Bis-O-(benzylidene)-D-glucitol

Endpoint summary

Description of key information

The following test data has been used to assess the repeated dose toxicity of the substance.

- reading across the results of a 28 -day repeated dose toxicity study conducted on a structurally similar analogue substance.

- 3-month subactue toxicity study of substance itself by oral administration to mice and rats.

28 -day repeated dose toxicity study:

Source Substance; Gel All DX (EC-413 -110 -2):

A 28-day repeated-dose oral toxicity study of the test material followed by a 14-day recovery test was conducted in male and female Crj: CD (SD) rats (6/sex/ group), 5 weeks of age at the start of dosing. The highest dose was set at 1000 mg/kg, and 3 lower doses at 200, 40 and 8 mg/kg. Recovery groups were separately provided for vehicle control and 1000 mg/kg groups.

There were no deaths on account of administration of the test material.

No abnormalities were noted in general conditions, body weights, food consumption, haematological examinations, blood chemical examinations, urinalysis, necropsy and histopathological examinations.

In organ weights, increased absolute and relative liver weights were noted in males of the 1000 mg/kg group.

In the recovery test, no abnormalities were noted.

In conclusion, No Observed Effect Level of the test material for rats was considered to be 200 mg/kg/day under the conditions tested.

The No Observed Adverse Effect Level was considered to be 1000 mg/kg/day.

3-month subactue toxicity study by oral administration to mice and rats:

A 3-month subacute (oral feed) study of dibenzyliden sorbitol was conducted in mice and rats, and the following results were obtained.

No special changes were noted in the general symptoms, food ingestion volume, water-intake volume, body weight change, urinalysis findings, etc. The blood and serum biochemistry tests indicated significant differences in some parameters both in mice and rats but all these changes were within the physiological range of variation and indicated no relation to the administration of D.B.S. Macroscopic autopsy findings detected some lesions in the kidney, liver, lung and bladder, etc. of some animals but there was no change specifically observed in the dose groups. As to the organ weights, the weight of liver tended to increase slightly but there was no other tendency that was commonly observed in the males and females of mice and rats and that demonstrated a proportionally certain relation to the dose in the remaining organs.

As to the histopathological findings, some localized abnormal findings were obtained from the lung, kidney, heart, liver, pancreas and adrenal gland but all of these events occurred as single episodes and there was no specific change to indicate the correlation to the administration of D.B.S.

As described in the above, the results of a 3-month subacute toxicity study by oral administration of D.B.S. indicated that dibenzyliden sorbitol was very stable and had hardly any toxicity. The maximum dose that does not demonstrate the toxicity in such case is 3,200 mg/kg/day for the male and female mice, and 2,000 mg/kg/day for the male and female rats.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

13 January 1995 to 18 July 1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP guideline study.

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

other: 28-day Repeated Dose Toxicity Study in Mammalian Species of The Notification on Partial Revision of Testing Methods Relating to New Chemical Substances (Notification No. 700 of Kanpogyo, No. 1039 of Yakuhatsu, and No. 1014 of of 61 Kikyoku, Dec. 5, 1986)

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Japan, Inc.
- Age at study initiation: 5 week
- Weight at study initiation: males: 144.4- 177.4 g; females: 116.4- 137.7 g
- Housing: animals were housed individually in hanging stainless steel cages with wire-mesh floor
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: animals were quaratined and acclimatised

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 2 °C
- Humidity (%): 55 ± 10 %
- Air changes (per hr): 10- 15
- Photoperiod (hrs dark / hrs light): 12/ 12

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: The test substance was weighted accurately and mixed with olive oil to make a concentration of 10.0 w/v %. This was prepared once a week. Other three concentrations of 2.0, 0.4 and 0.08 w/v % were prepared every time from 10.0 w/v %.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The stability and uniformity of the test substance in these preparations was confirmed in the laboroatory.

Duration of treatment / exposure:

28 days

Frequency of treatment:

Once daily for 28 day, and the subsequent 14 days were used as a recovery period

Dose / conc.:

8 mg/kg bw/day (nominal)

Remarks:

Low dose

Dose / conc.:

40 mg/kg bw/day (nominal)

Remarks:

Intermediate dose (I)

Dose / conc.:

200 mg/kg bw/day (nominal)

Remarks:

Intermediate dose (2)

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

High dose

No. of animals per sex per dose:

6 males and 6 females per dose.

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: A 14-day repeated-dose preliminary toxicity study was carried out at 3 doses of 50, 250 and 1000 mg/kg. There were abnormalities in organ weights in the 250 mg/kg and higher groups, and blood chemical examinations in the 50 mg/kg and higher groups.
In the main study, the maximum dose was chosen at 1000 mg/kg and 3 lower doses at 200, 40 and 8 mg/kg. Recovery groups were set at 1000 mg/kg and the vehicle control groups.

Positive control:

None.

Observations and examinations performed and frequency:

The day of the start of dosing was defined as day 1, and the day before as day -1. The week of the start of dosing period was defined as week 1. Also, the next day of final dosing was defined as recovery day 1, and the week of the start of recovery period as recovery week 1.

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least once per day

BODY WEIGHT: Yes
- Time schedule for examinations
- Before dosing: day -2 (at the time of grouping)
- During the dosing period: days 1 (at the start of dosing), 3, 5, 8, 10, 12, 15, 17, 19, 22, 24, 26 and 28
- During the recovery period: days 1 (at the start of the recovery period), 3, 5, 8, 10, 12 and 14
- Immediately before necropsy for calculation of relative organ weights

FOOD CONSUMPTION:
- Time schedule for measurement of food consumption
- Before dosing: Once
- During the dosing and recovery periods: Twice a week

HAEMATOLOGY: Yes
- Anaesthetic used for blood collection: Yes
- Animals fasted: Yes, overnight (16- 20 h) at the end of the dosing and recovery periods
- How many animals: All
- Blood samples were taken via abdominal aorta from rats under ether anesthesia. Sodium citrate was used as an anticoagulant for examinations of prothrombin time and activated partial thromboplastin time, and EDTA-2K was used for another parameters.
- The following parameters were examined: red blood cell count (RBC), white blood cell count (WBC), haemoglobin conc. (Hb), haematocrit value (Ht), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), platelet count, reticulocytes count, prothrombin time (PT), activated partial thromboplastin time (APTT), differentiation of leukocytes (band form neutrophils (N-Band), segmental neutrophils (N-Seg), eosinophils (Eosino), basophils (Baso), lymphocytes (Lymph), monocytes (Mono)).

CLINICAL CHEMISTRY: Yes
Sera were separated from the blood samples used in hematological examinations and examined as follows.
- Parameters checked: GOT, GPT, alkaline phosphatase (ALP), cholinesterase (ChE), γ-GTP, total cholesterol (T-Cho), triglyceride (TG), glucose, total protein (T-Ptotein), albumin, A/G ratio, blood urea nitrogen (BUN), creatinine, total bilirubin (T-Bil), Ca, IP, Na, K, Cl

URINALYSIS: Yes
- Time schedule for collection of urine: 16-h urine samples were collected from all animals at day 28 and recovery day 14
- Metabolism cages used for collection of urine: Yes
- Urine was examined for: volume, colour, additional items of pH, protein, ketone bodies, bilirubin, occult blood, glucose and urobilinogen

ORGAN WEIGHTS
- The following organs were weighed wet in all animals: brain, liver, spleen, kidneys, adrenal glands, testes (or ovaries)

HISTOPATHOLOGICAL EXAMINATIONS
- The following organs and tissues from all animals were preserved in 10 % formalin: brain (cerebrum, cerebellum), hypophysis, eyeball, thyroid glands (with parathyroid glands), heart, lung, liver, kidneys, spleen, adrenal glands, stomach, intestine (duodenum to rectum), testes (or ovaries), urinary bladder, bone marrow (femur), gross lesions
- Light microscopic examinations were performed on the following organs and tissues after paraffin embedding and sectioning followed by hematoxylin and eosin staining:
- Dosing period
- Vehicle control and 1000 mg/kg groups: liver, spleen, kidneys, heart, stomach, intestine (duodenum, jejunum, ileum, cecum, colon, rectum), adrenal glands
- Gross lesions
- Dosing period, male 8 mg/kg group: glandular stomach; male 40 mg/kg group: cerebrum; male 200 mg/kg group: skin; female 8 mg/kg group: glandular stomach
- Recovery period, male 1000 mg/kg group: glandular stomach

Sacrifice and pathology:

NECOROSPY:
All animals were necropsied in detail to record.

ORGAN WEIGHTS
- The following organs were weighed wet in all animals: brain, liver, spleen, kidneys, adrenal glands, testes (or ovaries)

HISTOPATHOLOGICAL EXAMINATIONS
- The following organs and tissues from all animals were preserved in 10 % formalin:
brain (cerebrum, cerebellum), hypophysis, eyeball, thyroid glands (with parathyroid glands), heart, lung, liver, kidneys, spleen, adrenal glands, stomach, intestine (duodenum to rectum), testes (or ovaries), urinary bladder, bone marrow (femur), gross lesions

- Light microscopic examinations were performed on the following organs and tissues after paraffin embedding and sectioning followed by hematoxylin and eosin staining:
Dosing period:
- Vehicle control and 1000 mg/kg groups: liver, spleen, kidneys, heart, stomach, intestine (duodenum, jejunum, ileum, cecum, colon, rectum), adrenal glands
Gross lesions:
- Dosing period, male 8 mg/kg group: glandular stomach; male 40 mg/kg group: cerebrum; male 200 mg/kg group: skin; female 8 mg/kg group: glandular stomach
- Recovery period, male 1000 mg/kg group: glandular stomach

Statistics:

Data regarding body weights, food consumption, hematological examination, blood chemical examination, urine volume and organ weights were analyzed using Bartlett's test for homogeneity of variance. If the variances were homogeneous at a significance level of 5 %, one way analysis of variance was performed. When there was a significant difference in this analysis, the difference between the vehicle control group and each of the treatment group was analyzed by Dunnett's test (equal number of data) or Scheffe's test (unequal number of data).

If the variances were not homogeneous in the Bartlett's test, Kruskal-Wallis's test was used. When there was a significant difference in this test, the difference between the vehicle control group and each of the treatment group was analyzed by nonparametric Dunnett's test (equal number of data) or nonparametric Scheffe's test (unequal number of data).

Clinical signs:

no effects observed

Description (incidence and severity):

No toxicologically significant effects

Mortality:

no mortality observed

Description (incidence):

No toxicologically significant effects

Body weight and weight changes:

no effects observed

Description (incidence and severity):

No toxicologically significant effects

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

Description (incidence and severity):

No toxicologically significant effects

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

No toxicologically significant effects

Urinalysis findings:

no effects observed

Description (incidence and severity):

No toxicologically significant effects

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Increased relative liver weight in males in the 1000 mg/kg group.

Gross pathological findings:

no effects observed

Description (incidence and severity):

No toxicologically significant effects

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

No toxicologically significant effects

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
There were no deaths.

During the dosing period:
- Male: Salivation immediately after dosing was noted in the vehicle control group (3/12, days 12, 24 and 25) and the 1000 mg/kg group (2/12, days 25 and 28). Scab formation (2/6), loss of hair (2/6) and exudate (1/6) were also noted in the 200 mg/kg group.
- Female: Salivation was noted in the vehicle control (5/12, between day 13 and 27) , the 40 mg/kg group (3/6, between day 12 and 26) and the 1000 mg/kg group (1/12, days 27 and 28). These conditions appeared immediately after dosing.

During the recovery period;
- Male: Dark reddish change of right eyeball (1/6) was noted in the vehicle control group.
- Female: Loss of hair (1/6) was noted in the 1000 mg/kg group.

BODY WEIGHT AND WEIGHT GAIN
During the dosing period
- No abnormalities were noted in either sex of any dosing group.

During the recovery period
- Male: No abnormalities were noted in any dosing group.
- Female: A significant increase of body weight gain on day 3 was noted in the 1000 mg/kg group.

FOOD CONSUMPTION
During the dosing period
- No abnormalities were noted in either sex of any dosing group.

During the recovery period
- Male: No abnormalities were noted in any dosing group.
- Female: A significant increase of food consumption gain on day 4 was noted in the 1000 mg/kg group.

HAEMATOLOGY
- The blood sample of 1 female was not used in a statistical analysis because of a part of coagulation.

At the end of the dosing period
- No abnormalities were noted in either sex of any dosing group.

At the end of the recovery period
- Male: No abnormalities were noted in any dosing group.
- Female: A decrease in mean corpuscular haemoglobin concentration was noted in the 1000 mg/kg group

CLINICAL CHEMISTRY
At the end of the dosing period
- Male: No abnormalities were noted in any dosing group.
- Female: An increase in K was noted in the 40 and 200 mg/kg groups

At the end of the recovery period
- Male: No abnormalities were noted in any dosing group.
- Female: Decreases in Cholinesterase, albumin and A/G ratio and increases in γ-GTP and creatinine were noted in the 1000 mg/kg group.

URINALYSIS
- There were no change considered to be due to the test substance during the dosing period and at the end of the recovery period.

ORGAN WEIGHTS
At the end of the dosing period
- Male: An increase of absolute liver weight was noted in the 40 mg/kg or higher groups and an increase of relative liver weight was noted in the 1000 mg/kg group.
- Female: No abnormalities were noted in any dosing group.

At the end of the recovery period
- Male: No abnormalities were noted in any dosing group.
- Female: An increase of absolute liver weight and decreases of relative kidney and brain weights were noted in the 1000 mg/kg group.

GROSS PATHOLOGY (NECROSPY)
At the end of the dosing period
- Male: Blackish spots of mucosa in the glandular stomach (1/6) in the 8 mg/kg group, dilatation of ventricules in the cerebrum (1/6) in the 40 mg/kg group and scab formation (2/6) in the 200 mg/kg group were noted.
- Female: Blackish spot of mucosa in the glandular stomach (1/6) was noted in the 8 mg/kg group.

At the end of the recovery period
- Male: Blackish spot of mucosa in the glandular stomach (1/6) was noted in the 1000 mg/kg group.
- Female: No abnormalities were noted in any dosing group.

HISTOPATHOLOGY
At the end of the dosing period
- Male: Cyst formation in the kidney in the vehicle control group (2/6) and the 1000 mg/kg group (1/6), necrosis of mucosa in the glandular stomach in the 8 mg/kg group, dilatation of ventricles in the cerebrum (1/1) in the 40 mg/kg group and necrosis and scab formation in the skin (2/2) in the 200 mg/kg group.
- Female: Cyst formation in the kidney in the vehicle control group (2/6) and the 1000 mg/kg group (1/6) and necrosis of mucosa in the glandular stomach in the 8 mg/kg group were noted.

At the end of the recovery period
- Necrosis of mucosa in the glandular stomach was noted in males of the 1000 mg/kg group. Females were not examined.

Dose descriptor:

NOEL

Effect level:

200 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Increased relative liver weight in males of the 1000 mg/kg group

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Increase in relative liver weight in males at 1000 mg/kg is not considered to be a serious adverse effect as there was no associated organ dysfunction and no elevated liver weight in 1000 mg/kg males at the end of the recovery period.

Key result

Critical effects observed:

no

Conclusions:

The No Observed Effect Level (NOEL) of the test material for rats was determined to be 200 mg/kg/day under the conditions tested.

Executive summary:

A 28-day repeated-dose oral toxicity study of the test material followed by a 14-day recovery test was conducted in male and female Crj: CD (SD) rats (6/sex/ group), 5 weeks of age at the start of dosing. The highest dose was set at 1000 mg/kg, and 3 lower doses at 200, 40 and 8 mg/kg. Recovery groups were separately provided for vehicle control and 1000 mg/kg groups.

There were no deaths on account of administration of the test material.

No abnormalities were noted in general conditions, body weights, food consumption, haematological examinations, blood chemical examinations, urinalysis, necropsy and histopathological examinations.

In organ weights, increased absolute and relative liver weights were noted in males of the 1000 mg/kg group.

In the recovery test, no abnormalities were noted.

In conclusion, No Observed Effect Level of the test material for rats was considered to be 200 mg/kg/day under the conditions tested.

The No Observed Adverse Effect Level was considered to be 1000 mg/kg/day.

Reference 2

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

Please see Section 13 for "Read-across justification to support the REACH registration of Bis-O-(benzylidene)-D-glucitol (EC 251-136-4) at 10-100 tpa" for full details.

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
It is proposed that the structural similarity and properties of the target substance and the structural analogue (sources substance) are sufficiently close for there to be a reasonable expectation of similar effects.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Source substance:
IUPAC name: 1,3:2,4-bis-O-((3,4-dimethylphenyl)methylene) D-Glucitol
EC number: 413-110-2
CAS number: 135861-56-2


Target Substance: Bis-O-(benzylidene)-D-glucitol
IUPAC name: 1,3:2,4-bis-O-dibenzylidene-D-glucitol
EC number: 251-136-4
CAS number: 32647-67-9


3. ANALOGUE APPROACH JUSTIFICATION
Based on the structural similarity of the source substances and target substance, similarity of physic-chemical properties and similarity in experimental (eco)toxicological test data (and toxicokinetic behaviour assessment) it is concluded that target substance and the structural analogue (source substance) are sufficiently close for there to be a reasonable expectation of similar effects, for the endpoints where results have been read-across.

4. DATA MATRIX
Please see Section 13 for "Read-across justification to support the REACH registration of Bis-O-(benzylidene)-D-glucitol (EC 251-136-4) at 10-100 tpa" for full details.

Reason / purpose for cross-reference:

read-across source

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):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Dose descriptor:

NOEL

Effect level:

200 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Increased relative liver weight in males of the 1000 mg/kg group

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Increase in relative liver weight in males at 1000 mg/kg is not considered to be a serious adverse effect as there was no associated organ dysfunction and no elevated liver weight in 1000 mg/kg males at the end of the recovery period.

Key result

Critical effects observed:

no

Discussion

The oral toxicity of the test material (structural analogue source substance EC 413-110-2)

was examined in SD strain male and female rats given daily doses of 8, 40, 200 and 1000 mg/kg for 28 d, followed by the 14-d recovery period.

There were no deaths on account of administration of the test material.

No abnormalities were noted in body weights and food consumption during the dosing period, and hematological examinations, blood chemical examinations, urinalysis and necropsy at the end of the dosing period.

In general conditions, salivation, which noted in both sexes of the 1000 mg/kg group, had slightly toxic effect since it was noted just after dosing and in vehicle control group.

In organ weights, as the dose-related changes, increased absolute and relative liver weights were noted in males of the 1000 mg/kg group. These were considered as the very slightly effect of the test substance since no other related changes were noted. An increase of absolute liver weight in males of the 40 and 200 mg/kg groups were not considered to be effect of the test substance for no changes in relative liver weight.

In the histopathological examinations, cyst formation in the kidney was noted in both sexes of the 1000 mg/kg group at end of the dosing period. But it was not considered as the effect of the test substance because this change was common in this strain and noted in the vehicle control group.

In the recovery test, decreases in mean corpuscular haemoglobin concentration, albumin and A/G ratio and increases inγ-GTP and creatinine were noted in females of the 1000 mg/kg group. These were considered to be unrelated to the test substance since no other changes were associated with them. A decrease in cholinesterase, which was noted in female of the 1000 mg/kg group, was not considered as the effect of the test substance since there was no corresponding changes were noted.

The other statistically significant differences, and changes in general conditions, necropsy and histopathological examinations were not considered related to administration of the test substance since there was no dose relationship and noted in the vehicle control group.

The NOEL for rats in this study was considered to be 200 mg/kg/day since increased relative liver weight was noted in males of the 1000 mg/kg group.

Conclusions:

No Observed Effect Level of the test material for rats was considered to be 200 mg/kg/day under the conditions tested. The No Observed Adverse Effect Level was considered to be 1000 mg/kg/day.

Executive summary:

A 28-day repeated-dose oral toxicity study of the test material (structural analogue source substance EC 413-110-2)

followed by a 14-day recovery test was conducted in male and female Crj: CD (SD) rats (6/sex/ group), 5 weeks of age at the start of dosing. The highest dose was set at 1000 mg/kg, and 3 lower doses at 200, 40 and 8 mg/kg. Recovery groups were separately provided for vehicle control and 1000 mg/kg groups.

There were no deaths on account of administration of the test material.

No abnormalities were noted in general conditions, body weights, food consumption, haematological examinations, blood chemical examinations, urinalysis, necropsy and histopathological examinations.

In organ weights, increased absolute and relative liver weights were noted in males of the 1000 mg/kg group.

In the recovery test, no abnormalities were noted.

In conclusion, No Observed Effect Level of the test material for rats was considered to be 200 mg/kg/day under the conditions tested.

The No Observed Adverse Effect Level was considered to be 1000 mg/kg/day.

It is proposed that this result can be used in the assessment of the target substance.

Reference 3

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Not stated.

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Principles of method if other than guideline:

3-month subacute toxicity study was conducted by oral administration to mice and rats.

GLP compliance:

no

Specific details on test material used for the study:

Sample: dibenzylidene sorbitol (D.B.S)
The sample is a white powder with pearly gloss.

Species:

rat

Strain:

other: JCL-SD strain

Sex:

male/female

Details on test animals or test system and environmental conditions:

JCL-SD strain male and female rats (40 each) were used in the study. These animals were divided into 4 groups of control group, low dose group, middle dose group and high dose group. To each group, 10 males and 10 females were assigned. In the case of rats, both males and females, 2 to 3 rats were placed in each cage.

The environmental conditions of rearing room were set at 22 - 24°C temperature, 50 - 70% humidity and 12 times of ventilation frequency. The study period was 91 days, that is, from the time these animals were 6 weeks old up to 19 weeks old.

Route of administration:

oral: feed

Details on route of administration:

Using the compounding ratio of food CE-2 for rats (CLEA Japan, Inc.) as the basis, D.B.S. was mixed at the ratios of 0.5%, 1% and 2% to prepare solid food.

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

- DIET PREPARATION :
Using the compounding ratio of food CE-2 for rats (CLEA Japan, Inc.) as the basis, D.B.S. was mixed at the ratios of 0.5%, 1% and 2% to prepare solid food and the animals were allowed free access to the food throughout the study period. CE-2 food that is the base food was given to the animals in the control group. Assuming that a rat weighing 200 g ingested about 20 g/day, the amounts of sample ingested were about 500 mg/kg in the 0.5% group, 1,000 mg/kg in the 1% group and 2,000 mg/kg in the 2% group. Tap water was given to each group of animals through water feeding bottles.

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

91 days

Frequency of treatment:

Daily

Dose / conc.:

0 other: mg/kg

Remarks:

Control

Dose / conc.:

500 other: mg/kg

Remarks:

Low dose

Dose / conc.:

1 000 other: mg/kg

Remarks:

Middle dose

Dose / conc.:

2 000 other: mg/kg

Remarks:

High dose

No. of animals per sex per dose:

10 males and 10 females per dose

Control animals:

yes, plain diet

Positive control:

None.

Observations and examinations performed and frequency:

During the study period, the general conditions of each animal including the behavior, health, hair and stools (diarrhea), etc. were observed once every day, and the body weight, food ingestion volume, water intake volume, etc. were determined once a week.

After completion of 3-month study, urinalysis was conducted. Using simultaneous urine test strips, pH, glucose, protein, occult blood and urobilinogen were qualitatively determined.

For the blood and serum tests, the animals were fasted overnight and anesthetized with pentobarbital, after which whole blood was collected from the carotid artery to determine the red blood cell count and white blood cell count (melangeur method), differential leukocyte count (usual method), hematocrit (capillary tube method), hemoglobin (cyanmethemoglobin method), plasma protein (refractometry), blood sugar (orthotoluidine borate method), S-GOT (modified Reitman-Frankel method), S-GPT (modified Reitman-Frankel method), Al-P (modified Kind-King method), BUN (urease indophenol method), serum protein fractions (rat only, electrophoresis), A/G ratio (rat only, electrophoresis).

Sacrifice and pathology:

Anatomical Test:
After the whole bloodletting, the animals were immediately autopsied and the lesions in various organs were macroscopically observed. At the same time, the liver, spleen, kidney (right and left), heart, lung, brain, pituitary, thyroid gland, adrenal gland, thymus, ovary and testis were weighed. In the case of thyroid gland and adrenal gland, both the right and left organs were determined.

Histopathological Test:
In addition to the above-mentioned organs, the stomach, small intestine, large intestine, pancreas, mesentery lymph node, submandibular gland and sternal marrow were fixed in formalin, and doubly stained with hematoxylin and eosin, after which they were microscopically investigated.

Statistics:

Of the above-mentioned test items, the mean and the standard deviation were obtained for those that can be quantitatively indicated, and any significance between the control group and dose groups was obtained.

Clinical signs:

no effects observed

Description (incidence and severity):

During the study period of 3 months, no abnormal finding was obtained from the appearance.

Mortality:

no mortality observed

Description (incidence):

During the study period of 3 months, no animal died.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Both the males and females in the dose groups did not demonstrate any significant difference from those in the control group.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

The mean food ingestion per animal a day during the study period was about 23 g in the case of males in each group, and about 15 g in the case of females in each group. Up to 1 week from the start of study, a considerable volume of spilled and dropped food was noted but after that, no difference in food ingestion was observed in comparison with those in the control group.

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

effects observed, non-treatment-related

Description (incidence and severity):

The water-intake volume per animal a day during the study period was 30 - 70 mL in the case of males and 22 - 35 mL in the case of females. Even though considerable variations were noted, there was no certain tendency between the control group and dosage of D.B.S.

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

As to the blood findings obtained from males, neutrophil increased and lymphocyte decreased in the dose groups. In addition, hemoglobin decreased in the 500 mg/kg group and hematocrit decreased in the 2,000 mg/kg group. However, no significant difference was noted in any of the blood parameters in the case of females.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

As to the serum biochemistry test, serum total protein and globulin (α1, γ) significantly increased while albumin, globulin (β), glucose and S-GPT significantly decreased in the males. However, females did not show much changes except for a tendency of increase in globulin (α1) and a tendency of decrease in S-GPT.

Urinalysis findings:

no effects observed

Description (incidence and severity):

Both the males and females showed no difference in urobilinogen and occult blood between the groups. Urine protein was ++ (100 mg/dL) in 1 animal each in the 1,000 mg/kg group and 2,000 mg/kg group but it was + (30 mg/dL) in the rest of the animals. Glucose was negative in all of them and the pH was between 6 and 8. These urinalysis findings indicated no difference between the control group and dose groups.

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Concerning the males, the measured values indicated significant differences in the weights of liver and brain of those in the 500 mg/kg group, and in that of the liver in those in the 1,000 mg/kg group and 2,000 mg/kg group. As to the body weight ratio, there was a significant difference in the liver of animals in the 500 mg/kg group and 2,000 mg/kg group. Concerning the females, the measured values indicated significant differences in the liver of those in the 500 mg/kg group, 1,000 mg/kg group and 2,000 mg/kg group, and in the ovary of those in the 2,000 mg/kg group. Concerning the body weight ratio, a significant difference was noted in the liver and ovary of those in the 2,000 mg/kg group. As to the liver, both the measured value and body weight ratio were higher in those in the dose groups than those in the control group both in the case of males and females, and the values were especially the highest in the 2,000 mg/kg group. As to other organs, there was no difference between the control group and dose groups.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

As to the males, clear hepatic lobule was noted in 1 animal in the control group and in 3 animals in the 2,000 mg/kg group. Furthermore, a cartilage-like calculus was observed in the 2,000 mg/kg group. As to the females, localized congestion in the lung was noted in 2 animals in the control group, 1 in the 500 mg/kg group and 1 in the 2,000mg/kg group. The lesions in the liver included white spots in 1 animal in the control group and slight hypertrophy in 1 animal in the 500 mg/kg group. As to the lesion in the kidney, mild renal edema occurred in the right and left kidneys in the 2,000 mg/kg group but this is a symptom frequently observed in SD strain rats.

Among the lesions cited in the above, there was none that is considered to have occurred as an influence of D.B.S. administration.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

As to the control group, calcification in the distal kidney tubule was noted in 1 female, while nodulation due to hyperplasia of bronchial mucosal epithelium of lung was found in 1 male and a node formed by the fibrosis and lymphocyte infiltration in the myocardial layer in another male. Other than that, unstructured or sometimes granular cast-like substance that was slightly stained in eosin was observed in the proximal tubule close to the renal cortex of several males and females (photo 9).

As to the dose groups, interstitium hypertrophy of lung, sometimes accompanied by lymphocyte infiltration was noted in several animals in each group. One hyperplasia-like node occurred in the exocrine pancreas in 1 animal in the 2,000 mg/kg group, and slightly more cells in the spleen indicated hemosiderin phagocytosis . The cast-like substance in the renal tubule was observed both in males and females of dose groups to a degree that is not much different from that noted in the control group. Other than that, no special change assumed to be attributable to D.B.S. administration was observed in the liver, stomach, small intestine, large intestine, testis, uterus, ovary, etc.

Histopathological findings: neoplastic:

not examined

Details on results:

Firstly, the volume of D.B.S. ingested in relation to the food ingestion volume: In the case of rats, the doses of D.B.S. were set at 500, 1,000 and 2,000 mg/kg by assuming that they consumed 20 g of food each day. Actually, the males ingested more than the prospected amount and the females slightly less than that. In comparison with the rats in the control group, nothing abnormal was observed in the general symptoms, food ingestion volume, water-intake volume and urinalysis results in those in the dose groups.

As to the blood test and serum biochemistry test In the case of rats, changes were noted in the neutrophil and lymphocyte in the males but there was no certain relation to the dose and nothing abnormal was noted in the leukocyte count. Albumin decreased and globulin (α1, α2, β, γ) increased, which led to a decrease in A/G ratio, in the 1,000 and 2,000 mg/kg groups. It is reported that GOT, GPT, AI-P, BUN, etc. demonstrate marked changes depending on the degree of disorder in the liver and kidney, etc. caused by the toxicity of drug. The GOT in the 1,000 and 2,000 mg/kg groups tended to increase also in this study, while GPT and urea nitrogen tended to decrease both in the males and females. However, these tendencies seemed to have been related to the high measured value of liver and its ratio to body weight in the high dose group. In spite of that, hardly any change was observed in the liver of rats from the histopathological point, indicating that the degree of influence remained within the range of physiological variation.

As to the autopsy findings, clear hepatic lobule was observed in the liver of some animals, and the histological test indicated hypertrophy of liver cells in 3 male mice in the 3,200 mg/kg group. However, these findings were all observed in a part of tissue examined by microscope. Furthermore, these changes were not observed in the female mice, and male and female rats in the maximum dose group. Nor such changes were noted in the mice and rats in other dose groups. Accordingly, it is not possible to conclude the above-mentioned finding as an influence of D.B.S.

The above results indicated that no markedly abnormal findings assumed to be expressed by the administration of dibenzyliden sorbitol were obtained from any of the tests.

Key result

Dose descriptor:

NOAEL

Remarks:

(rat)

Effect level:

2 000 other: mg/kg

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: The maximum dose in the study that does not demonstrate toxicity was 2,000 mg/kg/day for the male and female rats.

Key result

Critical effects observed:

no

Conclusions:

A 3-month subacute toxicity study by oral administration of D.B.S. indicated that dibenzyliden sorbitol was very stable and had hardly any toxicity. The maximum dose that did not demonstrate toxicity in the study was 2,000 mg/kg/day for the male and female rats.

Executive summary:

A 3-month subacute study of dibenzyliden sorbitol was conducted in mice and rats, and the following results were obtained.

 

No special changes were noted in the general symptoms, food ingestion volume, water-intake volume, body weight change, urinalysis findings, etc. The blood and serum biochemistry tests indicated significant differences in some parameters both in mice and rats but all these changes were within the physiological range of variation and indicated no relation to the administration of D.B.S. 

 

Macroscopic autopsy findings detected some lesions in the kidney, liver, lung and bladder, etc. of some animals but there was no change specifically observed in the dose groups. As to the organ weights, the weight of liver tended to increase slightly but there was no other tendency that was commonly observed in the males and females of mice and rats and that demonstrated a proportionally certain relation to the dose in the remaining organs.

 

As to the histopathological findings, some localized abnormal findings were obtained from the lung, kidney, heart, liver, pancreas and adrenal gland but all of these events occurred as single episodes and there was no specific change to indicate the correlation to the administration of D.B.S.

 

The maximum dose that did not demonstrate  toxicity in the study was 2,000 mg/kg/day for the male and female rats.

Reference 4

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Not stated.

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Principles of method if other than guideline:

3-month subacute toxicity study was conducted by oral administration to mice and rats.

GLP compliance:

no

Specific details on test material used for the study:

Sample: dibenzylidene sorbitol (D.B.S)
The sample is a white powder with pearly gloss.

Species:

mouse

Strain:

other: JCL-ICR

Sex:

male/female

Details on test animals or test system and environmental conditions:

JCL-ICR strain male and female mice (40 each) were used in the study. These animals were divided into 4 groups of control group, low dose group, middle dose group and high dose group. To each group, 10 males and 10 females were assigned. The male mice were placed in individual cages so as to avoid quarrelling between them while 3 to 4 females were placed in each cage.

The environmental conditions of rearing room were set at 22 - 24°C temperature, 50 - 70% humidity and 12 times of ventilation frequency. The study period was 91 days, that is, from the time these animals were 6 weeks old up to 19 weeks old.

Route of administration:

oral: feed

Details on route of administration:

Using the compounding ratio of food CE-2 for mice (CLEA Japan, Inc.) as the basis, D.B.S. was mixed at the ratios of 0.5%, 1% and 2% to prepare solid food.

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

- DIET PREPARATION :
Using the compounding ratio of food CE-2 for mice and rats (CLEA Japan, Inc.) as the basis, D.B.S. was mixed at the ratios of 0.5%, 1% and 2% to prepare solid food and the animals were allowed free access to the food throughout the study period. CE-2 food that is the base food was given to the animals in the control group. Assuming that a mouse weighing 30 g ingested 5 g/day, the amounts of D.B.S. administered per day through the food became about 800 mg/kg in the 0.5% group, 1,600 mg/kg in the 1% group and 3,200 mg/kg in the 2% group.

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

91 days

Frequency of treatment:

Daily

Dose / conc.:

0 other: mg/kg

Remarks:

Control

Dose / conc.:

800 other: mg/kg

Remarks:

Low dose

Dose / conc.:

1 600 other: mg/kg

Remarks:

Middle dose

Dose / conc.:

3 200 other: mg/kg

Remarks:

High dose

No. of animals per sex per dose:

10 males and 10 females per dose

Control animals:

yes, plain diet

Positive control:

None.

Observations and examinations performed and frequency:

During the study period, the general conditions of each animal including the behavior, health, hair and stools (diarrhea), etc. were observed once every day, and the body weight, food ingestion volume, water intake volume, etc. were determined once a week.

After completion of 3-month study, urinalysis was conducted. Using simultaneous urine test strips, pH, glucose, protein, occult blood and urobilinogen were qualitatively determined.

For the blood and serum tests, the animals were fasted overnight and anesthetized with pentobarbital, after which whole blood was collected from the carotid artery to determine the red blood cell count and white blood cell count (melangeur method), differential leukocyte count (usual method), hematocrit (capillary tube method), hemoglobin (cyanmethemoglobin method), plasma protein (refractometry), blood sugar (orthotoluidine borate method), S-GOT (modified Reitman-Frankel method), S-GPT (modified Reitman-Frankel method), Al-P (modified Kind-King method), BUN (urease indophenol method), serum protein fractions (rat only, electrophoresis), A/G ratio (rat only, electrophoresis).

Sacrifice and pathology:

Anatomical Test:
After the whole bloodletting, the animals were immediately autopsied and the lesions in various organs were macroscopically observed. At the same time, the liver, spleen, kidney (right and left), heart, lung, brain, pituitary, thyroid gland, adrenal gland, thymus, ovary and testis were weighed. In the case of thyroid gland and adrenal gland, both the right and left organs were determined.

Histopathological Test:
In addition to the above-mentioned organs, the stomach, small intestine, large intestine, pancreas, mesentery lymph node, submandibular gland and sternal marrow were fixed in formalin, and doubly stained with hematoxylin and eosin, after which they were microscopically investigated.

Statistics:

Of the above-mentioned test items, the mean and the standard deviation were obtained for those that can be quantitatively indicated, and any significance between the control group and dose groups was obtained.

Clinical signs:

no effects observed

Description (incidence and severity):

During the study period, no particular abnormality was noted in the behavior, health, hair and stools of mice.

Mortality:

no mortality observed

Description (incidence):

During the study period of 3 months, no animal died.

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

A tentative decrease in body weight occurred in the males in Week 4. The body weight in the dose groups tended to be lower than that in the control group on the whole, and there was a significant difference in Week 6 in the 3,200 mg/kg group. However, no significant difference was observed in other weeks. Females in the dose groups did not demonstrate any significant difference from those in the control group.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

As to the mean food ingestion per animal a day during the study period, males in the control group, 800 mg/kg group, 1,600 mg/kg group, 3,200 mg/kg group ingested 5.4 g, 6.4 g, 6.6 g and 7.4 g respectively, indicating that those taking the food with more D.B.S. content consumed more food. Females ingested, 4.3 g, 7.4 g, 8.4 g and 5.9 g respectively. Those in the dose groups tended to ingest more food.
In the measurement of food ingestion, the food spilled and dropped was collected as much as possible before measurement. However, the food in the dose groups was fragile and easily crushed, which increased the volume spilled and dropped. As a result, the food ingestion in the dose groups showed an increase over the food consumed in the control group. Prior to the study, the dose of D.B.S. was determined by assuming that the food ingested per mouse per day was 5 g. It is estimated that each animal ingested about 5 g of food a day also in this study.

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

effects observed, non-treatment-related

Description (incidence and severity):

The drinking water consumed per animal a day during the study period was between 4 and 10 mL in each group. No significant difference was observed between the dose groups in the case of males. In general, females in the dose groups tended to drink less water in comparison with the control group.

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

As to the males, significant differences were noted in the neutrophil and the lymphocyte in the 1,600 mg/kg group, and the alkaline phosphatase in the 800 mg/kg group. As to the females, significant differences were noted in the hematocrit and the hemoglobin in the 3,200 mg/kg group, and the alkaline phosphatase in the 800 mg/kg group. The value tended to be higher in the dose groups than in the control group. No change was noted in the remaining parameters..

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

No change was noted in the remaining parameters..

Urinalysis findings:

no effects observed

Description (incidence and severity):

A trace amount of urobilinogen was detected from all the animals but occult blood was negative in all of them. Urine protein was ++ (100 mg/dL) in some of the animals but it was + (30 mg/dL) in many of them. In this regard, no particular relation to the dose was observed. Glucose was negative in all of them. The pH was between 6 and 8, indicating no particular abnormality.

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

Concerning the males, the measured values indicated significant differences in the thymus in the 800 mg/kg group, testis (right) in the 1,600 mg/kg group, and heart and thyroid gland in the 3,200 mg/kg group, but the body weight ratios indicated significant difference in the weight of liver and thymus in the 800 mg/kg group, and liver in the 3,200 mg/kg group. As to the females, the measured values indicated significant differences in the liver, kidney (right and left), heart, lung, thyroid gland and adrenal gland in the 800 mg/kg group, in the liver in the 1,600 mg/kg group, and in the liver, kidney (right and left), lung, brain and adrenal gland in the 3,200 mg/kg group but significant differences were found only in the liver in the 800 mg/kg and 1,600 mg/kg groups, and in the liver, spleen and kidney (right) in the 3,200 mg/kg group as to the body weight ratio. However, no certain correlation that is proportional to the dosage was observed even in those organs that demonstrated significant differences.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

As to the males, somewhat clear hepatic lobule was noted in one animal each in the 800, 1,600 and 3,200 mg/kg groups. However, no lesion was observed in any of the organs of the remaining animals. No noteworthy lesion was noted in the females in any of the groups.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

As to the control group, a fibrotic nodule due to the lymphocyte infiltration was noted in the liver of 1 female. In addition to 1 renal cyst in 1 male, cast-like substance was noted in the renal tubule of both males and females. As to the females of dose groups, nodule due to cell infiltration mainly the lymphocyte was observed in a part of the liver of 1 animal, fatty degeneration in only a limited part of cardiac muscle in another, and fat deposition in the border of adrenal gland cortex and medulla in another in the 3,200 mg/kg group were mainly the changes noted. As to the males, adenoma-like image in the lung in 1 animal, and hypertrophy of liver cells and nucleus around the hepatic lobule were noted in 3 animals. Other than the above, the cast-like substance in the renal tubule was also noted in the males and females to the same degree as that observed in the control group.
The cast-like substance in the kidney, interstitium hypertrophy of lung or lymphocyte infiltration were observed in several animals also in the 800 mg/kg and 1,600 mg/kg groups.

Histopathological findings: neoplastic:

not examined

Details on results:

Firstly, the volume of D.B.S. ingested in relation to the food ingestion volume: In the case of mice, the dose was set by assuming that they consumed 5 g of food each day. In this regard, the mice in each group mostly ingested 5 g or more food. In other words, those in the dose groups ingested 800 mg/kg, 1,600 mg/kg or 3,200 mg/kg of D.B.S. per day.

In comparison with the mice in the control group, nothing abnormal was observed in the general symptoms, food ingestion volume, water-intake volume and urinalysis results in those in the dose groups. As to the body weight change in mice, a tentative decrease was observed in Week 4 and 6. However, since this tendency was noted in each group, the change is not considered as an influence of administration of this substance.

As to the blood test and serum biochemistry test in mice, significant differences from the control group were observed in the values of neutrophil, lymphocyte and alkaline phosphatase in the males, and hematocrit, hemoglobin and alkaline phosphatase, etc. in the females. However, none of these changes exceeded the physiologically normal range.

As to the organ weights, the thyroid gland, adrenal gland, thymus, etc. demonstrated significant differences in some of the animals in comparison with those in the control group. However, in view of the absence of any change common to males and females and that of any certain relation to the dose, these changes are not considered as an influence of this substance.

As to the autopsy findings, clear hepatic lobule was observed in the liver of some animals, and the histological test indicated hypertrophy of liver cells in 3 male mice in the 3,200 mg/kg group. However, these findings were all observed in a part of tissue examined by microscope. Furthermore, these changes were not observed in the female mice, and male and female rats in the maximum dose group. Nor such changes were noted in the mice and rats in other dose groups. Accordingly, it is not possible to conclude the above-mentioned finding as an influence of D.B.S. Other than the above, the results of histopathological examination of circulation system (heart), respiration system (lung), digestive system (salivary gland, liver, pancreas, stomach, small intestine, and large intestine), urogenital system (kidney, bladder, testis, uterus, and ovary), hematopoietic system (bone marrow, spleen, and lymph node) and endocrine system (pancreas and adrenal gland) indicated mild histopathological changes both in the control group and dose groups. However, it was not possible to determine that these changes were the changes specifically caused by the administration of D.B.S.

The above results indicated that no markedly abnormal findings assumed to be expressed by the administration of dibenzyliden sorbitol were obtained from any of the tests.

Key result

Dose descriptor:

NOAEL

Remarks:

(mouse)

Effect level:

3 200 other: mg/kg

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: The maximum dose in the study that does not demonstrate toxicity was 3,200 mg/kg/day for the male and female mice

Key result

Critical effects observed:

no

Conclusions:

A 3-month subacute toxicity study by oral administration of D.B.S. indicated that dibenzyliden sorbitol was very stable and had hardly any toxicity. The maximum dose that did not demonstrate toxicity in the study was 3,200 mg/kg/day for the male and female mice.

Executive summary:

A 3-month subacute study of dibenzyliden sorbitol was conducted in mice and rats, and the following results were obtained.

 

No special changes were noted in the general symptoms, food ingestion volume, water-intake volume, body weight change, urinalysis findings, etc. The blood and serum biochemistry tests indicated significant differences in some parameters both in mice and rats but all these changes were within the physiological range of variation and indicated no relation to the administration of D.B.S. 

 

Macroscopic autopsy findings detected some lesions in the kidney, liver, lung and bladder, etc. of some animals but there was no change specifically observed in the dose groups. As to the organ weights, the weight of liver tended to increase slightly but there was no other tendency that was commonly observed in the males and females of mice and rats and that demonstrated a proportionally certain relation to the dose in the remaining organs.

 

As to the histopathological findings, some localized abnormal findings were obtained from the lung, kidney, heart, liver, pancreas and adrenal gland but all of these events occurred as single episodes and there was no specific change to indicate the correlation to the administration of D.B.S.

 

The maximum dose that did not demonstrate  toxicity in the study was 3,200 mg/kg/day for the male and female mice.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

No adverse effects observed in repeated dose toxicity studies conducted on the substance and a structurally similar substance.

Justification for classification or non-classification

Based on the available study data on the substance itself and on structurally similar substance, classification for Specific Target Organ Toxicity - Repeated Exposure (STOT-RE), is not justified.