Registration Dossier

Toxicological information

Toxicity to reproduction

Currently viewing:

Administrative data

Endpoint:
one-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions Analytical purity of test substance not reported
Cross-reference
Reason / purpose:
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:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
yes
Remarks:
analytical purity of test substance not reported
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
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 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 (7 a.m. to 7 p.m.)

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: serial dilutions in water were prepared
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: 7 animals were sacrified after treatment
males in a satellite group: 5 animals were sacrified 15 days of recovery period
females: 12 animals are sacrified after treatment
females in a satellite group: sacrified 15 days of recovery period
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 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 a 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 anatomy
- 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, Rotoronpin 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 anatomy
- 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 animals)

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
Gross pathological findings:
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
1 male 200 mg/kg/d: loss of fur with ulcer (between day 14 and day 41 during treatment), 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 found in a satellite control group, it was not compound-related change.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Males satellite: 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: 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.
Females satellite: 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 (FB02005) showed abnormality of parturient condition and didn't gather pups and lactate, another dam (FB02007) showed no abnormality but pups didn't suckle and were dead until day 3 after delivery, 13 pups were born from another dam (FB02012) 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, test substance did not effect on delivery and lactation.

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 (same as control), metaplasia in lungs, fat in the periportal hepatocytes (same as control), infiltration of lymphocytes in stomach, extramedullary hematopoiesis and brown pigment in spleen (same as control), basophilic tubules n (same as control) and mineral deposition in localized renal cortical (same as control) and circumscribed cyst in kidney, neutrophil and lymphocyte infiltration in prostatic stroma and epithelium (same as control)
- Control group: heart muscle degeneration, neutrophil infiltration in lugs, 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 (same as control), periportal hepatocytes (same as control), extramedullary hematopoiesis (same as control) and brown pigment in spleen (same as control), tubular basophilia (same as control) 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
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):
no dose-response effects
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: all pups from two dams (FB02005, FB02007) were dead, all pups from FB02012 didn't suckle on day 1 of lactation and 9 pups were dead, but 4 pups were alive until day 4 of lactation. No deaths in the higher dose groups.

CLINICAL SIGNS (OFFSPRING)
1000 mgkg: one pup with a filamentous tail
It happens naturally in this species, therefore, it was not caused by the test material

Effect levels (F1)

Dose descriptor:
NOAEL
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

Number of pups 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.

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

Applicant's summary and conclusion