Glycerides, mixed C8-10 and succinyl

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: Guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

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:

yes

Remarks:

Neurobehavioural examination was not performed.

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

6106-21-4

Cas Number:

6106-21-4

IUPAC Name:

6106-21-4

Details on test material:

- Name of test material (as cited in study report): Butanedioic acid, disodium salt, hexahydrate
- Physical state: White crystalline
- Analytical purity: 99.9 wt%
- Stability under test conditions: Verified after exposure
- Storage condition of test material: in cool and dark

Test animals

Species:

rat

Strain:

other: CD(SD)IGS

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source:
- Age at study initiation: (P) 10 wks
- Weight at study initiation: (P) Males: 343 - 408 g; Females: 209 - 247 g
- Fasting period before study: No
- Housing: for males and non-preganant females: individual in stainless steel cages
- Diet (e.g. ad libitum): NMF, pelleted, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: at least 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.1 - 25.6 °C
- Humidity (%): 42 - 74%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: injection solvent

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
Dosing solutions were prepared more than once in 6 days. Stability of dosing solution (3 and 200 mg/mL) was verified for 24 hours at 24 °C and for 7 days at 4°C.Test item was diluted in injection water.

VEHICLE
- Lot/batch no. (if required): A005CS

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: max. 4 days
- Proof of pregnancy: vaginal plug and sperm in vaginal smear referred to as day 0 of pregnancy
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged (how): Day 18 of gestation until day 4 of lactation: individual in plastic tray with wood chips

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

At the study initiation, dosing solutions were analyzed and actual concentrations were 91.9 - 98.3% of nominal concentration.

Duration of treatment / exposure:

Male: 14 days before mating, 14 days during mating, 24 days after mating
Females in mating groups: 14 days before mating, 1 - 4 days during mating, 22 or 23 days during resulting pregnancies, 4 days during lactation (total 42 - 46 days)

Frequency of treatment:

daily, 7 days/week

Details on study schedule:

- Age at mating of the mated animals in the study: 12 weeks

Doses / concentrations

Remarks:

Doses / Concentrations:
100, 300 and 1000 mg/kg b.w./day
Basis:
nominal conc.

No. of animals per sex per dose:

6

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Dose levels were based on the results of a foregoing range finding study, in which animals were orally exposed to 0, 30, 100, 300 and 1000 mg/kg bw/day for 14 days (published year unknown, 4802(115-144)). No marked changes were observed even in 1000 mg/kg bw/day group. Therefore, 100, 300 and 1000 were selected as the dose levels for the main study.

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice a day during administration period, and once before necropsy

BODY WEIGHT: Yes
- Time schedule for examinations: males: Day 1, 8, 15, 22, 29, 36, 43, 50 and 53
females: Day 1, 8, 15 before mating, Day 0, 7, 14 and 20 during gestation, Day 0, 4 and 5 during lactation

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

HAEMATOLOGY: Yes
- Time schedule for collection of blood: on day after last administration (Day 53 of males, Day 5 of lactation of females)
- Anaesthetic used for blood collection: Yes (ether)
- Animals fasted: Yes
- How many animals: all administered animals
- Parameters checked: WBC, RBC, Hemoglobin, Hematocrit, MCV, MCH, MCHC, Platelets, Reticulocytes, PT, APTT, Fibrinogen, Differential leukocytes: Lymphocytes, Neutrophils, Eosinophils, Basophils, Monocytes, Large unstained cells

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: on day after last administration or day 15 of recovery period
- Animals fasted: Yes
- How many animals: all administered animals
- Parameters checked: T-potein, Albumin, A/G, Glucose, T-cholesterol, Triglycerides, BUN, Creatinine, T-bilirubin, AST, ALT, ALP, Na, K, Cl, Ca, Inorganic-P, Total bile acid

URINALYSIS: Yes
- Time schedule for collection of urine: In the last week of administration period
- Metabolism cages used for collection of urine: Yes
- Animals fasted: No
- Parameters checked: Volume, Color, pH, Occult blood, Glucose, Protein, Osmotic pressure

Oestrous cyclicity (parental animals):

Estrous cycle length and normality were evaluated in females by vaginal smears prior to mating, and optionally during mating, until evidence of mating was found.

Sperm parameters (parental animals):

Parameters examined in [all/P/F1/F2] male parental generations:
testis weight, epididymis weight, cell type and number per Sertoli cell in seminiferous tubules

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, physical or behavioural abnormalities

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was/was not determined for pups born or found dead.

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals on the day after last administration
- Maternal animals: All surviving animals on Day 5 of lactation

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

HISTOPATHOLOGY / ORGAN WEIGHTS
Histopathology: Skin, heart, lung, trachea, liver, pancreas, esophagus, stomach, duodenum, Payer's patch, colon, small intestine, thymus, spleen, mandibular lymph node, mesenteric lymph node, kidney, urinary bladder, testis, epididymis, ventral prostate, seminal vesicle, piruitary, thyroid, parathyroid, cerebrum, cerebellum, pons, eyeball, sternal bone, femoral bone, sternal bone marrow, femoral bone marrow, spinal cord (sternal, cervical and lumber), mammary gland, sciatic nerve, tongue, ovary, uterus and vagina.
Organ weight: Brain, thymus, liver, spleen, kidneys, adrenals, testes, epididymides, and ovaries.

Postmortem examinations (offspring):

SACRIFICE
- Day 5 of lactation

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

Statistics:

Barlett test, Dunnett test, Steel test, Fisher test

Reproductive indices:

- Copulation index: (number of pairs with successful copulations/number of pairs)x100
- Fertility index: (number of pregnant females/number of pairs with successful copulation)x100
- Implantation index: (number of implantation scars/number of corpora lutea)x100
- Gestation index: (number of dams having live pups/number of pregnant dams)x100
- Delivery index: (number of pups born/number of implantation scars)x100
- Birth index: (number of live pups born/ number of implantation scars)x100

Offspring viability indices:

- Sex ratio at birth: (number of male pups/number of female pups
- Viability index: (number of live pups on Day 4 of lactation/number of live pups born)x100

Results and discussion

Results: P0 (first parental generation)

Details on results (P0)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
No mortality was observed in all groups. Loose stool was observed in one male in 100 mg/kg, four males in 1000 mg/kg and one female in 1000 mg/kg group. Changes in 1000 mg/kg was regarded as test compound related effect. However, the changes in one male in 100 and one female in 1000 mg/kg occurred accidentally. Loss of hair was observed in two males of 300 mg/kg, three males of 1000 mg/kg and several females in all groups including control group. Crust was observed in one male in 300 mg/kg, two males in 1000 mg/kg, one female in 100 mg/kg and one female in 1000 mg/kg. Discharge of eye was observed in one male in 300 mg/kg, one male in 1000 mg/kg. Nasal discharge was observed in two males of 300 mg/kg and two males of 1000 mg/kg. Salivation was observed in one male in 1000 mg/kg and one female in 1000 mg/kg. These changes were not tox effects by the test item since these were seen sometimes also in control group.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
No changes were detected.

HEMATOLOGY (PARENTAL ANIMALS)
No changes were detected.

BLOOD CHEMISTRY (PARENTAL ANIMALS)
Increased sodium was observed in males of 300 and 1000 mg/kg. This change was seemed to be derived by sodium ion of test substance. Therefore, this was not tox effect. Chloride was increased in 300 mg/kg but not marked change. Total bile acid was decreased in males of 1000 mg/kg. This change was caused by quite variable values in the control group. Values in 1000 mg/kg were within scatter ratio of control group. Creatinine was increased in females of 300 mg/kg. This was not tox effect of the test item due to lack of dose-responsibility. BUN was increased in females of 1000 mg/kg. Test item may effect on kidney function.

URIANALYSIS (PARENTAL ANIMALS)
One male showed marked and mild occult blood in 1000 mg/kg and 300 mg/kg, respectively. The relationship between this change and test item was not clear. One male and two males showed higher protein (more than 300 mg/dL) in 300 and 1000 mg/kg, respectively. Test item may effect on kidney function. However, no changes were detected by histopathological findings in kidneys.

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)
One female showed the irregular estrous cycle in 100 and 1000 mg/kg. However, no changes were found in the frequency of irregula cycle occurance.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
No changes were observed between administred groups and control group.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
Duration of gestation was decreased in 100 and 1000 mg/kg. This change was slight and not caused by the test item since all animals showed the normal duration (22 or 23 days).

ORGAN WEIGHTS (PARENTAL ANIMALS)
Absolute weight of adrenals was increased in males of 1000 mg/kg. This change was not regarded as tox effect since no histopahological changes were detected and relative weight was not significantly difference.
No changes were found in females in administered groups and control group.

GROSS PATHOLOGY (PARENTAL ANIMALS)
Diverticulum of small intestine was observed in one male of 1000 mg/kg, red patch/zone of liver in one male of 300 mg/kg, white patch/zone of liver in one male of 1000 mg/kg, enlarged testis in one male of 1000 mg/kg and nodule of epididymis. No changes were regarded as compound-related effect.
Adhesion of spleen was observed in one female of control group. Two females and one female showed black patch in stomach in 300 mg/kg and 1000 mg/kg, respectively. Cyst of pituitary gland was observed in one female of 1000 mg/kg and thin hair in one female of 1000 mg/kg. These changes were also not regarded as compound-related effect.

HISTOPATHOLOGY (PARENTAL ANIMALS)
Dilatation of seminiferous tubule was observed in one male in 1000 mg/kg. This change was not regarded as compound-related effect, since no changes were found in sperm cycle. Atrophy of seminiferous tubule was observed in one male in 300 mg/kg. This change was also not regarded as tox effect because no change was found in higher dosing group.
Necrosis of stomach was observed in females in 300 and 1000 mg/kg. This change was not observed in males. Therefore, this change occurred naturally due to stress of gestation and delivery.
Necrosis of liver and microgranuloma was observed in administered males and females. These changes were not caused by the test item since no dose-responsibility was found and these change are observed naturally.

Effect levels (P0)

1

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 adverse effects on reproduction parameters

2

Dose descriptor:

NOAEL

Remarks:

(paternal)

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: no adverse effects on reproduction parameters

3

Dose descriptor:

NOAEL

Remarks:

(maternal)

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: no adverse effects on reproduction parameters

Results: F1 generation

Details on results (F1)

VIABILITY (OFFSPRING)
No changes were observed between control group and administered groups.

CLINICAL SIGNS (OFFSPRING)
One pup showed anophthalmia and one pup showed polydactyly in 300 mg/kg. However, these incidences were not caused by the test item but natural occurrence.

BODY WEIGHT (OFFSPRING)
Decrease in body weight was observed in males of 100 mg/kg at day 0 after birth, 300 mg/kg at day 0 and 4 after birth, in females at day 4 after birth. However, these changes were regarded as compound related changes since no dose-responsibility was observed.

GROSS PATHOLOGY (OFFSPRING)
Pyelectasis was observed in one dead pup in 100 mg/kg. Red patch of planta was observed in male and female pups 100 mg/kg at day 4 after birth. These pups had the same dam. Therefore, it was not caused by the test item but occurred naturally. Dilated lumen of ureter was observed in male and female pups in administered groups. However, these changes were not caused by the test item but occurred naturally since no dose-responsibility was detected and several litter had the same dam.

Effect levels (F1)

Dose descriptor:

NOAEL

Remarks:

(development)

Generation:

F1

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects on reproduction parameters

Overall reproductive toxicity

Reproductive effects observed:

not specified