Registration Dossier

Toxicological information

Developmental toxicity / teratogenicity

Currently viewing:

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:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016

Materials and methods

Test guidelineopen allclose all
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)
Remarks:
testing lab.
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): Geraniol Extra
- Physical state: liquid/colorless, clear
- Homogeneity: given
- Analytical purity: 98.4 corr. area%
- Test substance No.: 10/0046-5
- Lot/batch No.: 00056777LO
- Expiration date of the lot/batch: 31 Oct 2016
- 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.
- Storage condition of test material: room temperature, under light exclusion

Test animals

Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH
- Strain: Crl:WI(Han)
- Age at study initiation: about 10-12 weeks
- Weight at study initiation: 145.5–192.0 g
- Housing: individually in Polycarbonate cages type III with wooden gnawing blocks and dust-free wooden bedding
- Diet: Ground Kliba maintenance diet mouse/rat "GLP", Provimi Kliba SA, Kaiseraugst, Switzerland; ad libitum
- Water: tap water ad libitum
- Acclimation period: between the start of the study (beginning of the experimental phase) and the first administration (GD 6)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: The specific amount of test substance was weighed, topped up with corn oil in a calibrated beaker and intensely mixed with a magnetic stirrer until it is dissolved.
Analytical verification of doses or concentrations:
yes
Details on mating procedure:
- Impregnation procedure: purchased timed pregnant
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”.
Duration of treatment / exposure:
GD 6-19
Frequency of treatment:
once daily
Duration of test:
on GD 20, all surviving females were sacrificed
Doses / concentrations
Remarks:
Doses / Concentrations:
30, 100 and 300 mg/kg bw/day
Basis:
nominal conc.
No. of animals per sex per dose:
25
Control animals:
yes, concurrent vehicle
Details on study design:
In a maternal toxicity rangefinding study dose levels of 300 an 1000 mg/kg bw/d were testet. The test substance was administered to pregnant female rats on GD 6 through GD 19. In this study a dose-related decrease of corrected body weight gain was noted, weight gain was approx. 9 and 12% below concurrent control, respectively. In addition increased relative weights of adrenals (116%), kidneys (117%) and liver (131%) were increased above concur-rent control. Thus dose levels of 30, 100 and 300 mg/kg bw/d were considered appropriate for the present definitive study.

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice a day on working days or once a day on Saturday, Sunday or on public holidays (GD 0-20)

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least once a day, or more often when clinical signs of toxicity were elicited (GD 0-20)

BODY WEIGHT: Yes
- Time schedule for examinations: GD 0, 1, 3, 6, 8, 10, 13, 15, 17, 19 and 20

FOOD CONSUMPTION: Yes
- Time schedule for examinations: GD 0-1, 1-3, 3-6, 6-8, 8-10, 10-13, 13-15, 15-17, 17-19 and 19-20

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: gross pathology
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
- Number of dead fetuses: Yes
Fetal examinations:
- Weight of each fetus: Yes
- Sex: Yes
- Weight of placentas: Yes
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
Statistics:
- DUNNETT's test: 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
- FISHER's exact test: Female mortality, females pregnant at terminal sacrifice, number of litters with fetal findings
- WILCOXON test: Proportions of fetuses with malformations, variations and/or unclassified observations in each litter

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
- MORTALITY:
There were no substance-related or spontaneous mortalities in any females of all test groups.

- CLINICAL SYMPTOMS:
All females of the high-dose group (300 mg/kg bw/d), nearly all (24 out of 25) females of the mid-dose group (100 mg/kg bw/d) and two females of the low-dose group (30 mg/kg bw/d) showed transient salivation during major parts of the treatment period. Salivation persisted in the respective animals only for some minutes after daily gavage dosing (i.e. up to 20 minutes) and was initially observed on GD 7.
No further clinical signs or changes of general behavior, which may be attributed to the test substance, were detected.

- FOOD CONSUMPTION:
The mean food consumption of the dams in all test groups was generally comparable to the concurrent control throughout the entire study period.

- BODY WEIGHT:
The mean body weights (BW) and the average body weight gains (BWC) of the low-, mid- and high-dose dams (30, 100, 300 mg/kg bw/d) were in general comparable to the controls throughout the entire study period. This includes the statistically significantly higher BW/BWC values in the low-dose group on GD 19 and 20 (BW), GD 17-19 and GD 6-19 (BWC).

- CORRECTED (NET) BODY WEIGHT GAIN:
The corrected body weight gain of test groups 1-3 (30, 100 and 300 mg/kg bw/d) revealed no difference of any biological relevance to the concurrent control group. Mean carcass weights remained also unaffected by the treatment.

- UTERUS WEIGHT:
No test-substance related findings.

- NECROPSY FINDINGS:
No test-substance related findings.

- REPRODUCTION DATA:
The conception rate was 100% in all test groups (0, 30, 100 and 300 mg/kg bw/d).
No test-substance related findings in conception rate, in the mean numbers of corpora lutea, implantation sites and viable fetuses or the number of resorptions and the value calculated for postimplantation loss.

Effect levels (maternal animals)

open allclose all
Dose descriptor:
NOAEL
Effect level:
300 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: developmental toxicity
Dose descriptor:
NOAEL
Effect level:
300 mg/kg bw/day
Basis for effect level:
other: maternal toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
- SEX DISTRIBUTION:
comparable to the concurrent control fetuses

- WEIGHT OF PLACENTAE:
comparable to the concurrent control group

- WEIGHT OF FETUSES:
no test substance related effects

- FETAL EXTERNAL MALFORMATIONS:
One fetus with multiple external malformations was recorded in test group 2 (100 mg/kg bw/d). The total incidence of external malformations in treated animals did not differ significantly from the concurrent control group and was comparable to the histori-cal control data.

- FETAL EXTERNAL VARIATIONS:
no external variations were recorded

- FETAL EXTERNAL UNCLASSIFIED OBSERVATIONS:
no unclassified external observations were recorded

- FETAL SOFT TISSUE MALFORMATIONS:
One soft tissue malformation was recorded for two control fetuses and one mid-dose fetus (0 and 300 mg/kg bw/d). The overall incidences of soft tissue malformations were comparable to those found in the historical control data.

- FETAL SOFT TISSUE VARIATIONS:
no soft tissue malformations were recorded

- FETAL SOFT TISSUE UNCLASSIFIED OBSERVATIONS:
no unclassified soft tissue observations were recorded

- FETAL SKELETAL MALFORMATIONS:
Two fetuses bearing skeletal malformations were detected in test groups 0 and 2 (0 and 100 mg/kg bw/d) affecting the skull, vertebral column (with or without involving the ribs) and humerus. An association of these malformations to the treatment is not assumed.

- FETAL SKELETAL VARIATIONS:
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 comparable to the historical control data.

- FETAL SKELETAL UNCLASSIFIED CARTILAGE OBSERVATIONS:
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 and did not show any relation to dosing. The incidence of branched rib cartilage was significantly increased in test group 2 (100 mg/kg bw/d). However, this finding showed no dose-dependency and was therefore assessed to be without biological relevance

Effect levels (fetuses)

Key result
Dose descriptor:
NOAEL
Effect level:
300 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effects observed at highest tested dose

Fetal abnormalities

Abnormalities:
not specified

Overall developmental toxicity

Developmental effects observed:
not specified

Applicant's summary and conclusion