Registration Dossier

Administrative data

Description of key information

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

- 13-week oral toxicity test (oral feed) in rats and mice on target substance (EC 406-176-9).

- reading across the results of a 28-day repeated dose toxicity study conducted on a structurally similar analogue substance (EC 413-110-2).

13-week oral toxicity test (oral feed) in rats and mice on target substance (EC 406-176-9).

In a 13-week repeated dose toxicity oral feed study, rats were treated at concentrations up to 50000 ppm. No significant adverse toxic effects were observed at 50000 ppm in the parameters examined.

The NOAEL is considered to be 50000 ppm; equivalent to 3020 mg/kg bw/day in males and 3240 mg/kg bw/day in females.

In a 13-week repeated dose toxicity oral feed study, mice were treated at concentrations up to 50000 ppm. No significant adverse toxic effects were observed at 50000 ppm in the parameters examined.

The NOAEL is considered to be 50000 ppm; equivalent to 7280 mg/kg bw/day in males and 10710 mg/kg bw/day in females.

28 -day repeated dose toxicity study:

Source Substance; 1,3:2,4-bis-O-(3,4-dimethylbenzylidene)-D-glucitol (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.

It is proposed that this result can be used in the assessment of the target substance (EC 406-176-9)

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1981
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study without detailed documentation
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Route of administration:
oral: feed
Details on route of administration:
Method of administration: Diet
Vehicle:
unchanged (no vehicle)
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Treated diet available throughout study
Dose / conc.:
0 ppm
Remarks:
Control
Dose / conc.:
5 000 ppm
Dose / conc.:
15 000 ppm
Dose / conc.:
50 000 ppm
No. of animals per sex per dose:
20 males and 20 females per dose
Control animals:
yes, plain diet
Positive control:
No
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
BODY WEIGHT: Yes
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
FOOD EFFICIENCY: Yes
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
OPHTHALMOSCOPIC EXAMINATION: No
HAEMATOLOGY: Yes
CLINICAL CHEMISTRY: Yes
URINALYSIS: Yes
NEUROBEHAVIOURAL EXAMINATION: Not specified
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
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:
no effects observed
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Details on results:
CLINICAL OBSERVATIONS:
No deaths occurred nor abnormal symtoms in any of treated or control groups throughout the study.
There were also no abnormal symtoms in the general conditions of the satellite group.
Appearance, behaviour, mortality, body weight gain, food consumption, water intake and food efficiency of rats treated were not affected throughout the study.

LABORATORY FINDINGS:
In the interim examination of the satellite group in weeks 5, 9 and 13, there were no abnormalities in urinanalysis, hematology and clinical chemistry parameters attrtibuted to the test material.

In the terminal examination at the end of the treatment period of 13 weeks, there were no abnormalities in urinanalysis, hematology, clinical chemistry, necropsy, organ weight and histopathology parameters attributed to the test material.

EFFCTS IN ORGANS:
There were no significant changes between the treated and control groups or absolute and relative organ weight.

In absolute organ weight, pituitary weight of females fed 15000 ppm, thymic gland, heart, lung, liver and kidney of females fed 50000 ppm increased significantly when compared with those of the control group. In relative organ weight, thymic gland and kidney decreased in females fed 5000 ppm and relative brain weight decreased in females fed 50000 ppm.

Histopathological changes were seen in the pituitary, thyroids, thymus, liver, kidney, stomach, duodenum, urinary bladder, testis, prostate and skeletal muscle of treated or control animals. These abnormalities were spontaneous or accidental changes, normal for species and ages of the test animals.












Dose descriptor:
NOAEL
Effect level:
50 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Dose at which no toxic effects were observed
Dose descriptor:
NOAEL
Remarks:
Equivalent dose based on food intake/bodyweight
Effect level:
3 020 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: Dose at which no toxic effects were observed
Dose descriptor:
NOAEL
Remarks:
Equivalent dose based on food intake/bodyweight
Effect level:
3 240 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: Dose at which no toxic effects were observed
Critical effects observed:
no

Dose/concentration at which no toxic effects were observed: 50000 ppm; 3.02 g/kg/day (males); 3.24 g/kg/day (females)

Conclusions:
In a 13-week repeated dose toxicity oral feed study, rats were treated at concentrations up to 50000 ppm. No significant adverse toxic effects were observed at 50000 ppm in the parameters examined.
The NOAEL is considered to be 50000 ppm; equivalent to 3020 mg/kg bw/day in males and 3240 mg/kg bw/day in females.
Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1981
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study without detailed documentation
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
GLP compliance:
not specified
Limit test:
no
Species:
mouse
Strain:
CD-1
Sex:
male/female
Route of administration:
oral: feed
Details on route of administration:
Method of administration: Diet
Vehicle:
unchanged (no vehicle)
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Treated diet available throughout study
Dose / conc.:
0 ppm
Remarks:
Control
Dose / conc.:
5 000 ppm
Dose / conc.:
15 000 ppm
Dose / conc.:
50 000 ppm
No. of animals per sex per dose:
20 males and 20 females per dose
Control animals:
yes, plain diet
Positive control:
No.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
BODY WEIGHT: Yes
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
FOOD EFFICIENCY: Yes
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
OPHTHALMOSCOPIC EXAMINATION: No
HAEMATOLOGY: Yes
CLINICAL CHEMISTRY: Yes
URINALYSIS: Yes
NEUROBEHAVIOURAL EXAMINATION: Not specified
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
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:
no effects observed
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Details on results:
CLINICAL OBSERVATIONS:
No deaths occurred nor abnormal symtoms in any of treated or control groups througout the study.
Appearance, behaviour, mortality, body weight gain, food consumption, water intake and food efficiency of rats treated were not affected throughout the study.

LABORATORY FINDINGS:
In the interim examination of the satellite group in week 5t here were no abnormalities in urinanalysis, hematology and clinical chemistry parameters attrtibuted to the test material.

In the terminal examination at the end of the treatment period of 13 weeks, there were no abnormalities in urinanalysis, hematology, clinical chemistry, necropsy, organ weight and histopathology parameters attributed to the test material.

EFFECTS IN ORGANS:
The increased absolute and and relative kidney weight was recorded in males fed 50000 ppm and in females fed 15000 ppm. Absolute kidney weight also increased in females fed 50000 ppm. The decreased absolute and relative spleen weight was recorded in males fed 5000 ppm. Increased absolute pituitary and thymus weight and decreased relative testis weight were recorded in males fed 5000 ppm.
These changes in the 5000 ppm group were not dose-related changes. There were no other significant changes in organ weight parameters in treated groups.

Histopatholigical changes were seen in the pituitary, thymus, liver, kidney, adrenal, pancreas, stomach and eye of treated and control animals. These abnormalites were thought to be spontaneous or accidental changes.








Dose descriptor:
NOAEL
Effect level:
50 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Dose at which no toxic effects were observed
Dose descriptor:
NOAEL
Remarks:
Equivalent dose based on food intake/bodyweight
Effect level:
7 280 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: Dose at which no toxic effects were observed
Dose descriptor:
NOAEL
Remarks:
Equivalent dose based on food intake/bodyweight
Effect level:
10 710 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: Dose at which no toxic effects were observed
Critical effects observed:
no

Dose/concentration at which no toxic effects were observed: 50000 ppm; 7.28 g/kg/day (males); 10.71 g/kg/day (females)

Conclusions:
In a 13-week repeated dose toxicity oral feed study, mice were treated at concentrations up to 50000 ppm. No significant adverse toxic effects were observed at 50000 ppm in the parameters examined.
The NOAEL is considered to be 50000 ppm; equivalent to 7280 mg/kg bw/day in males and 10710 mg/kg bw/day in females.
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:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Qualifier:
according to
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. certificate)
Limit test:
no
Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals 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. 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 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.

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:
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 chemical:
1,3:2,4-bis-O-(3,4-dimethylbenzylidene)-D-glucitol (EC 413-110-2, CAS 135861-56-2)

Target chemical:
2,6-bis(4-ethylphenyl)perhydro-1,3,5,7-tetraoxanaphth-4-ylethane-1,2-diol (EC 406-176-9, CAS 79072-96-1)

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 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 'Read-across justification to support the REACH registration of EC 406-176-9' document attached in section 13.
Reason / purpose:
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 (EC 406-176-9).

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.