrow (os femoris), brain (cerebrum, cerebellum, brain stem), heart (left and right ventricle, septum), intestine, small (duodenum, jejunum, ileum, incl. Peyer's patches, Swiss roll method), intestine, large (colon, rectum), kidney and ureter, liver, lungs (with mainstem bronchi and bronchioles), preserved by inflation with fixative and then immersion, lymph node (1 cervical, 1 mesenteric), nerve (sciatic), oesophagus, spinal cord (3 sections), spleen, stomach, thyroid (incl. parathyroids), thymus, tissue masses or tumours (incl. regional lymph nodes), tongue (incl. base), trachea (incl. larynx), urinary bladder (5/sex/dose)
HISTOPATHOLOGY: Yes, all organs that were included for fixation (5/sex of control and high dose group)
Statistics:
STUDENT's t-test (p ≤ 0.01): all numerical functional tests
Multiple t-test based on DUNNETT (p≤0.05 and p ≤ 0.01): body weight, food consumption, haematology, clinical chemistry, absolute and relative organ weights
For all numerical values homogeneity of variances was tested by using the BARTLETT chi-square test. If the variances were homogeneous, the DUNNETT test (p ≤ 0.01) was used to compare the experimental groups with the control group. In case of heterogeneity of variances, the STUDENT's t-test was carried out; limit of significance was p ≤ 0.01.
Clinical signs:
no effects observed
Description (incidence and severity):
Piloerection was seen in 1 female of the high dose group on day 2-4 of lactation.
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
statistically significant decrease (-9.4%) in body weight in females during lactation at 1000 mg/kg bw/day
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
In the high dose group (females: gestation/lactation period) a statistically significant reduction in food consumption by 21.7% was noted.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
An decrease of ASAT activity was seen in high dose females on day 15. This effects was considered to be due to the relative low or high value observed for the control group and not to be test item related.
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
All effects observed (slight increase in absolute and relative liver weight in males) were still within the historical control data of the laboratory and thus not of toxicological relevance.
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Other effects:
no effects observed
Description (incidence and severity):
The qualitative sperm staging revealed no test item-related specific spermatogenic changes in the male animals from the high dose group (1000 mg/kg b.w./day).
Key result
Dose descriptor:
NOAEL
Effect level:
300 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
body weight and weight gain
food consumption and compound intake
Key result
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Critical effects observed:
no

Individual body weights of females during the pre-mating and lactation period.

Control group

Day(s) relative to start

 

1

8

15

11

196

209

212

12

217

228

243

13

210

220

213

14

217

234

244

15

3211

215

232

16

195

208

197

17

205

224

239

18

212

238

233

19

224

236

259

20

200

218

214

Mean

209

223

229

SD

9

10

19

1000 mg/kg bw

Day(s) relative to start

 

1

8

15

71

200

210

225

72

210

231

234

73

206

234

247

74

210

222

235

75

194

219

218

76

218

243

223

77

214

230

227

78

222

225

210

79

198

200

201

80

203

223

231

Mean

207

224

225

SD

9

12

13

Control group

Day(s) relative to littering

 

 

1

4

11

286

309

12

323

330

13

325

311

14

331

327

15

311

322

16

282

302

17

309

327

18

312

328

19

315

331

20

300

329

Mean

309

322

SD

16

10

1000 mg/kg bw

Day(s) relative to littering

 

 

1

4

71

270

281

72

339

301

73

289

309

75

289

317

77

253

247

78

280

271

79

290

296

80

293

308

Mean

288

291

SD

24

23

Relative food consumption of females between day 1 and 4 of lactation

Control group

1000 mg/kg bw

 

 

11

122

71

115

12

91

72

96

13

105

73

63

14

107

75

110

15

106

77

30

16

121

78

63

17

114

79

98

18

100

80

110

19

101

 

 

20

126

 

 

Mean

109

Mean

86

SD

11

SD

30

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
16 Nov 1982 - 19 Jan 1983
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
No monitoring of clinical signs outside the home cage, no neurobehavioural examinations.
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Deviations:
yes
Remarks:
no monitoring of clinical signs outside the home cage, no neurobehavioural examinations
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
other: Wistar Han.
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Züchter Zentralinstitut für Versuchstiere, Hannover, Germany
- Age at study initiation: approximately 4 weeks
- Weight at study initiation: 50-78 g (males); 50-82 g (females)
- Housing: 2-3 animals/sex/cage in Macrolon cages (type III) with wood shavings (Arwi-Center, Essen, Germany)
- Diet: Altromin 1324 DK pelleted, nitrosamine-poor diet (Fa. Altromin GmbH, Lage, Germany), ad libitum
- Water: tap water, ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (ºC): 21-22
- Humidity (%): 49-65
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: The dosing solutions were prepared daily prior to administration. The application volumes were adapted weekly to the current body weights.

VEHICLE
- Concentration in vehicle: 2, 10 and 20% in olive oil for the 100, 500 and 1000 mg/kg bw/day groups, respectively
- Amount of vehicle (if gavage): 5 mL/kg bw
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
28 days (21 doses) and 28 days post-exposure observation period (satellite control and treated groups)
Frequency of treatment:
Daily, 5 days/week
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
500 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10 (main groups, control and all dose levels)
5 (satellite groups, control and all dose levels)
Control animals:
yes, concurrent vehicle
Details on study design:
- Post-exposure recovery period in satellite groups: 28 days. A satellite groups was included in the control group and each dose group.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations examined: mortality, clinical signs

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION : Yes. Determined per cage per week.

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION: Yes. Determined per cage per week.

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: prior to necropsy
- Dose groups that were examined: all dose groups

HAEMATOLOGY: Yes
- Time schedule for collection of blood: prior to necropsy
- Anaesthetic used for blood collection: Yes, ether
- Animals fasted: No data
- How many animals: all dose groups
- Parameters examined: haematocrit, haemoglobin content, red blood cell count, white blood cell count, mean cell volume, thrombocyte cell count, differential blood count

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: prior to necropsy
- Animals fasted: No data
- How many animals: all dose groups
- Parameters examined: urea, creatinine, glucose, sodium, potassium, calcium, inorganic phosphorus, alkaline phosphatase, serum alanine aminotransferase, serum aspartate aminotransferase, gamma glutamyl transpeptidase, chloride, albumin, total protein, cholesterol

URINALYSIS: Yes
- Time schedule for collection of urine: at the end of the study period
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters examined: pH, proteins, glucose, blood cells, bacteria, leucocytes, erythrocytes, urea, sodium oxide, urobilin, ket., specific gravity, epi., trip.

NEUROBEHAVIOURAL EXAMINATION: No

OTHER: faeces were collected at the end of the study and examined for parasites, appearance and amount.
Sacrifice and pathology:
GROSS PATHOLOGY:
Yes. Gross pathology was performed in all animals, including the satellite group animals. The absolute and relative weight of the following organs was determined: thyroid, adrenals, thymus, spleen, heart, kidneys, brain, testes, ovaries and liver.

HISTOPATHOLOGY:
Yes, in all animals in the control and 1000 mg/kg bw/day groups the following organs were examined: brain (cerebrum, cerebellum), eye, tongue, salivary glands, auxillary lymph nodes, oesophagus, trachea, heart, aorta, lungs, bronchial lymph nodes, liver, spleen, kidneys,
urethra, pancreas, thymus, thyroid, parathyroid, adrenals, peripheral nerve, forestomach, stomach, duodenum, jejunum, ileum, colon, caecum, rectum, anal mucosa, Peyer's patches, mesenterial lymph node, gall bladder, urinary bladder, vagina, uterus, ovary, prostate, testes, epididymes, prostate + seminal vesicles with coagulating glands, skin, muscle, skin lymph nodes, mammary gland.

The target organ of the main groups was the liver, which was examined in greater detail in 3 male and 3 female animals in the control and 1000 mg/kg bw/day groups. In the satellite groups, the target organs (lungs, pancreas, maxillary lymph node, salivary gland and forestomach) were examined in 3 females and males from the control and 1000 mg/kg bw/day satelllite groups.
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
The body weight was statistically significantly lower in the male 100 mg/kg bw/day group in week 0 and 1. As the rats weighed, on average, less from the start of the study, this reflects a mistake in standardising the group and is not related to the treatment (see Table 1).
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
The food consumption in females and males administered 1000 mg/kg bw/day was statistically significantly increased in week 2, while in the males in this dose group the intake was reduced in week 3. Both males and females in the 500 mg/kg bw/day group had a statistically significant increase in food consumption in week 2. These effect were transient and were considered not to be treatment-related (see Table 1).
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
effects observed, non-treatment-related
Description (incidence and severity):
The water intake was statistically significantly increased in males in the highest dose group in week 2. For males administered 100 mg/kg bw/day the intake was decreased in week 1 and 2. In the female groups, the water consumption was reduced with statistical significance in females administered 100, 500 and 1000 mg/kg bw/day in week 2, week 3 and 4, and week 3, respectively. These effect were transient and were considered not to be substance-related (see Table 1).
Ophthalmological findings:
no effects observed
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Statistically significant decreases in haemoglobin levels were observed in all dose groups. As the changes were not dose-related and no other relevant effects were seen in histopathological parameters, this is not considered to be a toxicologically relevant effect. A reduction in the haematocrit level and red blood cell count in the 500 mg/kg bw/day females was considered to be incidental as the effect was only observed at one dose level and in one sex.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
The inorganic phosphorus level was increased with statistical significance in all male dose groups and in the 100 and 500 mg/kg bw/day female group. The effect was not dose-related in the males and not observed at the highest dose level in females, therefore it was not considered to be substance-related. The decrease in the alkaline phosphatase level in females in the 500 mg/kg bw/day group was deemed to be incidental and not substance-related.
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
In females administered 1000 mg/kg bw/day, the absolute kidney weight was increased. The relative weight was not affected and the change was seen only in one sex, leading to the conclusion that this effect had no toxicological relevance. The relative heart weight in males administered 500 and 1000 mg/kg bw/day was reduced slightly, but with statistical significance, while the absolute heart weight was increased in the 500 mg/kg bw/day group. The highest male dose group was not affected and no related effects were noted in the histopathological examination. Therefore, this effect was considered of no toxicological relevance. Some statistically significant changes in organ weights of the thyroid, spleen and adrenal glands were noted in males or females in the 100 or 500 mg/kg bw/day groups. As the changes were either absolute or relative weight, only in one sex and not observed in the highest dose level, they were not considered to be treatment-related. No treatment-related changes were noted in the salivary gland, lungs, pancreas or stomach in the satellite animals.
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
No substance-related effects were observed on organs or tissues during the histopathological examination. The mucosa of the forestomach in control and treated groups showed degenerative changes or signs of inflammation. 2/10 males and 3/10 females in the control groups plus 2/10 females in the 1000 mg/kg bw/day group had (suspected) hyperplasia; and 7/10 control males, 7/10 control females, 5/10 1000 mg/kg bw/day males and 7/10 1000 mg/kg bw/day females showed focal eosinophilic infiltration of the forestomach. These effects were possibly caused by the repeated use of a stomach tube for the dosing by gavage. As humans do not have a forestomach, this effect is not relevant to human exposure.
Histopathological findings: neoplastic:
not examined
Other effects:
no effects observed
Description (incidence and severity):
Eggs from a parasite were detected in the faeces of 1 female in the high-dose group. No other significant effects were seen on the faeces appearance or amount.
Key result
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Critical effects observed:
no

Table 1: Data of selected parameters determined in the study for all groups:

Food consumption (g/animal/d), group mean values

 

 

 

 

week

group***, sex (m/f)

 

 

 

 

 

 

 

1 m

2 m

3 m

4 m

1 f

2 f

3 f

4 f

1

18

16

18

18

14

15

14

15

2

19

18

21**

22**

14

14

15**

16*

3

22

21

21

21*

16

17

15

16

4

17

17

19

19

13

13

13

14

Water consumption (mL/animal/d), group mean values

 

 

 

 

week

group, sex (m/f)

 

 

 

 

 

 

 

1 m

2 m

3 m

4 m

1 f

2 f

3 f

4 f

1

29

25*

27

29

27

24

24

24

2

28

25**

30

31*

28

22**

26

27

3

33

29

30

30

32

27

26*

25*

4

30

25

27

29

29

28

24*

24

Body weights and weight gain (g)

 

 

 

 

 

 

week

group, sex (m/f)

 

 

 

 

 

 

 

1 m

2 m

3 m

4 m

1 f

2 f

3 f

4 f

0

67

60*

64

67

64

63

60

63

1

107

96**

105

108

96

96

94

95

2

147

137

147

150

123

124

122

124

3

189

178

191

196

146

147

146

147

4

220

208

227

229

161

162

163

173

gain week 0-4

153

148

163

162

97

99

103

110

Selected haematology and clinical chemistry parameters (week 4)

 

 

 

 

 

 

group, sex (m/f)

 

 

 

 

 

 

 

1 m

2 m

3 m

4 m

1 f

2 f

3 f

4 f

haemoglobin mmol/L

10.9

10.1**

9.9**

9.8**

10.6

9.9**

9.9**

9.8**

red blood cell count T/L

7.2

6.9

6.6

6.8

6.9

6.4

6.0*

6.5

haematokrit l/L

0.39

0.38

0.36

0.37

0.36

0.34

0.32**

0.35

alkaline phosphatase U/L

371

356

390

361

247

211

196*

206

inorganic phosphate

3.1

2.9*

2.7**

2.8*

2.6

2.4*

2.4

2.5

Organ weights after 4 weeks

 

 

 

 

 

 

 

group, sex (m/f)

 

 

 

 

 

 

 

1 m

2 m

3 m

4 m

1 f

2 f

3 f

4 f

thyroid absolute (mg)

25

22*

23

23

21

23

19

22

spleen absolute (g)

0.57

0.59

0.63*

0.63

0.46

0.45

0.46

0.46

heart absolute (g)

0.80

0.73*

0.78

0.80

0.61

0.61

0.58

0.64

kidney absolute (g)

1.66

1.58

1.73

1.80

1.19

1.22

1.19

1.28*

thyroid relative (% of body weight)

0.010

0.010

0.009*

0.009

0.013

0.014

0.011

0.013

adrenal relative (% of body weight)

0.025

0.025

0.024

0.025

0.046

0.044

0.039*

0.045

heart relative (% of body weight)

0.33

0.32

0.31**

0.31*

0.36

0.36

0.34

0.37

* level of significance 95% in comparison with control value

** level of significance 99% in comparison with control value

*** 1 = control, 2 = 100 mg/kg bw/day, 3 = 500 mg/kg bw/day, 4 = 1000 mg/kg bw/day

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
(analytical purity of test substance not specified; no urine and neurobihavioural examinations)
Qualifier:
according to guideline
Guideline:
other: 87/302/EWG, Annex, Part B
Deviations:
yes
Remarks:
analytical purity of test substance not specified; no urine and neurobihavioural examinations
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Deviations:
yes
Remarks:
analytical purity of test substance not specified; no urine and neurobihavioural examinations
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga, Sulzfeld, Germany.
- Age at study initiation: ca. 40 days
- Weight at study initiation: 143 - 168 g (153.4 g mean body weight)
- Housing: 2-3 animals per Makrolon Type III cage on softwood bedding (ARWI-Center, Essen, Germany).
- Diet: pelleted Altromin 1324, Altromin, Lage, Germany, ad libitum
- Water: tap water, ad libitum
- Acclimation period: 11 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23
- Humidity (%): 48-60
- Air changes (per hr): nodata, 80-490 Lux (the cages were rotated weekly in the rack)
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: March 28th, 1989 - April 28th, 1989
Route of administration:
oral: gavage
Vehicle:
peanut oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
The application volume was 5 mL/kg bw.
The dosing solutions were prepared daily immediately before application.
The following concentrations were prepared: 100 mg/ kg bw /day: 2%; 500 mg/kg bw/day: 10%, 1000 mg/kg bw/day: 20%
The total number of applications was 23 or 24 (depending on the day of necropsy).
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
28 days
Frequency of treatment:
once daily, 5 days/week
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
500 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Rationale for selecting satellite groups: Additionally 5 male and 5 female animals per dose were dosed with 0 and 1000 mg/kg/day to determine the reversibility of possible compound-related findings (recovery group).
- Post-exposure recovery period in satellite groups: 27 days
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice a day, 5 days a week for mortality and clinical signs

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION: Yes
- Time schedule for examinations: weekly

WATER CONSUMPTION: Yes
- Time schedule for examinations: weekly

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: at study termination
- Dose groups that were examined: control group and highest dose group

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at study termination
- Anaesthetic used for blood collection: not reported
- Animals fasted: No data
- How many animals: all animals
- Parameters checked: RBC, HCT, MCV, HGB, WBC, PLT, differential blood count (banded neutrophils, segmented neutrophils, lymphocytes, eosinophils, monocytes, basophils, myelocytes).

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at study termination
- Animals fasted: No data
- How many animals: all animals
- Parameters checked: Urea, Creatinine, Sodium, Potassium, Glucose, Calcium, ASAT, ALAT, AP, gamma-GT, Bilirubin, Chloride, total protein, total Cholesterol.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

ORGAN WEIGHTS:
Absolute and relative organ weights were determined for the following:
Brain, testes, heart, liver, spleen, adrenals, kidneys, thymus.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes, all animals
HISTOPATHOLOGY: Yes, animals of the control and highest dose group.
Following organs were examined microscopically:
Aorta thoracica, eyes, large and small intestinum, glandular stomach, cerebrum, urinary bladder, skin, heart, testes, pituitary, cerebellum, liver, trachea, lung, maxillary lymph nodes, mesenterial lymph nodes, spleen, epididymides, adrenal, peripheral nerv, kidney, ovaries, pancreas, prostate, vesicular gland, thymus, salivary gland, oesophagus, sceletal muscle, thymus, uterus, stomach, tongue.
Statistics:
T-tests according to Sachs and Dunnett; steel-test
Clinical signs:
no effects observed
Description (incidence and severity):
No symptoms were noted as compound-related effects.
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
effects observed, non-treatment-related
Description (incidence and severity):
The mean water intake of the male groups 2 and 4 showed a slight increase which was not considered to be compound-related.
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Description (incidence and severity):
Some observations like hydrometra, hydronephrosis, discolouration of the thymus and one suspicion of hydrocephalus internus were considered to be spontaneous.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Details on results:
The histological examination of the organs of the recovery group was not performed, because in the main groups no target organ was evaluated.
Key result
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other:
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Critical effects observed:
no
Conclusions:
According to the described study, a daily administration of 2-Ethylhexylstearate up to 1000 mg/kg bodyweight/day for 28 days caused no cumulative-systemic toxicity to rats. The NOAEL was found to be 1000 mg/kg bw/day.
Executive summary:

2-Ethylhexylstearate was tested for systemic toxicity at repeated doses of 0 (group 1), 100 (group 2), 500 (group 3) and 1000 (group 4) mg/kg body weight/day. The compound was administered daily by gavage over a period of 28 days. 10 male and 10 female rats were used for each dose. In addition to the groups 1 and 4, 5 male and 5 female animals were used to determine the reversibility of possible compound-related findings (recovery group).

 

All doses applied were tolerated without lethality. No symptoms were noted as compound-related effects. The mean food consumption of all groups was comparable to the control. The mean water intake of the male groups 2 and 4 showed a slight increase which was not considered to be compound-related. The total body weight gain was comparable to control in all male and female test groups. The haematological examinations showed no compound-related effects. The clinical chemistry showed no compound-related effects. The examination of the eyes by slit lamp microscope showed no compound-related effects. The absolute and relative organ weights showed no compound-related effects. The macroscopical examination of the organs displayed no compound-related effects. Some observations like hydrometra, hydronephrosis, discolouration of the thymus and one suspicion of hydrocephalus internus (animal No. 67) were considered to be spontaneous. The histological examination of the organs of all groups displayed no compound-related effects.The histological examination of the organs of the recovery group was not performed, because in the main groups no target organ was evaluated.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
300 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The available information comprises adequate and reliable (Klimisch score 1-2) studies from reference substances with similar structure and intrinsic properties. Read-across is justified based on common precursors and breakdown products of hydrolysis and consistent trends in environmental fate, ecotoxicological and toxicological profile (refer to the endpoint discussion for further details). The selected studies are thus sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.6, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Read Across Justification

There are no data on the repeated dose toxicity of Fatty acids, C16-18, isononyl esters (CAS 91031-57-1). The assessment was therefore based on studies conducted with analogue substances as part of a read across approach, which is in accordance with Regulation (EC) No. 1907/2006, Annex XI, 1.5. For each specific endpoint the source substance(s) structurally closest to the target substance is/are chosen for read-across, with due regard to the requirements of adequacy and reliability of the available data. Structural similarities and similarities in properties and/or activities of the source and target substance are the basis of read-across. A detailed justification for the analogue read-across approach is provided in the technical dossier (see IUCLID Section 13).

Repeated dose toxicity, oral, subacute

CAS 59231-34-4

A combined repeated dose toxicity and reproduction/developmental toxicity screening study was performed according to OECD guideline 422 and under GLP conditions (key study, 2013) with Isodecyl oleate (CAS 59231-34-4). 10 rats/sex/dose were administered 0, 100, 300 and 1000 mg/kg bw/day once daily for 35 days (males) and up to 56 days (females) via oral gavage. The application started two weeks before mating and ended on the day of or one day before sacrifice. Day of sacrifice was on day 35 for the male rats and on lactation day 3 or shortly thereafter for the female rats. There was no mortality during the study period. No toxicologically relevant clinical signs were observed. A statistically significant decrease in body weight (‑9.4%) and food consumption (‑21.7%) was noted in females at 1000 mg/kg bw/day during lactation. There were no toxicologically relevant effects on water consumption and organ weights. No treatment-related effects were noted on the ophthalmology -, haematology - and clinical biochemistry parameters. The macroscopic examination at autopsy and subsequent histopathological examination did not reveal any treatment-related changes. The NOAEL for systemic toxicity was considered to be 300 mg/kg bw/day for female rats and 1000 mg/kg bw/day for male rats.

CAS 91031-48-0

A 28-day oral repeated dose toxicity study was performed with Fatty acids, C16-18, 2-ethylhexyl esters (CAS 91031-48-0) similar to OECD 407 and under GLP conditions (supporting study, 1992). Ten Sprague-Dawley rats per sex and dose were administered once daily (5 days a week) 0, 100, 500 and 1000 mg/kg bw/day by gavage, for 28 consecutive days. There was no mortality and no toxicologically relevant clinical signs were observed during the study period. No significant differences in body weight, body weight gain, food consumption, water consumption and organ weight were noted between the control group and treatment groups. No treatment-related effects on the ophthalmology-, hematology- and clinical biochemistry parameters were observed. No treatment-related changes were noted during the gross pathology and histopathology examinations. Based on the absence of adverse toxic effects the NOEL for systemic toxicity was 1000 mg/kg bw/day.

CAS 3687-46-5

A 28-day oral repeated dose toxicity study was performed with Decyl oleate (CAS 3687-46-5) similar to OECD 407 (supporting study, 1987). Ten Wistar rats per sex and dose and 5 per sex and dose satellite rats were administered once daily (5 days a week) 0, 100, 500 and 1000 mg/kg bw/day by gavage, for 28 consecutive days. There was no mortality and no toxicologically relevant clinical signs were observed during the study period. No significant differences in body weight, body weight gain, food consumption, water consumption and organ weight were noted between the control group and treatment groups. No treatment-related effects on the ophthalmology-, hematology- and clinical biochemistry parameters were observed. No treatment-related changes were noted during the gross pathology and histopathology examinations. Based on the absence of adverse toxic effects the NOAEL for systemic toxicity was 1000 mg/kg bw/day.

Overall conclusion for repeated dose toxicity

The data for the source substance showed no toxicologically relevant effects up to and including the recommended limit values. Therefore, as the available data did not identify any hazard for repeated dose toxicity, Fatty acids, C16-18, isononyl esters (CAS 91031-57-1) is not considered to be hazardous following repeated exposure.

Justification for classification or non-classification

According to Article 13 of Regulation (EC) No. 1907/2006 "General Requirements for Generation of Information on Intrinsic Properties of substances", information on intrinsic properties of substances may be generated by means other than tests e.g. from information from structurally related substances (grouping or read-across), provided that conditions set out in Annex XI are met. Annex XI, "General rules for adaptation of this standard testing regime set out in Annexes VII to X” states that “substances whose physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity may be considered as a group, or ‘category’ of substances. This avoids the need to test every substance for every endpoint". Since the analogue concept is applied to Fatty acids, C16-18, isononyl esters (CAS 91031-57-1), data will be generated from data for reference source substance(s) to avoid unnecessary animal testing. Additionally, once the analogue read-across concept is applied, substances will be classified and labelled on this basis.

Therefore, based on the analogue read-across approach, the available data on repeated dose toxicity do not meet the classification criteria according to Regulation (EC) 1272/2008 and are therefore conclusive but not sufficient for classification.