Registration Dossier

Administrative data

Description of key information

Oral:

The studies available are feeding studies in rat and mice (one drinking water study in mice). Mice seem to be less susceptible to the effects of the test substance than rats. Effects are severe decrease of body weight with effects in the liver. The no effect level is 2% in the diet of rats. In the reports available different conversion methods are used to calculate the actual test substance intake. This may have been done by actual measurement of food intake, but is not clearly described in the publications. Therefore a calculation was done based on the factors as described in the WHO report (EHS Environmental Health Criteria 240). This leads to a NOAEL of 1000 mg/kg bw based on the 2% dietary level in rats.

In a chronic toxicity/carcinogenicity study in rats (Sodemoto, 1979) no signs of toxicity were reported at 2% the test substance in feed (stated to be equivalent to 1000 mg/kg bw). In mice (Toth, 1984) no effects were reported at 2% the test substance in drinking water (119-124 mg/animal/day).

Dermal: An repeated dermal toxicity study with rabbit (Marroquin, 1981) which run on analog substance Benzoic acid (65-85-0) is available which is key study.The NOAEL was > 2500 mg/kg bodyweight.

Inhalation: A study is available with a NOAEL of the key study of 250 mg/m3 (Benson, 1981). These findings were primarily attributed to the physico-chemical properties of these fine low-solubility particles.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Limited study summary. Study predates approved guidelines, minimal data provided on study design and results, no GLP.
Qualifier:
no guideline followed
Principles of method if other than guideline:
Study predates approved guidelines.
The chronic toxicity study of the test substance was examined in Fischer 344 rats. The test substance was administered in the diet for 18 to 24 months at two dose levels.
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
Fischer 344
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Provided by Japan CLEA Co. Ltd.
- Age at study initiation: 4-5 weeks old
- Weight at study initiation: 110-150 g
- Housing: Five in each cage
- Diet (e.g. ad libitum): CE-2 pellets, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-23℃
- Humidity (%): 55%

No additional data
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
None stated
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
18-24 months
Frequency of treatment:
Daily
Remarks:
Doses / Concentrations:
1 or 2%
Basis:
nominal in diet
No. of animals per sex per dose:
Test group: 50 males and 52 females per dose
Control group: 25 males and 43 females
Control animals:
yes, plain diet
Details on study design:
Both male and female rats were divided into three groups and maintained on the diet containing 2 or 1% of the test substance or basal diet for 18 to 24 months. The animals were maintained on a free choice diet, but amount of diet was adequately controlled to avoid excess. Tap water was freely offered to all animals. The rats were weighed weekly, and food intake was measured daily.
Positive control:
None stated
Observations and examinations performed and frequency:
Mortality, growth, food intake, behavior and general status were observed in the experimental period.
Sacrifice and pathology:
In the middle of the experimental period, several animals from each groups selected at random were sacrificed for morphologic examination. All surviving animals were killed between 18 to 25 months. For morphologic examination, autopsy was carried out in all animals and paraffin sections of various organs were prepared and stained (hematoxylin and eosin).
Other examinations:
None stated
Statistics:
T-test
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
no effects observed
Details on results:
Clinical signs and mortality:
During the first 16 months of the experimental period, the average mortality rate of all animals was 14.5%. Except for myeloproliferative disorder developed in one female control rat, all the dead animals showed pneumonia with abscess. Around 100 rats including those of the control groups died after 16 months of hemorrhagic pneumonia with edema which was probably induced by mixed infection of sialodacryoadenitis and mycoplasma. However, no differences in mortality rates between treated and control groups were observed throughout the experimental course.

Body weight gain and food intake:
Nor were there any significant differences in growth and food intake between groups of rats.

Histopathology:
Most developed tumours were benign, such as interstitial cell tumour of the testes and endometrial polyp of the uterus. However, a small number of malignant neoplasms including leukemia was also observed. Differences in numbers of tumor-bearing animals and time-to-tumor development between control and treated gorups were not significant (t-test). No specific toxic effect induced by the test substance was observed.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No findings of toxicological significance at 20000 mg/kg diet (equivalent to 1000 mg/kg bw)..
Critical effects observed:
not specified
Conclusions:
From the results of this study, it may be concluded that the test substance was not toxic for male and female Fischer 344 rats after 18 to 24-month dietary exposure.
Executive summary:

The chronic toxicity study of the test substance was examined in Fischer 344 rats. The test substance was administered in the diet for 18 to 24 months at two dose levels.

No adverse clinical signs directly attributable to the compound were observed in treated animals. Differences in the average body weight, and mortality rates between treated and control groups were negligible. The results of the statistical test for dose-related trends were significant (p<0.05). Although a variety of tumors occurred among test and control rats of each sex, tumors appearing in treated rats were similar in type and number to those in controls. It was concluded that no evidence of carcinogenicity in rats from the test substance was demonstrated.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
chronic
Species:
rat
Quality of whole database:
2 (reliable with restrictions)

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: inhalation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
From 14 March to 11 April 1980
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study run to a method comparable with current guidelines and GLP.
Justification for type of information:
See Read Across Justification document in Section 13.2 of IUCLID
Reason / purpose:
read-across source
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 412 (Subacute Inhalation Toxicity: 28-Day Study)
Principles of method if other than guideline:
This study consisted of four groups, each containing ten male and ten female albino rats. One group served as an air exposed control while the other three were exposed to one of three different dust concentrations of the test substance. Exposures were for 6 hours per day five days per week for four consecutive weeks. At various times during the study body weights and observations for pharmacotoxic signs were recorded. After four weeks of exposure various serum biochemical, hematologic, organ weight and histopathologic evaluations were conducted.
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories, Inc.
- Age at study initiation: At the time of receipt the rats were 42 days of age.
- Weight at study initiation: males 194-201 g, females 148-163 g
- Housing: caged individually
- Diet: ad libitum for non-exposure hours
- Water: tap water, ad libitum for non-exposure hours
- Acclimation period: yes

ENVIRONMENTAL CONDITIONS
- Temperature (°F): 71.6-74.7°F
- Humidity (%): 41.0-51.0%
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12 hour photoperiod

IN-LIFE DATES: From: 14 March 1980 To: 11 April 1980
Route of administration:
inhalation: dust
Type of inhalation exposure:
whole body
Vehicle:
air
Remarks on MMAD:
MMAD / GSD: A mean equivalent aerodynamic diameter of 4.7 µm was defined.
Group 2, EAD of 4.6 µm, ag of 3.1
Group 3, EAD of 4.4 µm, ag of 2.1
Group 4, EAD of 5.2 µm, ag of 2.1
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Exposures were conducted in 1 cubic meter glass and stainless steel exposure chambers.
- Method of holding animals in test chamber:
- Source and rate of air: Air for chamber ventilation was supplied from an HVAC system separate from general laboratory systems.
- Method of conditioning air:
- System of generating particulates/aerosols: The test material was ground in an Oster@ blender in order to produce a more respirable particle. The blender jar was filled approximately one third full with the test substance and run at high speed for one to two minutes. The ground compound was separated from the remaining flakes by using a flour sifter. The flakes were returned to the blender jar and reground. The procedure was continued until enough material for two exposure days had been prepared.
- Temperature, humidity, pressure in air chamber:
- Air flow rate: Chamber air flow rate varied between one 145 and 310 liters per minute depending on desired exposure concentrations.
- Air change rate:
- Method of particle size determination: The particle size distribution of the test material in a sample of the chamber atmosphere was determined using an Andersen 8 stage cascade impactor.
- Treatment of exhaust air:

TEST ATMOSPHERE
- Brief description of analytical method used: Nominal concentrations were determined by weighing the amount of test material placed in the generator reservoir prior to exposure and then again after exposure. The difference in weight was the nominal concentration. Actual exposure concentrations were determined by standard gravimetric techniques. Exposure atmospheres were drawn through pre-weighed glass fiber filters at a known flowrate for a known time. The filters were reweighed, and the difference was divided by the total air volume to yield the actual concentrations.
- Samples taken from breathing zone: yes/no

VEHICLE (if applicable)
- Justification for use and choice of vehicle:
- Composition of vehicle:
- Type and concentration of dispersant aid (if powder):
- Concentration of test material in vehicle:
- Lot/batch no. of vehicle (if required):
- Purity of vehicle:
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Both nominal and analytical exposure concentrations were determined for this study. Nominal concentrations were determined by weighing the amount of test material placed in the generator reservoir prior to exposure and then again after exposure. The difference in weight was divided by the total air passed through the chamber during the exposure period resulting in the nominal concentration.
Actual exposure concentrations were determined by standard gravimetric techniques. Exposure atmospheres were drawn through pre-weighed glass fiber filters at a known flowrate for a known time. The filters were reweighed and the difference was divided by the total air volume to yield the actual concentrations.

Group 2, desired concentration 0.02 mg/L, nominal concentration 0.31 mg/L, actual concentration 0.025 mg/L
Group 3, desired concentration 0.2 mg/L, nominal concentration 2.2 mg/L, actual concentration 0.25 mg/L
Group 4, desired concentration 2.0 mg/L, nominal concentration 16.5 mg/L, actual concentration 1.2 mg/L

Duration of treatment / exposure:
4 weeks
Frequency of treatment:
Animals were exposed for 6 hours per day, 5 days per week for 4 weeks.
Remarks:
Doses / Concentrations:
25, 250, 1200 mg/m3
Basis:
analytical conc.
Remarks:
Doses / Concentrations:
310, 2200, 16500 mg/m3
Basis:
nominal conc.
Remarks:
Doses / Concentrations:
20, 200, 2000 mg/m3
Basis:
other: Desired concentration
No. of animals per sex per dose:
10 male and 10 female per dose
Control animals:
yes, concurrent no treatment
Details on study design:
- Dose selection rationale:
- Rationale for animal assignment (if not random): The animals were randomized into the four groups based on body weight.
- Rationale for selecting satellite groups:
- Post-exposure recovery period in satellite groups:
- Section schedule rationale (if not random):
Positive control:
None stated
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: once each week
- Cage side observations checked in table [No.?] were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Twice each day prior to exposure and again after each exposure

BODY WEIGHT: Yes
- Time schedule for examinations: prior to exposure and after 1, 2, 3 and 4 weeks of exposure

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data

WATER CONSUMPTION: No data

OPHTHALMOSCOPIC EXAMINATION: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood: after four weeks of exposure
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: all survivors
- Parameters: Hematocrit, Hemoglobin Concentration, Erythrocyte count, Leucocyte Count (total and differential), Platelet Count, MCV, MCH, MCHC


CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: after four weeks of exposure
- Animals fasted: No data
- How many animals: all survivors
- Parameters: Blood Urea Nitrogen, Serum Alkaline Phosphatase, Serum Glutamic Pyruvic Transaminase, Serum Glutamic Oxaloacetic Transaminase, Blood Glucose

URINALYSIS: No data

NEUROBEHAVIOURAL EXAMINATION: No data
Sacrifice and pathology:
GROSS PATHOLOGY: Yes, heart, kidney, lungs/trachea, brain, liver, spleen
HISTOPATHOLOGY: Yes, adrenal, nasal turbinate, brain, pancreas, colon, pituitary, esophagus, prostate/uterus, eye with optic nerve, submaxillary salivary gland, testis (both), ovary, jejunum, Harderian glands, spleen, heart, sternum (bone marrow), kidney, stomach, liver, thymus, lungs (5 lobes), thyroid/parathyroid, bronchial lymph node, urinary bladder, mammary gland
Other examinations:
None stated
Statistics:
Body weight data, clinical chemistry data, hematology data and organ weight data are presented on both an individual animal basis and as a group mean and standard deviation summary. Not all parameters were statistically evaluated at all intervals. Some parameters at some intervals were evaluated statistically after a review of the data indicated that a possible exposure related effect may have been present. All statistical analyses compared the treatment groups with the control group by sex.
Body weights (weeks 1, 2, 3 and 4), hematological parameters (week 4) and absolute and relative organ weights (terminal sacrifice) were compared by analysis of variance (one-way classification), Bartlett's test for homogeneity of variances and the appropriate t-test (for equal or unequal variances) as described by Steel and Torrie using Dunnett's multiple comparison tables to judge significance of differences.
Clinical signs:
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
not specified
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
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
Reddisch discharge around the nares was seen in the mid and high dose groups. Two high dose animals died during the study.

BODY WEIGHT AND WEIGHT GAIN
High dose males and females showed a decreased body weight gain.

HAEMATOLOGY
A statistically significant decrease in platelets was seen in high dose males and females

CLINICAL CHEMISTRY
Random statistical differences were observed in the biochemical data. These differences were not considered to be exposure related nor of toxicological significance.

ORGAN WEIGHTS
Changes in absolute and relative weights of liver, kidneys and lungs were noted in high dose males or females.

GROSS PATHOLOGY
No changes in gross pathology were noted.

HISTOPATHOLOGY: NON-NEOPLASTIC
Compound related microscopic lesions were confined to the lung. An increased incidence and intensity of interstitial inflammatory cell infiltrate interstitial fibrosis was noted in low, mid and high dose animals.
Dose descriptor:
NOAEC
Effect level:
<= 25 mg/m³ air
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Pulmonary fibrosis and inflammatory cell infiltrate at lowest dose level
Dose descriptor:
NOAEL
Remarks:
systemic
Effect level:
250 mg/m³ air
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Only very slight organ weight and body weight decreases noted.
Critical effects observed:
not specified
Conclusions:
All test concentrations induced local effects: nasal redness and discharge and pulmonary fibrosis and inflammatory cell infiltrates in the lungs that can be related to the ittitant properties of the test substance. No systemic effects were noted at 25 mg/m3. At 250 mg/m3 a very slight decrease in absolute kidney weighth was seen in females only. At the highest dose level mortality, decresed body weights as well as decreased liver kidney and lung weights were reported. No histopathological findings except in the lungs were reported . No systemic effects were noted at this dose level. At 250 mg/m3 a very slight decrease in absolute kidney weighth was seen in females only. At the highest dose level mortality, decresed body weights as well as decreased liver kidney and lung weights were reported. No histopathological findings except in the lungs were reported .
Executive summary:

This study consisted of four groups, each containing ten male and ten female albino rats. One group served as an air exposed control while the other three were exposed to one of three different dust concentrations of the test substance. Exposures were for 6 hours per day five days per week for four consecutive weeks. At various times during the study body weights and observations for pharmacotoxic signs were recorded. After four weeks of exposure various serum biochemical, hematologic, organ weight and histopathologic evaluations were conducted.

All test concentrations induced local effects: nasal redness and discharge and pulmonary fibrosis and inflammatory cell infiltrates in the lungs that can be related to the irritant properties of the test substance. No systemic effects were noted at 25 mg/m3. At 250 mg/m3 a very slight decrease in absolute kidney weighth was seen in females only (body weight was also slightly lower (not significantly) than in control females). At the highest dose level mortality, decreased body weights as well as decreased liver kidney and lung weights were reported. No histopathological findings except in the lungs were reported . The NOAEL for local effects is < 25 mg/m3; The NOAEL for systemic effects is 250 mg/m3.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEC
250 mg/m³
Study duration:
subacute
Species:
rat
Quality of whole database:
1 (reliable without restriction)

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 14 November to 5 December 1979
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study run to a reliable method and to GLP.
Justification for type of information:
See Read Across Justification document in Section 13.2 of IUCLID
Reason / purpose:
read-across source
Reason / purpose:
read-across: supporting information
Qualifier:
according to
Guideline:
EPA OPP 82-2 (Repeated Dose Dermal Toxicity -21/28 Days)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Langshaw Farms, Augusta, Michigan
- Age at study initiation:
- Weight at study initiation: 2127 to 2891 grams
- Fasting period before study:
- Housing: Individually housed in hanging wire-mesh cages.
- Diet (e.g. ad libitum): Purina Certified Rabbit Chow, ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 13 days

ENVIRONMENTAL CONDITIONS
- Temperature (°F): 70-76 °F
- Humidity (%): 44-57%
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark

IN-LIFE DATES: From: 14 November 1979 To: 5 December 1979
Type of coverage:
semiocclusive
Vehicle:
physiological saline
Remarks:
0.9%
Details on exposure:
TEST SITE
- Area of exposure: 100 cm2
- % coverage: 23%, 35% and 100%
- Type of wrap if used: Each site was covered with a 10 x 10-centimeter gauze pad with adhesive tape borders. The entire area was then wrapped with gauze bandaging and Elastoplast tape.
- Time intervals for shavings or clipplings: twice each week prior to test article administration during the three week study period

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The test areas were washed with tepid IRDC tap water and dried with disposable paper towels.
- Time after start of exposure: 6 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100, 500 and 2500 mg/kg bw
- For solids, paste formed: no

VEHICLE
not applicable

USE OF RESTRAINERS FOR PREVENTING INGESTION: no
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
6 hours, test substance moistened with physiological saline
Frequency of treatment:
Once a day, 5 days per week, for 3 consecutive weeks
Remarks:
Doses / Concentrations:
100, 500, 2500 mg/kg
Basis:
nominal per unit body weight
No. of animals per sex per dose:
4 males and 4 females per dose
Control animals:
yes
Details on study design:
None stated
Positive control:
None stated
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: once daily during the 21-day study period
- Cage side observations checked in table [No.?] were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once daily

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: once daily

BODY WEIGHT: Yes
- Time schedule for examinations: Body weights were obtained during the pretest period, prior to study initiation and at weekly intervals during the 21-day study period

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data

WATER CONSUMPTION: No data

OPHTHALMOSCOPIC EXAMINATION: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood: once during the pretest period and at 21 days of study
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: all animals
- Parameters: hematocrit, hemoglobin, erythrocyte count, total leucocyte count, differential leucocyte count, platelets, MCH, MCV and MCHC

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Once during the pretest period and at 21 days of study.
- Animals fasted: Yes / No / No data
- How many animals: all animals
- Parameters:glucose, blood urea nitrogen, lkaline phosphatase, serum glutamic pyruvic transaminase, and serum glutamic oxalacetic transaminase

URINALYSIS: No data

NEUROBEHAVIOURAL EXAMINATION: No data

OTHER:
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
masses and lesions, untreated skin, treated skin, kidney, lungs, pituitary, sciatic nerve, muscle, spleen, urinary bladder, thymus, lumbar spinal cord, pancreas, stomach, duodenum, testes, thyroid, femur, colon, mesenteric lymph nodes, mediastinal lymph nodes, ovaries, parathyroid, liver, gall bladder, adrenals, heart, brain
HISTOPATHOLOGY: Yes
spleen, pancreas, stomach, duodenum, colon, lymph nodes, urinary bladder, heart, lung, brain (3 sections), treated skin (3 sections), untreated skin (3 sections), liver, kidney, gall bladder, adrenals, spinal cord, gonads, nerve (with muscle), bone marrow, thyroid (parathyroid), pituitary, thymus
Other examinations:
None stated
Statistics:
All statistical analyses compared the treatment groups with the control group, by sex.
Body weights (day 21), hematological and biochemical parameters (pretest and day 21) and absolute and relative organ weights (terminal sacrifice) were compared by analysis of variance (one-way classification), Bartlett’s test for homogeneity of variances and the appropriate t-test (for equal or unequal variances) as described by Steel and Torrie using Dunnett’s multiple comparison tables to judge significance of differences.
Clinical signs:
no effects observed
Dermal irritation:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
not specified
Details on results:
CLINICAL SIGNS AND MORTALITY
In the control group, a few rabbits exhibited signs of anorexia and purulent nasal discharge. One rabbit also exhibited diarrhea.
In the test groups, a few rabbits exhibited signs of anorexia, purulent nasal discharge, clear nasal discharge, soft stool, diarrhea and ocular discharge.
None of the rabbits died during the 21-day study period.

DERMAL IRRITATION
Very slight dermal irritation was noted for one rabbit at the 2500 mg/kg dosage level.

BODY WEIGHT AND WEIGHT GAIN
There were no statistical differences found between the control and test groups.

HAEMATOLOGY
There were no treatment-related differences noted in the hematology data in this study.

CLINICAL CHEMISTRY
Although there was one statistically significant difference noted, this was not considered physiologically meaningful or treatment-related. The mid-dose female SGOT mean was significantly lower than the control mean, however, there were only four females in each group and the difference was only 5 IU/1. Therefore the statistically significant difference observed does not have any biological significance.

ORGAN WEIGHTS
The comparison of the control and treated group means for body and organ weights did not show any important differences.

GROSS PATHOLOGY
The skin from the application site was described as erythematous in one, two and seven animals of groups I, III and IV, respectively. The lungs presented foci or areas of congestion that were considered unrelated to the application of the test compound.

HISTOPATHOLOGY: NON-NEOPLASTIC
The skin from the application site presented mainly infiltration of inflammatory cells in the dermis with slight severity in both control and treated groups. Slight hyperkeratosis was observed in three males and four females in group IV. These lesions were very slight and represent a minimal reaction to the test compound.
The foci of myocarditis found in some treated animals were considered unrelated to the application of the test compound since they are observed from time to time in control animals.
Dose descriptor:
NOAEL
Effect level:
> 2 500 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: One female in the 2500 mg/kg test group exhibited dermal irritation during the 21-day study period.
Critical effects observed:
not specified
Conclusions:
The NOAEL was considered to be > 2500 mg/kg. Very slight dermal irritation was noted for one rabbit at this dosage level. No compound related changes were seen in general behavior and appearance, body weight, clinical laboratory tests, organ weights or survival.
Executive summary:

In a dermal study in New Zealand White rabbits, the test substance was applied 5 days a week for 3 weeks at dosage levels of 100, 500 and 2500 mg/kg. Four male and four female rabbits were used in each treated group and in a control group. 0.9% physiological saline was used as the control substane. The rabbits were observed daily for signs of dermal irritation and changes in general behavior and appearance. Individual body weights were recorded weekly Hematologic and biochemical studies were conducted once in the pretest period and at 21 days of study.

The NOAEL was considered to be > 2500 mg/kg. Very slight dermal irritation was noted for one rabbit at this dosage level. No compound related changes were seen in general behavior and appearance, body weight, clinical laboratory tests, organ weights or survival.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
2 500 mg/kg bw/day
Study duration:
subacute
Species:
rabbit
Quality of whole database:
1 (reliable without restriction)

Additional information

Oral:

A short-term dietary study (10 days) in rats and mice (Fujitani, 1993) showed a NOAEL of 905 mg/kg bw in rats and of 3571 mg/kg bw in mice. Effects included decreased body weight and increased (rel) liver weights with enlarged (glossy) hepatocytes and eosininophilic foci around the protal vein. In high dosed decreased kidney weights were reported without histopathological changes. In addition, in a 35 day study in rats (Sodemoto, 1979) animals at the highest dose groups died. Symptoms in decendents were severe decrease in body weight. Morpholical effects were limited to atrophy of the spleen and lymphnodes at 4 and 8% in diet. At the NOAEL 2% in diet no morphological changes were reported. In a limited reported 90-day study in rats (Weil, 1953) a NOAEL of 2620 mg/kg bw was established based on reduced body weight gain, increased relative liver and kidney weights and pathological changes in liver and kidneys at 6290 mg/kg bw/day.

The overall NOAEL for repeated dose toxicity is based on a chronic toxicity study and is set at 1000 mg/kg bw/day (Sodemoto, 1979).

Dermal:

A 21-day dermal toxicity study with rabbit (Marroquin, 1981) which run on analog substance Benzoic acid (65-85-0) is available. No effects were found at the highest dose level, thus the NOAEL was > 2500 mg/kg bodyweight.

Inhalation:

A repeated inhalation toxicity study with rat (Benson, 1981) which was run on an analog substance Benzoic acid (65-85-0) is available. All test concentrations induced local effects: nasal redness and discharge and pulmonary fibrosis and inflammatory cell infiltrates in the lungs that can be related to the irritant properties of the test substance.  Hence these are not taken into account in relation to classification.

No systemic effects were noted at 25 mg/m3. At 250 mg/m3 a very slight decrease in absolute kidney weight was seen in females only (body weight was also slightly lower (not significantly) than in control females). At the highest dose level mortality, decreased body weights as well as decreased liver kidney and lung weights were reported. No histopathological findings except in the lungs were reported. The NOAEL for local effects is < 25 mg/m3; The NOAEL for systemic effects is 250 mg/m3. At 1,200 mg/m3 significant adverse toxicological effects were observed.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:

This study was read-across from benzoic acid. It was carried out with rabbits according to a reliable method.

Justification for selection of repeated dose toxicity inhalation - systemic effects endpoint:

Only one read-across study from benzoic acid is available.

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:

Only one read-across study from benzoic acid is available.

Repeated dose toxicity: inhalation - systemic effects (target organ) respiratory: lung

Justification for classification or non-classification

Oral: NOAEL > 1000 mg/kg bw in a chronic toxicity study > 90 days.

Therefore in accordance with Regulation (EC) No. 1272/2008 Table 3.9.2 and 3.9.3 the substance is not classified for the repeated oral toxicity endpoint.

Dermal: A dermal value of 833 mg/kg bw (which is above 200 mg/kg bw) does not lead to the substance being classified.

Inhalation: Classification of STOT RE comes from the consideration of the dose/concentration which has been shown to produce significant health effects. In Bensen 1981 significant systemic toxic effects were seen to occur at 400 mg/m3 (28 day result: 1200 mg/m3). Local effects are not taken into consideration as these are considered to be related to the irritant nature of the substance and not associated with substance toxicity itself. This then leads to the substance not being classified for inhalation (STOT RE effect >200 mg/m3).