Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Key studies on reproductive toxicity by the oral route are available for the following category members:

Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test (OECD 422), rat: NOAEL oral (fertility) 1000 mg/kg bw/day; CAS# 112-85-6, C22

No hazard for reproductive toxicity was identified for fatty acids.

No data are available for reproductive toxicity after dermal exposure and inhalation, respectively.

Link to relevant study records

Referenceopen allclose all

Endpoint:
screening for reproductive / developmental toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Japan, Hino, Tokyo
- Age at study initiation: 8 weeks
- Weight at study initiation: male: 312.1 - 363.7 g; female: 205.3 - 230.8 g
- Housing: metal wire floor cages
- Diet (ad libitum): CE-2, CLEA Japan
- Water (ad libitum): tap water
- Acclimation period: 7 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 ± 1
- Humidity (%): 50 - 65
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 / 12
Route of administration:
oral: gavage
Vehicle:
olive oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
prepared more frequently than once a week; aliquots were kept by each concentration refrigerated in airtight conditions

VEHICLE
- Justification for use and choice of vehicle (if other than water): due to insolubility in water
- Amount of vehicle (if gavage): 5 mL/kg bw
- Lot/batch no. (if required): V6H2050
Details on mating procedure:
- Impregnation procedure: cohoused
- M/F ratio per cage: 1/1
- Length of cohabitation: up to two weeks
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
- Further mating after two unsuccessful attempts: no
- After successful mating each pregnant female was caged (how): individually
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
- males: 42 days
- females: from 14 days prior to mating to day 3 of lactation
Frequency of treatment:
once daily
Details on study schedule:
- Age at mating of the mated animals in the study: 10 weeks
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
13
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: based on preliminary result in a 14 day-repeated dose toxicity study, where no signs of toxicity were found
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least once a day

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: males: days 1, 8, 15, 22, 29, 36, 42; pregnancy females: premating days 1, 8, 15; pregnancy days: 0, 7, 14, 20; lactation days: 0, 4; non pregnancy females: day 1, 8, 15, 22, 25

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No

HAEMATOLOGY: Yes (males only)
- Time schedule for collection of blood: prior to autopsy
- Anaesthetic used for blood collection: Yes (identity): pentobarbital sodium
- Animals fasted: 18 - 24 hours before sacrifice
- How many animals: all surviving animals
- Parameters checked: red blood cell count (RBC), white blood cell count (WBC), haemoglobin content (Hb), mean corpuscular volume (MCV), ahematocrit (Ht), mean corpuscular haemoglobin content (MCH), mean corpuscular haemoglobin concentration (MCHC)

CLINICAL CHEMISTRY: Yes (males only)
- Animals fasted: 18 - 24 hours before sacrifice
- How many animals: all surviving animals
- Parameters checked: total protein, albumin, total cholesterol concentration, glucose, urea nitrogen, creatinine, alkaline phosphatase activity, GOT, GPT, γ-GTP, triglyceride concentration, inorganic phosphorus, total bilirubin, calcium, sodium, potassium, chlorine, A/G ratio

URINALYSIS: No
Oestrous cyclicity (parental animals):
not examined
Sperm parameters (parental animals):
Parameters examined in all male parental generations:
testis weight, epididymis weight
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was determined for pups born or found dead.
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: all surviving animals
- Maternal animals: all surviving animals

HISTOPATHOLOGY / ORGAN WEIGHTS
Organ weights:
male: heart, liver, kidneys, thymus, testes, epididymides
female: heart, liver, kidneys, thymus

Pathological findings and Histopathology (control and 1000 mg/kg bw/day group):
male: heart, liver, spleen, kidney, adrenal, testis, epididymis, brain, thymus, bladder
female: brain, liver, spleen, thymus, kidney, adrenal, bladder, heart, ovary, uterus
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring not selected as parental animals were sacrificed at 4 days of age.
- These animals were subjected to postmortem examinations (macroscopic examination) as follows: external malformations, visceral malformations
Statistics:
Yates test, Mann-Whitney U test, Fisher exact test, Bartlett test, Dunnett test, Scheffe test, Kruskal-Wallis test
Reproductive indices:
- Copulation index = (number of copulated pairs / number of pairs mated) x 100
- Fertility index = (number of pregnant animals / number of copulated pairs) x 100
- Gestation index = (number of pregnant females with pups alive / number of pregnant females) x 100
- Implantation index = (number of implantation sites / number of corpora lutea) x 100
Offspring viability indices:
- Delivery index = (number of pups born / number of implantation sites) x 100
- Birth index = (number of pups alive on day 0 / number of implantation sites) x 100
- Live birth index = (number of pups alive on day 0 / number of pups born) x 100
- Sex ratio on day 0 = (number of males pups alive on day 0 / number of female pups alive on day 0) x 100
- Viability index = (number of pups alive on day 4 / number of pups alive on day 0) x 100
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
ignificant increase was observed in 100 mg/kg bw/day group, not considered to be related to treatment
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
significant decrease was observed in 100 mg/kg bw/day group during lactation, not considered to be related to treatment
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
significant decrease of mean corpuscular haemoglobin concentration in the 300 and 1000 mg/kg bw/day groups; fndings are not dose-dependent
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
significant decreased ALP level in all dose groups; significant decreased glucose level in the 1000 mg/kg bw/day group; signifant increase in chloride, decrease in calcium content and in total protein in the 300 mg/kg bw/day group; fndings are not dose-dependent and considered incidental
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
observations were considered incidential
Histopathological findings: neoplastic:
no effects observed
Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
- Males: no deaths or abnormalities in general condition were observed in any of the treated groups
- Females: no deaths were observed in any treated group

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Males: significant increase (p<0.01) compared to control was observed in 100 mg/kg group between 8 - 15 days and between 15 - 29 days. However, since no change was observed in other groups, the effect was not considered to be related to the dosing of compound. No changes in food consumption related to the dosing of compound were observed.
- Females: no significant changes in body weight were noted and there were no changes related to the dosing of compound in body weight gain and food consumption. A significant decrease (p<0.01) compared to control was observed during lactation in 100 mg/kg dosing group. However, since no other changes in food consumption were noted in 300 mg/kg and 1000 mg/kg dosing groups, the effect was not considered to be related to the dosing of compound.

HAEMATOLOGY
- Males: a significant decrease (p<0.01) of mean corpuscular haemoglobin concentration (MCHC) compared to control was found in the 300 and 1000 mg/kg bw/day dose group, respectively (control: 34.7 ± 0.5%; 100 mg/kg bw/day: 34.4 ± 0.2%; 300 mg/kg bw/day: 33.9 ± 0.4%**; 1000 mg/kg bw/day: 33.9 ± 0.3%**; **p<0.01). No other differences were noted.

CLINICAL CHEMISTRY
- Males: a significant decreased ALP level (p<0.05) compared to control was found in all dosing groups (control: 237 ± 45 U/L; 100 mg/kg bw/day: 200 ± 35 U/L*; 300 mg/kg bw/day: 195 ± 35 U/L*; 1000 mg/kg bw/day: 197 ± 41 U/L*; *p<0.05). A significant decreased glucose level (p<0.05) compared to control was found in the high dose group (control: 152 ± 16 mg/dL; 1000 mg/kg bw/day: 135 ± 14 mg/dL*; *p<0.05). While a significant increase in chloride (p<0.05) was found in the 300 mg/kg bw/day group, a significant decrease in calcium content (p<0.01) and in total protein (p<0.05) was noted in this group. However, all the findings lack a dose-dependency and were regarded as incidental.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
There were no significant differences in number of corpora lutea, implantation rate, number of implantations and all other reproductive parameters in all treated groups compared to the control group.

ORGAN WEIGHTS (PARENTAL ANIMALS)
Males: in 100 mg/kg bw males, the actual weight ratio of liver weight (p <0.05) compared to control values were increased. No other significant differences were found in all groups. Thus, this finding lacks a dose-dependency and was regarded as incidental.
- Females: kidney weight was significantly reduced (p<0.05) in the 300 mg/kg bw group. No other weight changes were noted. Thus, this finding lacks a dose-dependency and was regarded as incidental.

GROSS PATHOLOGY (PARENTAL ANIMALS)
Males:
- Heart: myocardial degeneration was noted in one animal of the control group and the 1000 mg/kg bw group, respectively
- Liver: periportal fatty change of the liver was found in all animals of the control and the high dose group. Focal necrosis was noted in one animal of the high-dose group.
- Spleen: all animals of the control and the high-dose group showed brown pigment deposits of the spleen as well as extramedullary haematopeiosis
- Kidney: while fibrosis was only found in 1/13 animals of the control group, eosinophilic bodies and basophilic tubuli in the cortex were noted in animals of both control and high dose groups, respectively
- Adrenal gland: one animal of the high-dose group showed fibrosis
- Testis: atrophy of the seminiferous tubules was observed in two animals of the 1000 mg/kg dose group, with one animal affected on both sides and one side affected in the other animal. No other abnormalities were noted.
- Epididymis: abnormal sperm granuloma was found in one animal of the control group
- Brain: no abnormal findings noted
- Thymus: no abnormal findings noted
- Bladder: no abnormal findings noted

Females:
- Brain: one animal of the high-dose group showed abnormal mineral deposition in the thalamus
- Liver: focal necrosis and fibrosis was found in one animal of the control group, while one animal of the high-dose group showed only focal necrosis
- Spleen: brown pigmentation and extramedullary haematopeiosis was found in animals of the control and the high-dose group, but no varying degrees of frequency was observed
- Thymus: atrophy and bleeding was observed in animals of both groups, control and high-dose group, respectively
- Kidney: basophilic tubuli of the cotex and a dilatation of the renal pelvis was seen in the control and high-dose group
- Adrenal gland: cortical necrosis was observed in one animal of the control group
- Heart: no abnormal findings noted
- Bladder: no abnormal findings noted
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no adverse substance -related systemic effects and findings on fertility were noted
Critical effects observed:
no
Clinical signs:
not examined
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Food efficiency:
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
VIABILITY (OFFSPRING)
No change in number of birth, birth rate, infant live birth rate, infant survival and birth rates at day 4 was found in all dose groups compared to the values of the control group.

BODY WEIGHT (OFFSPRING)
No difference in bodyweight of pups of the dose groups and the control group was found.

GROSS PATHOLOGY (OFFSPRING)
No morphological abnormalities were found in all groups.
Dose descriptor:
NOAEL
Remarks:
developmental
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 substance-related findings were noted
Critical effects observed:
no
Reproductive effects observed:
no

Table 1. Summary of development up of pups from dams treated orally with docosanoic acid in the combined repeat dose and reproductive/developmental toxicity screening test: Mean ± SD (N)

Dose group (mg/kg bw/day)

0

100

300

1000

Number of pregnant females

13

12

12

13

Number of pregnant females with live pups

13

12

12

13

Gestation index

100

100

100

100

Gestation length in days

22.2±0.4 (13)

22.4±0.5 (12)

22.3±0.5 (12)

22.2±0.4 (13)

Number of corpora lutea

16.6±1.6 (13)

16.7±1.8 (12)

16.8±2.3 (12)

16.2±1.5 (13)

Number of implantation sites

16.1±1.5 (13)

15.8±2.1 (12)

16.0±1.5 (12)

15.2±3.1 (13)

Implantation index

96.9±5.1 (13)

94.8±7.1 (12)

96.1±6.0 (12)

93.1±15.2 (13)

Day 0 of lactation

 

 

 

 

Number of pups born

15.2±1.6 (13)

14.8±2.2 (12)

15.2±1.5 (12)

14.3±2.7 (13)

Delivery index

94.5±8.0 (13)

93.5±3.9 (12)

94.9±4.3 (12)

94.9±6.2 (13)

Number of live pups

14.9±1.6 (13)

14.3±2.2 (12)

15.1±1.6 (12)

14.1±2.7 (13)

Birth index

93.1±8.8 (13)

90.7±8.7 (12)

94.3±5.1 (12)

93.5±7.2 (13)

Live birth index

98.5±2.8 (13)

97.0±8.5 (12)

99.4±2.2 (12)

98.5±4.0 (13)

Sex ration on day 0

50.0±11.3 (13)

46.7±9.8 (12)

54.8±12.3 (12)

48.9±13.4(13)

Day 4 of lactation

 

 

 

 

Number of live pups

14.7±1.4 (13)

13.0±4.7 (12)

15.1±1.6 (12)

14.1±2.7 (13)

Viability index

98.6±2.7 (13)

89.4±28.8 (12)

100.0±0.0 (12)

100.0±0.0 (13)

Sex ration on day 4

49.8±11.5 (13)

46.3±10.3 (11)

54.8±12.3 (12)

48.9±13.4(13)

Endpoint:
screening for reproductive / developmental toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
5 Sep 1989 - 26 Okt 1989
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
yes
Remarks:
male rats were not exposed to the test substance and not included in any examinations, female rats were only exposed 7 days prior to mating, no data on test substance purity, parental organ weights were not determined
GLP compliance:
yes
Remarks:
FDA "Good Laboratory Practice Regulations"
Limit test:
no
Species:
rat
Strain:
other: Crl:CD® (SD) BR
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories, Portage, Michigan, USA
- Age at study initiation: males / females: about 12 weeks
- Weight at study initiation: males: no data; females: 248.3 - 255.4 g
- Fasting period before study: no data
- Housing: individually in wire-bottomed stainless-steel cages suspended above absorbent paper liners, except during the cohabition and post-parturition period
- Diet: certified rodent chow 5002 (Ralston Purina), ad libitum
- Water: local water that had been passed through a reverse osmosis membrane supplemented with chlorine (max. of 0.6 ppm) as bacteriostat, ad libitum
- Acclimation period: 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 - 26
- Humidity (%): 35 - 65
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: 19 September 1989 To: 26 October 1989
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
- weekly (prepared solutions were refrigerated and protected form light in amber glass bottles)

VEHICLE
- Concentration in vehicle: 40, 200 and 400 mg/mL
- Amount of vehicle (if gavage): 5 mL/kg bw, adjusted daily for body weight changes
- Lot: MAR 16 90B and AUG 11 90B
Details on mating procedure:
- M/F ratio per cage: 1/1
- Length of cohabitation: maximum of seven days
- Proof of pregnancy: vaginal plug and/or sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged (how): individually
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Two samples (5 mL each) of each concentration were reserved on the first day the dosage solutions were prepared and also from the last batches prepared. One of every two samples reserved were shipped to the sponsor for analysis. The results of these analyses remained at the sponsor. No further information was given.
Duration of treatment / exposure:
females: about 40 days (7 days prior to mating until day 4 of lactation)
males: untreated
Frequency of treatment:
daily, 7 days/week
Dose / conc.:
200 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
Dose / conc.:
2 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: no data (Dosages tested were selected by the sponsor. No further details were given.)
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least twice daily for viability, proof of pregnancy and for duration of gestation/parturition; during acclimation period, rats were observed three times for clinical signs and general health and additionally for pharmacotoxic effects daily during the dosage period (immediately prior to and then within 60 minutes after intubation)

BODY WEIGHT: Yes
- Time schedule for examinations: once weekly during acclimation period and daily thereafter

FOOD CONSUMPTION: Yes
- Time schedule for examinations: weekly prior to mating; on days 0, 6, 10, 14, 16, 21 and 25 of presumed gestation; days 1 and 4 of lactation
Oestrous cyclicity (parental animals):
No data
Sperm parameters (parental animals):
No data
Male rats were used only as breeders, and were not given the test substance. They were not considered to be a part of the test system and therefore not evaluated.
Litter observations:
PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
litter numbers, viability, clinical observations, body weights, sex ratios

GROSS EXAMINATION OF DEAD PUPS:
yes, gross external physical anomalies
Postmortem examinations (parental animals):
GROSS NECROPSY
- all dams were examined for gross lesions of the thoracic and abdominal viscera (dams that did not deliver litters were sacrificed on day 25 of presumed gestation)
- male animals were not examined

HISTOPATHOLOGY / ORGAN WEIGHTS
Tissues with gross lesions present were preserved in neutral buffered 10% formalin.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring was sacrificed on day 4 of lactation.
- These animals were subjected to postmortem examinations as follows: all animals

GROSS NECROPSY
- Gross necropsy consisted of examination for gross lesions of the thoracic and abdominal viscera. Lungs were removed and immersed in water. Pups with lungs sank were considered stillborn; pups with lungs that floated were considered liveborn and to have died shortly after birth.

HISTOPATHOLOGY / ORGAN WEIGHTS
Ovaries and tissues with gross lesions present were preserved in neutral buffered 10% formalin.
Statistics:
Maternal and pup incidence data were analyzed using the variance test for homogeneity of the binomial distribution.
Baseline maternal body weight data, body weight changes during the premating, gestation and lactation periods, feed consumption data and litter averages for pup body weights and percentage of male pups were analysed using Bartlett´s test of homogeneity of variances and the analysis of variance, when appropriate (i.e. Bartlett´s Test was not significant). If the analysis of variance was significant, Dunnett´s test was used to identify the statistical significance of individual groups. If the analysis of variance was not appropriate (i.e. Bartlett´s Test was significant), the Kruskal-Wallis test was used when 75% or fewer ties were present; when more than 75% ties were present, the Fisher´s Exact test was used. In cases where the Kruskal -Wallis test was statistically significant, Dunn´s method of multiple Comparisons was used to identify the statistical significance of individual groups.
Natural delivery parameters involving discrete data were evaluated using the Kruskal-Wallis test procedures previously described.
Reproductive indices:
- Fertility index = (number of pregnant female rats/ rats mated)
- Gestation index = (number of females with live litters/ pregnant rats)
- Implantation sites
Offspring viability indices:
- Viability index = (pups surviving 4 days / liveborn pups)
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
see "Details on results (P0)"
Mortality:
mortality observed, treatment-related
Description (incidence):
1000 mg/kg bw/day: 1 female died (non-adverse) ; 2000 mg/kg bw/day: 3 females died
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
see "Details on results (P0)"
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
see "Details on results (parental animals)"
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
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
200 mg/kg bw/day: Rales occured during premating and gestation.
1000 mg/kg bw/day: One female died following an intubation accident. Rales occured during premating, gestation and lactation. Excess salivation was observed during premating, gestation and lactation. Other clincial observations included urine-stained abdominal fur, chromorrhinorrhea, oral exudate, ungroomed coat, abscence of motor activity and ataxia. These effects were considered to be treatment-related.
2000 mg/kg bw/day: Rales and excess salivation occured during premating, gestation and lactation. 2 animals died during the premating dose and one animal on day 3 of gestation. In addition to rales and excess salivation, these animals had urine-stained abdominal fur, ungroomed coat, chromorrhinorrhea and/or decreased motor activity as well as ataxia. These effects were considered to be treatment-related.
In addition, dental problems, red vaginal exudate, a skin lesion and localized alopecia were observed but not related to any treatment group.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
1000 mg/kg bw/day: On day 1 to 7 of the study, mean body weight change was lower than in controls (+ 7.8 g vs. + 8.8 g). Relative feed consumption was about 6% lower than in controls. During gestation, maternal body weight changes averaged +140.5 g (vs. 132.8 g in controls) although relative feed consumption was about 6% lower than in controls. During lacatation, maternal body weight changes averaged +12 g (vs. +0.1 g in controls). Relative feed consumption was about 13% increased, when compared to controls.
2000 mg/kg bw/day: Reduced body weight gain, reduced mean absolute body weights (about 2%) and reduced relative feed consumption (- 17% compared to controls) were observed during the whole premating period. During gestation, maternal body weight changes were significantly lower than in controls (+ 126 g vs. 132.8 g). Absolute body weights were about 7% lower when compared to controls. Relative feed consumption tended to be reduced when compared to controls (about 2%). During lactation, maternal body weight changes averaged -14.5 g (vs. +0.1 g in controls). Absolute body weights were about 11% lower when compared to controls. Relative feed consumption was about 5% decreased, when compared to controls.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
No effects on fertility and gestation indices were observed.

GROSS PATHOLOGY (PARENTAL ANIMALS)
200 mg/kg bw/day: One animal had a fluid-filled uterus. This effect was related to estrous cycling and therefore to be non-adverse.
1000 mg/kg bw/day: The animal that was found dead had gross lesions indicative for an intubation accident like bright red, distended lungs and a yellow viscous fluid resembling the vehicle in the trachea.
2000 mg/kg bw/day: One animal found dead prior to scheduled sacrifice had red mottled lungs.
Dose descriptor:
NOAEL
Remarks:
systemic
Effect level:
200 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: mortality; clinical signs; body weights during lactation
Dose descriptor:
NOAEL
Remarks:
fertility
Effect level:
>= 2 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: NOAEL corresponding to the highest dose tested.
Critical effects observed:
not specified
Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
non-adverse
Food efficiency:
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
Behaviour (functional findings):
not examined
Developmental immunotoxicity:
not examined
VIABILITY (OFFSPRING)
No effects were oberved.

CLINICAL SIGNS (OFFSPRING)
No effects were oberved.

BODY WEIGHT (OFFSPRING)
2000 mg/kg bw/day: Litters tended to have reduced body weights (about 7%) on day 4 postpartum.

GROSS PATHOLOGY (OFFSPRING)
No effects were oberved.
Dose descriptor:
NOAEL
Remarks:
developmental
Generation:
F1
Effect level:
>= 2 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: NOAEL corresponding to the highest dose tested.
Critical effects observed:
no
Reproductive effects observed:
no
Endpoint:
extended one-generation reproductive toxicity - basic test design (Cohorts 1A, and 1B without extension)
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The available information comprises adequate, reliable studies (Klimisch score 1 and 2) and consistent studies from reference substances with similar structure and intrinsic properties. Read-across is justified based on a common functional group, common precursors/breakdown products and similarities in toxicological properties (refer to endpoint discussion for further details). The selected study is thus sufficient to fulfil the standard information requirements set out in Annex VIIISection 8.7.1, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006. An extended one-generation reproduction toxicity study as set out in Annex IX-X, Section 8.7.3 does not need to be conducted and is scientifically unjustified, as no adverse effects on reproductive organs or any adverse systemic effects in acute toxicity, repeated dose toxicity, genetic toxicity and developmental toxicity studies were observed and as fatty acids fulfil physiological functions within the body.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Human health effects on toxicity to reproduction are predicted from adequate and reliable data for source substances by read-across to the target substance within the group applying the group concept in accordance with Annex XI, Item 1.5, of Regulation (EC) No 1907/2006.

Fatty acids are found in all living organism fulfilling three fundamental roles. Besides their function as part of molecules like phospholipids and glycolipids important for the cell-structure, they are often precursors of signalling molecules such as prostanoids. The third and best understood role of fatty acid is their role as nutritional energy source. Based on their physiological function within the body no toxicity to reproduction of fatty acids is expected as demonstrated by animal studies with C7 fatty acid (heptanoic acid), C22 fatty acid (docosanoic acid) and fatty acids, tall oil.

 

The toxicity to reproduction of docosanoic acid (CAS# 112-85-6) was evaluated in a combined repeated dose and reproductive/developmental toxicity screening test performed under GLP according to OECD guideline 422 (2002).Groups of 13 male and 13 female Sprague-Dawley rats received daily doses of 100, 300 and 1000 mg/kg bw/day of docosanoic acid by gavage. While the males were treated for 42 days, the females received the test substance from 14 days prior to mating until day 3 of lactation. As result, neither mortality nor abnormalities in general condition were observed. Since no difference in number of corpora lutea, implantation rate, number of implantations and all other reproductive parameters was noted in all treated groups compared to the control group, there were no indications of any adverse effects on reproduction. Thus, the NOAEL toxicity to reproduction was found to be ≥ 1000 mg/kg bw/day.

 A combined reproduction / developmental toxicity screening test performed equivalent to OECD guideline 421 is available for heptanoic acid (CAS# 111-14-8) (1990). 10 female Sprague-Dawley rats were orally treated with 200, 1000 or 2000 mg/kg bw/day of the test substance dissolved in corn oil. Control animals received the vehicle only. Females were treated 7 days prior to mating and afterwards during mating, gestation and lactation until day 4 post-parturition. Male rats were used only as breeders (10 animals/group), and were not given the test substance. They were not considered to be a part of the test system and therefore not evaluated. Examinations on all dams comprised cage side observations, determinations of body weights and food consumption, gross necropsy, histopathology of gross lesions as well as determinations of reproductive indices.

One dam of the 1000 mg/kg bw/day dose group died following an intubation accident. In the 2000 mg/kg bw/day dose group 3 animals died, which was considered to be a treatment-related. Clinical signs included rales in significant numbers of rats of all treated groups over the whole study period. Possibly, this effect is due to gastroesophagal reflux and subsequent aspiration of dosing solution. Furthermore, excess salivation, urine-stained abdominal fur, ungroomed coat, chromorrhinorrhoea and/or decreased motor activity as well as ataxia were observed in the mid and high-dose group animals.

On day 1 to 7 of the study, mean body weight change in animals of the 1000 mg/kg bw/day dose group was lower than in controls (+ 7.8 g vs. + 8.8 g). Concomitantly, relative feed consumption was about 6% lower than in controls. During gestation, maternal body weight changes averaged + 140.5 g (vs. 132.8 g in controls) although relative feed consumption was about 6% lower than in controls. During lactation, maternal body weight changes averaged +12 g (vs. + 0.1 g in controls) and relative feed consumption was about 13% increased, when compared to controls. In the 2000 mg/kg bw/day dose group reduced body weight gain, reduced mean absolute body weights (about 2%) and reduced relative feed consumption (-17% compared to controls) were observed during the whole premating period. During gestation, maternal body weight changes were significantly lower than in controls (+ 126 g vs. + 132.8 g). Absolute body weights were about 7% lower when compared to controls and concomitantly, relative feed consumption tended to be reduced when compared to controls (about 2%). During lactation, maternal body weight changes averaged -14.5 g (vs. +0.1 g in controls). Absolute body weights were about 11% lower when compared to controls and the relative feed consumption was about 5% decreased, when compared to controls.

No effects on fertility and gestation indices as well as average number of implantations were observed. At gross pathology, the only treatment-related effect observed was red mottled lungs in one of the high-dose females.

Based on the results described above, the NOAEL for fertility was determined to be ≥ 2000 mg/kg bw/day. Based on mortality, clinical signs and reduced body weights during lactation in the high-dose group, the NOAEL for maternal toxicity is 200 mg/kg bw/day for females. In contrast no maternal toxic effects were found in two studies similar to OECD 414 performed with heptanoic acid in concentrations up to and including 1000 mg/kg bw/day (1982 and 1983).

In a two-generation reproduction toxicity study similar to OECD guideline 416, Sprague-Dawley rats (30 females per group and 15 males per group) were administered 5% and 10% fatty acids, tall oil (CAS# 61790-12-3) in the diet (corresponding to 2500 and 5000 mg/kg bw/day) as cited in a secondary source (Pine Chemical Association, 2004). The parental animals were fed the test substance, which consists predominantly of C18 unsaturated and saturated fatty acids, starting at 20 days pre-mating. Treatment of the parental generation was through the weaning of the first generation (F1). After weaning, 20 F1 males and 20 F1 females per group were maintained on the parental diet and mated at 100 days of age. The delivered F2 pups were followed until weaning. Parameters evaluated included F1 reproductive parameters, F1 fertility, viability, lactation and gestation indices. Haematology, serum chemistry, urinalysis, and organ weights for F1 animals, gross pathology of F1 and F2 animals and microscopic pathology of various organs of the F2 pups were performed.

There were no treatment-related effects on reproductive performance, the number of liveborn or still-born F1 litters and pups, or weaning weight of the F1 pups. No treatment-related changes in fertility, viability, lactation, or gestation indices were noted. No changes in haematology, clinical chemistry and urinalysis parameters and organ weights were found. Gross and microscopic pathology revealed no treatment-related effects. Thus, fatty acids, tall oil had no effect on the reproductive or developmental capabilities of rats at doses up to and including 5000 mg/kg bw/day.

 

No adverse effects on reproductive organs were observed in 28-day or 90-day repeated dose toxicity studies performed with members of the fatty acids category.

In a subchronic study rats received oleic acid (CAS# 112-80-1) at dose levels up to 12500 mg/kg bw/day (1969). Gross pathology and histopathology included the determination of gonads weight as well as examination of testes, seminal vesicles, ovaries and uterus. No changes in absolute or relative gonads weight were noted and no gross and histopathological findings were observed in any of the examined reproductive tissues and organs. Moreover, in a subacute toxicity study (1983) with fatty acids, C8-18 and C18-unsatd, distn. residues (CAS# 70321-72-1) no adverse effects on reproductive organs up to and including the limit dose of 1000 mg/kg bw/day was noted, which was demonstrated by a lack of changes in gonads weight and the absence of histopathological findings in epididymides, seminal vesicles, testes, ovaries and uterus.

Since no effects on fertility were noted in the reproduction / developmental toxicity screening studies with heptanoic acid and docosanoic acid and since no adverse effects on reproductive organs were observed in 28- and 90-day repeated dose toxicity studies, an extended one-generation reproductive toxicity study is therefore scientifically unjustified taking also into account the low toxicological activity (no evidence of toxicity seen in any of the relevant routes of exposure) and the physiological functions of fatty acids within the body.

Since all the members of the category show similar structural and toxicological properties the results of individual members are also valid for other fatty acids within the category. Moreover, a substance-specific adjustment of the NOAEL for the individual category members is not required. An overall NOAEL for fertility for all fatty acids within the category of 1000 mg/kg bw/day is chosen as “worst case” among the available studies.

Effects on developmental toxicity

Description of key information

Key studies on developmental toxicity/teratogenicity by the oral route are available for the following category members:
Prenatal Developmental Toxicity Study (OECD 414), rat: NOAEL oral (develop. toxicity/teratogenicity) ≥1000 mg/kg bw/day; CAS# 111-14-8, C7
Prenatal Developmental Toxicity Study (OECD 414), rat: NOAEL oral (develop. toxicity/teratogenicity) ≥1500 mg/kg bw/day; CAS# 112-05-0, C9
Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test (OECD 422), rat: NOAEL oral (developmental toxicity/teratogenicity) ≥1000 mg/kg bw/day; CAS# 112-85-6, C22

No hazard for developmental toxicity/teratogenicity was identified for fatty acids.

No data are available for developmental toxicity/teratogenicity after dermal exposure and inhalation, respectively.

Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11 Mai - 24 Jun 1982
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
test substance was administered solely in the period of GD 6-15
GLP compliance:
not specified
Limit test:
yes
Species:
rat
Strain:
other: Crl:COBS, CD® (SD) BR
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories, Inc., Kingston, New York, USA
- Age at study initiation: approx. 14 weeks (mating age)
- Housing: individually in elevated wire-mesh cages
- Diet: Purina Rodent Laboratory Chow ®, ad libitum
- Water: tap water, ad libitum
- Acclimation period: 9 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): approx. 24
- Humidity (%): 57 ± 4.8
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: 11 Mai 1982 To: 24 Jun 1982
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Dosing solutions were prepared weekly by mixing appropriate amounts of the test item with corn oil on a magnetic stirrer (g/v) and transferred to amber bottles. The prepared dilutions were well shaken before use and animals were dosed while solutions were mixed on a magnetic stirrer.

Dosing volumes were based on individual body weights from the most weighing interval. Day 15 dosing was based on the body weight of Day 12.


Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
- Impregnation procedure: cohoused
- M/F ratio per cage: 1/2
- Length of cohabitation: up to 3 weeks
- After 10 days, females were rotated
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
Duration of treatment / exposure:
Day 6 - 15 of gestation
Frequency of treatment:
daily, 7 days/week
Duration of test:
20 days (GD 0-20)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
22 females
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The applied dose showed no maternal toxicity in a previous dose-range finding study (1982).
Maternal examinations:
CLINICAL OBSERVATIONS: Yes
- Time schedule: daily
- Parameters checked: e.g. alopecia, bloody crust, wheezing, labored respiration, urine stains, rough haircoat, stains on fur, salivation, corneal defect, wasting feed, water spillage, hunched, dyspnea, red discharge from vagina, soft feces

BODY WEIGHT: Yes
- Time schedule for examinations: days 0, 6, 9, 12, 15 and 20 of gestation

FOOD CONSUMPTION : Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food: Yes
- Time schedule for examinations: Days 6 - 8, 9 - 11, 12 - 14, 15 - 17 and 18 - 20 of gestation

WATER CONSUMPTION: Yes
- Time schedule for examinations: Days 6 - 8, 9 - 11, 12 - 14, 15 - 17 and 18 - 20 of gestation

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: uterus, oavary, cecum, lung, kidney, stomach, liver, pancreas, adrenals, brain, pituitary, thymus, spleen, eyes, lymph nodes (mesenteric, cervical, mandibular)

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 resorptions: Yes (early and late resorptions)
- Other: Nongravid uterine weights (with ovaries attached): Yes
Fetal examinations:
- External examinations: Yes: [all per litter]
- Skeletal examinations: Yes: [two-third per litter]
- Other: Visceral examinations: Yes [one-third per litter]; body weight and crown-rump distance [all per litter]
Statistics:
Mean values and standard deviations were calculated from the examined parameters.
Survival was analysed by the National Cancer Institute Package (Tomas, Breslow and Gart, 1977). The mean maternal body weight changes (days 0 - 6, 6 - 15, 15 - 20 and 0 - 20), mean maternal food and water consumptions), percent males per litter, mean fetal body weights and lengths, fetal viability, percent resorptions, implantation efficiency, gravid and non-gravid uterine weights and the incidence of visceral and skeletal anomalies and variants were analysed by Box´s test for homogeneity of variances. This test was followed by one-way classification analysis of variance (ANOVA) if the variances proved to be homogeneous. If the variances proved to be heterogeneous, a rank transformation of data was performed, which was followed by Box´s test and ANOVA. If ANOVA of untransformed or transformed data was significant, a Dunnett´s T-Test was used for control vs treatment group mean comnparisons. Pregnancy rates were analysed by a test of multiple proproportions using one degree of freedom Chi-square test with Yate´s continuity correction (Fleiss 1981). All pairwise comparisons were evaluated at the 0.5% probability (one-tailed) level.
Indices:
Mean incidence of resorptions (%): group mean of [(resorptions per litter/implantations per litter) x 100]
Mean incidence of fetal mortality (%): group mean of [(dead fetuses per litter/implantations per litter) x 100]
Mean incidence of fetal viability (%): group mean of [(live fetuses per litter/implantations per litter) x 100]
Mean incidence of visceral anomalies(%): group mean of [(number fo fetuses with anomalies per litter/number of fetuses examined viscerally per litter) x 100]
Mean incidence of visceral variants (%): group mean of [(number fo fetuses with variants per litter/number of fetuses examined viscerally per litter) x 100]
Mean incidence of skeletal anomalies (%): group mean of [(number fo fetuses with anomalies per litter/number of fetuses examined skeletally per litter) x 100]
Mean incidence of skeletal variants (%): group mean of [(number fo fetuses with variants per litter/number of fetuses examined skeletally per litter) x 100]
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
effects were considered incidental
Mortality:
mortality observed, non-treatment-related
Description (incidence):
one female died, not considered treatment-related
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):
no effects observed
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):
effects were considered incidental
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
not specified
Early or late resorptions:
not specified
Dead fetuses:
no effects observed
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:
no effects observed
Details on maternal toxic effects:
MORTALITY AND CLINICAL OBSERVATIONS
One animal in the treatment group was found dead on day 8 of gestation. However, the survival rate of treated animals (95.5%) were comparable to control (100%).
No increased incidence of clinical observations was noted. Wheezing was observed in three treated rats during GD 6-15 and in two treated rats after treatment (GD 16-20). One treated rat had a rough haircout. In the control group, alopecia (neck), corneal defect, wasting feed and water spillage was each observed in one animal.


BODY WEIGHT CHANGE, FOOD AND WATER CONSUMPTION
No effects on body weights, food and water consumption were noted. The mean body weight change (Day 0-20 of gestation) was 104.2 ± 12.5 g for the treated females compared to 106.9 ± 14.7 g for the control group. The treated rats had a total food consumption intake (Day 9 -20 of gestation) of 243.0 ± 31.3 g (controls: 249.9 ± 43.7).

GROSS PATHOLOGY
The effects noted in gross pathology examinations are present in control and test animals in low frequencies and are therefore considered to be incidential in nature and showed no relation to treatment. In detail, control and test animals revealed anomalies of the kidneys (pelvises dilated, firm nodules on surface, depressed area in cortex) and red focal areas in the lung. Further, one test animal each showed yellow areas on the liver surface, reddened/enlarged cervical lymph nodes and a thin and pale nonglandular mucosa and smooth glandular mucosa. One control animal had clear raised areas on the surface of eyes.

OTHER
The pregnancy rate was 100% in the 1000 mg/kg bw/day dose group and in control animals.
No alterations in the number of corpora lutea, implantations and mean implantation efficiencies (number of implantations per number of corpura lutea) and resorptions were observed .
Gravid and nongravid uterine weights were comparable between the control and dose group (table 1).
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
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): not examined
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
not examined
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Details on embryotoxic / teratogenic effects:
No alterations in the number of live and dead fetuses were observed. Fetal viability, size and sex remained unchanged.
Anomalies determined in gross pathology, visceral and skeleton examinations were present in both, control and test animals and are therefore not considered as treatment-related effects (for details, see table 2).

Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: teratogenicity and development
Abnormalities:
no effects observed
Developmental effects observed:
no

Table 1: Ovarian, Uterine and Litter Data

Parameter

Control

1000 mg/kg bw/day

Mean uterine weights

Gravid

69.48 ± 13.953

67.63 ± 11.91

Mean uterine weights

Nongravid

6.52 ± 1.424a

7.04 ± 1.982a

Corpora lutea (mean number)

15.9

14.7

Implantations (number)

13.5

12.8

Mean implantation efficiency (indeces)

85.7a

88a

Resorptions (number)

1.2

0.6

Mean incidence of resorptions (%)

9.2a

4.3a

Dead fetuses (number)

0.0

0.0

Mean incidence of fetal mortality (%)

0.0a

0.0a

Live fetuses (number)

12.4 ± 2.56

12.2 ±1.91

Mean incidence of fetal viability (%)

90.8a

95.7a

Incidences were calculated on a per litter basis.

Data are shown as means.

a: Data analysed following rank transformation

 

Table 2: Examination of the fetuses

Parameter

Control

1000 mg/kg bw/day

Live fetuses

Males

Mean body weight (g)

3.58 ± 0.201b

3.46 ± 0.264b

Mean crown-rump distance (cm)

3.8 ± 0.15b

3.8 ± 0.19b

Females

 

 

Mean body weight (g)

3.39 ± 0.186b

3.32 ± 0.248b

Mean crown-rump distance (cm)

3.7 ± 0.14b

3.7 ± 0.18b

Males per litter (Mean %)

54.0 ± 18.31

47.4 ± 13.62

Gross pathology findings:

Number of fetuses examined

191

175

Number of fetuses appearing normal

169

166

Skull

0

0

Palate

0

0

Cleft palate

0

0

Heart

0

0

Kidney

0

0

Ureter

0

0

Dilated

16

7

Hydroureter

0

1

Undulated

2

1

Umbilical hernia

1

0

Situs inversus

1

0

Hindleg

0

0

Small

1

0

Exencephaly

0

0

Tail missing

0

0

Mean incidence values for visceral findings

Number of litters

22

22

Number with anomalies

0a

0.05 ± 0.213a

Incidence of anomalies (%)c

0a

1 ± 5.3a

Number with variants

0.1 ± 0.29a

0.1 ± 0.29a

Incidence of variants (%)d

3 ± 9.7a

3 ± 11.7a

Mean incidence values for skeletal findings

Number of litters

22

22

Number with anomalies

0a

0a

Incidence of anomalies (%)e

0a

0a

Number with variants

3 ± 2.3a

3 ± 2.2a

Incidence of variants (%)f

39 ± 25.2a

31 ± 24.6a

Visceral Findings

Number of fetuses examined/number of litters

81/22

80/22

Number of fetuses with anomalies/number of litters with anomalies

0/0

1/1

Number of anomalies

0

1

Hydroureter

0

1

Number of fetuses with variants/number of litters with anomalies

2/2

2/2

Number of variants

2

3

Dilated renal pelves

2

1

Dilated ureters

0

2

Fetus small

0

0

Small tongue

0

0

Skeletal Findings

 

 

Number of fetuses examined/number of litters

191/22

175/22

Number of fetuses with anomalies/number oflitters with anomalies

0/0

0/0

Number of anomalies

0

0

Number of fetuses with variants/number of litters with variants

73/19

57/18

Number of variants

110

89

Skull:

 

 

Intraparietal-incomplete ossification/nonossified

21

20

Supraoccipital-incomplete ossification/nonossified

14

11

Hyoid

25

22

Supraoccipital - nonfused

0

2

Supraoccipital - lopsided

0

0

Supraoccipital - small

0

0

Small

0

0

Nasal bones appear short

0

0

Rib cage:

 

 

Parletals – irregular edges

2

4

Ribs – angulated

2

1

Ribs – 13thpair small

1

0

Ribs – first pair small

0

0

Ribs – forked

0

0

Vertebrae:

 

 

Centra – incomplete ossification / nonossified

32

19

Centra – nonfused

10

5

Centra small

0

0

Centra – missing

0

0

Sternabrae – bipartite

1

0

Hemivertebrae

0

0

Vertebrae – less than 3 ossified

0

0

No of vertebrae below 3rdsacral

0

0

Sacral vertebrae – nonossified

0

0

Halaligned centra ans arches / caudals

0

0

Caudals – less than 3 ossified

0

1

Caudals – incomplete ossification

0

0

Arches – nonossified

0

0

Pelvis:

 

 

Pelvic girdle – incomplete ossification

1

2

Pelvic girdle – small

0

0

Pubis – incomplete ossification/nonossified

1

1

Pubis – small

0

0

Ischium – incomplete ossification / nonossified

0

0

Data are shown as mean ± standard deviations.

a: Incidences were calculated on a per litter basis.

b: Data analysed following rank transformation

c: Mean Incidence of Visceral Anomalies (%) – Group mean of ((number of fetuses with anomalies per litter/number of fetuses examined viscerally per litter)x100)

d: Mean Incidence of Visceral Variance (%) – Group mean of ((number of fetuses with variants per litter/number of fetuses examined viscerally per litter)x100)

e: Mean Incidence of Skeletal Anomalies (%) – Group mean of ((number of fetuses with anomalies per litter/number of fetuses examined skeletally per litter)x100)

f: Mean Incidence of Skeletall Variance (%) – Group mean of ((number of fetuses with variants per litter/number of fetuses examined skeletally per litter)x100)

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11 Mai - 24 Jun 1982
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
test substance was administered solely in the period of GD 6-15
GLP compliance:
not specified
Limit test:
yes
Species:
rat
Strain:
other: Crl:COBS, CD® (SD) BR
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories, Inc., Kingston, New York, USA
- Age at study initiation: approx. 14 weeks (mating age)
- Housing: individually in elevated wire-mesh cages
- Diet: Purina Rodent Laboratory Chow ®, ad libitum
- Water: tap water, ad libitum
- Acclimation period: 9 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): approx. 24
- Humidity (%): 57 ± 4.8
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: 11 Mai 1982 To: 24 Jun 1982
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Dosing solutions were prepared weekly by mixing appropriate amounts of the test item with corn oil on a magnetic stirrer (g/v) and transferred to amber bottles. The prepared dilutions were well shaken before use and animals were dosed while solutions were mixed on a magnetic stirrer.

Dosing volumes were based on individual body weights from the most weighing interval. Day 15 dosing was based on the body weight of Day 12.


Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
- Impregnation procedure: cohoused
- M/F ratio per cage: 1/2
- Length of cohabitation: up to 3 weeks
- After 10 days, females were rotated
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
Duration of treatment / exposure:
Day 6 - 15 of gestation
Frequency of treatment:
daily, 7 days/week
Duration of test:
20 days (GD 0-20)
Dose / conc.:
1 500 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
22 females
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The applied dose showed no maternal toxicity in a previous dose-range finding study (1982).
Maternal examinations:
CLINICAL OBSERVATIONS: Yes
- Time schedule: daily
- Parameters checked: e.g. alopecia, bloody crust, wheezing, labored respiration, urine stains, rough haircoat, stains on fur, salivation, corneal defect, wasting feed, water spillage, hunched, dyspnea, red discharge from vagina, soft feces

BODY WEIGHT: Yes
- Time schedule for examinations: days 0, 6, 9, 12, 15 and 20 of gestation

FOOD CONSUMPTION : Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food: Yes
- Time schedule for examinations: Days 6 - 8, 9 - 11, 12 - 14, 15 - 17 and 18 - 20 of gestation

WATER CONSUMPTION: Yes
- Time schedule for examinations: Days 6 - 8, 9 - 11, 12 - 14, 15 - 17 and 18 - 20 of gestation

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: uterus, oavary, cecum, lung, kidney, stomach, liver, pancreas, adrenals, brain, pituitary, thymus, spleen, eyes, lymph nodes (mesenteric, cervical, mandibular)

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 resorptions: Yes (early and late resorptions)
- Other: Nongravid uterine weights (with ovaries attached): Yes
Fetal examinations:
- External examinations: Yes: [all per litter]
- Skeletal examinations: Yes: [two-third per litter]
- Other: Visceral examinations: Yes [one-third per litter]; body weight and crown-rump distance [all per litter]
Statistics:
Mean values and standard deviations were calculated from the examined parameters.
Survival was analysed by the National Cancer Institute Package (Tomas, Breslow and Gart, 1977). The mean maternal body weight changes (days 0 - 6, 6 - 15, 15 - 20 and 0 - 20), mean maternal food and water consumptions), percent males per litter, mean fetal body weights and lengths, fetal viability, percent resorptions, implantation efficiency, gravid and non-gravid uterine weights and the incidence of visceral and skeletal anomalies and variants were analysed by Box´s test for homogeneity of variances. This test was followed by one-way classification analysis of variance (ANOVA) if the variances proved to be homogeneous. If the variances proved to be heterogeneous, a rank transformation of data was performed, which was followed by Box´s test and ANOVA. If ANOVA of untransformed or transformed data was significant, a Dunnett´s T-Test was used for control vs treatment group mean comnparisons. Pregnancy rates were analysed by a test of multiple proproportions using one degree of freedom Chi-square test with Yate´s continuity correction (Fleiss 1981). All pairwise comparisons were evaluated at the 0.5% probability (one-tailed) level.
Indices:
Mean incidence of resorptions (%): group mean of [(resorptions per litter/implantations per litter) x 100]
Mean incidence of fetal mortality (%): group mean of [(dead fetuses per litter/implantations per litter) x 100]
Mean incidence of fetal viability (%): group mean of [(live fetuses per litter/implantations per litter) x 100]
Mean incidence of visceral anomalies(%): group mean of [(number fo fetuses with anomalies per litter/number of fetuses examined viscerally per litter) x 100]
Mean incidence of visceral variants (%): group mean of [(number fo fetuses with variants per litter/number of fetuses examined viscerally per litter) x 100]
Mean incidence of skeletal anomalies (%): group mean of [(number fo fetuses with anomalies per litter/number of fetuses examined skeletally per litter) x 100]
Mean incidence of skeletal variants (%): group mean of [(number fo fetuses with variants per litter/number of fetuses examined skeletally per litter) x 100]
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
effects were considered incidental
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):
no effects observed
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):
effects were considered incidental
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
not specified
Early or late resorptions:
not specified
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
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:
no effects observed
Details on maternal toxic effects:
MORTALITY AND CLINICAL OBSERVATIONS
All animals survived to the end of the study.
No increased incidence of clinical observations was noted. Wheezing, water spillage, alopecia, sores (front leg) and blody discharge from vagina was each noted in one treated rat during GD 6-15. In the control group, alopecia (neck), corneal defect, wasting feed and water spillage was each observed in one animal.


BODY WEIGHT CHANGE, FOOD AND WATER CONSUMPTION
No effects on body weights, food and water consumption were noted. The mean body weight change (Day 0-20 of gestation) was 100.4 ± 32.5 g for the treated females compared to 106.9 ± 14.7 g for the control group. The treated rats had a total food consumption intake (Day 9 -20 of gestation) of 243.0 ± 20.4 g (controls: 249.9 ± 43.7).

GROSS PATHOLOGY
The effects noted in gross pathology examinations are present in control and test animals in low frequencies and are therefore considered to be incidential in nature and showed no relation to treatment. In detail, control and test animals revealed anomalies of the kidneys (pelvises dilated, firm nodules on surface, rough surface, pale) and red focal areas in the lung. Further, one test animal each showed dark pinpoint foce in the liver, an uterus distended with clear fluid and enlarged mandibular lymph nodes. One control animal had clear raised areas on the surface of eyes.

OTHER
The pregnancy rate was 100% in the 1500 mg/kg bw/day dose group and in control animals.
No alterations in the number of corpora lutea, implantations and mean implantation eficiencies (number of implantations per number of corpura lutea) and resorptions were observed .
Gravid and nongravid uterine weights were comparable between the control and dose group (table 1).
Dose descriptor:
NOAEL
Effect level:
>= 1 500 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
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): not examined
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
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:
effects observed, non-treatment-related
Description (incidence and severity):
6 fetuses from one dam revealed cleft palate at gross pathology (4 pups) and at visceral examinations (2 pups), considered not treatment-related
Details on embryotoxic / teratogenic effects:
No alterations in the number of live and dead fetuses were observed. Fetal viability, size and sex remained unchanged.
Gross pathology findings in animals receiving skeletal examinations included incidence of cleft palate in 4 pups of the same dam. This finding is rather by chance, since only one litter is affected.
A higher mean incidence of visceral anomalies was noted due to a cleft palate observed in two fetuses from the same litter (for details, see table 2). However, this increase was not statistically significant. These two fetuses belong to the same litter as those fetuses, which revealed cleft palate at gross pathology.

Dose descriptor:
NOAEL
Effect level:
>= 1 500 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: teratogenicity and development
Abnormalities:
no effects observed
Developmental effects observed:
not specified

Table 1: Ovarian, Uterine and Litter Data

Parameter

Control

1500 mg/kg bw/day

Mean utegrine weights (g)

Gravid

69.48 ± 14.0

66.90 ± 15.2

Mean uterine weights (g)

Nongravid

6.52 ± 1.4a

7.18 ± 2.6a

Corpura lutea (mean number)

15.9

14.8

Implantations (number)

13.5

12.7

Mean implantation efficiency (indeces)

85.7a

87.1a

Resorptions (number)

1.2

0.5

Mean incidence of resorptions (%)

9.2a

4.6a

Dead fetuses (number)

0.0

0.0

Mean incidence of fetal mortality (%)

0.0a

0.0a

Live fetuses (number)

12.4 ± 2.6

12.2 ± 2.5

Mean incidence of fetal viability (%)

90.8a

95.5a

Incidences were calculated on a per litter basis.

Data are shown as means.

a: Data analysed following rank transformation

 

Table 2: Examination of the fetuses

Parameter

Control

1500 mg/kg bw/day

Live fetuses

Males

Mean body weight (g)

3.58 ± 0.201b

3.50 ± 0.47b

Mean crown-rump distance (cm)

3.8 ± 0.15b

3.7 ± 0.28b

Females

 

 

Mean body weight (g)

3.39 ± 0.186b

3.36 ± 0.45b

Mean crown-rump distance (cm)

3.7 ± 0.14b

3.6 ± 0.28b

Males per litter (Mean %)

54.0 ± 18.31

55.7 ± 14.97

Gross pathology findings:

Number of fetuses examined

191

170

Number of fetuses appearing normal

169

154

Skull

0

0

Palate

0

0

Cleft palate

0

4

Heart

0

0

Kidney

0

0

Ureter

0

0

Dilated

16

10

Hydroureter

0

1

Undulated

2

1

Umbilical hernia

1

0

Situs inversus

1

0

Hindleg

0

1

Small

1

0

Exencephaly

0

0

Tail missing

0

0

Mean incidence values for visceral findings

Number of litters

22

20

Number with anomalies

0a

0.1 ± 0.45a

Incidence of anomalies (%)c

0a

5 ± 22.4a

Number with variants

0.1 ± 0.29a

0.1 ± 0.45a

Incidence of variants (%)d

3 ± 9.7a

5 ± 22.4a

Mean incidence values for skeletal findings

Number of litters

22

20

Number with anomalies

0a

0a

Incidence of anomalies (%)e

0a

0a

Number with variants

3 ± 2.3a

3 ± 2.1a

Incidence of variants (%)f

39 ± 25.2a

40 ± 27.7a

Visceral Findings

Number of fetuses examined/number of litters

81/22

73/20

Number of fetuses with anomalies/number of litters with anomalies

0/0

2/1

Number of anomalies

0

2

Cleft palate

0

2

Number of fetuses with variants/number of litters with anomalies

2/2

2/1

Number of variants

2

2

Dilated renal pelves

2

0

Dilated ureters

0

0

Fetus small

0

2

Small tongue

0

0

Skeletal Findings

 

 

Number of fetuses examined/number of litters

191/22

170/20

Number of fetuses with anomalies/number oflitters with anomalies

0/0

0/0

Number of anomalies

0

0

Number of fetuses with variants/number of litters with variants

73/19

65/18

Number of variants

110

123

Skull:

 

 

Intraparietal-incomplete ossification/nonossified

21

19

Supraoccipital-incomplete ossification/nonossified

14

19

Hyoid

25

24

Supraoccipital - nonfused

0

1

Supraoccipital - lopsided

0

0

Supraoccipital - small

0

0

Small

0

0

Nasal bones appear short

0

0

Rib cage:

 

 

Parletals – irregular edges

2

0

Ribs – angulated

2

3

Ribs – 13thpair small

1

0

Ribs – first pair small

0

0

Ribs – forked

0

0

Vertebrae:

 

 

Centra – incomplete ossification / nonossified

32

21

Centra – nonfused

10

11

Centra small

0

0

Centra – missing

0

3

Sternabrae – bipartite

1

0

Sternebrae - less than 3 ossified

0

4

Vertebrae – less than 3 ossified

0

0

No of vertebrae below 3rdsacral

0

0

Sacral vertebrae – nonossified

0

0

Halaligned centra ans arches / caudals

0

0

Caudals – less than 3 ossified

0

1

Caudals – incomplete ossification

0

0

Arches – nonossified

0

3

Pelvis:

 

 

Pelvic girdle – incomplete ossification

1

5

Pelvic girdle – small

0

1

Pubis – incomplete ossification/nonossified

1

4

Pubis – small

0

1

Ischium – incomplete ossification / nonossified

0

3

Data are shown as mean ± standard deviations.

a: Incidences were calculated on a per litter basis.

b: Data analysed following rank transformation

c: Mean Incidence of Visceral Anomalies (%) – Group mean of ((number of fetuses with anomalies per litter/number of fetuses examined viscerally per litter)x100)

d: Mean Incidence of Visceral Variance (%) – Group mean of ((number of fetuses with variants per litter/number of fetuses examined viscerally per litter)x100)

e: Mean Incidence of Skeletal Anomalies (%) – Group mean of ((number of fetuses with anomalies per litter/number of fetuses examined skeletally per litter)x100)

f: Mean Incidence of Skeletall Variance (%) – Group mean of ((number of fetuses with variants per litter/number of fetuses examined skeletally per litter)x100)

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The available information comprises adequate, reliable studies (Klimisch score 2) and consistent studies from reference substances with similar structure and intrinsic properties. Read-across is justified based on a common functional group, common precursors/breakdown products and similarities in toxicological properties (refer to endpoint discussion for further details). The selected studies are thus sufficient to fulfil the standard information requirements set out in Annex IX, Section 8.7.2, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006. A prenatal developmental toxicity study in a second species as set out in Annex X, Section 8.7.2 does not need to be conducted and is scientifically unjustified, as no adverse effects on intrataurine development in the first species or any adverse systemic effects in acute toxicity, repeated dose toxicity, genetic toxicity and developmental toxicity studies were observed and as fatty acids fulfil physiological functions within the body.
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

Human health effects on toxicity to reproduction are predicted from adequate and reliable data for source substances by read-across to the target substance within the group applying the group concept in accordance with Annex XI, Item 1.5, of Regulation (EC) No 1907/2006.

Fatty acids are found in all living organisms fulfilling three fundamental roles. Besides their function as part of molecules like phospholipids and glycolipids important for the cell-structure, they are often precursors of signalling molecules such as prostanoids. The third and best understood role of fatty acids is their role as nutritional energy source.

Based on this, no developmental toxicity/teratogenicity is expected for fatty acids which is demonstrated by studies with C7 fatty acid (heptanoic acid), C8 fatty acid (octanoic acid), C9 fatty acid (nonanoic acid and azelaic acid), C18:2 unsaturated fatty acid (linoleic acid and conjugated linoleic acid), C22 fatty acid (docosanoic acid) and fatty acids, tall oil, respectively.

In prenatal developmental toxicity studies similar to OECD Guideline 414, groups of 22 pregnant Crl:COBS, CD® (SD) BR rats were administered via gavage 1000 mg/kg bw/day of heptanoic acid (CAS# 111-14-8) or 1500 mg/kg bw/day of nonanoic acid (CAS# 112-05-0) from day 6 through 15 of gestation (1983). A further group of 22 animals served as control group and received the vehicle corn oil.

Survival rate, body weight gain, food and water consumption were comparable between treated and control groups. Gross pathology findings were considered to be incidental in nature and showed no relationship to treatment. In addition, gravid uterus weights, the mean number of corpora lutea and implantations, mean implantation efficiency and mean incidence of resorptions revealed no evidence of maternal toxicity both in rats given heptanoic acid and nonanoic acid.

No alterations in the number of live and dead fetuses were observed. Foetal viability, size and sex remained unchanged in both groups receiving either heptanoic or nonanoic acid. In the heptanoic acid group, anomalies determined in gross pathology, visceral and skeleton examinations of pups were present in both control and treated animals and are therefore not considered as treatment-related effects. In the nonanoic group, 4 pups from the same dam receiving skeletal examinations revealed a cleft palate. Moreover a higher mean incidence of visceral anomalies was noted due to a cleft palate observed in two fetuses from the same dam. However, this increase was not statistically significant. Thus, heptanoic acid and nonanoic acid are not considered to induce maternal toxicity, developmental toxicity or teratogenicity in a concentration up to and including 1000 mg/kg bw/day and 1500 mg/kg bw/day, respectively.

The heptanoic acid and nonanoic acid doses applied in the prenatal developmental toxicity studies (1983) were determined in previous dose-range finding studies (1982). In these so called maternal tolerance studies, pregnant rats (6 females per dose) were exposed to 100, 500 and 1000 mg heptanoic acid/kg bw/day and to 200, 1000 and 2000 mg nonanoic acid/kg bw/day from gestational day 6 – 15, respectively. Criteria evaluated were mortality, clinical signs, body weights, food and water consumption and after sacrifice on gestational day 16 gross pathology, and ovarian and uterine data.

No adverse effects in groups dosed with heptanoic acid were noted compared to control. An apparent treatment related effect for nonanoic acid was a non-increase of body weight on day 9 compared to day 6 in the 2000 mg/kg bw/day group. However, the mean body weight change from gestation day 6-15 for the high-dose group was comparable to the control animals. No alterations in the number of implantations or resorptions, nor in the number of live and dead foetuses were observed between the control and the groups treated with heptanoic acid and nonanoic acid, respectively.

 

Moreover a Reproduction / Developmental Toxicity Screening Test performed equivalently to OECD 421 is available for heptanoic acid (CAS# 111-14-8) (1990). 10 female Sprague-Dawley rats were orally treated with 200, 1000 or 2000 mg/kg bw/day of the test substance dissolved in corn oil. Control animals received the vehicle only. Females were treated 7 days prior to mating and afterwards during mating, gestation and lactation until day 4 post-parturition. Male rats were used only as breeders (10 animals/group), and were not given the test substance.

The offspring was examined for numbers, viability, body weights, sex ratios and external morphology of the pups. Delivered pups were additionally observed for viability, clinical observations and body weights during a four-day post-parturition period. Afterwards, they were sacrificed and subjected to post-mortem examination of malformations and gross lesions of the thoracic and abdominal viscera.

In the offspring, there were no adverse clinical signs and no effects on viability and body weights. Gross pathology revealed no malformations or gross lesions up to and including the highest dose tested. Based on these results the NOAEL for developmental toxicity was determined to be ≥ 2000 mg/kg bw/day.

The toxicity to reproduction of docosanoic acid (CAS# 112-85-6) was evaluated in a combined repeated dose and reproductive/developmental toxicity screening test performed under GLP according to OECD guideline 422 (2002). Groups of 13 male and 13 female Sprague-Dawley rats received daily doses of 100, 300 and 1000 mg/kg bw of docosanoic acid by gavage. While the males were treated for 42 days, the females received the test substance from 14 days prior to mating until day 3 of lactation. Since the viability, the body weights, and the results of pathological examination of the pups did not reveal any differences between the dose groups and the control groups, there was no indication for any adverse effects on development. Thus, the NOAEL for developmental toxicity and teratogenicity was ≥ 1000 mg/kg bw/day based on the absence of treatment-related effects.

In a two-generation reproduction toxicity study similar to OECD Guideline 416 Sprague-Dawley rats (30 females per group and 15 males per group) were administered 5% and 10% fatty acids, tall oil (CAS# 61790-12-3) in the diet (corresponding to 2500 and 5000 mg/kg bw/day) (Pine Chemical Association, 2004). The parental animals were fed the test substance starting at 20 days pre-mating until the weaning of the first generation (F1). After weaning, 20 F1 males and 20 F1 females per group were maintained on the parental diet and mated at 100 days of age. The delivered F2 pups were followed until weaning. Haematology, serum chemistry, urinalysis, and organ weights for F1 animals, gross pathology of F1 and F2 animals and microscopic pathology of various organs of the F2 pups were performed.

There were no treatment-related effects on the number of liveborn or still-born F1 litters and pups, or weaning weight of the F1 pups. In the pups no changes in haematology, clinical chemistry and urinalysis parameters and organ weights were found and gross and microscopic pathology revealed no treatment-related effects. Thus, the NOAEL for developmental toxicity of fatty acids, tall oil was ≥ 5000 mg/kg bw/day.

There are several publications available, which also point out, that fatty acids do not have properties of inducing developmental toxicity or teratogenicity. In general, the developmental studies reported in these publications were not performed according/similar to current guidelines.

The effect of octanoic acid (CAS# 124-07-2) on foetal development was analysed in a developmental screening study with a Chernoff/Kavlock assay (Narotsky et al., 1994). Groups of 16 Sprague-Dawley rats received a daily dosage of 1125 and 1500 mg/kg bw/day in corn oil on gestation days 6 - 15 via gavage. The dose levels were based on preliminary range-finding studies with nongravid female rats. Maternal body weights were determined and the animals were observed for clinical signs throughout the experimental period ending on postnatal day 6. Uterine implantation sites were recorded. Pups in each litter were examined externally, counted and weighed. 2 pups per litter were selected on postnatal day 6 for skeletal examinations.

A maternal mortality rate of 31% and 44% was found in the low- and high-dose group. Most of the deaths occurred shortly after dosing and were attributed to the respiratory effect of the treatment. However, the high percentage of deaths could have been also due to tracheal intubation according to the authors. Maternal toxicity like rale and dyspnoea was noted in 95% of the test animals and reduced weight gain was observed in the animals of both dose groups taken into account gestation day 6 – 20. Decreased progeny viability on post-gestational day 6 occurred in dams with peripartum respiratory symptoms of the high-dose group, while all pups gained weight and no malformations including lumbar ribs were found. Based on these findings, the NOAEL for teratogenicity was determined to be ≥ 1500 mg/kg bw/day, whereas the NOAEL for developmental toxicity was considered to be 1125 mg/kg bw/day and the NOAEL for maternal toxicity was found to be below 1125 mg/kg bw/day.

Developmental toxicity of octanoic acid (CAS# 124-07-2) was also evaluated in a published study, where 12 Sprague-Dawley rats received a single dose of 18.75 mmol/kg bw by gavage on day 12 of gestation (Scott et al., 1994). As a result of the dosing, severe maternal toxicity was noted, but not further specified. The analysis of foetuses was performed on day 20 of gestation. In addition, the content of octanoic acid in foetuses and in maternal fluid was analysed 0.25, 0.5, 1, 2, 4, 8, and 24 hours after dosing. According to the authors the observed maternal toxicity was the reason for the slight reduction in the foetal body weight when compared to control values. However, no significant differences to control values were found regarding number of living foetuses, number of implantation sites, number of resorptions and number of malformations, respectively. Thus, the NOAEL for developmental toxicity was found to be ≥18.75 mmol/kg bw (total dose) corresponding to ≥2704 mg octanoic acid/kg bw.

In another study which is insufficient for assessment due to limited documentation, octanoic acid (CAS# 124-07-2) was injected subcutaneously once to mice at a dose of 600 mg/kg bw/day on day 8 of gestation (Nau and Loescher, 1986). According to the authors, no significant differences compared to control were found for octanoic acid. No further information was provided.

Developmental toxicity and teratogenicity of azelaic acid (CAS# 123-99-9) were investigated in rats and rabbits by Mingrone et al. (1983). One group of female rats and female rabbits was administered azelaic acid via feed (rat: 140 mg/kg bw/day; rabbit: 200 mg/kg bw/day) during pregnancy. Rats were sacrificed on gestation day 19 and rabbits on gestation day 25, respectively. No abnormalities were found in either the macro- or microscopic appearance of uteri, placentae and ovaries and no abortions or foetal malformations were detected in both species. The number and the weight of the foetuses were comparable to the control groups.

In a second group of female rabbits and female rats, the treatment with azelaic acid was extended for a period of 3 months, including the period of suckling before sacrifice. No significant differences were noted in the number or weight of animals born from mothers fed with azelaic diet. No still-born animals were found and new-born animals did not present any malformations. Thus a NOAEL for developmental toxicity and teratogenicity of ≥ 140 mg/kg bw/day for rats and ≥ 200 mg/kg bw/day for rabbits was determined, respectively.

Linoleic acid (CAS# 60-33-3) was evaluated in a Chernoff/Kavlock developmental toxicity screening test among 55 other test substances (Seidenberg and Becker, 1987). 26 - 30 timed-pregnant mice were administered by gavage linoleic acid on gestation days 8 – 12. The concentration was not specified in the publication, however, in general a single minimally toxic dose was used or if test substances did not produce toxicity in a preliminary range-finding study, the animals were exposed to a dose level of 10 g/kg bw/day. Since no statistically significant difference was seen in the parameters examined (average number of live neoantes per litter, average number of dead neonates per litter, average neonatal body weight) in comparison to the control group, linoleic acid was designated as nonteratogen/nonembryotoxin.

The effect of conjugated linoleic acid (CLA) on neonatal growth and development in Fischer rats was examined in two experiments by Chin et al. (1994).

In the first experiment rats were either fed a basal diet (control group) or basal diet supplemented with 0.5% CLA (corresponding to 250 mg/kg bw/day) throughout gestation (~ 20 dams per group). On day 20 of gestation, 10 rats of the treatment group and 9 rats of the control group were sacrificed and liver, mammary gland, skeletal muscle and abdominal adipose tissue were collected. Fetuses were removed, weighed and examined grossly for abnormalities. 11 further dams of the control group and 13 further dams in the CLA-fed group were allowed to deliver at term.

The CLA consumption during gestation had no effects on maternal food intake, body weight and mammary gland or liver weight. There were no significant differences in litter sizes, foetal body weights, foetal liver and brain weights between the control and CLA treated group on day 20 of gestation. Additionally, no gross visible foetal abnormality was observed. In tissues of maternal and foetal liver, maternal muscle and mammary gland, the amount of CLA was increased significantly over control levels.

Milk collection on day 10 of lactation revealed, that CLA was incorporated into milk fat of dams fed the CLA-supplemented diet. Pups receiving CLA during gestation and lactation showed significant higher mean pup weights on day 10 of lactation than pups from control dams (P < 0.036).

In the second experiment (Chin et al., 1994) Fischer rats received either a basal diet (control group) or basal diet supplemented with 0.25% or 0.5% CLA (corresponding to 125 and 250 mg/kg bw/day) throughout gestation and lactation. A further group received basal diet during gestation followed by 0.5% CLA-supplemented basal diet during lactation. Pups were weaned on day 22 of lactation and fed for 8 (males) and 10 (females) weeks, respectively, the same diet their mothers had received. The concentration of CLA in maternal milk increased in proportion to the dietary CLA level. Litter size, pup development and survival rate were not affected in any of the treatment groups. However, pup body weights were significantly increased on day 10 of lactation in the 0.5% CLA-supplemented diet group and in pups fed 0.25 and 0.5% CLA for 8 to 10 weeks of age compared to control group. Although food intake was not affected by CLA treatment for the time after weaning to 8 and 10 weeks of age, the feed efficiency was significantly increased in male pups fed 0.5% CLA and in female pups fed 0.25 and 0.5% CLA.

Based on the result of these two experiments, the NOAEL for maternal and developmental toxicity was ≥ 0.5% CLA (corresponding to ≥ 250 mg/kg bw/day).

Moreover, the effect of conjugated linoleic acid (CLA; composition: 42.6% cis-9, trans-11 CLA, 45.6% trans-10, cis-12-CLA and 8.7% other CLA isomers) on body weight gain and body composition during pre- and postnatal period was investigated in rats by Poulos et al. (2001). Sprague-Dawley rats were fed either a basal diet or the basal diet supplemented with 0.5% CLA (corresponding to 250 mg/kg bw/day) from gestation day 7 until lactation day 21. Litters were normalized to 6 pups (3 per sex) within 24 hours of birth. At weaning (day 21), one male and female pup of each litter were sacrificed and inguinal fat pads removed. The remaining progeny were assigned to control or CLA supplemented diet (each 1 male and female pup per litter) and fed until 11 weeks of age.

Maternal body weights during gestation and lactation, feed intake and liver weight of dams were not affected by CLA treatment. There were no differences in the litter size between CLA and control group. At weaning (day 21), female pups exposed to CLA were significantly heavier than female pups of dams fed control diet. Male pup weight was not influenced by CLA treatment at weaning, although males exposed to CLA throughout gestation, lactation and post-weaning were the heaviest, fastest growing and most feed efficient of all treatment groups. Dam’s diet did not alter average daily body weight gain or feed intake in either male or female pups. CLA had no effect on liver weight in either male or female pups. The main long-term effect of the dam’s diet was the significantly heavier soleus and gastrocnemius muscles (P < 0.01) and increased tail lengths (control: 18.01 cm; CLA: 18.65 cm), which was an indication of skeletal growth, of male pups whose dams were fed CLA.

The NOAEL for developmental toxicity was considered to be ≥ 250 mg/kg bw/day based on the results of the study.

Since all the members of the category show similar structural and toxicological properties the results of individual members are also valid for other fatty acids within the category. Moreover, a substance-specific adjustment of the NOAEL for the individual members of the category is not required. An overall NOAEL for all fatty acids within the category of 1000 mg/kg bw/day is chosen as “worst case” among the available key studies. In addition, heptanoic acid and docosanoic acid, which elicit a NOAEL for developmental toxicity and teratogenicity of ≥ 1000 mg/kg bw/day are representing category members with a low and the highest molecular weight.

In conclusion, the available data do not provide any evidence of developmental toxicity/teratogenicity of fatty acids and thus a prenatal developmental toxicity study in a second species is scientifically unjustified.

References

Chin, S.F. at al (1994). Conjugated Linoleic Acid Is a Growth Factor for Rats as Shown by Enhanced Weight

Gain and Improved Feed Efficiency. J. Nutr. 124 (12):2344-2349.

Narotsky, M.G. et al. (1994). Developmental Toxicity and Structure-Activity Relationships of Aliphatic Acids, Including Dose-Response Assessment of Valproic Acid in Mice and Rats. Fundamental and Applied Toxicology 22(2):251-265.

Nau, H. and Löscher, W. (1986).Pharmacologic evaluation of various metabolites and analogues of valproic acid: teratogenic potencies in mice. Fundam Appl Toxicol, May, 6(4):669-676.

Mingrone, G. et al (1983). Toxicity of azelaic acid. Drugs Exptl. Clin. Res. 9(6):447-455.

Scott, W.J. Jr et al. (1994). Pharmacokinetic Determinants of Embryotoxicity in Rats Associated with Organic Acids. Environ Health Perspect, 102, Suppl 11:97-101.

Pine Chemical Association (2004). Final Submission for Tall Oil Fatty Acids and Related Substances - VII. Robust Summaries of Data for Tall Oil Fatty Acids and Related Substances.Bibliographic source: no data

Seidenberg, J.M. and Becker, R.A. (1987) A Summary of the Results of 55 Chemicals Screened for Developmental Toxicity in Mice. Teratogenesis, carcinogenesis, and mutagenesis 7(1):17-28.

Poulos, S.P. et al. (2001). Pre- and Postnatal Dietary Conjugated Linoleic Acid Alters Adipose Development, Body Weight Gain and Body Composition in Sprague-Dawley Rats. J. Nutr. 131(10):2722-2731.

Justification for classification or non-classification

All available data on toxicity on reproduction of the members of the fatty acids category do not meet the criteria for classification according to Regulation (EC) No 1272/2008, and are therefore conclusive but not sufficient for classification.

No information is available on effects via lactation.