Registration Dossier

Toxicological information

Developmental toxicity / teratogenicity

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Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2017

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
2001
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid

Test animals

Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS srl, San Pietro al Natisone (UD), Italy
- Age at arrival: 9 weeks (females), 11 weeks (males)
- Weight at arrival: 320-339 g (males), 190-214 g (females)
- Fasting period before study: no
- Housing: during pre-pairing phase and after mating: max. 5 of one sex to a cage, in polisulphone solid bottomed cages; during mating: 1 m + 1 f in clear polisulphone cages with a stainless steel mesh lid and floor
- Diet (e.g. ad libitum): laboratory rodent diet (4 RF 21, Mucedola S.r.l.) ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: approximately 3 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±2
- Humidity (%): 55±15
- Air changes (per hr): 15-20
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: 0.5% carboxymethyl cellulose
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The formulations were prepared daily.

VEHICLE
- Concentration in vehicle: 10, 30 and 100 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Formulations of the test item were prepared as suspensions in 0.5% CMC. Concentration and homogeneity of the low and high dose level were assessed by taking six analytical aliquots in different positions. For the intermediate levels, only concentration was assessed by taking two different analytical aliquots.
28 hour stability at room temperature and a 8 day stability at +4°C were verified in the range from 10 to 100 mg/mL.
The results of the analyses were within the acceptability limits (85-115% for concentration and CV < 10% for homogeneity).
Details on mating procedure:
- Impregnation procedure: cohoused
- M/F ratio per cage: 1:1
- Length of cohabitation: ???
- Further matings after two unsuccessful attempts: no
- Verification of same strain and source of both sexes: yes
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy
Duration of treatment / exposure:
from day 6 through day 19 post coitum
Frequency of treatment:
daily
Doses / concentrationsopen allclose all
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:
24
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: based on information from previous study (reproduction/developmental toxicity screening test)
- Rationale for animal assignment (if not random): computerised stratified randomisation to give approximately equal initial group mean body weights

Examinations

Maternal examinations:
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: days 0, 6, 9, 12, 15, and 20 post coitum

FOOD CONSUMPTION: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
6, 9, 12, 15 and 20 post coitum

WATER CONSUMPTION: No

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day #20
- Organs examined: abnormalities

Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: gross evaluation of placentae
Fetal examinations:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: Yes: half per litter
Statistics:
For continuous variables the significance of the differences amongst group means was assessed by Dunnett’s test or a modified t test, depending on the homogeneity of data. Statistical analysis of non-continuous variables was carried out by means of the Kruskal-Wallis test and intergroup differences between the control and treated groups assessed by a non-parametric version of the Williams test.

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
no effects observed
Description (incidence and severity):
No signs of toxicological significance were noted during the study and no signs of reactions to treatment were observed during the dosing period. The scab on the neck seen in one high dose female during the last day of study was considered of no relevance.
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No relevant differences were noted in body weight and body weight gain of females during the study, between control and treated groups. No importance was attributed to the slight statistically significant increase in body weight gain on gestation Day 9 in group 4 females.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No relevant changes were detected in food consumption between treated and control females
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
No toxicological relevant changes were seen in uterus weight in treated groups compared to controls.
Gross pathological findings:
no effects observed
Description (incidence and severity):
No macroscopic changes were observed at post mortem examination in treated animals, when compared to the controls. The only macroscopic change, consisting of scab on dorsal neck, is suggested to be incidental.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined

Maternal developmental toxicity

Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
not examined
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not examined
Changes in number of pregnant:
no effects observed
Description (incidence and severity):
Four females in the low dose group and one each in the mid- and high dose groups were found not pregnant at necropsy. Since the mating occurred
before the start of treatment, the incidence of non pregnant females is unrelated to treatment.
One female in each control, low and mid-dose groups had unilateral implantation. The number of females with live foetuses on gestation Day 20 was 24 in the control group, 20 in the low dose and 23 in the mid- and high dose groups.

Effect levels (maternal animals)

Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Basis for effect level:
other: no adverse effects observed up to the limit dose

Maternal abnormalities

Abnormalities:
no effects observed

Results (fetuses)

Fetal body weight changes:
no effects observed
Description (incidence and severity):
A total of 5 small foetuses (< 2.7 g) were detected; 1 out of 317 in the control, 2 out of 280 in the low dose, 1 out of 304 in the mid-dose group and 1 out of 322 in the high dose group.
Since the number of litters affected was similar in the control and treated groups, the finding was considered incidental. No other abnormalities were detected at the external examination of foetuses.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not examined
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
not examined
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Description (incidence and severity):
The skeletal findings did not give rise to concern. Anomalies (AN) and variations (VA) were observed, but they were distributed in a similar way in dose groups and control. Therefore they were considered not treatment related. The only malformation (Forepaw Flexure) was limited to control (one fouetus) and low dose group (3 foetuses/2 litters) and, hence, not dose related. Due to the low incidence and the presence only in the low dose group, this finding was considered incidental.
Visceral malformations:
no effects observed
Description (incidence and severity):
The visceral examination does not give rise to concern. Anomalies (AN) and variations (VA) were observed, but their incidence was similar in the treated groups and the control, and in some cases even more pronounced in the control.
Extreme pelvic dilatation (malformation) was seen in three foetuses/three litters in controls and in one foetus each in the low and high dose group. Due to the lower incidence observed in treated groups respect to controls, this change was considered not related to treatment.
Ureters extremely enlarged and/or kinked were seen in treated and control groups, with similar or lower incidence respect to controls and therefore this finding was considered not treatment-related.
A pulmonary cyst was reported for one foetus in the mid-dose group. Due to the lowincidence and lack of dose-response relationship, this change was considered not related to treatment.

Effect levels (fetuses)

Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: no adverse effects observed up to the limit dose

Fetal abnormalities

Abnormalities:
no effects observed

Overall developmental toxicity

Developmental effects observed:
no

Any other information on results incl. tables

FATE OF FEMALES

 

0 mg/kg bw/d

100 mg/kg bw/d

300 mg/kg bw/d

1000 mg/kg bw/d

Initial group size

24

24

24

24

Not pregnant

0

4

1

1

Unilateral implantation

1

1

1

0

With live foetuses at gestation Day 20

24

20

23

23

 

BODY WEIGHT (g) OF FEMALES WITH LIVE FOETUSES ON GESTATION DAY 20 – GROUP MEAN DATA

Dose level

[mg/kg bw/d]

 

days

 

 

 

 

 

 

0

6

9

12

15

20

0

(n)

Mean

SD

24

234.86

8.55

24

264.87

8.56

24

274.20

10.33

24

288.52

9.93

24

306.45

12.43

24

379.71

21.44

10

(n)

Mean

SD

20

235.79

8.03

20

265.86

8.59

20

274.87

9.07

20

289.63

9.71

20

307.29

12.52

20

381.31

27.30

300

(n)

Mean

SD

23

235.98

11.70

23

266.94

12.76

23

277.94

12.98

23

291.47

13.02

23

310.26

15.29

23

383.07

23.35

1000

(n)

Mean

SD

23

236.99

10.17

23

263.97

8.70

23

276.26

9.53

23

290.95

10.77

23

311.93

10.59

23

389.81

16.25

 

BODY WEIGHT GAIN PER DAY (g) OF FEMALES WITH LIVE FOETUSES ON GESTATION DAY 20 – GROUP MEAN DATA (mean daily body weight gain over the previous period starting from gestation day 0)

Dose level

[mg/kg bw/d]

 

days

 

 

 

 

 

6

9

12

15

20

0

(n)

Mean

SD

24

5.001

0.771

24

3.112

1.349

24

4.772

1.203

24

5.977

2.077

24

14.652

2.805

10

(n)

Mean

SD

20

5.013

0.654

20

3.002

1.548

20

4.920

1.519

20

5.887

2.021

20

14.804

3.328

300

(n)

Mean

SD

23

5.159

1.125

23

3.668

1.036

23

4.508

1.350

23

6.266

1.505

23

14.561

2.275

1000

(n)

Mean

SD

23

4.497

1.237

23

4.098*

1.379

23

4.895

1.469

23

6.994

1.670

23

15.576

1.796

 

* = mean value of group is significantly different from control at p < 0.05

LITTER DATA AND SEX RATIOS - GROUP MEAN DATA

Dose level

[mg/kg bw/d]

 

Corpora lutea

Implan-tations

Uterine deaths

Viable young

% males

Implantation loss [%]

Litter weight [g]

Mean foetal weight [g]

Early

Late

Total

Total

M

F

Pre

Post

Total

0

Mean

SD

(n)

13.88

3.04

24

13.83

3.03

24

 

0.63

1.01

24

0.00

0.00

24

0.63

1.01

24

13.21

3.20

24

6.65

2.35

23

6.83

2.01

24

47.72

14.08

23

0.28

1.37

24

5.44

9.24

24

5.71

9.16

24

47.25

12.06

24

3.58

0.23

24

10

Mean

SD

(n)

14.40

3.70

20

14.20

3.99

20

0.20

0.70

20

0.00

0.00

20

0.20

0.70

20

14.0

4.14

20

6.63

2.27.

19

8.11

1.70

19

45.89

9.49

18

3.06

8.73

20

3.44

11.74

20

5.66

15.69

20

49.62

14.38

20

3.60

0.29

20

300

Mean

SD

(n)

13.61

2.50

23

13.57

2.52

23

0.35

0.57

23

0.00

0.00

23

0.35

0.57

23

13.22

2.33

23

6.65

2.69

23

6.57

2.45

23

50.07

16.57

23

0.33

1.61

23

2.33

3.82

23

2.64

4.54

23

47.26

8.19

23

3.59

0.21

23

1000

Mean

SD

(n)

14.74

2.03

23

14.61

1.90

23

0.57

0.79

23

0.04

0.21

23

0.61

0.84

23

14.00

2.07

23

7.04

2.02

23

6.96

2.23

23

50.35

14.50

23

0.78

2.06

23

4.23

6.01

23

4.96

6.35

23

52.02

7.03

23

3.73

0.25

23

 

EXTERNAL EXAMINATION OF FOETUSES – GROUP INCIDENCE

Dose level

[mg/kg bw/d]

Organ

Cat

Observation

No. foetuses

No. litters

Observed

Affected

%

Observed

Affected

%

0

Whole foetus

 

No abnormalities detected

317

316

99.68

24

-

-

 

Whole foetus

AN

small

317

1

0.32

24

1

4.17

10

Whole foetus

 

No abnormalities detected

280

278

99.29

20

-

-

 

Whole foetus

AN

small

280

2

0.71

20

2

10.00

300

Whole foetus

 

No abnormalities detected

304

303

99.67

23

-

-

 

Whole foetus

AN

small

304

1

0.33

23

1

4.35

1000

Whole foetus

 

No abnormalities detected

322

321

99.69

23

-

-

 

Whole foetus

AN

small

322

1

0.31

23

1

4.35

 

SKELETAL EXAMINATION OF FOETUSES – GROUP INCIDENCE

Dose level

[mg/kg bw/d]

Organ

Cat

Observation

No. foetuses

No. litters

Observed

Affected

%

Observed

Affected

%

0

Forepaw(s)

AN

Metacarpal(s) no ossification 4th

164

62

37.80

24

17

70.83

 

Forepaw(s)

MA

Flexure

164

1

0.61

24

1

4.17

 

Hindpaw(s)

AN

Metatarsal(s) no ossification 4th

164

1

0.61

24

1

4.17

 

Hindpaw(s)

MA

Metatarsal(s) incomplete ossification 4th

164

4

2.44

24

4

16.67

 

Lumbar vertebrae

AN

Centrum dumb-bell shaped

164

1

0.61

24

1

4.17

 

Lumbar vertebrae

AN

Asymmetrical dumb-bell shaped

164

1

0.61

24

1

4.17

 

Pelvic girdle

AN

Pubis incomplete ossification

164

4

2.44

24

3

12.50

 

Pelvic girdle

AN

Ischium incomplete ossification

164

2

1.22

24

2

8.33

 

Ribs

VA

rudimentary 14th

164

32

19.51

24

16

66.67

 

Ribs

VA

short 14th

164

1

0.61

24

1

4.17

 

Sacral vertebrae

AN

Arch(es) incomplete ossification

164

2

1.22

24

2

8.33

 

Skull

AN

Temporal incomplete ossification

164

27

16.46

24

18

75.00

 

Skull

VA

Interparietal incomplete ossification

164

1

0.61

24

1

4.17

 

Skull

VA

Supraoccipital incomplete ossification

164

5

3.05

24

5

20.83

 

Sternebrae

AN

Asymmetrical ossification

164

6

3.66

24

6

25.00

 

Sternebrae

AN

Asymmetrical ossification 5th

164

8

4.88

24

5

20.83

 

Sternebrae

AN

No ossification 6th

164

6

3.66

24

4

16.67

 

Sternebrae

AN

Dumb-bell shaped 5th

164

1

0.61

24

1

4.17

 

Sternebrae

VA

Incomplete ossification 5th

164

41

25.00

24

16

66.67

 

Sternebrae

VA

Incomplete ossification 6th

164

56

34.15

24

13

54.17

 

Sternebrae

VA

No ossification 5th

164

9

5.49

24

6

25.00

 

Sternebrae

VA

Incomplete ossification

164

20

12.20

24

10

41.67

 

Thoracic vertebrae

AN

Centrum no ossification

164

2

1.22

24

2

8.33

 

Thoracic vertebrae

AN

Centrum bipartite

164

1

0.61

24

1

4.17

 

Thoracic vertebrae

VA

Centrum incomplete ossification

164

20

12.20

24

13

54.17

 

Thoracic vertebrae

VA

Centrum dumb-bell shaped

164

11

6.71

24

8

33.33

 

Thoracic vertebrae

VA

Centrum asymmetrical dumb-bell shaped

164

4

2.44

24

4

16.67

100

Forepaw(s)

AN

Metacarpal(s) no ossification 4th

145

53

36.55

20

13

65.00

 

Forepaw(s)

MA

Flexure

145

3

2.07

20

2

10.00

 

Lumbar vertebrae

AN

Centrum dumb-bell shaped

145

1

0.69

20

1

5.00

 

Ribs

VA

14 ribs

145

1

0.69

20

1

5.00

 

Ribs

VA

rudimentary 14th

145

21

14.48

20

14

70.00

 

Ribs

VA

short 14th

145

1

0.69

20

1

5.00

 

Skull

AN

Temporal incomplete ossification

145

25

17.24

20

15

75.00

 

Skull

VA

Supraoccipital incomplete ossification

145

2

1.38

20

2

10.0

 

Skull

VA

Interparietal incomplete ossification

145

1

0.69

20

1

5.00

 

Sternebrae

AN

No ossification 6th

145

2

1.38

20

2

10.0

 

Sternebrae

AN

Asymmetrical ossification

145

4

2.76

20

3

15.00

 

Sternebrae

AN

Asymmetrical ossification 5th

145

5

3.45

20

4

20.00

 

Sternebrae

VA

Incomplete ossification 5th

145

35

24.14

20

15

75.00

 

Sternebrae

VA

No ossification 5th

145

4

2.76

20

4

20.00

 

Sternebrae

VA

Incomplete ossification

145

13

8.97

20

11

55.00

 

Sternebrae

VA

Incomplete ossification 6th

145

52

35.86

20

15

75.00

 

Thoracic vertebrae

AN

Centrum bipartite

145

1

0.69

20

1

5.00

 

Thoracic vertebrae

AN

Centrum no ossification

145

1

0.69

20

1

5.00

 

Thoracic vertebrae

VA

Centrum asymmetrical dumb-bell shaped

145

4

2.76

20

4

20.00

 

Thoracic vertebrae

VA

Centrum dumb-bell shaped

145

27

18.62

20

16

80.00

 

Thoracic vertebrae

VA

Centrum incomplete ossification

145

12

8.28

20

7

35.00

300

Forepaw(s)

AN

Metacarpal(s) no ossification 4th

150

44

29.33

23

15

65.22

 

Lumbar vertebrae

VA

Centrum incomplete ossification

150

2

1.33

23

28.70

 

 

Ribs

AN

Wavy

150

1

0.67

23

1

4.35

 

Ribs

VA

rudimentary 14th

150

21

14.00

23

11

47.83

 

Skull

AN

Temporal incomplete ossification

150

31

20.67

23

13

56.52

 

Skull

VA

Supraoccipital incomplete ossification

150

1

0.67

23

1

4.35

 

Skull

VA

Interparietal incomplete ossification

150

6

4.00

23

4

17.39

 

Sternebrae

AN

Asymmetrical ossification 5th

150

12

8.00

23

8

34.78

 

Sternebrae

AN

No ossification 6th

150

1

0.67

23

1

4.35

 

Sternebrae

AN

Asymmetrical ossification

150

8

5.33

23

7

30.43

 

Sternebrae

VA

Incomplete ossification

150

9

6.00

23

5

21.74

 

Sternebrae

VA

No ossification 5th

150

3

2.00

23

3

13.04

 

Sternebrae

VA

Incomplete ossification 6th

150

36

24.00

23

17

73.91

 

Sternebrae

VA

Incomplete ossification 5th

150

25

16.67

23

13

56.52

 

Thoracic vertebrae

AN

Centrum bipartite

150

1

0.67

23

1

4.35

 

Thoracic vertebrae

VA

Centrum incomplete ossification

150

8

5.33

23

6

26.09

 

Thoracic vertebrae

VA

Centrum asymmetrical dumb-bell shaped

150

1

0.67

23

1

4.35

 

Thoracic vertebrae

VA

Centrum dumb-bell shaped

150

27

18.00

23

15

65.22

1000

Forepaw(s)

AN

Metacarpal(s) no ossification 4th

164

52

31.71

23

15

65.22

 

Hindpaw(s)

VA

Metatarsal(s) incomplete ossification 4th

164

5

3.05

23

3

13.04

 

Pelvic girdle

AN

Pubis incomplete ossification

164

1

0.61

23

1

4.35

 

Pelvic girdle

AN

Ischiumincomplete ossification

164

1

0.61

23

1

4.35

 

Ribs

VA

Rudimentary 14th

164

20

12.20

23

11

47.83

 

Ribs

VA

14 ribs

164

2

1.22

23

1

4.35

 

Ribs

VA

Short 14th

164

2

1.22

23

2

8.70

 

Skull

AN

Temporal incomplete ossification

164

32

19.51

23

16

69.57

 

Skull

VA

Parietalincomplete ossification

164

2

1.22

23

1

4.35

 

Skull

VA

Supraoccipital incomplete ossification

164

4

2.44

23

3

13.04

 

Skull

VA

Interparietal incomplete ossification

164

4

2.44

23

3

13.04

 

Sternebrae

AN

Asymmetrical ossification

164

3

1.83

23

2

8.70

 

Sternebrae

AN

Asymmetrical ossification 5th

164

8

4.88

23

6

26.09

 

Sternebrae

AN

No ossification

164

1

0.61

23

1

4.35

 

Sternebrae

VA

Incomplete ossification 5th

164

30

18.29

23

14

60.87

 

Sternebrae

VA

Incomplete ossification

164

8

4.88

23

5

21.74

 

Sternebrae

VA

Incomplete ossification 6th

164

33

20.12

23

13

56.52

 

Sternebrae

VA

No ossification 5th

164

2

1.22

23

2

8.70

 

Thoracic vertebrae

AN

Centrum no ossification

164

1

0.61

23

1

4.35

 

Thoracic vertebrae

VA

Centrum dumb-bell shaped

164

14

8.54

23

11

47.83

 

Thoracic vertebrae

VA

Centrum incomplete ossification

164

11

6.71

23

8

34.78

 

Thoracic vertebrae

VA

Centrum asymmetrical dumb-bell shaped

164

6

3.66

23

6

26.09

 

VISCERAL EXAMINATION OF FOETUSES – GROUP INCIDENCE

Dose level

[mg/kg bw/d]

Organ

Cat

Observation

No. foetuses

No. litters

Observed

Affected

%

Observed

Affected

%

0

Abdomen

VA

Haemorrhagic

153

23

15.03

24

8

33.33

 

Great vessels

AN

Innominate artery absent

153

1

0.65

24

1

4.17

 

Great vessels

VA

Innominate artery short

153

7

4.58

24

5

20.83

 

Heart

AN

Pericardial haemorrhage

153

23

15.03

24

11

45.83

 

Heart

AN

Pericardial fluid

153

10

6.54

24

6

25.00

 

Heart

AN

Atrium enlarged / slight

153

2

1.31

24

2

8.33

 

Heart

VA

Ventricle enlarged / slight

153

8

5.23

24

6

25.00

 

Kidneys

AN

Ectopic

153

16

10.46

24

10

41.67

 

Kidneys

AN

Pelvic dilatation / moderate

153

5

3.27

24

5

20.83

 

Kidneys

AN

Haemorrhagic

153

1

0.65

24

1

4.17

 

Kidneys

MA

Pelvic dilatation /extreme

153

3

1.96

24

3

12.50

 

Kidneys

VA

Pelvic dilatation / slight

153

6

3.92

24

5

20.83

 

Liver

AN

Mottled

153

6

3.92

24

2

8.33

 

Testis

AN

Displaced

153

5

3.27

24

4

16.67

 

Thoracic cavity

AN

Haemorrhage

153

4

2.61

24

3

12.50

 

Ureter

AN

Enlarged/ moderate

153

9

5.88

24

8

33.33

 

Ureter

AN

Kinked/ moderate

153

1

0.65

24

1

4.17

 

Ureter

MA

Kinked/extreme

153

2

1.31

24

2

8.33

 

Ureter

MA

Enlarged/ extreme

153

4

2.61

24

4

16.67

 

Ureter

VA

Enlarged/ slight

153

19

12.42

24

12

50.00

 

Ureter

VA

Kinked/ slight

153

3

1.96

24

2

8.33

100

Abdomen

VA

Haemorrhagic

135

8

5.93

19

4

21.05

 

Great vessels

VA

Innominate artery short

135

1

0.74

19

1

5.26

 

Great vessels

VA

Innominate artery longer

135

1

0.74

19

1

5.26

 

Heart

AN

Pericardial fluid

135

8

5.93

19

3

15.79

 

Heart

AN

Pericardial haemorrhage

135

2

1.48

19

1

5.26

 

Heart

VA

Ventricle enlarged / slight

135

3

2.22

19

3

15.79

 

Kidneys

AN

Ectopic

135

8

5.93

19

7

36.84

 

Kidneys

AN

Pelvic dilatation / moderate

135

1

0.74

19

1

5.26

 

Kidneys

MA

Pelvic dilatation /extreme

135

1

0.74

19

1

5.26

 

Kidneys

VA

Pelvic dilatation / slight

135

5

3.70

19

5

26.32

 

Testis

AN

Displaced

135

7

5.19

19

6

31.58

 

Thoracic cavity

AN

Haemorrhage

135

1

0.74

19

1

5.26

 

Ureter

AN

Enlarged/ moderate

135

5

3.70

19

4

21.05

 

Ureter

AN

Kinked/ moderate

135

1

0.74

19

1

5.26

 

Ureter

MA

Enlarged/ extreme

135

2

1.48

19

2

10.53

 

Ureter

VA

Enlarged/ slight

135

23

17.04

19

11

57.89

 

Ureter

VA

Kinked/ slight

135

6

4.44

19

5

26.32

300

Abdomen

VA

Haemorrhagic

147

25

17.01

23

9

39.13

 

Great vessels

VA

Innominate artery longer

147

1

0.68

23

1

4.35

 

Heart

AN

Pericardial haemorrhage

147

16

10.88

23

7

30.43

 

Heart

AN

Atrium enlarged / slight

147

1

0.68

23

1

4.35

 

Heart

VA

Pericardial fluid

147

15

10.20

23

9

39.13

 

Heart

VA

Ventricle enlarged / slight

147

7

4.76

23

7

30.43

 

Kidneys

AN

Pelvic dilatation / moderate

147

4

2.72

23

4

17.39

 

Kidneys

AN

Ectopic

147

13

8.84

23

9

39.13

 

Kidneys

VA

Pelvic dilatation / slight

147

5

3.40

23

4

17.39

 

Lungs

AN

Lobe abnormal shape

147

1

0.68

23

1

4.35

 

Lungs

MA

Pulmonary cyst

147

1

0.68

23

1

4.35

 

Testis

AN

Displaced

147

6

4.08

23

5

21.74

 

Ureter

AN

Enlarged/ moderate

147

3

2.04

23

2

8.70

 

Ureter

MA

Enlarged/ extreme

147

2

1.36

23

2

8.70

 

Ureter

MA

Kinked / extreme

147

2

1.36

23

2

8.70

 

Ureter

VA

Kinked/ slight

147

7

4.76

23

6

26.09

 

Ureter

VA

Enlarged/ slight

147

16

10.88

23

9

39.13

1000

Abdomen

VA

Haemorrhagic

158

10

6.33

23

7

30.43

 

Great vessels

VA

Innominate artery short

158

4

2.53

23

3

13.04

 

Great vessels

VA

Innominate artery longer

158

1

0.63

23

1

4.35

 

Heart

AN

Atrium enlarged / slight

158

1

0.63

23

1

4.35

 

Heart

AN

Pericardial haemorrhage

158

22

13.92

23

11

47.83

 

Heart

AN

Pericardial fluid

158

7

4.43

23

4

17.39

 

Heart

VA

Ventricle enlarged / slight

158

3

1.90

23

2

8.70

 

Kidneys

AN

Haemorrhagic

158

1

0.63

23

1

4.35

 

Kidneys

AN

Ectopic

158

13

8.23

23

10

43.48

 

Kidneys

AN

Pelvic dilatation / moderate

158

2

1.27

23

2

8.70

 

Kidneys

MA

Pelvic dilatation / extreme

158

1

0.63

23

1

4.35

 

Testis

AN

Displaced

158

2

1.27

23

1

4.35

 

Ureter

AN

Kinked/ moderate

158

1

0.63

23

1

4.35

 

Ureter

AN

Enlarged/ moderate

158

11

6.96

23

6

26.09

 

Ureter

MA

Kinked / extreme

158

2

1.27

23

2

8.70

 

Ureter

MA

Enlarged/ extreme

158

1

0.63

23

1

4.35

 

Ureter

VA

Enlarged/ slight

158

13

8.23

23

9

39.13

 

Ureter

VA

Kinked/ slight

158

7

4.43

23

4

17.39

AN = anomaly

VA = variant

MA = malformation

Applicant's summary and conclusion

Conclusions:
Neither clinical signs nor signs of reaction to treatment were noted in treated females. No significant differences were noted in body weight, food consumption, gravid uterus weight, litter data and macroscopic observation of treated females when compared to controls. No changes were noted at the skeletal and visceral examinations of the foetuses which were considered treatment-related.
On the basis of these results the dosage of 1000 mg/kg/day could be considered the NOAEL in this study.
Executive summary:

In a prenatal developmental toxicity study according to OECD Guideline 414 (2001) Fatty acids, C16-18, reaction products with diethanolamine (98.81% a.i.) was administered to 24 female Sprague-Dawley rats in 0.5% carboxymethyl cellulose by gavage at dose levels of 0, 100, 300, or 1000 mg/kg bw/day from days 6 through 19 of gestation.

Body weight, daily clinical signs and food consumption were recorded during thein vivophase. All females were caesarean-sectioned on Day 20 post coitum and subjected to post mortem examination. The number of corpora lutea, implantations, early and late intrauterine deaths, live and dead foetuses, uterus weight, foetal weight and sex were recorded. All foetuses were examined for external abnormalities. Approximately one half of the foetuses in each litter was examined for fixed-visceral and skeletal abnormalities.

No animals died during the study. Four females in the low dose group and one each in the mid- and high dose groups were found not pregnant at necropsy. One female in control, low and mid-dose groups had unilateral implantation. The number of females with live fetuses on gestation Day 20 was 24 in the control group, 20 in the low dose and 23 in the mid- and high dose groups.

No signs of toxicological significance were noted during the study and no signs of reactions to treatment were observed during the dosing period.

No relevant differences were noted in body weight and body weight gain of females during the study, between control and treated groups.

No relevant changes were detected in food consumption between treated and control females.

No relevant changes were seen in terminal body weight, uterus weight and absolute weight gain, in treated animals compared to controls.

Litter data and sex ratios were not affected by treatment.

No macroscopic changes were observed at post mortem examination in treated animals, when compared to the controls.

A total of 5 small foetuses (<2.7 g) were detected: 1 in each control, mid- and high dose groups and 2 in the low dose group. No other abnormalities were detected at the external examination of foetuses. No changes were noted at the skeletal examination of the foetuses which were considered treatment-related. No changes were noted at the visceral examination of the foetuses which were considered treatment-related.

On the basis of these results the dosage of 1000 mg/kg/day could be considered the NOAEL in this study.