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Key value for chemical safety assessment

Effects on developmental toxicity

Description of key information

In a two-generation feeding study in male and female Fisher rats (Chin et al., 1994), the test item did not show any adverse effects on the development of the offspring.

In a two-generation developmental toxicity screening study in female ICR/SIM mice (Seidenberg et al., 1986), the test item did not show any adverse effects on the development of the offspring.

Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
- Principle of test: Effect of Conjugated Linoleic Acid (CLA) on rat development and growth.
- Short description of test conditions: The test item was administered to female animals after mating in concentrations of 0.25 g/100 g diet or 0.5 g/100 g diet in the diet. The pups received the test item via mother milk or via food after weaning.
- Parameters analysed / observed: CLA content in rat milk, CLA tissue concentrations in dams and fetusses, weight (gain),
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
other: Fisher
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan, Sprague Dawley, Indianapolis, USA
- Age at study initiation:
Experiment I: 8 weeks
Experiment II: 10 weeks
- Housing: individually
- Diet: nonpurified diet (Ralston Purina, St. Louis, MO, USA)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 22
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
CLA was mixed in corn oil before addition to the diet.

DIET PREPARATION
- Rate of preparation of diet: weekly
- Mixing appropriate amounts with: basal diet
- Storage temperature of food: +4 °C (under nitrogen to prevent autooxidation)

Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
- Impregnation procedure: cohoused
- M/F ratio per cage: 1/1
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy.
Duration of treatment / exposure:
up to 10 weeks
Frequency of treatment:
daily
Dose / conc.:
0 other: g/100 g diet
Remarks:
Control diet (CD)
Dose / conc.:
0.25 other: g/100 g diet
Remarks:
control diet with CLA
Dose / conc.:
0.5 other: g/100 g diet
Remarks:
control diet with CLA
Dose / conc.:
0.5 other: g/100 g diet
Remarks:
CD during gestation, CLA during lacation
Control animals:
yes, plain diet
Maternal examinations:
BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- Time schedule for examinations: weekly

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20, 10 animals of each group
- Organs examined: liver, mammary gland, skeletal muscle and abdominal adipose tissues were then removed. Fetuses were removed, weighed and examined visually for abnormalities.

OTHER:
Collection of individual milk samples at d10 of lactation. Total protein in milk was determined.
Quantification of RNA and DNA from mammary glands was performed.
Statistics:
Data were analysed by ANOVA. Significant differences among means were determined by Tukey's comparisons.
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
not examined
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Number of abortions:
not examined
Pre- and post-implantation loss:
not examined
Total litter losses by resorption:
not examined
Early or late resorptions:
not examined
Dead fetuses:
not examined
Changes in pregnancy duration:
not examined
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not examined
Changes in number of pregnant:
not examined
Other effects:
not examined
Key result
Dose descriptor:
NOAEL
Effect level:
> 0.5 other: g/100 g
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Abnormalities:
no effects observed
Fetal body weight changes:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): effects observed, treatment-related
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): Pups which received CLA during gestation and lactation showed a significant increase of body weight compared to control pups.
Reduction in number of live offspring:
not examined
Changes in sex ratio:
not examined
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
not examined
External malformations:
no effects observed
Skeletal malformations:
not examined
Visceral malformations:
not examined
Other effects:
not examined
Key result
Dose descriptor:
NOAEL
Effect level:
> 0.5 other: g/100 g
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Abnormalities:
no effects observed
Key result
Developmental effects observed:
no
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
- Principle of test: A test system for identifying potential teratogenicity based on growth and viability of embryonic, fetal, and postnatal mice
- Short description of test conditions: A single, minimally toxic dose, expected to result in significant maternal weight reduction, up to 10% mortality, or other clinical signs of overt toxicity, was selected based on previous range-finding studies in nonpregnant female mice. 26 to 30 timed-pregnant mice were treated by oral intubation on days 8 through 12 of gestation (day 1 = day plug found).
Dosages (1 to 1.4 mL/ 100 g of body weight) were based on body weights at day 7 of gestation; mice were also weighed on day 13 and at 1 day postpartum. Mice were allowed to deliver, and neonates were examined, counted, and weighed on the day of birth (day 1) and day 3. Dead neonates were recovered from the nest and externally examined for abnormalities. Dams that had not given birth by gestation day 21 or 22 were necropsied and their uteri were examined.
GLP compliance:
not specified
Limit test:
no
Species:
mouse
Strain:
ICR
Remarks:
ICR/SIM
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Simonsen Laborataries (Gilray, CA)
- Age at study initiation: adult
- Weight at study initiation: 32 - 36 g
- Housing: individually
- Diet: Siniansen Custam Lab Diet 7, ad libitum
- Water: UV-purified drinking water, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
- Impregnation procedure: Mice were received from the supplier timed-pregnant or were bred in-house, due to the low pregnancy rate of the delivered mice.
- Proof of pregnancy: vaginal plug referred to as day 1 of pregnancy
Duration of treatment / exposure:
Day 8 - 12 of gestation
Frequency of treatment:
daily
Duration of test:
5 days
Dose / conc.:
0 other: mL/100 g bw
Dose / conc.:
14 other: mL/kg bw
No. of animals per sex per dose:
30
Control animals:
yes, concurrent no treatment
Details on study design:
- Dose selection rationale: Based an previous range-finding studies in nonpregnant female mice.
Maternal examinations:

BODY WEIGHT: Yes
- Time schedule for examinations: Day 7 and 13 of gestation, Day 1 post partum.

POST-MORTEM EXAMINATIONS: Yes, dams that had not given birth by gestation day 21 or 22 were necropsied and their uteri were examined.

Fetal examinations:
- External examinations and weighing: Yes, all per litter on day 1 and 3
- Other: Dead neonates were recovered from the nest and externally examined for abnormalities
Statistics:
Maternal body weight gain during treatment (day 13 minus day 7 weight) and neonatal body weight: two-tailed analysis of variance
Live and dead litter size data: one-tailed analysis of variance
Neonatal survival ratios: one-tailed Fisher's exact probability test
Indices:
- Average number of viable neonates
Clinical signs:
not specified
Dermal irritation (if dermal study):
not examined
Mortality:
mortality observed, non-treatment-related
Description (incidence):
One animal died during the study.
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Number of abortions:
not specified
Pre- and post-implantation loss:
not specified
Total litter losses by resorption:
no effects observed
Early or late resorptions:
not specified
Dead fetuses:
not specified
Changes in pregnancy duration:
not examined
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Changes in number of pregnant:
not examined
Other effects:
not examined
Key result
Dose descriptor:
NOAEL
Effect level:
> 14 other: mL/kg bw/d
Based on:
test mat.
Remarks on result:
not measured/tested
Key result
Abnormalities:
no effects observed
Fetal body weight changes:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
not examined
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
no effects observed
External malformations:
no effects observed
Skeletal malformations:
not specified
Visceral malformations:
not specified
Other effects:
not examined
Key result
Dose descriptor:
NOAEL
Effect level:
> 14 other: mL/kg bw/d
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Abnormalities:
no effects observed
Key result
Developmental effects observed:
no
Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
Study duration:
subacute
Species:
rat
Quality of whole database:
Well documented publication.
Additional information

In a two-generation feeding study in male and female Fisher rats (Chin et al., 1994), the effect of the test item on fetal and postnatal development of the offsprings was investigated. The test item was administered via feeding in concentrations of 0.25 or 0.5 %. Test item administration in P0 animals started with the confirmation of succesful mating (sperm in vaginal smear). F1 animals received the test item via lactation or via food after weaning, respectively. Pups which received a diet supplemented with 0.5 % test item showed a significantly elevated body weight. However, no adverse effects were observed.

A similar result was observed in a two-generation in vivo developmental toxicity screening study in female ICR/SIM mice (Seidenberg et al., 1986). The effect of the test item on fetal and postnatal development of the offsprings was investigated. The test item was administered via oral gavage in a concentration of 14 mL/kg bw. Test item administration in P0 animals started at day 8 of gestation after the confirmation of succesful mating (vaginal plug = day 1). Test item treatment did not show adverse effects on litter size number or neonatal development.

Toxicity to reproduction: other studies

Description of key information

In an two-generation feeding study in female P0 and F1 Sprague-Dawley rats (Reynolds et al., 2015), the test item did not show any adverse effects.

Link to relevant study records
Reference
Endpoint:
toxicity to reproduction: other studies
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
GLP compliance:
not specified
Species:
rat
Strain:
Sprague-Dawley
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: inhouse breeding
- Housing: individually after confirmation of pregnancy
- Age at study initiation: adult, 110 days
- Diet: standard purified control diet (CD), standard purified control diet with c9,t11-CLA (CLA), high fat diet (HF), HF with c9,t11-CLA (HFCLA)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 25
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
P0 females: 160 days
F1: 150 days
Frequency of treatment:
Daily
Dose / conc.:
0 other: %
Remarks:
control diet (CD)
Dose / conc.:
0 other: %
Remarks:
high fat diet (HF)
Dose / conc.:
1 other: %
Remarks:
control diet with c9,t11-CLA (CLA)
Dose / conc.:
1 other: %
Remarks:
HF with c9,t11-CLA (HFCLA)
No. of animals per sex per dose:
7 dams
8 pups/litter
Control animals:
yes, plain diet
Details on study design:
Female rats were fed for 10 days prior to time mated. Mating point was determined using a estrous cycle monitor (EC-40; Fine Science Tools).
Statistics:
Statistical analysis was performed using SigmaPlot 12.0 (Systat Software Inc.). All data were analyzed by two-way factorial ANOVA, with maternal HF and maternal CLA intake as factors. Holm-Sidak post hoc tests were performed where indicated to detect further differences between groups.
Key result
Sex:
male/female
Remarks on result:
other: no treatment related adverse effects were observed.
The age of pubertal onset was significantly reduced in high fat diet (HF) compared to control diet (CD), control diet with conjugated linoleic acid (CLA), and high fat diet with conjugated linoleic acid (HFCLA) offspring. The weight at puberty was significantly increased in HF offsprings.
HF offspring had significantly increased plasma leptin concentrations compared to CD, CLA, and HFCLA offspring. Triglyceride as well as cholesterol concentrations were significantly increased in HF compared to CD, CLA, and HFCLA fed animals. An significant increase in hepatic IGF-1 expression in HF-offsprings at P24 was observed compared to all other groups, but no significant differences were observed at P150. IL-1b and IL-6 expression was increased in HF-offsprings compared to all other groups. However, there was no difference between groups in TNFa or IL-10 expression. CD36, DGAT1, and FASN were significantly upregulated in offspring of HF mothers compared with CD, CLA, and HFCLA offspring. ApoE was significantly increased in HF and HFCLA compared to CD and CLA groups. Female HF-offsprings showed prolonged or irregular estrous cycles compared to CD, CLA, and HFCLA groups.
Additional information

In an two-generation feeding study in female Sprague-Dawley rats (Reynolds et al., 2015), the effect of the test item on a high fat diet was determined. The test item was administered via feeding in a concentration of 1 %. Two different food preperations were investigated, a control diet (CD), containing 10 % kcal from fat and a high fat diet (HF), containing 45 % kcal from fat. Test item administration in P0 animals started 10 days before mating. F1 animals recieved the test item via lactation or via food after weaning, respectively. No test item related adverse effects were observed.

Justification for classification or non-classification

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. As a result the substance is not considered to be classified as reproductive and developmental toxicant under Regulation (EC) No 1272/2008, as amended for the tenth time in Regulation (EU) No 2017/776.

Additional information