Registration Dossier

Administrative data

Description of key information

NOAEL was in the dose range of 190-800 mg/kg bw for test chemical when exposed orally to rodent .

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
chronic toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Remarks:
Experimental data of read across substances
Justification for type of information:
Weight of evidence approach based on structurally similar chemicals
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
other: as below
Principles of method if other than guideline:
WoE report is based on two repeated dose oral toxicity studies
1. Repeated dose oral toxicity study was performed to determine the toxic nature of the test chemical
2. 80 weeks chronic repeated dose oral toxicity study of test chemical was performed to determine its chronic toxic nature
GLP compliance:
not specified
Limit test:
no
Species:
other: 1. Rat, 2. mice
Strain:
other: 1. Tif:RAIf, SPF, CD
Sex:
male/female
Details on test animals and environmental conditions:
2. - Source: SPF breeding colony
- Weight at study initiation: Male- 21-30 g and female - 17-25 g
- Fasting period before study: no
- Housing: housed in cages of 15
- Diet (e.g. ad libitum): ground Oxoid pasteurized breeding diet; ad libitum
- Water (e.g. ad libitum): water ad libitum
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21± 1°C
- Humidity (%):50-60%
Route of administration:
oral: feed
Vehicle:
other: 1. Feed, 2. basic diet
Details on oral exposure:
1. PREPARATION OF DOSING SOLUTIONS: The test chemical was mixed with feed at dose level of 0, 500, 5000, or 50,000 ppm (19, 190, and 2300 mg/kg/day in males and 21, 226, and 2620 mg/kg/day in females).

DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data

VEHICLE
- Justification for use and choice of vehicle (if other than water): Feed
- Concentration in vehicle: 0, 500, 5000, or 50,000 ppm (19, 190, and 2300 mg/kg/day in males and 21,
226, and 2620 mg/kg/day in females)
- Amount of vehicle (if gavage): No data
- Lot/batch no. (if required): No data
- Purity: No data

2. PREPARATION OF DOSING SOLUTIONS: The test chemical was mixed with feed at dose levels of 0, 100, 200, 400, 800 mg/l (0, 0.2, 0.4, 0.8 or 1.6%)

DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): Oxoid pasteurized breeding diet
- Storage temperature of food: No data

VEHICLE
- Justification for use and choice of vehicle (if other than water): Oxoid pasteurized breeding diet
- Concentration in vehicle: 0, 100, 200, 400, 800 mg/l (0, 0.2, 0.4, 0.8 or 1.6%)
- Amount of vehicle (if gavage): No data
- Lot/batch no. (if required): No data
- Purity: No data
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
1. Chemical analysis of the feed was done
Duration of treatment / exposure:
1. Chronic
2. 80 weeks
Frequency of treatment:
Daily
Remarks:
1. 0, 500, 5000, or 50,000 ppm (19, 190, and 2300 mg/kg/day in males and 21, 226, and 2620 mg/kg/day in females)
Remarks:
2. 0, 100, 200, 400, 800 mg/l (0, 0.2, 0.4, 0.8 or 1.6%)
No. of animals per sex per dose:
2. Control group:60 male and 60 females
Test group:
100 mg/Kg: 30 male and 30 females
200 mg/Kg: 30 male and 30 females
400 mg/Kg: 30 male and 30 females
800 mg/Kg: 30 male and 30 females
Control animals:
yes, concurrent vehicle
Observations and examinations performed and frequency:
1. CAGE SIDE OBSERVATIONS: Yes
- Time schedule: No data
- Cage side observations checked in table [No.?] were included. Mortality, appearance and behavior
DETAILED CLINICAL OBSERVATIONS: No data
- Time schedule: No data
BODY WEIGHT: Yes
- Time schedule for examinations: No data
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/
kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain
data: No data
FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averag
es from the consumption and body weight gain data: No data
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: No data
OPHTHALMOSCOPIC EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
HAEMATOLOGY: Yes
- Time schedule for collection of blood: No data
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. No data
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. No data
URINALYSIS: No data
- Time schedule for collection of urine: No data
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.?] were examined. No data
NEUROBEHAVIOURAL EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data

2. CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Continued surveillance
- Cage side observations checked in table [No.?] were included. Any abnormalities in condition and behav
ior
DETAILED CLINICAL OBSERVATIONS: No data
- Time schedule: No data
BODY WEIGHT: Yes
- Time schedule for examinations: At 4 weeks interval throughout the study
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/
kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain
data: No data
FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted av
erages from the consumption and body weight gain data: No data
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations: No data
OPHTHALMOSCOPIC EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
HAEMATOLOGY: Yes
- Time schedule for collection of blood: wk 13, 26 and 52 and from all surviving mice at wk 80
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 10/sex from control group and animals fed 400 and 800 mg/Kg
- Parameters checked in table [No.?] were examined. Haemoglobin concentration and packed cell volume
and counts were made of erythrocytes and total leucocytes. Differential leucocyte counts and reticulocyte
counts were made on the blood from the mice fed on the control diet and from those given diet containing
1.6% Sunset Yellow FCF at wk 26, 52 and 80, and reticulocyte counts were also carried out for these
groups at wk 13.
CLINICAL CHEMISTRY: No data
- Time schedule for collection of blood: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. No data
URINALYSIS: No data
- Time schedule for collection of urine: No data
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.?] were examined. No data
NEUROBEHAVIOURAL EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data
IMMUNOLOGY: Yes / No / Not specified
- Time schedule for examinations:
- How many animals:
- Dose groups that were examined:
- Parameters checked in table [No.?] were examined.
Sacrifice and pathology:
1.GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

2. GROSS PATHOLOGY: Yes, At autopsy, all macroscopic abnormalities were noted and the brain, heart, liver, kidneys, spleen, stomach, small intestine, caecum and testes were weighed.

HISTOPATHOLOGY: Yes, the brain, heart, liver, kidneys, spleen, stomach, small intestine, caecum and testes together with salivary gland, thyroid, adrenals, lymph nodes, pancreas, pituitary, ovaries, uterus, urinary bladder, lungs, colon, rectum, spinal cord, skeletal muscle and any other tissue that appeared abnormal were preserved in 10% buffered formalin. Paraffin-wax sections of these tissues were stained with haemotoxylin and eosin. All tissues from the control mice and those fed 800 mg/Kg Sunset Yellow FCF were examined microscopically, while at the lower dietary levels examination was confined to the liver and kidney together with any tissue seen to be abnormal at autopsy.
Other examinations:
2. Behavior and were weighed at 4-wk intervals throughout the study. During the first half of the study it was noticed that there was a tendency for the male mice to fight. Bite lesions of the anogenital region were particularly frequent and these were associated with obstructions of the urinary tract. To avoid further fighting, all the mice were caged individually from month 8.
Clinical signs:
no effects observed
Description (incidence and severity):
1. The test substance did not affect appearance and behavior and was similar in all groups of test.
Mortality:
no mortality observed
Description (incidence):
1. Mortality was unaffected by the test chemical
2. There were deaths in all groups during the study and there was no relationship between the number of deaths at any time and the dietary intake of test chemical.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
1. 10% decrease in body weight were observed in 2300 mg/Kg/day dose group in males and 2620 mg/Kg/ day dose group in females.
2. The terminal body weights and the body-weight gains were similar in all groups.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
1. Increased food consumption was observed in 2300 mg/Kg/day dose group in males and 2620 mg/Kg/day dose group in females.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
effects observed, treatment-related
Description (incidence and severity):
1.Water consumption showed a dose-related increase in the two highest dose groups (190 and 2300 mg/ Kg/day and 226 and 2620 mg/Kg/day)
Haematological findings:
no effects observed
Description (incidence and severity):
1. Hematology did not show treatment-related effects.
2.No evidence of any haematological adverse effect due to the administration of test chemical
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
1. Clinical blood chemistry parameters did not show treatment-related effects.
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
1. An increase in urine output was observed in the two highest dose groups (190 and 2300 mg/Kg/day and 226 and 2620 mg/Kg/day)
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
1. In the high dose group, significantly increased organ-to-body weight ratios for liver (p<0.01) were seen in males (2300 mg/Kg/day), and in males and females for kidney (p<0.05) (2300 mg/Kg/day for males and 2620 mg/Kg/day for females). The test substance did not adversely affect other parameters.
2. No significant differences between the organ weights or relative organ weights of the test groups and the corresponding controls.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
1. Macroscopic examinations at necropsy revealed an increased incidence of pancreateic nodules and masses in the 2300 mg/kg/day males and 2620 mg/Kg/day females.
2. Distension of the bladder, frequently associated with hydroureter and hydronephosis was noted. Pus and hard protein deposits were found in the bladders of many of the mice and in a number of cases it was possible to show that an obstruction had occurred in the urethra in the region of the bulbo-urethral complex. After the male mice were put into individual cages, there was a marked reduction in the number showing this syndrome, although isolated cases of urinary retention were seen throughout the study.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
1. Histopathology revealed statistically significant test article-related neoplastic effects to the pancreas of high dose males and females. Nodular hyperplasia of the exocrine pancreas was observed in 51 of 78 (p<0.001) high dose (2300 mg/Kg/day) males versus 6 of 78 control group males, and in 10 of 79 (p<0 .05) high dose (2620 mg/Kg/day) females versus 1 of 78 control group females. Other non-neoplastic lesions seen in the dosed animals were not related to treatment with FWA-5.
2. No differences between treated and control mice in the incidence or severity of the lesions seen. There as an unusually high incidence of pyelonephritis and chronic inflammation of the bladder and urethra, but these lesions occurred mainly in the male mice killed because of urinary retention in the early stages of the study.
Histopathological findings: neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
1. Exocrine pancreas adenoma, a benign tumor, was observed in males of the 5,000 and 50,000 ppm dose groups at an incidence of 2/79 and 18/78 (p<0.05), respectively, compared with 0/78 in control. Pancreatic carcinoma, a malignant tumor, was also observed in 2/78 high-dose males and although not statistically significantly increased, the rarity of this tumor and the treatment-related finding of adenomas in this group suggest the carcinomas could be related to the treatment. In females, adenomas in exocrinepancreas occurred only in the high dose (2620 mg/Kg/day) group (2/79) and were within the historical control range; carcinomas were not observed in any of the females.
2. Of the malignant tumours found, lymphomas and reticulum-cell neoplasms occurred in treated and control mice. In addition a splenic haemangiosarcoma and a mammary carcinoma were found in the female mice fed on the diet containing the lowest level of test chemical (100 mg/Kg). A single tumour of the Harderian gland was found in a female from the group given the 1.6% level. The commonest benign tumours were pulmonary adenomas and benign cysts in the ovary, found with a similar frequency in treated and control mice. The only other tumour occurring in treated mice without a similar finding in the controls was a granulosa-cell tumour of the ovary in a mouse given 800 mg/Kg.
Other effects:
not specified
Dose descriptor:
NOAEL
Remarks:
1
Effect level:
190 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
water consumption and compound intake
haematology
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
histopathology: neoplastic
Remarks on result:
other: No effect observed
Dose descriptor:
NOAEL
Remarks:
1
Effect level:
226 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
haematology
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
histopathology: neoplastic
Remarks on result:
other: No effect observed
Dose descriptor:
NOAEL
Remarks:
2.
Effect level:
800 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
haematology
organ weights and organ / body weight ratios
histopathology: non-neoplastic
histopathology: neoplastic
Remarks on result:
other: No effect observed
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Conclusions:
NOAEL was in the rang of 190-800 mg/kg bw for test chemical when exposed orally.
Executive summary:

Data available for target and the read across chemicals was reviewed to determine the repeated dose oral toxicity of 2 ,2'-(ethene-1,2-diyldi-4,1-phenylene)bis(1,3-benzoxazole) (CAS no 1533-45-5). The studies are as mentioned below:

Study 1:

Repeated dose oral toxicity study was performed to determine the toxic nature of the test chemical. The study was performed using male and female Tif:RAIf, SPF strain rats. The test chemical was mixed with feed at dose levels of 0, 500, 5000, or 50,000 ppm (19, 190, and 2300 mg/kg/day in males and 21, 226, and 2620 mg/kg/day in females). The test substance did not affect appearance and behavior and was similar in all groups of test. Mortality was unaffected by the test chemical. 10% decrease in body weight was observed in 2300 mg/Kg/day dose group in males and 2620 mg/Kg/day dose group in females. Increased food consumption was observed in 2300 mg/Kg/day dose group in males and 2620 mg/Kg/day dose group in females. Water consumption showed a dose-related increase in the two highest dose groups (190 and 2300 mg/Kg/day and 226 and 2620 mg/Kg/day) and was associated with corresponding urine output. Hematology and chemistry parameters did not show treatment-related effects. An increase in urine output was observed in the two highest dose groups (190 and 2300 mg/Kg/day and 226 and 2620 mg/Kg/day). In the high dose group, significantly increased organ-to-body weight ratios for liver (p<0.01) were seen in males (2300 mg/Kg/day), and in males and females for kidney (p<0.05) (2300 mg/Kg/day for males and 2620 mg/Kg/day for females). The test substance did not adversely affect other parameters. Macroscopic examinations at necropsy revealed an increased incidence of pancreateic nodules and masses in the 2300 mg/kg/day males and 2620 mg/Kg/day females. Histopathology revealed statistically significant test article-related neoplastic effects to the pancreas of high dose males and females. Nodular hyperplasia of the exocrine pancreas was observed in 51 of 78 (p<0.001) high dose (2300 mg/Kg/day) males versus 6 of 78 control group males, and in 10 of 79 (p<0.05) high dose (2620 mg/Kg/day) females versus 1 of 78 control group females. Other non-neoplastic lesions seen in the dosed animals were not related to treatment with FWA-5. Exocrine pancreas adenoma, a benign tumor, was observed in males of the 5,000 and 50,000 ppm dose groups at an incidence of 2/79 and 18/78 (p<0.05), respectively, compared with 0/78 in control. Pancreatic carcinoma, a malignant tumor, was also observed in 2/78 high-dose males and although not statistically significantly increased, the rarity of this tumor and the treatment-related finding of adenomas in this group suggest the carcinomas could be related to the treatment. In females, adenomas in exocrine pancreas occurred only in the high dose (2620 mg/Kg/day) group (2/79) and were within the historical control range; carcinomas were not observed in any of the females. Based on the observations made, the No observed adverse effect level (NOAEL) for the test chemical is considered to be 190 mg/kg/day for males and 226 mg/kg/day for females when they were exposed to the test chemical during the chronic exposure period.

Study 2:

Repeated oral toxicity of test chemical was determined by performing a 80 weeks repeated dose toxicity study using Charles River CD mouse (male and female) at dose levels of 0, 100, 200, 400 or 800 mg/Kg (0.2, 0.4, 0.8 or 1.6% respectively) by oral diet route. The animals were observed for clinical signs, mortality, body weight changes, hematology and were subjected to gross and histopathology. The feeding of Sunset Yellow FCF did not adversely affect the death rate within the groups, the rate of body-weight gain, the organ weights or the haematological findings. The incidence and severity of the histopathological findings were similar in treated and control mice and there was no evidence of an increased incidence of tumours in the mice given Sunset Yellow FCF. Distension of the bladder, frequently associated with hydroureter and hydronephosis was noted. Pus and hard protein deposits were found in the bladders of many of the mice and in a number of cases it was possible to show that an obstruction had occurred in the urethra in the region of the bulbo-urethral complex. After the male mice were put into individual cages, there was a marked reduction in the number showing this syndrome, although isolated cases of urinary retention were seen throughout the study. Of the malignant tumours found, lymphomas and reticulum-cell neoplasms occurred in treated and control mice. In addition a splenic haemangiosarcoma and a mammary carcinoma were found in the female mice fed on the diet containing the lowest level of test chemical (100 mg/Kg). A single tumour of the Harderian gland was found in a female from the group given the 1.6% level. The commonest benign tumours were pulmonary adenomas and benign cysts in the ovary, found with a similar frequency in treated and control mice. The only other tumour occurring in treated mice without a similar finding in the controls was a granulosa-cell tumour of the ovary in a mouse given 800 mg/Kg Sunset Yellow FCF. Thus, on the basis of above results the NOAEL (no observed adverse effect level) for test chemical was considered to be 800 mg/kg diet.

Based on the data available for 2 ,2'-(ethene-1,2-diyldi-4,1-phenylene)bis(1,3-benzoxazole) (CAS no 1533-45-5) does not exhibit repeated dose oral toxicity. Hence the test chemical is not likely to classify as in-vitro gene toxicity as per the criteria mentioned in CLP regulation.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
800 mg/kg bw/day
Study duration:
chronic
Species:
mouse
Quality of whole database:
Data is Kimilsch 2 and from publication

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
repeated dose toxicity: inhalation, other
Data waiving:
other justification
Justification for data waiving:
other:
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

Link to relevant study records
Reference
Endpoint:
repeated dose toxicity: dermal, other
Data waiving:
other justification
Justification for data waiving:
other:
Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Repeated dose oral toxicity:

Data available for target and the read across chemicals was reviewed to determine the repeated dose oral toxicity of 2 ,2'-(ethene-1,2-diyldi-4,1-phenylene)bis(1,3-benzoxazole) (CAS no 1533-45-5). The studies are as mentioned below:

Study 1:

Repeated dose oral toxicity study was performed to determine the toxic nature of the test chemical. The study was performed using male and female Tif:RAIf, SPF strain rats. The test chemical was mixed with feed at dose levels of 0, 500, 5000, or 50,000 ppm (19, 190, and 2300 mg/kg/day in males and 21, 226, and 2620 mg/kg/day in females). The test substance did not affect appearance and behavior and was similar in all groups of test. Mortality was unaffected by the test chemical. 10% decrease in body weight was observed in 2300 mg/Kg/day dose group in males and 2620 mg/Kg/day dose group in females. Increased food consumption was observed in 2300 mg/Kg/day dose group in males and 2620 mg/Kg/day dose group in females. Water consumption showed a dose-related increase in the two highest dose groups (190 and 2300 mg/Kg/day and 226 and 2620 mg/Kg/day) and was associated with corresponding urine output. Hematology and chemistry parameters did not show treatment-related effects. An increase in urine output was observed in the two highest dose groups (190 and 2300 mg/Kg/day and 226 and 2620 mg/Kg/day). In the high dose group, significantly increased organ-to-body weight ratios for liver (p<0.01) were seen in males (2300 mg/Kg/day), and in males and females for kidney (p<0.05) (2300 mg/Kg/day for males and 2620 mg/Kg/day for females). The test substance did not adversely affect other parameters. Macroscopic examinations at necropsy revealed an increased incidence of pancreateic nodules and masses in the 2300 mg/kg/day males and 2620 mg/Kg/day females. Histopathology revealed statistically significant test article-related neoplastic effects to the pancreas of high dose males and females. Nodular hyperplasia of the exocrine pancreas was observed in 51 of 78 (p<0.001) high dose (2300 mg/Kg/day) males versus 6 of 78 control group males, and in 10 of 79 (p<0.05) high dose (2620 mg/Kg/day) females versus 1 of 78 control group females. Other non-neoplastic lesions seen in the dosed animals were not related to treatment with FWA-5. Exocrine pancreas adenoma, a benign tumor, was observed in males of the 5,000 and 50,000 ppm dose groups at an incidence of 2/79 and 18/78 (p<0.05), respectively, compared with 0/78 in control. Pancreatic carcinoma, a malignant tumor, was also observed in 2/78 high-dose males and although not statistically significantly increased, the rarity of this tumor and the treatment-related finding of adenomas in this group suggest the carcinomas could be related to the treatment. In females, adenomas in exocrine pancreas occurred only in the high dose (2620 mg/Kg/day) group (2/79) and were within the historical control range; carcinomas were not observed in any of the females. Based on the observations made, the No observed adverse effect level (NOAEL) for the test chemical is considered to be 190 mg/kg/day for males and 226 mg/kg/day for females when they were exposed to the test chemical during the chronic exposure period.

Study 2:

Repeated oral toxicity of test chemical was determined by performing a 80 weeks repeated dose toxicity study using Charles River CD mouse (male and female) at dose levels of 0, 100, 200, 400 or 800 mg/Kg (0.2, 0.4, 0.8 or 1.6% respectively) by oral diet route. The animals were observed for clinical signs, mortality, body weight changes, hematology and were subjected to gross and histopathology. The feeding of Sunset Yellow FCF did not adversely affect the death rate within the groups, the rate of body-weight gain, the organ weights or the haematological findings. The incidence and severity of the histopathological findings were similar in treated and control mice and there was no evidence of an increased incidence of tumours in the mice given Sunset Yellow FCF. Distension of the bladder, frequently associated with hydroureter and hydronephosis was noted. Pus and hard protein deposits were found in the bladders of many of the mice and in a number of cases it was possible to show that an obstruction had occurred in the urethra in the region of the bulbo-urethral complex. After the male mice were put into individual cages, there was a marked reduction in the number showing this syndrome, although isolated cases of urinary retention were seen throughout the study. Of the malignant tumours found, lymphomas and reticulum-cell neoplasms occurred in treated and control mice. In addition a splenic haemangiosarcoma and a mammary carcinoma were found in the female mice fed on the diet containing the lowest level of test chemical (100 mg/Kg). A single tumour of the Harderian gland was found in a female from the group given the 1.6% level. The commonest benign tumours were pulmonary adenomas and benign cysts in the ovary, found with a similar frequency in treated and control mice. The only other tumour occurring in treated mice without a similar finding in the controls was a granulosa-cell tumour of the ovary in a mouse given 800 mg/Kg Sunset Yellow FCF. Thus, on the basis of above results the NOAEL (no observed adverse effect level) for test chemical was considered to be 800 mg/kg diet.

Based on the data available for 2 ,2'-(ethene-1,2-diyldi-4,1-phenylene)bis(1,3-benzoxazole) (CAS no 1533-45-5) does not exhibit repeated dose oral toxicity. Hence the test chemical is not likely to classify as in-vitro gene toxicity as per the criteria mentioned in CLP regulation.

Repeated dose dermal toxicity:

2,2'-(Vinylenedi-4-phenylene)bis(benzoxazole) has a very low vapor pressure (1.28E-013 mmHg), so the potential for the generation of inhalable vapours is very low. Also the normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be highly unlikely and therefore this end point for repeated inhalation toxicity was considered for waiver.

Repeated dose inhalation toxicity:

The acute dermal toxicity value for 2,2'-(Vinylenedi-4-phenylene)bis(benzoxazole) (as provided in section 7.2.3) is >2000 mg/kg body weight. The substance was also found to be not irritating and not sensitizing to the skin. Based on these considerations, the end point for repeated dermal toxicity is considered as waiver.

Justification for classification or non-classification

Based on the data available for 2 ,2'-(ethene-1,2-diyldi-4,1-phenylene)bis(1,3-benzoxazole) (CAS no 1533-45-5) does not exhibit repeated dose oral, dermal and inahation toxicity. Hence the test chemical is not likely to classify as in-vitro gene toxicity as per the criteria mentioned in CLP regulation.