Registration Dossier

Administrative data

Description of key information

Repeated dose toxcity: Oral

Based on the available results and applying the weight of evidence approach, the NOAEL for the test chemical can be considered to lie between 40-180 mg/kg/day.

Repeated dose toxicity: Inhalation

A short term toxicity study does not need to be conducted because exposure of humans via inhalation route is highly unlikely based on the thorough and rigorous exposure assessment. The test chemical has very low vapor pressure of 2.61E-10 Pa which is equivalent to 1.96E-12 mm Hg at 25 ° C, so the potential for the generation of inhalable vapour is low. Also the normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be unlikely to occur, and therefore this end point was considered for waiver

Repeated dose toxicity: Dermal

A short term toxicity study does not need to be conducted because exposure of humans via inhalation route is highly unlikely based on the thorough and rigorous exposure assessment. In accordance with ANNEX VII Colum 2 of the REACH regulation, the study need not be conducted if the substance is a strong acid (pH<=2.0) or strong base (pH=> 11.5). The experimental pH of the test chemical is 14. Hence, based on the high pH of the test chemical, the repeated dermal toxicity study 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:
Weight of evidence approach based on various test chemicals
Justification for type of information:
Weight of evidence approach based on various test chemicals
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
other: Weight of evidence approach based on various test chemicals
Principles of method if other than guideline:
Weight of evidence approach based on various test chemicals
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Details on species / strain selection:
not specified
Sex:
male/female
Details on test animals and environmental conditions:
2. TEST ANIMALS
- Source: IGS, Japan Chiyarusu-Lipa Inc., Atsugi breeding center.
- Age at study initiation: 10 weeks old
- Weight at study initiation: Male 389 to 452 g
Female 219 to 266 g
- Fasting period before study: No data available
- Housing: Animals were housed one per by sex in a metal net cage, during the mating period two per sex per cage while mother were housed individually in plastic cage.
- Diet (e.g. ad libitum): Solid feed (NMF, Oriental Yeast Co., Ltd.), ad libitum.
- Water (e.g. ad libitum): Drinking water (NMF, Oriental Yeast Co., Ltd.), ad libitum.
- Acclimatization period: 19 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 3 ℃
- Humidity (%):50 ± 20 %
- Air changes (per hr): 10-15 times per hour ventilation.
- Photoperiod (hrs dark / hrs light): 12 hours dark / 12 hours light
3. - Age at study initiation: 25-38 days old
- Weight at study initiation:
Males: 102-143 g
Females: 95-122 g
Route of administration:
other: 2. oral: gavage; 3. oral: feed
Details on route of administration:
No data
Vehicle:
other: 2. water; 3. diet
Details on oral exposure:
2.
Details on oral exposure
PREPARATION OF DOSING SOLUTIONS: No data available
DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): Purified water
- Concentration in vehicle: 0, 60, 180 or 600 mg/kg body weight/day
- Amount of vehicle (if gavage): 10 ml/ kg body weight dose volume
- Lot/batch no. (if required): OU06G, Tokyo Chemical Industry Co
3. PREPARATION OF DOSING SOLUTIONS: The test chemical was mixed with feed at dose level of 0, 40, 200 and 1000 mg/kg/day

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

VEHICLE
- Justification for use and choice of vehicle (if other than water): Feed
- Concentration in vehicle: 0, 40, 200 and 1000 mg/kg/day
- Amount of vehicle (if gavage): No data
- Lot/batch no. (if required): No data
- Purity: No data
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
3. Actual dose received:
39.9, 198.3 and 1007 mg/kg/day for males
39.1, 206.5 and 1045 mg/kg/day for females
Duration of treatment / exposure:
2. 42 days
3. 28 days
Frequency of treatment:
2,3. daily
Remarks:
Study 2:
Doses / Concentrations:
0, 60, 180 or 600 mg/kg body weight/day
Basis:
actual ingested
Remarks:
Study 3: 0, 40, 200 and 1000 mg/kg/day
No. of animals per sex per dose:
Study 2: Total animals: 101
Control: 12 males, 12 females
60 mg/kg/day: 12 males, 12 females
180 mg/kg/day: 12 males, 12 females
600 mg/kg/day: 12 males, 12 females
With 5 male (out of 12) from each group for recovery and 5 female as satellite group.
3. Total: 24 males and 24 females
0 mg/Kg/day: 6 males and 6 females
40 mg/Kg/day: 6 males and 6 females
200 mg/Kg/day: 6 males and 6 females
1000 mg/Kg/day: 6 males and 6 females
Control animals:
yes, concurrent vehicle
Details on study design:
2. - Dose selection rationale:
14-day repeated oral dose toxicity study using rats of the test chemical (dose setting test)" was carried out earlier dose (dose: 100,300,1000 mg / kg). 1 rat died on the 14th administration in male 1000 mg / kg dose group, in a general state of deaths loose stools (reddish brown), body weight and food consumption was incresed in 300 mg/kg, liver weight was observed high, low value of the ALP, high total protein, expansion of the cecum at autopsy in male and female were observed at 1000 mg/kg daoe gruop. Therefore, the dose level of 0, 60, 180 and 600 mg/kg/bw/day was chosen in the current study.
- Rationale for animal assignment (if not random): Animals were randomized by body weight.
- Rationale for selecting satellite groups: 5 female as satellite group.
- Post-exposure recovery period in satellite groups: No data available
- Section schedule rationale (if not random): No data available
3. Details on study design
- Dose selection rationale: Groups of rats (three males and three females) were administered the test substance continuously in the diet for 14 days at concentrations of 500, 2500, 5000 and 10000 ppm. A group of control rats (three males and three females) received untreated diet of the same batch at the same frequency as treated rats. Animals were group housed by sex and d1ose group, three per cage. Rats were approximately four to five weeks old at study initiation. Males and females weighed 99 to 135 and 100 to 121 grams, respectively, at study initiation. Rats were observed twice daily throughout the treatment period for evidence of reaction to treatment or ill health. A detailed weekly examination was conducted, including palpation. Food consumption was calculated weekly for each group. Body weights were recorded on the day that treatment commenced, at twice weekly intervals during the treatment period and immediately before necropsy. At the end of the 14-day treatment period, all rats were sacrificed and subjected to a detailed necropsy. The following organs were weighed: adrenals, kidneys and liver. The actual doses received were 70.86, 386.9, 764.4 and 1470 mg/kg/day for the males and 71.45, 348.2, 674.2 and 1347 mg/kg/day. All rats survived until scheduled sacrifice and no signs of reaction to treatment were observed during the treatment period. Food consumption, body weight gains and food conversion ratios were unaffected by treatment. Organ weight analysis did not reveal any changes related to treatment with the test substance. Macroscopic pathology findings were unremarkable. It was concluded that administration of the test substance at dietary concentrations of 500, 2500, 5000 or 10000 ppm did not produce any evidence of toxicity. Based on the results from this study, doses of 40, 200 and 1000 mg/kg/day were selected for the four-week dietary study.
- Rationale for animal assignment (if not random): Animals were group housed by sex and dose group
- Rationale for selecting satellite groups: No data
- Post-exposure recovery period in satellite groups: No data
- Section schedule rationale (if not random): No data
Positive control:
not specified
Observations and examinations performed and frequency:
2. CAGE SIDE OBSERVATIONS: Yes
Attitude, fistula attack, abnormal behaviour were observed.

- Time schedule: Once, once a week (but 1,7,14 and 20 days of gestation, mating confirmation female animals nursing the 4th calving animals) were observed.

- Cage side observations checked in table [No.?] were included: On 1,7,14 and 20 days of gestation, mating confirmation female animals nursing the 4th calving animals were observed.

DETAILED CLINICAL OBSERVATIONS: Yes
Pupil diameter, abnormal breathing, the state of the fur-skin, secretions of the eyes, nose, exophthalmos,flair closed state, the visible mucous membranes, autonomic function and urination were observed.

- Time schedule: 3 times daily

BODY WEIGHT: Yes
Throughout the recovery period and the period of administration.

- Time schedule for examinations:
At a frequency of twice a week, check mating female animals were weighed on the 4th and lactation, as well as 0 and 20 days 0, 4, 7, 11, 14, 17 pregnancy.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data available

- 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 data available

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 available

- Time schedule for examinations: No data available

OPHTHALMOSCOPIC EXAMINATION: Yes
Pupil diameter and secretions of the eyes were observed.

- Time schedule for examinations: Daily

- Dose groups that were examined: 0, 60, 180 and 600 mg/kg body weight/day

HAEMATOLOGY: Yes

- Time schedule for collection of blood:
Before autopsy.

- Anaesthetic used for blood collection: Yes

- Animals fasted: Yes
On day 43, overnight (16-20 hours) fasted.
- How many animals: each 5 per sex per group

- Parameters checked in table [No.?] were examined. : RBC, Hb, Ht, MCV, MCH, MCHC, Reticulocyte, Platelet, PT, APTT, Fibrinogen, WBC and differential leucocytes were examined.

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

URINALYSIS: Yes
Urinalysis of male rat were observed on administration of Tetrabutylammonium bromide and the last week of recovery, urine collected from 5 animals in each group and observed the color, pH, protein, ketone bodies, glucose, occult blood, bilirubin, urobilinogen and urine volume.

- Time schedule for collection of urine: Daily 20 hour.

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

- Animals fasted: Yes
For 4 hours

- Parameters checked in table [No.?] were examined.:
Color, pH, protein, ketone bodies, glucose, occult blood, bilirubin, urobilinogen and urine volume were checked.

NEUROBEHAVIOURAL EXAMINATION: No data available

- Time schedule for examinations: No data available

- Dose groups that were examined: n No data available

- Battery of functions tested: sensory activity / grip strength / motor activity / other:
Grip strength: Grip strength measurement of hindlimb and forelimb were tested.

Spontaneous momentum: Spontaneous movement sensor was counting the momentum of 60 minutes and 10 minute intervals.

OTHER: Auditory response, approaching response, contact reaction, pain reaction, anti-morphism air righting reflex, landing leg width was observed.
3. Observations and examinations performed & frequency
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily for health and weekly for palpation
- Cage side observations checked in table [No.?] were included. Rats were observed twice daily throughout the treatment period for evidence of reaction to treatment or ill health. A detailed weekly examination was conducted, including palpation.

DETAILED CLINICAL OBSERVATIONS: No data
- Time schedule: No data

BODY WEIGHT: Yes
- Time schedule for examinations: Body weights were recorded on the day that treatment commenced, at twice weekly intervals during the treatment period and immediately before necropsy.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes, weekly
- 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): Yes / No / No data
- Time schedule for examinations:

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: On Day 28 of treatment, prior to sacrifice
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes, overnight fasting
- How many animals: All animals
- Parameters checked in table [No.?] were examined. Packed cell volume, hemoglobin concentration, erythrocyte count, mean cell hemoglobin concentration, mean cell volume, mean cell hemoglobin and total and differential leukocyte count.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: On Day 28 of treatment, prior to sacrifice
- Animals fasted: Yes, overnight
- How many animals: All animals
- Parameters checked in table [No.?] were examined. alkaline phosphatase activity, alanine amino-transferase activity, aspartate amino-transferase activity, urea concentration, glucose concentration, total bilirubin concentration, creatinine concentration, total protein concentration, electrophoretic protein fractions and sodium, potassium, chloride, calcium and inorganic phosphorus concentrations

URINALYSIS: No data
- Time schedule for collection of urine: No data
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.?] were examined. No data

NEUROBEHAVIOURAL EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
- Battery of functions tested: No data sensory activity / grip strength / motor activity / other: No data

OTHER: No data
Sacrifice and pathology:
2. Sacrifice and pathology
GROSS PATHOLOGY: Yes
Change attributed in intestinal tracts of male and female of 180 and 600 mg/kg/bw/day group and in liver of male and female of 600 mg/kg/bw/day group.

HISTOPATHOLOGY: Yes
Diffuse hyperplasis in mucosa in cecum,which thought to be related to the macroscopic dilation in lunina of cecum, in the rectum, the debris in crypt were observed in female of 600 mg/kg/bw/day group.
In recovery group of 600 mg/kg/bw/day cell infertilation in mucosa were observed cecum,colon and rectum. In the liver , hypertropy of preiobular hepatocyte were observed.
3. Sacrifice and pathology
GROSS PATHOLOGY: Yes, at the end of the 28-day treatment period, all rats were sacrificed and subjected to a detailed necropsy. The following organs were weighed: adrenals, kidneys, heart, spleen, testes and liver.

HISTOPATHOLOGY: Yes, adrenals, kidneys, heart, spleen, lymph node (mesenteric) and liver were examined microscopically.
Statistics:
2.
Statistical analysis was carried out by using x 2 test with continuity correction of Yeates, expected frequency was observed cell of 5 or less, by the direct probability calculation method of Fisher test (significance level for the 4th). Dunnett type test (mean rank test method) (significance level 0,06 ) performed for homogeneity of variance of each group and Bartlett method firstly test was applied. (significance level 0.01, both sides)
3. Student's t-test, Dunnett's test or Fisher's exact probability test
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
2. Before death, the male showed fractures of incisors, soft stool and a decreased in the amount of feces produced. The female showed staining of lower abdominal fur and staggering gait before death.
3. No signs of reaction to treatment were observed during the treatment period
Mortality:
mortality observed, treatment-related
Description (incidence):
2. Deaths occurred in 1 male and 1 pregnant female when treated with 600 mg/kg/day.
3. No mortality was noted in the treated animals
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
2. When treated with 600 mg/kg/day, the males showed suppressed body weight gain while females showed a tendency toward high values in body weight during the administration period. In comparison to control, males in the 600 mg/kg/day group showed a low value in body weight during the recovery period.
3. Body weight gain was unaffected by treatment
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
2. Females showed a tendency toward high values in food consumption during the administration period. A high value in food consumption was also observed in females treated with 180 mg/kg/day.
3. Food consumption and food utilization (amount of food consumed per unit of bodyweight gain) were unaffected by treatment
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Description (incidence and severity):
2. Males treated with 600 mg/kg/day showed a slight decrease in platelets. Similar effects were observed during the recovery period. An increased level of fibrinogen was also seen in the same animals, i.e. the 600 mg/kg/day-treated males and males from the recovery period. Males in recovery period also showed decreased levels of lymphocyte and eosinophils while the level of neutrophils and monocytes were increased. Females treated with 600 mg/kg/day showed a slight increase in RBC and a slight decrease in platelets. These effects were not observed during the recovery period.In addition, a decreased level of fibrinogen was also seen in the same animals, i.e. the 600 mg/kg/day-treated females.Females in recovery period showed decreased levels of lymphocytes and increased levels of monocytes.
3. Results of the hematology did not reveal any treatment-related effects.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
2. Blood chemistry:
In male rat, Ast and Cl were increased, during recovery total cholestrol, PL, glucos and Tp were decreased and Cl were increased in 600 mg/kg/bw/day group.In female, AST, LDH and Ca wee increased and BUN and creatinine level were decreased in 600 mg/kg/be/day group. creatinine level were also decreased in 60 mg/kg/bw/day group female rat. These changes were not observed after the end of recovery period showed reversibility of the change.
3. Blood chemistry examination revealed a slight increase in alanine amino-transferase activity in animals receiving 1000 mg/kg/day when compared with controls.
Urinalysis findings:
not specified
Description (incidence and severity):
2. In male rat treated with 600 mg/kg/day showed red urine sporadically and light brown urine were observed.
3.
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
2.In males treated with 600 mg/kg/day, the absolute organ weight was decreased for thymus, heart, kidneys and epididymis.In the male recovery period, the absolute organ weights for thyroids, heart and liver were decreased.In females treated with 600 mg/kg/day, the absolute organ weight was decreased for brain and thymus, while the organ weight for thyroids, heart, liver, spleen and kidneys increased.In the female recovery period, the absolute organ weights for thyroids, heart and kidneys increased.
3. Organ weight analysis and macroscopic pathology did not reveal any treatment-related effects.
Gross pathological findings:
not specified
Description (incidence and severity):
2. Change attributed in intestinal tracts of male and female of 180 or 600 mg/kg/day groups and in liver of male and female of the 600 mg/kg/day group.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
2. Diffuse hyperplasia in mucosa in the cecum was observed.In rectum, cell debris in crypt was observed in females.In the recovery group, these changes were no longer observed. However in the recovery group, cell infiltration in mucosa was observed in cecum, colon and rectum.In the liver, hypertrophy of perilobular hepatocytes was observed.
3. Histologic evaluations revealed statistically significant increases in histiocytic hyperplasia in the mesenteric lymph nodes in male and female rats receiving 1000 mg/kg/day. A few animals from the 200 mg/kg day dosage group also were affected: however, the group incidence was not statistically significantly different from the control.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Dose descriptor:
NOAEL
Effect level:
> 40 - <= 180 mg/kg bw/day (nominal)
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
urinalysis
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
Critical effects observed:
not specified
Conclusions:
Based on the available results and applying the weight of evidence approach, the NOAEL for the test chemical can be considered to lie between 40-180 mg/kg/day.
Executive summary:

Various studies have been summarized to determine the repeated dose oral toxicity of the test chemical. These include in vivo experimental studies performed on rats for the test chemical. The results are mentioned below:

A Combined Repeated Dose and Reproductive Toxicity Screening Test of the test chemical was performed to evaluate general toxicity and toxicity to reproductive organs and performance and effects on the developing fetus. The study was performed according to OECD 422 Guidelines. A total 96 animals were included in the main group and 20 animals in the recovery groups were used. A total of 12 males and 12 females per group in the main study and 5 males and 5 females per group in the recovery were grouped. A recovery group was included to check the reversibility of the effects that were observed in the main study. The animals that were used in the study were Crl:CD (Sprague Dawley) rats. The animals in all the groups were dosed via the oral route by gavage, using an intubation canula. The doses were prepared in the concentration of 0, 60, 180 and 600 mg/Kg body weight. The vehicle that was used to prepare the doses was purified distilled water. The male animals were dosed for 42 days and the females were dosed from approximately 41 to 53 days (including from 14 days before mating to day 4 lactation period). The recovery group was kept under observation for 28 days to see any reversal of effects that are observed in the main groups. The animals were observed for mortality and clinical signs, Functional Observation Battery, Body weight gain and food consumption, Hematology and Clinical Chemistry parameters. During the observations, at 600 mg/Kg body weight, one male and one female rat were found dead. Before death, the male animals showed fractures of incisors, soft stool and a decrease in the amount of feces. In female rat, staining of lower abdominal fur and a staggering gait was observed. Animals that survived at 600 mg/Kg dose showed presence of red urine sporadically. A light brownish urine was observed during urinanalysis, but the change was transient. Also, at 600 mg/kg male rats showed a tendency of suppressed body weight while the female rats showed signs of increased body weight and food consumption. At 180 mg/Kg body weight, increased food consumption was observed in female rats. In recovery group, at 600 mg/kg, the male animals showed lowering of body weights they were not considered to be of toxicological importance since the same effect was observed in the recovery control group. In clinical chemistry parameters, male rats in 600 mg/Kg showed high values of Aspartate Amino Transfrase (AST) and lower concentration of blood urea nitrogen (BUN), while female rats showed high levels of Aspartate Amino Transferase (AST) and Lactate Dehydrogenase (LDH) and lower levels of blood urea nitrogen (BUN) and creatinine. The females who received dose of 600 mg/Kg showed an increase in the calcium concentration. These changes were found to be inconsistent as the same symptoms were not observed in high dose recovery group. In hematology analysis, no test chemical related adverse effect was observed at any dose levels in main study and the recovery group. The animals were also observed for function observation battery, where all animals were found to be normal and there was no test item related adverse effects in any animals in the main study and the recovery group. The animals were also tested for grip strength analysis wherein there was no change in the animals from all the dose groups when compared to the control group. At 180 mg/Kg and 600 mg/Kg, changes in intestinal tracts in males and female rats. Also, changes in liver morphology were observed at 600 mg/Kg in both males and females. In histopathological examinations, diffused hyperplasia in the mucosa in the cecum was observed, which was assumed to be related to the macroscopic observation in In the rectum, cell debris in crypt was observed in females. In recovery group, all the changes were reverted and thus recovery from these observations was observed. However, cell infiltration in mucosa was observed in cecum, colon and rectum and in liver hypertrophy of perilobular hepatocytes were observed. Thus, from all the above observations, it was concluded that, the observed NOAEL for males and female rats was 180 mg/Kg body weight and LOAEL was considered to be 600 mg/Kg/Day for the test chemical male and female rats.

This result is supported by repeated dose oral toxicity study was performed to determine the toxic nature of the test chemical. The study was performed according to Sixth amendment (79/831/EEC-1979) to the European Community Directive 67/548/EEC Guidelines. The study was performed using male and female Sprague Dawley rats at dose level of 0, 40, 200 and 1000 mg/kg/day. The doses for the main study were based on the dose range finding study. The animals were dosed continuously for 28 days and observed for clinical signs, mortality, body weight and food consumption changes, hematology and clinical chemistry parameters and were subjected to gross pathology and histopathology.No signs of reaction to treatment and no mortality were observed during the treatment period. Food consumption, body weight and food utilization (amount of food consumed per unit of body weight gain) were unaffected by treatment. Results of the hematology did not reveal any treatment-related effects. Blood chemistry examination revealed a slight increase in alanine amino-transferase activity in animals receiving 1000 mg/kg/day when compared with controls. Organ weight analysis and macroscopic pathology did not reveal any treatment-related effects. Histologic evaluations revealed statistically significant increases in histocytic hyperplasia in the mesenteric lymph nodes in male and female rats receiving 1000 mg/kg/day. A few animals from the 200 mg/kg day dosage group also were affected: however, the group incidence was not statistically significantly different from the control. Based on the observations made, No observed adverse effect level (NOAEL) for the test chemical was considered to be 40 mg/Kg/day and the LOAEL can be considered to be 200 mg/kg/day

Based on the available results and applying the weight of evidence approach, the NOAEL for the test chemical can be considered to lie between 40-180 mg/kg/day.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
200 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
Data is Klimisch 2 and from authoritative database

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
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:
short-term repeated dose toxicity: dermal
Data waiving:
study scientifically not necessary / other information available
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

Various studies have been summarized to determine the repeated dose oral toxicity of the test chemical. These include in vivo experimental studies performed on rats for the test chemical. The results are mentioned below:

A Combined Repeated Dose and Reproductive Toxicity Screening Test of the test chemical was performed to evaluate general toxicity and toxicity to reproductive organs and performance and effects on the developing fetus. The study was performed according to OECD 422 Guidelines. A total 96 animals were included in the main group and 20 animals in the recovery groups were used. A total of 12 males and 12 females per group in the main study and 5 males and 5 females per group in the recovery were grouped. A recovery group was included to check the reversibility of the effects that were observed in the main study. The animals that were used in the study were Crl:CD (Sprague Dawley) rats. The animals in all the groups were dosed via the oral route by gavage, using an intubation canula. The doses were prepared in the concentration of 0, 60, 180 and 600 mg/Kg body weight. The vehicle that was used to prepare the doses was purified distilled water. The male animals were dosed for 42 days and the females were dosed from approximately 41 to 53 days (including from 14 days before mating to day 4 lactation period). The recovery group was kept under observation for 28 days to see any reversal of effects that are observed in the main groups. The animals were observed for mortality and clinical signs, Functional Observation Battery, Body weight gain and food consumption, Hematology and Clinical Chemistry parameters. During the observations, at 600 mg/Kg body weight, one male and one female rat were found dead. Before death, the male animals showed fractures of incisors, soft stool and a decrease in the amount of feces. In female rat, staining of lower abdominal fur and a staggering gait was observed. Animals that survived at 600 mg/Kg dose showed presence of red urine sporadically. A light brownish urine was observed during urinanalysis, but the change was transient. Also, at 600 mg/kg male rats showed a tendency of suppressed body weight while the female rats showed signs of increased body weight and food consumption. At 180 mg/Kg body weight, increased food consumption was observed in female rats. In recovery group, at 600 mg/kg, the male animals showed lowering of body weights they were not considered to be of toxicological importance since the same effect was observed in the recovery control group. In clinical chemistry parameters, male rats in 600 mg/Kg showed high values of Aspartate Amino Transfrase (AST) and lower concentration of blood urea nitrogen (BUN), while female rats showed high levels of Aspartate Amino Transferase (AST) and Lactate Dehydrogenase (LDH) and lower levels of blood urea nitrogen (BUN) and creatinine. The females who received dose of 600 mg/Kg showed an increase in the calcium concentration. These changes were found to be inconsistent as the same symptoms were not observed in high dose recovery group. In hematology analysis, no test chemical related adverse effect was observed at any dose levels in main study and the recovery group. The animals were also observed for function observation battery, where all animals were found to be normal and there was no test item related adverse effects in any animals in the main study and the recovery group. The animals were also tested for grip strength analysis wherein there was no change in the animals from all the dose groups when compared to the control group. At 180 mg/Kg and 600 mg/Kg, changes in intestinal tracts in males and female rats. Also, changes in liver morphology were observed at 600 mg/Kg in both males and females. In histopathological examinations, diffused hyperplasia in the mucosa in the cecum was observed, which was assumed to be related to the macroscopic observation in In the rectum, cell debris in crypt was observed in females. In recovery group, all the changes were reverted and thus recovery from these observations was observed. However, cell infiltration in mucosa was observed in cecum, colon and rectum and in liver hypertrophy of perilobular hepatocytes were observed. Thus, from all the above observations, it was concluded that, the observed NOAEL for males and female rats was 180 mg/Kg body weight and LOAEL was considered to be 600 mg/Kg/Day for the test chemical male and female rats.

This result is supported by repeated dose oral toxicity study was performed to determine the toxic nature of the test chemical. The study was performed according to Sixth amendment (79/831/EEC-1979) to the European Community Directive 67/548/EEC Guidelines. The study was performed using male and female Sprague Dawley rats at dose level of 0, 40, 200 and 1000 mg/kg/day. The doses for the main study were based on the dose range finding study. The animals were dosed continuously for 28 days and observed for clinical signs, mortality, body weight and food consumption changes, hematology and clinical chemistry parameters and were subjected to gross pathology and histopathology.No signs of reaction to treatment and no mortality were observed during the treatment period. Food consumption, body weight and food utilization (amount of food consumed per unit of body weight gain) were unaffected by treatment. Results of the hematology did not reveal any treatment-related effects. Blood chemistry examination revealed a slight increase in alanine amino-transferase activity in animals receiving 1000 mg/kg/day when compared with controls. Organ weight analysis and macroscopic pathology did not reveal any treatment-related effects. Histologic evaluations revealed statistically significant increases in histocytic hyperplasia in the mesenteric lymph nodes in male and female rats receiving 1000 mg/kg/day. A few animals from the 200 mg/kg day dosage group also were affected: however, the group incidence was not statistically significantly different from the control. Based on the observations made, No observed adverse effect level (NOAEL) for the test chemical was considered to be 40 mg/Kg/day and the LOAEL can be considered to be 200 mg/kg/day

Based on the available results and applying the weight of evidence approach, the NOAEL for the test chemical can be considered to lie between 40-180 mg/kg/day.

Repeated dose toxicity: Inhalation

A short term toxicity study does not need to be conducted because exposure of humans via inhalation route is highly unlikely based on the thorough and rigorous exposure assessment. The test chemical has very low vapor pressure of 2.61E-10 Pa which is equivalent to 1.96E-12 mm Hg at 25 ° C, so the potential for the generation of inhalable vapour is low. Also the normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be unlikely to occur, and therefore this end point was considered for waiver

Repeated dose toxicity: Dermal

A short term toxicity study does not need to be conducted because exposure of humans via inhalation route is highly unlikely based on the thorough and rigorous exposure assessment. In accordance with ANNEX VII Colum 2 of the REACH regulation, the study need not be conducted if the substance is a strong acid (pH<=2.0) or strong base (pH=> 11.5). The experimental pH of the test chemical is 14. Hence, based on the high pH of the test chemical, the repeated dermal toxicity study was considered for waiver.

Justification for classification or non-classification

Based on the data available for the test chemical N,N,N-tributylbutan-1-aminium hydroxide (2052-49-5) not likely to exhibit toxic nature upon repeated exposure by oral, inhalation and dermal route of exposure and hence is not likely to classify as per the criteria mentioned in CLP regulation.