A mixture of: esters of C14-C15 branched alcohols with 3,5-di-t-butyl-4-hydroxyphenyl propionic acid; C15 branched and linear alkyl 3,5-bis(1,1-dimethylethyl)-4-hydroxybenzenepropanoate; C13 branched and linear alkyl 3,5-bis(1,1-dimethylethyl)-4-hydroxybenzenepropanoate

Endpoint summary

Description of key information

Acute Oral Toxicity
No signs of toxicity were seen in any animals dosed with 5000 mg/Kg of the test article. Therefore the LD50 of the test article is concluded to be >5000 mg/Kg bw.

Acute Dermal Toxicity
The test article ANOX - BF, when administered by dermal route to rat, under the conditions adopted in this experiment, did not cause mortality nor toxic effects at the limit dose of 2000 mg/Kg.
The LD50 by dermal route is higher than 2000 mg/Kg

Acute Inhalation Toxicity

ANOX BF did not produce any evidence of toxicity in Sprague-Dawley rats following exposure for 4 hours to an atmosphere containing 7.53 mg. liter -1 (measured gravimetrically).

 

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:

January 29 1991 to February 12 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: 84/449/EWG, B.1

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

Not specified

Species:

rat

Strain:

Sprague-Dawley

Remarks:

Cr1:CD (SD) BR rats

Sex:

male/female

Details on test animals or test system and environmental conditions:

Body weight (on receipt): Males: 225-250 g
Females: 200-225 g
Age (on receipt): About 7-9 weeks
Supplier: Charles River Italia S.p.A. Via Indipendenza, 11, 22050 - CALCO (Como),
Shipping slip number: 00621 dated Jan. 21, 1991
Acclimation: at least 5 days before the start of the test. Animals were observed daily to ascertain their fitness for the study.
Housing: 5 animals/sex/cage in air-conditioned rooms.
- Temperature: 22 ° C ± 2
- Relative humidity: 55% ± 10
- Air changes: about 20/hour filtered on HEPA 99.97%.
- Light: 12 hour cycle (7 a.m. - 7 p.m.)
- Cage size: grill cages cm 40.5x38.5x18h with stainless steel feeder. The waste that
dropped through the grill bottom on removable paper 􀇣as periodically disposed of.
Animal identification: By nicks or tipping's of the outer ear. Cage card gave experiment number,
dosage group, sex and date of administration.
Diet: GLP 4RF21 pelleted diet produced by Charles River Italia' s feed licensee Yucedola
Sl., Settimo Yilanese. The declared contents, on the label, on dry m a t t e r basis (moisture 12%), were:
crude protein 13.50%
crude fat 3.00%
crude fiber 6.00%
ash 7.00%
The diet was supplemented by the Producer with vitamins and trace elements. The Producer supplies a certificate of anal:-sis for nutrients and contaminants, the levels of which are within the limits proposed by EPA-TSC-4 (44FR:4053-44093, July 26, 1979). RBM has the animal feed re-analyzed at Least twice a year for bacterial contamination. The diet was available "ad libitum" to the animals.
Water: from the municipal water main system. water is filtered and distributed "act· libitum" to the animals by an automatic valve system.
Periodically drinking water is analyzed for microbiologic count. heavy metals, other contaminants (e.g. solvents, pesticides) and other chemical and physical characteristics. The acceptable limits of quality of the drinking water were those defined in EEC Directive 80/778.

Contaminants that might interfere with the objectives of the study 􀆉ere not expected to be present in diet or drinking water.

Route of administration:

oral: gavage

Vehicle:

methylcellulose

Details on oral exposure:

Preparation and administration of test article
Just before the treatment of the animals a weighed amount of test article , was suspended with the suitable volume of 0.5% methylcellulose 400 cps water solution. In order to maintain a good homogeneity the suspension was kept stirred by magnetic
stirring until the end of the treatment.
The suspension was administered at the constant volume of 20 ml/kg. The volumes to be administered were measured with appropriately gauged plastic syringes.
The administration was done by gavage to rats which had been fasted about 16 hours.
Analysis:
Stability - Not requested by the Sponsor
Concentration check - Not requested by the Sponsor

Doses:

Single dose of 5000 mg/Kg body weight.

No. of animals per sex per dose:

5 males + 5 females/group

Control animals:

not specified

Details on study design:

Administration route: oral (by gavage)
Reason for selection of administration route : Possible accidental ingestion by humans.
Administration frequency: Single.
Experimental design: The test article was administered as follows:
Dose: (mg/Kg): 5000
Administration volume (ml/Kg): 20
Treatment date: January 29 1991

Observation period: 14 days after the administration.
Observation of clinical signs and mortality: At 30 minutes, 2, 4 and 6 hours on the first day after administration and then twice a day up to termination of the observation period.

Body weight: Once pre-trial, on days 1, 3, 8 and 14. On day 1 the animals were weighed after a 16 hour fasting period. The volume of administration was based on day 1 body weight. Feed was returned to rats three hours after the test article administration.

Gross pathology: On all the animals that died during the study and all the animals (fasted overnight) killed by excision of the femoral arteries, after i.p. anaesthesia with 5% sodium pentobarbital, at the end of the observation period.

Histology: Portions of abnormal entities found in any of the necropsied animals were collected. The tissue samples were fixed and preserved in 10 % buffered formalin.
Histological examinations were not performed as no macroscopic findings emerged from necropsy.

Statistics:

The LD50 calculations were not possible.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed in animals treated at 5000 mg/Kg.

Clinical signs:

other: Signs of toxicity related to dose levels: Piloerection was observed only two hours after test article administration in two male and three female rats.

Gross pathology:

Effects on organs:
At the gross pathology examination carried out at the end of the observation period no changes were noted in all necropsied animals.

Interpretation of results:

GHS criteria not met

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

No signs of toxicity were seen in any animals dosed with 5000 mg/Kg of the test article. Therefore the LD50 of the test article is concluded to be >5000 mg/Kg bw.

Executive summary:

A acute oral toxicity study was carried out in rats treated with test article ANOX – BF,  according to test guideline 84/449/EWG, B.1, in compliance with GLP.

The test article was suspended in 0,5% methylcellulose 400 cps water

Solution and administered to rats at the volume of 20 ml/kg.

All rats were treated after a 16 hrs fasting period. The animals were weighed on days 1, 3, 8 and 14; clinically observed for 14 days following the treatment and killed at the end of the study by excision of the femoral arteries after having been completely anesthetized with an i.p. injection of 5% sodium pentobarbital . A thorough autoptic  examination was performed in animals killed at the end of the observation period .

No animals died during the observation period.

Two male and three female treated rats only showed transient piloerection 2 hours after treatment.

The body weight gain appeared unaffected by treatment .

At the gross pathology examination performed on rats killed at the end of the observation period no changes were noted.

In conclusion ANOX - BF, when administered by oral rout to rat, under' the conditions adopted in this study, did not cause overt signs of toxicity- and showed an LD50 higher than 5000 mg/kg.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 mg/kg bw

Quality of whole database:

K1

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

Study period:

May 17 1991 to June 18 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

other: Acute inhalation toxicity

Limit test:

yes

Specific details on test material used for the study:

Not specified

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Animals
5 male and 5 female Sprague-Dawley rats were received from Charles River (UK) Limited, Margate, Kent, England on 9 May 1991. A t the time of arrival the batch o f animals were within the weight range 121-150 g. A further 5 male and 5 female rats were assigned t o the study in order to dose the additional group (Group 2). These additional animals were received on 9 May 1991, in the weight range 124-150 g.

Housing
The rats were housed in a semi-barrier maintained animal room with a target room temperature of 20°C and a relative humidity of -ca 55% +/- 10%. Temperature and humidity extremes recorded were 19 - 22°C and 34 - 45% respectively. Although these environmental extremes deviated from the protocol-stated target ranges, they were not considered to have affected the integrity of the study. A 12 h light-dark cycle was controlled by a time switch; light hours being 0700-1900 h. The animals were housed in Room N21 of the Block N Rodent Inhalation Toxicology Complex at the Elphinstone Research Centre.

Animal maintenance
Rats were housed 5 per cage according to sex in suspended stainless steel cages. All cages were suspended over trays containing absorbent paper. Each cage was supplied with a polypropylene water bottle (capacity 500 ml). with a rubber washer and melamine cap. Cages and cage tray papers were changed as necessary during the study period whilst water bottles were cleaned or changed as required. Each afternoon, when other work in the room was finished, floors were swept then washed with a disinfectant solution . Approximately once each week walls, benches and racking within the animal room were washed with a disinfectant solution. The disinfectant used was 1% Tego from Th Gofdschmidt and Company Limited, Middlesex, England.

Diet
The food and water used on the study were considered to contain no contaminants insufficient quantity to have had any influence on the outcome o f the study.

Food
Rat and Mouse (Modified) No. 1 Diet SQC Expanded, supplied by Special Diets Services Limited, Stepfield, Witham, Essex was available.

Water
The animals had access to domestic mains quality drinking water.

Food and water were available adlibitum except during the 4 h exposure period.

Animal identification
Each animal was given a unique earmark which identified it within the study and corresponded to that animal's study number.
Each animal was ascribed a cage card which identified that animal by project number, animal number, sex, treatment group and cage number.

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

snout only

Vehicle:

air

Geometric standard deviation (GSD):

ca. 0.49 - ca. 3.3

Details on inhalation exposure:

The test atmospheres were generated by metering the test material via a peristaltic pump to a Schlick atomiser which was located at the top of the exposure chamber. The concentration within the exposure chamber was controlled by adjusting the rate o f feed of the test material and the air flow rate to the atomiser. Chamber air flow rates were monitored continuously and the values recorded at 30 min intervals.
Filtered, oil-free compressed air , for the production of the test atmosphere, was supplied by oi 1-free Hydrovane compressors. The aluminum exposure chamber (ADG Instruments Limited, Hitchin, Herts, England) was cylindrical in cross section and had a volume of approximately 41.5 liters . An extract duct from the chamber was connected by way of a high efficiency filter to a metered vacuum system. The exposure chamber was located inside an extract chamber for the protection of the operators and the environment.
The exposure system was truly dynamic, incorporating a single pass of the freshly generated material. The aerosol particles dispersed throughout the chamber and exited from the base through the trap to a filtered vacuum line. The inhalation exposures were conducted in a room (N19) adjacent to the animal holding room.
Prior to exposure, each animal was removed from it's cage and examined for general health status. The ear number was checked, the animal weighed and then loaded into a tapered, polycarbonate restraint tube which fitted onto the exposure chamber and sealed by means of a push fit through a rubber '0' ring. All the animals were exposed on a single tier to eliminate any exposure variation. Only the animals' snouts were exposed to the test atmosphere. The animals were not allowed access t o food or water during the 4 h exposure period.
During the exposure the animals were observed at regular intervals for signs of any adverse reactions to treatment. On completion of the 4 h exposure period, the animals were removed from the chamber, unloaded from the restraint tubes, returned t o their cages in the animal holding room and observed for clinical signs.
The Group 1 animals (5 males and 5 females) were exposed to 4.07 mg liter -1 while trying to achieve the target limit concentration of 5 mg liter -1 via the inhalation route by snout only exposure for a single continuous 4 h period. The exposure was conducted on 17 May
1991.
The Group 2 animals were dosed to ensure the target limit was achieved and were exposed to 7.53 mg liter -1. This exposure was conducted on 3 June 1991.
During the exposure period the temperature within the exposure chamber was measured by a mercury thermometer located at the animals' breathing zone whilst the relative humidity was monitored using wet/dry bulb mercury thermometers. The chamber temperature and
relative humidity were measured at 30 min intervals throughout the exposure period.

Analytical verification of test atmosphere concentrations:

yes

Remarks:

Chamber concentrations were measured at regular interval s during the exposure period. The gravimetric method used employed silica gel sorbent sampling tubes.

Duration of exposure:

4 h

Remarks on duration:

As per guideline

Concentrations:

Group 1 (5 males and 5 females): 4.07 mg/ litre -1
Group 2 (5 males and 5 females): 7.53 mg/ litre -1

No. of animals per sex per dose:

5 male and 5 female.

Control animals:

not specified

Details on study design:

Atmospheric Characterisation

Chamber Concentration:
Chamber concentrations were measured at regular interval s during the exposure period. The gravimetric method used employed silica gel sorbent sampling tubes. The input side of the tube was positioned and temporarily sealed in a port in the exposure chamber at the animals' breathing zone. Chamber air was drawn through the sorbent tube at a measured rate of 1.0 litre.min -1 using a vacuum pump. The air flow during each sample was controlled by a flow meter and timed for a suitable period.
Each sorbent tube was weighed before and after sampling in order t o calculate by difference the weight o f material collected. The total chamber concentration was estimated by further calculation using the sample air volume. The background moisture content was determined t o be 0.27 and 0.11 mg.litre -1 for Groups 1 and 2 respectively and these values were subtracted from the above derived chamber concentration.

Nominal Chamber Concentration
The nominal chamber concentration was estimated using the following equation:

Nominal concentration = Weight of material used (mg) / Total air flow through chamber (litres)

Particle size distribution:
The particle size distribution o f the dispersed material inside the exposure chamber was estimated twice during each exposure period using a Marple (Model 296) Cascade Impactor (Anderson Samplers Inc., Atlanta, Georgia, USA). This device consisted of an in line sampler and a series of impaction stages capable of fractionating the aerosol into the size range 0.25->10 µm. The device was positioned on the chamber at the animal's breathing zone chamber air sampled, using a vacuum pump, at a constant rate o f 2.0 1itres.rnina1 for a recorded time period.
The implication substrates (for each stage) and back up sorbent tube were weighed before and after sampling and the weight increase on each substrate determined to be the mass of particles in the size range o f that impactor stage. From the results obtained, the total weight of particles collected on the impaction stages and back up sorbent tube were summed and the percent particle mass in each size range calculated. The data are presented as showing the percent of particle mass smaller that the stated aerodynamic particle diameter, and the respirable mass fraction determined from the particle size distribution.

Observation on the animals

Clinical signs
A17 the rats were observed for clinical signs at frequent intervals throughout the exposure period, for the first 1 h post dosing and thereafter at least once (normally twice) daily during the subsequent 14 day observation period. The onset, intensity and duration of any signs observed were recorded.

Body Weights
All the rats were weighed immediately. before dosing and on Days 2, 3, 4, 7, 10 and 14 post exposure.

Terminal studies

Necropsy examination:
A t the end of the 14 day observation period a11 the animals were sacrificed by an intraperitoneal overdose of pentobarbitone sodium (Expiral, Abbot) and subjected to a macroscopic post mortem examination.
Each rat was examined externally prior to opening the abdominal and thoracic cavities. Any gross lesions observed were recorded in descriptive terms, including location(s) , size (mm) , colour and number. The respiratory tract was subjected to detailed macroscopic examination for signs of irritancy or local toxicity. All organs were examined in situ.

Lung : Body weight ratio:
The lungs o each animal were removed and weighed to allow calculation of lung : body weight ratios.
On completion of the necropsy, the animals' carcasses were disposed of; no tissues were retained.

Preliminary study:

Not specified

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 7.53 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

There were no mortalities during the course of the study.

Clinical signs:

other: An unkempt appearance was noted for all Group 1 animals immediately post dose and one animal (179, Group 2) appeared subdued and showed hunched posture over the first 24 h post dose. Otherwise no other abnormalities were detected following treatment.

Body weight:

There was no effect on body weight profile following exposure to ANOX BF

Gross pathology:

All group 1 animals were seen to have no abnormalities.
Slightly mottled lungs i n all but 2 Group 2 animals and pale/discoloured lungs were noted in one Group 2 male (No. 12) and 1 Group 2 female (No 20). These were deemed to be in accordance with normal background findings i n acute rat studies at IRI and not attributable to treatment.

Other findings:

Lung : body weight ratios were generally within normal limits for all animals, although Group 2 males tended towards the bottom of the expected range.

The chamber air flow rate was monitored continuously and recorded at 30 min intervals throughout the exposure period. The values recorded were a constant 12.0 litre.min^-1, for both groups.

Chamber Atmos~heric Conditions
Concentration
The gravimetric concentration of the test aerosols in the exposure chamber was derived from the mean of several measurements made during the exposure periods. The adjusted overall mean values obtained were 4.07 mg litre^-1 for Group 1 and 7.53 mg. litre^-1 for Group 2.

Particle Size Distribution
Estimation of the particle size distribution revealed that the percentage of particles <3.5 µm for Group 1 was 94.0 and for Group 2 was 93.9.

 

Interpretation of results:

GHS criteria not met

Conclusions:

ANOX BF did not produce any evidence of toxicity in Sprague-Dawley rats following exposure for 4 hours to an atmosphere containing 7.53 mg. liter -1 (measured gravimetrically).

Executive summary:

This study was undertaken to investigate the acute inhalation toxicity (limit test) of ANOX BF  in two groups of rats (5male and 5 female) following a single 4 hour snout only exposure to the test article.

The study was conducted in compliance with OECD (403) guidelines for acute toxicity testing and in accordance with internationally recognised standards of GLP.

Group 1 animals were exposed to a measured gravimetric ^{conce-1ntration} of 4.07 mg. litre ^-1 (87.85 mg. litre^-1 nominal). Group 2 animals were exposed to a measured gravimetric concentration of 7.53. litre^-1 (70.14 mg. litre^-1 nominal). The particle size determination indicated 94.0% and 93.9% of the test aerosol particles to be <3.5 µm for groups 1 and 2 respectively.

There were no mortalities, clinical signs or affects to body weight following exposure to the test article. No abnormalities detected at post mortem observations were considered to be indicative of a reaction to treatment with the test article.

The lung : body weight ratio values were generally all within normal limits.

In conclusion, ANOX BF did not produce any evidence of toxicity in Sprague-Dawley rats following exposure for 4 hours to an atmosphere containing 7.53 mg. litre^-1 (measured gravimetically).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

7.53 mg/m³ air

Quality of whole database:

K1

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

Study period:

February 06 1991 to February 21 1991

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

other: 84/449/EWG, B.3

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

Not specified

Species:

rat

Strain:

Sprague-Dawley

Remarks:

Crl:CD(SD) BR

Sex:

male/female

Details on test animals or test system and environmental conditions:

Body weight (on receipt): Males: 225-250 g
Females: 200-225 g
Age (on receipt): About 7-9 weeks
Supplier: Charles River Italia S.p.A. Via Indipendenza, 11, 22050 - CALCO (Como),
Shipping slip number: 00621 dated Jan. 21, 1991
Acclimation: at least 5 days before the start of the test. Animals were observed daily to ascertain their fitness for the study.
Housing: 1 animals/cage in air-conditioned rooms.
- Temperature: 22 ° C ± 2
- Relative humidity: 55% ± 10
- Air changes: about 20/hour filtered on HEPA 99.97%.
- HEPA 99.97%
- Light: 12 hour cycle (7 a.m. - 7 p.m.)
- Cage size: grill cages cm 40.5x38.5x18h with stainless steel feeder. The waste that
dropped through the grill bottom on removable paper was periodically disposed of.
Animal identification: By nicks or tipping's of the outer ear. Cage card gave experiment number, dosage group, sex and date of administration.
Diet: GLP 4RF21 pelleted diet produced by Charles River Italia' s feed licensee Yucedola
S.r.l., Settimo Yilanese. The declared contents, on the label, on dry matter basis (moisture 12%), were:
crude protein 13.50%
crude fat 3.00%
crude fiber 6.00%
ash 7.00%
The diet was supplemented by the Producer with vitamins and trace elements. The Producer supplies a certificate of anal:-sis for nutrients and contaminants, the levels of which are within the limits proposed by EPA-TSC-4 (44FR:4053-44093, July 26, 1979). RBM has the animal feed re-analyzed at Least twice a year for bacterial contamination. The diet was available "ad libitum" to the animals.
Water: from the municipal water main system. water is filtered and distributed "act· libitum" to the animals by an automatic valve system.
Periodically drinking water is analyzed for microbiologic count. heavy metals, other contaminants (e.g. solvents, pesticides) and other chemical and physical characteristics. The acceptable limits of quality of the drinking water were those defined in EEC Directive 80/778.

Contaminants that might interfere with the objectives of the study were not expected to be present in diet or drinking water.

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

Dermal exposure:
Preparation of animals skin: Approximately 24 hours before the test, fur was clipped from the dorsal and ventral area of the trunk of the test animals. Care was taken to avoid abrading the skin which could alter its permeability.
An area of about 6x5 cm of the body dorsal surface was cleared for the application of the test article. This area corresponding to about 10% of the total body surface.

Administration of the test article:
By uniform application onto the cleared area. The treated area was covered with a porous gauze dressing fixed to the skin with hypoallergenic non-irritating tape (Micropore, 3M Italia, Milano).
The test side was further covered in a suitable manner in order to ensure that the animals could not ingest the test substance. At the end of the exposure period the residual substance was wiped off.

Duration of exposure:

24 hours exposure

Doses:

single does of 2000 mg/Kg body weight

No. of animals per sex per dose:

5 males + 5 females/group

Control animals:

not specified

Details on study design:

Study design
Administration route: epidermal
Reason for selection of administration route : Possible accidental exposure for humans.
Administration frequency: Single.
Exposure period: About 24 hours.

Experimental design: One group of 5 rats/sex was administered a single dose of 2000 mg/Kg of the test article.
Individual dosages were based on body weight taken just before treatment.

Observation period: 14 days after the administration.
Observation of clinical signs: On exposure day at 30 minutes, 2, 4 and 6 hours on the following 14 days the animals were observed twice a day for behavioral and clinical changes.

Observation of mortality: Twice a day, early in the morning and late in the afternoon.

Body weight: Once pre-trial, on the administration day (day 1 to calculate the administration volumes) and on days 8 and 15.

Gross pathology: On all the animals (fasted overnight) at the termination of the 14 days post exposure observation period.
Animals were killed by excision of the femoral arteries, after i.p. anesthesia with 5% sodium pentobarbital.

Histology: Since no changes were found at necropsy, histological examination was not performed.

Statistics:

Since no mortality occurred at the limit dose of 2000 mg/Kg, the LD50 was not calculated.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed in animals treated at 2000 mg/Kg (limit dose) during the post-treatment observation period.

Clinical signs:

other: The rats did not show any clinical sign or change in behavior and no reaction was observed at the application site.

Gross pathology:

No changes were observed in the animals killed at the end of the study.

Interpretation of results:

GHS criteria not met

Conclusions:

The test article ANOX - BF, when administered by dermal route to rat, under the conditions adopted in this experiment, did not cause mortality nor toxic effects at the limit dose of 2000 mg/Kg.
The LD50 by dermal route is higher than 2000 mg/Kg

Executive summary:

A acute dermal toxicity study was carried out in rats treated with test article ANOX – BF,  according to test guideline 84/449/EWG, B.3, in compliance with GLP.

Experimental data obtained from a toxicity study in which Sprague Dawley Crl:CD (SD) BR rats ( 5 males and 5 females/group) received a single dermal administration of the test article ANOX – BF at the dose of 2000 mg/Kg (24 hour exposure) are provided.

The animals were observed for 14 days after the patch removal.

No deaths occurred as a result of treatment.

No clinical signs or behavioural alterations were observed in any animal.

At the application site, no dermal reactions were detected.

No significant changes in the body weight gain were observed.

No changes were found at the gross pathology examination performed at the end of the observation period.

In conclusion, the test article ANOX - BF, when administered by dermal route to rat, under the conditions adopted in this experiment, did not cause mortality nor toxic effects at the limit dose of 2000 mg/Kg.

The LD50 by dermal route is higher than 2000 mg/Kg

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

K1

Additional information

Acute Oral Toxicity
A acute oral toxicity study was carried out in rats treated with test article ANOX – BF,  according to test guideline 84/449/EWG, B.1, in compliance with GLP.

The test article was suspended in 0,5% methylcellulose 400 cps water

Solution and administered to rats at the volume of 20 ml/kg.

All rats were treated after a 16 hrs fasting period. The animals were weighed on days 1, 3, 8 and 14; clinically observed for 14 days following the treatment and killed at the end of the study by excision of the femoral arteries after having been completely anesthetized with an i.p. injection of 5% sodium pentobarbital . A thorough autoptic  examination was performed in animals killed at the end of the observation period .

No animals died during the observation period.

Two male and three female treated rats only showed transient piloerection 2 hours after treatment.

The body weight gain appeared unaffected by treatment .

At the gross pathology examination performed on rats killed at the end of the observation period no changes were noted.

In conclusion ANOX - BF, when administered by oral rout to rat, under' the conditions adopted in this study, did not cause overt signs of toxicity- and showed an LD50 higher than 5000 mg/kg.

 

Acute Dermal Toxicity
A acute dermal toxicity study was carried out in rats treated with test article ANOX – BF,  according to test guideline 84/449/EWG, B.3, in compliance with GLP.

Experimental data obtained from a toxicity study in which Sprague Dawley Crl:CD (SD) BR rats ( 5 males and 5 females/group) received a single dermal administration of the test article ANOX – BF at the dose of 2000 mg/Kg (24 hour exposure) are provided.

The animals were observed for 14 days after the patch removal.

No deaths occurred as a result of treatment.

No clinical signs or behavioural alterations were observed in any animal.

At the application site, no dermal reactions were detected.

No significant changes in the body weight gain were observed.

No changes were found at the gross pathology examination performed at the end of the observation period.

In conclusion, the test article ANOX - BF, when administered by dermal route to rat, under the conditions adopted in this experiment, did not cause mortality nor toxic effects at the limit dose of 2000 mg/Kg.

The LD50 by dermal route is higher than 2000 mg/Kg

 

Acute Inhalation Toxicity

This study was undertaken to investigate the acute inhalation toxicity (limit test) of ANOX BF  in two groups of rats (5male and 5 female) following a single 4 hour snout only exposure to the test article.

The study was conducted in compliance with OECD (403) guidelines for acute toxicity testing and in accordance with internationally recognised standards of GLP.

Group 1 animals were exposed to a measured gravimetric conce-1ntration of 4.07 mg. litre -1 (87.85 mg. litre-1 nominal). Group 2 animals were exposed to a measured gravimetric concentration of 7.53. litre-1 (70.14 mg. litre-1 nominal). The particle size determination indicated 94.0% and 93.9% of the test aerosol particles to be <3.5 µm for groups 1 and 2 respectively.

There were no mortalities, clinical signs or affects to body weight following exposure to the test article. No abnormalities detected at post mortem observations were considered to be indicative of a reaction to treatment with the test article.

The lung : body weight ratio values were generally all within normal limits.

In conclusion, ANOX BF did not produce any evidence of toxicity in Sprague-Dawley rats following exposure for 4 hours to an atmosphere containing 7.53 mg. litre-1 (measured gravimetically).

Justification for classification or non-classification

Based on the studies conducted the substance does not meet the criteria specified in Regulation 1272/2008 for classification for acute toxicity.