Sorbitan stearate

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Remarks:

based on test type (migrated information)

Type of information:

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

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP - Guideline study. According to the ECHA guidance document "Practical guide 6: How to report read-across and categories (March 2010)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Materials and methods

Test guideline

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

Test material

Test material information

Constituent 1

Reference substance name:

Docosanoic acid

EC Number:

204-010-8

EC Name:

Docosanoic acid

Cas Number:

112-85-6

Molecular formula:

C22H44O2

IUPAC Name:

docosanoic acid

Details on test material:

- Name of test material (as cited in study report): docosanoic acid
- Physical state: white powder
- Analytical purity: 85.9 %
- Lot/batch No.: 60805X
- Impurities (identity and concentrations): C14-C20 fatty acids: 10.9 %, C24 fatty acids: 2.3 %
- Storage condition of test material: at room temperature in the dark

Test animals

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

Administration / exposure

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 refrigerated in airtight conditions at each concentration

VEHICLE
- Justification for use and choice of vehicle (if other than water): 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 2 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: 14 days prior to mating until day 3 of lactation

Frequency of treatment:

daily

Details on study schedule:

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

Doses / concentrations

Remarks:

Doses / Concentrations:
100, 300, 1000 mg/kg bw/d
Basis:
actual ingested

No. of animals per sex per dose:

13

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: based on a preliminary 14 day-repeated dose toxicity study, where no signs of toxicity were found

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least once a day

DETAILED CLINICAL OBSERVATIONS: No

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

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

HAEMATOLOGY: Yes (males only)
- Time schedule for collection of blood: prior to autopsy
- Anaesthetic used for blood collection: Yes, pentobarbital sodium
- Animals fasted: 18 - 24 h 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 h 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 and epididymis weight

Litter observations:

STANDARDISATION OF LITTERS
not performed

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: external and internal abnormalities; possible cause of death was determined for pups born or found dead

Postmortem examinations (parental animals):

SACRIFICE
all surviving animals

GROSS PATHOLOGY: Yes
Organ weights:
males: heart, liver, kidneys, thymus, testes, epididymides
females: heart, liver, kidneys, thymus

HISTOPATHOLOGY: Yes (control and 1000 mg/kg bw /day group):
males: heart, liver, spleen, kidney, adrenal, testis, epididymis, brain, thymus, bladder
females: brain, liver, spleen, thymus, kidney, adrenal, bladder, heart, ovary

Postmortem examinations (offspring):

SACRIFICE
- F1 offspring not selected as parental animals was sacrificed at 4 days of age
- these animals were subjected to postmortem examinations (macroscopic examination) including: external and 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

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

100 mg/kg bw/d dose group: significant increase of body weights of males and significant decrease of female body weights during lactation (non-adverse)

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

100 mg/kg bw/d dose group: significant increase of body weights of males and significant decrease of female body weights during lactation (non-adverse)

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

1000 mg/kg bw/d: sporadic findings in brain, heart, liver, spleen, kidney, adrenals, thymus and testes in males and females (non-adverse)

Other effects:

not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Details on results (P0)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
No deaths or abnormalities in general condition were observed in any of the treated groups.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Body weight gain in male animals of the 100 mg/kg bw/day group was higher when compared to the 300 and 1000 mg/kg bw/day dose groups and to the control group (10-33%) over the whole treatment period. Since this effect showed no dose-relation and no other effects were observed in this dose group, it was regarded as non-adverse.
Food consumption of males of the 100 mg/kg bw dose group was about 4-10% increased when compared to controls. A significant decreased food consumption was observed in females of the same dose group during lactation. Since this effect was not dose-related, it was regarded to be not treatment related.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
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 controls.

ORGAN WEIGHTS
A significant liver weight increase (12% (absolute weight), 5% (relative weight)) in males of the 100 mg/kg bw/day dose group and a signficant decrease in kidney weights in females (8% (absolute weight), 3% (non-adervse)) was observed. As there was no dose relationship and no effects were observed at histopathology, they were regarded as non-adverse.

GROSS PATHOLOGY
In males of the 100 mg/kg bw/dose group, diminishment of thymus (1 animal), accessory spleen (1 animal) and liver changes (yellowish colouring (2 animals), pale spot (1 animal), accentuation of lobular pattern (1 animals)) were observed. In males of the 300 mg/kg bw/dose group, enlargment of thyroid (1 animal), pale lung (1 animal), diminishment of thymus (1 animal) and liver changes ((pale area (2 animals), pale spot (1 animal)) were observed. In males of the 1000 mg/kg bw/dose group, diminishment of thymus (1 animal), small and enlarged thymus (1 animal each), pale lung (1 animal) and yellowish colouring of the liver (2 animals) was observed.
Focal constriction of the spleen was observed in two high-dose group females and adrenal enlargement in one female animal of the 100 mg/kg bw dose group. Since the observed effects in male and female animals were found without any dose-relationship and only in few animals, they were regarded as not tretement related.

HISTOPATHOLOGY: NON-NEOPLASTIC
Mycardial degeneration, liver fatty change, changes in spleen and kidney were found to the same extent in few male animals of the highest dose group as compared to males of the control group. One male animal of the highest dose group showed slight fibrosis of the adrenal gland and another one very slight changes of the testes.
The only change differing from controls in females of the high dose group was a thalamus mineralisation in the brain found in one animal. Thus, the effects were regarded as not treatment related.

Effect levels (P0)

Dose descriptor:

NOAEL

Remarks:

fertility

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no substance-related findings were observed

Results: F1 generation

General toxicity (F1)

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

Details on results (F1)

VIABILITY (OFFSPRING)
No change in the number 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 controls.

CLINICAL SIGNS (OFFSPRING)
No clincial signs were observed.

BODY WEIGHT (OFFSPRING)
Body weight of pups of the treatment groups differed not from the controls.

GROSS PATHOLOGY (OFFSPRING)
No morphological abnormalities were found in all groups.

Effect levels (F1)

Dose descriptor:

NOAEL

Remarks:

developmental

Generation:

F1

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no substance-related findings were observed

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

Table 1: Reproductive and offspring indices from dams and pups after oral treatment of parental animals with the test substance in the combined repeated dose and reproductive/developmental screening study (mean ±SD, (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 ratio 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 ratio on day 4	49.8±11.5 (13)	46.3±10.3 (11)	54.8±12.3 (12)	48.9±13.4(13)