Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Repeated Dose Oral Toxicity:

Based on the presented data, the target chemical is expected not to be toxic to rats and mice following repeated dose exposure.

Repeated Dose Inhalation Toxicity

A short-term toxicity study need not be conducted as because exposure of humans via inhalation route in production and/or use is not likely based on the provided thorough and rigorous exposure assessment. The estimated vapor pressure of the test chemical was 3.49E-10 mm Hg.Hence, this endpoint can be considered for waiver

Repeated Dose dermal Toxicity

A short-term toxicity study need not be conducted as because exposure of humans via dermal route in production and/or use is not likely based on the provided thorough and rigorous exposure assessment.The acute dermal LD50 of the test chemical can be considered to be >2000 mg/kgbw. Hence, this endpoint was considered for waiver.

Key value for chemical safety assessment

Toxic effect type:
dose-dependent

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:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
secondary literature
Justification for type of information:
Data is from a secondary literature.
Qualifier:
according to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Principles of method if other than guideline:
According to OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
GLP compliance:
not specified
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
No Data Available
Route of administration:
oral: gavage
Vehicle:
other: Homogenized bi-distilled water with 1% CMC
Details on oral exposure:
No Data Available
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No Data Available
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Daily
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Control Group
Dose / conc.:
100 mg/kg bw/day (nominal)
Remarks:
Low Dose Group
Dose / conc.:
300 mg/kg bw/day (nominal)
Remarks:
Mid Dose Group
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
High Dose Group
No. of animals per sex per dose:
20 animals (10 males & 10 females) / dose level
Control animals:
yes, concurrent vehicle
Details on study design:
No Data Available
Positive control:
No Data Available
Observations and examinations performed and frequency:
Animals were observed twice daily for mortality/morbidity and once daily for clinical abnormalities. Individual animal weights were recorded weekly. Body weight and food consumption were recorded weekly. Ophthalmologic evaluations on control and high-dose animals were performed at the end of the study. Haematology, clinical
chemistry and urinalysis evaluations were performed once during week 13.
Sacrifice and pathology:
At the end of the treatment period, all animals were killed and grossly examined. Selected organs were weighted. All animals were submitted to a complete macroscopic examination.
Other examinations:
No Data Available
Statistics:
No Data Available
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Clinical signs were restricted to discolorations of faeces and the mucosal surface of the stomach and/or intestines at ≥100 mg/kg and discolorations of the tail and paws at 1000 mg/kg.
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No significant changes in body weight or food intake were observed.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No significant changes in body weight or food intake were observed.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
no effects observed
Description (incidence and severity):
Ophthalmological findings were unremarkable.
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Male rats treated at 1000 mg/kg showed decreased fibrinogen levels.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
Male rats treated at 1000 mg/kg showed slightly increased beta-globulin levels compared to the control group. Female rats treated at 1000 mg/kg showed decreased bilirubin levels and increased phospholipid levels compared to the control group. The observed biochemical changes lacked histopathologic correlates.
Urinalysis findings:
effects observed, non-treatment-related
Description (incidence and severity):
Decreased uric acid levels were observed at 300 mg/kg (females) and at 1000 mg/kg (both genders) compared to the control data.
Behaviour (functional findings):
effects observed, non-treatment-related
Description (incidence and severity):
Male rats treated at 1000 mg/kg showed slightly increased locomotor activity.
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
Clinical signs were restricted to discolorations of faeces and the mucosal surface of the stomach and/or intestines at ≥100 mg/kg and discolorations of the tail and paws at 1000 mg/kg. No significant changes in body weight or food intake were observed. Ophthalmological findings were unremarkable. Decreased uric acid levels were observed at 300 mg/kg (females) and at 1000 mg/kg (both genders) compared to the control data. Male rats treated at 1000 mg/kg showed slightly increased locomotor activity, decreased fibrinogen levels, and increased beta-globulin levels compared to the control group. Female rats treated at 1000 mg/kg showed decreased bilirubin levels and increased phospholipid levels compared to the control group. The observed biochemical changes lacked histopathologic correlates. Therefore, the biochemical changes were considered to be adaptive changes with no toxicological significance
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
behaviour (functional findings)
body weight and weight gain
clinical biochemistry
clinical signs
food consumption and compound intake
gross pathology
haematology
histopathology: non-neoplastic
mortality
ophthalmological examination
organ weights and organ / body weight ratios
urinalysis
Remarks on result:
not determinable due to absence of adverse toxic effects
Critical effects observed:
no
Conclusions:
Based on all the available data, it was concluded that the study-derived NOAEL was 1000 mg/kg bw/day.
Executive summary:

The read-across analogue hydrogen 3,6-bis(diethylamino)-9-(2,4disulphonatophenyl)xanthylium, sodium salt[CAS: 3520-42-1; EC: 222-529-8]has been tested for repeated dose toxicity according to OECD 408 (1998) as reported by the Scientific Committee on Consumer Products (SCCP/1115/07). The chemical was given by oral gavage to 10 rats per sex per dose level at 0 (vehicle), 100, 300 and 1000 mg/kg bw/day for a total of 13 weeks.No mortality was observed during the treatment period. Clinical signs were restricted to discolorations of faeces and the mucosal surface of the stomach and/or intestines at ≥100 mg/kg and discolorations of the tail and paws at 1000 mg/kg. No significant changes in body weight or food intake were observed. Ophthalmological findings were unremarkable. Decreased uric acid levels were observed at 300 mg/kg (females) and at 1000 mg/kg (both genders) compared to the control data. Male rats treated at 1000 mg/kg showed slightly increased locomotor activity, decreased fibrinogen levels, and increased beta-globulin levels compared to the control group. Female rats treated at 1000 mg/kg showed decreased bilirubin levels and increased phospholipid levels compared to the control group. The observed biochemical changes lacked histopathologic correlates. Therefore, the biochemical changes were considered to be adaptive changes with no toxicological significance. The study-derived NOAEL was 1000 mg/kg bw/day. The study was performed according to GLP.

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Justification for type of information:
Data is from a NTP report.
Qualifier:
according to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Principles of method if other than guideline:
According to OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
GLP compliance:
not specified
Species:
rat
Strain:
Fischer 344
Remarks:
Fischer 344/N
Sex:
male/female
Details on test animals or test system and environmental conditions:
No data Available
Route of administration:
oral: feed
Vehicle:
other: diet
Details on oral exposure:
The chemical was given by diet to 10 rats per sex per dose level at 0, 125, 250, 500, 1000 and 2000 ppm for 13 weeks via feed route.
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No data available
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Daily
Remarks:
0, 125, 250, 500, 1000 and 2000 ppm
No. of animals per sex per dose:
10 rats per sex per dose
Control animals:
yes, plain diet
Details on study design:
No data available
Positive control:
No data available
Observations and examinations performed and frequency:
The animals were observed twice per day for mortality, morbidity and clinical signs of toxicity. Body weight and food intake were recorded once per week. All animals found dead and those that survived treatment were necropsied.
Sacrifice and pathology:
All animals found dead and those that survived treatment were necropsied.The following organs, whenever possible, were examined grossly and histologically: adrenals, brain colon, oesophagus, femur including marrow heart, kidneys, liver lungs and bronchi, mammary gland, mandibular and mesenteric lymph nodes, pancreas parathyroid glands, pituitary gland, salivary glands, seminal vesicles, prostate, testes, ovaries, uterus, skin, small intestines, spleen, stomach, thigh muscle, thymus, thyroid gland trachea, and the urinary bladder.
Other examinations:
No Data Available
Statistics:
The probability of survival was estimated by the product-limit procedure of Kaplan and Meier (1958) and is presented in the form of graphs. Animals were censored from the survival analyses at the time they were found to be missing or dead from other than natural causes; animals dying from natural causes were not censored. Statistical analyses for a possible dose-related effect on survival used the method of Cox (1972) for testing two groups for equality and Tarone’s (1975) life table test for a dose-related trend. When significant survival differ- ences were detected, additional analyses using these procedures were carried out to determine the time point at which significant differences in the survival curves were first detected. All re- ported P values for the survival analysis are two-sided.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Clinical findings were limited to discoloured faecal matter at ≥125 ppm.
Mortality:
no mortality observed
Description (incidence):
All animals survived to planned death.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
The mean terminal body weights of rats treated at 500, 1000 and 2000 ppm were 12, 13, and 32% lower than the control data for males and 4, 8, and 20% lower for females.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
Food intake was somewhat lower at 2000 ppm.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
Bone marrow atrophy was observed at increased incidence and severity at ≥500 ppm.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Bone marrow atrophy was observed at increased incidence and severity at ≥500 ppm. This effect was of minimal severity in 5/10 males and 4/10 females at 500 ppm, of mild severity in 10/10 males and 8/10 females at 1000 ppm, and of moderate severity in 10/10 males and 9/9 females at 2000 ppm.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
Clinical findings were limited to discoloured faecal matter at ≥125 ppm. The mean terminal body weights of rats treated at 500, 1000 and 2000 ppm were 12, 13, and 32% lower than the control data for males and 4, 8, and 20% lower for females. Food intake was somewhat lower at 2000 ppm. Bone marrow atrophy was observed at increased incidence and severity at ≥500 ppm. This effect was of minimal severity in 5/10 males and 4/10 females at 500 ppm, of mild severity in 10/10 males and 8/10 females at 1000 ppm, and of moderate severity in 10/10 males and 9/9 females at 2000 ppm.
Dose descriptor:
NOAEL
Effect level:
250 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
clinical signs
food consumption and compound intake
gross pathology
histopathology: non-neoplastic
mortality
Remarks on result:
not determinable due to absence of adverse toxic effects
Critical effects observed:
no
Conclusions:
Based on all the observations and results, it was concluded that the NOAEL for the test chemical was observed to be 250 ppm.
Executive summary:

The read-across analogue2-[6-(ethylamino)-3-(ethylimino)2,7-dimethyl-3-xanthen-9-yl]benzoic acid ethyl ester, monohydrochloride[CAS: 989-38-8; EC: 213-584-9]has been tested for repeated dose toxicity by the National Toxicology Program (TR 364). The chemical was given by diet to 10 rats per sex per dose level at 0, 125, 250, 500, 1000 and 2000 ppm for 13 weeks. The animals were observed twice per day for mortality, morbidity and clinical signs of toxicity. Body weight and food intake were recorded once per week. All animals found dead and those that survived treatment were necropsied.The following organs, whenever possible, were examined grossly and histologically:adrenals, braincolon, oesophagus, femurincluding marrow heart, kidneys, liverlungs and bronchi, mammarygland, mandibular and mesenteric lymph nodes, pancreasparathyroid glands, pituitarygland, salivary glands, seminalvesicles,prostate,testes,ovaries,uterus, skin, small intestines, spleen, stomach, thighmuscle, thymus, thyroid glandtrachea, andtheurinary bladder. All animals survived to planned death. Clinical findings were limited to discoloured faecal matter at ≥125 ppm. The mean terminal body weights of rats treated at 500, 1000 and 2000 ppm were 12, 13, and 32% lower than the control data for males and 4, 8, and 20% lower for females. Food intake was somewhat lower at 2000 ppm. Bone marrow atrophy was observed at increased incidence and severity at ≥500 ppm. This effect was of minimal severity in 5/10 males and 4/10 females at 500 ppm, of mild severity in 10/10 males and 8/10 females at 1000 ppm, and of moderate severity in 10/10 males and 9/9 females at 2000 ppm. NOAEL for the test chemical in rats was considered at 250 ppm. The study was performed according to GLP.

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Justification for type of information:
Data is from a NTP report.
Qualifier:
according to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Principles of method if other than guideline:
According to OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
GLP compliance:
not specified
Species:
rat
Strain:
Fischer 344
Remarks:
Fischer 344/N
Sex:
male/female
Details on test animals or test system and environmental conditions:
No data Available
Route of administration:
oral: feed
Vehicle:
other: diet
Details on oral exposure:
The chemical was given by diet to 10 rats per sex per dose level at 0, 125, 250, 500, 1000 and 2000 ppm for 13 weeks via feed route.
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No data available
Duration of treatment / exposure:
103 weeks
Frequency of treatment:
Daily
Remarks:
0, 125 and 250 ppm
No. of animals per sex per dose:
10 rats per sex per dose
Control animals:
yes, plain diet
Details on study design:
No data available
Positive control:
No data available
Observations and examinations performed and frequency:
The animals were observed twice per day for mortality, morbidity and clinical signs of toxicity. Body weight and food intake were recorded once per week. All animals found dead and those that survived treatment were necropsied.
Sacrifice and pathology:
All animals found dead and those that survived treatment were necropsied.The following organs, whenever possible, were examined grossly and histologically: adrenals, brain colon, oesophagus, femur including marrow heart, kidneys, liver lungs and bronchi, mammary gland, mandibular and mesenteric lymph nodes, pancreas parathyroid glands, pituitary gland, salivary glands, seminal vesicles, prostate, testes, ovaries, uterus, skin, small intestines, spleen, stomach, thigh muscle, thymus, thyroid gland trachea, and the urinary bladder.
Other examinations:
No Data Available
Statistics:
The probability of survival was estimated by the product-limit procedure of Kaplan and Meier (1958) and is presented in the form of graphs. Animals were censored from the survival analyses at the time they were found to be missing or dead from other than natural causes; animals dying from natural causes were not censored. Statistical analyses for a possible dose-related effect on survival used the method of Cox (1972) for testing two groups for equality and Tarone’s (1975) life table test for a dose-related trend. When significant survival differ- ences were detected, additional analyses using these procedures were carried out to determine the time point at which significant differences in the survival curves were first detected. All re- ported P values for the survival analysis are two-sided.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Clinical signs were limited to discoloured fur and skin at ≥125 ppm.
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Male rats treated at 125 and 250 ppm weighed slightly more (about 3-7%) than the control group from week 77 to the end of the study period. Female rats treated at 125 and 250 ppm weighed about the same as the control group throughout the entire study period.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No significant effects on food intake were observed.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
No treatment-related gross pathological effects were observed.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Neoplastic findings were limited to increased incidences of keratoacanthomas of the skin (male rats treated at 250 ppm) and pheochromocytomas or malignant pheochromocytomas (female rats treated at 250 ppm). Neither of the two neoplasms were conclusively attributed to the test chemical. No non-neoplastic lesions attributed to the test chemical were observed.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
No treatment-related effects on mortality were observed. Clinical signs were limited to discoloured fur and skin at ≥125 ppm. Male rats treated at 125 and 250 ppm weighed slightly more (about 3-7%) than the control group from week 77 to the end of the study period. Female rats treated at 125 and 250 ppm weighed about the same as the control group throughout the entire study period. No significant effects on food intake were observed. No treatment-related gross pathological effects were observed. Neoplastic findings were limited to increased incidences of keratoacanthomas of the skin (male rats treated at 250 ppm) and pheochromocytomas or malignant pheochromocytomas (female rats treated at 250 ppm). Neither of the two neoplasms were conclusively attributed to the test chemical. No non-neoplastic lesions attributed to the test chemical were observed. The authors of the study acknowledge that the rats could potentially have tolerated a higher dose than 250 ppm. The dose of 250 ppm corresponded to 10 (males) and 12 mg/kg bw/day (females). The study was performed according to GLP.
Dose descriptor:
NOAEL
Effect level:
250 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
clinical signs
food consumption and compound intake
gross pathology
histopathology: non-neoplastic
mortality
Remarks on result:
not determinable due to absence of adverse toxic effects
Critical effects observed:
no
Conclusions:
Based on all the observations and results, it was concluded that the NOAEL for the test chemical was observed to be 250 ppm.
Executive summary:

The read-across analogue2 -[6 -(ethylamino)-3 -(ethylimino)2,7 -dimethyl-3 -xanthen-9-yl]benzoic acid ethyl ester, monohydrochloride[CAS: 989-38-8; EC: 213-584-9]has been tested for repeated dose toxicity by the National Toxicology Program (TR 364). The chemical was given by diet to 50 rats per sex per dose level at 0, 125 and 250 ppm for 103 weeks. The animals were observed twice per day for mortality, morbidity and clinical signs of toxicity. Body weight and food intake were recorded once per week during the first 13 weeks of treatment and monthly thereafter. All animals found dead and those that survived treatment were necropsied.Except for the aorta, vagina, coagulation glands and peripheral nerve, the gross and histopathological examinations were performed as per the latest OECD 408.No treatment-related effects on mortality were observed. Clinical signs were limited to discoloured fur and skin at ≥125 ppm. Male rats treated at 125 and 250 ppm weighed slightly more (about 3-7%) than the control group from week 77 to the end of the study period. Female rats treated at 125 and 250 ppm weighed about the same as the control group throughout the entire study period. No significant effects on food intake were observed. No treatment-related gross pathological effects were observed. Neoplastic findings were limited to increased incidences of keratoacanthomas of the skin (male rats treated at 250 ppm) andpheochromocytomas or malignant pheochromocytomas(female rats treated at 250 ppm). Neither of the two neoplasms were conclusively attributed to the test chemical. No non-neoplastic lesions attributed to the test chemical were observed. The authors of the study acknowledge that the rats could potentially have tolerated a higher dose than 250 ppm. The dose of 250 ppm corresponded to 10 (males) and 12 mg/kg bw/day (females).The study was performed according to GLP.

Endpoint:
chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from a journal
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 453 (Combined Chronic Toxicity / Carcinogenicity Studies)
Principles of method if other than guideline:
Equivalent or similar to OECD Guideline 453 (Combined Chronic Toxicity / Carcinogenicity Studies)
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
No Data Available
Route of administration:
oral: feed
Vehicle:
other: Diet
Details on oral exposure:
No data available
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No Data Available
Duration of treatment / exposure:
2 years
Frequency of treatment:
Daily
Remarks:
0, 4.4, 45 and 450.7 mg/kg bw/day (males) and 0, 6.2, 62.5 and 630.1 mg/kg bw/day (females).
No. of animals per sex per dose:
48 rats per sex per dose level
Control animals:
yes, plain diet
Details on study design:
No Data Available
Positive control:
No Data Available
Observations and examinations performed and frequency:
The animals were observed once per day for mortality, morbidity, and clinical signs of toxicity. Body weight, food intake, and water consumption were recorded initially (body weight only), once per two weeks during the first year, and monthly during the second year. Blood samples were collected from 12 rats per sex per dose level at 14, 26 and 54 weeks of study, and from all surviving rats at 24 months of study. Blood parameters included total red blood cell and white blood cell counts, differential white blood cell counts, haemoglobin concentrations, packed cell volume, and reticulocyte count. Urine samples were collected from ≥10 rats per sex per dose level at 2 months intervals until 24 months of study, at which time urine samples were collected from all surviving rats. Urine samples collected over a 6-hour period were examined for volume, specific gravity, pH, glucose, blood, bilirubin, ketones, and protein. Urine samples collected over a 2-hour period following an oral water load of 25 ml/kg were examined for volume, specific gravity and cell content. Urine samples collected over a 6-hour period following an oral water load of 25 ml/kg were examined for volume and specific gravity.
Sacrifice and pathology:
All animals found dead and those that survived treatment were necropsied. The major organs were weighed, and the following organs were examined microscopically for neoplastic and non-neoplastic lesions: adrenal, aorta, bladder, brain, caecum, colon, duodenum, eye, Harderian gland, heart, ileum, kidney, liver, lung, lymph node, mammary gland, muscle, nerve, oesophagus, ovary, pancreas, pituitary, prostate, rectum, salivary gland, seminal vesicles, spinal cord, spleen, stomach, testis, thymus, thyroid, trachea, uterus, and vagina.
Other examinations:
No Data Available
Statistics:
No Data Available
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Male rats treated at 450.7 mg/kg bw/day showed an increased incidence of pneumonia compared to the control group. No treatment-related effects on clinical signs.
Mortality:
no mortality observed
Description (incidence):
No treatment-related effects on mortality were observed.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No treatment-related effects on the body weight test animals were observed.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No treatment-related effects on the feed consumption on the test animals were observed.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
effects observed, non-treatment-related
Description (incidence and severity):
Reduced water intake during the first 9 months of study was observed at 450.7 (males) and 630.1 mg/kg bw/day (females).
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Description (incidence and severity):
No treatment-related effects on the haematological findings on the test animals were observed.
Clinical biochemistry findings:
not specified
Urinalysis findings:
effects observed, non-treatment-related
Description (incidence and severity):
Except for increased number of cells excreted in the 2-hour urine samples, most notably at mid- and high dose levels, no significant effects in urinalysis were observed.
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
No treatment-related effects on mortality were observed. Male rats treated at 450.7 mg/kg bw/day showed an increased incidence of pneumonia compared to the control group. This effect was not attributed to the test chemical in the report. No treatment-related effects on clinical signs, body weight, food intake, or haematology were observed. Reduced water intake during the first 9 months of study was observed at 450.7 (males) and 630.1 mg/kg bw/day (females). Except for increased number of cells excreted in the 2-hour urine samples, most notably at mid- and high dose levels, no significant effects in urinalysis were observed. No histopathological effects of toxicological significance were observed.
Dose descriptor:
NOAEL
Effect level:
4.4 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
behaviour (functional findings)
body weight and weight gain
clinical signs
food consumption and compound intake
gross pathology
haematology
histopathology: non-neoplastic
mortality
urinalysis
water consumption and compound intake
Remarks on result:
not determinable due to absence of adverse toxic effects
Critical effects observed:
no
Conclusions:
Based on the available data the panel-derived NOAELs were 4.4 (males) and 6.2 mg/kg bw/day (females).
Executive summary:

The read-across analogueTetrasodium hexacyanoferrate[CAS 13601-19-9; EC: 237-081-9]has been tested for repeated dose toxicity as reported by the European Food Safety Authority (2018). The chemical was given by diet to 48 rats per sex per dose level for a total of two years. Dose levels were equal to about 0, 4.4, 45 and 450.7 mg/kg bw/day (males) and 0, 6.2, 62.5 and 630.1 mg/kg bw/day (females). The animals were observed once per day for mortality, morbidity, and clinical signs of toxicity. Body weight, food intake, and water consumption were recorded initially (body weight only), once per two weeks during the first year, and monthly during the second year. Blood samples were collected from 12 rats per sex per dose level at 14, 26 and 54 weeks of study, and from all surviving rats at 24 months of study. Blood parameters included total red blood cell and white blood cell counts, differential white blood cell counts, haemoglobin concentrations, packed cell volume, and reticulocyte count. Urine samples were collected from ≥10 rats per sex per dose level at 2 months intervals until 24 months of study, at which time urine samples were collected from all surviving rats. Urine samples collected over a 6-hour period were examined for volume, specific gravity, pH, glucose, blood, bilirubin, ketones, and protein. Urine samples collected over a 2-hour period following an oral water load of 25 ml/kg were examined for volume, specific gravity and cell content. Urine samples collected over a 6-hour period following an oral water load of 25 ml/kg were examined for volume and specific gravity.All animals found dead and those that survived treatment were necropsied. The major organs were weighed, and the following organs were examined microscopically for neoplastic and non-neoplastic lesions:adrenal, aorta, bladder, brain, caecum, colon, duodenum, eye, Harderian gland,heart, ileum, kidney, liver, lung, lymph node, mammarygland, muscle, nerve, oesophagus, ovary, pancreas,pituitary, prostate, rectum, salivarygland, seminal vesicles, spinal cord, spleen, stomach, testis, thymus, thyroid,trachea, uterus,and vagina. No treatment-related effects on mortality were observed. Male rats treated at 450.7 mg/kg bw/day showed an increased incidence of pneumonia compared to the control group. This effect was not attributed to the test chemical in the report. No treatment-related effects on clinical signs, body weight, food intake, or haematology were observed. Reduced water intake during the first 9 months of study was observed at 450.7 (males) and 630.1 mg/kg bw/day (females). Except for increased number of cells excreted in the 2-hour urine samples, most notably at mid- and high dose levels, no significant effects in urinalysis were observed. No histopathological effects of toxicological significance were observed. The panel-derived NOAELs were 4.4 (males) and 6.2 mg/kg bw/day (females). The study was performed pre-GLP.

Endpoint:
chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Justification for type of information:
Data is from a NTP report.
Qualifier:
according to guideline
Guideline:
OECD Guideline 453 (Combined Chronic Toxicity / Carcinogenicity Studies)
Principles of method if other than guideline:
According to OECD Guideline 453 (Combined Chronic Toxicity / Carcinogenicity Studies)
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
Fischer 344
Sex:
male/female
Details on test animals or test system and environmental conditions:
No Data Available
Route of administration:
oral: feed
Vehicle:
other: Diet
Details on oral exposure:
No data available
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No Data Available
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Daily
Remarks:
0, 500, 1000, 2000, 4000 and 8000 ppm
No. of animals per sex per dose:
10 animals per sex per dose
Control animals:
yes, plain diet
Details on study design:
No Data Available
Positive control:
No Data Available
Observations and examinations performed and frequency:
The animals were observed twice per day for mortality and morbidity. Clinical observations were recorded once per week. Body weights were recorded prior to treatment, on day 1 of treatment, and weekly thereafter. Food intake was recorded once per week. Blood samples were collected from all surviving rats at the end of the study period. Haematology parameters included haematocrit, haemoglobin, erythrocytes, reticulocytes, nucleated erythrocytes, mean cell volume, mean cell hemoglobin, mean cell hemoglobin concentration, platelets, white blood cell counts, and differential white blood cell counts. Clinical chemistry parameters included ALT, ALP, 5'-nucleotidase, SDH, bile salts, total protein, albumin, creatinine, and urea nitrogen. Urine samples were collected from all surviving rats at the end of the study period. Urine parameters included creatinine, glucose, protein, AST, NAG, urine volume, and specific gravity.
Sacrifice and pathology:
Each animal was given a complete necropsy. Body weights of the following organs were recorded: liver, thymus, right kidney, right testis, heart, lungs, and brain. The following organs were examined grossly and histologically at 0 and ≤8000 ppm: adrenal glands, brain (three sections), oesophagus, eyes (if grossly abnormal), femur with marrow, gross lesions, heart, intestines (large: cecum, colon, rectum; small: duodenum, jejunum, ileum), kidneys, liver, lung/mainstem bronchi, lymph nodes (mandibular, mesenteric), mammary gland, nasal cavity and turbinates (three sections), ovaries, pancreas, parathyroid glands, pharynx (if grossly abnormal), pituitary gland, preputial or clitoral glands, prostate gland, salivary glands, spinal cord/sciatic nerve (if neurologic signs were present), spleen, stomach (forestomach and glandular stomach), testes (with epididymis), thymus, thyroid gland, trachea, urinary bladder, and uterus. In addition to the dosed animals above, supplemental animals (10 rats/sex/dose) treated at 0, 500, 1000, 2000, 4000 and 8000 ppm were included for interim evaluations (on days 5 and 21) of haematology, clinical chemistry, and urinalysis.
The supplemental animals treated at 0, 500, 1000, 2000 and 4000 ppm were examined for sperm morphology and vaginal cytology.
Other examinations:
No Data Available
Statistics:
No Data Available
Clinical signs:
no effects observed
Description (incidence and severity):
No clinical signs of toxicity attributed to the test chemical were observed.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
All animals survived to planned death, apart from one female rat (at 1000 ppm) that was killed by accident.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
Average terminal body weights relative to the controls were 92, 99, 98, 93 and 76% (males) and 101, 103, 101, 97, and 93% (females) at 500, 1000, 2000, 4000 and 8000 ppm, respectively.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No remarkable effects on food intake were observed at 500, 1000, 2000 or 4000 ppm. Food intake by male and female rats treated at 8000 ppm were slightly below that of the controls.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
Treatment-related effects on haematology for the most part, restricted to the 4000 and 8000 ppm groups.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
Treatment-related effects on clinical chemistry were, for the most part, restricted to the 4000 and 8000 ppm groups.
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
Treatment-related effects on urinalysis were, for the most part, restricted to the 4000 and 8000 ppm groups.
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
No significant changes in absolute organ weights were observed up to 4000 ppm. Relative organ weights for dosed animals were similar to those of the controls or increased with decreasing mean body weight at 4000 and 8000 ppm.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
Gross lesions attributed to the test chemical were limited to forestomach lesions at 2000 ppm and above.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
The histopathological examination revealed an increased incidence of hyperplasia and hyperkeratosis of the forestomach at 2000 ppm and above; an increased incidence of inflammation of the liver at 2000 ppm and above; and an increased incidence in the number (and size) of protein droplets in the epithelial cytoplasm and the lumen of the proximal convoluted tubules at 2000 ppm and above. The mean severity of each histopathological finding at 2000 ppm never exceeded 1.6 according to the grading scale in the study (1=minimal, 2=mild, 3=moderate, and 4=marked). Histopathological findings at 1000 ppm were limited to one incidence of cytoplasmic alteration (kidneys) which was described as minimal in nature. No significant effects on sperm morphology or vaginal cytology were observed.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
All animals survived to planned death, apart from one female rat (at 1000 ppm) that was killed by accident. No clinical signs of toxicity attributed to the test chemical were observed. Average terminal body weights relative to the controls were 92, 99, 98, 93 and 76% (males) and 101, 103, 101, 97, and 93% (females) at 500, 1000, 2000, 4000 and 8000 ppm, respectively. No remarkable effects on food intake were observed at 500, 1000, 2000 or 4000 ppm. Food intake by male and female rats treated at 8000 ppm were slightly below that of the controls. Treatment-related effects on haematology, clinical chemistry and urinalysis were, for the most part, restricted to the 4000 and 8000 ppm groups. The observed effects were indicative of hepatocellular damage, of compensatory responses to anaemia by the bone marrow, of and renal tubule epithelial damage. No significant changes in absolute organ weights were observed up to 4000 ppm. Relative organ weights for dosed animals were similar to those of the controls or increased with decreasing mean body weight at 4000 and 8000 ppm. Gross lesions attributed to the test chemical were limited to forestomach lesions at 2000 ppm and above. The histopathological examination revealed an increased incidence of hyperplasia and hyperkeratosis of the forestomach at 2000 ppm and above; an increased incidence of inflammation of the liver at 2000 ppm and above; and an increased incidence in the number (and size) of protein droplets in the epithelial cytoplasm and the lumen of the proximal convoluted tubules at 2000 ppm and above. The mean severity of each histopathological finding at 2000 ppm never exceeded 1.6 according to the grading scale in the study (1=minimal, 2=mild, 3=moderate, and 4=marked). Histopathological findings at 1000 ppm were limited to one incidence of cytoplasmic alteration (kidneys) which was described as minimal in nature. No significant effects on sperm morphology or vaginal cytology were observed.
Dose descriptor:
NOAEL
Effect level:
1 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
clinical biochemistry
clinical signs
food consumption and compound intake
gross pathology
haematology
histopathology: non-neoplastic
mortality
organ weights and organ / body weight ratios
urinalysis
Remarks on result:
not determinable due to absence of adverse toxic effects
Critical effects observed:
no
Conclusions:
Based on all the observations and results, NOAEL for the test chemical was considered at 1000 ppm. The dose of 1000 ppm corresponded to 64 (males) and 68 mg/kg bw/day (females).
Executive summary:

The read-across analogue cupric sulfate pentahydrate [CAS: 7758-99-8; EC: 616-477-9]has been tested for repeated dose toxicity by the National Toxicology Program (1993). The chemical was given by diet to 10 rats per sex per dose level at 0, 500, 1000, 2000, 4000 and 8000 ppm. The animals were observed twice per day for mortality and morbidity. Clinical observations were recorded once per week. Body weights were recorded prior to treatment, on day 1 of treatment, and weekly thereafter. Food intake was recorded once per week. Blood samples were collected from all surviving rats at the end of the study period. Haematology parameters included haematocrit, haemoglobin, erythrocytes, reticulocytes, nucleated erythrocytes, mean cell volume, mean cell hemoglobin, mean cell hemoglobin concentration, platelets, white blood cell counts, and differential white blood cell counts. Clinical chemistry parameters includedALT, ALP, 5'-nucleotidase, SDH, bile salts, total protein, albumin, creatinine, and urea nitrogen. Urine samples were collected from all surviving rats at the end of the study period. Urine parameters included creatinine, glucose, protein, AST, NAG, urine volume, and specific gravity. Each animal was given a complete necropsy. Body weights of the following organs were recorded: liver, thymus, right kidney, right testis, heart, lungs, and brain. The following organs were examined grossly and histologically at 0 and ≤8000 ppm: adrenal glands, brain (three sections), oesophagus, eyes (if grossly abnormal), femur with marrow, gross lesions, heart, intestines (large: cecum, colon, rectum; small: duodenum, jejunum, ileum), kidneys, liver, lung/mainstem bronchi, lymph nodes (mandibular, mesenteric), mammary gland, nasal cavity and turbinates (three sections), ovaries, pancreas, parathyroid glands, pharynx (if grossly abnormal), pituitary gland, preputial or clitoral glands, prostate gland, salivary glands, spinal cord/sciatic nerve (if neurologic signs were present), spleen, stomach (forestomach and glandular stomach), testes (with epididymis), thymus, thyroid gland, trachea, urinary bladder, and uterus. In addition to the dosed animals above, supplemental animals (10 rats/sex/dose) treated at 0, 500, 1000, 2000, 4000 and 8000 ppm were included for interim evaluations (on days 5 and 21) of haematology, clinical chemistry, and urinalysis.

The supplemental animals treated at 0, 500, 1000, 2000 and 4000 ppm were examined for sperm morphology and vaginal cytology. All animals survived to planned death, apart from one female rat (at 1000 ppm) that was killed by accident. No clinical signs of toxicity attributed to the test chemical were observed. Average terminal body weights relative to the controls were 92, 99, 98, 93 and 76% (males) and 101, 103, 101, 97, and 93% (females) at 500, 1000, 2000, 4000 and 8000 ppm, respectively. No remarkable effects on food intake were observed at 500, 1000, 2000 or 4000 ppm. Food intake by male and female rats treated at 8000 ppm were slightly below that of the controls. Treatment-related effects on haematology, clinical chemistry and urinalysis were, for the most part, restricted to the 4000 and 8000 ppm groups. The observed effects were indicative of hepatocellular damage, of compensatory responses to anaemia by the bone marrow, of and renal tubule epithelial damage. No significant changes in absolute organ weights were observed up to 4000 ppm. Relative organ weights for dosed animals were similar to those of the controls or increased with decreasing mean body weight at 4000 and 8000 ppm. Gross lesions attributed to the test chemical were limited to forestomach lesions at 2000 ppm and above. The histopathological examination revealed an increased incidence of hyperplasia and hyperkeratosis of the forestomach at 2000 ppm and above; an increased incidence of inflammation of the liver at 2000 ppm and above; and an increased incidence in the number (and size) of protein droplets in the epithelial cytoplasm and the lumen of the proximal convoluted tubules at 2000 ppm and above. The mean severity of each histopathological finding at 2000 ppm never exceeded 1.6 according to the grading scale in the study (1=minimal, 2=mild, 3=moderate, and 4=marked). Histopathological findings at 1000 ppm were limited to one incidence of cytoplasmic alteration (kidneys) which was described as minimal in nature. No significant effects on sperm morphology or vaginal cytology were observed. NOAEL for the test chemical was considered at 1000 ppm. The dose of 1000 ppm corresponded to 64 (males) and 68 mg/kg bw/day (females). The study was performed according to GLP.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
Klimisch Rating 2

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
Quality of whole database:
waiver

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
Quality of whole database:
waiver

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Repeated dose Oral toxicity:    

Available data for read-across chemicals were reviewed to predict repeated dose toxicity of the target chemical. The studies are as mentioned below.

Study 1

The read-across analoguehydrogen 3,6-bis(diethylamino)-9-(2,4disulphonatophenyl)xanthylium, sodium salt[CAS: 3520-42-1; EC: 222-529-8]has been tested for repeated dose toxicity according to OECD 408 (1998) as reported by the Scientific Committee on Consumer Products (SCCP/1115/07). The chemical was given by oral gavage to 10 rats per sex per dose level at 0 (vehicle), 100, 300 and 1000 mg/kg bw/day for a total of 13 weeks.No mortality was observed during the treatment period. Clinical signs were restricted to discolorations of faeces and the mucosal surface of the stomach and/or intestines at ≥100 mg/kg and discolorations of the tail and paws at 1000 mg/kg. No significant changes in body weight or food intake were observed. Ophthalmological findings were unremarkable. Decreased uric acid levels were observed at 300 mg/kg (females) and at 1000 mg/kg (both genders) compared to the control data. Male rats treated at 1000 mg/kg showed slightly increased locomotor activity, decreased fibrinogen levels, and increased beta-globulin levels compared to the control group. Female rats treated at 1000 mg/kg showed decreased bilirubin levels and increased phospholipid levels compared to the control group. The observed biochemical changes lacked histopathologic correlates. Therefore, the biochemical changes were considered to be adaptive changes with no toxicological significance. The study-derived NOAEL was 1000 mg/kg bw/day. The study was performed according to GLP.

 

Study 2

The read-across analogue2-[6-(ethylamino)-3-(ethylimino)2,7-dimethyl-3-xanthen-9-yl]benzoic acid ethyl ester, monohydrochloride[CAS: 989-38-8; EC: 213-584-9]has been tested for repeated dose toxicity by the National Toxicology Program (TR 364). The chemical was given by diet to 10 rats per sex per dose level at 0, 125, 250, 500, 1000 and 2000 ppm for 13 weeks. The animals were observed twice per day for mortality, morbidity and clinical signs of toxicity. Body weight and food intake were recorded once per week. All animals found dead and those that survived treatment were necropsied.The following organs, whenever possible, were examined grossly and histologically:adrenals, braincolon, oesophagus, femurincluding marrow heart, kidneys, liverlungs and bronchi, mammarygland, mandibular and mesenteric lymph nodes, pancreasparathyroid glands, pituitarygland, salivary glands, seminalvesicles,prostate,testes,ovaries,uterus, skin, small intestines, spleen, stomach, thighmuscle, thymus, thyroid glandtrachea, andtheurinary bladder. All animals survived to planned death. Clinical findings were limited to discoloured faecal matter at ≥125 ppm. The mean terminal body weights of rats treated at 500, 1000 and 2000 ppm were 12, 13, and 32% lower than the control data for males and 4, 8, and 20% lower for females. Food intake was somewhat lower at 2000 ppm. Bone marrow atrophy was observed at increased incidence and severity at ≥500 ppm. This effect was of minimal severity in 5/10 males and 4/10 females at 500 ppm, of mild severity in 10/10 males and 8/10 females at 1000 ppm, and of moderate severity in 10/10 males and 9/9 females at 2000 ppm. NOAEL for the test chemical in rats was considered at 250 ppm. The study was performed according to GLP.

 

Study 3

The read-across analogue2-[6-(ethylamino)-3-(ethylimino)2,7-dimethyl-3-xanthen-9-yl]benzoic acid ethyl ester, monohydrochloride[CAS: 989-38-8; EC: 213-584-9]has been tested for repeated dose toxicity by the National Toxicology Program (TR 364). The chemical was given by diet to 50 rats per sex per dose level at 0, 125 and 250 ppm for 103 weeks. The animals were observed twice per day for mortality, morbidity and clinical signs of toxicity. Body weight and food intake were recorded once per week during the first 13 weeks of treatment and monthly thereafter. All animals found dead and those that survived treatment were necropsied.Except for the aorta, vagina, coagulation glands and peripheral nerve, the gross and histopathological examinations were performed as per the latest OECD 408.No treatment-related effects on mortality were observed. Clinical signs were limited to discoloured fur and skin at ≥125 ppm. Male rats treated at 125 and 250 ppm weighed slightly more (about 3-7%) than the control group from week 77 to the end of the study period. Female rats treated at 125 and 250 ppm weighed about the same as the control group throughout the entire study period. No significant effects on food intake were observed. No treatment-related gross pathological effects were observed. Neoplastic findings were limited to increased incidences of keratoacanthomas of the skin (male rats treated at 250 ppm) andpheochromocytomas or malignant pheochromocytomas(female rats treated at 250 ppm). Neither of the two neoplasms were conclusively attributed to the test chemical. No non-neoplastic lesions attributed to the test chemical were observed. The authors of the study acknowledge that the rats could potentially have tolerated a higher dose than 250 ppm. The dose of 250 ppm corresponded to 10 (males) and 12 mg/kg bw/day (females).The study was performed according to GLP.

 

Study 4

The read-across analogueTetrasodium hexacyanoferrate[CAS 13601-19-9; EC: 237-081-9]has been tested for repeated dose toxicity as reported by the European Food Safety Authority (2018). The chemical was given by diet to 48 rats per sex per dose level for a total of two years. Dose levels were equal to about 0, 4.4, 45 and 450.7 mg/kg bw/day (males) and 0, 6.2, 62.5 and 630.1 mg/kg bw/day (females). The animals were observed once per day for mortality, morbidity, and clinical signs of toxicity. Body weight, food intake, and water consumption were recorded initially (body weight only), once per two weeks during the first year, and monthly during the second year. Blood samples were collected from 12 rats per sex per dose level at 14, 26 and 54 weeks of study, and from all surviving rats at 24 months of study. Blood parameters included total red blood cell and white blood cell counts, differential white blood cell counts, haemoglobin concentrations, packed cell volume, and reticulocyte count. Urine samples were collected from ≥10 rats per sex per dose level at 2 months intervals until 24 months of study, at which time urine samples were collected from all surviving rats. Urine samples collected over a 6-hour period were examined for volume, specific gravity, pH, glucose, blood, bilirubin, ketones, and protein. Urine samples collected over a 2-hour period following an oral water load of 25 ml/kg were examined for volume, specific gravity and cell content. Urine samples collected over a 6-hour period following an oral water load of 25 ml/kg were examined for volume and specific gravity.All animals found dead and those that survived treatment were necropsied. The major organs were weighed, and the following organs were examined microscopically for neoplastic and non-neoplastic lesions:adrenal, aorta, bladder, brain, caecum, colon, duodenum, eye, Harderian gland,heart, ileum, kidney, liver, lung, lymph node, mammarygland, muscle, nerve, oesophagus, ovary, pancreas,pituitary, prostate, rectum, salivarygland, seminal vesicles, spinal cord, spleen, stomach, testis, thymus, thyroid,trachea, uterus,and vagina. No treatment-related effects on mortality were observed. Male rats treated at 450.7 mg/kg bw/day showed an increased incidence of pneumonia compared to the control group. This effect was not attributed to the test chemical in the report. No treatment-related effects on clinical signs, body weight, food intake, or haematology were observed. Reduced water intake during the first 9 months of study was observed at 450.7 (males) and 630.1 mg/kg bw/day (females). Except for increased number of cells excreted in the 2-hour urine samples, most notably at mid- and high dose levels, no significant effects in urinalysis were observed. No histopathological effects of toxicological significance were observed. The panel-derived NOAELs were 4.4 (males) and 6.2 mg/kg bw/day (females). The study was performed pre-GLP. 

 

Study 5

The read-across analoguecupric sulfate pentahydrate [CAS: 7758-99-8; EC: 616-477-9]has been tested for repeated dose toxicity by the National Toxicology Program (1993). The chemical was given by diet to 10 rats per sex per dose level at 0, 500, 1000, 2000, 4000 and 8000 ppm. The animals were observed twice per day for mortality and morbidity. Clinical observations were recorded once per week. Body weights were recorded prior to treatment, on day 1 of treatment, and weekly thereafter. Food intake was recorded once per week. Blood samples were collected from all surviving rats at the end of the study period. Haematology parameters included haematocrit, haemoglobin, erythrocytes, reticulocytes, nucleated erythrocytes, mean cell volume, mean cell hemoglobin, mean cell hemoglobin concentration, platelets, white blood cell counts, and differential white blood cell counts. Clinical chemistry parameters includedALT, ALP, 5'-nucleotidase, SDH, bile salts, total protein, albumin, creatinine, and urea nitrogen. Urine samples were collected from all surviving rats at the end of the study period. Urine parameters included creatinine, glucose, protein, AST, NAG, urine volume, and specific gravity. Each animal was given a complete necropsy. Body weights of the following organs were recorded: liver, thymus, right kidney, right testis, heart, lungs, and brain. The following organs were examined grossly and histologically at 0 and ≤8000 ppm: adrenal glands, brain (three sections), oesophagus, eyes (if grossly abnormal), femur with marrow, gross lesions, heart, intestines (large: cecum, colon, rectum; small: duodenum, jejunum, ileum), kidneys, liver, lung/mainstem bronchi, lymph nodes (mandibular, mesenteric), mammary gland, nasal cavity and turbinates (three sections), ovaries, pancreas, parathyroid glands, pharynx (if grossly abnormal), pituitary gland, preputial or clitoral glands, prostate gland, salivary glands, spinal cord/sciatic nerve (if neurologic signs were present), spleen, stomach (forestomach and glandular stomach), testes (with epididymis), thymus, thyroid gland, trachea, urinary bladder, and uterus. In addition to the dosed animals above, supplemental animals (10 rats/sex/dose) treated at 0, 500, 1000, 2000, 4000 and 8000 ppm were included for interim evaluations (on days 5 and 21) of haematology, clinical chemistry, and urinalysis. The supplemental animals treated at 0, 500, 1000, 2000 and 4000 ppm were examined for sperm morphology and vaginal cytology. All animals survived to planned death, apart from one female rat (at 1000 ppm) that was killed by accident. No clinical signs of toxicity attributed to the test chemical were observed. Average terminal body weights relative to the controls were 92, 99, 98, 93 and 76% (males) and 101, 103, 101, 97, and 93% (females) at 500, 1000, 2000, 4000 and 8000 ppm, respectively. No remarkable effects on food intake were observed at 500, 1000, 2000 or 4000 ppm. Food intake by male and female rats treated at 8000 ppm were slightly below that of the controls. Treatment-related effects on haematology, clinical chemistry and urinalysis were, for the most part, restricted to the 4000 and 8000 ppm groups. The observed effects were indicative of hepatocellular damage, of compensatory responses to anaemia by the bone marrow, of and renal tubule epithelial damage. No significant changes in absolute organ weights were observed up to 4000 ppm. Relative organ weights for dosed animals were similar to those of the controls or increased with decreasing mean body weight at 4000 and 8000 ppm. Gross lesions attributed to the test chemical were limited to forestomach lesions at 2000 ppm and above. The histopathological examination revealed an increased incidence of hyperplasia and hyperkeratosis of the forestomach at 2000 ppm and above; an increased incidence of inflammation of the liver at 2000 ppm and above; and an increased incidence in the number (and size) of protein droplets in the epithelial cytoplasm and the lumen of the proximal convoluted tubules at 2000 ppm and above. The mean severity of each histopathological finding at 2000 ppm never exceeded 1.6 according to the grading scale in the study (1=minimal, 2=mild, 3=moderate, and 4=marked). Histopathological findings at 1000 ppm were limited to one incidence of cytoplasmic alteration (kidneys) which was described as minimal in nature. No significant effects on sperm morphology or vaginal cytology were observed. NOAEL for the test chemical was considered at 1000 ppm. The dose of 1000 ppm corresponded to 64 (males) and 68 mg/kg bw/day (females).The study was performed according to GLP.

Repeated Dose Inhalation Toxicity

A short-term toxicity study need not be conducted as because exposure of humans via inhalation route in production and/or use is not likely based on the provided thorough and rigorous exposure assessment. The estimated vapor pressure of the test chemical was 3.49E-10 mm Hg.Hence, this endpoint can be considered for waiver

Repeated Dose dermal Toxicity

A short-term toxicity study need not be conducted as because exposure of humans via dermal route in production and/or use is not likely based on the provided thorough and rigorous exposure assessment.The acute dermal LD50 of the test chemical can be considered to be >2000 mg/kgbw. Hence, this endpoint was considered for waiver.

Justification for classification or non-classification

The chemical is regarded to be classified as Not Classified by the oral, dermal and inhalation route.