Sorbitan C16-18 (even numbered) fatty acid esters, ethoxylated (1-6.5 moles ethoxylated)

Toxicity to reproduction

Administrative data

Endpoint:

one-generation reproductive 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:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study with acceptable restrictions (test substance purity not specified, no skeletal examination of offspring).

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

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

Deviations:

yes

Remarks:

(test substance purity not specified, no skeletal examination of offspring)

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Sorbitan stearate

EC Number:

215-664-9

EC Name:

Sorbitan stearate

Cas Number:

1338-41-6

IUPAC Name:

1,4-anhydro-6-O-stearoyl-D-glucitol

Details on test material:

- Name of test material (as cited in study report): Sorbitan, monooctadecanoate
- Physical state: light yellow crystalline pellet
- Analytical purity: no data
- Storage condition of test material: stored in a sealed box under the room temperature

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 Laboratories Japan, Inc., Yokohama, Japan
- Age at study initiation: 10 weeks
- Weight at study initiation: males (g): 385.3 ± 18.2 - 386.6 ± 16.8, females (g): 220.4 ± 9.6 - 224.8 ± 7.8
- Housing: metal wire mesh cages (220x270x190 mm)
- Diet (ad libitum): CE-2, Clea Japan
- Water (ad libitum): tap water
- Acclimation period: 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.0 - 24.5
- Humidity (%): 55.0 - 65.5
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 / 12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: serial dilutions in water

Details on mating procedure:

- M/F ratio per cage: 1/ 1
- Length of cohabitation: max. 14 days
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Prior to the main study, analytical measurements were performed (GC).

Duration of treatment / exposure:

Females: 2 weeks before mating until day 4 of lactation (~42 days)
males: 42 days

Frequency of treatment:

once daily

Details on study schedule:

- F1 parental animals were not mated.
- males/females: 7 animals were sacrified after treatment and 5 animals were used in a satellite group and sacrified 15 days of recovery period

Doses / concentrations

Remarks:

Doses / Concentrations:
40, 200, 1000 mg/kg bw/d
Basis:
nominal in water

No. of animals per sex per dose:

Males: 0, 7, 40 and 200 mg/kg/d: 12 animals, 1000 mg/kg/d: 7 animals
Males in a satellite group: 0 and 1000 mg/kg: 5 animals
Females: 0, 40, 200 and 1000 mg/kg/d: 12 animals
Females in a satellite group: 0 and 1000 mg/kg/d: 5 animals

Control animals:

yes, concurrent vehicle

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: 1 time daily during the breeding and recovery period and twice daily during the treatment period before and after administration

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule:
males: day 0, 7, 14, 21, 28, 35, 42 during treatment
males and females in satellite group: day 0, 7, 14, 21, 28, 35, 42 during treatment and day 7, 14 during recovery
pregnant females: day 0, 7, 14, 21, 28, 35, 42 during treatment and day 0 and 4 of lactation
[position, posture, spontaneous movement, noise, tremor, ease of retrieval, ease of handling, heart beat, body temperature, fur, skin, visible mucous membranes, tearing, bulging eyes, pupil diameter, postural position, exploratory behavior, grooming, vocalizations, straub tail reaction, walking, stereotypic behavior, bizarre behavior, tremors, piloerection, eye fissure]

BODY WEIGHT: Yes
- Time schedule for examinations:
males: 1, 7, 14, 21, 28, 35, 42 during treatment
males in a sattelite group: 1, 7, 14, 21, 28, 35, 42 during treatment and 1, 7, 14, 15 during recovery
pregnant females: 1, 7, 14, 21, 28 before the copulation, 0, 7, 14, 20 after the copulation and 0, 4, 5 after delivery
females in satellite group: 1, 7, 14, 21, 28, 35, 42 during treatment and 1, 7, 14, 15 during recovery

FOOD CONSUMPTION:
males: between days 1-2, 7-8, 14-15, 29-30, 35-36, 41-42 during treatment
males and females in a satellite group: between days 1-2, 7-8, 14-15, 29-30, 35-36, 41-42 during treatment and 6-7, 13-14 during recovery
females: between days 1-2, 7-8, 14-15 before pregnancy, 0-1, 7-8, 14-15, 20-21 during pregancy and 3-4 during lactation

FOOD EFFICIENCY:
- Body weight gain in g: Yes

WATER CONSUMPTION: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood:
male: one day after the end of dosing period
males and females in satellite groups: 15 day after the end of treatment
females: day 4 during lactation
- Animals fasted: 18 - 24 hours before sacrifice
- How many animals: all animals
- Parameters checked: red blood cell count, white blood cell count, leukocyte classification, amount of hemoglobin, mean corpuscular volume, platelet count, hematocrit, mean corpuscular hemoglobin content, mean corpuscular hemoglobin concentration, prothrombin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood:
male: one day after the end of dosing period
males and females in satellite groups: 15 day after the end of treatment
females: day 4 during lactarion
- Animals fasted: 18 - 24 hours before sacrifice
- How many animals: all animals
- Parameters checked: total protein concentration, total cholesterol concentration, urea nitrogen level, AST (GOT), ALT (GPT), γ-GTP, inorganic phosphorus concentration, sodium ion concentration, potassium ion concentration, chloride ion concentration

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: males: at the end of dosing period, males and females in satellite groups: at the end of recovery period, females: day 4 durgin lactation
- Dose groups that were examined: 0, 40, 200, 1000 mg/kg
- Battery of functions tested: reaction function testing, pupillary reflex, visual orientation, surprised reaction, hind limb retraction reflex, experimental eye (blink) reflex, observed the presence or absence of righting reflex

Oestrous cyclicity (parental animals):

Female: daily observed with vaginal smear specium until mating was confirmed.

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, weight gain, other: general condition,

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 day 42
- Male animasl in a satellite group: All surviving animals day 56
- Maternal animals: All surviving animals day 4 of postpartum
- Femals in a satellite group: All surviving animals day 56

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the thoracic and abdominal viscera.

HISTOPATHOLOGY / ORGAN WEIGHTS
Histopathology of brain, heart, thymus, liver, kidney, spleen tiles, adrenal gland, testis and epididymis were performed and these organ weighed (actual weight) respectively.

Postmortem examinations (offspring):

SACRIFICE
- The F1 offspring was sacrificed on day 4 postpartum.
- These animals were subjected to postmortem examinations.

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the abdominal viscera.

Statistics:

Fisher´s exact test, Mann-Whitney U-grade test, Student´s t-test, Aspin-Welch test, Bartlett test, Dunnett multiple comparison method, Kruskal-Wallis test

Reproductive indices:

estrous cycle, copulation rate, conception rate

Offspring viability indices:

number of pups, offspring viability

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

non-adverse effects

Other effects:

not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Details on results (P0)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
No mortality was observed.
Main groups:
1 male 200 mg/kg/d: loss of fur with ulcer (between day 14 and day 41 during treatment),
Satellite groups:
1 female in the satellite control group: loss of fur with ulcer (between day 14 and day 42), since this change was not found in 1000 mg/kg/d group but in a satellite control group, it was considered as not compound-related.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Main groups:
Females: 200 mg/kg/d: decrease of body weight between day 1 and day 7 during treatment: statistically significant, but was not considered as compound-related change because no abnormality was observed by general condition and food consumption and 1000 mg/kg/d showed no change during treatment phase.
Satellite groups:
Males: 1000 mg/kg/d: increase of body weight between day 7 and day 14 during recovery phase: statistically significant, but was not considered as compound-related change because no change was observed during treatment phase and no abnormality was observed by necropsy.
Females: 1000 mg/kg/d: decrease of body weight between day 7 and day 14 during treatment: statistically significant, but was not considered as compound-related change because no change was observed during treatment phase and no abnormality was observed by necropsy

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)
Oestrous cycle, copulation rate, conception rate: no difference was observed between a control group and other treatment groups.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
40 mg/kg/d: a dam showed abnormality of parturient condition and didn't gather pups and lactate, a second dam showed no abnormality but pups didn't suckle and were dead until day 3 after delivery, 13 pups were born from a third dam but 10 pups didn't suckle and 9 pups were dead until day 2 of lactation, and 4 pups were alive on day 4 of lactation.
But there was no other abnormality of dams in control group, 200 mg/kg/d, and 1000 mg/kg/d groups. Therefore, the effects were considered incidental and not related to treatment.

ORGAN WEIGHTS (PARENTAL ANIMALS)
Males: 200 mg/kg/d decreased relative weight of brain, 1000 mg/kg/d decreased relative weights of epididymis. Otherwise, test substance didn't affect development of brain because no dose-related change was observed, and decreased relative weights of epididymis was not also considered as a compound-related because no change of histopathology test and fertility was observed
at the end of recovery: males: 1000 mg/kg/d increased absolute epididymis weight: this was not also considered as a compound-related because no change of histopathological test and fertility was observed
Females: 1000 mg/kg/d increased absolute brain weight: this was not compound-related because no abnormality of necropsy and histopathological test was observed.

GROSS PATHOLOGY (PARENTAL ANIMALS)
Females: 200 mg/kg/d spots in glandular stomach

HISTOPATHOLOGY (PARENTAL ANIMALS)
Males:
0, 40, 200, 1000 mg/kg/d testes: tubular atrophy and cell debris;
- 1000 mg/kg/d: heart muscle degeneration (to the same extend as in controls), metaplasia in lungs, fat in the periportal hepatocytes (to the same extend as in controls), infiltration of lymphocytes in stomach, extramedullary hematopoiesis and brown pigment in spleen (to the same extend as in controls), basophilic tubules (to the same extend as in controls) and mineral deposition in localized renal cortical (to the same extend as in controls) and circumscribed cyst in kidney, neutrophil and lymphocyte infiltration in prostatic stroma and epithelium (to the same extend as in controls)
- Control group: heart muscle degeneration, neutrophil infiltration in lungs, fat in the periportal hepatocytes, subcapsule necrotic spot in liver, extramedullary hematopoiesis and brown pigment in spleen, basophilic tubules and mineral deposition in localized renal cortical and circumscribed lymphocytic infiltration in kidney, neutrophil and lymphocyte infiltration in prostatic stroma and epithelium
Females:
- 1000 mg/kg/d: myocardial degeneration (to the same extend as in controls), periportal hepatocytes (to the same extend as in controls), extramedullary hematopoiesis (to the same extend as in controls) and brown pigment in spleen (to the same extend as in controls), tubular basophilia (to the same extend as in controls) and circumscribed cyst in kidney
- Control group: myocardial degeneration, periportal hepatocytes, extramedullary hematopoiesis and brown pigment in spleen, tubular basophilia in kidney
These changes were not considered as compound-related.

OTHER FINDINGS (PARENTAL ANIMALS)
Testes, epididymides and ovaries showed no abnormal findings at the end of the study period.

Effect levels (P0)

Dose descriptor:

NOAEL

Remarks:

systemic, fertility

Effect level:

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

Based on:

test mat.

Sex:

male/female

Results: F1 generation

General toxicity (F1)

Clinical signs:

no effects observed

Mortality / viability:

mortality observed, treatment-related

Description (incidence and severity):

but no dose-response effects (non-adverse)

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)
40 mg/kg: 2 dams lost all pups. An additional dam lost 9/13 pups, propably because they did not lactate on day 1. No further mortality was observed, even in the high dose group.

CLINICAL SIGNS (OFFSPRING)
1000 mgkg: one pup with a filamentous tail, this effect was considered as common effect in SD rats and not to be teatement-related.

Effect levels (F1)

Dose descriptor:

NOAEL

Remarks:

systemic

Generation:

F1

Effect level:

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

Based on:

test mat.

Sex:

male/female

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

Table 1: Pup mortality up to day 4 lactation

Dose (mg/kg)	0	40	200	1000
Number of pregnant females	11	11	12	12
Number of pregnant females with live newborns	11	11	12	12
Day 0 of lactation (at birth) Number of newborns	14.2±1.9	13.3±2.4	13.3±2.7	13.3±2.8
Day 4 of lactation Number of live pups	13.8±1.9	10.3±6.1	12.7±3.6	13.3±2.8

Number of live pups in 40 mg/kg showed slightly decreased but it was not statistically significant.

Table 2: Morphological observations of pups

Dose (mg/kg)	0	40	200	1000
Dead pups
Number of dead pups (including missing pups)	4	33	7	0
Number of dead pups with external changes	0	0	0	0
Number of dead pups with visceral changes	0	0	0	0
Live pups
Number of newborns examined (Day 0 of lactation; at birth)	153	134	152	160
No. of newborns with external changes	0	0	0	1
Types and number    Filamentous Tail	0	0	0	1
Number pups examined (Day 4 of lactation; at necropsy)	152	113	152	160
No. of newborns with external changes	0	0	0	1
Types and number   Filamentous Tail	0	0	0	1
No. of pups with visceral changes	0	0	0	0