Registration Dossier

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Administrative data

Description of key information

Sanders (2011), OECD 420: Acute Oral tox. Category 4


Khalepo (1968), Acute oral toxicity rat: Inconclusive 


Khalepo (1968), Acute Oral toxicity mouse: Inconclusive


Khalepo (1968), Acute Oral toxicity rabbit: Inconclusive


Khalepo (1968), Acute Inhalation toxicity rat: Inconclusive


Khalepo (1968), Acute Inhalation toxicity mouse: Inconclusive


Khalepo (1968), Acute dermal toxicity rabbit: Inconclusive 


The in vivo acute toxicity studies conduced by Khalepo 1968 were not conclusive for classification therefore the classification of the test substance required the use of an additional in vivo study. The results from the in vivo OECD TG 420 were sufficient to classify the test item to be an acute tox oral Category 4 classification in accordance to the CLP Regulation (EC) No 1272/2008. 

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: oral
Type of information:
other: Published paper
Adequacy of study:
disregarded due to major methodological deficiencies
Study period:
Not stated
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Remarks:
The literature paper does not provide sufficient information on the study.
Qualifier:
no guideline available
Principles of method if other than guideline:
No guideline indicated.
GLP compliance:
no
Test type:
other: No test type indicated
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: Not stated
- Purity, including information on contaminants, isomers, etc.: Not stated

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Not stated
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: Not stated
- Stability in the medium, i.e. sensitivity of the test material to hydrolysis and/or photolysis: Not stated
- Solubility and stability of the test material in the solvent/vehicle and the exposure medium: Not stated
- Reactivity of the test material with the incubation material used (e.g. plastic ware): Not stated

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing (e.g. warming, grinding): Not stated
- Preliminary purification step (if any): Not stated
- Final concentration of a dissolved solid, stock liquid or gel: Emulsion
Species:
rat
Strain:
not specified
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Rationale for alternative/additional species to rat: Not stated
- Source: Not stated
- Females (if applicable) nulliparous and non-pregnant: Not stated
- Rationale for use of males: Not stated
- Age at study initiation: Not stated
- Weight at study initiation: 180 - 220g
- Fasting period before study: Not stated
- Housing: Not stated
- Historical data:Not stated
- Diet (e.g. ad libitum): Not stated
- Water (e.g. ad libitum): Not stated
- Acclimation period: Not stated
- Microbiological status :Not stated
- Method of randomisation in assigning animals to test and control groups :Not stated

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Not stated
- Humidity (%): Not stated
- Air changes (per hr): Not stated
- Photoperiod (hrs dark / hrs light): Not stated
Route of administration:
oral: gavage
Vehicle:
not specified
Details on oral exposure:
VEHICLE
- Concentration in vehicle: Not stated
- Amount of vehicle (if gavage): Not stated
- Justification for choice of vehicle: Not stated
- Lot/batch no. (if required): Not stated
- Purity: Not stated

MAXIMUM DOSE VOLUME APPLIED: 0.2ml

DOSAGE PREPARATION (if unusual):Material prepared as an emulsion

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Not stated
Doses:
Not stated
No. of animals per sex per dose:
6 animals were used
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: Two weeks
- Frequency of observations and weighing: observation at 40 - 60 mins following admistration
- Necropsy of survivors performed: Yes
- Clinical signs including body weight : Clinical signs were reported within the results, but a comprehensive list of all monitored not reported
- Other examinations performed: clinical signs, body weight, organ weights, histopathology, other: Not stated
Statistics:
Statistics was processed based on probe-analysis Litchfield and Wilcoxon methods modified by Rot.
Key result
Sex:
female
Dose descriptor:
LD50
Remarks:
(reported in the source paper as DL50)
Effect level:
720 mg/kg bw
Based on:
other: Test material in emulsion form
Key result
Sex:
male
Dose descriptor:
LD50
Remarks:
(reported in the source paper as DL50)
Effect level:
480 mg/kg bw
Based on:
not specified
Remarks:
Based on test material emulsion
Mortality:
It is indicated that animals died during the study but no additional information is provided.
Clinical signs:
Administration of m-ABTF showed the presence of methemoglobin formers, in 40 - 60 minutes after administration of the test substance, cyanosis and adynamia was observed followed by decreased breathing and reduced reflexes.
Body weight:
Not stated
Gross pathology:
Morphological examinations of the internal organs state moderate macrovesicular hepatic steatosis/fatty liver and protein degeneration of the liver cell. Internal organs hyperemia was also observed.
Other findings:
- Organ weights: Not stated
- Histopathology: Not stated
- Potential target organs: Nervous system
- Other observations: An increase in oxygen consumption was noted with reduced rectal temperatures

Table 2: m-ABTF toxicity when introduced to the stomach





























Dose*Species Mg/kgMmol/kg
DL16Rats3001.86
DL50Rats 4802.96
DL84Rats7654.74

* Statistics was processed based on probe-analysis Litchfield and Wilcox methods modified by Rot (M.L. Belenky 1959)


 


Table 3: Average lethal dose of m-ABTF when introduced to stomch of femal and male rats. 


 





















Species gender



DL50 in mg/kg



m-ABTF


 



Male



480



Female



720


Interpretation of results:
study cannot be used for classification
Conclusions:
Under the limited conditions of this study outlined in the published paper the test material has an LD50 for males of 480 mg/kg and 720 mg/kg for females.
Executive summary:

The published paper pre-dates the existence of nationally or internationally recognised test methods. No specific details on the test method were provided. The limited information presented indicates that 0.2ml m-Aminobenzotrifluoride (m-ABTF, a synonym for alpha,alpha,alpha-trifluoro-m-toluidine) in emulsion form was introduced to the stomach of 6 rats per dose group (160 rats in total) in a single exposure. Animal death and survival was recorded over a 2-week period. 40 - 60 minutes after the administration of m-ABTF cyanosis and adynamia was observed followed by decreased breathing and reflexes. Morphological examinations of the animals indicated the manifestation of methemoglobin formation, with the internal organs exhibiting moderate macrovesicular hepatic steatosis/fatty liver and protein degeneration of the liver cells, alongside hyperaemia of the internal organs. The LD50 for males was considered to be 480 mg/kg bw and 720mg/kg bw for females. Results presented within Table 2 are not indicated as sex specific and do not offer any additional information to the test outcome. Due to the limited information available on the study conducted this data is not considered reliable for use in the classification of the substance according to CLP Regulation (EC) No 1272/2008.

Endpoint:
acute toxicity: oral
Type of information:
other: Published paper
Adequacy of study:
disregarded due to major methodological deficiencies
Study period:
Not stated
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Remarks:
The literature paper does not provide sufficient information on the study
Qualifier:
no guideline available
Principles of method if other than guideline:
Not stated
GLP compliance:
no
Test type:
other: Not stated
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier) and lot/batch number of test material: Not stated
- Purity, including information on contaminants, isomers, etc.: Not stated

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Not stated
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: Not stated
- Stability in the medium, i.e. sensitivity of the test material to hydrolysis and/or photolysis: Not stated
- Solubility and stability of the test material in the solvent/vehicle and the exposure medium: Not stated
- Reactivity of the test material with the incubation material used (e.g. plastic ware): Not stated

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing (e.g. warming, grinding): Not stated
- Preliminary purification step (if any): Not stated
- Final concentration of a dissolved solid, stock liquid or gel: Not stated
- Final preparation of a solid (e.g. stock crystals ground to fine powder using a mortar and pestle): Emulsion
Species:
mouse
Strain:
not specified
Sex:
not specified
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Rationale for alternative/additional species to rat: Not stated
- Source: Not stated
- Females (if applicable) nulliparous and non-pregnant: Not stated
- Rationale for use of males Not stated
- Age at study initiation: Not stated
- Weight at study initiation: 25-30g
- Fasting period before study: Not stated
- Housing: Not stated
- Historical data: Not stated
- Diet (e.g. ad libitum): Not stated
- Water (e.g. ad libitum):Not stated
- Acclimation period:Not stated
- Microbiological status when known: Not stated
- Method of randomisation in assigning animals to test and control groups: Not stated

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Not stated
- Humidity (%): Not stated
- Air changes (per hr): Not stated
- Photoperiod (hrs dark / hrs light): Not stated
Route of administration:
oral: gavage
Vehicle:
not specified
Details on oral exposure:
VEHICLE
- Concentration in vehicle: Not stated
- Amount of vehicle (if gavage): 0.2ml
- Justification for choice of vehicle:Not stated
- Lot/batch no. (if required): Not stated
- Purity: Not stated

MAXIMUM DOSE VOLUME APPLIED: Not stated

DOSAGE PREPARATION (if unusual): Material prepared as an emulsion

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Not stated
Doses:
Not stated
No. of animals per sex per dose:
6
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: Two weeks
- Frequency of observations and weighing: Observation 40-60 minutes following administration and observations occurring within a 2 week period
- Necropsy of survivors performed: Not stated
- Clinical signs including body weight: Clinical signs were reported within the results, but a comprehensive list of all monitored not reported.
- Other examinations performed: clinical signs, body weight, organ weights, histopathology, other: Not stated
Statistics:
Litchfield and Wilcoxon methods modified by Rot
Preliminary study:
Not stated
Sex:
not specified
Dose descriptor:
LD50
Remarks:
(reported in the source paper as DL50)
Effect level:
220 mg/kg bw
Based on:
not specified
Remarks:
Limited information is available for the test material
Mortality:
It is noted that that animals died during the testing period however there was limited information available.
Clinical signs:
cyanosis, adynamia was observed along with decreased breathing and reflexes.
Body weight:
Not stated
Gross pathology:
Methemoglobin formers in the blood were noted at 39-47%. Examination of internal organs showed moderate macrovesicular hepatic steatosis/fatty liver with protein degeneration of the liver cells and hyperemia of the internal organs.
Other findings:
- Organ weights: Not stated
- Histopathology: Not stated
- Potential target organs: Nervous system
- Other observations: An increase in oxygen consumption was noted with reduced rectal temperatures

Table 2: m-ABTF toxicity when introduced to the stomach 


































Dose *



Species



Mg/kg



Mmol/kg



m-ABTF



m-ABTF



DL16



Mice



120



0.74



DL50



Mice



220



1.36



DL84



Mice



405



2.52



*Statistics was processed based on probe-analysis Litchfield and Wilcoxon methods modified by Rot (M. L. Belenky, 1959)

Interpretation of results:
study cannot be used for classification
Conclusions:
Under the limited conditions of this study outlined in the published paper the test material has an LD50 of 220 mg/kg.
Executive summary:

The published paper pre-dates the existence of nationally or internationally recognised test methods. No specific details on the test method were provided. The limited information presented indicates that 0.2ml m-Aminobenzotrifluoride (m-ABTF, a synonym for alpha,alpha,alpha-trifluoro-m-toluidine) in emulsion form was introduced to the stomach of 6 mice per dose group (100 mice in total) in a single exposure. Animal death and survival was recorded over a 2-week period. 40 - 60 minutes after the administration of m-ABTF cyanosis and adynamia was observed followed by decreased breathing and reflexes. Morphological examinations of the animals indicated the manifestation of methemoglobin formation, with the internal organs exhibiting moderate macrovesicular hepatic steatosis/fatty liver and protein degeneration of the liver cells, alongside hyperaemia of the internal organs. The LD50 was considered to be 220 mg/kg bw. Due to the limited information available on the study conducted this data is not considered reliable for use in the classification of the substance according to CLP Regulation (EC) No 1272/2008

Endpoint:
acute toxicity: oral
Type of information:
other: Published paper
Adequacy of study:
disregarded due to major methodological deficiencies
Study period:
Not stated
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Remarks:
The literature paper does not provide sufficient information on the study.
Qualifier:
no guideline available
Principles of method if other than guideline:
Not stated
GLP compliance:
no
Test type:
other: Not stated
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier) and lot/batch number of test material: Not stated
- Purity, including information on contaminants, isomers, etc.: Not stated

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Not stated
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: Not stated
- Stability in the medium, i.e. sensitivity of the test material to hydrolysis and/or photolysis: Not stated
- Solubility and stability of the test material in the solvent/vehicle and the exposure medium: Not stated
- Reactivity of the test material with the incubation material used (e.g. plastic ware): Not stated

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing (e.g. warming, grinding): Not stated
- Preliminary purification step (if any): Not stated
- Final concentration of a dissolved solid, stock liquid or gel: Not stated
- Final preparation of a solid (e.g. stock crystals ground to fine powder using a mortar and pestle): Not stated
Species:
rabbit
Strain:
not specified
Sex:
not specified
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Rationale for alternative/additional species to rat: Not stated
- Source: Not stated
- Females (if applicable) nulliparous and non-pregnant: Not stated
- Rationale for use of males Not stated
- Age at study initiation: Not stated
- Weight at study initiation: Not stated
- Fasting period before study: Not stated
- Housing: Not stated
- Historical data: Not stated
- Diet (e.g. ad libitum): Not stated
- Water (e.g. ad libitum):Not stated
- Acclimation period:Not stated
- Microbiological status when known: Not stated
- Method of randomisation in assigning animals to test and control groups: Not stated

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Not stated
- Humidity (%): Not stated
- Air changes (per hr): Not stated
- Photoperiod (hrs dark / hrs light): Not stated
Route of administration:
oral: gavage
Vehicle:
not specified
Details on oral exposure:
VEHICLE
- Concentration in vehicle: Not stated
- Amount of vehicle (if gavage): Not stated
- Justification for choice of vehicle:Not stated
- Lot/batch no. (if required): Not stated
- Purity: Not stated

MAXIMUM DOSE VOLUME APPLIED: Not stated

DOSAGE PREPARATION (if unusual): Not stated

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Not stated
Doses:
Not stated
No. of animals per sex per dose:
Not stated
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: Not stated
- Frequency of observations and weighing: Not stated
- Necropsy of survivors performed: Not stated
- Clinical signs including body weight: not stated
- Other examinations performed: clinical signs, body weight, organ weights, histopathology, other: Not stated
Statistics:
Litchfield and Wilcoxon methods modified by Rot.
Preliminary study:
Not stated
Sex:
not specified
Dose descriptor:
LD50
Remarks:
(reported in the source paper as DL50)
Effect level:
615 mg/kg bw
Based on:
not specified
Remarks:
Limited information is available for the test material
Mortality:
Not stated
Clinical signs:
Not stated
Body weight:
Not stated
Gross pathology:
Not stated
Other findings:
- Organ weights: Not stated
- Histopathology: Not stated
- Potential target organs: Not stated
- Other observations:Not stated
Interpretation of results:
study cannot be used for classification
Conclusions:
Under the limited conditions of this study outlined in the published paper the test material has an LD50 of 615 mg/kg bw.
Executive summary:

The published paper pre-dates the existence of nationally or internationally recognised test methods. No specific details on the test method were provided. No details on the test method were provided in the published paper. The LD50 was considered to be 615 mg/kg bw. Due to the limited information available on the study conducted this data is not considered reliable for use in the classification of the substance according to CLP Regulation (EC) No 1272/2008. 

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11 January 2011 to 08 February 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
This study was conducted in accordance with OECD Guideline 420 "Acute Oral Toxicity - Fixed Dose Method" (adopted 17 December 2001). This study was also performed in compliance with UK GLP standards (Schedule 1, Good Laboratory Practice Regulations 1999 (SI 1999/3106 as amended by SI 2004/0994)).
Qualifier:
according to guideline
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
fixed dose procedure
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
Female Wistar (RccHan: WIST) strain rats were supplied by Harlan Laboratories U.K. Ltd, Oxon, UK. On receipt the animals were randomly allocated to cages. The females were nulliparous and non-pregnant. After an accimatisation period of at least five days the animals were selected at random and given a number unique within the study by indelible ink-marking on the tail and a number written on a cage card. At the start of the study the animals were eight to twelve weeks of age. The bodyweight variation did not exceed +/- 20 % of the initial/mean bodyweight of any previously dosed animal(s).
The animals were housed in groups of up to four in suspended solid-floor polypropylene cages furnished with woodflakes. With the exception of an overnight fast immediately before dosing and for approximately three to four hours after dosing, free access to mains drinking water and food (2014C Teklad Global Rodent diet supplied by Harlan Laboratories UK Ltd., Oxon, UK) was allowed throughout the study. The diet, drinking water and bedding were routinely analysed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.
The temperature and relative humidity were set to achieve limits of 19 to 25 degrees celsius and 30 to 70 % respectively. Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.
The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on oral exposure:
All animals were dosed once only by gavage using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to its fasted bodyweight at the time of dosing.
Doses:
300 mg/kg was chosen as the starting dose.
No. of animals per sex per dose:
5 animals
Control animals:
no
Details on study design:
Clinical observations were made 0.5, 1, 2 and 4 hours after dosing and subsequently once daily for fourteen days. Morbidity and mortality checks were made twice daily.
Individual bodyweights were recorded on Day 0 (the day of dosing) and on Days 7 and 14.
At the end of the observation period the animals were killed by cervical dislocation. All animals were subjected to gross necropsy. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

Evaluation of data included identification of the number of animals that died during the study (or that were killed for humane reasons), and determination of the nature, severity, onset and duration of the toxic effects. If possible, the signs of evident toxicity were described. Evident toxicity referd to the toxic effects of sufficient severity that administration of the next higher dose level could result in development of severe signs of toxicity and probable mortality. Effects on bodyweights and abnormalities noted at necropsy were also identified.

Using the mortality data obtained, an estimate of the acute oral median lethal dose (LD50) of the test item was made.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 mg/kg bw
Based on:
test mat.
Mortality:
There were no deaths.
Clinical signs:
Signs of systemic toxicity noted were cyanosis, darkened eyes, pallor of the extremities, pilo-erection, hunched posture, ataxia, lethargy and pale faeces. Animals appeared normal five or eight days after dosing.
Body weight:
Individual bodyweights and bodyweight changes are given in Table 2.
Animals showed expected gains in bodyweight during the study, except for one animal which showed bodyweight loss during the first week but expected gain in bodyweight during the second week.
Gross pathology:
Individual necropsy findings are presented in Table 3.

Table 1. Individual Clinical Observations and Mortality Data

Refer to attached "background material".

Table 2. Individual Bodyweights and Bodyweight Changes

Dose Level (mg/kg) Animal Number and Sex Bodyweight (g) at day     Bodyweight Gain (g) During Week
 0 7  14  1  2
             300  1 -0 Female  169  172  189  3  17
 2 -0 Female  163  172  189  9  17
 2 -1 Female  162  166  178  4  12
 2 -2 Female  174  183  193  9  10
 2 -3 Female  165  163  187  -2  24

Table 3. Individual Necropsy Findings

 Dose Level (mg/kg)  Animal Number and Sex  Time of Death  Macroscopic Observations
             300  1 -0 Female  Killed Day 14  No abnormalities noted
 2 -0 Female  Killed Day 14  No abnormalities noted
 2 -1 Female  Killed Day 14  No abnormalities noted
 2 -2 Female  Killed Day 14  No abnormalities noted
 2 -3 Female  Killed Day 14  No abnormalities noted

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be >300 mg/kg bodyweight.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
> 300 mg/kg bw
Quality of whole database:
The experimental study (Sanders, 2011) satisfies the information requirements under REACH. The three acute toxicity via the oral route studies published under Khalepo (1968) did not satisfy the information requirements under REACH. The studies were all scored as Klimisch 4 (not assignable) therefore no endpoint conclusion could be reached.

Acute toxicity: via inhalation route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: inhalation
Type of information:
other: Published Paper
Adequacy of study:
disregarded due to major methodological deficiencies
Study period:
Not stated
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Remarks:
The literature paper does not provide sufficient information on the study
Qualifier:
no guideline available
Principles of method if other than guideline:
No guideline indicated with no information available
GLP compliance:
no
Test type:
other: Not indicated
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier) and lot/batch number of test material: Not stated
- Purity, including information on contaminants, isomers, etc.: Not stated

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Not stated
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: Not stated
- Stability in the medium, i.e. sensitivity of the test material to hydrolysis and/or photolysis: Not stated
- Solubility and stability of the test material in the solvent/vehicle and the exposure medium: Not stated
- Reactivity of the test material with the incubation material used (e.g. plastic ware): Not stated

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing (e.g. warming, grinding): Transformation to aerosol - method not stated
- Preliminary purification step (if any): Not stated
- Final concentration of a dissolved solid, stock liquid or gel: Not stated
- Final preparation of a solid: NA

FORM AS APPLIED IN THE TEST (if different from that of starting material)
- Specify the relevant form characteristics if different from those in the starting material, such as state of aggregation, shape of particles or particle size distribution: The test material is used in aerosol form, no other details are provided.
Species:
rat
Strain:
not specified
Sex:
not specified
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Rationale for alternative/additional species to rat: Not stated
- Source: Not stated
- Females (if applicable) nulliparous and non-pregnant: Not stated
- Rationale for use of males: Not stated
- Age at study initiation: Not stated
- Weight at study initiation: Not stated
- Fasting period before study: Not stated
- Housing: Not stated
- Historical data: Not stated
- Diet : Not stated
- Water: Not stated
- Acclimation period: Not stated
- Microbiological status: Not stated
- Method of randomisation in assigning animals to test and control groups: Not stated

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Not stated
- Humidity (%): Not stated
- Air changes (per hr): Not stated
- Photoperiod (hrs dark / hrs light): Not stated
Route of administration:
inhalation
Type of inhalation exposure:
not specified
Vehicle:
not specified
Remark on MMAD/GSD:
Not stated
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Not stated
- Exposure chamber volume: Not stated
- Method of holding animals in test chamber: Not stated
- Source and rate of air (airflow): Not stated
- Method of conditioning air: Not stated
- System of generating particulates/aerosols: Not stated
- Method of particle size determination: Not stated
- Treatment of exhaust air: Not stated
- Temperature, humidity, pressure in air chamber: Temperature 28˚C

TEST ATMOSPHERE
- Brief description of analytical method and equipment used: Not stated
- Samples taken from breathing zone: Not stated
- Time needed for equilibrium of exposure concentration before animal exposure: Not stated

VEHICLE
- Composition of vehicle (if applicable): Not stated
- Concentration of test material in vehicle (if applicable): Not stated
- Justification of choice of vehicle: Not stated
- Lot/batch no. (if required): Not stated
- Purity: Not stated

TEST ATMOSPHERE (if not tabulated)
- Particle size distribution: Not stated
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): Not stated

CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: Not stated
Analytical verification of test atmosphere concentrations:
not specified
Duration of exposure:
4 h
Remarks on duration:
Limited information available
Concentrations:
Not stated
No. of animals per sex per dose:
Not stated
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: Not stated
- Frequency of observations and weighing: Not stated
- Necropsy of survivors performed: Not stated
- Clinical signs including body weight: Not stated
- Other examinations performed: clinical signs, body weight, organ weights, histopathology, other: Not stated
Statistics:
Litchfield and Wilcoxon method modified by Rot
Sex:
not specified
Dose descriptor:
LC50
Remarks:
The LC50 is indicated as concentration in mg/L and correlation CL54/CL16. In addition, the ratio of possible inhalation poisoning for m-ABTF is reported by the authors as 11.9. (reported in the source paper as CL50)
Effect level:
0.44 other: mg/L and correlation CL54/CL16
Based on:
not specified
Mortality:
Not stated
Clinical signs:
other: Not stated
Body weight:
Not stated
Gross pathology:
Not Stated
Other findings:
Not stated

Table 2: m-ABTF toxicuty at a single inhalation dose 































Toxicity parameter



Animal species



Concentration in mg/l and correlation CL54/CL16



CL16



Rats



0.38



CL50



Rats



0.44



CL84



Rats



0.53



CL16



Rats



1.38



The above table indicate the LC values folllowing the inhalation of m-ABTF. The table indicates CL values which is due to the paper initially published in the Russian language where these abbreviations were not translated. The CL16 and CL84 are values at which are 1 standard deviation either side of the mean value. 


 


 

Interpretation of results:
study cannot be used for classification
Conclusions:
Under the limited conditions of this study outlined in the published paper the test material has an LC50 of 0.44 mg/L.
Executive summary:

The published paper pre-dates the existence of nationally or internationally recognised test methods. No specific details on the test method were provided. The limited information presented indicates that the rats were exposed to m-Aminobenzotrifluoride (m-ABTF, a synonym for alpha,alpha,alpha-trifluoro-m-toluidine) for 4 hours. No clincial signs were noted during the test period. The LC50 indicated as  mg/L and correlation CL54/CL16 was considered to be 0.44. In addition, the ratio of possible inhalation poisoning for m-ABTF is reported by the authors as 11.9 Due to the limited information available on the study conducted this data is not considered reliable for use in the classification of the substance according to CLP Regulation (EC) No 1272/2008.

Endpoint:
acute toxicity: inhalation
Type of information:
other: Published Paper
Adequacy of study:
disregarded due to major methodological deficiencies
Study period:
Not stated
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Remarks:
The literature paper does not provide sufficient information on the study.
Qualifier:
no guideline available
Principles of method if other than guideline:
The principles of the method were not stated.
GLP compliance:
no
Test type:
other: Not stated
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier) and lot/batch number of test material: Not stated
- Purity, including information on contaminants, isomers, etc.: Not stated

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Not stated
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions and during storage: Not stated
- Stability in the medium, i.e. sensitivity of the test material to hydrolysis and/or photolysis: Not stated
- Solubility and stability of the test material in the solvent/vehicle and the exposure medium: Not stated
- Reactivity of the test material with the incubation material used (e.g. plastic ware): Not stated

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing (e.g. warming, grinding): Transformation to aerosol, method not indicated.
- Preliminary purification step (if any): Not stated
- Final concentration of a dissolved solid, stock liquid or gel: Not stated
- Final preparation of a solid : NA

FORM AS APPLIED IN THE TEST (if different from that of starting material)
- Specify the relevant form characteristics if different from those in the starting material: Not stated
Species:
mouse
Strain:
not specified
Sex:
not specified
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Rationale for alternative/additional species to rat: Not stated
- Source: Not stated
- Females (if applicable) nulliparous and non-pregnant: Not stated
- Rationale for use of males: Not stated
- Age at study initiation: Not stated
- Weight at study initiation: Not stated
- Fasting period before study: Not stated
- Housing:Not stated
- Historical data: Not stated
- Diet (e.g. ad libitum): Not stated
- Water (e.g. ad libitum):Not stated
- Acclimation period: Not stated
- Microbiological status when known: Not stated
- Method of randomisation in assigning animals to test and control groups: Not stated

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Not stated
- Humidity (%): Not stated
- Air changes (per hr): Not stated
- Photoperiod (hrs dark / hrs light): Not stated
Route of administration:
inhalation
Type of inhalation exposure:
not specified
Vehicle:
not specified
Remark on MMAD/GSD:
MMAD and GSD not stated
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Not stated
- Exposure chamber volume: Not stated
- Method of holding animals in test chamber: Not stated
- Source and rate of air (airflow): Not stated
- Method of conditioning air: Not stated
- System of generating particulates/aerosols: Not stated
- Method of particle size determination: Not stated
- Treatment of exhaust air: Not stated
- Temperature, humidity, pressure in air chamber: 28˚C

TEST ATMOSPHERE
- Brief description of analytical method and equipment used: Not stated
- Samples taken from breathing zone: Not stated
- Time needed for equilibrium of exposure concentration before animal exposure: Not stated

VEHICLE
- Composition of vehicle (if applicable): Not stated
- Concentration of test material in vehicle (if applicable): Not stated
- Justification of choice of vehicle: Not stated
- Lot/batch no. (if required): Not stated
- Purity: Not stated

TEST ATMOSPHERE (if not tabulated)
- Particle size distribution: Not stated
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): Not stated

CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: Not stated
Analytical verification of test atmosphere concentrations:
not specified
Duration of exposure:
2 h
Concentrations:
Not stated
No. of animals per sex per dose:
Not stated
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: Not stated
- Frequency of observations and weighing: Not stated
- Necropsy of survivors performed: Not stated
- Clinical signs including body weight: Not stated
- Other examinations performed: clinical signs, body weight, organ weights, histopathology, other: Not stated
Statistics:
Litchfield and Wilcoxon method modified by Rot
Sex:
not specified
Dose descriptor:
LC50
Remarks:
The LC50 is indicated as concentration in mg/L and correlation C54/C16. (reported in the source paper as CL50).
Effect level:
0.69 other: Concentration in mg/l and correlation CL54/CL16
Based on:
not specified
Mortality:
Not stated
Clinical signs:
other: Not stated
Body weight:
Not stated
Gross pathology:
Not stated
Other findings:
Not stated

Table 2: m-ABTF toxicity at a single inhalation impact 































Toxicity parameter



Animal species



Concentration in mg/l and correlation CL54/CL16



CL16



Mice



0.54



CL50



Mice



0.69



CL84



Mice



0.88



CL84



Mice



1.63



 


 


 

Interpretation of results:
study cannot be used for classification
Conclusions:
Under the limited conditions of this study outlined in the published paper the test material has an LC50 of 0.69 mg/L.
Executive summary:

The published paper pre-dates the existence of nationally or internationally recognised test methods. No specific details on the test method were provided. The limited information indicates that the mice were exposed to m-Aminobenzotrifluoride (m-ABTF, a synonym for alpha,alpha,alpha-trifluoro-m-toluidine) for 2 hours. No clinical signs were noted during the test period. The LD50 indicated as mg/L and cprrelation CL54/C16 was considered to be 0.69. In addition, the ratio of possible inhalation poisioning for m-ABTF is reported by the authors as 11.9. . Due to the limited information available on the study conducted this data is not considered reliable for use in the classification of the substance according to CLP Regulation (EC) No 1272/2008. 

Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
The two acute toxicity via inhalation route studies published under Khalepo (1968) did not satisfy the information requirements under REACH. The studies were all scored as Klimisch 4 (not assignable) therefore no endpoint conclusion could be reached.

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
other: Published Paper
Adequacy of study:
disregarded due to major methodological deficiencies
Study period:
Not stated
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Remarks:
The literature paper does not provide sufficient information on the study.
Qualifier:
no guideline available
Principles of method if other than guideline:
-Principle of the test: The acute dermal toxicity potential of the m-ABTF was evaluated on white mice
- Short description of test conditions: 2/3 of the animal’s tail was submerged into the substance for various periods of exposure (1-8 hours).
- Parameters analysed / observed: Skin restorative effect was assessed based on the mortality outcome, calculating the time of death of 50% of the studied animals.
GLP compliance:
no
Test type:
other: Not stated
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier) and lot/batch number of test material: Not stated
- Purity, including information on contaminants, isomers, etc.: Not stated

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Not stated
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: Not stated
- Stability in the medium, i.e. sensitivity of the test material to hydrolysis and/or photolysis: Not stated
- Solubility and stability of the test material in the solvent/vehicle and the exposure medium: Not stated
- Reactivity of the test material with the incubation material used (e.g. plastic ware): Not stated

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing (e.g. warming, grinding): NA
- Preliminary purification step (if any): Not stated
- Final concentration of a dissolved solid, stock liquid or gel: undiluted
- Final preparation of a solid (e.g. stock crystals ground to fine powder using a mortar and pestle): NA
Species:
mouse
Strain:
not specified
Sex:
not specified
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Rationale for alternative/additional species to rat (if applicable): Not stated
- Source: Not stated
- Females (if applicable) nulliparous and non-pregnant:Not stated
- Rationale for use of males (if applicable): Not stated
- Age at study initiation: Not stated
- Weight at study initiation: Not stated
- Fasting period before study: Not stated
- Housing:Not stated
- Historical data:Not stated
- Diet (e.g. ad libitum): Not stated
- Water (e.g. ad libitum):Not stated
- Acclimation period:Not stated
- Microbiological status when known: Not stated
- Method of randomisation in assigning animals to test and control groups: Not stated

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Not stated
- Humidity (%): Not stated
- Air changes (per hr): Not stated
- Photoperiod (hrs dark / hrs light): Not stated
Type of coverage:
other: 2/3 of the tail was submerged in the substance
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: 2/3 of the tail
- % coverage: Not stated
- Type of wrap if used: NA

REMOVAL OF TEST SUBSTANCE
- Washing (if done): NA
- Time after start of exposure: Not stated

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Not stated
- Concentration (if solution): Pure - concentration not specified
- Constant volume or concentration used: Not stated
- For solids, paste formed: yNA

VEHICLE
- Amount(s) applied (volume or weight with unit): NA
- Concentration (if solution): NA
- Lot/batch no. (if required): NA
- Purity: NA
Duration of exposure:
Between 1-8 hours
Doses:
Not stated
No. of animals per sex per dose:
Not stated
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: Not stated
- Frequency of observations and weighing: Not stated
- Necropsy of survivors performed: Not stated
- Clinical signs including body weight: Not stated
- Other examinations performed: clinical signs, body weight, organ weights, histopathology, other: The skin restorative effect was examined.
Statistics:
Not stated
Key result
Sex:
not specified
Dose descriptor:
other: TL50
Effect level:
6 other: hours
Based on:
test mat.
Remarks on result:
other: TL = time of death of 50% of animals
Mortality:
Mortality was noted with limited information available to comment further
Clinical signs:
Not stated
Body weight:

Not stated
Gross pathology:
Not stated
Other findings:
In surviving animals, the tail was subjected to dry necrosis with it falling off on day 3-4.
Interpretation of results:
study cannot be used for classification
Conclusions:
Under the limited conditions of this study outlined in the published paper the test material has an TL50 of 6 hours.
Executive summary:

The published paper pre-dates the existence of nationally or internationally recognised test methods. No specific details on the test method were provided. The limited information presented indicates that the tails of mice were submerged into the test substance m-Aminobenzotrifluoride (m-ABTF, a synonym for alpha,alpha,alpha-trifluoro-m-toluidine) in a single exposure for a period of 1 - 8 hours. The effect on skin of the tail was noted with animal dealth and survival recoreded. The TL50 was determined to be 6 hours with tails of surviving animals showing dry necrosis with sections of exposed tail falling off on day 3-4. Due to the limited information available on the study conducted this data is not considered reliable for use in the classification of the substance according to CLP Regulation (EC) No 1272/2008.

Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
The acute dermal via dermal route study published under Khalepo (1968) did not satisfy the information requirements under REACH. The studies were all scored as Klimisch 4 (not assignable) therefore no endpoint conclusion could be reached.

Additional information

Assessment of the test substance for acute toxicity was published within a paper which reported toxicity of fluorine derivatives of toluene (multiple related substances, of which alpha,alpha,alpha-trifluoro-m-toluidine was one) following single exposures in a number of animal models (Khalepo, 1968). The acute oral toxicity testing on rats, mice and rabbits within Khalepo (1968) did not provide adequate detail on the studies, therefore these studies were considered to be unreliable and assigned a Klimisch Score of 4. As such, it was not possible to determine a classification. Similar inadequacies were noted in both the acute inhalation and the acute dermal study performed on rats, mice and rabbits respectively due the limited and unreliable data available. As a result, these were also assigned a Klimisch score of 4. In summary, the acute toxicity results reported by Khalepo (1968) were not adequate for the classification of the test substance.


As the results reported by Khalepo (1968) did not provide adequate information to classify the substance, it was necessary to complete an in vivo oral study (Sanders 2011). The study was performed to OECD TG 420 (Acute oral toxicity - fixed dose method) under GLP conditions to assess the acute oral toxicity potential of the test material when administered by oral gavage to Female Wistar rats. 300 mg/kg bw of the test material in a vehicle of arches oil was administered. Assessment of acute toxicity was made at approx. 0.5, 1, 2 and 4 hours following administration, and then once daily for 14 days. Morbidity and mortality checks were undertaken twice daily.


The administration of test material by oral gavage did not produce any deaths during the test period. Signs of systemic toxicity were noted, manifesting as cyanosis, darkened eyes, pallor of the extremities, pilo- erection, hunched posture, ataxia, lethargy and pale faeces. The animals were noted as 5 or 8 days after dosing. No abnormalities were noted at necropsy of all animals.


The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was therefore estimated to be >300 mg/kg bodyweight. As the oral LD50 was >300 but ≤2000 mg/kg bodyweight, the test substance is determined to be an Acute oral tox. category 4 according to CLP Regulation (EC) No 1272/2008. 


 

Justification for classification or non-classification




Results from the key study OECD TG 420 (Sanders, 2011) were sufficient for Category 4 Acute Oral tox classification according to CLP Regulation (EC) No 1272/2008.