Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Under the conditions of an OECD 422 combined repeated dose toxicity study with the reproduction/developmental toxicity screening test, the NOAEL for reproductive performance and fertility was 1000 mg/kg bw/d for the F0 parental rats. The NOAEL for developmental toxicity in the F1 offspring was 1000 mg/kg bw/d.

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012-01-10 to 2012-12-06
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted in accordance with OECD/EU guidelines and principles of GLP.
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
(22 Mar 1996)
Deviations:
no
Qualifier:
according to
Guideline:
other: EPA OPPTS 870.3650: Combined Repeated Dose Toxicity Study With the Reproduction/Developmental Toxicity Screening Test (Jul 2000)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH
- Age at study initiation: 11-12 weeks
- Weight at study initiation: M:303.1 - 337.2 g, F 209.3-237.3g
- Fasting period before study: no
- Housing: individually in Makrolon type M III cages, pregnant animals and their litters were housed together
- Diet (ad libitum): ground Kliba maintenance diet mouse/rat “GLP” meal
- Water (ad libitum): drinking water
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature: 20-24°C
- Humidity: 30-70%
- Air changes: 15 times per hour
- Photoperiod: 12 hours light from 6.00 h to 18.00 h and 12 hours darkness from 18.00 h to 6.00 h

IN-LIFE DATES: From: 16 January 2012 To: M: 13 February 2012, F 07 March 2012
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The test substance solutions in corn oil were prepared at the beginning of the administration period and thereafter in intervals, which took into account the analytical results of the stability verification.
For the preparation of the administration solutions the test substance (after melting by 70°C) was weighed in a graduated flask depending on the dose group, topped up with corn oil and subsequently thoroughly mixed by shaking until it was completely dissolved.

VEHICLE
- Justification for use and choice of vehicle: common vehicle in this kind of study
- Concentration in vehicle: 6.25, 12.5, 25 g/100mL
- Amount of vehicle: 4 mL/kg bw
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: from about 16.00 h until 07.00 - 09.00 h of the following morning for a maximum of 2 weeks
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged individually
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analytical verifications of the stability of the test substance in corn oil for a period of 7 days at room temperature were carried out in a comparable batch prior to the start of the study.
Samples of the test substance solutions were sent to the analytical laboratory once at the beginning of the study for verification of the concentrations.

Determination of octadecylvinylether in corn oil
Method: GC, internal standard method
Apparatus: Agilent 6890 plus GC, Empower-Software (Waters)
Sample preparation: test item was dissolved in dichlormethane
GC conditions:
-Column: Optima 1, 100% Dimethylpolysiloxane, Length: 25 m, Internal diameter: 0.25 mm, Film thickness: 0.25 µm,
- Carrier gas: Nitrogen
- Oven temperature: 120°C to 240°C, 8°C/min, 240°C to 350°C, 40°C/min, 310°C, 20 min isothermal
- Injector temperature: 280°C
- Flow rate: 0.7 mL/min
- Detection: FID
- Detector temperature: 320°C
- Injection volume: 1.0 µL
Duration of treatment / exposure:
The treatment lasted up to one day prior to sacrifice (M: Day 29, F: Day 50)
Frequency of treatment:
Once daily
Details on study schedule:
After 2 weeks of premating treatment the F0 animals were mated to produce F1 generation pups. Mating pairs were from the same test group. Mating was discontinued as soon as sperm was detected in the vaginal smear. F0 animals were examined for their reproductive performance including determination of the number of implantation sites and the calculation of postimplantation loss for all F0 females.
The pups were sexed and examined for macroscopically evident changes on PND 0. They were weighed on PND 1 and on PND 4. Their viability was recorded. At necropsy on PND 4, all pups were sacrificed with CO2, under isoflurane anesthesia, and examined macroscopically for external and visceral findings.
Remarks:
Doses / Concentrations:
250 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
500 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
1000 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
10 animals per sex per dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The aim of this study was to gain initial information on the possible effects of the test item on the integrity and performance of the male and female reproductive systems including gonadal function, mating behavior, conception, gestation and parturition. The study should also provide information about the general toxicological profile including target organs and no-observed-adverse-effect-level (NOAEL) after repeated oral administration.
- Rationale for animal assignment: random, according to body weight four days before the beginning of the administration period.
Positive control:
not applicable
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least once daily
- Cage side observations included are any signs of morbidity, pertinent behavioral changes and signs of overt toxicity

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once prior to the first administration and at weekly intervals during the administration period
- Parameters checked in table [No.1] were examined.

BODY WEIGHT: Yes
- Time schedule for examinations: once a week at the same time of the day

FOOD CONSUMPTION: Yes
- once a week for male and female parental animals
- of the F0 females with evidence of sperm was determined on gestation days (GD) 0, 7, 14 and 20
- of F0 females, which gave birth to a litter was determined on PND 1 and 4

WATER CONSUMPTION: No
Oestrous cyclicity (parental animals):
not examined
Sperm parameters (parental animals):
not examined
Litter observations:
PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, clinical observations, body weight, gross necropsy

GROSS EXAMINATION OF DEAD PUPS:
All stillborn pups and all pups that died before PND 4 were examined externally, eviscerated and their organs were assessed macroscopically.
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals 28 days after the beginning of the administration
- Maternal animals: All surviving animals 50 and 51 days after the beginning of the administration. The females were allowed to deliver and rear their pups until day 4 after parturition.

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table No. 2 and 3 were prepared for microscopic examination and weighed, respectively.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring was sacrificed at 4 days of age.
All pups without notable findings or abnormalities were discarded after their macroscopic evaluation. Animals with notable findings or abnormalities were evaluated on a case-by-case basis, depending on the type of finding.
Statistics:
- Food consumption (parental animals), body weight and body weight change: Simultaneous comparison of all dose groups with the control group using the DUNNETT-test (twosided) for the hypothesis of equal means.

- Male and female mating indices, male and female fertility indices, gestation index, females with liveborn pups, females with stillborn pups, females with all
stillborn pups, live birth index, pups stillborn, pups died, pups cannibalized, pups sacrificed moribund, viability index: Pairwise comparison of each dose group with the control group using FISHER'S EXACT test for the hypothesis of equal proportions.

- Number of mating days: Pairwise comparison of the dose group with the control group using the WILCOXON-test (onesided) with Bonferoni-Holm-Adjustment for the hypothesis of equal medians.

- Feces, rearing, grip strength of forelimbs and hindlimbs, landing foot-splay test, motor activity: Non-parametric oneway analysis using KRUSKAL-WALLIS test (two-sided). If the resulting p-value was equal or less than 0.05, a pairwise comparison of each dose group with the control group was performed using WILCOXON-test (twosided) for the equal medians.

- Clinical pathology parameters: Non-parametric one-way analysis using KRUSKAL-WALLIS test (two-sided). If the resulting p-value was equal or less than 0.05, a pairwise comparison of each dose group with the control group was performed using Wilcoxon-test (two-sided) for the equal medians.

- Weight parameters: Non-parametric one-way analysis using KRUSKALWALLIS test (two-sided). If the resulting p-value was equal or less than 0.05, a pairwise comparison of each dose group with the control group was performed using WILCOXON-test (twosided) for the equal medians.

* for p ≤ 0.05
** for p ≤ 0.01
Reproductive indices:
- Male mating index
- Male fertility index
- Female mating index
- Female fertility index
- Gestation index
Offspring viability indices:
- Live birth index
- Postimplantation loss
CLINICAL SIGNS AND MORTALITY
There were no test substance-related or spontaneous mortalities in any of the groups. With one exception no clinical signs or changes of general behavior, which may be attributedt o the test substance, were detected in any male or female F0 generation parental animals during the whole study including gestation and lactation periods. Several male and female animals of all dose groups showed salivation after treatment during pre-mating, mating, gestation or post-mating. This transient salivation for a few minutes immediately after treatment was likely to be induced by the unpleasant taste of the test substance or by local irritation of the upper digestive tract. It is not considered to be a sign of systemic toxicity.

BODY WEIGHT AND WEIGHT GAIN
Mean body weights and mean body weight change of the male and female F0 generation parental animals in all test substance-treated groups (test groups 1 - 3) showed no decrease in comparison to the concurrent control group during the entire study period.

FOOD CONSUMPTION
Food consumption of the male and female F0 generation parental animals in all test substance-treated groups (test groups 1 - 3; 250, 500 and 1000 mg/kg bw/d) was comparable to the concurrent control group during the entire study period, with one exception. Regarding the statistically significantly increased food consumption of females (test group 3) during premating it was not likely but it could not be excluded that this finding was treatment related. However, this finding was not considered to be adverse.

ORGAN WEIGHTS
The decreased weights of the adrenal gland in female test group 1 (250 mg/kg bw/day) and 3 (1000 mg/kg bw/day) animals and the increase in relative weight of the epididymides of male test group 1 (250 mg/kg bw/day) animals did not show a dose-response relationship and was considered to be incidental.
Although the increase of the relative liver weight in female animals of test group 2 was not statistically significant, there was an increase in absolute weights in this group and the change in liver weights in female animals was dose-dependent. Therefore, the increase in liver weights of male and female animals of test groups 2 and 3 (500 and 1000 mg/kg bw/day) was considered to be treatment-related.
All other mean relative weight parameters did not show significant differences when compared to the control group 0.

GROSS PATHOLOGY
Treatment-related findings consisting of discoloration and prominent acinar pattern were seen in the liver. Light-brown discoloration was noted in 1/10 male test group 2 (500 mg/kg bw/day) and 2/10 male and 6/10 female test group 3 (1000 mg/kg bw/day) animals. Prominent acinar pattern was observed in one female animal of each test group 2 and 3.

HISTOPATHOLOGY: NON-NEOPLASTIC
Liver
Treatment-related fatty change of hepatocytes characterized by vacuoles of varying size was noted in many male and female animals of test groups 2 and 3 (500 and 1000 mg/kg bw/day, respectively) correlating partly to “light-brown discoloration” observed by gross examination. The distribution was different between the sexes: male animals showed centrilobular fatty change while female animals showed a differing pattern with dose: in test group 2, they showed a centrilobular or peripheral pattern whereas in test group 3, there was a peripheral or diffuse change.
Additionally, treatment-related minimal centrilobular hepatocellular hypertrophy was noted in 3/10 male test group 2 (500 mg/kg bw/day) and 4/10 male test group 3 (1000 mg/kg bw/day) animals.
The fatty change in test groups 2 and 3 (both sexes) as well as the centrilobular hypertrophy in test group 3 (male animals) were considered to have contributed to the increased liver weights in these groups.
Tigroid basophilic foci were observed in 4/10 female animals of test group 1 (250 mg/kg bw/day). They occurred singly and were very small, furthermore, there were none in the other treated test groups, and therefore they were considered to be incidental.
All other findings occurred either individually or were biologically equally distributed over control and treatment groups. They were considered to be incidental or spontaneous in origin and without any relation to treatment.

REPRODUCTIVE PERFORMANCE
Male reproduction data:
For all F0 parental males, which were placed with females to generate F1 pups, copulation was confirmed. Thus, the male mating index was 100% in all groups including the controls.
Fertility was proven for most of the F0 parental males within the scheduled mating interval for F1 litter.
One high-dose male, one mid-dose male and two low-dose males did not generate implants in the mated females. Thus, the male fertility index ranged between 80% and 100% without showing any relation to dosing. This reflects the normal range of biological variation inherent in the strain of rats used for this study. The apparently infertile male rats of test groups 1-3 did not show relevant gross lesions.
Female reproduction and delivery data:
The female mating index calculated after the mating period for F1 litter was 100% in all test groups. The mean duration until sperm was detected (GD 0) varied between 1.8 and 3.1 days without any relation to dosing.
The fertility index varied between 80% in test group 1, 90% test groups 2 and 3 and 100% in control. These values reflect the normal range of biological variation inherent in the strain of rats used for this study. None of the non-pregnant females had any relevant gross lesions. The mean duration of gestation was similar in all test groups (i.e. between 22.2 and 22.5 days). The gestation index was 100% in all test groups.
Implantation was not affected by the treatment since the mean number of implantation sites was comparable between all test substance-treated groups and the controls, taking normal biological variation into account (12.9 / 10.7 / 12.1 and 10.2 implants/dam in test groups 0-3 (0, 250, 500 and 1000 mg/kg body weight/day)). There were no statistically significant differences in post-implantation loss between the groups (4.9% / 7.7% / 5.6% / 11.9%), and the mean number of F1 pups delivered per dam remained unaffected (12.3 / 9.7 / 11.4 and 9.8 pups/dam at 0, 250, 500 and 1000 mg/kg bw/d). The rate of liveborn pups was also not affected by the test substance, as indicated by live birth indices of 100% (test group 3 and control), 99.1% (test group 2) and 97.9% (test group 1). Moreover, the number of stillborn pups was comparable between the groups.
Dose descriptor:
NOAEL
Effect level:
250 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: liver toxicity
Remarks on result:
other: Generation: P: general systemic toxicity (migrated information)
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effects observed.
Remarks on result:
other: Generation: P: reproductive performance and fertility (migrated information)
LITTER DATA (OFFSPRING)
The viability index indicating pup mortality during lactation (PND 0 - 4) varied between 99.2% (test group 3) and 100% (test group 2, 1 and control) without showing any association to the treatment.
The mean number of delivered F1 pups per dam and the rates of liveborn and stillborn F1 pups were evenly distributed about the groups. The respective values reflect the normal range of biological variation inherent in the strain used in this study.
The sex distribution and sex ratios of live F1 pups on the day of birth and PND 4 did not show substantial differences between the control and the test substance-treated groups; slight differences were regarded to be spontaneous in nature.

CLINICAL SIGNS (OFFSPRING)
There were no test substance-related adverse clinical signs observed in any of the F1 generation pups of the different test groups.

BODY WEIGHT (OFFSPRING)
No test compound-related influence on F1 pup body weights and pup body weight change were noted in all dose groups.
Two female runts were seen in the control. Five male and two female runts were seen in test group 2 (500 mg/kg bw/d). Two female runts were seen in test group 3 (1000 mg/kg bw/d).

GROSS PATHOLOGY (OFFSPRING)
No findings were observed at gross necropsy in any male or female pups of all test groups. One pup of the high dose group could not be assessed because it had been cannibalized.
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effects observed.
Reproductive effects observed:
not specified

Sex ratio of live F1 pups

PND 0

Test group 0

(0 mg/kg bw/d)

Test group 1

(250 mg/kg bw/d)

Test group 2

(500 mg/kg bw/d)

Test group 3

(1000 mg/kg bw/d)

Live males [%]

49.6

46.3

48.7

58.2

Live females [%]

50.4

53.7

51.3

41.8

PND 4

 

 

 

 

Live males [%]

49.6

46.3

48.7

58.8

Live females [%]

50.4

53.7

51.3

51.2

 

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The study was conducted according to OECD guideline and the principles of GLP. Reliable without restrictions.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

1 -(Vinyloxy)octadecane was given daily as an oily solution to groups of 10 male and 10 female Wistar rats (F0 animals) by stomach tube at doses of 250, 500 and 1000 mg/kg body weight/day (mg/kg bw/day). Control animals (10 male and 10 female Wistar rats) were dosed daily with the vehicle only (corn oil). The duration of treatment covered a 2-week pre-mating and a mating period in both sexes, 2 weeks post-mating in males, and the entire gestation period as well as approximately 2 weeks of the lactation period. After 2 weeks of premating treatment the F0 animals were mated to produce F1 generation pups. Mating pairs were from the same test group. Mating was discontinued as soon as sperm was detected in the vaginal smear. F0 animals were examined for their reproductive performance including determination of the number of implantation sites and the calculation of postimplantation loss for all F0 females.

A detailed clinical observation (DCO) was performed in all animals before initial test substance administration and, as a rule, thereafter at weekly intervals. Food consumption of the F0 parents was determined once weekly during premating. In dams food consumption was determined for gestation days 0 - 7, 7 - 14, 14 - 20 and lactation days 1 - 4.

Body weights of F0 parents were determined once a week, in males throughout the study and in females during premating. During gestation and lactation period, F0 females were weighed on gestation days (GD) 0, 7, 14 and 20, on the day of parturition (postnatal day [PND] 0) and on PND 4.

The pups were sexed and examined for macroscopically evident changes on PND 0. They were weighed on PND 1 and on PND 4. Their viability was recorded. At necropsy on PND 4, all pups were sacrificed with CO2, under isoflurane anesthesia, and examined macroscopically for external and visceral findings.

Clinico-chemical and hematological examinations as well as urinalyses were performed in 5 animals per sex and group towards the end of the administration period. At the end of the administration period a functional observational battery was performed and motor activity was measured in 5 parental males and females per group.

All F0 parental animals were sacrificed by decapitation, under isoflurane anesthesia, and were assessed by gross pathology. Weights of selected organs were recorded and a histopathological examination was performed.

 

The following test substance-related adverse effects/findings were noted:

 

Test group 3: 1000 mg/kg bw/day:

F0 parental animals

• Increased cholesterol, total protein and albumin levels in females

• Minimal to severe hepatocellular fatty change in male and female animals (centrilobular in 6/10 male animals, peripheral or diffuse in 7/10 female animals) in combination with centrilobular hepatocellular hypertrophy in 4/10 male animals

F1 pubs:

• No test substance-related adverse findings

 

Test group 2: 500 mg/kg bw/day:

F0 parental animals

• Minimal to moderate hepatocellular fatty change in male and female animals (centrilobular in 4/10 male animals, centrilobular or peripheral in 6/10 female animals) in combination with centrilobular hepatocellular hypertrophy in 3/10 male animals

F1 pubs

• No test substance-related adverse findings

 

Test group 1: 250 mg/kg bw/day:

F0 parental animals

• No test substance-related adverse findings

F1 pubs

• No test substance-related adverse findings

 

Under the conditions of this OECD 422 combined repeated dose toxicity study with the reproduction/developmental toxicity screening test in Wistar rats the following NOAELs were determined:

The NOAEL for general, systemic toxicity was 250 mg/kg bw/d for the F0 females and males based on liver toxicity observed.

The NOAEL for reproductive performance and fertility was 1000 mg/kg bw/d for the F0 parental rats.

The NOAEL for developmental toxicity in the F1 offspring was 1000 mg/kg bw/d.


Short description of key information:
Under the conditions of an OECD 422 combined repeated dose toxicity study with the reproduction/developmental toxicity screening test in Wistar rats the following NOAEL of Octadecylvinylether were determined:
The NOAEL for reproductive performance and fertility was 1000 mg/kg bw/d for the F0 parental rats.
The NOAEL for developmental toxicity in the F1 offspring was 1000 mg/kg bw/d.

Justification for selection of Effect on fertility via oral route:
Only one study available.

Effects on developmental toxicity

Description of key information
NOAEL: 1000 mg/kg bw/d
    
    
    
    
    

Octadecylvinylether was tested for its prenatal developmental toxicity in Wistar rats. The

test substance was administered as an oily solution to groups of 25 time-mated female

Wistar rats by gavage at doses of 100, 300 and 1000 mg/kg body weight/day (mg/kg bw/d)

NONO

on gestation days (GD) 6 through 19. One control group, consisting of 25 females, was

dosed with the vehicle (corn oil) in parallel. A standard dose volume of 4 mL/kg body weight

was used for each test group.

Octadecylvinylether was tested for its prenatal developmental toxicity in Wistar rats. The

test substance was administered as an oily solution to groups of 25 time-mated female

Wistar rats by gavage at doses of 100, 300 and 1000 mg/kg body weight/day (mg/kg bw/d)

on gestation days (GD) 6 through 19. One control group, consisting of 25 females, was

dosed with the vehicle (corn oil) in parallel. A standard dose volume of 4 mL/kg body weight

was used for each test group.

Octadecylvinylether was tested for its prenatal developmental toxicity in Wistar rats. The

test substance was administered as an oily solution to groups of 25 time-mated female

Wistar rats by gavage at doses of 100, 300 and 1000 mg/kg body weight/day (mg/kg bw/d)

Octadecylvinylether was tested for its prenatal developmental toxicity in Wistar rats. The

test substance was administered as an oily solution to groups of 25 time-mated female

Wistar rats by gavage at doses of 100, 300 and 1000 mg/kg body weight/day (mg/kg bw/d)

on gestation days (GD) 6 through 19. One control group, consisting of 25 females, was

dosed with the vehicle (corn oil) in parallel. A standard dose volume of 4 mL/kg body weight

was used for each test group.

on gestation days (GD) 6 through 19. One control group, consisting of 25 females, was

dosed with the vehicle (corn oil) in parallel. A standard dose volume of 4 mL/kg body weight

was used for each test group.

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
08 Apr 2015 - 23 Nov 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Qualifier:
according to
Guideline:
EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)
GLP compliance:
yes (incl. certificate)
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 20150128
- Expiration date of the lot/batch:
- Purity: 85.5 area-%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature.
- Stability under test conditions: The stability of the test substance under storage conditions
over the test period was guaranteed by the sponsor, and the sponsor holds this responsibility.
- Solubility and stability of the test substance in the solvent/vehicle: The oily test substance preparations
were prepared at the beginning of the administration period and thereafter at intervals, which took
into account the period of established stability.



Species:
rat
Strain:
Wistar
Remarks:
The Crl:WI(Han) strain was selected since extensive historical control data is available from the test facility for Wistar rats. This specific strain has been proven to be sensitive to substances with a teratogenic potential.
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH
- Age at study initiation: 10-12 weeks
- Weight at study initiation: 142.5 - 191.3 g
- Fasting period before study:
- Housing: individual
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24°C
- Humidity (%): 30-70%.
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hours light from 6.00 h to 18.00 h and 12 hours darkness from 18.00 h to 6.00 h

I
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The oily test substance preparations were prepared at the beginning of the administration period and thereafter at intervals,
which took into account the period of established stability. The preparations were kept at room temperature.
For the test substance preparations, the test substance was liquefied at a temperature of about 70°C in a drying chamber.
Thereafter, the specific amount of test substance was weighed, topped up with corn oil in a graduated flask and intensely
mixed with a magnetic stirrer until it is dissolved.
During administration, the preparations were kept homogeneous with a magnetic stirrer.


VEHICLE
- Amount of vehicle (if gavage): volume administered each day was 4 ml/kg body weight.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The results of the analysis of the oily test substance preparations confirmed the correctness of the prepared concentrations.
The analytical values of the samples corresponded to the expected values within the limits of the analytical method,
i.e. were always above 90% and below 110% of the nominal concentrations.
Details on mating procedure:
The animals were paired by the breeder (“time-mated”); the day of evidence of mating (= detection
of vaginal plug/sperm) was referred to as GD 0. The animals arrived on the same day (GD 0) at the
experimental laboratory. The following day was designated as “GD 1”.
The animals were acclimated to the laboratory conditions between start of the study (beginning
of the experimental phase) and first administration (GD 6).
Duration of treatment / exposure:
GD 6 to GD 19
Frequency of treatment:
once a day
Duration of test:
On GD 20, the females were sacrificed in a randomized order and examined macroscopically.
The fetuses were removed from the uterus and investigated
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:
25
Control animals:
yes, concurrent vehicle
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- A cage-side examination was conducted at least once daily for any signs of morbidity, pertinent
behavioral changes and/or signs of overt toxicity. If such signs occurred, the animals were examined
several times daily (GD 0-20).
- Mortality: A check was made twice a day on working days or once a day on Saturdays, Sundays or on
public holidays (GD 0-20).

BODY WEIGHT: Yes
- Time schedule for examinations: All animals were weighed on GD 0, 1, 3, 6, 8, 10, 13, 15, 17, 19 and 20. The body weight
change of the animals was calculated based on the obtained results.
- Corrected (net) body weight gain: Furthermore, the corrected body weight gain was calculated after terminal sacrifice (terminal
body weight on GD 20 minus weight of the unopened uterus minus body weight on GD 6).

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes



POST-MORTEM EXAMINATIONS: Yes / No / No data
- Sacrifice on gestation day 20
- Organs examined: uterue, ovaries

Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes / No / No data
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
Fetal examinations:
- External examinations: Yes:
At necropsy each fetus was weighed, sexed, and external tissues and all orifices were examined
macroscopically. The sex was determined by observing the distance between the anus vand the base of the genitalia.
Furthermore, the viability of the fetuses and the condition of placentae, umbilical cords, fetal membranes, and fluids
were examined. The placentas were weighed and their individual weights were recorded.
Thereafter, the fetuses were sacrificed by a subcutaneous injection of pentobarbital.
After these examinations, approximately one half of the fetuses per dam were eviscerated, skinned and fixed in
ethanol; the other half were placed in Harrison’s fluid for fixation.
- Soft tissue examinations: Yes
The fetuses fixed in Harrison’s fluid were examined for any visceral findings according to the
method of BARROW and TAYLOR. After this examination these fetuses were discarded.
- Skeletal examinations: Yes
The skeletons of the fetuses fixed in ethanol were stained according to a modified method of KIMMEL and TRAMMELL.
Thereafter, the skeletons of these fetuses were examined under a stereomicroscope.
Statistics:
Simultaneous comparison of all dose groups with the control group using the DUNNETT-test (two-sided) for the hypothesis of equal means was used for the following parameters:
Food consumption, body weight, body weight change, corrected body weight gain (net maternal body weight change), carcass weight, weight of unopened uterus, number of corpora lutea, number of implantations, number of resorptions, number of live fetuses, proportions of preimplantation loss, proportions of postimplantation loss, proportions of resorptions, proportion of live fetuses in each litter, litter mean fetal body weight, litter mean placental weight

Pairwise comparison of each dose group with the control group using FISHER'S EXACT test (onesided) for the hypothesis of equal proportions was used for the following parameters:
Female mortality, females pregnant at terminal sacrifice, number of litters with fetal findings

Pairwise comparison of each dose group with the control group using the WILCOXON-test (one-sided) for the hypothesis of equal medians was used for the following parameters:
Proportions of fetuses with malformations, variations and/or unclassified observations in each litter.
Indices:
Conception rate, preimplantation loss, postimplantation loss
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Some females (up to 6 out of 25) of the high-dose group (1000 mg/kg bw/d) showed transient salivation
during the treatment period (GD 16-19). Salivation persisted in the respective animals only for some minutes
after daily gavage dosing (i.e. up to 15 minutes) and was initially observed on GD 16.
It is considered to be treatment-related, likely as a result of the bad taste of the test substance/vehicle
preparation or due to local irritation of the upper digestive tract. It is not considered to be a sign of systemic toxicity

No further clinical signs or changes of general behavior, which may be attributed to the test substance,
were detected in any female at dose levels of 100, 300 or 1000 mg/kg bw/d during the entire study period.
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
The mean body weights (BW) and the average body weight gains (BWC) of the low-, midand
high-dose dams (100, 300, 1000 mg/kg bw/d) were in general comparable to the controls
throughout the entire study period. The statistically significantly increased BWC values during
GD 15-17 in test group 2, and for the treatment period (GD 6-19) in test groups 2 and 3 were
assessed as being incidental.
Corrected (net) body weight gain:
Mean carcass weight of test group 2 was statistically significantly increased in comparison to
the control group. However, the increase was small and not dose-related. Therefore, this was
assessed as incidental.
The corrected body weight gain (terminal body weight on GD 20 minus weight of the unopened
uterus minus body weight on GD 6) was statistically significantly higher than the control
in test groups 1-3 (100, 300 and 1000 mg/kg bw/d). As for the carcass weight, the increase
was rather small and had no effect whatsoever on the well-being of the animals. Thus
this apparent effect was not considered to be toxicologically relevant.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
The mean food consumption of the dams in test groups 1, 2 and 3 (100, 300 and 1000 mg/kg
bw/d) was generally comparable to the concurrent control throughout the entire study period.
The statistically significantly increased food consumption values in test group 3 during
GD 13-17 were assessed to be incidental.
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
Uterus weight
The mean gravid uterus weights of the animals of test groups 1-3 (100, 300 and 1000 mg/kg
bw/d) were not influenced by the test substance. The differences between these groups and
the concurrent control groups revealed no dose-dependency and were assessed to be
without biological relevance.
Gross pathological findings:
no effects observed
Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: no adverse effects observed
Fetal body weight changes:
no effects observed
Changes in sex ratio:
no effects observed
External malformations:
no effects observed
Skeletal malformations:
effects observed, non-treatment-related
Description (incidence and severity):
Skeletal malformations were recorded for one fetus each of test groups 0 and 1 (0 and 100 mg/kg bw/d).
The total incidence of skeletal malformations in treated animals did not differ significantly from the control group
and was comparable to the historical control data.
For all test groups, skeletal variations of different bone structures were observed, with or without effects on corresponding
cartilages. The observed skeletal variations were related to several parts of fetal skeleton and appeared without a relation
to dosing. The overall incidences of skeletal variations were covered by the historical control data.
Fetal skeletal unclassified cartilage observations:
Additionally, some isolated cartilage findings without impact on the respective bone structures, which were
designated as unclassified cartilage observations, occurred in all test groups. The observed unclassified cartilage
findings were related to the skull, the sternum and ribs.
The incidence of ‘bipartite processus xiphoideus’ was statistically significantly increased in test group 2.
As a consequence of this occasional increase, the incidence of total fetal skeletal unclassified cartilage
observations was statistically significantly increased in this test group. Since there is no dose-response
relationship and the finding can be found in the historical control data at a higher frequency, an association to
the treatment and a toxicological relevance is not assumed.
Furthermore, the incidence of ‘notched cartilage between basisphenoid and basioccipital’ was statistically significantly
increased in test group 3 (1000 mg/kg bw/d, mean of affected fetuses/litter: 2.6* %). The mean value was well within
the range of the historical control data (mean 2.4 %, range of 0.0 – 10.0 %) and, therefore, the finding was not
assessed as treatment-related and adverse.
Visceral malformations:
effects observed, non-treatment-related
Description (incidence and severity):
One fetus each in test groups 0 and 2 (0 and 300 mg/kg bw/d) had soft tissue malformations. In one case,
these malformations were associated with external malformations. The total incidence of soft tissue
malformations did not differ significantly from the concurrent control group. The findings were covered by the
historical control data and, therefore, not assessed as treatment-related.
Three soft tissue variations were detected, i.e. short innominate, dilated renal pelvis and dilated ureter.
These variations were neither significantly different from the concurrent control nor dose-dependently altered.
Therefore, they were not considered biologically relevant.
Details on embryotoxic / teratogenic effects:
Fetal external malformations
Three fetuses of one control litter (0 mg/kg bw/d) had external malformations. In two cases, these external malformations
were associated with either soft tissue or skeletal malformations. There were no external malformations in any of the treated
groups.
Fetal external unclassified observations:
One unclassified external observation, i.e. placentae fused, was recorded in two fetuses of the mid-dose group
Since the finding was without statistical significance and not related to dose, it was not considered to be test substance-related.

Assessment of all fetal external, soft tissue and skeletal observations
There were noted external, soft tissue and skeletal malformations in the test groups 0-2 (0, 100 and 300 mg/kg bw/d).
Three fetuses were multiple malformed. One female control fetus showed mandibular micrognathia and a cleft palate
(comprising a short mandible and a small palatine bone), while another female fetus had multiple external malformations
affecting the head (i.e. mandibular micrognathia, cleft palate, open eye) combined with a small lung. Furthermore,
one female mid-dose fetus had multiple soft tissue malformations, i.e. situs inversus (thoracic cavity),
fused or absent lung lobes, membraneous ventricular septum defect. No ontogenetic pattern is recognizable
for these individual malformations nor was there any cluster of any of these individual malformations seen in the other
offspring of these test groups. Most of them can be found in the historical control data of the rat strain.
An association of these findings to the treatment is not assumed. One malformation, i.e. misshapen tuberositas deltoidea,
of test group 1 was not related to the dose and can be found in the historical control data. An association of this finding
to the treatment is not assumed.
External variations did not occur in any of the fetuses in this study. Some soft tissue variations and a range of
skeletal variations were noted in all test groups including the controls. None of the incidences showed a relation to
dosing. The skeletal variations are equally distributed about the different test groups, if normal biological variation
is taken into account, and can be found in the historical control data at a comparable frequency.
No unclassified soft tissue observations were recorded for any of the fetuses in this study.
One unclassified external observation occurred in two mid-dose fetuses. This finding was not considered
to be substance-related. A spontaneous origin was assumed for the unclassified skeletal cartilage observations
which were observed in several fetuses of all test groups( 0, 100, 300 and 1000 mg/kg bw/d).
The distribution and type of these findings do not suggest any relation to treatment.
Finally, fetal examinations revealed that there is no effect of the compound on the respective morphological structures
up to the highest dose tested (1000 mg/kg bw/d).
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no adverse effects observed
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
Under the conditions of this prenatal developmental toxicity study, the oral administration of Octadecylvinylether
to pregnant Wistar rats from implantation to one day prior to the expected day of parturition (GD 6-19) at doses
as high as 1000 mg/kg bw/d caused neither evidence of maternal nor developmental toxicity.
In conclusion, the no observed adverse effect level (NOAEL) for maternal and prenatal developmental toxicity is 1000 mg/kg bw/d.
Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The study was conducted according to OECD guideline and the principles of GLP. Reliable without restrictions.
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

In a prenatal developmental toxicity study (OECD 414) the test substance Octadecylvinylether was administered to pregnant Wistar rats daily by gavage from implantation to one day prior to the expected day of parturition (GD 6-19) to evaluate its potential maternal and prenatal

developmental toxicity.

Generally, clinical observations including food consumption and body weight gain revealed no toxicologically relevant difference between the animals receiving 100, 300 or 1000 mg/kg bw/d Octadecylvinylether and controls. Some females (up to 6 out of 25) of the high-dose group (1000 mg/kg bw/d) showed transient

salivation after treatment. This salivation persisted in the respective females for a few minutes immediately after each administration. It is considered to be treatment-related, likely as a result of the bad taste of the test substance/vehicle preparation or due to local irritation of the upper digestive tract. It is not considered to be a sign of systemic toxicity. No differences of toxicological relevance between the control and the treated groups (100, 300 or 1000 mg/kg bw/d) were determined for any reproductive parameters, such as conception rate, mean number of corpora lutea, mean number of implantations, as well as pre- and

postimplantation loss. Similarly, no influence of the test substance on fetal weight and sex distribution of the fetuses was noted at any dose.

Overall, there was no evidence for toxicologically relevant adverse effects of the test substance on fetal morphology at any dose.

In conclusion, the no observed adverse effect level (NOAEL) for maternal and prenatal developmental toxicity is 1000 mg/kg bw/d

Justification for selection of Effect on developmental toxicity: via oral route:
No adverse effects on development were observed in the OECD 422 combined repeated dose toxicity study with the reproduction/developmental toxicity screening test.

Justification for classification or non-classification

Based on the available studies (OECD 422 and OECD 414), 1 -(vinyloxy)octadecane has not to be classified with respect to reproductive toxicity according to Regulation (EC) No 1272/2008 (CLP, GHS).