Registration Dossier

Diss Factsheets

Administrative data

Description of key information

The acute oral LD50 to male and female rats was > 2000 mg/kg bw, It can be concluded that product Shale oils, light is not toxic after single oral administration and belongs to class 5 (unclassified) according to GHS.
The median lethal dose of Shale oil after single dermal administration to rats of both sexes, observed over a period of 14 days is: LD50 (rat): greater than 2000 mg/kg body weight (this result is read-across from Shale oil middle fraction).
The median lethal dose of Generator oil after 2 hours exposure to white mice, observed over a period of 14 days was 19 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:
5th March - 14th April 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted 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
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Accredited Breeding Velaz, Prague, Czech Republic
- Age at study initiation: NDA
- Weight at study initiation: Males 175 - 190 g, females 150 - 155 g
- Fasting period before study: overnight prior to study
- Housing: Animals were housed 3 to a cage with wooden grate bedding and in conformity with animal welfare legislation.
- Diet (e.g. ad libitum): A standard certified laboratory diet was served. The diet is routinely analysed by the manufacturer for nutritional components and environmental contaminants.
- Water (e.g. ad libitum): ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2 ºC
- Humidity (%): 55 ± 10 %
- Air changes (per hr): 10 per hour
- Photoperiod (hrs dark / hrs light): 12 hours light / 12 hours dark

IN-LIFE DATES: From: 18th March 2010 To: 7th April 2010
Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200 mg/ml
- Amount of vehicle (if gavage): 10 ml/kg bw
- Justification for choice of vehicle: NDA
- Lot/batch no. (if required): L 812160
- Purity: NDA

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg bw

DOSAGE PREPARATION (if unusual): Test article was mixed with vehicle shortly before administration. Required amount of test article was weighed out and mixed with precise amount of vehicle shortly before application.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
6 (3 male/3 female) animals per dose
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 7 and 14 days after application
- Necropsy of survivors performed: yes
- Other examinations performed:
- Clinical signs:
Animals were observed individually after dosing at least once during the first 30 minutes, periodically during the first 24 hours, with special attention given during the first 4 hours and daily thereafter, for a total of 14 days. Duration of observation was considered sufficient. All observations are systemically recorded. Clinical signs and conditions associated with pain, suffering and impending death, are described in detail in a separate OECD Guidance Document. Observations included changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems, and somatomotor qactivity and behaviour pattern. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy sleep and coma.

- Body weight:
Individual weights of animals were determined shortly before the test article was administered and weekly thereafter. Weights of animals were recorded at the end of the study. Weight changes after first and second weeks after application were calculated and recorded.

- Pathology:
After finishing of observation period, the animals were sacrificed by exsanguinations in ether narcoses. The necropsies were made by standard operation procedures. All tested animals were subjected to a full, detailed gross necropsy, which included carful examination of external surface of the body, all orifices, and cranial, thoracic and abdominal cavities and their contents. All gross pathological changes will be recorded for each animal.
Statistics:
NDA
Preliminary study:
NDA
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
None observed
Clinical signs:
None observed
Body weight:
Smaller than expected body weight gains in males and a 5g body weight loss in one female were recorded 1 week after administration. Anticipated body weights were observed during the second week after adminstration.
Gross pathology:
One male had an enlarged spleen with an uneven surface. One female was found with an uneven surface on the spleen. No other effect were found.
Other findings:
NDA.

Table 1: Body weight changes

Cage

No.

Sex

Animal

No.

B.W.

initial

[g]

Applied

volume

[ml]

Response

 

    After 1 week

    After 2 weeks

B.W.

[g]

B.W. change

[g]

B.W.

[g]

B.W. change

[g]

3

Females

1

150

1.50

0

150

0

170

20

2

155

1.55

0

150

-5

155

5

3

155

1.55

0

170

15

185

15

4

Males

4

175

1.75

0

180

5

225

45

5

180

1.80

0

200

20

250

50

6

190

1.90

0

210

20

255

45

Response: 0 - surviving

X - death

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The LD50 to male and female rats was > 2000 mg/kg bw, It can be concluded that product Shale oils, light is not toxic after single oral administration and is therefore not classified according to the EU CLP criteria.
Executive summary:

Shale oils, light fraction was dosed orally via gavage to 3 female (150 - 155 g) and 3 male (175 - 190 g) wistar rats at limit dose of 2000 mg/kg, in accordance with OECD 423 and performed to GLP. Neither death nor visible symptoms of toxicity during 14 day observation period were seen. It can be concluded that product Shale oils, light is not toxic after single oral administration and is therefore not classified according to the EU CLP criteria.

Endpoint conclusion
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
not reported
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study conducted in accordance with generally accepted scientific principles, possibly with incomplete reporting or methodological deficiencies, which do not affect the quality of the relevant results. Study read across from generator oil.
Qualifier:
no guideline followed
Principles of method if other than guideline:
Acute inhalation experiments using mice in 2 hour exposures.
GLP compliance:
no
Limit test:
no
Species:
mouse
Strain:
other: white mice
Sex:
not specified
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
not specified
Vehicle:
not specified
Analytical verification of test atmosphere concentrations:
not specified
Duration of exposure:
2 h
Concentrations:
See Any other information on materials and methods
No. of animals per sex per dose:
110 animals used in the tests
Control animals:
not specified
Dose descriptor:
LC50
Effect level:
19 mg/L air
Based on:
test mat.
Exp. duration:
2 h
Remarks on result:
other: ± 1.4 mg/L

The influence of volatile compounds of generator oil the activity of the movement and irritation of mucosa was observed already at the concentration of 0.18 - 0.25 mg/l. Increasing the concentration of volatile compounds in the chamber up to 5.9 ml/l the mice looked tired (sluggish) during the test and 30 min after the test. In some cases during the second hour of the test dyspnoea was observed. At higher concentrations (10-12 ml/l) in addition to tiredness and dyspnoea, during the second hour of the test the redness of nose, ears and paws was observed.

Increasing the concentration of the volatile compounds up to 29 ml/l and higher during the second hour of the test additionally to lower movement activity, interruptions in movement coordination and dyspnoea in mice ballisms (mostly of the head and paws) were observed; after 10 , 15 and 20 minutes they lost the ability to stand on their feet (took side position). Afterwards the intoxicative appearance and strengthening of spasms was observed. Before death the convulsions gradually stopped. Animals died when breathing progressively weakened. In few cases death came 2-3 days after the test.

In autopsy of white mice, stagnation phenomenon in brain and lungs were observed. Quite often macroscopic signs of degenerative changes in liver were observed.

First signs of intoxication with volatile compounds of generator oil in white mice were observed at concentrations of 0,25 mg/l. The main parameters of toxicity to white mice were as follows: LC0 = 13 mg/l, LC100 = 30 mg/l, LC50 = 19 ± 1.4 mg/l.

As described before we can see that intoxication with volatile compound of generator oil are characterized with the same symptoms as of intoxication with other substances. These kind of symptoms as change in the character of breathing, disorder of movement coordination, cramps, lying on the side etc. are also signs of intoxication with organic solvents; redness of nose, ears and paws are observed in case of intoxication with toluene; head and paw cramps indicate impact of phenols etc.

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The LC50 of generator oil to white mice was 19 ± 1.4 mg/L.
Executive summary:

White mice were exposed to generator oil in an aerosol inhalation experiment for 2 hours.

The LC50 of generator oil to white mice was 19 ± 1.4 mg/L.

Endpoint conclusion
Dose descriptor:
LC50
Value:
19 000 mg/m³

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
8th April - 12th August 2005
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted 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. Study conducted on middle weight fraction of shale oils.
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. QA statement)
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: RCC Ltd, Laboratory Animal Services CH-4414 Füllinsdorf / Switzerland
- Age at study initiation: Males: 8 weeks. Female: 12 weeks
- Weight at study initiation: NDA
- Fasting period before study: NDA
- Housing: During acclimatization in groups of five per sex in Makrolon type-4 cages with standard softwood bedding. Individually in Makrolon type-3 cages with standard softwood bedding during treatment and observation.
- Diet (e.g. ad libitum): Pelleted standard provided ad libitum
- Water (e.g. ad libitum): Community tap water from Füllinsdorf ad libitum.
- Acclimation period: 8th - 12th April 2005

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 5
- Humidity: 30 - 70 %
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12 hours light / 12 hours dark with music played during the daytime period

IN-LIFE DATES: From: 8th April 2005 To: 27th April 2005
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: NDA
- % coverage: 10 %
- Type of wrap if used: The dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): skin flushed with luke warm water
- Time after start of exposure:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2.04 mL/kg bw
- Concentration (if solution): N/A
- Constant volume or concentration used: yes
- For solids, paste formed: N/A

VEHICLE
N/A
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 per sex per dose
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: on days 1, 8 and 15
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, local signs
Statistics:
No statistical analysis used.
Preliminary study:
not applicable.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
None
Clinical signs:
No systemic signs of toxicity were observed during the course of the study.
Due to brown discoloration of the application site in all animals, local symptoms could not be assessed on test day 2.
Slight general erythema was observed in six animals at the reading on test day 3 and persisted up to test day 5, 6, 10 or 15 and was noted additionally from test day 7 to 8 in one of these animals. Moderate general erythema was found in four animals on test day 3 and was still noted in two animals at the reading on test day 4 or 5. One female showed this symptom on test day 6 and 7. In these last four animals, slight general erythema was found from the reading on test day 4, 5, 6 or 8 until test day 6, 8, 10 or 15.
Slight formation of fissures was seen in eight animals at the reading on test day 3 or 4 and persisted up to the observation on test day 5 in one animal or test day 6 in the remaining animals.
Slight formation of crusts was observed in eight animals from the reading on test day 3, 4, or 6 up to the observation on test day 6, 7, 8 or 12 and returned in four of these animals from test day 8, 9, 11 or 13 to test day 12 or the end of the observation period. In three of these animals moderated crust formation was seen at the reading on test day 7 and persisted up to the observation on test day 8 in one animal.
Moderate crust formation was observed in two animals on the reading from test day 3 to test day 5 or on test day 7 only in one animal. Slight crust formation occurred in these animals from test day 6 to 7 or test day 13 to 15, respectively.
Slight scaling was observed in all animals from test day 8, 9, 12 or 13 to the end of the observation period.
(see attached tables)
Body weight:
The body weight of the animals was within the range commonly recorded for this strain and age.
Gross pathology:
No macroscopic findings were observed at necropsy.
Other findings:
NDA
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The median lethal dose of Shale oil after single dermal administration to rats of both sexes, observed over a period of 14 days is:
LD50 (rat): greater than 2000 mg/kg body weight.
Executive summary:

Five male and five female HanRcc:WIST (SPF) rats were treated with Shale oil at 2000 mg/kg by dermal application in accordance with OECD 402 and to GLP standard. The test item was applied undiluted as delivered from the sponsor at a volume dosage of 2.04 mL/kg. The application period was 24 hours.

No mortality, clinical signs or abnormal body weight changes were observed. Mild to moderate skin irritation effects were observed in all animals.

The median lethal dose of Shale oil after single dermal administration to rats of both sexes, observed over a period of 14 days is:

LD50 (rat): greater than 2000 mg/kg body weight.

Based on the rationale for read-across, it is considered acceptable to use this study to address the same endpoint for light fraction of shale oil.

Endpoint conclusion
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw

Additional information

Key values for Chemical Safety Assessment

Acute toxicity: oral

Effect level LD50 in mg/kg bw >2000

Acute toxicity: dermal

Effect level LD50 in mg/kg bw >2000

Acute toxicity: inhalation

Effect level LC50 in mg/m3= 19000

For both the light and middle fractions of shale oil, the acute oral LD50 was found to be >2000 mg/kg bw, and both were found not to induce gene mutations in a bacterial reverse mutation assay (AMES test) without and with metabolic activation. This demonstrates that the toxicological profile of the two fractions is similar. This, combined with analytical data which showed the two fractions to be compositionally similar, supports the validity of a read-across approach to address the acute dermal toxicity endpoint. Hence, the study conducted on the middle fraction of shale oil, is considered robust enough to be the key study for the light fraction.

Due to the physical and chemical similarity of generator oil to shale oils, it was considered acceptable to use a read across approach in order to address the acute inhalation toxicity endpoint.

Justification for classification or non-classification

Based upon the high LD50 values for oral, inhalation and dermal exposure, and the absence of other major significant effects, the available study data indicates that shale oils light fraction does not need to be classified for acute toxicity according to Regulation (EC) No 1272/2008. However, to take account of the potential variability in the composition of the UVCB starting material, and the potential variation in the composition of the UVCB substance in scope of this registration, the worst case classification of H301 (Toxic if swallowed), H311 (Toxic in contact with skin), H332 (Harmful if inhaled) and H304 (May be fatal if swallowed and enters airways) is applied.


For further information, please refer to the document 'Shale oils RAAF Report' as included in section 13.