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Description of key information

Testing for acute oral toxicity in rats revealed a reliable LD50 to be 562 mg/kg bw. Testing for acute dermal toxicity a median lethal dose of 1500 mg/kg bw in rats was found. No mortality was observed when rats were exposed for 8 hours to saturated vapour of the submission substance (i.e. nominal concentration 3.1 mg/l).

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:
1986
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Basic data given: comparable to guidelines/standards
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Dr. Thomae, Bibrach, D
- Mean weight at study initiation: males 185 g, females 174 g (+- 20%)
- Fasting period before study: 16 h
- Housing: 5 per cage in stainless steel wire mesh cages, Typ DK-III
- Diet (e.g. ad libitum): Kliba-Labordiet, Klingenthalmuehle AG, Kaiseraugst, CH; ad libitum
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period: at least 1 week

ENVIRONMENTAL CONDITIONS in fully air-conditioned rooms
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 3.83 - 8.25 % (w/v)

MAXIMUM DOSE VOLUME APPLIED: 10 mL
Doses:
383, 562 and 825 mg/kg bw
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: observations: several times on the day of administration and at least once each workday; check for moribund and dead animals twice each workday and once on holidays; weighing on day 0, 2, 7 and 13
- Necropsy of survivors performed: yes
Sex:
male/female
Dose descriptor:
LD50
Effect level:
ca. 562 mg/kg bw
Based on:
test mat.
Remarks on result:
other: Within the 14 days observation period no animals died at a dose level of 383 mg/kg bw. At 562 mg/kg bw 2/5 males and 2/5 females died. Application of 825 mg/kg bw resulted in death of all animals. All deaths occurred within 1-2 days after administration
Mortality:
383 mg/kg bw: 0 males and 0 females died
562 mg/kg bw: 2 males and 2 females died
825 mg/kg bw: all animals died
All animals that died were dead within 1-2 days after administration
Clinical signs:
383 mg/kg bw: no clinical signs observed
562 mg/kg bw: dyspnea, apathy, abnormal position (only males), staggering, atonia, paresis, spastic gait (only females), piloerection, exsiccosis and poor general state; reversible in surviving animals within at least two days after administration
825 mg/kg bw: in addition to the already cited signs, ophisthotonus in males; observed in animals in agony
Body weight:
Mean weights on day 0, 2, 7 and 13 in grams
383 mg/kg bw
males: 189 - 209 - 242 - 272
females: 172 - 187 - 197 - 207
562 mg/kg bw
males: 189 - 202 - 236 - 269
females: 180 - 175 - 201 - 219
Normal body weight gain observed in the survivors.
Gross pathology:
Animals-that died (male and female):
General congestive hyperemia
Stomach: severe redness of the glandular stomach
Small intestines, cecum: severe redness, filled with hematinised contents.

Sacrificed animals (male and female):
no abnormalities detected.
Interpretation of results:
harmful
Remarks:
Migrated information Acute toxicity category 4 Criteria used for interpretation of results: EU
Conclusions:
After single application of either 383, 562 or 825 mg test substance per kg into male and female lethality could be observed during the 14 day observation period, resulting in a LD50 of approx. 562 mg/kg bw.
Executive summary:

Acute oral toxicity was investigated using male and female Wistar rats in a test similar to acute standard method (i.e. OECD TG 401). The test substance was administered by gavage at doses of 383, 562 and 825 mg/kg bw to 3 groups of 10 animals (5animals/sex). Clinical signs, e.g. dyspnea, apathy, atonia etc., were observed starting from the mid-dose group. Within the 14 days observation period 2 males and 2 females died from the mid-dose group, whereas all animals died from the high-dose group. Having this results a median lethal dose (LD50) of approximately 562 mg/kg bw was identified.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
562 mg/kg bw
Quality of whole database:
Various studies for the submission substance were presented which show comparable results. The quality of the datbase is therefore high.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1958
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline with acceptable restrictions (mostly due to limited documentation; nominal determination of the test atmosphere; strongly reduced turnover of the test atmosphere (maximum 20 L/h))
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Version / remarks:
adopted 1981, Annex
Deviations:
yes
Remarks:
determination of the test atmosphere by reweighing of a test substance column
GLP compliance:
no
Test type:
other: IHT (Inhalation hazard test)
Limit test:
yes
Species:
rat
Strain:
not specified
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Mean weight at study initiation: 126 g (animals of both sexes and both trials combined)

ENVIRONMENTAL CONDITIONS
not reported
Route of administration:
inhalation: vapour
Type of inhalation exposure:
whole body
Vehicle:
air
Details on inhalation exposure:
Rats were exposed for 8 h to a vapour saturated atmosphere.
Vapour was generated by bubbling dry air (no CO2) through the liquid substance column (volume ca. 120 ml) above a fritted glass disc in a glass cylinder. This glass cylinder was placed in an oil bath with a temperature of 50°C. Temperature in the exposure chamber was 20°C.
Due to the increasing viscosity of the test substance during the exposure period, the normally used turnover of 200 L/h dropped down to maximum 20 L/h which was about half of the needed amount of fresh air (36 L/h per 6 animals).
Analytical verification of test atmosphere concentrations:
no
Duration of exposure:
8 h
Concentrations:
3.1 mg/L nominal
No. of animals per sex per dose:
3 animals of each sex
Control animals:
no
Details on study design:
- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: Groupwise weighing was done on days 0, 4, 8, 11 and 15. Detailed clinical observations were performed several times at the day of exposure and daily with the exception of weekends and holidays afterwards.
- Necropsy of survivors performed: no
Sex:
female
Dose descriptor:
other: IHT
Effect level:
3.1 mg/L air
Based on:
test mat.
Remarks:
nominal
Exp. duration:
8 h
Remarks on result:
other: 0/6 animals died during exposure to the volatile parts of the test item for 8 h
Mortality:
0/6 animals died
Clinical signs:
other: After 40 min of exposure eyelid closure and ruffled fur, after 5 h gasping. On the day after application free from symptoms.
Body weight:
Constant body weight gain during the observation period.
Gross pathology:
not performed.
Conclusions:
Testing for acute toxicity after exposure through inhalation a limit test equivalent to the one set out in OECD TG 403 was performed. None of the six animals (male and female rats) exposed for 8 hours to saturated vapour containing 3.1 mg test item per l air (nominal determination) died during the 14 day observation period. Having these results one could state that under the conditions tested the median lethal dose of the test item is > 3.1 mg/l.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating conc.
Value:
3 100 mg/m³
Quality of whole database:
Based on the tonnage driven data requirements two routes of exposure have to be considered for acute toxicity. Data on oral and dermal acute toxicity are available. The toxicity study after inhalation exposure is given as supporting data.

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:
1978
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Comparable to guideline study with acceptable restrictions (mostly due to reduced reporting in times before GLP, e.g. no single animal data presented, results of preliminary test not reported, lacking information on animal husbandry, occlusive conditions)
Reason / purpose for cross-reference:
reference to same study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
mostly due to reduced reporting in times before GLP, e.g. no single animal data presented, results of preliminary test not reported, lacking information on animal husbandry, occlusive conditions
GLP compliance:
no
Remarks:
study performed before GLP statement
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Oncins, IFFA CREDO, France
- Weight at study initiation: 120-150 g

ENVIRONMENTAL CONDITIONS
no further details
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: 6cm x 6cm
- Type of wrap if used: aluminium foil and adhesive tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): with lukewarm soap water
- Time after start of exposure: 24 h (directly after patch removal)

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.85 ml/kg
- Other: as the test material was solid at 24 °C as well as in between 32 and 34°C, the test material was heated to 37 °C, became fluid and was administered pure to the skin of test animals
TEST SITE
- Area of exposure:
- % coverage:
- Type of wrap if used:

REMOVAL OF TEST SUBSTANCE
- Washing (if done):
- Time after start of exposure:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Constant volume or concentration used: yes/no
- For solids, paste formed: yes/no

VEHICLE
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Lot/batch no. (if required):
- Purity:
Duration of exposure:
24 hours
Doses:
740, 1110, 1670, and 2500 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
other: one group of vehicle treated animals were kept in order to determine normal body weight development.
Details on study design:
- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: every 5th day (i.e. study day 0, 5, 10 and 15) for vehicle treated groups and animals treated with 740 mg/kg bw (which is the highest dose which did not produce deaths)
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight and gross pathology
Statistics:
LD50 value was calculated using the method of Litchfield and Wilcoxon.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
1 500 mg/kg bw
Based on:
test mat.
95% CL:
1 160 - 1 930
Remarks on result:
other: Within the 14 days observation period no animals died after single application of 740 mg/kg bw. 1/10, 8/10 and 9/10 animals died using 1110, 1670 and 2500 mg/kg bw respectively. Deaths occurred within 1 to 5 days post application.
Mortality:
740 mg/kg bw: 0 males and 0 females died
1110 mg/kg bw: 1/10 animals died, death occurred within 5 days after administration
1670 mg/kg bw: 8/10 animals died, death occurred within 2 to 5 days after administration
2500 mg/kg bw: 9/10 animals; death occurred 1 to 4 days after administration )
All animals that died were dead within 1 to 5 days after administration
Clinical signs:
740 and 1110 mg/kg bw: Less severe effects on motoric activity as described at the highest dose level.
1670 mg/kg bw: same symptoms as described in the high dose group with cachexia being obvious at treatment days 1 to 5 within the surviving animals.
2500 mg/kg bw: Agitation within the 1st hour post administration. One day after start of exposure animals displaying prostration with tremor and convulsions were seen. Moreover a decrease of body temperature and decline in breathing depth with ptosis was noted.
Body weight:
Treated animals showed slightly decreased body weight gains compared to untreated control animals. The difference being significant at 5 days post administration. However at the end of the study differences between treated and control animals were no more visible.
Gross pathology:
740 mg/kg bw: In 7 animals necrosis was obvious 7days after test item administration. In the other animals eschar formation was seen 7 days after dose administration and scarring began on day 14 after start of exposure.
1110 mg/kg bw: In 3 animals necrosis was obvious 2 hours after the patch removal. Eschar formation in the other animals started at day 7 after test item administration. Just before the end of the observation period scarring could be observed.
1670 mg/kg bw: in the surviviing animals (i.e. 2/10) beginning at three days after administration, necrosis was obvious. The edges were indured and persistet up until the end of the observation period.
2500 mg/kg bw: the only surviving animal showed induration of skin which began to recover from day 4 until the end of the observation period
  Bodyweight development: Bodyweights given as group mean [g]+/- standard deviation after "n" days in observation
 Dose level n=0   n= 5  n=10  n=15
 Control (untreated)  143 +/- 4.2 172 +/- 7.2   198.2 +/- 20.4  217 +/-27.8
 740 mg/kg bw of test item  143.5 +/-2.1 t = 0.34 n.s. 160.3 +/-7.3 t = 3.77 s.s.   194.5 +/-11.6 t = 0.49 n.s. 210.5 +/-19.5 t = 0.6 n.s. 

n.s. difference to control group not statistically significant

s.s. difference to control group statistically significant

Interpretation of results:
harmful
Remarks:
Migrated information Acute toxicity category 4 Criteria used for interpretation of results: EU
Conclusions:
After single dermal application of either 740, 1110, 1670 or 2500 mg test substance per kg onto the skin of male and female rats lethality could be observed during the 14 day observation period, resulting in a LD50 of 1500 mg/kg bw.
Executive summary:

Acute dermal toxicity was investigated using male and female Sprague-Dawley rats in a test similar to acute standard method (i.e. OECD TG 402, non GLP). The test substance was administered to the skin for 24 hours under occlusive conditions at doses of 740, 1110, 1670 and 2500 mg/kg bw to 4 groups of 10 animals (5animals/sex).

Clinical signs (dyspnea, agitation, apathy, atonia (with tremor and convulsions), hypothermia, cachexia) were observed starting from the mid-dose group. Within the 14 days observation period no animals died after single application of 740 mg/kg bw. 1/10, 8/10 and 9/10 animals died using 1110, 1670 and 2500 mg/kg bw respectively. Deaths occurred within 1 to 5 days post application. Having this results a median lethal dose (LD50) of 1500 mg/kg bw was identified for acute dermal toxicity.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
1 500 mg/kg bw
Quality of whole database:
Study was performed pre-GLP but basic standards were given, thus the study has a Klimisch score 2.

Additional information

Acute oral toxicity was investigated using male and female Wistar rats in a test similar to acute standard method (i.e. OECD TG 401). The test substance was administered by gavage at doses of 383, 562 and 825 mg/kg bw to 3 groups of 10 animals (5animals/sex). Clinical signs (dyspnea, apathy, abnormal position (only males), staggering, atonia, paresis, spastic gait (only females), piloerection, exsiccosis and poor general state; reversible in surviving animals within at least two days after administration; at highest dose in addition: ophisthotonus in males; observed in animals in agony) were observed starting from the mid-dose group. Within the 14 days observation period 2 males and 2 females died from the mid-dose group, whereas all animals died from the high-dose group. Having this results a median lethal dose (LD50) of approximately 562 mg/kg bw was identified.

Several other studies conducted with the submission substance using rats revealed median lethal doses in the range of approx. 970 and 1300 mg/kg bw for male/female animals (supporting studies reliability 1 or 2). Another study stated an approximate lethal dose (i.e. lowest dose administered causing death) in male rats to be 1500 mg/kg bw.

Based on the results of all these studies, the test substance is considered as harmful if swallowed.

Acute dermal toxicity was investigated using male and female Sprague-Dawley rats in a test similar to acute standard method (i.e. OECD TG 402, non GLP). The test substance was administered to the skin for 24 hours under occlusive conditions at doses of 740, 1110, 1670 and 2500 mg/kg bw to 4 groups of 10 animals (5animals/sex).

Clinical signs (dyspnea, agitation, apathy, atonia (with tremor and convulsions), hypothermia, cachexia) were observed starting from the mid-dose group. Within the 14 days observation period no animals died after single application of 740 mg/kg bw. 1/10, 8/10 and 9/10 animals died using 1110, 1670 and 2500 mg/kg bw respectively. Deaths occurred within 1 to 5 days post application. Having this results a median lethal dose (LD50) of 1500 mg/kg bw was identified for acute dermal toxicity.

Based on these results, the test substance is considered as harmful in contact with skin.

Testing for acute toxicity after exposure through inhalation a limit test equivalent to the one set out in OECD TG 403 was performed . None of the six animals (male and female rats) exposed for 8 hours to saturated vapour containing 3.1 mg test item per l air (nominal concentration) died during the 14 day observation period. Having these results one could state that under the conditions tested the median lethal dose of the test item is > 3.1 mg/l.


Justification for selection of acute toxicity – oral endpoint
reliable study (category 2), revealing the lowest dose descriptor of all available studies

Justification for selection of acute toxicity – inhalation endpoint
Only on study available.

Justification for selection of acute toxicity – dermal endpoint
Only one study available.

Justification for classification or non-classification

Considering the data described above classification for acute oral and acute dermal toxicity into category 4 (H302 and H312) is necessary according to Regulation (EC) No 1272/2008. In a test for acute toxicity after inhalation exposure no mortality was observed up to the highest feasible concentration tested therefore no calssification has to be considered according to Regulation (EC) No 1272/2008.

Corresponding to the data observed the submission substance has to be classified as harmful in contact with skin and if swallowed (risk phrases R21/22) according to the criteria set in Council Directive 67/548/EEC. Based on Council Directive 67/548/EEC no classification for acute toxicity after inhalation exposure is necessary.