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Toxicological information

Toxicity to reproduction

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Administrative data

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
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report date:
2013

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
yes (incl. QA statement)
Limit test:
yes

Test material

Constituent 1
Reference substance name:
Fatty acids, C8-18 and C18-unsatd., reaction products with diethanolamine and propylene oxide
EC Number:
600-026-8
Cas Number:
1000817-22-0
Molecular formula:
Unspecified
IUPAC Name:
Fatty acids, C8-18 and C18-unsatd., reaction products with diethanolamine and propylene oxide
Details on test material:
- Name of test material (as cited in study report): Kerocom FM 38
- Physical state: liquid, yellow, clear
- Analytical purity: 99.2%
- Lot/batch No.: 2011285
- Storage condition of test material: ambient

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories
- Age at study initiation: 10-11 weeks
- Fasting period before study: no
- Housing: During overnight matings, male and female mating partners were housed together in Makrolon type M III cages. Pregnant animals and their litters were housed together until PND 4
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: yes

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


Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
Kerocom FM 38 was applied as a solution. To prepare this solution, the appropriate amount of test substance was weighed out depending on the desired concentration. Then, drinking water was filled up to the desired volume, subsequently released with a magnetic stirrer. The test substance preparations were produced at least once a week. The administration volume was 10 mL/kg body weight.
Details on mating procedure:
- M/F ratio per cage: 1 male and 1 female
- Length of cohabitation: mating occured overnight until sperm in vaginal smear was detected
- Proof of pregnancy:sperm in vaginal smear; referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged: alone
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The stability of the test substance in drinking water was demonstrated over a period of 7 days at room temperature. As the mixtures were stored no longer than this time period, the stability was guaranteed
Duration of treatment / exposure:
2-week pre-mating and mating period in both sexes, at least 3 days post-mating in males, and the entire gestation period as well as 4 days of lactation and two weeks thereafter in females
Frequency of treatment:
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. Furthermore, examinations as mentioned below were performed
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 100, 300, 1000 mg/kg/d
Basis:
actual ingested
No. of animals per sex per dose:
10 M/F
Control animals:
yes, concurrent vehicle
Details on study design:
The oral route was selected since this was proven to be suitable for the detection of a toxicological hazard.
Positive control:
No

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Cage side observations: A cageside examination was conducted at least once daily for any signs of morbidity, pertinent behavioral changes and signs of overt toxicity. Abnormalities and changes were documented daily for each affected animal.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Detailed clinical observations (DCO) were performed in all animals prior to the administration period and thereafter at weekly intervals.

BODY WEIGHT: Yes
- Time schedule for examinations: Body weight was determined before the start of the administration period in order to randomize the animals. During the administration period body weight was determined on study day 0 (start of the administration period) and thereafter once a week at the same time of the day (in the morning).

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
Generally, food consumption was determined once a week for male and female parental animals,

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes / No / No data
Drinking water consumption was monitored by daily visual inspection of the water bottles for any changes in volume.

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, weight gain, physical or behavioural abnormalities

Postmortem examinations (parental animals):
All parental animals were sacrificed by decapitation under isoflurane anesthesia. The exsanguinated animals were necropsied and assessed by gross pathology, special attention being given to the reproductive organs.

HISTOPATHOLOGY / ORGAN WEIGHTS
The following organs or tissues were fixed in 4% buffered formaldehyde solution or in modified Davidson’s solution:
1. All gross lesions
2. Cervix
3. Coagulating glands
4. Epididymides (modified Davidson’s solution)
5. Ovaries (modified Davidson’s solution)
6. Oviducts
7. Prostate gland
8. Seminal vesicles
9. Testes (modified Davidson’s solution)
10.Vagina
11.Uterus
Postmortem examinations (offspring):
All surviving pups (sacrificed on PND 4 under isoflurane anesthesia with CO2), all stillborn pups and those pups, which died ahead of schedule, were examined externally, eviscerated and their organs were assessed macroscopically.
Statistics:
DUNNETT-test (Food consumption)
FISHER'S EXACT-test (mating indices, 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)
WILCOXON-test (Number of mating days)
Reproductive indices:
Were calculated (see below)
Offspring viability indices:
Were calculated (see below)

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Other effects:
not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
effects observed, treatment-related

Details on results (P0)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
Female animal No. 122 of test group 2 (300 mg/kg bw/d) was sacrificed moribund on study day 14 (mating day 1).
No other animals died or were sacrificed moribund during the study period.
Salivation following treatment was observed (with increasing concentrations of the test item).

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
No treatment-related changes in food consumption and body weights were observed.


REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
The male mating index was 100% in test groups 0 (0 mg/kg bw/d; control group) and 1 (100 mg/kg bw/d) and 90% in test groups 2 (300 mg/kg bw/d) and 3 (1000 mg/kg bw/d). The male fertility indices were 100% in test group 0, 90% in groups 1 and 2 and 70% in group 3.
The female mating index calculated after the mating period for F1 litter was 100% for test groups 0 and 1 and 90% for test groups and 3 (1000 mg/kg bw/d). The mean duration until sperm was detected (GD 0) was 3.4, 2.7, 2.7 and 2.9 days in test groups 0-3. The female fertility index was 100% in test groups 0 and 2, 90% in group 1 and 2 and 77.8% in group 3. The gestation index was 100% in all test groups. The postimplantation loss was not significantly changed when test groups 1-3 were compared to the control animals.
The rate of liveborn pups was 99.1% in test group 0 (control group) and 100% in all treatment test groups

ORGAN WEIGHTS (PARENTAL ANIMALS)
All mean absolute and relative weight parameters showed no significant differences when compared to the control group 0.

GROSS PATHOLOGY (PARENTAL ANIMALS)
All macroscopic 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.

HISTOPATHOLOGY (PARENTAL ANIMALS)
No relevant alterations were seen in the genital organs of male and female rats with failing offspring in test group 3 (1000 mg/kg bw/d). All 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.

Effect levels (P0)

open allclose all
Dose descriptor:
NOAEL
Remarks:
systemic
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no relevant effects observed
Dose descriptor:
NOAEL
Remarks:
reproductive performance, fertility and development
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no relevant effects observed

Results: F1 generation

General toxicity (F1)

Clinical signs:
effects observed, treatment-related
Mortality / viability:
mortality observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
effects observed, treatment-related
Histopathological findings:
not examined

Details on results (F1)

VIABILITY (OFFSPRING)
Only one stillborn pup was seen in animal No. 107 of test group 0, so the rate of liveborn pups was 99.1% in test group 0 (control group) and 100 % in all treatment test groups.
The mean number of delivered F1 pups per dam was evenly distributed among the test groups. The number of delivered F1 pups was 109 in test groups 0 (control group), 1 (100 mg/kg bw/d) and 2 (300 mg/kg bw/d) and 70 in test group 3 (1000 mg/kg bw/d).

CLINICAL SIGNS (OFFSPRING)
The surviving F1 pups of any test group did not show adverse clinical signs up to scheduled sacrifice on PND 4.
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

BODY WEIGHT (OFFSPRING)
Mean body weight of male pups in test group 3 (1000 mg/kg bw/d) was significantly increased on PND 1 (+17%) and 4 (+16%). Thus, the body weight of male and female pups was also significantly increased on PND 1 (+16%) and 4 (+17%). These changes were assessed as being incidental and not related to treatment. No differences in body weight change data were observed.


GROSS PATHOLOGY (OFFSPRING)
No test substance-related findings were observed.

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no relevant effects observed

Overall reproductive toxicity

Reproductive effects observed:
not specified

Applicant's summary and conclusion