Registration Dossier

Administrative data

Description of key information

ACUTE ORAL
LD50 > 2000 mg/kg bw (rat); OECD 423, EU Method B.1 tris, EPA OPPTS 870.1100 and JMAFF 12 Nousan, Notification No. 8147
ACUTE DERMAL
LD50 > 2000 mg/kg bw (rat); OECD 402, EU Method B.3, EPA OPPTS 870.1200 and JMAFF 12 Nousan, Notification No. 8147

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:
08 August 2014 to 02 September 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: JMAFF, 12 Nousan, Notification No 8147
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Strain: Wistar strain Crl:WI (Han) (outbred, SPF-Quality)
- Age at study initiation: Approximately 8 weeks old
- Weight at study initiation: 151 to 168 g. Body weight variation did not exceed ± 20 % of the sex mean.
- Fasting period before study: Animals were deprived of food overnight prior to dosing and until 3 to 4 hours after administration of the test material. Water was available ad libitum.
- Housing: Group housing of 3 animals per cage in labelled cages (height 18 cm) containing sterilised sawdust as bedding material and paper as cage-enrichment.
- Diet: Free access to pelleted rodent diet
- Water: Free access to tap water
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 18 to 24 °C
- Humidity: 40 to 70 % (relative)
- Air changes: at least 10 air changes/hour
- Photoperiod: 12-hour light/12-hour dark cycle
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 2.073 mL/kg bw. The dose volume was calculated as dose level (g/kg) / density (g/mL).

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Starting dose = 2000 mg/kg bw. Substance initially assumed to be of low toxicity based on test on amixture containing the substance.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
6 females; the test material was administered to two subsequent groups of three female rats
Control animals:
no
Details on study design:
- Duration of observation period following administration: 15 days
- Frequency of observations and weighing: Animals were observed for mortality and viability twice daily. Body weights were recorded on Day 1 (pre-administration of the test material) and on Days 8 and 15. At periodic intervals on the day of dosing (Day 1) and once daily thereafter until Day 15, animals were observed for clinical signs. The signs were graded according to fixed scales and the time of onset, degree and duration were recorded.
- Necropsy of survivors performed: Yes. At the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy.
- Other examinations performed: Descriptions of all internal macroscopic abnormalities were recorded.
Statistics:
No statistical analysis was performed as the method used is not intended to allow the calculation of a precise LD50 value.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
Hunched posture and piloerection were noted for all animals between Days 1 and 8. Lethargy and/or lean appearance were noted for one animal on Day 5 and/or 6.
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 revealed adhesions between the stomach and diaphragm in all animals.

Table 1: Body Weight Data (g)

Dose Group

Animal Number

Day 1

Day 8

Day 15

First group dosed with 2000 mg/kg bw

1

152

162

176

2

159

179

195

3

168

176

207

Mean (SD)

160 (8)

172 (9)

193 (16)

Second group dosed with 2000 mg/kg bw

4

152

165

197

5

151

170

203

6

155

165

198

Mean (SD)

153 (2)

167 (3)

199 (3)

SD = Standard deviation

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study the LD50 value was experimentally establised to exceed 2000 mg/kg bw and in accordance with OECD Guideline 423, the LD50 cut-off value was considered to be 5000 mg/kg
Executive summary:

The potential of the test material to cause acute oral toxicity in the rat was investigated in accordance with the standardised guidelines OECD 423, EU Method B.1 tris, EPA OPPTS 870.1100 and JMAFF 12 Nousan, Notification No. 8147 under GLP conditions.

The test material was administered by oral gavage to two subsequent groups of three fasted female Wistar rats at 2000 mg/kg body weight. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was conducted after terminal sacrifice (Day 15).

No mortality occurred. Hunched posture and piloerection were noted for all animals between Days 1 and 8. Lethargy and/or lean appearance were noted for one animal on Day 5 and/or 6. 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. Macroscopic post mortem examination revealed adhesions between the stomach and diaphragm in all animals.

Under the conditions of this study the LD50 value was established to exceed 2000 mg/kg bw and according to OECD Guideline 423 the LD50 cut-off value was considered to be 5000 mg/kg.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Both studies were conducted in accordance with standardised guidelines under GLP conditions. The quality of the database is therefore considered to be high.

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Both studies were conducted in accordance with standardised guidelines under GLP conditions. The quality of the database is therefore considered to be high.

Additional information

Acute Oral Toxicity

In the key study, the potential of the test material to cause acute oral toxicity in the rat was investigated in accordance with the standardised guidelines OECD 423, EU Method B.1 tris, EPA OPPTS 870.1100 and JMAFF 12 Nousan, Notification No. 8147 under GLP conditions. The study was awarded a reliability score of 1 in accordance with the principles for assessing data quality set forth by Klimisch et al. (1997).

The test material was administered by oral gavage to two subsequent groups of three fasted female Wistar rats at 2000 mg/kg body weight. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was conducted after terminal sacrifice (Day 15).

No mortality occurred. Hunched posture and piloerection were noted for all animals between Days 1 and 8. Lethargy and/or lean appearance were noted for one animal on Day 5 and/or 6. 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. Macroscopic post mortem examination revealed adhesions between the stomach and diaphragm in all animals.

Under the conditions of this study the LD50 value was established to exceed 2000 mg/kg bw and according to OECD Guideline 423 the LD50 cut-off value was considered to be 5000 mg/kg.

In the supporting GLP study conducted according to the standardised guidelines OECD 401 and EU Method B.1, the test material (approximately 65 % registered substance, 35 % xylene), at 2 g/kg bw in 10 mL maize oil/kg bw, was given by stomach tube to groups of 5 rats/sex, and the rats were observed for 14 days. The study was awarded a reliability score of 2 in accordance with the principles for assessing data quality set forth by Klimisch et al. (1997).

One female rat died on day 3 of the observation period, after displaying prone position, pallor, gasping, piloerection, ungroomed fur, pigmented staining of the snout and closed eyes. The cause of death could not be conclusively established (due to cannibalism), but the other females showed no signs of toxicity. There were no deaths, but reduced activity, piloerection and hunched posture were seen in five males on day 1, and staggering gait and salivation were seen in 2/5 and 1/5 males, respectively. However, there was complete recovery in all males by day 2.

The surviving animals, with the exception of one male, generally achieved anticipated body weight gains. The single case of reduced body weight gain could not be unequivocally attributed to treatment in the absence of a similar effect in other animals. Upon necropsy, there were no abnormalities in the major organs of the surviving animals.

Based on these results, the acute oral LD50 value for the test material in the rat is considered to be greater than 2 g/kg bw.

Acute Inhalation Toxicity

In accordance with Section 8.5.2. of Column 2 of Annex VIII of the REACH Regulation, the registrant proposes to waive the acute toxicity testing by the inhalation route on the grounds that exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and the possibility of exposure to aerosols, particles or droplets of an inhalable size.

Acute Dermal Toxicity

In the key study, the potential of the test material to cause acute dermal toxicity in the rat was investigated in accordance with the standardised guidelines OECD 402, EU Method B.3, EPA OPPTS 870.1200 and JMAFF 12 Nousan, Notification No 8147 under GLP conditions. The study was awarded a reliability score of 1 in accordance with the principles for assessing data quality set forth by Klimisch et al. (1997).

The test material was administered to five Wistar rats of each sex by a single dermal application at 2000 mg/kg body weight for 24 hours. After the exposure period, dressings were removed and the skin cleaned of residual test material using tap water. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15)

No mortality occurred. Chromodacryorrhoea (eye or snout) was noted in the majority of animals. One animal (No. 1) showed chromodacryorrhoea of the left eye before dosing. General erythema, erythema maculate, fissures, scales and/or scabs were seen in the treated skin-areas, left hindleg or left flank of the animals during the observation period.

All animals showed bodyweight loss or reduced bodyweight gain (two females) between Days 1 and 8. The changes noted in body weight in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity. No abnormalities were found at macroscopic post mortem examination of the animals.

Under the conditions of this study the LD50 value was found to exceed 2000 mg/kg bw.

In the supporting GLP study conducted according to the standardised guidelines OECD 402 and to EU Method B.3, the test material (approximately 65 % registered substance and 35 % xylene) was applied, occluded for 24 h, to the skin of rats (5/sex) and the animals observed for 14 days. The study was awarded a reliability score of 2 in accordance with the principles for assessing data quality set forth by Klimisch et al. (1997).

Apart from signs of significant local irritation (including erythema, oedema, exfoliation, eschar formation, sloughing, sensitivity and scarring) there were no other indications of toxicity; no deaths or systemic effects were seen, there were no abnormalities in the major organs and body weight gains were as expected. Based on these results, the acute dermal LD50 for the test material is greater than 2 g/kg bw in the rat.


Justification for selection of acute toxicity – oral endpoint
Two studies are available to address this endpoint. The key study was selected on the basis that the material tested was more representative of the substance being registered, with a purity of 97 %. The supporting study was conducted using material that consisted of 65 % of the registered substance and 35 % xylene.

Justification for selection of acute toxicity – dermal endpoint
Two studies are available to address this endpoint. The key study was selected on the basis that the material tested was more representative of the substance being registered, with a purity of 97 %. The supporting study was conducted using material that consisted of 65 % registered substance and 35 % xylene.

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No. 1272/2008, the substance does not require classification with respect to acute toxicity via the oral and dermal routes.