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
Study period:
11 February 2015 - 26 March 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted according to OECD 414 Guideline and GLP conditions. Fully adequate for assessment.
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in Section 13.
Cross-reference
Reason / purpose:
read-across: supporting information
Reference
Endpoint:
developmental toxicity
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
11 February 2015 - 26 March 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted according to OECD 414 Guideline and GLP conditions. Fully adequate for assessment.
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in Section 13.
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
Deviation did not affect the purpose or integrity of the study.
Qualifier:
according to
Guideline:
EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries Testing guidelines for Toxicology studies, 12 NohSan No 8147, (24 November 2000)
Qualifier:
according to
Guideline:
other: Commission Regulation (EC) No 440/2008 of 30 May 2008 test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
- Test Material: Octadecene
- CAS Number: CAS 27070-58-2
- Physical State/Appearance : Clear colorless liquid
- Molecular Formula: C18 Isomerized Olefin
- Purity: 94.1%
- Batch Number: 747273
- Label: C18 Isomerised Olefin Lot 747273
- Date Received: 3 March 2014
- Storage Conditions: Room temperature, in the dark and under nitrogen (after opening the container)
- Expiry Date: 10 February 2016

Octadecene used in this study was a typical production sample, according to the details included in the boundary composition in IUCLID section 1.2. Octadecene was chosen in the Higher Olefins category testing strategy because it represents a substance with high di-sub content (category range 0.3 – 94%) and high tri-sub content (category range 1 - 65%). Please see the testing strategy attached in section 13 for further details.
Species:
rat
Strain:
other: Sprague-Dawley Crl:CD (SD) IGS BR strain
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Limited, Margate, Kent
- Age at study initiation: not specified
- Weight at study initiation: 180 to 284g.
- Housing: solid-floor polypropylene cages with stainless steel mesh lids furnished with softwood flakes
- Diet (e.g. ad libitum): A pelleted diet (Rodent 2018C Teklad Global Certified Diet, Harlan UK, Oxon, UK) - free access
- Water (e.g. ad libitum): freely available


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 ºC
- Humidity (%): 50 ± 20%
- Air changes (per hr): 15 air changes per hr
- Photoperiod (hrs dark / hrs light): 12 hr light/12 hr dark

Experimental Starting Date: 11 February 2015
Experimental Completion Date: 26 March 2015

Justification for specie selection: the selected species is a readily available rodent species historically used in safety evaluation studies and is acceptable to appropriate regulatory authorities.


IN-LIFE DATES: From 11 February 2015 to 26 March 2015
Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on exposure:
DIET PREPARATION
- Rate of preparation of diet (frequency): Formulations were prepared once.
- Mixing appropriate amounts with (Type of food): The appropriate concentrations were prepared in Arachis oil solutions.
- Storage temperature of food: Stored at approximately +4 °C in the dark.

Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The stability and homogeneity of the test item formulations were previously determined by Harlan Laboratories Ltd., Shardlow, UK Analytical Services (Project Number 41301656 - Octadecene CAS 27070-58-2: Ninety Day Repeated Dose Oral (Gavage) Toxicity Study in Rats). The results indicate that the prepared formulations were within 90-103% of the nominal concentration confirming the accuracy of the formulation procedure.
Details on mating procedure:
Female animals were delivered in two batches prior to Day 3 of gestation. The day that positive evidence of mating was observed was designated Day 0 of gestation.
Duration of treatment / exposure:
From Day 5 to Day 19 of gestation, by gavage
Frequency of treatment:
Daily
Duration of test:
From Day 5 to Day 19 of gestation
Dose / conc.:
100 mg/kg bw/day
Dose / conc.:
300 mg/kg bw/day
Dose / conc.:
1 000 mg/kg bw/day
No. of animals per sex per dose:
24 Females
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Dose levels were selected based on available toxicity data including a dose range finding study (Harlan Laboratories Ltd report number 41403655)
Maternal examinations:
CAGE SIDE OBSERVATIONS:
Once daily during the gestation period. During the dosing period, observations were recorded immediately before and soon after dosing and one hour post dosing.

BODY WEIGHT:
Individual body weights were recorded on Day 3 (prior to dosing) and on Days 5, 6, 7, 8, 11, 14 and 17 of gestation of gestation, including for surviving animals at terminal kill (Day 20).

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
Food consumption was recorded for each surviving individual animal at Day 3, 5, 8, 11, 14, 17 and 20 of gestation.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study):
Daily by visual inspection of the water bottles for any overt changes.

POST-MORTEM EXAMINATIONS: Yes
- Full external and internal examination ( any macroscopic abnormalities were recorded).

Ovaries and uterine content:
Ovaries and uteri examinations:
i) Number of corpora lutea
ii) Number, position and type of intrauterine implantation
iii) Fetal sex
iv) External fetal appearance
v) Fetal weight
vi) Placental weight
vii) Gravid uterus weight
Fetal examinations:
- Fetal external findings
- Visceral findings
- Skeletal findings and skeletal development

- Number of implantations
- Embryofetal survival
- Litter size
- Sex ratio
- Mean fetal litter and placental weights


Statistics:
The following parameters were analyzed statistically, where appropriate, using the test methods outlined below:
Body weight and body weight change (including adjustment for the contribution of the gravid uterus), food consumption, gravid uterus weight, litter data and fetal litter and placental weights.
Data were first analysed using Shapiro Wilk normality test and Bartlett’s test for homogeneity of variance. Where there was no significance, parametric methodology was applied using one way analysis of variance and, if significant, Dunnett’s multiple comparison test. Where statistical significance was observed, non parametric methodology was applied using Kruskal-Wallis nonparametric analysis of variance; and, if significant, pairwise analysis of control values against treated values using the Mann-Whitney ‘U’ test. Due to the preponderance of non-normal distributions for litter/fetal parameters, these data were routinely analyzed using non-parametric methodology.

Fetal morphology parameters, including skeletal or visceral findings were generally analysed by Kruskal- Wallis and, if significant, Mann-Whitney ‘U’ test.

Probability values (p) are presented as follows:
p<0.001 ***
p<0.01 **
p<0.05 *
p≥0.05 (not significant)
Indices:
Percentage pre-implantation loss
Percentage post-implantation loss
Sex ratio
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
No clinical signs of toxicity were detected.

One control female and one female treated with 300 mg/kg bw/day exhibited generalized fur loss. A further animal treated with 300 mg/kg bw/day also exhibited fur staining. In the absence of findings for high dose females, these observations were considered to be incidental and unrelated to treatment.
Mortality:
no mortality observed
Description (incidence):
There were no unscheduled deaths.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Body weight and body weight gain, including adjustment for the contribution of the gravid uterus, were unaffected by treatment at 100, 300 or 1000 mg/kg bw/day.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Food consumption during gestation was unaffected by treatment at 100, 300 or 1000 mg/kg bw/day.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
Daily visual inspection of water bottles did not reveal any overt intergroup differences.
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:
not examined
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Findings observed at macroscopic necropsy did not indicate any effect of treatment. One control female exhibited generalized fur loss. In the absence of similar findings in high dose females this observation was considered to be incidental and unrelated to treatment.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Other effects:
no effects observed
Description (incidence and severity):
The number of implantations, subsequent embryofetal survival and litter size, sex ratio and mean fetal, litter and placental weights on Day 20 of gestation were unaffected by maternal treatment at 100, 300 and 1000 mg/kg bw/day.
Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
No maternal toxicity
Key result
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: Systemic Toxicity
Key result
Abnormalities:
no effects observed
Fetal body weight changes:
no effects observed
Description (incidence and severity):
Mean fetal, litter and placental weights on Day 20 of gestation were unaffected by maternal treatment at 100, 300 and 1000 mg/kg bw/day.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): Mean fetal, litter and placental weights on Day 20 of gestation were unaffected by maternal treatment at 100, 300 and 1000 mg/kg bw/day.
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Description (incidence and severity):
Sex ratio on Day 20 of gestation was unaffected by maternal treatment at 100, 300 and 1000 mg/kg bw/day.
Changes in litter size and weights:
no effects observed
Description (incidence and severity):
Litter size and weights on Day 20 of gestation were unaffected by maternal treatment at 100, 300 and 1000 mg/kg bw/day.
Changes in postnatal survival:
no effects observed
Description (incidence and severity):
Embryofetal survival was unaffected by maternal treatment at 100, 300 and 1000 mg/kg bw/day.
External malformations:
no effects observed
Description (incidence and severity):
Neither the type, incidence or distribution of findings observed during external examination of the fetuses at necropsy on Day 20 of gestation indicated any adverse effect of maternal treatment on fetal development.
Skeletal malformations:
effects observed, treatment-related
Description (incidence and severity):
Neither the type, incidence or distribution of findings observed during external examination of the fetuses at necropsy on Day 20 of gestation and subsequent detailed visceral and skeletal examination indicated any adverse effect of maternal treatment on fetal development.

Intergroup differences for some skeletal parametersoccasionally attained statistical significance but these isolated differences were considered incidental and did not indicate any disturbance of fetal development.
Visceral malformations:
no effects observed
Description (incidence and severity):
Neither the type, incidence or distribution of findings observed during external examination of the fetuses at necropsy on Day 20 of gestation and subsequent detailed visceral and skeletal examination indicated any adverse effect of maternal treatment on fetal development.
Other effects:
no effects observed
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No embryotoxic/teratogenic effects
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: Developmental Toxicity
Key result
Abnormalities:
no effects observed
Key result
Developmental effects observed:
no

Table 2. Summary of Female Performance

Category

Number of Females at Dose Level (mg/kg bw/day)

0 (control)

100

300

1000

Initial Group Size

24

24

24

24

Pregnant

24

24

24

24

 

Table 3. Group Mean Gravid Uterus Weight and Adjusted Body Weight and Body Weight Change Values

Dose Level

(mg/Kg

bw/day)

 

Body Weight (g)

on Days of

Gestation

Body Weight

Change (g)

during Days of

Gestation

Gravid

Uterus

Weight

(g)

Adjusted

Body

Weight (g)

Day 20

Adjusted

Body Weight

(g)

Change

 

5

20

5-20

Day 20

5-20

0

(control)

Mean

255.7

374.5

118.8

83.667

290.8

35.1

Sd

30.7

48.6

21.4

12.753

39.0

12.5

n

24

24

24

24

24

24

 

100

Mean

256.7

381.8

125.0

86.848

294.9

38.2

Sd

22.4

31.2

18.5

11.060

26.7

13.2

n

24

24

24

24

24

24

 

300

Mean

258.8

380.0

121.2

87.820

292.2

33.4

Sd

20.4

24.7

14.1

8.848

22.8

10.6

n

24

24

24

24

24

24

 

1000

Mean

257.8

374.1

116.3

81.928

292.2

34.3

Sd

24.6

39.6

19.9

15.792

31.6

13.0

n

24

24

24

24

24

24

Table 4. Group Mean Litter Data Values

Dose Level

(mg/Kg

bw/day)

 

Number

of

Corpora

Lutea

Number

of

Implants

Number of

Embryonic/Fetal

Deaths

Implantation

Loss

%

Number of Live

Implants

%

Male

Fetuses

Mean

Male

Fetal

Weight

(g)

Mean

Female

Fetal

Weight

(g)

Mean

Fetal

Weight

(g)

Mean

Placental

Weight

(g)

Litter

Weight

(g)

Total

Placental

Weight

(g)

Early

Late

Total

Pre

Post

Male

Female

Total

0

(control)

Mean

13.9

13.5

0.2

0.2

0.4

2.3

2.9

7.3

5.8

13.2

55.3

4.185

3.981

4.092

0.569

53.555

7.478

Sd

2.0

1.9

0.5

0.7

1.1

3.9

8.1

2.0

1.5

2.3

9.7

0.255

0.273

0.253

0.057

7.966

1.392

n

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

 

100

Mean

14.3

14.0

0.2

0.0

0.2

2.4

1.6

6.5

7.3

13.8

47.2

4.122

3.917

4.012

0.559

55.012

7.656

Sd

1.9

1.9

0.5

0.2

0.7

5.4

5.0

2.2

2.1

2.0

13.7

0.218

0.196

0.196

0.048

7.555

1.166

n

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

 

300

Mean

14.7

14.3

0.1

0.0

0.1

2.2

2.1

7.5

6.5

14.0

53.4

4.126

3.909

4.027

0.563

56.340

7.873

Sd

1.4

1.5

0.4

0.0

0.4

4.3

6.7

1.9

2.0

1.5

13.6

0.194

0.181

0.177

0.061

6.343

1.098

n

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

 

1000

Mean

13.9

13.5

0.4

0.0

0.5

3.4

3.6

6.3

6.7

13.0

49.0

4.127

3.925

4.023

0.557

52.280

7.200

Sd

1.7

2.4

0.8

0.2

0.8

11.2

6.6

1.9

2.1

2.6

11.4

0.250

0.214

0.232

0.059

10.888

1.550

n

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

Table 5. Summary Incidence of Fetal Skeletal Findings

Skeletal

Findings

Dose Level (mg/kg bw/day)

0 (control)

100

300

1000

Number of Fetuses (litters) Examined

150 (24)

160 (24)

162 (24)

150 (24)

NF

NL

%†

NF

NL

%†

NF

NL

%†

NF

NL

%†

Skull

 

Hyoid - incomplete ossification

21

13

14.0

22

13

13.8

12

8

7.2

6

6

3.8*

Vertebral Column

 

Number of pre-sacral vertebrae = 25/27

0

0

0.0

0

0

0.0

4

4

2.7*

0

0

0.0

* Significantly different from control group p<0.05

Table 6. Historical Data - Charles River (UK) Limited - Normal Ranges for Pre-Natal Study Skeletal Fetal Findings in the Sprague-Dawley Crl:CD®(SD) IGS BR Rat

Skeletal findings

Range **

Group mean % of fetuses affected

No. of Litters

{fetuses} examined

Hyoid - incomplete ossification

5 (11.5) - 18.4 [4.0]

255 {1605}

 

Number of pre-sacral vertebrae = 25/27

0 (0.4) – 2.7 [0.8]

255 {1605}

** Range = minimum to maximum value ( ) = group mean  [ ] = 1 standard deviation

 

Where the range exceeds the minimum or maximum possible physiological values, the minimum or maximum physiological values are presented.

Data shown are recorded for presentation purposes, therefore it is not always possible to calculate the range exactly from the mean and SD presented.


Conclusions:
Based on the result of the study, the ‘No Observed Effect Level' (NOEL) for the pregnant female and the ‘No Observed Effect Level' (NOEL) for developmental toxicity is considered to be 1000 mg/kg bw/day.
Executive summary:

In a key Guideline (OECD 414) pre-natal developmental toxicity study, the test material (Octadecene; CAS# 27070-58-2) was administered by gavage to three groups each of twenty-four time mated Sprague-Dawley Crl:CD® (SD) IGS BR strain rats, between Days 5 and 19 of gestation inclusive at dose levels 100, 300, and 1000 mg/Kg bw/day. A further group of twenty-four time mated females was exposed to the vehicle only (Arachis Oil BP) to serve as a control over the same treatment period.

 

Clinical signs, body weight change, food and water consumptions were monitored during the study. All females were terminated on Day 20 of gestation and subjected to gross necropsy including examination of the uterine contents. The number of

corpora lutea, number, position and type of implantation, placental weights, fetal weight, sex and external and internal macroscopic appearance were recorded. Half of each litter were examined for detailed skeletal development and the remaining half were subjected to detailed visceral examination.

 

No mortality was observed through the study period and oral administration of the test material to pregnant rats by oral gavage during gestation at dose levels of 100, 300 and 1000 mg/Kg bw/day did not result in any treatment related effects. The No

Observed Effect Level' (NOEL) for the pregnant female was considered to be 1000 mg/Kg bw/day.

 

No treatment-related changes were detected in the offspring parameters measured. The ‘No Observed Effect Level’ (NOEL) for developmental toxicity was therefore considered to be 1000 mg/Kg bw/day.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
Deviation did not affect the purpose or integrity of the study.
Qualifier:
according to
Guideline:
EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries Testing guidelines for Toxicology studies, 12 NohSan No 8147, (24 November 2000)
Qualifier:
according to
Guideline:
other: Commission Regulation (EC) No 440/2008 of 30 May 2008 test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: Clear colorless liquid
Details on test material:
- Test Material: Octadecene
- CAS Number: CAS 27070-58-2
- Physical State/Appearance : Clear colorless liquid
- Molecular Formula: C18 Isomerized Olefin
- Purity: 94.1%
- Batch Number: 747273
- Label: C18 Isomerised Olefin Lot 747273
- Date Received: 3 March 2014
- Storage Conditions: Room temperature, in the dark and under nitrogen (after opening the container)
- Expiry Date: 10 February 2016

Specific details on test material used for the study:
- Test Material: Octadecene
- CAS Number: CAS 27070-58-2
- Physical State/Appearance : Clear colorless liquid
- Molecular Formula: C18 Isomerized Olefin
- Purity: 94.1%
- Batch Number: 747273
- Label: C18 Isomerised Olefin Lot 747273
- Date Received: 3 March 2014
- Storage Conditions: Room temperature, in the dark and under nitrogen (after opening the container)
- Expiry Date: 10 February 2016

Octadecene used in this study was a typical production sample, according to the details included in the boundary composition in IUCLID section 1.2. Octadecene was chosen in the Higher Olefins category testing strategy because it represents a substance with high di-sub content (category range 0.3 – 94%) and high tri-sub content (category range 1 - 65%). Please see the testing strategy attached in section 13 for further details.

Test animals

Species:
rat
Strain:
other: Sprague-Dawley Crl:CD (SD) IGS BR strain
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Limited, Margate, Kent
- Age at study initiation: not specified
- Weight at study initiation: 180 to 284g.
- Housing: solid-floor polypropylene cages with stainless steel mesh lids furnished with softwood flakes
- Diet (e.g. ad libitum): A pelleted diet (Rodent 2018C Teklad Global Certified Diet, Harlan UK, Oxon, UK) - free access
- Water (e.g. ad libitum): freely available


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 ºC
- Humidity (%): 50 ± 20%
- Air changes (per hr): 15 air changes per hr
- Photoperiod (hrs dark / hrs light): 12 hr light/12 hr dark

Experimental Starting Date: 11 February 2015
Experimental Completion Date: 26 March 2015

Justification for specie selection: the selected species is a readily available rodent species historically used in safety evaluation studies and is acceptable to appropriate regulatory authorities.


IN-LIFE DATES: From 11 February 2015 to 26 March 2015

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on exposure:
DIET PREPARATION
- Rate of preparation of diet (frequency): Formulations were prepared once.
- Mixing appropriate amounts with (Type of food): The appropriate concentrations were prepared in Arachis oil solutions.
- Storage temperature of food: Stored at approximately +4 °C in the dark.

Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The stability and homogeneity of the test item formulations were previously determined by Harlan Laboratories Ltd., Shardlow, UK Analytical Services (Project Number 41301656 - Octadecene CAS 27070-58-2: Ninety Day Repeated Dose Oral (Gavage) Toxicity Study in Rats). The results indicate that the prepared formulations were within 90-103% of the nominal concentration confirming the accuracy of the formulation procedure.
Details on mating procedure:
Female animals were delivered in two batches prior to Day 3 of gestation. The day that positive evidence of mating was observed was designated Day 0 of gestation.
Duration of treatment / exposure:
From Day 5 to Day 19 of gestation, by gavage
Frequency of treatment:
Daily
Duration of test:
From Day 5 to Day 19 of gestation
Doses / concentrationsopen allclose all
Dose / conc.:
100 mg/kg bw/day
Dose / conc.:
300 mg/kg bw/day
Dose / conc.:
1 000 mg/kg bw/day
No. of animals per sex per dose:
24 Females
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Dose levels were selected based on available toxicity data including a dose range finding study (Harlan Laboratories Ltd report number 41403655)

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS:
Once daily during the gestation period. During the dosing period, observations were recorded immediately before and soon after dosing and one hour post dosing.

BODY WEIGHT:
Individual body weights were recorded on Day 3 (prior to dosing) and on Days 5, 6, 7, 8, 11, 14 and 17 of gestation of gestation, including for surviving animals at terminal kill (Day 20).

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
Food consumption was recorded for each surviving individual animal at Day 3, 5, 8, 11, 14, 17 and 20 of gestation.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study):
Daily by visual inspection of the water bottles for any overt changes.

POST-MORTEM EXAMINATIONS: Yes
- Full external and internal examination ( any macroscopic abnormalities were recorded).

Ovaries and uterine content:
Ovaries and uteri examinations:
i) Number of corpora lutea
ii) Number, position and type of intrauterine implantation
iii) Fetal sex
iv) External fetal appearance
v) Fetal weight
vi) Placental weight
vii) Gravid uterus weight
Fetal examinations:
- Fetal external findings
- Visceral findings
- Skeletal findings and skeletal development

- Number of implantations
- Embryofetal survival
- Litter size
- Sex ratio
- Mean fetal litter and placental weights


Statistics:
The following parameters were analyzed statistically, where appropriate, using the test methods outlined below:
Body weight and body weight change (including adjustment for the contribution of the gravid uterus), food consumption, gravid uterus weight, litter data and fetal litter and placental weights.
Data were first analysed using Shapiro Wilk normality test and Bartlett’s test for homogeneity of variance. Where there was no significance, parametric methodology was applied using one way analysis of variance and, if significant, Dunnett’s multiple comparison test. Where statistical significance was observed, non parametric methodology was applied using Kruskal-Wallis nonparametric analysis of variance; and, if significant, pairwise analysis of control values against treated values using the Mann-Whitney ‘U’ test. Due to the preponderance of non-normal distributions for litter/fetal parameters, these data were routinely analyzed using non-parametric methodology.

Fetal morphology parameters, including skeletal or visceral findings were generally analysed by Kruskal- Wallis and, if significant, Mann-Whitney ‘U’ test.

Probability values (p) are presented as follows:
p<0.001 ***
p<0.01 **
p<0.05 *
p≥0.05 (not significant)
Indices:
Percentage pre-implantation loss
Percentage post-implantation loss
Sex ratio

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
No clinical signs of toxicity were detected.

One control female and one female treated with 300 mg/kg bw/day exhibited generalized fur loss. A further animal treated with 300 mg/kg bw/day also exhibited fur staining. In the absence of findings for high dose females, these observations were considered to be incidental and unrelated to treatment.
Mortality:
no mortality observed
Description (incidence):
There were no unscheduled deaths.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Body weight and body weight gain, including adjustment for the contribution of the gravid uterus, were unaffected by treatment at 100, 300 or 1000 mg/kg bw/day.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Food consumption during gestation was unaffected by treatment at 100, 300 or 1000 mg/kg bw/day.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
Daily visual inspection of water bottles did not reveal any overt intergroup differences.
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:
not examined
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Findings observed at macroscopic necropsy did not indicate any effect of treatment. One control female exhibited generalized fur loss. In the absence of similar findings in high dose females this observation was considered to be incidental and unrelated to treatment.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined

Maternal developmental toxicity

Other effects:
no effects observed
Description (incidence and severity):
The number of implantations, subsequent embryofetal survival and litter size, sex ratio and mean fetal, litter and placental weights on Day 20 of gestation were unaffected by maternal treatment at 100, 300 and 1000 mg/kg bw/day.
Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
No maternal toxicity

Effect levels (maternal animals)

Key result
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: Systemic Toxicity

Maternal abnormalities

Key result
Abnormalities:
no effects observed

Results (fetuses)

Fetal body weight changes:
no effects observed
Description (incidence and severity):
Mean fetal, litter and placental weights on Day 20 of gestation were unaffected by maternal treatment at 100, 300 and 1000 mg/kg bw/day.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): Mean fetal, litter and placental weights on Day 20 of gestation were unaffected by maternal treatment at 100, 300 and 1000 mg/kg bw/day.
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Description (incidence and severity):
Sex ratio on Day 20 of gestation was unaffected by maternal treatment at 100, 300 and 1000 mg/kg bw/day.
Changes in litter size and weights:
no effects observed
Description (incidence and severity):
Litter size and weights on Day 20 of gestation were unaffected by maternal treatment at 100, 300 and 1000 mg/kg bw/day.
Changes in postnatal survival:
no effects observed
Description (incidence and severity):
Embryofetal survival was unaffected by maternal treatment at 100, 300 and 1000 mg/kg bw/day.
External malformations:
no effects observed
Description (incidence and severity):
Neither the type, incidence or distribution of findings observed during external examination of the fetuses at necropsy on Day 20 of gestation indicated any adverse effect of maternal treatment on fetal development.
Skeletal malformations:
effects observed, treatment-related
Description (incidence and severity):
Neither the type, incidence or distribution of findings observed during external examination of the fetuses at necropsy on Day 20 of gestation and subsequent detailed visceral and skeletal examination indicated any adverse effect of maternal treatment on fetal development.

Intergroup differences for some skeletal parametersoccasionally attained statistical significance but these isolated differences were considered incidental and did not indicate any disturbance of fetal development.
Visceral malformations:
no effects observed
Description (incidence and severity):
Neither the type, incidence or distribution of findings observed during external examination of the fetuses at necropsy on Day 20 of gestation and subsequent detailed visceral and skeletal examination indicated any adverse effect of maternal treatment on fetal development.
Other effects:
no effects observed
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No embryotoxic/teratogenic effects

Effect levels (fetuses)

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: Developmental Toxicity

Fetal abnormalities

Key result
Abnormalities:
no effects observed

Overall developmental toxicity

Key result
Developmental effects observed:
no

Any other information on results incl. tables

Table 2. Summary of Female Performance

Category

Number of Females at Dose Level (mg/kg bw/day)

0 (control)

100

300

1000

Initial Group Size

24

24

24

24

Pregnant

24

24

24

24

 

Table 3. Group Mean Gravid Uterus Weight and Adjusted Body Weight and Body Weight Change Values

Dose Level

(mg/Kg

bw/day)

 

Body Weight (g)

on Days of

Gestation

Body Weight

Change (g)

during Days of

Gestation

Gravid

Uterus

Weight

(g)

Adjusted

Body

Weight (g)

Day 20

Adjusted

Body Weight

(g)

Change

 

5

20

5-20

Day 20

5-20

0

(control)

Mean

255.7

374.5

118.8

83.667

290.8

35.1

Sd

30.7

48.6

21.4

12.753

39.0

12.5

n

24

24

24

24

24

24

 

100

Mean

256.7

381.8

125.0

86.848

294.9

38.2

Sd

22.4

31.2

18.5

11.060

26.7

13.2

n

24

24

24

24

24

24

 

300

Mean

258.8

380.0

121.2

87.820

292.2

33.4

Sd

20.4

24.7

14.1

8.848

22.8

10.6

n

24

24

24

24

24

24

 

1000

Mean

257.8

374.1

116.3

81.928

292.2

34.3

Sd

24.6

39.6

19.9

15.792

31.6

13.0

n

24

24

24

24

24

24

Table 4. Group Mean Litter Data Values

Dose Level

(mg/Kg

bw/day)

 

Number

of

Corpora

Lutea

Number

of

Implants

Number of

Embryonic/Fetal

Deaths

Implantation

Loss

%

Number of Live

Implants

%

Male

Fetuses

Mean

Male

Fetal

Weight

(g)

Mean

Female

Fetal

Weight

(g)

Mean

Fetal

Weight

(g)

Mean

Placental

Weight

(g)

Litter

Weight

(g)

Total

Placental

Weight

(g)

Early

Late

Total

Pre

Post

Male

Female

Total

0

(control)

Mean

13.9

13.5

0.2

0.2

0.4

2.3

2.9

7.3

5.8

13.2

55.3

4.185

3.981

4.092

0.569

53.555

7.478

Sd

2.0

1.9

0.5

0.7

1.1

3.9

8.1

2.0

1.5

2.3

9.7

0.255

0.273

0.253

0.057

7.966

1.392

n

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

 

100

Mean

14.3

14.0

0.2

0.0

0.2

2.4

1.6

6.5

7.3

13.8

47.2

4.122

3.917

4.012

0.559

55.012

7.656

Sd

1.9

1.9

0.5

0.2

0.7

5.4

5.0

2.2

2.1

2.0

13.7

0.218

0.196

0.196

0.048

7.555

1.166

n

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

 

300

Mean

14.7

14.3

0.1

0.0

0.1

2.2

2.1

7.5

6.5

14.0

53.4

4.126

3.909

4.027

0.563

56.340

7.873

Sd

1.4

1.5

0.4

0.0

0.4

4.3

6.7

1.9

2.0

1.5

13.6

0.194

0.181

0.177

0.061

6.343

1.098

n

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

 

1000

Mean

13.9

13.5

0.4

0.0

0.5

3.4

3.6

6.3

6.7

13.0

49.0

4.127

3.925

4.023

0.557

52.280

7.200

Sd

1.7

2.4

0.8

0.2

0.8

11.2

6.6

1.9

2.1

2.6

11.4

0.250

0.214

0.232

0.059

10.888

1.550

n

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

24

Table 5. Summary Incidence of Fetal Skeletal Findings

Skeletal

Findings

Dose Level (mg/kg bw/day)

0 (control)

100

300

1000

Number of Fetuses (litters) Examined

150 (24)

160 (24)

162 (24)

150 (24)

NF

NL

%†

NF

NL

%†

NF

NL

%†

NF

NL

%†

Skull

 

Hyoid - incomplete ossification

21

13

14.0

22

13

13.8

12

8

7.2

6

6

3.8*

Vertebral Column

 

Number of pre-sacral vertebrae = 25/27

0

0

0.0

0

0

0.0

4

4

2.7*

0

0

0.0

* Significantly different from control group p<0.05

Table 6. Historical Data - Charles River (UK) Limited - Normal Ranges for Pre-Natal Study Skeletal Fetal Findings in the Sprague-Dawley Crl:CD®(SD) IGS BR Rat

Skeletal findings

Range **

Group mean % of fetuses affected

No. of Litters

{fetuses} examined

Hyoid - incomplete ossification

5 (11.5) - 18.4 [4.0]

255 {1605}

 

Number of pre-sacral vertebrae = 25/27

0 (0.4) – 2.7 [0.8]

255 {1605}

** Range = minimum to maximum value ( ) = group mean  [ ] = 1 standard deviation

 

Where the range exceeds the minimum or maximum possible physiological values, the minimum or maximum physiological values are presented.

Data shown are recorded for presentation purposes, therefore it is not always possible to calculate the range exactly from the mean and SD presented.


Applicant's summary and conclusion

Conclusions:
Based on the result of the study, the ‘No Observed Effect Level' (NOEL) for the pregnant female and the ‘No Observed Effect Level' (NOEL) for developmental toxicity is considered to be 1000 mg/kg bw/day.
Executive summary:

In a key Guideline (OECD 414) pre-natal developmental toxicity study, the test material (Octadecene; CAS# 27070-58-2) was administered by gavage to three groups each of twenty-four time mated Sprague-Dawley Crl:CD® (SD) IGS BR strain rats, between Days 5 and 19 of gestation inclusive at dose levels 100, 300, and 1000 mg/Kg bw/day. A further group of twenty-four time mated females was exposed to the vehicle only (Arachis Oil BP) to serve as a control over the same treatment period.

 

Clinical signs, body weight change, food and water consumptions were monitored during the study. All females were terminated on Day 20 of gestation and subjected to gross necropsy including examination of the uterine contents. The number of

corpora lutea, number, position and type of implantation, placental weights, fetal weight, sex and external and internal macroscopic appearance were recorded. Half of each litter were examined for detailed skeletal development and the remaining half were subjected to detailed visceral examination.

 

No mortality was observed through the study period and oral administration of the test material to pregnant rats by oral gavage during gestation at dose levels of 100, 300 and 1000 mg/Kg bw/day did not result in any treatment related effects. The No

Observed Effect Level' (NOEL) for the pregnant female was considered to be 1000 mg/Kg bw/day.

 

No treatment-related changes were detected in the offspring parameters measured. The ‘No Observed Effect Level’ (NOEL) for developmental toxicity was therefore considered to be 1000 mg/Kg bw/day.