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Endpoint summary

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Effects on fertility

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

For the read-across justification see section 13.

Reason / purpose for cross-reference:

read-across source

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:

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 per 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:

no effects observed

Mortality:

no mortality observed

Description (incidence):

- 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 weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

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.

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Reproductive performance:

no effects observed

Description (incidence and severity):

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.

Key result

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 treatment-related effects were observed

Mortality / viability:

no mortality observed

Description (incidence and severity):

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 and weight changes:

no effects observed

Description (incidence and severity):

No difference in bodyweight of pups of the dose groups and the control group was found.

Gross pathological findings:

no effects observed

Description (incidence and severity):

No morphological abnormalities were found in all groups.

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

>= 1 000 mg/kg bw/day (nominal)

Based on:

test mat. (total fraction)

Sex:

male/female

Basis for effect level:

other: no treatment-related effects were observed

Key result

Reproductive effects observed:

no

Lowest effective dose / conc.:

1 000 mg/kg bw/day (nominal)

Treatment related:

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)

Conclusions:

No treatment-related effects were observed. Thus the NOAEL can be stated to be >= 1000 mg/kg bw nominal.
Thus the NOEL value for Reaction mass of stearic acid and stearic anhydride is comparable.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effects on developmental toxicity

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available