Glycerides, mixed C8-10 and succinyl

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

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. Neurobehavioural examination was not performed.

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: 10 wks
- Weight at study initiation: 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: NMF, pelleted, ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 8 days

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

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral 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

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

Doses / concentrations

Remarks:

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

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

Observations and examinations performed and frequency:

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

Sacrifice and pathology:

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

HISTOPATHOLOGY: Yes
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.

Statistics:

Barlett test, Dunnett test, Steel test, Fisher test

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

1000 mg/kg bw/day (m): Loosening of the stools was observed.

Mortality:

mortality observed, treatment-related

Description (incidence):

1000 mg/kg bw/day (m): Loosening of the stools was observed.

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

effects observed, treatment-related

Description (incidence and severity):

1000 mg/kg bw/day (f): Increase of blood urea nitrogen was observed; 300 and 1000 mg/kg bw/day (m): On urinalysis, a few males showed high values for protein or were positive for occult blood.

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not examined

Details on results:

CLINICAL SIGNS AND MORTALITY
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 were 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 toxic effects of the test item since these were seen sometimes also in control group.

BODY WEIGHT AND FOOD CONSUMPTION
No changes were detected.

HEMATOLOGY
No changes were detected.

BLOOD CHEMISTRY
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 a toxic 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 a toxic effect of the test item due to lack of dose-dependency. BUN was increased in females of 1000 mg/kg. The test item may have an effect on kidney function.

URIANALYSIS
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. The test item may have an effect on kidney function.

ORGAN WEIGHTS
Absolute weight of adrenals was increased in males of 1000 mg/kg. This change was not regarded as a toxic effect since no histopathological changes were detected and relative weight was not significantly different.
No changes were found in females in the test and control group.

GROSS PATHOLOGY
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. None of these changes was regarded as compound-related.
Adhesion of spleen was observed in one female of control group. Two females and one female showed black patch in stomach at 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 effects.

HISTOPATHOLOGY
Dilatation of seminiferous tubule was observed in one male in 1000 mg/kg. This change was not regarded as a 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 a toxic effect because no change was found in the higher dose groups.
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-dependency was found and these change are observed naturally.

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

100 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: overall effects

2

Dose descriptor:

NOAEL

Effect level:

300 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: overall effects

3

Dose descriptor:

LOAEL

Effect level:

300 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: higher levels of urinary protein

4

Dose descriptor:

LOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: higher levels of blood urea nitrogen

Target system / organ toxicity

Critical effects observed:

not specified