Registration Dossier

Diss Factsheets

Toxicological information

Repeated dose toxicity: oral

Currently viewing:

Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12 March to 16 August 1982
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study conducted in accordance with generally accepted scientific principles, possibly with incomplete reporting or methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1982
Report date:
1982

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: September 1982 pre-Annex V
Deviations:
not specified
GLP compliance:
not specified
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
1-(2,6-bis(4-tolyl)-1,3-dioxano(5,4-d)-1,3-dioxan-4-yl)ethane-1,2-diol
EC Number:
402-950-5
EC Name:
1-(2,6-bis(4-tolyl)-1,3-dioxano(5,4-d)-1,3-dioxan-4-yl)ethane-1,2-diol
Cas Number:
87826-41-3
Molecular formula:
C22 H26 O6
IUPAC Name:
(1R)-1-[(4R,4aR,8aS)-2,6-bis(4-methylphenyl)-hexahydro-[1,3]dioxino[5,4-d][1,3]dioxin-4-yl]ethane-1,2-diol
Details on test material:
- Name of test material (as cited in study report): Gel All-MD.
- Lot/batch No.: 5054006.

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Sankyo Lab Service.
- Age at study initiation: 6 weeks old.
- Weight at study initiation: Average of 185 g for males and 150 g for females.
- Housing: In groups of 3 to 4 and kept in plastic cages (Aircon P from Clea Japan, Inc.) in a breeding room. The floor of the cages were covered with heat sterilised wood shavings (from the Japan Charles River Co), which were changed every 2 - 3 weeks.
- Diet (e.g. ad libitum): MF Rat Feed (Oriental Yeast Company), the test substance was mixed with the diet at the density of 1.25, 2.5 and 5% per weight. Ad libitum.
- Acclimation period: 1 week.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 1°C.
- Humidity (%): 60 ± 10%.
- Air changes (per hr): 12 changes per hour.

IN-LIFE DATES: Day 0 (the day of dosing) up to Day 91.

Administration / exposure

Route of administration:
oral: feed
Vehicle:
other: diet
Details on oral exposure:
(Feeding study)
DIET PREPARATION
- Rate of preparation of diet (frequency): The necessary rat feed for the experiment was prepared all at once, placed in a vinyl bag and stored in a lidded polyethylene tontainer.
- Mixing appropriate amounts with (Type of food): The test substance was mixed with MF Rat Feed (Oriental Yeast Company) at the density of 1.25, 2.5 and 5% per weight.

Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
Not applicable.
Duration of treatment / exposure:
90 days.
Frequency of treatment:
7 days/week.
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
719.8 mg/kg/day (male), 758.8 mg/kg/day (female). Density of substance in food of 1.25%.
Basis:
nominal in diet
Remarks:
Doses / Concentrations:
1518.8 mg/kg/day (male), 1598.2 mg/kg/day (female). Density of substance in food of 2.5%.
Basis:
nominal in diet
Remarks:
Doses / Concentrations:
2925.5 mg/kg/day (male), 3055.9 mg/kg/day (female). Density of substance in food of 5%.
Basis:
nominal in diet
No. of animals per sex per dose:
10 males and 10 females per dose.
Control animals:
yes, plain diet
Details on study design:
Substance Administration:
Unlimited feed, mixed to 1.25%, 2.50% and 5.0% levels of substance and feed with no substance added were given to 4 groups of rats, each with 10 males and 10 females for a period of 90 days. The amount of substance intake was measured once a week by measuring the total feed intake and calculating on an individual basis.
Positive control:
Not applicable.

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS/DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Observed every day, twice a day (morning and afternoon) throughout the study.
- Cage side observations checked: General appearance and behaviour, any signs of abnormalities.

BODY WEIGHT: Yes
- Time schedule for examinations:Once a week.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes.
Determined by the mount of food remaining, adjusted per individual per day.

FOOD EFFICIENCY: No.

WATER CONSUMPTION: Yes
- Time schedule for examinations: Daily.
Determined by the mount of water remaining, adjusted per individual per day.

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At the end of the 90-day period.
- Anaesthetic used for blood collection: Yes (ether).
- Animals fasted: No data.
- How many animals: All the animals.
- Parameters checked: red corpuscle count, platelet count, hematocrit value, hemoglobin, white corpuscle count and the composition of white corpuscles.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At the end of the 90-day period.
- Animals fasted: No data.
- How many animals: All the animals.
- Parameters checked: Glutamic acid-oxalcyl acetic acid-transaminase (GOT), Glutamic acid-pyruvic acid-transaminase (GPT), alkaline phosphatase (Al-p), total protein (TP), blood sugar (Glu), total cholesterol (Cho), nitrogen (BUN), sodium (Na), potassium (K), chlorine (Cl) and calcium (Ca).

URINALYSIS: Yes
- Time schedule for collection of urine: before the start of the administration period, at the 1-month stage, and at the end of the study.
- Metabolism cages used for collection of urine: No data.
- Animals fasted: No data.
- Parameters checked: pH, protein, glucose, ketone bodies, urobilinogen.

NEUROBEHAVIOURAL EXAMINATION: No.

Sacrifice and pathology:
GROSS PATHOLOGY: At the end of the 90-day period, all of the animals were terminated and subject to necropsy.
Organs were weighed immediately after macroscopic examination.


HISTOPATHOLOGY: Yes
The organs were hardended in 10% neutral formalin, covered with parafin, sliced and dyed with hematoxylin-eosin before being examined under an electron microscope. The following were examined:
cerebrum, cerebellum, pituitary gland, thyroid gland, heart, lungs, thymus, liver, kidney, adrenal gland, spleen, testis, prostate gland, bladder, pancreas, ovary, uterus, peritoneum, lymph node, stomach, duodenum, rectum and bone marrow.
Other examinations:
Not reported.
Statistics:
Average values were compared done using t-values.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Mortality:
no mortality 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):
no effects observed
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
no effects observed
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:
no effects observed
Details on results:
COMPOUND INTAKE
Rats consumed a normal amount, demonstrating no aversion to the feed mixed with substance. The average daily intake over the 90 day-period is as below;

Density of analyte Male Female
1.25% 719.8 mg/kg/day 758.8 mg/kg/day
2.50% 1518.8 mg/kg/day 1598.2 mg/kg/day
5.00% 2925.5 mg/kg/day 3055.9 mg/kg/day

For all of the groups the daily substance intake was relatively high at the beginning of the administration period and decreased over time, due to the relationship between weight gain and feed intake. Substance intake for all of the groups decreased by about one-half from the beginning to the end of the administration period as below;

Density of analyte Sex Beginning of period End of period
1.25% Male 1082 mg/kg/day 594 mg/kg/day
1.25% Feale 1003 mg/kg/day 646 mg/kg/day
2.50% Male 2225 mg/kg/day 1160 mg/kg/day
2.50% Female 2104 mg/kg/day 1353 mg/kg/day
5.00% Male 4529 mg/kg/day 2323 mg/kg/day
5.00% Female 4206 mg/kg/day 2509 mg/kg/day

CLINICAL SIGNS AND MORTALITY
There were no deaths in any of the groups. There were no abnormalities observed in any of the groups for behaviour, appearance, coat lustre or excretion.

BODY WEIGHT AND WEIGHT GAIN
All of the female groups had normal rates of growth, with no significant difference relative to the control group.
Among males, the 5.00% group showed an inhibited rate of weight gain around the fifth week, and a statistically significant decrease in the weight gain rate from the eighth week to the end of the experiment. The other groups showed normal weight gain trends throughout, with no difference compard to the control group.

FOOD CONSUMPTION
Daily average feed intake is shown in Table 3, attached. All of the groups consumed normal levels of feed, showing no signs of aversion toward the feed mixed with analyte. In fact, all groups, both male and female, consumed more feed than the control group.

FOOD EFFICIENCY
Not examined.

WATER CONSUMPTION
All of the groups,both male and female, consumed normal levels of water with no significant difference compared to the control group.

OPHTHALMOSCOPIC EXAMINATION
Not examined.

URINALYSIS
The pH, protein and urobilinogen levels of the urine were all within the range of normal values for all groups. There were no differences compared to the readings obtained before substance administration began.
Test for glucose, ketone bodies and occult blood were all negative.

HAEMATOLOGY
All test results fell within the normal range, with no differences observed between the control group and the substance groups.

CLINICAL CHEMISTRY
Statistically significant differences compared to the control group were observed in GOT, alkaline phosphatase (Al-p), total cholesterol (Cho) and calcium (Ca). Statistically significant low GOT values were observed for males in both the 1.25% and 5.00% groups. Statistically significant low Al-p values were observed for males in the 5.00% group; low, though not statistically significant Al-p values for males in the 1.25% and 2.50% groups and females in the 1.25% and 2.50% groups were also observed. However, females of the 5.00% group showed a high Al-p value.
There were no abnormalities in any of the other test items.

NEUROBEHAVIOUR
Not examined.

ORGAN WEIGHTS
None of the organs of the animals were significantly different in weight relative to the control group. However, the carcasses if the 5.00% male group had a statistically significant lowe weight.

GROSS PATHOLOGY
Macroscopic inspection at the time of dissection revealed no abnormalities in any of the rats in both the substance groups and the control groups.

HISTOPATHOLOGY:
Abnormalities were observed in the liver and kidneys.

In the liver, 14 cases of cell infiltration in Glisson's capsule was observed, including in the control group (1/10 in control, 2/10 at 1.25%, 2/10 at 2.5% and 0/10 at 5% in males and 1/10 in control, 3/10 at 1.25%, 1/10 at 2.5% and 4/10 at 5% in females). However, all the cases were extremely minor.

In the kidneys, 16 cases of calcinosis in corticomedullary junction was observed, including in the control group (4/10 in control, 6/10 at 1.25%, 5/10 at 2.5% and 1/10 at 5% in females).

Nephritis was observed in one female in the control group. There were no other abnormalitis observed in any of the other organs.

Effect levels

Dose descriptor:
LOAEL
Effect level:
700 - 750 other: mg/kg/day (daily average consumption in diet)
Sex:
male/female
Basis for effect level:
other: Recommended value by the former Competent Authority: an over cautious LOAEL of 700 - 750 mg/kg could be suggested based on the lowest dose tested. See overall remarks.

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
After unlimited supply of rat feed with Gel ALL-MD content varying from 0.01% to 1.25%, 2.50% , and 5.00%, was fed to rats over a 90-day period, the amount of Gel ALL-MD intake was approximately 750 mg/kg/day for the 1.25% group, 1500 mg/kg/day for the 2.50% group, and 3000 mg/kg/day for the 5.00% group. None of the animals that consumed the analyte demonstrated any toxic effects. The 5.00% male group did show an inhibited weight gain rate after the fifth week and a statistically significant lowering of weight gain rate from the eight week to the end of the 90-day period. As this is within the normal range of values for male SLc-Wister rats, a correlation between inhibited weight gain and dosage level was not established.

No other abnormalities attributable to Gel ALL-MD intake were observed in the urinalyses, blood test, blood serum analysis or examination of the internal organs.


Executive summary:

Three groups of SLc-Wister rats (10 males and 10 females each) were provided unlimited amounts of hard feed containing GEL ALL-MD levels of 1.25%, 2.50% and 5.00% by weight over a 90-day period. A fourth control group was given hard feed with no GEL ALL-MD added.

 The rats consumed normal amounts of feed, demonstrating absolutely no aversion to feed mixed with GEL ALL-MD.

 The daily average consumption of GEL ALL-MD over the 90-day period is shown in the following table.

Density of analyte

Male

Female

1.25%

719.8 mg/kg/day

758.8 mg/kg/day

2.50%

1518.8 mg/kg/day

1598.2 mg/kg/day

5.00%

2925.5 mg/kg/day

3055.9 mg/kg/day

 

There were no cases of death in any of the test groups during the test period, and no abnormal conditions were observed in behaviour, appearance, coat luster or excretion,.

 Males in the 5.00% group showed weight gain rate after the fifth week, and statistically significant decrease in weight gain rate from the eight week to the end of the experiment.

At the end of the 90-day period, microscopic and macroscopic examinations of blood, serum, urine and the internal organs were carried out; no abnormal effects attribute to GEL ALL-MD intake were observed.