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Administrative data

Description of key information

 The LD50 of the test substance was higher than or equal to 2000 mg/kg by dermal or oral exposure. Fully described under "Discussion".

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14 days
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The report satisfies the GLP regulations and specific test guidelines in general, except that the concentration/purity of the test substance could not be directly verified from the Certificate of Analysis.
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Version / remarks:
EU B.1 ofAnnex V was also followed
Deviations:
yes
Remarks:
1. Test substance characterization and stability data, while available, were not developed in accordance with GLP. Report stated that stability was not expected to impact the study results.
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
other: CD strain (remote Sprague-Dawley origin) from Charles River (UK) Limited, Kent, England.
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Limited, Margate, Kent, England
- Age at study initiation: Five weeks old
- Weight at study initiation: males, 122 - 133 g; females, 114 - 138 g on the day prior to initiation.
- Fasting period before study: 17 hours before dosing
- Housing: Five animals of the same sex per stainless steel grid cage
- Diet (e.g. ad libitum): commercially-available, pelleted rodent diet was available ad libitum.
- Water (e.g. ad libitum): Free access to tap water
- Acclimation period: Atleast five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-21°C
- Humidity (%): 38 -57%
- Air changes (per hr): 15
- Photoperiod: 12 hrs dark / 12 hrs light

IN-LIFE DATES: From 14 October, 1992 to 5 Nov, 1992
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
Test formulation was prepared at the appropriate concentration in purified water shortly before administration. Oral gavage was carried out with a flexible rubber catheter allowing instillation of the dose into the lumen of the stomach into overnight fasted rats. Food was made available three hours after dosing. On the basis of the preliminary results, the main study was done.
Doses:
One dose, 2000 mg/kg [the maximum practical dose]
No. of animals per sex per dose:
Preliminary Study: one male and one female rat given a single oral dose of 800 mg/kg at a constant dose-volume of 20mL/kg.

Main study: five rats of each sex per dose at a constant dose-volume of 20mL/kg.
Control animals:
no
Details on study design:
Animals were observed daily (five inspections on Day 1; twice per day from Day 2 onwards). Body weights were recorded on the day before dosing and on Days 1,8 and 15. All animals were necropsied on Day 15.
Statistics:
None
Preliminary study:
No death noticed.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
One male rat died three hours after dosing on Day 1 in the main study.
Clinical signs:
Ante mortem signs of the animal that died comprised lethargy, staggering gait, prone position, bradypnoea, hyperpnoea, piloerection, and salivation.

Signs of reaction to treatment in the surviving animals comprised of underactivity, staggering gait, piloerection, salivation, and hunched posture. The surviving animals were overly normal on the third day after treatment.
Body weight:
The surviving animals achieved expected bodyweight gain.
Gross pathology:
Necropsy of the decedent animal (male) revealed pale muzzule staining. Necropsy of the surviving animals, on Day 15, revealed no significant macroscopic lesion.
Interpretation of results:
sligthly toxic
Remarks:
Migrated information Criteria used for interpretation of results: OECD GHS
Conclusions:
Under the conditions of this study, the oral median lethal dosage (LD50) of the test material was greater than 2000 mg/kg.
Executive summary:

Acute oral toxicity of Trigonox 44B was investigated in a fourteen-day study in five male and female CD rats after a single oral gavage dose of 2000 mg/kg. One male rat showed treatment-related clinical signs and died three hours after dosing, but the rest of the animals survived till the end of the study. Surviving animals were overtly normal on the third day after treatment, but none of the surviving animals showed an effect of Trigonox 44B on body weight gain or macroscopic lesions at necropsy. Under the conditions of this study, the oral median lethal dosage (LD50) of the test material was greater than 2000 mg/kg.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw
Quality of whole database:
K2. An apparently well done study. The LD50 was determined to be greater than 2000 mg/kg.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Although the study report did not cite the test guideline followed (OECD test guidelines were not available then), the brief description of the experiment appear reasonable. The test item's identity was not clearly identified by CAS number in the report, and had to be inferred. Only one concentration of the test item was tested and for an hour. These approaches differ from the currently acceptable test guidelines. Therefore the experimental value cannot be used for DNEL derivation.
Qualifier:
no guideline followed
Principles of method if other than guideline:
Rats were exposed to a single continuous aerosol mist of the test compound for an hour in a dynamic inhalation chamber. Exposure was made by metering the test material into a nebulizer. Animals were observed for 9 days followed by necropsy.
GLP compliance:
no
Remarks:
This is an old study (1965), OECD test guidelines were not available at that time.
Test type:
fixed concentration procedure
Limit test:
yes
Species:
rat
Strain:
other: Albino Charles River rats were used.
Sex:
male
Details on test animals and environmental conditions:
Ten male albino Charles River rats, weighing from 264 to 296 grams, were used in this study. The animals were individually housed in wire mesh cages elevated above the droppings and maintained in airconditioned quarters throughout the pre-exposure and 2-week postexposure period. Except for the period of exposure, food and water were available to all animals ad libitum.
Route of administration:
other: Aerosol mist
Type of inhalation exposure:
whole body
Remarks:
The aerosol formed was carried directly into the exposure chamber containing the test material.
Vehicle:
not specified
Details on inhalation exposure:
The test animals were placed in an 18 liter, dynamic inhalation chamber and exposed, for a single continuous one-hour period, to the aerosol mist of the test material. Exposure was made by metering the test materials into a nebulizer (DeVilbiss No. 40) through which a known airflow was produced. The aerosol thus formed was carried directly into the exposure chamber containing the test animals.

The apparatus used was fitted with a rotameter and a Dow Dual Syringe Feeder, permitting variable airflow and an alteration of the flow of the test material into the nebulizer. Calculations of the theoretical concentrations were based on the values obtained from these instruments.

The atmospheric concentration used during this exposure (13.1 mg./liter) was the maximum possible using the apparatus described.
Analytical verification of test atmosphere concentrations:
no
Concentrations:
The test compound, SN4-l38-38, was administered as an aerosol mist at a calculated atmospheric concentration of 13.1 mg./liter of air. The atmospheric concentration used during this exposure (13.1 mg./liter) was the maximum possible using the apparatus described.
No. of animals per sex per dose:
Ten male rats were used. Only one dose was tested.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 9 days. During the l-hour exposure to an atmosphere containing an aerosol mist of the test compound, all animals were observed continuously for changes in behavior and appearance.. Following the exposure, the animals were examined closely for pharmacodynamic and/or toxic signs.

- Frequency of observations and weighing: Prior to the initiation of the exposure, and at 7 and 9 days thereafter, individual body weight measurements were obtained.

- Necropsy of survivors performed: yes. After 9 days of observations, all rats were sacrificed by means of an intraperitoneal injection of 5 per cent sodium pentobarbital and necropsied.

- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:
Sex:
male
Dose descriptor:
discriminating conc.
Effect level:
> 13.1 mg/L air
Based on:
test mat.
Exp. duration:
60 min
Mortality:
No mortalities occurred.
Clinical signs:
other: Behavior, and Appearance: Gross observation of the test animals after 10 minutes of the exposure period had lapsed revealed 3-of-10 with moderate ptyalism and a general increased grooming activity throughout the group. After 30 minutes of exposure, all ra
Body weight:
Individual body weights taken initially and at 7 and 9 days after treatment did not reflect unusual alterations and compared favorably to control rats of the same age and strain maintained in the testing laboratory from time-to-time.
Gross pathology:
Necropsy Findings: No lesions were observed at necropsy which could be attributed to the inhalation exposure with the test compound.
Interpretation of results:
study cannot be used for classification
Remarks:
Criteria used for interpretation of results: expert judgment
Conclusions:
Exposure of rats for one continuous hour to an atmospheric concentration of a mist of the test agent (13.1 mg./liter of air) failed to cause adverse pharmacotoxic signs, mortality, or gross evidence of damage at necropsy 9 days after exposure.
Executive summary:

Ten male albino Charles River rats, weighing from 264 to 296 grams, were used in this study. The test compound, SN4-l38-38, was administered as an aerosol mist at a calculated atmospheric concentration of 13.1 mg./liter of air. During the l-hour exposure all animals were observed continuously for changes in behavior and appearance. Following the exposure, the animals were examined closely for pharmacodynamic and/or toxic signs. After 9 days of observations, and body weight measurements on days 7 and 9, animals were necropsied.

Exposure of rats for one continuous hour to an atmospheric concentration of a mist of the test agent (13.1 mg./liter of air) failed to cause adverse pharmacotoxic signs, mortality, or gross evidence of damage at necropsy 9 days after exposure. Based on the results of this study, it was concluded that the results indicate that the test compound does not present a hazard to rats when a saturated atmosphere is inhaled over a continuous period of one hour. The report also concluded that dependent upon the use of this agent, there does not appear to be a hazard to man by the route of inhalation provided ordinary care is exercised in the handling of this material.

Only one concentration of the test item was tested and for an hour. These approaches differ from the currently acceptable test guidelines. Therefore the experimental value cannot be used for DNEL derivation.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating conc.
Value:
13.1 mg/m³
Quality of whole database:
K2. An apparently well done study. There was no toxicity observed at the dose of 13.1 mg/liter..

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: An apparently well done GLP study performed according to OECD test guideline 402.
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
not specified
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
Animals
Species, strain: rat, Sprague-Dawley ICO: OFA-SD (lOPS Caw).
Reason for this choice: rodent species cOIl11ilonly requested by the international regulations for this type of study.
Breeder: Iffa Credo, 69210 L' Arbresle, France.
Number and sex: 1 group of 10 animals (5 males and 5 females).
Age/weight: on the day of treatment, the animals were approximately 8 weeks old, and had a mean body weight of 256 ± 12 g for the males and 219 ± 4 g for the females.
Acclimatization: at least 5 days before the beginning of the study.
Identification of the animals: the animals were identified individually by earmarks or earnotches.

Environmental conditions
During the acclimatization period and during the main test, the conditions in the animal room were as follows:
temperature: 21 ± 2°C
relative humidity: 30 to 70%
light/dark cycle: 12 hl12 h
ventilation: about 12 cycleslhour of filtered, non-recycled air.
The temperature and relative humidity were recorded continuously and records retained.
The housing conditions (temperature, relative humidity, light/dark cycle and ventilation) were checked monthly. The animals were housed in polycarbonate cages covered with a stainless steel lid.
During the acclimatization period each cage (48 x 27 x 20 cm) contained 4 to 7 animals of the same sex. During the treatment period each cage (35.5 x 23.5 x 19.3 cm) contained 1 rat. Each cage
contained graded, dust-free sawdust (SICSA, 94142 Alfortville, France). Bacteriological analysis of the sawdust and detection of possible contaminants (pesticides, heavy metals) are performed periodically.

Food and water
All the animals had free access to A04 C pelleted diet (U.A.R., 91360 Villemoisson-sur-Orge, France). Each batch of food was analysed (composition and contaminants) by the supplier. The diet formula is presented in appendix 2.

Drinking water filtered by a F.G. Millipore membrane (0.22 micron) was contained in bottles. Bacteriological and chemical analysis of the water and detection of possible contaminants (pesticides, heavy metals and nitrosamines) are performed periodically. Results are archived at C.LT. There were no contaminants in the diet, water or sawdust at levels likely to have influenced the
outcome of the study.

Type of coverage:
occlusive
Details on dermal exposure:
TREATMENT

Preparation of the animals
On the day before treatment, the dorsal area (6 cm x 8 cm) of each animal was clipped of hair using electric clippers. Only animals with healthy intact skin were used for the study.

Mode of treatment
As the test substance was anticipated to be non-toxic at 2000 mg/kg, a limit test was performed by application of 2000 mg/kg of the test substance to a group of 10 animals (5 males and 5 females).

Taking into consideration that the specific gravity (SO) of the test substance was 1.025, the test substance in its original form and at a dose of 2000 mg/kg, was applied directly to an area of the skin representing approximately 10% (5 x 6 cm for the females and 5 x 7 cm for the males) of the body surface of the animals. This was calculated according to Meeh's formula (1). A hydrophilic gauze pad (Semes France, 54183 Heillecourt, France) was then applied to the skin. The test substance and the gauze pad were held in contact with the skin for 24 hours by means of an adhesive hypoallergenic aerated semi-occlusive dressing (Laboratoires de Pansements et d'Hygiene, 21300 Chenove, France) and a restraining bandage (Laboratoires 3M Sante, 92245 Malakoff, France). This dressing prevented ingestion of the test substance by the animals. No residual test substance was observed at removal of the dressing.
Duration of exposure:
Date of treatment
The single administration was performed on 1.9.94 in the morning (day 1) and was followed by
a 14-day observation period until 15.9.94 (day 15).
Doses:
As the test substance was anticipated to be non-toxic at 2000 mg/kg, a limit test was performed by application of 2000 mg/kg of the test substance to a group of 10 animals (5 males and 5 females). The dose applied to each animal was adjusted according to body weight determined on the day
of treatment.
No. of animals per sex per dose:
5
Control animals:
yes
Details on study design:
CLINICAL EXAMINATIONS

Clinical signs
The animals were observed frequently during the hours following application of the test substance for detection of possible treatment-related clinical signs. Observation of the animals was made at least once a day for a period of 14 days, to determine whether any of the clinical signs were reversible or not.

Mortality
The animals were checked frequently during the hours following application of the test substance for mortality or signs of morbidity, then at least twice a day thereafter.

(1) Meeh's formula (AFNOR-T03-23 Standard, August 1980, France)
S =k\j"fT
S = Total surface of the animal in dm2
p = Weight of the animal in grams
k =0.09 for the rat


Body weight
The animals were weighed individually just before administration of the test substance then on days 8 and 15. The body weight gain of the treated animals was compared to a reference curve of C.LT. control animals with the same initial weight.

PATHOLOGY

Necropsy
On day 15, the animals were killed by CO2 inhalation in excess and a macroscopic examination was performed.

Macroscopic examination
After opening the thoracic and abdominal cavities, a macroscopic examination of the main organs (digestive tract, heart, kidneys, liver, lungs, pancreas, spleen and any other organs with obvious abnormalities) was performed.

Microscopic examination
No microscopic examination was performed.
Statistics:

DATA EVALUATION

Evaluation of the innoxiousness or toxicity of the test substance following a single dermal application in rats should include the relationship, if any, between the animals' exposure to the test substance and the incidence and severity of all the abnormalities including behavioural and clinical abnormalities, macroscopic lesions, body weight changes, mortality and any other toxic effects.

Classification of the test substance is based on the following criteria: Commission Directive 93/21/E.E.C.


LDso
dermal route Labelling Indication Symbol
(mg/kg) sentence of danger


<50 R27 Very toxic in contact with skin Very toxic T+
50 < and < 400 R24 Toxic in contact with skin Toxic T
400 < and < 2000 R21 Harmful in contact with skin Harmful Xn
>2000 None None None

Preliminary study:
Results
There was no preliminary study.

Results of the main study:
The general behaviour and body weight gain of the animals were not affected by treatment with the test substance. No cutaneous reactions were observed. No deaths occurred at 2000 mg/kg. A macroscopic examination revealed no abnormalities in the animals killed at the end of the study.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
>= 2 000 mg/kg bw
Based on:
test mat.
Mortality:

Mortality:
No deaths occurred during the observation period.
Clinical signs:
CLINICAL EXAMINATIONS
Clinical signs and cutaneous reactions (tables 1 and 2): No clinical signs and cutaneous reactions were observed during the study.
Body weight:
Body weight (treated animals and control animals)

Between days 1 and 8, a decrease in body weight was noted for one female (No. 01). This finding was probably due to the stress caused by the experimental conditions and was not attributed to treatment with the test substance. The body weight of this animal showed an improvement during the remainder of the observation period. The body weight gain of the remaining animals was normal.
Gross pathology:
PATHOLOGY

Macroscopic examination of the main organs of the animals killed at the end of the study revealed no apparent abnormalities.
Conclusions:
CONCLUSION

Under the experimental conditions, the LDo of the test substance, PEROXIMON K3, when administered by dermal route in rats was higher than or equal to 2000 mglkg. No signs of toxicity were observed at this dose.
Executive summary:

SUMMARY

At the request of Elf Atochem, Milano, Italia, the acute TOXICITY of the test substance, PEROXIMON K3, by dermal route was evaluated in rats according to O.E.C.D. (No. 402, 24th February 1987) and E.C. (92/69/E.E.C.). The study was conducted in compliance with the

principles of Good Laboratory Practice.

 

Methods

The test substance was administered by dermal route to a group of 10 Sprague-Dawley rats (5 males and 5 females). The test substance in its original form was applied directly to the skin of the animals at a dose of 2000 mg/kg, taking into consideration that the specific gravity (SG) of the test substance was 1.025. After 24 hours under a semi-occlusive dressing, no residual test substance was observed on removal of the dressing. The animals were checked for clinical signs, mortality and body weight gain for a period of 14 days following the single application of the test substance. A necropsy was performed on each animal killed at the end of the study.

 

Results

The general behaviour and body weight gain of the animals were not affected by treatment with the test substance. No cutaneous reactions were observed. No deaths occurred at 2000 mg/kg. A macroscopic examination revealed no abnormalities in the animals killed at the end of the study.

 

Conclusion

Under the experimental conditions, the LD50 of the test substance, PEROXIMON K3, when administered by dermal route in rats was higher than or equal to 2000 mg/kg. No signs of toxicity were observed at this dose.

 

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw
Quality of whole database:
K1. An apparently well done study. The LD50 was determined to be greater than 2000 mg/kg.

Additional information

In this study, the toxicity of the test substance, PEROXIMON K3 (37187 -22 -7), was tested by dermal route in Sprague-Dawley rats (5 males and 5 females) at a dose of 2000 mg/kg, according to OECD No. 402 (Jouffrey, 1994). The general behaviour and body weight gain of the animals were not affected by treatment with the test substance. No cutaneous reactions were observed. No deaths occurred at 2000 mg/kg. A macroscopic examination revealed no abnormalities in the animals killed at the end of the study. The LD50 of the test substance was higher than or equal to 2000mg/kg.

Acute oral toxicity of the test substance was investigated in a 14-day study in five male and female CD rats after a single oral gavage dose of 2000 mg/kg (Rees, 1993). Under the conditions of this study, the oral median lethal dosage (LD50) of the test material was greater than 2000 mg/kg. These results are in agreement with the LD50 determined by the dermal toxicity study.

Inhalation: Exposure of rats for one continuous hour to an atmospheric concentration of a mist of the test agent (13.1 mg/liter of air) failed to cause adverse pharmacotoxicological effects (Wazeter, 1965). However, the study has some experimental deficiencies.


Justification for selection of acute toxicity – oral endpoint
A well conducted and well described recent GLP study, according to OECD guideline

Justification for selection of acute toxicity – inhalation endpoint
A reasonably well conducted study, done prior to OECD test guideline availability

Justification for selection of acute toxicity – dermal endpoint
A well conducted and well described recent GLP study, according to OECD guideline

Justification for classification or non-classification

The acute toxicity study data resulted in non-classification of the test material. The acute toxicity data are conclusive but not sufficient for classification.