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Description of key information

In an acute oral toxicity study with rats, performed according to OECD 401 test guideline and GLP principles, an LD50 >2000 mg/kg bw was determined.
In a supporting acute oral toxicity study with rats, performed equivalent to OECD 401 test guideline, an LD50 >15000 mg/kg bw was determined.
In an acute dermal toxicity study with rats, performed according to OECD 402 test guideline and GLP principles, an LD50 >2000 mg/kg bw was determined.
In an acute inhalation toxicity study with rats, performed according to OECD 403 test guideline and GLP principles, an LC50 >5 mg/L bw was determined.

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:
10 May, 1995 - 24 May, 1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Version / remarks:
(1987)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
(1992)
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: Wistar Crl: (WI) BR
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River, Germany.
- Age at study initiation: Approx. 11 weeks
- Weight at study initiation: males: 386 - 438 g; females: 255 - 276
- Fasting period before study: Food was withheld overnight prior to dosing until approx. 3-4 hours after administration of the test substance.
- Housing: Group housing of 5 animals per cage in labelled polycarbonate cages.
- Diet: Free access to standard pelleted laboratory animal diet (Carfil Quality BVBA, Oud-Turnhout, Belgium).
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): 50
- Air changes (per hr): approx 15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
propylene glycol
Details on oral exposure:
VEHICLE
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed at NOTOX and on test substance data supplied by the sponsor.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg body weight.
Frequency: single dosage, on Day 1.

DOSAGE PREPARATION: The formulation (w/w) was prepared immediately prior to dosing. Adjustment was made for specific gravity of vehicle (1.036). Homogeneity was accomplished to a visually acceptable level.
Doses:
2000 mg/kg body weight


No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Viability: Twice daily
Body weights: Days 1 (pre-administration), 8 and 15.
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
- Necropsy of survivors performed: All animals surviving to the end of the observation period (day 15) were sacrificed by oxygen/carbon dioxide asphyxiation. All animals assigned to the study were subjected to necropsy and descriptions of all macroscopic abnormalities recorded.
- Other examinations performed: none.
Statistics:
No statistical analysis was performed.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No animals died.
Clinical signs:
Uncoordinated movements were observed in two males on day 1 and hunched posture and piloerection were observed in one female on days 3 and 4.
Body weight:
The body weight gain shown by the animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.
Gross pathology:
Macroscopic post mortem examination of the animals at termination did not reveal any abnormalities.
Other findings:
None.
Interpretation of results:
other: Not classified
Remarks:
According to Regulation (EC) No. 1272/2008 and its amendments.
Conclusions:
In an acute oral toxicity study with male and female rats, performed according to OECD 401 test guideline and GLP principles, an LD50 >2000 mg/kg bw was determined.
Executive summary:

Alcamizer 5 was tested in an acute oral toxicity study with male and female rats, performed according to OECD 401 test guideline and GLP principles.

No mortality occurred. Uncoordinated movements were observed in two males on day 1 and hunched posture and piloerection were observed in one female on days 3 and 4. The body weight gain shown by the animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain. No abnormalities were found at macroscopic post mortem examination of the animals. Based on the results, an LD50 >2000 mg/kg bw was determined. Alcamizer 5 does not have to be classified and has no obligatory labelling requirement for acute oral toxicity according to Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
The study has klimisch code 1.

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:
10 August, 1995 - 24 August, 1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Version / remarks:
(1981)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: Crl:[WI]WU BR
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga, Sulzfeld, Germany
- Age at study initiation: appr. 9 weeks
- Weight at study initiation: 305 - 327 g (males); 193 - 201 g (females)
- Fasting period before study: no
- Housing: Group housing of 5 animals/sex in suspended stainless steel cages
- Diet: free access to cereal-based rodent diet (SDS Special Diets Services, Witham, England) (no access to food during exposure)
- Water: free access to tap water (no access to water during exposure)
- Acclimation period: total 17 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.5 - 24.0
- Humidity (%): 63 - 89
- Air changes (per hr): approx. 10
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
inhalation: dust
Type of inhalation exposure:
nose only
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Nose-only inhalation chamber, a modification of the chamber manufactured by ADG Developments Ltd., Codicote, Hitchin, Herts, SG4 8UB, UK.
- Method of holding animals in test chamber: the animals were secured in plastic animal holders
- Exposure chamber volume: 150 L
- Rate of air: mean amount 104 L/min
- System of generating particulates/aerosols: The test atmosphere was generated by passing the test material using a dry material helix feeder to a jet mill. The jet mill was operated with dry pressurized air (less than 1% humidity); the test material was delivered using a slip stream of airconditioned room air and pressurized air. This stream accounted for about 55% of the total amount of air. The generated aerosol was passed to the inlet of the exposure unit and was directed downward through the mixing chambers towards the animal noses. At the bottom of the unit the test atmosphere was exhausted.
- Method of particle size determination: Twice during exposure using a 10-stage Anderson cascade impactor with the largest cut-off size of 32 µm. Due to a technical failure these measurements could not be used to determine the particle size distribution. The particle size distribution measurement, however, had also been carried out the day before exposure during a test run at exactly the same settings as during exposure.
- Treatment of exhaust air: no data
- Temperature, humidity in air chamber: 20. 5 - 21.0°C; 41 - 58%

TEST ATMOSPHERE
- Brief description of analytical method used: The actual concentration in the test atmosphere was determined twice each hour by gravimetric analysis. The nominal concentration was determined by dividing the total amount of test material used by the total volume of air passed through the exposure unit.
- Samples taken from breathing zone: Representative samples were obtained by passing test atmosphere samples at ca. 5 L/min through fiber glass filters. Before sampling the filters were weighed; immediately after sampling the filters were weighed again. The actual concentration was calculated by dividing the amount of test material present on each filter by the volume of the test atmosphere sample taken.

TEST ATMOSPHERE
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): 1.9 µm / 2.2 µm

CLASS METHOD
- Rationale for the selection of the starting concentration: Based on the cut off concentration values specified in the UN and EC classification guidelines.
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
5 mg/L
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Behaviour, clinical signs and mortality: The rats were visually inspected just before exposure, for reactions to treatment during the exposure, shortly after exposure, and at least once daily during the observation period.
Body weights: Body weights were recorded just prior to exposure (day 0), and on days 7 and 14.
- Necropsy of survivors performed: At the end of the 14 day observation period, all rats were killed by exsanguination from the abdominal aorta under ether anaesthesia. All rats were necropsied and examined for gross pathological changes.
Statistics:
No statistical analysis was performed (the method used was not intended to calculate a LC50 value).
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5 mg/L air (analytical)
Based on:
test mat.
Exp. duration:
4 h
Mortality:
No animals died.
Clinical signs:
other: Slight visually decreased breathing rate was observed in all rats ca. 1.5, 2.5 and 3.5 hours after the start of the exposure. Two males and one female felt cold upon touching shortly after exposure. Dirty fur of the head was seen in 2 females until day 7
Body weight:
Slightly reduced mean body weight gain was generally seen in all rats 7 days after exposure. Normal body weight gain was observed in males at the end of the observation period, whereas body weight gain remained low in 3 females.
Gross pathology:
No abnormalities were observed at necropsy, except for one female that showed sparsely haired abdomen.
Other findings:
None.

The mean actual concentration of the substance during the exposure based on gravimetric analyses was 5.16 ± 0.48 mg/L.

The nominal concentration was calculated to be 14.7 mg/L indicating a generation efficiency of ca. 35%.

The particle size distribution: it was shown that 91.2% of the particles present at the animals' breathing zone were of the respitable size range since they were smaller than or equal to 5.0 µm.

MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): 1.9 µm / 2.2 µm

Interpretation of results:
other: Not classified.
Remarks:
According to Regulation (EC) No. 1272/2008 and its amendments.
Conclusions:
In an acute inhalation toxicity study with male and female rats, performed according to OECD 403 test guideline and GLP principles, an LC50 >5 mg/L was determined for ALCAMIZER 5.




Executive summary:

Alcamizer 5 was tested in an acute inhalation toxicity study with nose-only exposure with male and female rats, performed according to OECD 403 test guideline and GLP principles.

No mortality occurred. Slight visually decreased breathing rate was observed in all rats ca. 1.5, 2.5 and 3.5 hours after the start of the exposure. Two males and one female felt cold upon touching shortly after exposure. Dirty fur of the head was seen in 2 females until day 7 of the 14 day observation period. One female additionally showed alopecic areas on the fur on days 7-14. Slightly reduced mean body weight gain was generally seen in all rats 7 days after exposure. Normal body weight gain was observed in males at the end of the observation period, whereas body weight gain remained low in 3 females. No abnormalities were observed at necropsy, except for one female that showed sparsely haired abdomen.

Based on the results, an LC50 >5 mg/L was determined. Alcamizer 5 does not have to be classified and has no obligatory labelling requirement for acute inhalation toxicity according to Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
Value:
5 000 mg/m³
Quality of whole database:
The study has klimisch code 1.

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:
12 April, 1995 - 26 April, 1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
(1987)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
(1992)
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: Wistar Crl: (WI) BR
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River, Germany
- Age at study initiation: Approx. 8 weeks
- Weight at study initiation: males: 288 - 337 g; females: 206 - 229 g
- Housing: Individually housed in labelled polycarbonate cages.
- Diet: Free access to standard pelleted laboratory animal diet (Carfil Quality BVBA, Oud-Turnhout, Belgium)
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): 50
- Air changes (per hr): approx. 15
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
occlusive
Vehicle:
corn oil
Details on dermal exposure:
One day before exposure (Day -1) an area of approximately 5x7 cm on the back of the animal was clipped.

The formulation was applied in an area of approx. 25 cm^2 (5x5 cm) for males and 18 cm^2 (3.5x5 cm) for females by application on a gauze patch fixed successively to aluminium foil and flexible bandage, with drops of petrolatum.

Frequency: Single dosage, on Day 1.

Washing: Following application, dressings were removed and the skin cleaned of residual test substance using a tissue moistened with tap water.
Duration of exposure:
24 h
Doses:
2000 mg/kg
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
VEHICLE
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed at NOTOX and on test substance data supplied by the sponsor.

Dose volume: 10 mL/kg

DOSAGE PREPARATION: The formulation (w/w) was prepared immediately prior to dosing. Adjustment was made for specific gravity of vehicle (0.92). Homogeneity was accomplished to a visually acceptable level.

Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Viability: Twice daily
Body weights: Days 1 (pre-administration), 8 and 15
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
- Necropsy of survivors performed: All animals surviving to the end of the observation period (day 15) were sacrificed by oxygen/carbon dioxide asphyxiation. All animals assigned to the study were subjected to necropsy and descriptions of all macroscopic abnormalities recorded.
- Other examinations performed: none.
Statistics:
None.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
No animals died.
Clinical signs:
No clinical signs were observed.
Body weight:
The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not toxicologically significant.
Gross pathology:
Macroscopic post mortem examination of the animals at termination did not reveal any abnormalities.
Other findings:
Erythema, scales and scabs were seen in the treated skin-area among four females during the observation period.

One female had lost the patch within 24 hours. The results from this animal were excluded from interpretation. 

This did not affect study integrity.

Interpretation of results:
other: Not classified.
Remarks:
According to Regulation (EC) No. 1272/2008 and its amendments.
Conclusions:
In an acute dermal toxicity study with rats, performed according to OECD 402 test guideline and GLP principles, an LD50 >2000 mg/kg bw was determined.
Executive summary:

Alcamizer 5 was tested in an acute dermal toxicity study with male and female rats, performed according to OECD 402 test guideline and GLP principles.

No mortality occurred. The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not toxicologically significant. No abnormalities were found at macroscopic post mortem examination of the animals. Erythema, scales and scabs were seen in the treated skin-area among four females during the

observation period. Based on the results, an LD50 >2000 mg/kg bw was determined. Alcamizer 5 does not have to be classified and has no obligatory labelling requirement for acute dermal toxicity according to Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
The study has klimisch code 1.

Additional information

Acute oral:

Alcamizer 5 was tested in an acute oral toxicity study with male and female rats, performed according to OECD 401 test guideline and GLP principles.

No mortality occurred. Uncoordinated movements were observed in two males on day 1 and hunched posture and piloerection were observed in one female on days 3 and 4. The body weight gain shown by the animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain. No abnormalities were found at macroscopic post mortem examination of the animals. Based on the results, an LD50 >2000 mg/kg bw was determined.

Alcamizer 5 was tested in an acute oral toxicity study with male and female rats, performed equivalent to OECD 401 test guideline.

No mortality occurred. Decreased spontaneous activity, hunchback, ptosis and piloerection was observed among the animals 10 minutes after administration of 3800, 7500 and 15000 mg/kg. The symptoms, except for hunchback, disappeared until 6 hours after administration. Furthermore, cuddy feces and periproctal soil were observed in some males receiving 3800 mg/kg or more and in some females receiving 1900 mg/kg or more on the day of administration. The day after administration only pale brown feces was observed in some animals receiving 3800 mg/kg or more. No abnormalities were observed from day 2, except for the attachment of blood-like substance around the eyelid and nose and emaciation in one female at 1900 mg/kg from day 11. Body weight gained well and dose-dependent change was not observed except the body weight decreased of only one female in 1900 and 15000 mg/kg group on day 14 or 10. No abnormalities were found at macroscopic post mortem examination of the animals, except for the red color change of submandibular lymph node in one female receiving 1900 mg/kg. Based on the results, an LD50 >15000 mg/kg bw was determined.

Acute inhalation:

Alcamizer 5 was tested in an acute inhalation toxicity study with nose-only exposure with male and female rats, performed according to OECD 403 test guideline and GLP principles.

No mortality occurred. Slight visually decreased breathing rate was observed in all rats ca. 1.5, 2.5 and 3.5 hours after the start of the exposure. Two males and one female felt cold upon touching shortly after exposure. Dirty fur of the head was seen in 2 females until day 7 of the 14 day observation period. One female additionally showed alopecic areas on the fur on days 7-14. Slightly reduced mean body weight gain was generally seen in all rats 7 days after exposure. Normal body weight gain was observed in males at the end of the observation period, whereas body weight gain remained low in 3 females. No abnormalities were observed at necropsy, except for one female that showed sparsely haired abdomen.

Based on the results, an LC50 >5 mg/L was determined.

Acute dermal:

Alcamizer 5 was tested in an acute dermal toxicity study with male and female rats, performed according to OECD 402 test guideline and GLP principles.

No mortality occurred. The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not toxicologically significant. No abnormalities were found at macroscopic post mortem examination of the animals. Erythema, scales and scabs were seen in the treated skin-area among four females during the

observation period. Based on the results, an LD50 >2000 mg/kg bw was determined.


Justification for selection of acute toxicity – oral endpoint
One acute oral study is available according to OECD guideline and GLP principles.

Justification for selection of acute toxicity – inhalation endpoint
One acute inhalation study is available.

Justification for selection of acute toxicity – dermal endpoint
One acute dermal study is available.

Justification for classification or non-classification

Based on the above study results, Alcamizer 5 does not have to be classified and has no obligatory labelling requirement for acute oral, acute dermal and acute inhalation toxicity according to Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments).