Registration Dossier

Administrative data

Description of key information

Studies on repeated dose toxicity have therefore been waived in consideration of the provisions provided in Annex XI of the regulation, Section 1 (testing does not appear scientifically necessary) on the basis of section 1.1 “use of existing data” and section 1.2 “weight of evidence” for the following reasons: 


1. An assessment of the hazard classifications of all constituents within shale oil did not indicate any concern regarding repeated dose toxicity.


2. The PAH component of shale oil is considered to drive the long term toxicological hazards for the registered substance. Since benzo[a]pyrene was the only identified PAH, and bearing in mind its stringent hazard classification, as part of a conservative approach, this substance has been used as the surrogate PAH substance within shale oil. Available repeated doe toxicity data on benzo[a]pyrene are therefore included within the substance dataset and are considered to provide conservative assessment of long term toxicological hazard of the registered substance.


3. The OEL for benzo[a]pyrene has been used to derive the relevant DNELs for the registered substance and has been shown to provide more conservative values than utilising the Virtually Safe Dose as determined by the Dutch authorities following long term testing with benzo[a]pyrene.


4. Upon consideration of the CMR (category 1B) classification, strict operating controls and rigorous risk management measures are implemented on site to ensure safe use, to mitigate CMR risk. These measures are considered protective for any other potential toxicological risk, including repeated dose toxicity.


 


In summary, the risk assessment is performed based on the most toxicologically relevant component, benzo(a)pyrene. Classification and DNELs can be derived based on calculation from the OEL recognised at European level. Even at 1% , the presence of PAHs are still the only component that need to be taken into consideration in the derivation of DNELs, because comparison of existing reference doses and exposure limits show that for PAHs are 10,000 lower than next lowest reference dose i.e. extrapolating the OEL for each oil fraction from the PAH content still results in the lowers OEL, than if were done based on the content of the remaining materials.


 


Please refer to the document ‘Consideration of the long term toxicity of Distillates (shale oil)’, as included in section 13, for further information.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
short-term repeated dose toxicity: oral
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Justification for type of information:
JUSTIFICATION FOR DATA WAIVING
The repeated dose toxicity data requirements of REACH are waived on the basis of the provisions provided in Annex XI of the regulation, Section 1 (testing does not appear scientifically necessary) on the basis of section 1.1 “use of existing data” and section 1.2 “weight of evidence”. The following further outline the relevant justifications:

1. Consideration of the substance compositions and the hazard classifications of the constituent substances
The hazard classifications of the constituent substances of the shale oil fractions were assessed and none indicated the need to classify for specific target organ toxicity following repeated exposure (please refer to the document ‘Shale oils RAAF Report’, as included in section 13, for further information).

2. Consideration of the presence of PAH in each of the registered substances and the view that PAHs are the drivers for the long term toxicity hazards of the registered substances
Analytical information on the registered substances shows that PAHs are present at 1% in the registered substance. Benzo[a]pyrene was the only PAH identified (heavy fraction analysis). Bearing in mind the accepted opinion that PAHs can be grouped in view of the similarities in their toxicity profiles, and in view of the particularly stringent harmonised classification of benzo[a]pyrene, benzo[a]pyrene data are used as a surrogate for the PAH content of the registered substances. Available repeated dose toxicity data on benzo[a]pyrene, as generated by the Dutch authorities (RIVM, 2001), is therefore included in the dataset of the registered substance and is considered to provide conservative assessment of long term toxicological hazard of the registered substancs. The data are included to support the repeated dose toxicity waivers and have been added as part of the general weight of evidence approach. The available study data on benzo[a]pyrene are considered to be equivalent to data generated by corresponding test methods referred to in Article 13(3) since they were conducted on behalf of the Ducth authorities and are:
1) adequate for the purpose of classification and labelling and/or risk assessment;
2) adequate and reliable in terms of coverage of the key parameters foreseen to be investigated in the corresponding test methods referred to in Article 13(3);
3) the exposure durations were comparable to or longer than the corresponding test methods referred to in Article 13(3) if exposure duration is a relevant parameter; and
4) adequate and reliable documentation of the studies are provided.

3. Consideration of the OEL that has been utilised in generation of conservative DNELs
OEL (occupational exposure limit) values for benzo[a]pyrene have been established by the European Agency for Safety and Health. In this report the lowest OEL value for 8 hours exposure is 0.002 mg/m³. It is considered this value can be viewed as adequately protective to the registered substance since benzo[a]pyrene can be considereda conservative surrogate for the PAH component, which itself can be considered to be the most hazardous component.

Additional repeated dose toxicity data are therefore not considered to be required in order to generate a DNEL. Furthermore, this approach is more conservative than the Virtually Safe Dose (VSD) of 5 ng benzo[a]pyrene/kg bw/day based on the results from the RIVM 2-year rat study (and are comparable to VSD based on the 2-year mice study).
In summary, the risk assessment is performed based on the most toxicologically relevant component, benzo(a)pyrene. Classification and DNELs can be derived based on calculation from the OEL recognised at European level. Even at 1%, the presence of PAHs are still the only component that need to be taken into consideration in the derivation of DNELs, because comparison of existing reference doses and exposure limits show that for PAHs are 10000 lower than next lowest reference dose i.e. extrapolating the OEL for each oil fraction from the PAH content still results in the lowest OEL, than if were done based on the content of the remaining materials.

4. Consideration of CMR (category 1B) classification and the stringent operational controls and risk management measures implemented during use to mitigate risk
In view of the CMR category 1B classifications that have been assigned to the registered substance, together with conservative approach taken in classifying for other REACH endpoints, it is not considered to be an expedient use of animals to commission repeated dose toxicity studies on the registered substance. Comprehensive operational controls and risk management measures are implemented at sites using the substances, to ensure risk mitigation of the CMR (category 1B) hazard. The measures that are implemented to ensure safe use of a CMR category 1B substance are considered protective for any potential repeated dose toxicity hazard. Any information which might be generated from repeated dose toxicity studies would not provide data that would improve the safety of workers who might be exposed to the substances.
The exposure scenarios, as included in chapters 9 and 10 of the substance CSRs, cover all relevant exposures throughout the lifecycle of the substance and demonstrate that the substance is not incorporated into an article. In all cases, comprehensive OCs/RMMs are enforced to the extent that risk to humans and to the environment is mitigated.

In addition to the column 2 and Annex XI justifications outlined above. The waivers are further justified on exposure considerations and animal welfare grounds.

Exposure considerations
The level of potential exposure was modelled (using ART), taking account of the relevant operational controls (OCs) and risk management measures (RMMs) that are applied. In all cases, comprehensive OCs/RMMs are enforced to the extent that there is minimal opportunity for exposure during use of the substance to humans and to the environment.
All workers receive training in the handling of the substances and all workers are required to wear an appropriate level of personal protective equipment, as a secondary means of protection to the primary technical means of control. All personal protective equipment conforms to European Standards and are selected in consideration of the exposure period, the expected level of exposure and are made of the appropriate material/ have the appropriate Assigned Protection Factor (AF).
As part of a qualitative exposure assessment of human health, which was conducted as a means of providing an additional level of assurance on the lack of risk, the potential (worst case estimates for) exposures were compared to the relevant OELs. In all cases, the exposure values predicted for ‘shale oil’ are considerably less than the OELs that are used for monitoring purposes at the facilities using shale oil and, as such, the use of the substances are not considered to pose a risk to the health of workers.
There are no consumer uses of the substances.
Overall, no risk to human health is anticipated in view of the operational controls and risk management measures that are implemented. Furthermore, chemical monitoring programs at the use sites confirms that workers are not exposed to any of the substances at levels above the OELs.

Animal welfare considerations
The protection and welfare of animals is an area covered by a wide range of EU legislation. Replacement, Reduction and Refinement (the “Three Rs”) is the key strategy of a systematic framework aimed at achieving the goal of humane experimental techniques. In order to improve the welfare of animals still needed for scientific purposes, the aim is to firmly anchor the principle of the “Three Rs” in EU legislation. In this context, Regulation (EC) No 1907/2006, Article 25 (1), clearly states that animal testing is to be avoided and only allowed as a last resort. The use of existing data as part of a weight of evidence approach has indicated that the use of benzo[a]pyrene data can be considered appropriate and protective for the mitigation of risk following long-term and repeated exposure to the registered substances. Further testing with the registered substances is therefore not considered to be an expedient, or responsible, use of animals.

Please refer to "Justification for type of information" and also to the attached document '1904651.UK0 - 7899 Consideration of the long term toxicity of Distillates (shale oil)' for further information.
Endpoint:
chronic toxicity: oral
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Justification for type of information:
JUSTIFICATION FOR DATA WAIVING
The repeated dose toxicity data requirements of REACH are waived on the basis of the provisions provided in Annex XI of the regulation, Section 1 (testing does not appear scientifically necessary) on the basis of section 1.1 “use of existing data” and section 1.2 “weight of evidence”. The following further outline the relevant justifications:

1. Consideration of the substance compositions and the hazard classifications of the constituent substances
The hazard classifications of the constituent substances of the shale oil fractions were assessed and none indicated the need to classify for specific target organ toxicity following repeated exposure (please refer to the document ‘Shale oils RAAF Report’, as included in section 13, for further information).

2. Consideration of the presence of PAH in each of the registered substances and the view that PAHs are the drivers for the long term toxicity hazards of the registered substances
Analytical information on the registered substances shows that PAHs are present at 1% in the registered substance. Benzo[a]pyrene was the only PAH identified (heavy fraction analysis). Bearing in mind the accepted opinion that PAHs can be grouped in view of the similarities in their toxicity profiles, and in view of the particularly stringent harmonised classification of benzo[a]pyrene, benzo[a]pyrene data are used as a surrogate for the PAH content of the registered substances. Available repeated dose toxicity data on benzo[a]pyrene, as generated by the Dutch authorities (RIVM, 2001), is therefore included in the dataset of the registered substance and is considered to provide conservative assessment of long term toxicological hazard of the registered substancs. The data are included to support the repeated dose toxicity waivers and have been added as part of the general weight of evidence approach. The available study data on benzo[a]pyrene are considered to be equivalent to data generated by corresponding test methods referred to in Article 13(3) since they were conducted on behalf of the Ducth authorities and are:
1) adequate for the purpose of classification and labelling and/or risk assessment;
2) adequate and reliable in terms of coverage of the key parameters foreseen to be investigated in the corresponding test methods referred to in Article 13(3);
3) the exposure durations were comparable to or longer than the corresponding test methods referred to in Article 13(3) if exposure duration is a relevant parameter; and
4) adequate and reliable documentation of the studies are provided.

3. Consideration of the OEL that has been utilised in generation of conservative DNELs
OEL (occupational exposure limit) values for benzo[a]pyrene have been established by the European Agency for Safety and Health. In this report the lowest OEL value for 8 hours exposure is 0.002 mg/m³. It is considered this value can be viewed as adequately protective to the registered substance since benzo[a]pyrene can be considereda conservative surrogate for the PAH component, which itself can be considered to be the most hazardous component.

Additional repeated dose toxicity data are therefore not considered to be required in order to generate a DNEL. Furthermore, this approach is more conservative than the Virtually Safe Dose (VSD) of 5 ng benzo[a]pyrene/kg bw/day based on the results from the RIVM 2-year rat study (and are comparable to VSD based on the 2-year mice study).
In summary, the risk assessment is performed based on the most toxicologically relevant component, benzo(a)pyrene. Classification and DNELs can be derived based on calculation from the OEL recognised at European level. Even at 1%, the presence of PAHs are still the only component that need to be taken into consideration in the derivation of DNELs, because comparison of existing reference doses and exposure limits show that for PAHs are 10000 lower than next lowest reference dose i.e. extrapolating the OEL for each oil fraction from the PAH content still results in the lowest OEL, than if were done based on the content of the remaining materials.

4. Consideration of CMR (category 1B) classification and the stringent operational controls and risk management measures implemented during use to mitigate risk
In view of the CMR category 1B classifications that have been assigned to the registered substance, together with conservative approach taken in classifying for other REACH endpoints, it is not considered to be an expedient use of animals to commission repeated dose toxicity studies on the registered substance. Comprehensive operational controls and risk management measures are implemented at sites using the substances, to ensure risk mitigation of the CMR (category 1B) hazard. The measures that are implemented to ensure safe use of a CMR category 1B substance are considered protective for any potential repeated dose toxicity hazard. Any information which might be generated from repeated dose toxicity studies would not provide data that would improve the safety of workers who might be exposed to the substances.
The exposure scenarios, as included in chapters 9 and 10 of the substance CSRs, cover all relevant exposures throughout the lifecycle of the substance and demonstrate that the substance is not incorporated into an article. In all cases, comprehensive OCs/RMMs are enforced to the extent that risk to humans and to the environment is mitigated.

In addition to the column 2 and Annex XI justifications outlined above. The waivers are further justified on exposure considerations and animal welfare grounds.

Exposure considerations
The level of potential exposure was modelled (using ART), taking account of the relevant operational controls (OCs) and risk management measures (RMMs) that are applied. In all cases, comprehensive OCs/RMMs are enforced to the extent that there is minimal opportunity for exposure during use of the substance to humans and to the environment.
All workers receive training in the handling of the substances and all workers are required to wear an appropriate level of personal protective equipment, as a secondary means of protection to the primary technical means of control. All personal protective equipment conforms to European Standards and are selected in consideration of the exposure period, the expected level of exposure and are made of the appropriate material/ have the appropriate Assigned Protection Factor (AF).
As part of a qualitative exposure assessment of human health, which was conducted as a means of providing an additional level of assurance on the lack of risk, the potential (worst case estimates for) exposures were compared to the relevant OELs. In all cases, the exposure values predicted for ‘shale oil’ are considerably less than the OELs that are used for monitoring purposes at the facilities using shale oil and, as such, the use of the substances are not considered to pose a risk to the health of workers.
There are no consumer uses of the substances.
Overall, no risk to human health is anticipated in view of the operational controls and risk management measures that are implemented. Furthermore, chemical monitoring programs at the use sites confirms that workers are not exposed to any of the substances at levels above the OELs.

Animal welfare considerations
The protection and welfare of animals is an area covered by a wide range of EU legislation. Replacement, Reduction and Refinement (the “Three Rs”) is the key strategy of a systematic framework aimed at achieving the goal of humane experimental techniques. In order to improve the welfare of animals still needed for scientific purposes, the aim is to firmly anchor the principle of the “Three Rs” in EU legislation. In this context, Regulation (EC) No 1907/2006, Article 25 (1), clearly states that animal testing is to be avoided and only allowed as a last resort. The use of existing data as part of a weight of evidence approach has indicated that the use of benzo[a]pyrene data can be considered appropriate and protective for the mitigation of risk following long-term and repeated exposure to the registered substances. Further testing with the registered substances is therefore not considered to be an expedient, or responsible, use of animals.

Please refer to "Justification for type of information" and also to the attached document '1904651.UK0 - 7899 Consideration of the long term toxicity of Distillates (shale oil)' for further information.
Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
not reported
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail
Remarks:
Method and results were sufficiently described, similar to OECD-guideline 407. The main deviations from the current test guidelines include a lack of neurobehavioral investigations and a limited number of organs weighed and examined at the terminal investigations.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
no guideline followed
Principles of method if other than guideline:
Groups of 10 animals per sex, per dose, received doses of test material, by oral gavage, 5 days a week. The animals were observed daily and their behaviour and clinical signs were examined. Body weights, food and water consumption was measured weekly. During the sixth week of exposure the animals were killed by exsanguination under ether anesthesia and blood, urine and tissues were taken for investigations on standard toxicological parameters. All animals were subjected to macroscopical examination, organ weights were recorded, and tissue samples were processed for further histopathological investigations. In addition, liver enzyme induction was monitored by EROD (ethoxyresurfin-O-deethylase) activity in plasma.
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
The dose-range finding study was performed with B[a]P from Janssen Chimica (Beerse, Belgium), which had a purity of 97.7 %.
Species:
rat
Strain:
Wistar
Remarks:
SPF Riv:TOX
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: the animals were bred at and derived from the Animal Facility at the Institute
- Age at study initiation: 4-5 weeks at acclimation initiation, 6 weeks of age at the initiation of dosing
- Housing: animals were housed in macrolon cages with a wire floor, two per cage
- Diet: ad libitum (SSP-Tox, Hope Farms BV, Woerden). From the start of treatment the applied food contained a reduced amount of soy oil to compensate for the soy oil used to administer the test material.
- Water: ad libitum (tap water - public drinkingwater, WMN, Utrecht)
- Acclimation period: 10 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25 °C
- Humidity (%): 40 - 72 %
Route of administration:
oral: gavage
Details on route of administration:
The length of the gavage needle used assured exposure of (at least the caudal part of) the oesophagus (a known target-site for carcinogenesis). Application was always in the morning for all groups and took about 0.5 -1 hour.
Vehicle:
soya oil
Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS: The test material was dissolved in soy oil in volumes of 0.4 and 0.5 mL per female and male rat, respectively. The test material was dissolvable in soy oil up to ≈ 30 g/L.
Fresh solutions were prepared weekly.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The stability of B[a]P under conditions of dissolution and storage (once dissolved) was verified. Due to several corrections (e.g. purity) actually achieved dose levels were a few percent below those targeted. No detailed analytical method was reported.
Duration of treatment / exposure:
5 weeks
Frequency of treatment:
5 days a week
Dose / conc.:
1.5 mg/kg bw/day (nominal)
Dose / conc.:
5 mg/kg bw/day (nominal)
Dose / conc.:
15 mg/kg bw/day (nominal)
Dose / conc.:
50 mg/kg bw/day (nominal)
No. of animals per sex per dose:
Groups of 10 animals (per dose, per sex)
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: animals were ad random assigned to the various dose groups by standardised procedures
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations included assessment of behaviour and clinical symptoms

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION: Yes
- Time schedule for examinations: weekly

WATER CONSUMPTION: Yes
- Time schedule for examinations: weekly

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: during the sixth week of exposure the animals were killed by exsanguination under ether anesthesia and blood, urine and tissues were taken for investigations on standard toxicological parameters
- Parameters examined included: Hb, Ht, RBC, MCV, MHC, MCHC, WBC, PLT

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: during the sixth week of exposure the animals were killed by exsanguination under ether anesthesia and blood, urine and tissues were taken for investigations on standard toxicological parameters
- Parameters examined included: GGT, ASAT, ALAT, LDH, and kreatinine

URINALYSIS: Yes
- Time schedule for collection of urine: during the sixth week of exposure the animals were killed by exsanguination under ether anesthesia and blood, urine and tissues were taken for investigations on standard toxicological parameters
- Parameters examined included: pH, protein, glucose, ketone, bilirubin, blood, nitrate, and urobilinogen

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes. All animals were subjected to macroscopical examination.
- Organ weights: liver, lung, thymus, spleen, kidneys, adrenals, and ovaries

HISTOPATHOLOGY: Yes. All animals of the highest dose groups and controls were histopathologically examined for their oesophagus, stomach, duodenum, liver, kidneys, spleen, thymus, lung and mammary gland (females only). In case of abnormalities the intermediate dose-groups were examined additionally.
Other examinations:
Liver enzyme induction was monitored by EROD (ethoxyresurfin-O-deethylase) activity in plasma
Clinical signs:
no effects observed
Description (incidence and severity):
No effects were observed during this 5-week treatment period, either on behaviour or upon handling.
Mortality:
no mortality observed
Description (incidence):
None of the animals died within the 5-week treatment period.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
There were no differences in body weight development between controls and test material-treated animals within the 5-week period.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
An increase in food consumption with treatment was observed for females at ≥ 5 mg/kg bw/day , whereas for males a decrease was found without a dose-response relationship. There are no explanations for the observed changes in food consumption.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
effects observed, non-treatment-related
Description (incidence and severity):
The treatment did not affect water-consumption by females, but it did reduce consumption by males (except for the highest dose), which was about 12% in excess over control value. There are no explanations for the observed changes in water consumption.
Ophthalmological findings:
not examined
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Apart from some non-statistical, small, dose-related decreases in Hb (both sexes), and RBC counts (males), treatment with the test material did not have any significant effects on investigated haematological parameters.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
Treatment with the test material did not have any significant effects on GGT, ASAT, and ALAT values. The observation of slight haemolysis within a substantial number of samples paralleled the variably increased LDH values. A small increase with treatment was observed on creatinine levels in males only.
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
Of the urinalysis parameters investigated, treatment with the test material only induced some increase in nitrite concentration in females.
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Treatment with the test material did not have any significant effects on organ-weights of lung, spleen, kidneys, adrenals, ovaria, and testis (data not shown). A reduction in thymus weight (in both sexes at 15 and/or 50 mg/kg bw/day) and an increase in liver weight (in both sexes at 50 mg/kg bw/day) was observed.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
No treatment-related effects could be discerned after the 5-week exposure period. Occasionally, some common background changes were obeserved; single cases of petechiae at the thymus, hydronephrosis, and a dilated uterus.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
FORESTOMACH: Small hyperplastic responses were observed in the basal layer of the epithelium, without any pattern and to a variable extent. This response was characterised by an increased number of cells with an oval rather than rounded nucleus and oriented perpendiculary to the basement membrane, or cells clearly showing hyperthrophy. Also, the regular occurrence of mitotic figures in the basal as well as suprabasal layer, vacuolization within and around the nucleus, nuclear polymorphy, atypia, and eosinophilic nucleoli reflected this mitotic response. Furthermore, the border with the lamina propria sometimes was irregular and not distinct due to nests of proliferating epithelial cells and/or influx of predominantly mononucleated inflammatory cells. A dose-response relationship was observed at and above 15 mg/kg bw/day.

LIVER: No clear treatment-related changes were observed in this organ. Therefore, intermediate dose-groups were not examined.

OESOPHAGUS: Inflammatory lesions, muscle regeneration, and atrophy were observed in a number of cases. As these are unusual, and show no relation with treatment with the test material, it is assumed that they are secondary to trauma by gavage application. Local mucosa appeared normal, probably due to a higher regerative capability.

THYMUS: An increased incidence of brown pigmentation of red pulp (hemosiderin) was observed in treated animals of both sexes. Because of the very small difference (as compared to controls), and the absence of any further support for this (e.g. haematological changes) intermediate groups were not examined.
Other effects:
effects observed, treatment-related
Description (incidence and severity):
Liver enzyme induction (EROD): treatment with the test material had a very pronounced effect on liver microsomal EROD activity. The induction level at a dose of 50 mg/kg bw appears to be the maximal achievable in male rats (under the present treatment conditions); a level which is about 36 times control values. Induction was already apparent (i.e. about 5 times increase) at the lowest tested dose of 1.5 mg/kg bw. In females the highest induction level appeared not to be reached at 50 mg/kg bw. In the absence of treatment with the test material (during the weekends) EROD activity appeared to rapidly decline to almost control values, indicating the dynamics of the induction mechanism.
Remarks on result:
not measured/tested
Critical effects observed:
not specified
Conclusions:
The study was conducted as a range-finding study for a subsequent 2-year carcinogenicity study. On the basis of findings from this study a top dose of 30 mg/kg bw/day was taken forward for the carcinogenicity study.
Executive summary:

A short term repeated dose toxicity study was conducted to help inform on dose selection for a subsequent carcinogenicity study. This range-finding study was conducted under GLP conditions.

During the study, groups of 10 animals per sex, per dose, received doses of test material, by oral gavage, 5 days a week. The animals were observed daily and their behaviour and clinical signs were examined. Body weights, food and water consumption was measured weekly. During the sixth week of exposure the animals were killed by exsanguination under ether anesthesia and blood, urine and tissues were taken for investigations on standard toxicological parameters. All animals were subjected to macroscopical examination, organ weights were recorded, and tissue samples were processed for further histopathological investigations. In addition, liver enzyme induction was monitored by EROD (ethoxyresurfin-O-deethylase) activity in plasma.

On the basis of findings from this study, particularly in relation to effects noted on basal cell hyperplasia in the forestomach as well as the effects upon liver- and thymus-weight, doses of 3, 10 and 30 mg/kg bw/day were proprosed for the carcinogenicity study. In order to verify the observed forestomach proliferation, morphometrical analysis of the forestomach was incorporated into a 90-day repeated dose toxicity stduy in rats using the BrdU-incorporation technique (see a robust study summary under section 7.5.1). This study essentially showed similar results.

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
not reported
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail
Remarks:
Method and results were sufficiently described, similar to OECD-guideline 408. The main deviations from the current test guidelines include a lack of thyroid hormone measurements and neurobehavioral investigations, as well as a limited number of organs weighed and examined at the terminal investigations.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
no guideline followed
Principles of method if other than guideline:
In conjunction with the 2-year carcinogenicity study, groups of 10 animals per sex, per dose, received doses of test material, by oral gavage, for 5 days a week over a period of 90 days. The animals were observed daily and their behaviour and clinical signs were examined. Body weights, food and water consumption was measured at least weekly. Following the end of the exposure period the animals were sacrificed by exsanguination from the abdominal aorta. Blood, urine and tissues were taken for investigations on standard toxicological parameters. All animals were subjected to macroscopic examination, organ weights were recorded, and tissue samples were processed for further histopathological investigation. The purpose of this 90-day kill was to reassure the appropriateness of the dose levels chosen for the chronic carcinogenicity study.
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
The dose-range finding study was performed with B[a]P from Janssen Chimica (Beerse, Belgium), which had a purity of 97.7 %.
Species:
rat
Strain:
Wistar
Remarks:
SPF Riv:TOX
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: the animals were bred at and derived from the Animal Facility at the Institute
- Age at study initiation: 4-5 weeks at acclimation initiation. Animals were 6 weeks of age at study initiation (following acclimation).
- Housing: animals were housed in macrolon cages with a wire floor, two per cage
- Diet: ad libitum (SSP-Tox, Hope Farms BV, Woerden). From the start of treatment the applied food contained a reduced amount of soy oil to compensate for the soy oil used to administer the test material (standard SSPTox food contains 4.55% soy oil on a weight basis).
- Water: ad libitum (tap water - public drinkingwater, WMN, Utrecht)
- Acclimation period: 10 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25 °C
- Humidity (%): 40 - 72 %
Route of administration:
oral: gavage
Details on route of administration:
The length of the gavage needle used assured exposure of the distal half of the oesophagus (a known target-site for carcinogenesis).
Application was always in the morning for all groups and took about 1.5 - 2.5 hours.
Vehicle:
soya oil
Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS:
The test material was dissolved in soy oil. At the start of treatment the test material was applied in 0.4 and 0.5 mL soy oil per female and male rat, respectively. Upon body weight gain and herewith amount of test material per rat these volumes were adjusted to 0.6 and 0.75 mL, respectively.
Solubility of test material in soy oil was ≈ 30 g/l.
Fresh solutions were prepared weekly.
Dosing was on the basis of extrapolated mean group weights (per dose, per sex).
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Test material solutions in use were regularly verified for their benzo[a]pyrene content.
Duration of treatment / exposure:
90-days
Frequency of treatment:
5 days per week
(Rats were not dosed at some national or christian celebration days)
Dose / conc.:
3 mg/kg bw/day (nominal)
Remarks:
Due to several corrections (e.g. purity) actually achieved dose levels were somewhat lower, i.e. 2.9 ± 0.3 mg/kg bw/day
Dose / conc.:
10 mg/kg bw/day (nominal)
Remarks:
Due to several corrections (e.g. purity) actually achieved dose levels were somewhat lower, i.e. 9.6 ± 1.0 mg/kg bw/day
Dose / conc.:
30 mg/kg bw/day (nominal)
Remarks:
Due to several corrections (e.g. purity) actually achieved dose levels were somewhat lower, i.e. 29 ± 3 mg/kg bw/day
No. of animals per sex per dose:
Groups of 10 animals (per dose, per sex)
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: animals were ad random assigned to the various dose groups by standardised procedures
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: animals were examined daily for behaviour and clinical symptoms and by palpation.

BODY WEIGHT: Yes
- Time schedule for examinations: every week (week 1 - 8), every 2 weeks thereafter

FOOD CONSUMPTION: Yes
- Time schedule for examinations: twice weekly

WATER CONSUMPTION: Yes
- Time schedule for examinations: twice weekly

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: On the day of sacrifice blood was taken under ether narcosis by orbita puncture.
- Parameters examined included: WBC, RBC, Hb, Ht, MCV, MCHC, PLT

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: On the day of sacrifice blood was taken under ether narcosis by orbita puncture.
- Parameters examined included: ALAT, ASAT, γ-GT, LD, creatinine

URINALYSIS: Yes
- Time schedule for collection of urine: On the day before sacrifice the animals received their last dose, and were allocated in metabolism cages for collection of urine.
- Parameters examined included: volume, creatinine and protein

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No
Sacrifice and pathology:
TERMINAL PROCEDURES: Prior (1h ± 15') to euthanasia the rats received an i.p. injection of 2 mL buffered saline containing 20 mg BrdU/mL. The animals were sacrificed by exsanguination from the abdominal aorta.

GROSS PATHOLOGY: Yes
The following organs were sampled: brain; pituitary; thyroid (attached to larynx); thymus; heart; lungs (after formalin infusion); liver (two lobes); spleen; pancreas; kidneys; adrenals; ovaries / testes; uterus / prostate with seminal vesicles; sciatic nerve; quadriceps muscle; gastro-intestinal tract (stomach opened and spread on paraffin wax disks; swiss rolls from oesophagus, duodenum; jejunum; ileum; caecum; colon and rectum); lymph nodes (axillary, mesenteric and mandibular); ventral abdominal skin (including subcutaneous mammary tissue) and femur.
All tissues were fixed in 4% neutral buffered formaldehyde.

ORGAN WEIGHTS: Yes
Liver, thymus and spleen were weighted.

HISTOPATHOLOGY: Yes
For initial histological examination the following tissues were designated as protocol organs: liver, stomach, esophagus, thymus, lung, spleen and mesenteric lymph node.
Other organs were archived separately for backup purposes (possible unexpected tumour yields) and possible other future research, in view of the unique character of the material.
Histological specimens of protocol organs were processed routinely (paraffin embedding and H&E staining of 5 μm sections). From the stomach, a putative target organ and selected for morphometric counting of BrdU labelling, two samples were taken perpendicular to the limiting ridge, containing both fore- and glandular stomach. Other samples were processed according to Standard Operation Procedures of the Laboratory of Pathology.
Initially only control and high dose groups animals were examined. Organs displaying changes that could be dose-related, were further analysed by "blind reading", and if a difference was apparent, also the intermediate dose groups were studied.
Morphometrical analysis of the forestomach was carried out for the prevalence of S-phase epithelial cells displaying BrdU incorporation after immunohistochemical staining. From each section of the forestomach five representative samples (if possible), each over the length of the monitor were counted, and the total per stomach was expressed in relation to the surface of the corresponding muscularis mucosae. The latter was done to compensate for variation in stretching of the stomach wall. Morphometry was performed using the IBAS 2000 image analysis system (Kontron Bildanalyse GmbH, Eching,
Germany).
Statistics:
(Morphometrical analysis of the forestomach) the data were analysed statistically by ANOVA for treatment and by Student-t test for group wise-comparison.
Clinical signs:
no effects observed
Description (incidence and severity):
No remarkable effects were observed during this 3-months treatment period, either on behaviour or upon handling.
Mortality:
no mortality observed
Description (incidence):
There were no differences in survival between controls and test material-treated animals within the 3-month period.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Only at the highest dose tested (30 mg/kg bw), the test material appeared to affect body weight gain in males only, i.e. from week 10 onwards. At the same dose level females appear to be less sensitive for this effect.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Treatment with the test material appeared to have no significant effect on food consumption.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
Treatment with the test material appeared to have no significant effect on water consumption.
Ophthalmological findings:
not examined
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Apart from some non-statistical, small, dose-related decreases in RBC count and Hb (both sexes), treatment with the test material did not have any significant effects on the investigated haematological parameters.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
No treatment-related group-differences, nor dose-response relationships could be discerned for ALAT, ASAT, LD, or creatinine. For γ-GT a small dose-related decrease in activity was found in males. The treatment-related small increase in creatinine levels observed in the range-finding study with males could not be reproduced under the present conditions.
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
Urinary volume was increased in males at the highest B[a]P-dose tested, though in the absence of a clear dose-response relationship. Urinary creatinine levels appeared increased at this treatment level in males and females. Both male and female rats showed a dose-related increase in urinary protein levels. In the absence of further investigations, underlying mechanisms are yet unresolved.
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Clear target-organs of toxicity were the liver and thymus i.e. increased liver- and decreased thymus weights; changes in weight of liver and thymus were generally confined to the highest dose level, except for the liver of males, in which also 10 mg/kg bw induced a substantial (15%) weight increase. Spleen weights were only just significantly increased (P < 0.05) at the top dose in both sexes.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
A consistent finding in most animals was a discolouration of the mandibular lymph nodes, and occasionally other regional lymph nodes (axillary). This discolouration varied from reddish to brown / grey, although in the table this discolouration was named brown / red. This phenomenon was attributed to minute local haemorrhages caused by manipulation (gavage), which was conveyed to the regional lymph node. Other changes occurred at a low incidence and were consistent with common background pathology.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
FORESTOMACH:
The epithelium of the forestomach of the 30 mg/kg group in particular demonstrated minimal changes that could be summarised as "basal cell disturbance". The diagnosis basal cell disturbance was based on the combination of various features that occurred either focal or diffuse. These features were: increase in number of basal cells, mitotic figures and remnants of necrotic cells; occasionally early nodule development; infiltration by inflammatory cells (mainly histiocytes); capillary hyperaemia, often in combination with the previous changes. Intermediate dose groups were also studied, and the changes were scored by "blind reading". Although difficult to determine by this way and based on data from only a limited number of animals, the lowest dose without effect could be estimated as 3 mg/kg for males and 10 mg/kg for females, or 3 mg/kg for sexes combined. The nodular hyperplasia, probably an indication of early neoplasia, was found in one animal of the 30 mg/kg group in either sex.
Morphometry was carried out for mitotic cells as a discrete parameter for the changes listed above. It can be concluded that a steep increase in mean number of mitotic cells was found with increasing dose, although the vatiation was considerable. ANOVA demonstrated a significant relation with dose, and Student-t revealed statistical significance (two-tailed) for groups treated with 3 and 10 mg/kg bw, respectively, when compared to controls.

THYMUS:
Thymus atrophy was diagnosed in relation to the structure of the thymic lobes, i.e. the relative quantity, shape and distribution of cortical and medullary areas. In animals displaying thymus atrophy, the proportion of medullary areas was increased with irregular boundaries. The incidence of this slight thymus atrophy in the 30 mg/kg groups appeared increased compared to the controls. For this reason also the intermediate groups were evaluated; this finding was recorded by blind reading. The incidence in these groups were nil or insignificantly low. Therefore the lowest dose without effect can be established at 10 mg/kg.

LIVER:
All animals contained a few to many small inflammatory cell foci consisting mainly of macrophages, occasionally surrounding a necrotic cell. When this lesion was quantified (categories 0-5; 5-10 and >10 foci per 2 liver sections), no difference between control and top dose group was apparent. In one male of both control and 30 mg/kg group a necrotic area was seen in the liver.
Slight fatty vacuolation was seen in a significant proportion of both males and females, with a slightly higher incidence in high dose males (when “vacuolation” and “fatty vacuolation” are combined). The distribution was more or less zonal, and occurred mainly in the periportal (zone I) areas. In a few females (1 in control, 2 in the top dose group) the vacuolation occurred more focally and was more of the glycogen type. This change was suggestive for early clear-cell foci.

LUNG:
Minimal inflammatory changes (inflammatory cell foci, histiocytosis, increased septal cellularity) occurred at a low incidence, although the number of animals affected was higher in the 30 mg/kg females (6/10 vs 2/10). This, as well as the arterial mineralisation observed is considered as a common background lesion.

OESOPHAGUS:
Changes in the oesophagus consisted of degeneration and regeneration of muscle fibers, and focal inflammation of the muscular wall. Although the covering mucosa appeared intact, these changes are considered as a result of blunt traumatic damage due to gavage application of the test material. These lesions occurred occasionally and were not related to test material exposure.

SPLEEN:
Occasional changes such as increased haemosiderin pigment, hyperaemia, lymphoid depletion or reticulum cell hyperplasia, were observed. Although these changes were found exclusively in treated animals, the incidence was too low to be considered significant.
No changes were observed in mesenteric lymph nodes and glandular stomach.
Remarks on result:
not measured/tested
Critical effects observed:
not specified
Conclusions:
This study identified the forestomach and thymus as target organs for toxicity with minimal effects at the higher dose levels. Consequently, there was no indication for a need to change the dose regimen for the subsequent carcinogenicity study.
Executive summary:

A sub-chronic repeated dose toxicity study was conducted to help inform on dose selection for a subsequent carcinogenicity study. This 90 -day study was conducted under GLP conditions.

During the study, groups of 10 animals per sex, per dose, received doses of test material, by oral gavage, for 5 days a week over a period of 90 days. The animals were observed daily and their behaviour and clinical signs were examined. Body weights, food and water consumption was measured at least weekly. Following the end of the exposure period the animals were sacrificed by exsanguination from the abdominal aorta. Blood, urine and tissues were taken for investigations on standard toxicological parameters. All animals were subjected to macroscopic examination, organ weights were recorded, and tissue samples were processed for further histopathological investigation. The purpose of this 90-day kill was to reassure the appropriateness of the dose levels chosen for the chronic carcinogenicity study.

In particular the forestomach was considered a target organ, in which ultimately the development of tumours was expected. A possible persistent irritation with frank hyperplasia induced by the compound would reduce the value of the tumour induction for extrapolation purposes, and possibly reduce the life span of the rats. Fortunately, the hyperplasia was only minimal and could be characterised as “basal cell disturbance”, and only in two out of 20 animals from the high dose group a small hyperplastic nodule was found, one of which showing dysplasia indicative of early neoplasia. However, due to the small size (no grossly observable changes) and the random sampling, this figure is probably underestimated. Further analysis of the basal cell hyperplasia was carried out by morphometric counting of BrdU incorporation. This confirmed the minimal changes observed by histopathology, and in addition provided data that could be analysed statistically by which a significant increase was found from 10 mg/kg onwards.

Another phenomenon was the atropy of the thymus, a confirmation of the reduction of thymus weight observed at necropsy, which occurred only at the top-dose.

Despite the clear increase in liver weight by the test material, examination of histopathology at the microscopic level, as well as of clinico-chemical parameters (ALAT, γGT, LDH) did not reveal any signs of hepatocellular toxicity.

In conclusion, this study identified the forestomach and thymus as target organs for toxicity with minimal effects at the higher dose levels. Consequently, there was no indication for a need to change the dose regimen.

Endpoint:
sub-chronic toxicity: oral
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Justification for type of information:
JUSTIFICATION FOR DATA WAIVING
The repeated dose toxicity data requirements of REACH are waivedon the basis of the provisions provided in Annex XI of the regulation, Section 1 (testing does not appear scientifically necessary) on the basis of section 1.1 “use of existing data” and section 1.2 “weight of evidence”. The following further outline the relevant justifications:

1. Consideration of the substance compositions and the hazard classifications of the constituent substances
The hazard classifications of the constituent substances of the shale oil fractions were assessed and none indicated the need to classify for specific target organ toxicity following repeated exposure (please refer to the document ‘Shale oils RAAF Report’, as included in section 13, for further information).

2. Consideration of the presence of PAH in each of the registered substances and the view that PAHs are the drivers for the long term toxicity hazards of the registered substances
Analytical information on the registered substances shows that PAHs are present at 1% in the registered substance. Benzo[a]pyrene was the only PAH identified (heavy fraction analysis). Bearing in mind the accepted opinion that PAHs can be grouped in view of the similarities in their toxicity profiles, and in view of the particularly stringent harmonised classification of benzo[a]pyrene, benzo[a]pyrene data are used as a surrogate for the PAH content of the registered substances. Available repeated dose toxicity data on benzo[a]pyrene, as generated by the Dutch authorities (RIVM, 2001), is therefore included in the dataset of the registered substance and is considered to provide conservative assessment of long term toxicological hazard of the registered substancs. The data are included to support the repeated dose toxicity waivers and have been added as part of the general weight of evidence approach. The available study data on benzo[a]pyrene are considered to be equivalent to data generated by corresponding test methods referred to in Article 13(3) since they were conducted on behalf of the Ducth authorities and are:
1) adequate for the purpose of classification and labelling and/or risk assessment;
2) adequate and reliable in terms of coverage of the key parameters foreseen to be investigated in the corresponding test methods referred to in Article 13(3);
3) the exposure durations were comparable to or longer than the corresponding test methods referred to in Article 13(3) if exposure duration is a relevant parameter; and
4) adequate and reliable documentation of the studies are provided.

3. Consideration of the OEL that has been utilised in generation of conservative DNELs
OEL (occupational exposure limit) values for benzo[a]pyrene have been established by the European Agency for Safety and Health. In this report the lowest OEL value for 8 hours exposure is 0.002 mg/m³. It is considered this value can be viewed as adequately protective to the registered substance since benzo[a]pyrene can be considereda conservative surrogate for the PAH component, which itself can be considered to be the most hazardous component.

Additional repeated dose toxicity data are therefore not considered to be required in order to generate a DNEL. Furthermore, this approach is more conservative than the Virtually Safe Dose (VSD) of 5 ng benzo[a]pyrene/kg bw/day based on the results from the RIVM 2-year rat study (and are comparable to VSD based on the 2-year mice study).
In summary, the risk assessment is performed based on the most toxicologically relevant component, benzo(a)pyrene. Classification and DNELs can be derived based on calculation from the OEL recognised at European level. Even at 1%, the presence of PAHs are still the only component that need to be taken into consideration in the derivation of DNELs, because comparison of existing reference doses and exposure limits show that for PAHs are 10000 lower than next lowest reference dose i.e. extrapolating the OEL for each oil fraction from the PAH content still results in the lowest OEL, than if were done based on the content of the remaining materials.

4. Consideration of CMR (category 1B) classification and the stringent operational controls and risk management measures implemented during use to mitigate risk
In view of the CMR category 1B classifications that have been assigned to the registered substance, together with conservative approach taken in classifying for other REACH endpoints, it is not considered to be an expedient use of animals to commission repeated dose toxicity studies on the registered substance. Comprehensive operational controls and risk management measures are implemented at sites using the substances, to ensure risk mitigation of the CMR (category 1B) hazard. The measures that are implemented to ensure safe use of a CMR category 1B substance are considered protective for any potential repeated dose toxicity hazard. Any information which might be generated from repeated dose toxicity studies would not provide data that would improve the safety of workers who might be exposed to the substances.
The exposure scenarios, as included in chapters 9 and 10 of the substance CSRs, cover all relevant exposures throughout the lifecycle of the substance and demonstrate that the substance is not incorporated into an article. In all cases, comprehensive OCs/RMMs are enforced to the extent that risk to humans and to the environment is mitigated.

In addition to the column 2 and Annex XI justifications outlined above. The waivers are further justified on exposure considerations and animal welfare grounds.

Exposure considerations
The level of potential exposure was modelled (using ART), taking account of the relevant operational controls (OCs) and risk management measures (RMMs) that are applied. In all cases, comprehensive OCs/RMMs are enforced to the extent that there is minimal opportunity for exposure during use of the substance to humans and to the environment.
All workers receive training in the handling of the substances and all workers are required to wear an appropriate level of personal protective equipment, as a secondary means of protection to the primary technical means of control. All personal protective equipment conforms to European Standards and are selected in consideration of the exposure period, the expected level of exposure and are made of the appropriate material/ have the appropriate Assigned Protection Factor (AF).
As part of a qualitative exposure assessment of human health, which was conducted as a means of providing an additional level of assurance on the lack of risk, the potential (worst case estimates for) exposures were compared to the relevant OELs. In all cases, the exposure values predicted for ‘shale oil’ are considerably less than the OELs that are used for monitoring purposes at the facilities using shale oil and, as such, the use of the substances are not considered to pose a risk to the health of workers.
There are no consumer uses of the substances.
Overall, no risk to human health is anticipated in view of the operational controls and risk management measures that are implemented. Furthermore, chemical monitoring programs at the use sites confirms that workers are not exposed to any of the substances at levels above the OELs.

Animal welfare considerations
The protection and welfare of animals is an area covered by a wide range of EU legislation. Replacement, Reduction and Refinement (the “Three Rs”) is the key strategy of a systematic framework aimed at achieving the goal of humane experimental techniques. In order to improve the welfare of animals still needed for scientific purposes, the aim is to firmly anchor the principle of the “Three Rs” in EU legislation. In this context, Regulation (EC) No 1907/2006, Article 25 (1), clearly states that animal testing is to be avoided and only allowed as a last resort. The use of existing data as part of a weight of evidence approach has indicated that the use of benzo[a]pyrene data can be considered appropriate and protective for the mitigation of risk following long-term and repeated exposure to the registered substances. Further testing with the registered substances is therefore not considered to be an expedient, or responsible, use of animals.

Please refer to "Justification for type of information" and also to the attached document '1904651.UK0 - 7899 Consideration of the long term toxicity of Distillates (shale oil)' for further information.
Limit test:
no

Additional information

The repeated dose toxicity data requirements of REACH are waived on the basis of the provisions provided in Annex XI of the regulation, Section 1 (testing does not appear scientifically necessary) on the basis of section 1.1 “use of existing data” and section 1.2 “weight of evidence”. The following further outline the relevant justifications:


 


1.       Consideration of the substance compositions and the hazard classifications of the constituent substances


The hazard classifications of the constituent substances of the shale oil fractions were assessed and none indicated the need to classify for specific target organ toxicity following repeated exposure (please refer to the document ‘Shale oils RAAF Report’, as included in section 13, for further information).


 


2.       Consideration of the presence of PAH in each of the registered substances and the view that PAHs are the drivers for the long term toxicity hazards of the registered substances


Analytical information on the registered substances shows that PAHs are present at 1% in the registered substance. Benzo[a]pyrene was the only PAH identified (heavy fraction analysis). Bearing in mind the accepted opinion that PAHs can be grouped in view of the similarities in their toxicity profiles, and in view of the particularly stringent harmonised classification of benzo[a]pyrene, benzo[a]pyrene data are used as a surrogate for the PAH content of the registered substances. Available repeated dose toxicity data on benzo[a]pyrene, as generated by the Dutch authorities, is therefore included in the dataset of the registered substance and is considered to provide conservative assessment of long term toxicological hazard of the registered substancs. The data are included to support the repeated dose toxicity waivers and have been added as part of the general weight of evidence approach. The available study data on benzo[a]pyrene are considered to be equivalent to data generated by corresponding test methods referred to in Article 13(3) since they were conducted on behalf of the Ducth authorities and are:


1) adequate for the purpose of classification and labelling and/or risk assessment;


2) adequate and reliable in terms of coverage of the key parameters foreseen to be investigated in the corresponding test methods referred to in Article 13(3);


3) the exposure durations were comparable to or longer than the corresponding test methods referred to in Article 13(3) if exposure duration is a relevant parameter; and


4) adequate and reliable documentation of the studies are provided.


 


3.       Consideration of the OEL that has been utilised in generation of conservative DNELs


OEL (occupational exposure limit) values for benzo[a]pyrene have been established by the European Agency for Safety and Health. In this report the lowest OEL value for 8 hours exposure is 0.002 mg/m³. It is considered this value can be viewed as adequately protective to the registered substance since benzo[a]pyrene can be considereda conservative surrogate for the PAH component, which itself can be considered to be the most hazardous component.


Additional repeated dose toxicity data are therefore not considered to be required in order to generate a DNEL. Furthermore, this approach is more conservative than the Virtually Safe Dose (VSD) of 5 ng benzo[a]pyrene/kg bw/day based on the results from the RIVM 2-year rat study (and are comparable to VSD based on the 2-year mice study).


In summary, the risk assessment is performed based on the most toxicologically relevant component, benzo(a)pyrene.  Classification  and DNELs can be derived based on calculation from the OEL recognised at European level. Even at 1%, the presence of PAHs are still the only component that need to be taken into consideration in the derivation of DNELs, because comparison of existing reference doses and exposure limits show that for PAHs are 10000 lower than next lowest reference dose i.e. extrapolating the OEL for each oil fraction from the PAH content still results in the lowest OEL, than if were done based on the content of the remaining materials.


 


4.       Consideration of CMR (category 1B) classification and the stringent operational controls and risk management measures implemented during use to mitigate risk


In view of the CMR category 1B classifications that have been assigned to the registered substance, together with conservative approach taken in classifying for other REACH endpoints, it is not considered to be an expedient use of animals to commission repeated dose toxicity studies on the registered substance. Comprehensive operational controls and risk management measures are implemented at sites using the substances, to ensure risk mitigation of the CMR (category 1B) hazard. The measures that are implemented to ensure safe use of a CMR category 1B substance are considered protective for any potential repeated dose toxicity hazard. Any information which might be generated from repeated dose toxicity studies would not provide data that would improve the safety of workers who might be exposed to the substances.


The exposure scenarios, as included in chapters 9 and 10 of the substance CSRs, cover all relevant exposures throughout the lifecycle of the substance and demonstrate that the substance is not incorporated into an article. In all cases, comprehensive OCs/RMMs are enforced to the extent that risk to humans and to the environment is mitigated.


 


In addition to the column 2 and Annex XI justifications outlined above. The waivers are further justified on exposure considerations and animal welfare grounds.


 


> Exposure considerations


The level of potential exposure was modelled (using ART), taking account of the relevant operational controls (OCs) and risk management measures (RMMs) that are applied. In all cases, comprehensive OCs/RMMs are enforced to the extent that there is minimal opportunity for exposure during use of the substance to humans and to the environment.


All workers receive training in the handling of the substances and all workers are required to wear an appropriate level of personal protective equipment, as a secondary means of protection to the primary technical means of control. All personal protective equipment conforms to European Standards and are selected in consideration of the exposure period, the expected level of exposure and are made of the appropriate material/ have the appropriate Assigned Protection Factor (AF).


As part of a qualitative exposure assessment of human health, which was conducted as a means of providing an additional level of assurance on the lack of risk, the potential (worst case estimates for) exposures were compared to the relevant OELs. In all cases, the exposure values predicted for ‘shale oil’ are considerably less than the OELs that are used for monitoring purposes at the facilities using shale oil and, as such, the use of the substances are not considered to pose a risk to the health of workers.


There are no consumer uses of the substances.


Overall, no risk to human health is anticipated in view of the operational controls and risk management measures that are implemented. Furthermore, chemical monitoring programs at the use sites confirms that workers are not exposed to any of the substances at levels above the OELs.


 


> Animal welfare considerations


The protection and welfare of animals is an area covered by a wide range of EU legislation.  Replacement, Reduction and Refinement (the “Three Rs”) is the key strategy of a systematic framework aimed at achieving the goal of humane experimental techniques.  In order to improve the welfare of animals still needed for scientific purposes, the aim is to firmly anchor the principle of the “Three Rs” in EU legislation.  In this context, Regulation (EC) No 1907/2006, Article 25 (1), clearly states that animal testing is to be avoided and only allowed as a last resort.  The use of existing data as part of a weight of evidence approach has indicated that the use of benzo[a]pyrene data can be considered appropriate and protective for the mitigation of risk following long-term and repeated exposure to the registered substances. Further testing with the registered substances is therefore not considered to be an expedient, or responsible, use of animals.


 


Please refer to "Justification for type of information" and also to the attached document '1904651.UK0 - 7899 Consideration of the long term toxicity of Distillates (shale oil)' for further information.


 


 


The supporting data on benzo[a]pyrene can be summarised, as follows. These data are included as part of a weight of evidence approach and are considered together with available data supporting the CMR (category 1B) classification of the registered substance, in view of its composition and its inclusion of PAHs at 1%.


 


Short-term repeated dose toxicity (oral): supporting data on benzo[a]pyrene


A short term repeated dose toxicity study was conducted to help inform on dose selection for a subsequent carcinogenicity study. This range-finding study was conducted under GLP conditions.


During the study, groups of 10 animals per sex, per dose, received doses of test material, by oral gavage, 5 days a week. The animals were observed daily and their behaviour and clinical signs were examined. Body weights, food and water consumption was measured weekly. During the sixth week of exposure the animals were killed by exsanguination under ether anesthesia and blood, urine and tissues were taken for investigations on standard toxicological parameters. All animals were subjected to macroscopical examination, organ weights were recorded, and tissue samples were processed for further histopathological investigations. In addition, liver enzyme induction was monitored by EROD (ethoxyresurfin-O-deethylase) activity in plasma.


On the basis of findings from this study, particularly in relation to effects noted on basal cell hyperplasia in the forestomach as well as the effects upon liver- and thymus-weight, doses of 3, 10 and 30 mg/kg bw/day were proprosed for the carcinogenicity study. In order to verify the observed forestomach proliferation, morphometrical analysis of the forestomach was incorporated into a 90-day repeated dose toxicity stduy in rats using the BrdU-incorporation technique (see a robust study summary under section 7.5.1). This study essentially showed similar results.


 


Sub-chronic repeated dose toxicity (oral): supporting data on benzo[a]pyrene


A sub-chronic repeated dose toxicity study was conducted to help inform on dose selection for a subsequent carcinogenicity study. This 90 -day study was conducted under GLP conditions.


During the study, groups of 10 animals per sex, per dose, received doses of test material, by oral gavage, for 5 days a week over a period of 90 days. The animals were observed daily and their behaviour and clinical signs were examined. Body weights, food and water consumption was measured at least weekly. Following the end of the exposure period the animals were sacrificed by exsanguination from the abdominal aorta. Blood, urine and tissues were taken for investigations on standard toxicological parameters. All animals were subjected to macroscopic examination, organ weights were recorded, and tissue samples were processed for further histopathological investigation. The purpose of this 90-day kill was to reassure the appropriateness of the dose levels chosen for the chronic carcinogenicity study.


In particular the forestomach was considered a target organ, in which ultimately the development of tumours was expected. A possible persistent irritation with frank hyperplasia induced by the compound would reduce the value of the tumour induction for extrapolation purposes, and possibly reduce the life span of the rats. Fortunately, the hyperplasia was only minimal and could be characterised as “basal cell disturbance”, and only in two out of 20 animals from the high dose group a small hyperplastic nodule was found, one of which showing dysplasia indicative of early neoplasia. However, due to the small size (no grossly observable changes) and the random sampling, this figure is probably underestimated. Further analysis of the basal cell hyperplasia was carried out by morphometric counting of BrdU incorporation. This confirmed the minimal changes observed by histopathology, and in addition provided data that could be analysed statistically by which a significant increase was found from 10 mg/kg onwards.


Another phenomenon was the atropy of the thymus, a confirmation of the reduction of thymus weight observed at necropsy, which occurred only at the top-dose.


Despite the clear increase in liver weight by the test material, examination of histopathology at the microscopic level, as well as of clinico-chemical parameters (ALAT, γGT, LDH) did not reveal any signs of hepatocellular toxicity.


In conclusion, this study identified the forestomach and thymus as target organs for toxicity with minimal effects at the higher dose levels. Consequently, there was no indication for a need to change the dose regimen.

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 repeated dose toxicty.