Dihydro-3-(tripropenyl)furan-2,5-dione

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

other: Dose range finder

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

2011-06-09 to 2011-07-01

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Well performed study in compliance with the principles of good laboratory practices but not audited by the quality assurance unit

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Deviations:

yes

Remarks:

dose range finder

GLP compliance:

no

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Dihydro-3-(tripropenyl)furan-2,5-dione

EC Number:

295-556-6

EC Name:

Dihydro-3-(tripropenyl)furan-2,5-dione

Cas Number:

92077-08-2

Molecular formula:

R-C4H3O3 , whereas R=C8-C10-alkyl-(branched, unsaturated)

IUPAC Name:

3-[(1E)-2-methyloct-1-en-1-yl]oxolane-2,5-dione

Details on test material:

- Name of test material (as cited in study report): Tripropenyl succinic anhydride
- Molecular weight (if other than submission substance): ca. 220
- Physical state: liquid
- Lot/batch No.: ESD0010823
- Expiration date of the lot/batch: 01.12.2012
- Storage condition of test material: at room temperature
- Other: pH: 1-2; Colour: yellow to brown; Desity 1.01 g/cm3 (20°C, DIN 51757)

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

TEST ANIMALS
- Source: Charles River, 97633 Sulzfeld, Germany
- Age at study initiation: 5-6 weeks
- Weight at study initiation: males: 204-226 g; females: 150-167 g
- Fasting period before study:
- Housing: individually in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding; Full barrier in an air-conditioned room
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Adequate

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C
- Humidity (%): 55 ± 10%
- Air changes (per hr): 10 x
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: To:

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

14 days

Frequency of treatment:

once daily

Doses / concentrations

Remarks:

Doses / Concentrations:
0 (C), 100 (LD), 300 (MD), 600 (HID), 1000 (HD) mg/kg bw
Basis:
actual ingested

No. of animals per sex per dose:

3

Control animals:

yes, concurrent vehicle

Positive control:

no

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least once a day; twice daily all animals were observed for morbidity and mortality except during weekends and on public holidays where observation was made once in the morning.
- Cage side observations checked: The health condition of the animals was recorded.

DETAILED CLINICAL OBSERVATIONS: no data

BODY WEIGHT: Yes
- Time schedule for examinations: once before assignment to the experimental groups and on study days 1, 7 and 14 during the treatment period as well as on the day of necropsy.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Food consumption was measured on study days 1, 7 and 14.
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No
- Time schedule for examinations:

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at the terminal sacrifice of all surviving animals
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes
- How many animals: all surviving animals
- Parameters checked: haematocrit value (HCT), haemoglobin content (Hb), red blood cell count (RBC), platelet count (PLT), white blood cells (WBC)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: On day of terminal sacrifice
- Animals fasted: Yes
- How many animals: all surviving animals
- Parameters checked: alanine aminotransferase (ALAT), aspartate-aminotransferase (ASAT), alkaline phosphatase (AP), creatinine (Crea), total protein (TP), albumin (Alb), urea, total cholesterol (Chol), sodium (Na), potassium (K)

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER:

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
On study day 15, all surviving animals of the study were subjected to a detailed gross necropsy which included careful examination of the external surface of the body, all orifices and the cranial, thoracic and abdominal cavities and their contents. All macroscopic findings were recorded and organs showing gross abnormalities were preserved in 10 % neutral buffered formalin for a possible histopathological evaluation.
All animals found moribund and intercurrently sacrificed were also subjected to a gross necropsy.
The wet weight of the following organs was taken from all terminally sacrificed animals as soon as possible. Paired organs were weighed separately. Organ weights of animals found dead or euthanised for ethical reasons were taken.
Organ weighed at necropsy:
liver, kidneys, adrenals, testes, epididymides, prostate and seminal vesicles with coagulating gland (as a whole), thyroid/parathyroid glands, ovaries, uterus with cervix (females), thymus, spleen, heart, brain.

HISTOPATHOLOGY: no

Statistics:

In the evaluation of laboratory parameters, all values within a range of the mean value ± the two fold standard deviation (x ± 2s) are considered to be „normal“ values within a „normal“ population.

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Mortality:

mortality observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Details on results:

CLINICAL SIGNS AND MORTALITY
Three male from HD and 5 female animals from HID and HD group died during the 14 days treatment period. one female animal HID group was sacrificed due to animal welfare reasons on day 4.
Majority of male and female animals from various treated groups showed test item related clinical signs. Predominant clinical signs observed were reduced spontaneous activity, piloerection, salivation, moving the bedding and nasal discharge.

BODY WEIGHT AND WEIGHT GAIN
In male and females, slight decrease in group mean overall body weight change (Day 1-14) was observed in HID and MD group respectively when compared with corresponding controls.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
In HID males and LD, MD females, slight decrease in food consumption was observed during the both weeks of the treatment when compared with the controls.

HAEMATOLOGY
White Blood Count (WBC):
All mean and individual values of male and females (except individual value for one female from LD group) were within the biological range (3.3-21.5 x 10³/μL). Female WBC values from LD and MD were comparable with the controls. However, dose dependent decrease in male WBC values was observed when compared with controls.
Platelet:
All mean values and individual values of male and females were within the biological range (370-1856 x 10³/μL). However, a slight dose-dependent increase in platelet count was observed in MD and HID group of male and LD and MD group of female when compared to the controls. This effect was also correlated with reduction in spleen weights in these groups and therefore could be considered as a treatment related effect.
All group mean and individual values of other parameters were comparable and within the biological range

CLINICAL CHEMISTRY
ASAT:
All mean and individual ASAT/SGOT values of male and female groups were within the biological range (18-215.3 U/L). However, slightly decrease was observed in female LD and MD groups when compared with controls.
ALAT:
All mean and individual ALAT/SGPT values of male and female groups were within the biological range (8-102 U/L). However, slightly decrease was observed in female LD and MD groups when compared with controls.
AP:
All mean and individual values of male and female groups were within the biological range (51- 769.8 U/L). However, slight decrease was observed in male HID and female MD groups when compared with controls.
UREA:
All mean and individual UREA values of male and female groups were within the biological range (3.18-31.4 mmol/L). However, dose dependent increase was observed in female LD and MD groups when compared with controls.
CREA:
All mean and individual CREA values of male and female (except female No. 19 from LD) groups were within the biological range (12-80 μmol/L). However, dose dependent increase was observed in female LD and MD groups when compared with controls
CHOL:
Majority of mean values and individual CHOL values in male (except mean values from C, MD and HID) and females were below the biological range (1.02-14.6 mmol/L).
All mean and individual values of other parameter determined in all groups were comparable and within the biological range.

ORGAN WEIGHTS
Spleen:
Absolute and relative spleen weights in male and females of treated groups were slightly decreased when compared with controls.
Kidney:
No biologically significant difference was observed in absolute and relative kidney weights in males of treated groups when compared with controls. However, slightly increase in absolute and relative kidney weights were observed in MD females when compared with controls.
Epididymides:
Slightly decrease in absolute and relative epididymides weights were observed in males of treated groups when compared with controls.
Uterus with cervix:
Absolute and relative weights in females of MD group were slightly decreased when compared with controls.
Prostate, seminal vesicles with coagulating gland (as a whole):
Absolute and relative prostate, seminal vesicles with coagulating gland weights in male of treated groups were slightly decreased when compared with controls
Heart:
Absolute and relative heart weights in male and females of treated groups were slightly decreased when compared with controls.
No effects were obeserved in other ograns.

GROSS PATHOLOGY
Few specific gross pathological changes were recorded for the male HD and female HID and HD animals. Predominant findings observed were reddish discoloration, swelling, gas and liquid filled digestive tract.

Effect levels

Dose descriptor:

LOAEL

Effect level:

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

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: overall effects mortality; body weight; organ weights

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

In conclusion, the repeated dose administration of Tripropenyl succinic anhydride to male and female Wistar rats at dosages of 100, 300, 600 and 1000 mg/kg body weight for 14 days revealed toxicological findings, clinical signs and mortalities in both male and females.

Executive summary:

In this 14-day dose range finding study, the test item was administered daily by oral gavage to Wistar rats of both sexes at dose levels of 100 (LD), 300 (MD), 600 (HID), 1000 (HD) mg/kg bw. One group receiving the vehicle corn oil served as control (C). The animals were observed for signs of toxicity, body weight and food consumption during the study. All surviving animals were sacrificed on day 15 and were subjected to a detailed gross necropsy. All found dead animals, moribund and intercurrently sacrificed animals were also subjected to a gross necropsy. The wet weight of the organs was taken from all terminally sacrificed animals as soon as possible. Haematological and clinical biochemistry examinations were made on blood samples obtained from the overnight fasted surviving animals at the terminal sacrifice.

The treatment revealed toxicological findings, clinical signs, and mortalities in both male and females ( 3 male in HD group and 6 females in HID and HD groups). Other predominant toxicologcial findings observed were:

Decrease in overall body weight change and food consumption in HID males and LD, MD females. Dose dependent decrease in male WBC values, a slight dose dependent increase in platelet count in MD and HID group of male and LD and MD group of female. Slightly decrease in ASAT, ALAT and increase in urea, creatanine in female LD and MD. Slight decrease in AP in Male HID and female MD. Slight decrease in absolute and relative spleen and heart weights in male and females of treated groups. Slightly increase in absolute and relative kidney weights in MD females. Slightly decrease in absolute and relative epididymides and prostate, seminal vesicles with coagulating gland weights in males of treated groups.

This study provided a basis for the selection of dose levels for 28 -day repeated doase oral toxicity study and/or reproductive/developmental toxicity screening test.