Fatty acids, C16-18 (even numbered), ammonium salts

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Toxicity to reproduction (fertility): Based on the experimental results on analogue substance, the weight of evidence approache was applied and the NOAEL for toxicity to reproduction (fertility) for the substance Fatty acids, C16-18 (even numbered), ammonium salts, was determined to be 1000 mg/kg bw/day based on the worst case assumption.

Extended one-generation reproductive toxicity study: The study does not need to be conducted since based on the available data from the reproduction/developmental toxicity screening test and the 28 and 90-day toxicity studies, the test material did not cause any adverse effect on reproductive organs and tissues.

Link to relevant study records

Reference

Reference 1

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Crj: CD(SD)

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: 312.1-363.7 g males; 205.3-230.8 g females
- 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 ºC
- Humidity (%): 50-65 %
- Air changes (per hr): 15 per hour
- Photoperiod (hrs dark / hrs light): 12 hours dark / 12 hours light

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
Solutions prepared more frequently than once a week. Aliquots kept refrigerated in airtight conditions.

VEHICLE
- Justification for use and choice of vehicle (if other than water): Insoluble 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

Duration of treatment / exposure:

Exposure duration:
Males, 42 days
Females, from 14 days prior to mating to day 3 of lactation

Frequency of treatment:

Daily

Dose / conc.:

0 mg/kg bw/day (actual dose received)

Remarks:

vehicle

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 animals per sex and dose.

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:

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Once a day

BODY WEIGHT: Yes
- Time schedule for examinations: once a week

FOOD CONSUMPTION:
- Time schedule for examinations: once a week

HAEMATOLOGY: Yes
- Time schedule for collection of blood: after 42 days, prior to sacrifice
- 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), platlet count, hemoglobin (Hb), hematocrit (Ht), mean corpuscular volume (MCV),mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), differentiation of leukocytes (band neutrophil, segmented neutrophil, eosinophil, basophil, monocyte, lymphocyte).

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: after 42 days, prior to sacrifice
- Animals fasted: 18 - 24 hours before sacrifice
- How many animals: all surviving animals
- Parameters checked: total protein, albumin, A/G, blood urea nitrogen (BUN), creatinine, glucose, total cholesterol, total bilirubin, triglyceride, sodium (Na), potassium (K), chloride (Cl), calcium (Ca), inorganic phosphorus (IP), alkaline phosphatase (ALP), GPT, GOT, γ-GTP.

Litter observations:

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
Clinical observations (once a day), body weight on day 0 and 4 of lactation.

DEVELOPMENTAL PARAMETERS:
Day 0 of lactation: number of pups born, delivery index, number of live pups, birth index, live birth index, sex ratio.
Day 4 of lactation: number of live pups, viability index, sex ratio.

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals, on the day after the administration period.
- Maternal animals: All surviving animals, on the day 4 of lactation. Females with no delivery were killed 4 days after the delivery expected date. Females with no copulation were sacrificed at the termination of mating period.

GROSS NECROPSY
- Organs examined: Males: tyroid gland, thymus, lung, liver, spleen. Females: thymus, spleen, adreanl gland, kidney.
- Organs weights: Males: heart, liver, kidneys, thymus, testes, epididymides. Females: heart, liver, kideny and thymus.

HISTOPATHOLOGY / ORGAN WEIGHTS
- Organs examined: Males: brain, heart, liver, spleen, thymus, kidney, urinary bladder, testis, epididymis. Females: brain, heart, liver, thymus, kidney, urinary bladder, adrenal gland, ovary.

REPRODUCTION PERFORMANCE:
Number of mated pairs, number of copulated pairs, copulation index, number of pregnant females, fertility index, pairing days until copulation and frequency of vaginal estrus.

DEVELOPMENTAL PARAMETERS:
Number of pregnant females, number of pregnant females with live pups, gestation index, gestation length in days, number of corporea lutea, number of implantation sites, implantation index.

Postmortem examinations (offspring):

GROSS NECROPSY
- Full macroscopic examinations on all of pups (external and visceral observation).

Statistics:

Dunnett’s or Scheffe’s test for continuous data, Chi square test for copulated index and fertility index, and Mann-Whitney U test or Fisher’s test for histopathological examination data.

Reproductive indices:

Copulation index, fertility index, implantation index, gestation index.

Offspring viability indices:

Delivery index, birth index, live birth index, viability index, sex ratio.

Clinical signs:

no effects observed

Description (incidence and severity):

No abnormalities in general condition were observed in any of the treated groups.

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Description (incidence):

No deaths were observed in any of the treated groups.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

There were no changes related to the dosing of compound in body weight gain.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There were no changes related to the dosing of compound in food consumption.

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Description (incidence and severity):

No adverse effects were found in hematological examinations.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

No adverse effects were found in biochemical examinations.

Urinalysis findings:

not examined

Behaviour (functional findings):

not specified

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

At autopsy following treatment for 42 days in males and on postpartum day 4 in females, no adverse effects were found in the histopathological examinations.

Histopathological findings: neoplastic:

not specified

Other effects:

no effects observed

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Description (incidence and severity):

The compound showed no adverse effects on copulation or fertility.
No changes related to the dosing of compound were observed in gestation length, gestation index, delivery and lactation.
The compound did not demonstrate any adverse effects on the sex ratio of pups.

Key result

Dose descriptor:

NOEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No treatment-related effects observed at the highest dose

Key result

Critical effects observed:

no

Clinical signs:

no effects observed

Dermal irritation (if dermal study):

not examined

Mortality / viability:

no mortality observed

Description (incidence and severity):

The compound did not demonstrate any adverse effects on viability of pups.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

The compound did not demonstrate any adverse effects on body weights of pups.

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Description (incidence and severity):

No morphological abnormalities in pups were observed in any of the treated groups.

Histopathological findings:

not examined

Other effects:

no effects observed

Description (incidence and severity):

The compound did not demonstrate any adverse effects on the sex ratio of pups.

Behaviour (functional findings):

not examined

Developmental immunotoxicity:

not examined

Key result

Dose descriptor:

NOEL

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 treatment-related effects observed at the highest dose.

Key result

Critical effects observed:

no

Key result

Reproductive effects observed:

no

Conclusions:

The NOEL of docosanoic acid for maternal and reproductive toxicity was 1000 mg/kg/day both in males and females.

Executive summary:

A combined repeated dose and reproduction/developmental screening was performed on docosanoic acid according to OECD Guideline 422. 13 rats per sex and per dose were exposed to 0 (vehicle), 100, 300 and 1000 mg/kg bw/day, males for 42 days and females from 14 days prior to mating to day 3 of lactation. The compound showed no adverse effects on copulation or fertility. No changes related to the dosing of compound were observed in gestation length, gestation index, delivery and lactation. The compound did not demonstrate any adverse effects on the sex ratio, body weights or viability of pups. Also, no morphological abnormalities in pups were observed in any of the treated groups. Based on these results the NOEL for maternal and reproductive toxicity was determined to be 1000 mg/kg bw/day for both males and females.

Reference 2

Endpoint:

one-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

A scientific study. No GLP.

Principles of method if other than guideline:

-Principle of test: From 2 to 4.5 months of age, 80 crossbred gilts were reared in a conventional grower unit where they were naturally exposed to mycoplasmal and bacterial pathogens that cause pneumonia and atrophic rhinitis. At 4.5 months of age, gilts were moved to environmentally regulated rooms (4.9 x 7.3 m) and assigned at random to 1 of 2 treatment groups: low aerial concentration of ammonia (4 to 12 ppm; mean, 7 ppm) or moderate aerial concentration of ammonia (26 to 45 ppm, mean, 35 ppm). Low concentration of ammonia was obtained by flushing of manure pits weekly, whereas moderate concentration of ammonia was maintained by adding anhydrous ammonia to manure pits that were not flushed. Mean body weight gain was measured biweekly. After 6 weeks in these environments, 20 gilts from each treatment group were slaughtered, and prevalence and severity of lung lesions and snout grades were determined.

GLP compliance:

no

Limit test:

no

Species:

pig

Strain:

not specified

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 4.5 wks

Route of administration:

inhalation

Type of inhalation exposure (if applicable):

whole body

Vehicle:

not specified

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

Female pigs were continuously exposed to 7 and 35 ppm ammonia for 6 weeks before breeding until 30 day of gestation.

Dose / conc.:

7 ppm (nominal)

Remarks:

(inhalation)

Dose / conc.:

35 ppm (nominal)

Remarks:

(inhalation)

Control animals:

no

Clinical signs:

not specified

Dermal irritation (if dermal study):

not examined

Mortality:

not specified

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

Mean daily gain (MDG) was less (P < 0.01) for gilts exposed to moderate concentration of ammonia than for gilts exposed to low concentration of ammonia after 2 weeks in their respective environments. By 4 and 6 weeks, however, MDG was similar between the 2 treatment groups.

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

Description (incidence and severity):

No statistically significant differences in age at puberty, number of live fetuses, or -to-corpus luteum ration compared to pigs exposed only to 7 ppm.

Key result

Dose descriptor:

NOAEC

Effect level:

35 ppm (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

reproductive performance

Key result

Critical effects observed:

no

Remarks on result:

not measured/tested

Key result

Reproductive effects observed:

no

No statistically significant differences were noted in ovarian or uterine weights of pigs exposed to about 7 and 35 ppm ammonia for 6 weeks. Female pigs were continuously exposed to 7 and 35 ppm ammonia for 6 weeks before breeding and until 30 day of gestation had. No statistically significant differences in age at puberty, number of live fetuses, or -to-corpus luteum ration compared to pigs exposed only to 7 ppm. No unexposed controls were included in the study.

Conclusions:

The NOAEC for parental reproductive effects of ammonia in pigs was determined to be 35 ppm by inhalation.

Executive summary:

A one generation test was performed on ammonia in pigs. At 4.5 months of age, gilts were moved to environmentally regulated rooms (4.9 x 7.3 m) and assigned at random to 1 of 2 treatment groups: low aerial concentration of ammonia (4 to 12 ppm; mean, 7 ppm) or moderate aerial concentration of ammonia (26 to 45 ppm, mean, 35 ppm). Gilts were weighed biweekly. Mean daily gain (MDG) was less (P < 0.01) for gilts exposed to moderate concentration of ammonia than for gilts exposed to low concentration of ammonia after 2 weeks in their respective environments. By 4 and 6 weeks, however, MDG was similar between the 2 treatment groups. No statistically significant differences were noted in ovarian or uterine weights of pigs exposed to about 7 and 35 ppm ammonia for 6 weeks. Female pigs were continuously exposed to 7 and 35 ppm ammonia for 6 weeks before breeding and until 30 day of gestation had. No statistically significant differences in age at puberty, number of live fetuses, or -to-corpus luteum ration compared to pigs exposed only to 7 ppm. No unexposed controls were included in the study. The NOAEC for parental reproductive effects of ammonia in pigs was determined to be 35 ppm by inhalation.

Reference 3

Endpoint:

fertility, other

Remarks:

(Repeated Dose 90-Day Oral Toxicity in Rodents)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

other: OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: SPF F344/DuCri rats from Charles River Nippon.
- Age at study initiation: (P) 5 week old
- Diet (e.g. ad libitum): Animals were maintained on CRF-1 powder diet.
- Acclimation period: 1 week
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 ± 1 ºC
- Humidity (%): 55 ± 5 %
- Air changes: 18 air changes/day
- Photoperiod (hrs dark / hrs light): 12 hour light/dark cycle.

Route of administration:

oral: feed

Vehicle:

not specified

Details on exposure:

The animals were fed CRF-1 powder diet containing concentrations of 0, 0.38, 0.75, 1.5, and 3.0% of ammonium sulfate.

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

Continuosly

Dose / conc.:

0.38 other: % (nominal in diet)

Remarks:

(equivalent to 222 mg/kg bw/day in males and 239 mg/kg bw/day in females)

Dose / conc.:

0.75 other: % (nominal in diet)

Remarks:

(equivalent to 441 mg/kg bw/day in males and 484 mg/kg bw/day in females)

Dose / conc.:

1.5 other: % (nominal in diet)

Remarks:

(equivalent to 886 mg/kg bw/day in males and 961 mg/kg bw/day in females)

Dose / conc.:

3 other: % (nominal in diet)

Remarks:

(equivalent to 792 mg/kg bw/day in males and 1975 mg/kg bw/day in females)

No. of animals per sex per dose:

10 animals per sex and per dose.

Control animals:

yes, concurrent no treatment

Details on study design:

The dose levels were set on the basis of results from a previous 2-week study with a dose level of 5% .

Parental animals: Observations and examinations:

CINICAL CHEMISTRY, HEMATOLOGY:
The following clinical parameters were determined: total protein (TP), albumin/globulin (A/G), albumin, total cholesterin, BUN, Na, Cl, K, Ca, P, GOT, GPT, alkaline phosphatase. Hematology included red blood cell count (RBC), hemoglobin (Hb), hematocrit (Hk), mean corpuscular volume (MCV), mean erythrocyte hemoglobin (MCH), mean erythrocyte hemoglobin concentration (MCHC), platelet count, and leukocyte count.

Postmortem examinations (parental animals):

ORGANS WEIGHED:
At necropsy, the following organs were weighed and absolute and relative organ weights determined: Males: brain, lung, heart, spleen, liver, adrenal, kidney, testis; Females: brain, lung, heart, spleen, liver, adrenal, kidney.

ORGANS EXAMINED HISTOPATHOLOGICALLY:
Males: brain, lung, heart, spleen, liver, adrenal, kidney, testis.
Females: brain, lung, heart, spleen, liver, adrenal, kidney.

Organs were fixed in formalin and hematoxylin-eosin slides were prepared and examined histologically.

Statistics:

Bartlett and Kruskal - wallis tests, and parametric Dunnett and Scheffe tests.

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Diarrhea was observed in males at 3% during the exposure period.

Dermal irritation (if dermal study):

not examined

Mortality:

not specified

Body weight and weight changes:

no effects observed

Description (incidence and severity):

At study end final body weights were in males 298, 273, 287, 282 and 284 g for the 0, 0.38, 0.75, 1.5 and 3% groups, respectively. In females, final body weights were 151, 157, 152, 161 and 158 g for the 0, 0.38, 0.75, 1.5 and 3% groups, respectively.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

14.2, 14.0, 14.3, 14.1, 13.8 g/rat/day in the males of the 0, 0.38, 0.75, 1.5 and 3% groups, respectively. In females, the values were 9.2, 9.1, 9.3, 9.3 and 8.4 g/rat/day for the 0, 0.38, 0.75, 1.5 and 3% groups, respectively.

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Description (incidence and severity):

No biologically significant changes were observed in any of the investigated parameters. Though there were statistical differences in some of the parameters, there was no consistent dose-effect relationship and/or all values were within the normal ranges of values normally found in the rat strain used in this study. In particular, there were no signs indicative of a metabolic acidosis.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

No biologically significant changes were observed in any of the investigated parameters. Though there were statistical differences in some of the parameters, there was no consistent dose-effect relationship and/or all values were within the normal ranges of values normally found in the rat strain used in this study.

Urinalysis findings:

not examined

Behaviour (functional findings):

not specified

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

No significant pathological effects were found. In the 3% male group myofibrosis cordis, basophilic kidney tubulus as well as spleenic melanosis were observed; in the female group basophilic kidney tubulus as well as splenic melanosis were observed. However the rate of occurrence was similar to controls.

Histopathological findings: neoplastic:

not examined

Other effects:

no effects observed

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

not examined

Key result

Dose descriptor:

NOAEL

Effect level:

886 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: overall effects

Key result

Dose descriptor:

NOAEL

Effect level:

1 975 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: overall effects

Key result

Dose descriptor:

NOAEL

Effect level:

1 792 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: effects on reproductive organs

Key result

Critical effects observed:

no

Remarks on result:

not measured/tested

Reproductive effects observed:

not specified

The NOAEL was judged to be 886 mg/kg bw/day in males and 1975 mg/kg bw/day in females. The NOAEL for reprotox is determined to be 1792 and 1975 mg/kg bw/day for males and females respectively.

Conclusions:

The NOAEL was judged to be 886 mg/kg bw/day in males and 1975 mg/kg bw/day in females. The NOAEL for reprotox is determined to be 1792 and 1975 mg/kg bw/day for males and females respectively.

Executive summary:

Fischer 344 rats (10 rats/sex/dose) were exposed during 13 weeks to diets containing 0, 0.38, 0.75, 1.5 or 3 % ammonium sulfate (corresponding to 0, 222, 441, 886, 1792 mg/kg bw/day in males and to 0, 239, 484, 961, 1975 mg/kg bw/day in females). Diarrhea was observed in males at 3% during the exposure period. Increased kidney weights in males and females at 3% and increased liver weights in females at 3% were not accompanied by histopathological changes. The relative testes weight was significantly increased at all doses, but no histological effects were found. No effect was observed on body weight, food consumption, hematological and clinical parameters. No effects on the reproductive organs were observed. The NOAEL for repeated dose toxicity was determined to be 886 mg/kg bw/day for males and 1975 mg/kg bw/day for females.

Since no adverse effects were seen on reproductive organs, the NOAEL for reprotox, is determined to be 1792 and 1975 mg/kg bw/day for males and females respectively.

Reference 4

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:

the extended one-generation reproductive toxicity study does not need to be conducted because there are no results from available repeated dose toxicity studies that indicate adverse effects on reproductive organs or tissues, or reveal other concerns in relation with reproductive toxicity

Justification for type of information:

JUSTIFICATION FOR DATA WAIVING
Extended one-generation reproductive toxicity study does not need to be conducted since based on the available data from the reproduction/developmental toxicity screening test and the 28 and 90-day toxicity studies, the test material did not cause any adverse effect on reproductive organs and tissues.

Reproductive effects observed:

not specified

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

Three studies were available. The Klimisch scores were K1 (GLP), K2 and K4. A weight of evidence approach was applied and the overall quality of the database was determined as appropriate for assessment.

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Weight of Evidence Approach (see rationale attached in IUCLID5 Section 13):

A combined repeated dose and reproduction/developmental screening was performed on docosanoic acid by the Ministry of Health, Labour and Welfare (Japan, 1998) according to OECD Guideline 422. Rats were orally exposed up to 1000 mg/kg bw/day, males for 42 days and females from 14 days prior to mating to day 3 of lactation. The compound showed no adverse effects on copulation or fertility. No changes related to the dosing of compound were observed in gestation length, gestation index, delivery and lactation. The compound did not demonstrate any adverse effects on the sex ratio, body weights or viability of pups. Also, no morphological abnormalities in pups were observed in any of the treated groups. The NOEL for maternal and reproductive toxicity was determined to be 1000 mg/kg bw/day for both males and females.

A one generation test was performed by Diekman MA et al. (1993) on ammonia in pigs. At 4.5 months of age, gilts were exposed to a low (mean 7 ppm) or to a moderate (mean 35 ppm) aerial concentration of ammonia (environmentally regulated rooms). No statistically significant differences were noted in ovarian or uterine weights of pigs exposed to about 7 and 35 ppm ammonia for 6 weeks. Female pigs were continuously exposed to 7 and 35 ppm ammonia for 6 weeks before breeding and until 30 day of gestation had. No statistically significant differences in age at puberty, number of live fetuses, or-to-corpus luteum ration compared to pigs exposed only to 7 ppm. The NOAEC for parental reproductive effects of ammonia in pigs was determined to be 35 ppm by inhalation.

In a repeated dose toxicity study (test method similar to OECD 408) reported by Takagi H et al. (1999), rats were exposed during 13 weeks to diets containing up to 3 % ammonium sulfate (corresponding up to 1792 mg/kg bw/day in males and up to 1975 mg/kg bw/day in females). The relative testes weight was significantly increased at all doses, but no histological effects were found. No effects on the reproductive organs were observed. Therefore, the NOAEL for reproductive organs was determined to be 1792 mg/kg bw/day for males and 1975 mg/kg bw/day for females.

Taking into account the available experimental results, the weight of evidence approach was applied and the NOAEL for toxicity to reproduction (fertility) for the substance Fatty acids, C16 -18 (even numbered), ammonium salts was determined to be 1000 mg/kg bw/day (based on the worst case assumption).

Extended one-generation reproductive toxicity study: The study does not need to be conducted since based on the available data from the reproduction/developmental toxicity screening test and the 28 and 90-day toxicity studies, the test material did not cause any adverse effect on reproductive organs and tissues.

Justification for selection of Effect on fertility via oral route:

The lowest NOAEL was chosen based on the worst case assumption.

Effects on developmental toxicity

Description of key information

Key study: OECD Guideline 414. GLP study. The NOAEL for maternal toxicity was determined to be 181.8 mg/kg bw/day based on decrease of food consumption and decrease of body weight gain, transient respiratory murmurs and a problable death of one female rat. The test item did not cause adverse effects in fetuses, therefore the NOAEL for fetal toxicity was determined to be 1000 mg/kg bw/day.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

13 June 2017 - 26 March 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Wistar

Remarks:

Cmdb: WI; outbred

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Experimental Medicine Centre at the Medical University in Białystok
- Age at study initiation: 12 weeks (range finding ), 13 weeks (main test)
- Weight at study initiation: 229.2 g (range finding), 246.3 g (main test). Body weights were within ±20% of the average value.
- Housing: At mating one female was placed in one cage with one male. At pregnancy the females were housed individually in plastic cages with metal wire lid and dimensions: 58 x 37 x 21 cm (length x width x height).
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: 3 weeks (range finding), 2 weeks (main test).

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-25
- Humidity (%): 41-80
- Air changes (per hr): 10-20
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

VEHICLE
- Amount of vehicle (if gavage): 0.5 mL /100 g b.w.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The samples of the test item dispersions in distilled water of each dose from four time points during the main study were chemically analyzed. Samples were collected at the beginning of the experiment, two times during the main study and at the end- at the last day of dosing animals of all three dose levels. The samples were analysed by a validated HPLC coupled with UV-DAD method.

Details on mating procedure:

- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1/1
- Length of cohabitation: 1 day (range finding), 1-3 days (main test)
- Verification of same strain and source of both sexes: Pregnant females were obtained by mating with not related males.
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy

Duration of treatment / exposure:

From 5th to 19th day of gestation

Frequency of treatment:

Once a day

Dose / conc.:

33.1 mg/kg bw/day (actual dose received)

Remarks:

Group 1

Dose / conc.:

181.8 mg/kg bw/day (actual dose received)

Remarks:

Group 2

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

Remarks:

Group 3

No. of animals per sex per dose:

25

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: A range finding study was performed.
Doses: 0 (control), 57.2, 229 and 916 mg/kg bw/day.
Number of females per group: 7.
Methodology: The same as in the main test.
On the basis of DRF study results doses for the main study were selected. No significant maternal toxic effects occurred in the highest dose of DRF study (except volvulus, which was not necessarily caused by the test item action), slight changes as decrease in food consumption between the 5th and 11th day of gestation occurred. Changes in fetuses were a decrease of weight of fetuses with placenta and fetal membranes and a decrease of weight of placenta itself. Therefore, the highest dose for the main study was maintained at similar level as 1000 mg/kg b.w. Since at the lowest dose changes as increased weight of placenta and decreased length of fetuses (with and without tail) occurred and the lowest dose should not produce evidence of toxicity, the lowest dose for the main study was decreased to 33.1 mg/kg b.w. The middle dose was 181.8 mg/kg b.w. to obtain a separation factor of 5.5.

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice a day during labour week and once a day on days off.
- Parameters: Observation for morbidity and mortality.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Day o and then 5th, 8th, 11th, 14th, 17th and 20th day of gestation.
- Parameters: Skin changes, coat changes, changes in eyes and mucous membranes,
respiratory system, circulatory system, nervous system, somatic activity and behavior.

BODY WEIGHT: Yes
- Time schedule for examinations: on 0, 2nd, 5th, 8th, 11th, 14th, 17th and 20th day of gestation.

FOOD CONSUMPTION: Yes
- Time schedule for examinations: Controlled on days of their weighing
- 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: Yes

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20 of gestation.
- Organs examined:

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes

Fetal examinations:

- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter (formation of particular cavities and presence of internal organs was evaluated)
- Skeletal examinations: Yes: half per litter (skull bones, back-bone, ribs, acromial and pelvic girdles with limbs was evaluated. Points of ossification in sternum, metacarpus of fore limbs and metatarsus of hind limbs were counted during evaluation).
- Head examinations: Yes: all per litter
- Other: Sex of each fetus, body weight, length with tail and without tail was determined. Weight of the fetuses with placenta and fetal membranes was determined first and then weight of the fetuses without placenta and fetal membranes as well as weight of placenta were measured. During the gross evaluation attention was paid to: reactions to tactile impulses, formation of body coverings, formation of limbs, number of fingers and toes, shape of head, formation of auricular conchas, presence of nasal apertures, formation of oral cavity and presence of anus and tail.

Statistics:

The obtained results were elaborated statistically with the use of STATISTICA 10, with p ≤ 0.05. The results obtained in the treated groups (group 1, group 2, group 3) were compared with the control group (group 0). For each quantitative parameter, the basic characteristics of descriptive statistics were determined: mean, standard deviation, variance and using the Shapiro-Wilk test, it was assessed whether a given parameter had a normal distribution. Then, the following statistical tests were used to assess intergroup relationships:
• for quantitative parameters having normal distribution: a one-way analysis of variance, preceded by Bartlett's test, checking the homogeneity of variance, and then a post-hoc test: Dunnett's test.
• for quantitative parameters having a non-normal distribution or in the case of heterogeneity of variance: Kruskal-Wallis test, if necessary confirmed with Dunnett’s test.

Indices:

Body weight of pregnant females, food consumption of pregnant females, number of corpora lutea, number of fetuses in litter, number of males and females in litter, weight of uteri, weight of fetuses, length of fetuses, number of ossification points in sternum and limbs.

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Single cases of coat thinning (in one female per dose in each treatment group) and alopecia (in one female in group 1) occurred only in treated groups. The frequency of this change did not increase with the increase of the dose. Furthermore, the single cases of coat thinning or alopecia were observed in control groups of other studies performed in the laboratory, so it should not be connected with the test item.
The influence of the test item on treated animals was observed at the highest dose. Twelve cases of transient respiratory murmurs were observed in group 3 and there were no similar changes in lower doses and in the control group.

Mortality:

mortality observed, treatment-related

Description (incidence):

One female of group 3 died at day 9. All remaining females in the main study survived the entire period of the experiment. Clinical changes as distinct decrease of locomotor activity, dejection, bristled coat and difficult respiration, which were observed in this female before death were not observed in the lower doses and in the control group and seem to be connected with the test item. All these changes in the highest dose of 1000.0 mg/kg b.w. could be seen as the influence of the test item on rat females.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

During the entire period of the experiment average body weight of the females of group 1 and group 2 did not differ statistically significantly from average body weight of the females of the control group. The average body weight of the females of group 3 was statistically significantly lower in comparison with the control group in 14th day of gestation.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

During the experiment, food consumption of the females in group 1 and group 2 was not statistically significantly different from the control group. Food consumption of the females in group 3 was statistically significantly lower in comparison with the control group from 5th to 20th gestation day.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

No pathological changes were stated at gross examination of the pregnant females from group 0,1, 2 and 3 performed at caesarean section.
In three females found out to be non-pregnant (1 in control group, 1 in group 1 and 1 in group 2) the presence of clear fluid in uterus was stated, which is normal during proestrus phase of estrous cycle in female rats. In one non-pregnant female from group 1 the presence of a follicular cyst in the ovary was stated, what could be the cause of ineffective mating of this female, due to the possibility of hormonal disturbances affecting ovulation and hence fertility.
Post- mortem examiniation of one female from group 3, which died in 9th day of the experiment and was not pregnant revealed following findings:
- lungs enlarged, with emphysema and foci of congestion
- digestive tract filled with gas,
- congestion of intestines, liver and kidneys.

Number of abortions:

no effects observed

Description (incidence and severity):

Fertility was equal in all groups, including control group, 23 pregnant females per group were obtained.

Pre- and post-implantation loss:

no effects observed

Total litter losses by resorption:

no effects observed

Description (incidence and severity):

Resorptions were observed in all experimental groups. Intrauterine mortality was comparable in the low and the high dose with the control group and was three times lower in the mid dose than in the control group.

Early or late resorptions:

no effects observed

Dead fetuses:

no effects observed

Description (incidence and severity):

Mortality of 3 fetuses in group 2 (where the resorption number was the lowest) and 1 fetus in group 3 is not high, especially since one of the dead fetuses from group 2 was found to be wrapped around with the umbilical cord and his death cannot be regarded as caused by the test item.

Changes in pregnancy duration:

no effects observed

Changes in number of pregnant:

no effects observed

Other effects:

no effects observed

Description (incidence and severity):

The average weight of the uterus of pregnant females of the treated groups did not differ statistically significantly from average weight of the uterus of pregnant females of the control group.

Details on maternal toxic effects:

The average number of corpora lutea in all treated groups are comparable with the number of corpora lutea in the control group.

Key result

Dose descriptor:

NOAEL

Effect level:

181.8 mg/kg bw/day (actual dose received)

Based on:

test mat.

Basis for effect level:

body weight and weight gain

clinical signs

food consumption and compound intake

mortality

Key result

Abnormalities:

no effects observed

Fetal body weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

Statistically significant increase of weight of fetuses with and without placenta and fetal membranes in treated groups together with the increase of length of fetuses in treated groups (statistically significant in group 2- mid dose) can indicate the test item as inducing the intensive growth of fetuses. This was accompanied by an increase in the number of ossification points in sternum and metacarpus (discussed further). Although an increase of weight in treated versus control group is statistically significant, based on historical and literature data fetuses of all treated groups are within the range of normal body weight of 20 day rat fetuses

Changes in sex ratio:

no effects observed

Changes in litter size and weights:

no effects observed

External malformations:

no effects observed

Skeletal malformations:

no effects observed

Description (incidence and severity):

Variations in sternum ossification centres were of unilateral, bipartite and misaligned character and occurred in all treated groups and in control group. Moreover the incidence of variations in sternum ossification centres in group 1 were comparable with the control group and the occurence of changes was decreasing with the increase of the dose, that allouds to conclude that the test item did not have adverse effect on the process of the ossification of the sternum. Other stated lesions like lack of cranial vault and wavy ribs occurred only in control group without causative effect of the test item.
Analysis of percentage of fetuses with particular number of ossification points of sternum revealed approximately 40% of fetuses with 5 and 6 ossification points in control group and almost 18% of fetuses with 4 ossification points. One can clearly see that in treated groups percentage of fetuses with 6 ossification points increased to over 50% (in group 1 almost 55%, in group 2 the value reached nearly 70% and in group 3 over 60%) and number of fetuses with 5 and 4 ossification points decreased. Statistical analysis of the number of ossification points in sternum confirmed that the average value of ossification points was statistically significantly greater in all treated groups (at low, mid and high dose). This observations lead to the conclusion that the test item influenced on
acceleration of the sternum ossification process.
Analyzing percentage of fetuses with particular number of ossification points in metacarpus, clearly visible is an increase in percentage of fetuses with 4 ossification points and a decrease in percentage of fetuses with 3 ossification points along with the increasing dose of the test item. Statistical analysis of the number of ossification points in metacarpus confirmed that the average value of ossification points was statistically significantly greater in all treated groups and that ossification of metacarpus progressed faster along with increasing doses. Above facts indicate the test item as accelerating ossification of metacarpus of forelimbs. There is no significant difference in ossification of metatarsus of hindlimbs between treated and control groups, because in all groups percentage of fetuses with 4 points of ossification is nearly (control group and group 3) or exactly 100% (group 1 and 2).

Visceral malformations:

no effects observed

Other effects:

effects observed, non-treatment-related

Description (incidence and severity):

The average lengths of the fetuses with tail and lengths of the fetuses without tail were statistically significantly greater in group 2 compared with the control group.

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Basis for effect level:

other: Not adverse effects observed at the highest dose tested.

Key result

Abnormalities:

no effects observed

Key result

Developmental effects observed:

no

Results of mating:

Parameter	Number of females
	Group 0	Group 1	Group 2	Group 3
No. of females	25	25	25	25
No. of pregnants	23	23	23	23
No. of deaths	-	-	-	1
No. of not pregnants	2	2	2	2
Females with resorptions	8	10	4	7

Clinical signs:

Parameter	Number of females
	Group 0	Group 1	Group 2	Group 3
No. of females	25	25	25	25
Coat thinning on back and alopecia on side	-	1	-	-
Coat thinning on forelimbs	-	-	1	-
Coat thinning on abdomen and limbs	-	-	-	1
Respiratory murmurs	-	-	-	12
Distinct decrease of locomotor activity, dejection, bristled coat and difficult respiration	-	-	-	1
Body weight loss – pregnant females	1	-	5	13
Body weight loss – not pregnant females	2	2	2	2

Body weights:

Days of gestation	Number of females
	Group 0	Group 1	Group 2	Group 3
No. of females	23	23	23	23
0	245.22 ± 20.33	247.30 ± 16.02	245.74 ± 13.95	245.65 ± 16.97
5	262.65 ± 23.19	266.83 ± 17.19	265.52 ± 15.44	263.48 ± 17.83
8	271.70 ± 24.06	274.04 ± 17.60	269.70 ± 17.46	260.39 ± 20.55
11	284.52 ± 26.45	289.96 ± 18.61	283.57 ± 17.77	270.22 ± 20.25
14	296.04 ± 26.92	301.65 ± 18.61	295.17 ± 18.79	280.35 ± 18.83*
17	321.35 ± 31.16	329.57 ± 19.43	322.78 ± 20.87	308.35 ± 19.31
20	357.70 ± 37.03	370.83 ± 24.05	359.04 ± 24.66	348.00 ± 20.15

* statistically significant difference with p ≤ 0.05

Food consumption:

Days of gestation	Number of females
	Group 0	Group 1	Group 2	Group 3
No. of females	23	23	23	23
0-5	7.66 ± 0.56	7.81 ± 0.51	7.81 ± 0.56	7.56 ± 0.81
5-8	7.85 ± 0.80	7.87 ± 0.56	7.61 ± 0.55	6.69 ± 1.18*
8-11	7.47 ± 1.00	7.60 ± 0.96	7.21 ± 0.84	6.38 ± 1.13*
11-14	7.92 ± 0.65	7.71 ± 0.69	7.61 ± 0.78	6.56 ± 0.95*
14-17	7.51 ± 0.56	7.33 ± 0.52	7.25 ± 0.36	6.80 ± 0.82*
17-20	6.95 ± 0.57	6.78 ± 0.49	6.73 ± 0.59	6.30 ± 0.71*

* statistically significant difference with p ≤ 0.05

Conclusions:

The NOAEL for maternal toxicity was determined to be 181.8 mg/kg bw/day based on decrease of food consumption and decrease of body weight gain, transient respiratory murmurs and a problable death of one female rat. The test item did not cause adverse effects in fetuses, therefore the NOAEL for fetal toxicity was determined to be 1000 mg/kg bw/day.

Executive summary:

A prenatal developmental toxicity was performed according to the OECD Guideline 414 (GLP study). After a dose range finding, the main test was conducted on 100 females divided into four experimental groups: three treated groups and one control group. Each group consisted of 25 females. From day 5 to 19 of gestation the test item dispersed in distilled water was administered to the females of each group by gavage at 0 (distilled water only), 33.1, 181.8 mg/kg and 1000 mg/kg bw/day. During the study, clinical observations for mortality and signs of toxic influence of the test item were performed in females, their body weight and food consumption were controlled. On day before an expected delivery, on day 20 of gestation, all females were killed with the use of morbital and caesarean section with gross examination. The following endpoints were determined: number of alive and dead fetuses, number of resorptions. After the removing of fetuses each uterus was weighed. The non-gravid uteri and non-gravid horns were overexposed by electric lamp and were stained using ammonium sulphide in order to confirm the non-pregnant status. In the main study the number of corpora lutea was determined in fixed ovaries. The following endpoints were determined in all fetuses: sex, body weight with and without placenta, weight of placenta, body length with and without tail. Each fetus was subjected to gross examination. Half of fetuses from each litter were stained with alizarin and subjected to evaluation of skeleton. The rest of them were evaluated for formation of body cavities and internal organs. Based on the study and analysis of the results it can be stated that the test item did not cause clinical changes and changes in behavior of pregnant females in group 1 and group 2. In group 3 the test item affected respiratory system (what manifested itself by transient respiratory murmurs in twelve females) and probably caused death of one female and clinical symptoms which preceded the death of this female. Additionally, the test item at the highest dose affected decrease of food consumption and decrease of body weight gain in females. Since the statistically significant lower food consumption and lower body weight, clinical signs and death case in group 3 seems to be caused by the test item, the NOAEL for maternal toxicity can be defined as 181.8 mg/kg b.w. Greater average body weights of fetuses in all treated groups and the increase of length of fetuses in group 2 and increases in the average number of ossification points of the sternum and the metacarpus in the treated groups seem to be caused by the test item. However, they are no adverse effects, as they do not diminish an organism’s ability to survive, reproduce or adapt to the environment. Therefore, on the grounds of the used doses, the NOAEL for fetal toxicity can be defined as 1000 mg/kg b.w.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

One key study of Klimisch score 1 is available.

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

Key study:

A prenatal developmental toxicity was performed according to the OECD Guideline 414 (GLP study). From day 5 to 19 of gestation the test item dispersed in distilled water was administered to 25 females per group by gavage at 0 (distilled water only), 33.1, 181.8 mg/kg and 1000 mg/kg bw/day. Based on the study and analysis of the results it can be stated that the test item did not cause clinical changes and changes in behavior of pregnant females in group 1 and group 2. In group 3 the test item affected respiratory system (what manifested itself by transient respiratory murmurs in twelve females) and probably caused death of one female and clinical symptoms which preceded the death of this female. Additionally, the test item at the highest dose affected decrease of food consumption and decrease of body weight gain in females. Since the statistically significant lower food consumption and lower body weight, clinical signs and death case in group 3 seems to be caused by the test item, the NOAEL for maternal toxicity can be defined as 181.8 mg/kg b.w. Greater average body weights of fetuses in all treated groups and the increase of length of fetuses in group 2 and increases in the average number of ossification points of the sternum and the metacarpus in the treated groups seem to be caused by the test item. However, they are no adverse effects, as they do not diminish an organism’s ability to survive, reproduce or adapt to the environment. Therefore, on the grounds of the used doses, the NOAEL for fetal toxicity can be defined as 1000 mg/kg b.w.

Supporting studies:

A combined repeated dose and reproduction/developmental screening was performed on docosanoic acid by the Ministry of Health, Labour and Welfare (Japan, 1998) according to OECD Guideline 422. Rats were orally exposed up to 1000 mg/kg bw/day, males for 42 days and females from 14 days prior to mating to day 3 of lactation. The compound showed no adverse effects on copulation or fertility. No changes related to the dosing of compound were observed in gestation length, gestation index, delivery and lactation. The compound did not demonstrate any adverse effects on the sex ratio, body weights or viability of pups. Also, no morphological abnormalities in pups were observed in any of the treated groups. The NOEL for maternal and reproductive toxicity was determined to be 1000 mg/kg bw/day for both males and females.

A developmental study was performed on rats Scott WJ et al. (1994). Octanoic acid was administered undiluted by oral gavage on day 12 of gestation (single dose). On day 20 of gestation, rats were killed and survivability, number of implantation sites and mean fetal weight were recorded. Octanoic acid was essentially devoid of embryotoxic effects except for a slight reduction of fetal weight. Octanoic acid was non teratogenic under test conditions and the NOAEL was determined to be 18.75 mmol/kg (2700 mg/kg bw).

According to Gottschewski GHM et al. (1967) a vehicle used in coated pharmaceutical tablets was assayed for teratogenicity in rabbits. The vehicle consisted of polyethylene glycol 4000, starch, talcum, silica gel and 5.5% magnesium stearate. Female rabbits received the sample at a dose of 2.5 mg/kg. No significant teratogenic effects were obsrved at the tested dose. The NOAEL for developmental toxicity for magnesium stearate (5.5% in vehicle) was determined to be > 2.5 mg/kg.

In a study performed by Miñana MD et al. (1995) the toxicity effects of prenatal exposure of ammonium acetate was analysed in rats. Female rats were made hyperammonemic by feeding them a diet containing Ammonium acetate (4293 mg ammonium/kg bw/day). Rats were fed the ammonium-containing diet starting on day 1 of pregnancy and were maintained on this diet until weaning (at posnatal day 21). After weaning, pups were either fed a normal diet, with no ammonium acetate added, or continued on ammonium until sacrifice. Rats exposed to NH4+ in utero and during lactation (dams received 4293 mg ammonium/kg/day in the diet from gestational day 1 through lactation day 21), which then received a normal diet, had a statistically reduction in body weight gain by 25 and 16% in male and female offspring, respectively, at 120 days of age. Rats that were continued on the same ammonia diet as their dams had lower body weight gain (25 and 26% for males and females, respectively) at 120 days of age. The NOAEL for developmental toxicity of ammonium acetate in rats was 4293 mg/kg bw/day.

Taking into account the available experimental results, the NOAEL for developmental toxicity for the substance Fatty acids, C16 -18 (even numbered), ammonium salts was determined to be 1000 mg/kg bw/day.

Justification for selection of Effect on developmental toxicity: via oral route:

Only one study available on the test item with Klimisch score of 1.

Justification for classification or non-classification

Based on the available information, the substance is not classified for toxicity to reproduction in accordance with CLP Regulation (EC) no 1272/2008.