Registration Dossier

Administrative data

Description of key information

Repeated dose toxicity: Oral

The No Observed Adverse Effect Level (NOAEL) was considered to be 1000 mg/kg/day, when rats were treated with the given test chemical during repeated dose toxicity study.

 

Repeated dose toxicity: Inhalation

A short-term toxicity study does not need to be conducted because exposure of humans via inhalation in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment. Also, the given test chemical has very low vapor pressure 3.28E-013 Pa (2.46E-015 mm Hg), so the potential for the generation of inhalable vapors is very low. 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 toxicity: Dermal

A short-term toxicity study does not need to be conducted because exposure of humans via dermal in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment. Also, the acute dermal toxicity value for test chemical (as provided in section 7.2.3) is >2000 mg/kg body weight. Given the use of the chemical; repeated exposure by the dermal route is unlikely since the use of gloves is common practice in industries. Thus, it is expected that test chemical shall not exhibit 28 day repeated dose toxicity by the dermal route. In addition, there is no data available that suggests that test chemical shall exhibit repeated dose toxicity by the dermal route. Hence this end point 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
Reference
Endpoint:
sub-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 from various test chemicals
Justification for type of information:
Weight of evidence approach based on the available information from various test chemicals.
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
other: As mentioned below
Principles of method if other than guideline:
WoE report is based on 3 repeated dose toxicity studies i.e. WoE-2, WoE-3 and WoE-4.
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
other: 2. Crj: CD(SD) 3. Wistar 4. Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
2. Age at study initiation of dosing: 10 weeks old
3. No data
4. No data
Route of administration:
oral: gavage
Vehicle:
other: 2. 0.5 % Methylcellulose aqueous solution 3. 1% CMC (carboxymethyl cellulose) aqueous solution 4. water
Details on oral exposure:
2. PREPARATION OF DOSING SOLUTIONS:
The test chemical was dissolved in 0.5 % Methylcellulose aqueous solution to give dose level of 0, 40, 200 or 1000 mg/Kg/day
VEHICLE
- Justification for use and choice of vehicle (if other than water): 0.5 % Methylcellulose aqueous solution
- Concentration in vehicle: 0, 40, 200 or 1000 mg/Kg/day
3. PREPARATION OF DOSING SOLUTIONS:
VEHICLE
- Concentration in vehicle:0, 100, 300 and 1000 mg/kg bw/day (0, 94, 282 or 940 mg active dye/kg bw/day)
- Amount of vehicle (if gavage): 10 ml/kg
4. PREPARATION OF DOSING SOLUTIONS:
VEHICLE
- Justification for use and choice of vehicle (if other than water): Water is used as a vehicle
- Concentration in vehicle: 0, 27, 109 or 435 mg active dye/kg bw/day) in water (5 ml/kg)
- Amount of vehicle (if gavage): 5ml/kg
- Lot/batch no. (if required):2060608
- Purity: 54.4%purity
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The prepared solution was confirmed by UV-visible absorbance method
Duration of treatment / exposure:
2. Male: 42 days / Female: 41 - 45 days (from 14 days before mating to day 4 of lactation)
3. 13 weeks
4. 13 weeks
Frequency of treatment:
Daily
Remarks:
2. 0, 40, 200 or 1000 mg/Kg/day
3. 0, 100, 300 and 1000 mg/kg bw/day (0, 94, 282 or 940 mg active dye/kg bw/day)
4. 0, 50, 200 or 800 mg/kg bw/day (0, 27, 109 or 435 mg active dye/kg bw/day) in water (5 ml/kg) by gavage.
No. of animals per sex per dose:
2. Test group:
0 mg/Kg/day: 7 males and 12 females
40 mg/Kg/day: 12 males and 12 females
200 mg/Kg/day: 12 males and 12 females
1000 mg/Kg/day: 7 males and 12 females

Recovery group:
0 mg/Kg/day: 5 males and 5 females
1000 mg/Kg/day: 5 males and 5 females
3. 10 animals per sex and group
4. 10 animals per sex and group
Control animals:
yes, concurrent vehicle
Details on study design:
2.
- Rationale for selecting satellite groups: 5 females
- Post-exposure recovery period in satellite groups: Females, 14 days
3. - Dose selection rationale: Dose levels were selected on the basis of a previous 14-day oral toxicity study in rats.
4. - Dose selection rationale : Doses were selected on the basis of a previous 14-day oral toxicity study in rats.
Positive control:
No data
Observations and examinations performed and frequency:
2. CAGE SIDE OBSERVATIONS: Yes
- Time schedule: No data
- Cage side observations checked in table [No.?] were included. No data

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): No data
- 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 averages 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: 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. TP, Alb, 2-Glob

URINALYSIS: Yes
- 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: Yes
- 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: No data
- Time schedule for examinations: No data
- How many animals: No data
- Dose groups that were examined: No data
- Parameters checked in table [No.?] were examined. No data

3. CAGE SIDE OBSERVATIONS: Yes
- Time schedule: No data available
- Cage side observations checked in table No data available

DETAILED CLINICAL OBSERVATIONS: Yes

BODY WEIGHT: Yes
- Time schedule:Weekly
- Time schedule for examinations:Daily cage side observations were perform.

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: Food intake is measured at week 13 ,other details are not given.

- 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 averages 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

OPHTHALMOSCOPIC EXAMINATION: Yes

- Time schedule for examinations: During acclimation period

- Dose groups that were examined:All animals in acclimation period and high dose animals in week 13.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Not mentioned
- Anaesthetic used for blood collection: Yes (identity) -No data
- Animals fasted: No data
- How many animals: 10 animals per sex and group for treatment
- Parameters checked in table were examined-No

URINALYSIS: Yes
- Time schedule for collection of urine: Week 13
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table.were examined.- No
NEUROBEHAVIOURAL EXAMINATION: No data

4. DETAILED CLINICAL OBSERVATIONS: Yes / No / No data
- Time schedule:
BODY WEIGHT: No changes in body weight and food intake were reported.
- Time schedule for examinations: Body weights were observed weekly

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 available

- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No food intake related changes reported

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data available

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations: Evaluation were done at the beginning of the treatment period and in week 13.
- Anaesthetic used for blood collection: No data
- Animals fasted: No data available
- How many animals:10 animals per sex and group
- Parameters checked in table -No.


OPHTHALMOSCOPIC EXAMINATION: No ocular findings were observed

- Time schedule for examinations: No data
- Dose groups that were examined:NA

HAEMATOLOGY: Yes
- Time schedule for collection of blood: No details given
- Anaesthetic used for blood collection: No data available
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. No data



CLINICAL CHEMISTRY: No data
- Time schedule for collection of blood: No data available
- Animals fasted: No data available

- How many animals:

- Parameters checked in table were examined-no


URINALYSIS: Yes

Time schedule for collection of urine: No data available

- Metabolism cages used for collection of urine: No data available

- Animals fasted: No data available - Parameters checked in table [No.?] were examined.



CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: No data available
- Animals fasted: No data available
- How many animals: No data available
- Parameters checked in table [No.?] were examined.



NEUROBEHAVIOURAL EXAMINATION: No data available
- Time schedule for examinations: No data available
Sacrifice and pathology:
2. GROSS PATHOLOGY: Yes, no detailed data available
HISTOPATHOLOGY: Yes, no detailed data available
3. GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
4. GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
Other examinations:
No data
Statistics:
No data
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
2. effects observed, treatment-related - Colored feces were noted in male and female rats treated with 40, 200 or 1000 mg/Kg/day
3. no effects observed - There were no adverse clinical signs were observed.
4. no effects observed - No remarkable clinical signs were observed just before death. Increased salivation was observed at all dose levels with a dose-dependent incidence. Loud breathing was noted in 1/10 females given 50 mg/kg bw/day, 1/10 females given 200 mg/kg bw/day, 2/10 females and 1/10 males given 800 mg /kg bw/day. Regurgitation occurred in all treatment groups (10 – 30%).
Mortality:
no mortality observed
Description (incidence):
2. no mortality observed
3. no mortality observed - There were no deaths were observed.
4. no mortality observed - There were no treatment-related deaths.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
2. no effects observed
3. no effects observed - There were no changes in body weights were observed.
4. no effects observed - No treatment related changes observed.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
2. no effects observed
3. no effects observed - There were no changes in food intake were observed.
4. no effects observed - No treatment related changes 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):
2. not specified
3. no effects observed - No substance related ocular findings were reported.
4. no effects observed - No ocular findings were reported.
Haematological findings:
no effects observed
Description (incidence and severity):
2. no effects observed
3. no effects observed - Statistically significant changes in haematological parameters at low dose included increased prothrombin time (low- and high-dose males) and activated partial thromboplastin time (high-dose males).
4. no effects observed - Statistically significant changes in haematological parameters included decreased values for white blood cells (high-dose males), leukocytes, (high-dose males), prothrombin time (highdose
females) and fibrinogen (high-dose females); and increased values for packed cell volume (high-dose females), mean cell volume (mid- and high-dose females), mean cell haemoglobin (mid-dose females), mean cell haemoglobin concentration (high-dose males), prothrombin time (high-dose males) and activated partial thromboplastin time (high-dose males).
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
2. effects observed, treatment-related - Decrease in the TP, decrease in the Alb and increase in the percentage of alpha 2-Glob (Male) in 1000 mg/Kgday treated animals. Though albumin level (mg/dL) was significantly decreased in males of the high dose group, A/G ratio was not affected.
3. no effects observed - The changes observed in blood clinical chemistry laboratory parameters were considered to be of no toxicological significance.
4. no effects observed - Statistically significant changes in blood clinical chemistry parameters included slight decreases in inorganic phosphorus (males in all dose groups), glucose (mid-dose males and high-dosefemales), urea (high-dose males), alkaline phosphatase (high-dose males) and alanine aminotransferase (high-dose males). These changes were considered to be of no toxicological importance.
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
2. effects observed, treatment-related - Abnormal color of urine was observed in males and females treated with 200 or 1000 mg/Kg/day
3. no effects observed - The changes observed in urinalysis were considered to be of no toxicological significance.
4. no effects observed - The only finding in the urinalysis was an increased pH of urine in females (highdose).
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
2. no effects observed
3. effects observed, non-treatment-related - In males given 1000 mg/kg bw/day, slightly increased locomotor activity was observed after 15 minutes; in absence of a clear dose-relationship this finding was considered to be fortuitous. Statistically significant decreased locomotor activity was observed after 45 minuts in females given 300 or 1000 mg/kg bw/day and persisted until the end of the measurement period in females given 1000 mg/kg bw/day. The toxicological significance was unclear, but in absence of this finding in males and in absence of similar parameters (general) a test article relation was considered to be unlikely. Changes in grip strength were reported in treated males and females. However, data did not show dose-relationship and no correlation between fore- and hind-limb strength. Effects therefore might not be related to the test substance.
4. not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
2. no effects observed
3. no effects observed - Statistically significant changes in relative organ weights included increased kidney weight in low- and mid-dose males, and decreased heart weight in mid-dose females. In absence of any dose relationship these findings were considered to be fortuitous.
4. no effects observed - There were no significant differences in organ weights between treated and control groups.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
2. effects observed, treatment-related - Colored aqueous content in the alimentary tract in 40 mg/Kgday treated male animals. Colored aqueous content in the alimentary tract (Male/Female), Mucosal discoloration of alimentary tract (Male) in 200 mg/Kg/day treated animals. Colored aqueous content in the alimentary tract (Male/Female), Mucosal discoloration of alimentary tract (Male/Female) in 1000 mg/Kg/day treated animals.
3. effects observed, non-treatment-related - Dark blue faeces were observed at all dose levels and were related to the staining properties of the test substance. The only findings at necropsy were a blue discolouration of the mucosal surface of the stomach and/or intestines observed for a few animals given 1000 mg/kg bw/day.
4. effects observed, treatment-related - Findings at necropsy were dose-related greenish contents or greenish colorations of the mucosa of the digestive tract. At the microscopic examination, alveolar oedema together with pigmented granular material was noted. Coloured urine, faeces, fur and extremities were observed at all dose levels and were related to the staining properties of the test chemical.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
2. no effects observed
3. effects observed, non-treatment-related - The histopathological changes noted at the microscopic examination of tissues and organs of treated animals were similar in incidence, severity and morphological characteristics to those observed in the control group and therefore, not considered to be treatment-related.
4. no effects observed - The histopathological changes noted at the microscopic examination of tissues and organs of treated animals were similar in incidence, severity and morphological characteristics to those observed in the control group and therefore, not considered to be treatment-related.
Histopathological findings: neoplastic:
not specified
Other effects:
effects observed, treatment-related
Description (incidence and severity):
4. The only finding considered to be treatment-related was the increased activated partial thromboplastin time in high-dose males (+30% in mean).
Dose descriptor:
NOAEL
Remarks:
2
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
haematology
clinical biochemistry
gross pathology
histopathology: non-neoplastic
Remarks on result:
other: No significant effects were observed at 1000 mg/Kg/day
Dose descriptor:
NOAEL
Remarks:
3
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
ophthalmological examination
haematology
clinical biochemistry
urinalysis
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
Remarks on result:
other: No effect observed
Dose descriptor:
NOAEL
Remarks:
4
Effect level:
200 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
haematology
clinical biochemistry
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
Remarks on result:
other: No effect observed
Critical effects observed:
not specified
Conclusions:
The No Observed Adverse Effect Level (NOAEL) was considered to be 1000 mg/kg/day, when rats were treated with the given test chemical during repeated dose toxicity study.
Executive summary:

Data available from the various sources was reviewed to determine the toxic nature of the given test chemical. The studies are as mentioned below:

 

Combined Repeated Dose Toxicity Study with the Reproduction/ Developmental Toxicity Screening Test (OECD TG422) was performed to determine the toxic nature of the test chemical. The study was performed using Crl:CD (SD) male and female rats. Recovery group was also included in the study. The test chemical was dissolved in 0.5 % methylcellulose aqueous solution and used at dose level of 0, 40, 200 or 1000 mg/Kg/day. The treated animals were observed for mortality, clinical signs, changes in body weight, food consumption, urinalysis, hematology, clinical chemistry and were subjected to gross and histopathology. No mortality was noted and no effects were observed in clinical signs, functional battery observations, body weight and food consumption changes, hematology, organ weights and histopathology. Colored feces (40, 200 and 1000 mg/Kg/day) and abnormal urine color (200 and 1000 mg/Kg/day) was noted in the treated animals. Decrease in the TP, decrease in the Alb and increase in the percentage of alpha 2-Glob (Male) in treated animals of 1000 mg/Kg/day. Though albumin level (mg/dL) was significantly decreased in males of the high dose group, A/G ratio was not affected. Colored aqueous content in the alimentary tract in 40 mg/Kgday treated male animals. Colored aqueous content in the alimentary tract (Male/Female), mucosal discoloration of alimentary tract (Male) in 200 mg/Kg/day treated animals. Colored aqueous content in the alimentary tract (Male/Female), mucosal discoloration of alimentary tract (Male/Female) in 1000 mg/Kg/day treated animals. Based on the observations of the study, the no observed adverse effect level (NOAEL) for the test chemical is considered to be 1000 mg/Kg/day when male and female rats were exposed to the test chemical in combined Repeated Dose Toxicity Study with the Reproduction/ Developmental Toxicity Screening Test (OECD TG422). 

 

In another repeated dose oral study, the given test chemical was assessed for its possible toxic potential. For this purpose sub-chronic toxicity study was conducted in 10 animals per sex and group Wistar rats. Daily oral gavage at 0 (vehicle, 1% aqueous solution of carboxymethylcellulose), 100, 300 and 1000 mg/kg bw/day (0, 94, 282 or 940 mg active dye/kg/day) at a dosing volume of 10 ml/kg was performed for 13 weeks. These dose levels were selected on the basis of a previous 14-day oral toxicity study in rats. Evaluations and measurements included mortality, daily cage-side observations, weekly body weight, food intake and detailed clinical observations, ophthalmoscopy (on all animals in acclimation period and on control and high dose animals in week 13), as well as functional parameters, hematology, blood clinical chemistry and urinalysis (week 13). At the end of the dosing period, animals were killed and subjected to macroscopic examination, selected organs were weighed, and organs/tissues were preserved. Microscopic examination was performed for specified tissues/organs from control and high dose rats, and for gross anomalies from all animals. There were no deaths, no adverse clinical signs or changes in body weights and food intake. No substance related ocular findings were reported. Dark blue faeces were observed at all dose levels and were related to the staining properties of the test substance. In males given 1000 mg/kg bw/day, slightly increased locomotor activity was observed after 15 minutes; in absence of a clear dose-relationship this finding was considered to be fortuitous. Statistically significant decreased locomotor activity was observed after 45 minuts in females given 300 or 1000 mg/kg bw/day and persisted until the end of the measurement period in females given 1000 mg/kg bw/day. The toxicological significance was unclear, but in absence of this finding in males and in absence of similar parameters (general and detailed clinical observations) a test article relation was considered to be unlikely. Changes in grip strength were reported in treated males and females. However, data did not show dose-relationship and no correlation between fore- and hind-limb strength. Effects therefore might not be related to the test substance. Statistically significant changes in haematological parameters at low dose included increased prothrombin time (low- and high-dose males) and activated partial thromboplastin time (high-dose males). The changes observed in blood clinical chemistry laboratory parameters and in urinalysis were considered by the study authors to be of no toxicological significance. Statistically significant changes in relative organ weights included increased kidney weight in low- and mid-dose males, and decreased heart weight in mid-dose females. In absence of any dose relationship these findings were considered to be fortuitous. The only findings at necropsy were a blue discolouration of the mucosal surface of the stomach and/or intestines observed for a few animals given 1000 mg/kg bw/day. The histopathological changes noted at the microscopic examination of tissues and organs of treated animals were similar in incidence, severity and morphological characteristics to those observed in the control group and therefore, not considered to be treatment-related. Under the condition of the study, the No Observed Adverse Effect Level (NOAEL) was considered to be 1000 mg/kg bw/day (940 mg active dye/kg bw/day).

 

Both the above study results were supported with the sub-chronic toxicity study conducted by using the given test chemical in 10 animals per sex and group Sprague-Dawley rats at the doses 0, 50, 200 or 800 mg/kg bw/day (0, 27, 109 or 435 mg active dye/kg bw/day) in water (5 ml/kg) by gavage route 13 weeks. These doses were selected on the basis of a previous 14-day oral toxicity study in rats. Evaluations and measurements included mortality, daily clinical observations, weekly body weight and food intake, ophthalmoscopy (in acclimation once before the beginning of the treatment period and in week 13 on control and high dose animals), hematology, blood clinical chemistry and urinalysis (week 13). At the end of the dosing period, surviving animals were killed and subjected to a complete macroscopic examination, principal organs (adrenals, heart, kidneys, liver, ovaries, spleen, testes, thymus) were weighed, and a full spectrum of tissues were preserved. Microscopic examination was performed for specified tissues/organs from all decedent rats, control and high dose rats killed at the end of the study, as well as for gross anomalies, lungs, liver and kidneys from all animals. There were no treatment-related deaths. Two treated females (one given 200 mg/kg bw/day and one given 800 mg/kg bw/day) died during the study. Purplish colour and/or purplish and blue contents were observed in the lungs, in buccal and thoracal cavities and in the trachea of these females. No remarkable clinical signs were observed just before death; at the microscopic examination, alveolar oedema together with pigmented granular material was noted. Their death, therefore, to a gavage error and not to the administration of the test substance. Increased salivation was observed at all dose levels with a dose-dependent incidence. Loud breathing was noted in 1/10 females given 50 mg/kg bw/day, 1/10 females given 200 mg/kg bw/day, 2/10 females and 1/10 males given 800 mg /kg bw/day. Regurgitation occurred in all treatment groups (10 –30%). Coloured urine, faeces, fur and extremities were observed at all dose levels and were related to the staining properties of Jarocol Violet 43. No other clinical signs, no ocular findings or changes in body weight and food intake were reported. There were no significant differences in organ weights between treated and control groups. Findings at necropsy were dose-related greenish contents or greenish colorations of the mucosa of the digestive tract. The histopathological changes noted at the microscopic examination of tissues and organs of treated animals were similar in incidence, severity and morphological characteristics to those observed in the control group and therefore, not considered by the study authors to be treatment-related. Statistically significant changes in blood clinical chemistry parameters included slight decreases in inorganic phosphorus (males in all dose groups), glucose (mid-dose males and high-dose females), urea (high-dose males), alkaline phosphatase (high-dose males) and alanine aminotransferase (high-dose males). These changes were considered to be of no toxicological importance. Statistically significant changes in haematological parameters included decreased values for white blood cells (high-dose males), leukocytes, (high-dose males), prothrombin time (highdose females) and fibrinogen (high-dose females); and increased values for packed cell volume (high-dose females), mean cell volume (mid- and high-dose females), mean cell haemoglobin (mid-dose females), mean cell haemoglobin concentration (high-dose males), prothrombin time (high-dose males) and activated partial thromboplastin time (high-dose males). The only finding in the urinalysis was an increased pH of urine in females (highdose). The only finding considered to be treatment-related was the increased activated partial thromboplastin time in high-dose males (+30% in mean). Under the condition of the study, the No Observed Adverse Effect Level was considered to be 200 mg/kg bw/day (109 mg active dye/kg bw/day) based on the increased activated partial thromboplastin time in high-dose males.

 

Thus based on the above studies, the No Observed Adverse Effect Level (NOAEL) was considered to be 1000 mg/kg body weight and hence is not likely to classify as per the criteria mentioned in CLP regulation.

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:
Data is Klimicsh 2 and from handbook or collection of data.

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:
a short-term toxicity study does not need to be conducted because exposure of humans via inhalation in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment
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:
a short-term toxicity study does not need to be conducted because exposure of humans via the dermal route in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment
Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
Waiver

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Repeated dose toxicity: Oral

Data available from the various sources was reviewed to determine the toxic nature of the given test chemical. The studies are as mentioned below:

 

Combined Repeated Dose Toxicity Study with the Reproduction/ Developmental Toxicity Screening Test (OECD TG422) was performed to determine the toxic nature of the test chemical. The study was performed using Crl:CD (SD) male and female rats. Recovery group was also included in the study. The test chemical was dissolved in 0.5 % methylcellulose aqueous solution and used at dose level of 0, 40, 200 or 1000 mg/Kg/day. The treated animals were observed for mortality, clinical signs, changes in body weight, food consumption, urinalysis, hematology, clinical chemistry and were subjected to gross and histopathology. No mortality was noted and no effects were observed in clinical signs, functional battery observations, body weight and food consumption changes, hematology, organ weights and histopathology. Colored feces (40, 200 and 1000 mg/Kg/day) and abnormal urine color (200 and 1000 mg/Kg/day) was noted in the treated animals. Decrease in the TP, decrease in the Alb and increase in the percentage of alpha 2-Glob (Male) in treated animals of 1000 mg/Kg/day. Though albumin level (mg/dL) was significantly decreased in males of the high dose group, A/G ratio was not affected. Colored aqueous content in the alimentary tract in 40 mg/Kgday treated male animals. Colored aqueous content in the alimentary tract (Male/Female), mucosal discoloration of alimentary tract (Male) in 200 mg/Kg/day treated animals. Colored aqueous content in the alimentary tract (Male/Female), mucosal discoloration of alimentary tract (Male/Female) in 1000 mg/Kg/day treated animals. Based on the observations of the study, the no observed adverse effect level (NOAEL) for the test chemical is considered to be 1000 mg/Kg/day when male and female rats were exposed to the test chemical in combined Repeated Dose Toxicity Study with the Reproduction/ Developmental Toxicity Screening Test (OECD TG422). 

 

In another repeated dose oral study, the given test chemical was assessed for its possible toxic potential. For this purpose sub-chronic toxicity study was conducted in 10 animals per sex and group Wistar rats. Daily oral gavage at 0 (vehicle, 1% aqueous solution of carboxymethylcellulose), 100, 300 and 1000 mg/kg bw/day (0, 94, 282 or 940 mg active dye/kg/day) at a dosing volume of 10 ml/kg was performed for 13 weeks. These dose levels were selected on the basis of a previous 14-day oral toxicity study in rats. Evaluations and measurements included mortality, daily cage-side observations, weekly body weight, food intake and detailed clinical observations, ophthalmoscopy (on all animals in acclimation period and on control and high dose animals in week 13), as well as functional parameters, hematology, blood clinical chemistry and urinalysis (week 13). At the end of the dosing period, animals were killed and subjected to macroscopic examination, selected organs were weighed, and organs/tissues were preserved. Microscopic examination was performed for specified tissues/organs from control and high dose rats, and for gross anomalies from all animals. There were no deaths, no adverse clinical signs or changes in body weights and food intake. No substance related ocular findings were reported. Dark blue faeces were observed at all dose levels and were related to the staining properties of the test substance. In males given 1000 mg/kg bw/day, slightly increased locomotor activity was observed after 15 minutes; in absence of a clear dose-relationship this finding was considered to be fortuitous. Statistically significant decreased locomotor activity was observed after 45 minuts in females given 300 or 1000 mg/kg bw/day and persisted until the end of the measurement period in females given 1000 mg/kg bw/day. The toxicological significance was unclear, but in absence of this finding in males and in absence of similar parameters (general and detailed clinical observations) a test article relation was considered to be unlikely. Changes in grip strength were reported in treated males and females. However, data did not show dose-relationship and no correlation between fore- and hind-limb strength. Effects therefore might not be related to the test substance. Statistically significant changes in haematological parameters at low dose included increased prothrombin time (low- and high-dose males) and activated partial thromboplastin time (high-dose males). The changes observed in blood clinical chemistry laboratory parameters and in urinalysis were considered by the study authors to be of no toxicological significance. Statistically significant changes in relative organ weights included increased kidney weight in low- and mid-dose males, and decreased heart weight in mid-dose females. In absence of any dose relationship these findings were considered to be fortuitous. The only findings at necropsy were a blue discolouration of the mucosal surface of the stomach and/or intestines observed for a few animals given 1000 mg/kg bw/day. The histopathological changes noted at the microscopic examination of tissues and organs of treated animals were similar in incidence, severity and morphological characteristics to those observed in the control group and therefore, not considered to be treatment-related. Under the condition of the study, the No Observed Adverse Effect Level (NOAEL) was considered to be 1000 mg/kg bw/day (940 mg active dye/kg bw/day).

 

Both the above study results were supported with the sub-chronic toxicity study conducted by using the given test chemical in 10 animals per sex and group Sprague-Dawley rats at the doses 0, 50, 200 or 800 mg/kg bw/day (0, 27, 109 or 435 mg active dye/kg bw/day) in water (5 ml/kg) by gavage route 13 weeks. These doses were selected on the basis of a previous 14-day oral toxicity study in rats. Evaluations and measurements included mortality, daily clinical observations, weekly body weight and food intake, ophthalmoscopy (in acclimation once before the beginning of the treatment period and in week 13 on control and high dose animals), hematology, blood clinical chemistry and urinalysis (week 13). At the end of the dosing period, surviving animals were killed and subjected to a complete macroscopic examination, principal organs (adrenals, heart, kidneys, liver, ovaries, spleen, testes, thymus) were weighed, and a full spectrum of tissues were preserved. Microscopic examination was performed for specified tissues/organs from all decedent rats, control and high dose rats killed at the end of the study, as well as for gross anomalies, lungs, liver and kidneys from all animals. There were no treatment-related deaths. Two treated females (one given 200 mg/kg bw/day and one given 800 mg/kg bw/day) died during the study. Purplish colour and/or purplish and blue contents were observed in the lungs, in buccal and thoracal cavities and in the trachea of these females. No remarkable clinical signs were observed just before death; at the microscopic examination, alveolar oedema together with pigmented granular material was noted. Their death, therefore, to a gavage error and not to the administration of the test substance. Increased salivation was observed at all dose levels with a dose-dependent incidence. Loud breathing was noted in 1/10 females given 50 mg/kg bw/day, 1/10 females given 200 mg/kg bw/day, 2/10 females and 1/10 males given 800 mg /kg bw/day. Regurgitation occurred in all treatment groups (10 –30%). Coloured urine, faeces, fur and extremities were observed at all dose levels and were related to the staining properties of Jarocol Violet 43. No other clinical signs, no ocular findings or changes in body weight and food intake were reported. There were no significant differences in organ weights between treated and control groups. Findings at necropsy were dose-related greenish contents or greenish colorations of the mucosa of the digestive tract. The histopathological changes noted at the microscopic examination of tissues and organs of treated animals were similar in incidence, severity and morphological characteristics to those observed in the control group and therefore, not considered by the study authors to be treatment-related. Statistically significant changes in blood clinical chemistry parameters included slight decreases in inorganic phosphorus (males in all dose groups), glucose (mid-dose males and high-dose females), urea (high-dose males), alkaline phosphatase (high-dose males) and alanine aminotransferase (high-dose males). These changes were considered to be of no toxicological importance. Statistically significant changes in haematological parameters included decreased values for white blood cells (high-dose males), leukocytes, (high-dose males), prothrombin time (highdose females) and fibrinogen (high-dose females); and increased values for packed cell volume (high-dose females), mean cell volume (mid- and high-dose females), mean cell haemoglobin (mid-dose females), mean cell haemoglobin concentration (high-dose males), prothrombin time (high-dose males) and activated partial thromboplastin time (high-dose males). The only finding in the urinalysis was an increased pH of urine in females (highdose). The only finding considered to be treatment-related was the increased activated partial thromboplastin time in high-dose males (+30% in mean). Under the condition of the study, the No Observed Adverse Effect Level was considered to be 200 mg/kg bw/day (109 mg active dye/kg bw/day) based on the increased activated partial thromboplastin time in high-dose males.

 

Thus based on the above studies, the No Observed Adverse Effect Level (NOAEL) was considered to be 1000 mg/kg body weight and hence is not likely to classify as per the criteria mentioned in CLP regulation.

 

Repeated dose toxicity: Inhalation

A short-term toxicity study does not need to be conducted because exposure of humans via inhalation in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment. Also, the given test chemical has very low vapor pressure 3.28E-013 Pa (2.46E-015 mm Hg), so the potential for the generation of inhalable vapors is very low. 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 toxicity: Dermal

A short-term toxicity study does not need to be conducted because exposure of humans via dermal in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment. Also, the acute dermal toxicity value for test chemical (as provided in section 7.2.3) is >2000 mg/kg body weight. Given the use of the chemical; repeated exposure by the dermal route is unlikely since the use of gloves is common practice in industries. Thus, it is expected that test chemical shall not exhibit 28 day repeated dose toxicity by the dermal route. In addition, there is no data available that suggests that test chemical shall exhibit repeated dose toxicity by the dermal route. Hence this end point was considered for waiver.

 

Justification for classification or non-classification

Based on the data available and applying the weight of evidence approach, the given test chemical does not exhibit toxic nature upon repeated exposure by oral route of exposure. Hence, it is not likely to classify as toxic as per the criteria mentioned in CLP regulation. For repeated inhalation and repeated dermal toxicity wavier was added so, not possible to classify.