Fatty acids, C16-18

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Studies on reproductive toxicity by the oral route are available for the following category members:
CAS# 112-85-6, C22: NOAEL oral (fertility) = 1000 mg/kg bw/d; highest dose tested (Nagao 2002), OECD 422

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

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP-Guideline Study, tested with the source substance CAS 112-85-6. In accordance to ECHA guidance document "Practical guide 6: How to report read-across and categories (May 2010)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

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 or test system 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:

- 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:

daily

Details on study schedule:

- Age at mating of the mated animals in the study: 10 weeks

Dose / conc.:

100 mg/kg bw/day (nominal)

Dose / conc.:

300 mg/kg bw/day (nominal)

Dose / conc.:

1 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

13 in each group

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
- Cage side observations checked in table were not included.

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

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 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 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.

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 findings on fertility were noted

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

Generation:

F1

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 findings were noted

Reproductive effects observed:

not specified

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± S.D (N)

Dose group (mg/kg)	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)

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Fatty acids are found in all living organisms fulfilling fundamental physiological functions within the body. Based on this role within the body, no potential of fatty acids for affecting reproduction is expected as it could be demonstrated with fatty acids C22.

A NOAEL of 1000 mg/kg bw/d for fertility was found in a combined repeated dose toxicity study with the reproduction / developmental toxicity screening test performed according to OECD Guideline 422 with C22 fatty acid (Nagao et al., 2002). Groups of 13 male and 13 female Sprague-Dawley rats received daily doses of 100, 300 and 1000 mg/kg bw/d of docosanoic acid by gavage. While the males were treated for 42 days, the females received the test substance form 14 days prior to mating until day 3 of lactation. As result, neither mortality nor abnormalities in general condition were observed in all dose groups. Since also no difference in number of corpora lutea, implantation rate, number of implantations and all other reproductive parameters was noted in all treated groups and in the control group, there were no indications of any adverse effects on reproduction.

Short description of key information:
Studies on reproductive toxicity by the oral route are available for the following category members:
CAS# 112-85-6, C22: NOAEL oral (fertility) = 1000 mg/kg bw/d; highest dose tested (Nagao 2002), OECD 422
No data are available for reproductive toxicity after dermal exposure and inhalation, respectively.

Effects on developmental toxicity

Description of key information

Studies on developmental toxicity/teratogenicity by the oral route are available for the following category members:

CAS# 124-07-2, C8: NOAEL oral (developmental toxicity/teratogenicity) = 2704 mg/kg bw/d (Scott 1994)

CAS# 57-10-3, C16: NOAEL oral (developmental toxicity/teratogenicity) = 1500 mg/kg bw/d (Narotzky 1994)

CAS# 112-85-6, C22: NOAEL oral (developmental toxicity/teratogenicity) = 1000 mg/kg bw/d; highest dose tested (Nagao 2002), OECD 422

CAS# 124-07-2, C8: NOAEL oral (developmental toxicity/teratogenicity) = 600 mg/kg bw/d (Nau and Loescher 1986) RL4

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

Link to relevant study records

Reference

Reference 1

Endpoint:

developmental toxicity

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP-Guideline Study, tested with the source substance CAS 112-85-6. In accordance to ECHA guidance document "Practical guide 6: How to report read-across and categories (May 2010)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

other: 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

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Japan, Hino, Japan
- 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:

corn 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

Analytical verification of doses or concentrations:

yes

Details on mating procedure:

- Impregnation procedure: cohoused
- If 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

Duration of treatment / exposure:

- males: 42 days
- females: from 14 days prior to mating to day 3 of lactation

Frequency of treatment:

once daily

Duration of test:

from 14 days prior to mating to day 3 of lactation

Dose / conc.:

100 mg/kg bw/day (nominal)

Dose / conc.:

300 mg/kg bw/day (nominal)

Dose / conc.:

1 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

13 in each group

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

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least once a day
- Cage side observations checked in table were not included.

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

POST-MORTEM EXAMINATIONS: Yes
- Organs examined: brain, liver, spleen, thymus, kidney, adrenal, bladder, heart, ovary, uterus

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: No
- Number of late resorptions: No

Fetal examinations:

- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: all per litter
- Skeletal examinations: No
- Head examinations: No

Statistics:

Yates test, Mann-Whitney U test, Fisher exact test, Bartlett test, Dunnett test, Scheffe test, Kruskal-Wallis test

Indices:

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 = (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

Details on maternal toxic effects:

Details on maternal toxic effects:
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
- Females: no deaths were observed in any treatment group

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
- 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 group. However, since no other changes in food consumptio was noted in 300 mg/kg and 1000 mg/kg, it was considered not to be related to the dosing of compound.

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)
- Females: kidney weight was significantly reduced 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)
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 tubules of the cortex 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.

Basis for effect level:

other: maternal toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects. Remark: no adverse substance -related findings were noted

Details on embryotoxic / teratogenic effects:
VIABILITY (OFFSPRING)
No change in numbers of birth, birth rate, infant live birth rate, infant survival and birth rates at day 4 was found in all dose groups when compared to the values of the control group.

BODY WEIGHT (OFFSPRING)
No difference in body weight 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

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 findings were noted

Abnormalities:

no effects observed

Developmental 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 ± S.D. (N)

Dose group (mg/kg)	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)

Reference 2

Endpoint:

developmental toxicity

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well documented publication which meets basic scientific principles

Principles of method if other than guideline:

Screening for developmental toxicity with a Chernoff/Kavlock assay.

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Route of administration:

oral: gavage

Vehicle:

corn oil

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

on gestation days 6-15

Frequency of treatment:

once daily

Duration of test:

28 d

Dose / conc.:

1 125 mg/kg bw/day (nominal)

Dose / conc.:

1 500 mg/kg bw/day (nominal)

No. of animals per sex per dose:

16

Control animals:

yes, concurrent vehicle

Dose descriptor:

NOAEL

Effect level:

< 11 250 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Dose descriptor:

NOAEL

Effect level:

1 500 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: teratogenicity

Abnormalities:

not specified

Developmental effects observed:

not specified

Sprague Dawley rats (n=16/group) gavaged on gestation days 6-15 with caprylic acid in corn

oil at dose levels of 0, 1 125 or 1 500 mg/kg bw/day exhibited maternal toxicity (rale and

dyspnoea) and maternal mortality (21 % and 44 % respectively). Most deaths occurred shortly

after dosing and were attributed to the respiratory effect of the treatment. However, according

to the authors, the high percentage of deaths could have been due to tracheal intubation. There

was a decrease in the number of live pups on post-gestational day 6, while no teratogenic

effect was observed

Reference 3

Endpoint:

developmental toxicity

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Basic data given

Qualifier:

no guideline followed

Principles of method if other than guideline:

Developmental toxicity study with rats, which reveived a single dose of 18.75 mmol/kg bw octanoic acid by gavage on day 12 of gestation. Fetuses were analyzed on day 20 of gestation.

GLP compliance:

no

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (Portage, MI).
- Housing: in small groups in hanging wire cages
- Diet (e.g. ad libitum): Purina Rodent Laboratory Chow; ad libitum
- Water (e.g. ad libitum): water; ad libitum
- Acclimation period: 5 - 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 - 23
- Humidity (%): 45 - 55
- Photoperiod (hrs dark / hrs light): 12 / 12

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

- Impregnation procedure: purchased timed pregnant
- Proof of pregnancy: vaginal plug referred to as day 0 of pregnancy

Duration of treatment / exposure:

single dosing

Frequency of treatment:

single dosing

Duration of test:

9 days (from day 12 to day 20 of gestation)

Dose / conc.:

18.75 other: 18.75 mmol/kg bw (nominal)

Dose / conc.:

2 704 mg/kg bw/day

Remarks:

Basis: nominal in water (calculated)

No. of animals per sex per dose:

Octanoic acid: 12
Control: 10

Control animals:

yes, concurrent vehicle

Maternal examinations:

CAGE SIDE OBSERVATIONS: No

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: No:

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

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day #20
- Organs examined: Uterus

OTHER
- Pharmacokinetics: Content of octanoic acid inmaternal plasma at 0.25, 0.5, 1, 2, 4, 8, and 24 h after dosing

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: No
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes

Fetal examinations:

- External examinations: Yes: 1/3 per litter
- Soft tissue examinations: Yes: 2/3 per litter
- Skeletal examinations: Yes: 2/3 per litter
- Head examinations: Yes: 2/3 per litter
-Other: Content of octanoic acid in embryo at 0.25, 0.5, 1, 2, 4, 8, and 24 h after dosing

Details on maternal toxic effects:

Details on maternal toxic effects:
Severe maternal toxicity was noted (not specified)

Dose descriptor:

NOAEL

Effect level:

2 704 mg/kg bw (total dose)

Based on:

test mat.

Basis for effect level:

other: developmental toxicity

Details on embryotoxic / teratogenic effects:

Details on embryotoxic / teratogenic effects:
Octanoic acid was essentially devoid of embryotoxic effects except for a slight reduction of fetal weight, most likely attributable to the severe maternal toxicity that accompanied the administration of this agent.

Abnormalities:

not specified

Developmental effects observed:

not specified

Results on embryotoxicity:

	Octanoic acid	Control
Dose (mg/kg bw)	2704	0
No. surv. females/no. Treated	11/12	10/10
No. impl. Sites	131	108
No. res. or dead (%)	10 (9)	6 (6)
No. surv. malf. (%)	4 (3)	1 (1)
Mean fetal wt., M/F	3.48/3.23	3.90/3.71

Weight of day 20 rat fetuses exposed to an organic acid on day 12 of pregnancy:

	Control		Octanoic acid
Litter No.	Male	Female	Male	Female
1	3.6	3.4	3.8	3.7
2	4.2	3.9	3.8	3.5
3	4	3.7	2.5	2.2
4	4	3.9	3.7	3.3
5	4	3.8	3.5	3.2
6	3.9	3.7	3.3	3.2
7	3.5	3.4	3.8	3.7
8	4.3	4.1	4.1	3.9
9	4.5	4.2	2.2	2
10	3.9	3.6	-	-
mean	3.99 ± 0.30	3.77 ± 0.27	3.19 ± 0.65	3.19 ± 0.66
% variability	7.5	7.2	19.1	20.7

Transplacental Pharmacokinetics:

Based on the data presented in figures, ca. 70 µg/ml octanoic acid was found in maternal plasma and ca. 200 µg/g fetus directly after administration, respectively. While octanoic acid could not be detected anymore in maternal plasma after ca. 2 hours, it took 5 hours until octanoic acid could not be detected anymore within the embry.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

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

Fatty acids are found in all living organisms fulfilling fundamental physiological functions within the body. Based on this role within the body, no potential of fatty acids for affecting development is expected as it could be demonstrated with C8 fatty acid (octanoic acid), C16 fatty acid (palmitic acid) and C22 fatty acid (docosanoic acid), respectively.

Developmental toxicity by octanoic acid was evaluated in a published study, where 12 Sprague-Dawley rats received a single dosage 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 fetuses was performed on day 20 of gestation. In addition, the content of octanoic acid in fetuses and in maternal fluid was analyzed 0.25, 0.5, 1, 2, 4, 8, and 24 hours after dosing. According to the author, the observed maternal toxicity was the reason for the slight reduction in the fetal body weight when compared to control values. However, no significant differences to control values were found regarding number of living fetuses, 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/kg bw octanoic acid.

The effect of palmitic acid on development was analyzed in a 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/d in corn oil on gestation days 6 - 15 via gavage. A maternal mortality rate of 21% and 44% was found where ,according to the authors, most of the deaths could have been attributed to intubation error. However, maternal toxicity like rale and dyspnea was noted. There was a decrease in the number of live pups on post-gestational day 6, while no teratogenic effect was observed. Based on these findings, the NOAEL for teratogenicity is 1500 mg/kg bw/d, whereas the NOAEL for maternal toxicity was found to be below 1125 mg/kg bw/d.

A NOAEL of 1000 mg/kg bw/d was found for docosanoic acid in a combined repeated dose toxicity study with the reproduction/developmental toxicity screening test performed according to OECD Guideline 422 with C22 fatty acid (Nagao et al., 2002). Groups of 13 male and 13 female Sprague-Dawley rats received daily doses of 100, 300 and 1000 mg/kg bw/d of docosanoic acid by gavage. While the males were treated for 42 consecutive days, the females received the test substance form 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.

In another study which is insufficient for assessment due to limited documentation, octanoic acid was injected subcutaneously once to mice at a dose of 600 mg/kg bw/d 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.

Taking all the results together, the study data do not provide any evidence of developmental toxicity/teratogenicity of fatty acids which is supported by the physiological function of fatty acids within the body.

Justification for classification or non-classification

According to CLP (1272/2008/EC) classification criteria for toxicity to reproduction/developmental toxicity/teratogenicity, fatty acids do not fulfill the criteria for classification and thus a non-classification is warranted for this endpoint.