Registration Dossier

Administrative data

Description of key information

Acute oral toxicity: The LD50 value for renewable hydrocarbons of wood origin is 2000 mg/kg bw .
Acute dermal toxicity: The LD50 for read-across substance is 40000 mg/kg bw.
Acute inhalation toxicity: The LC50 value for read-across substance is 23400 mg/m3

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:
26 April 2012 - 17 may 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to the OECD 420 guideline and in compliance with GLP.
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Deviations:
no
GLP compliance:
yes
Test type:
fixed dose procedure
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: harlan Laboratories UK Ltd., Oxon, UK
- Age at study initiation: eight to twelve weeks
- Weight at study initiation: 155-167 g
- Fasting period before study: overnight fast immediately before dosing and for approximately three to four hours after dosing
- Housing: in groups of up to four in suspended solid-floor polypropylene cages furnished with woodflakes
- Diet (e.g. ad libitum): 2014C Teklad Global Rodent diet ad libitum
- Water (e.g. ad libitum): drinking water ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): at least 15 per hour
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
Using available information on the toxicity of the test item, one animal was gavaged at a dose level of 2000 mg/kg. In the absence of toxicity, an additional group of animals (four) were gavaged at 2000 mg/kg bw. All animals were dosed once using metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to its fasted bodyweight at the time of dosing.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were made 0.5, 1, 2 and 4 hours after dosing and subsequently once daily for fourteen days. Morbidity and mortality checks were made twice daily. Individual bodyweights were recorded on Day 0 (the day of dosing) and on Days 7 and 14.
- Necropsy of survivors performed: yes/no
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:
Preliminary study:
2000 mg/kg bw was chosen as the starting dose, no mortality was observed.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
There were no deaths (See table 1)
Clinical signs:
No signs of systemic toxicity were noted (See table 1)
Body weight:
All animals showed expected gains in bodyweight over the observation period (See table 2)
Gross pathology:
No abnormalities were noted at necropsy (See table 3)
Other findings:
- Organ weights: not measured
- Histopathology: not examined
- Potential target organs: not observed
- Other observations: none
Interpretation of results:
study cannot be used for classification
Remarks:
Migrated information
Conclusions:
The acute oral medial lethal dose (LD50) of the test item in the female rat was estimated to be greater than 2000 mg/kg bodyweight.
Executive summary:

The objective of this study was to assess the toxicity of the test item when administered in a single oral dose (2000 mg/kg bw) to female rats. No mortalities were observed at the tested dose level.

The study is considered reliable without restrictions since the study is carried out based on the OECD No. 420 guideline and in compliance with principles of Good Laboratory Paractice (GLP).

Based on these results, the test item does not have to be classified and has no obligatory labeling requirement for acute oral toxicity according to CLP Regulation 1272/2008 and Directive 67/548/EEC.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
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:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The study does not comply fully with the specific testing guideline. However, the study is well documented and the method used is scientically acceptable. The information in the publication is used to support the waiver. Read-across justification: Based on the chemical composition, the renewable hydrocarbons produced from raw materials such as fatty acid rich oil like Crude Tall Oil (CTO) or triglyserides, using a hydrotreatment process have similar hydrocarbon fractions and they contain the same critical constituents than fossil diesel fuels. According to the identified hydrocarbon blocks, the typical carbon number ranges and the physicochemical properties, the renewable hydrocarbons with diesel type fractions can be considered as having structural similarities and similar behaviour in contact with water and in the physiological processes than the analogue source substances (fossil diesel fuels). Their irritation properties, skin sensitisation property as well as acute and long-term adverse effects to human health is similar. Therefore, and in order to avoid the unnecessary animal testing, the read-across data from the analogue fossil diesel fuels is used to evaluate skin and eye irritation, the genetic toxicity, carcinogenicity, developmental toxicity and short term and/or long-term toxicological effects of the target substance.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Principles of method if other than guideline:
Male Sprague Dawley rats were exposed to inhaltion of n-C9 to n-C13 alkanes close to air saturation at 20 deg. C (4438, 1369, 442, 142 and 41 ppm, respectively) for 8 hours and observed for the following 14 days. In addition, exposure to higher and lower concentrations of n-C9 alkane was performed.
GLP compliance:
not specified
Test type:
other: Modification of the standard method
Species:
rat
Strain:
Sprague-Dawley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Mollegaard A/S, Ll. Skensved, Denmark
- Weight at study initiation: 180-220 g
- Acclimation period: 4-6 d
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-23
- Humidity (%): 40-70
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:
inhalation: vapour
Type of inhalation exposure:
whole body
Vehicle:
clean air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: conical 0.7 m3 steel chambers with glass front door and walls; flow rate 30-40 l/min.
- Vapour generation: test substance warmed (approx. 22 degrees C) under negative pressure and mixed with clean air to achieve required concentration
- Method of particle size determination:The presence of alkane aerosol in the inhalation chamber during exposure was investigated by Royco mod. 255 particlem onitorwith an aerosol particle counter sensor mod 241
- Temperature, humidity, pressure in air chamber: slightly higher than in the vapour generation, negative pressure 2-5 mm H20.

TEST ATMOSPHERE
- Brief description of analytical method used:Concentration of alkanes in the chambers was monitored automatically every 15 min by on-line gas chromatography (Shimadzu GC-9A).Chromatography was performed at 200 deg C on a 2 m x 1/8" stainless steel column packed with GP 10% SP-2100 on Supelcoport 100/120 mesh with helium as the carrier gas. The alkanes were detected by a FID detector. Injector and detector temperatures were 250 deg C.

TEST ATMOSPHERE (if not tabulated)
- Particle size distribution:
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.):
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
8 h
Concentrations:
n-C9: 5280*, 4438, 3560 and 2414 ppm
n-C10: 1369* ppm
n-C11: 442* ppm
n-C12: 142* ppm
n-C13: 41* ppm
Values marked * were the maximum achievable vapour concentration in air
No. of animals per sex per dose:
10 males
Control animals:
no
Details on study design:
- Duration of observation period following administration: Animals were exposed to at least the saturated vapour concentration (20 degrees C) for each test substance for 8 hr.
- Frequency of observations and weighing: During inhalation all animals were observd at 15 and 30 min intervals. For the first 8 hours after exposure all animals were observed hourly, then at 2 hour intervals during daytime for 14 days.
- Necropsy of survivors performed: yes, full gross necropsy on day 14
- Other examinations performed: clinical signs, histopathology, body weight, tissue weights of brain, lungs, heart, liver and kidney, the LC50 for n-nonane was calculated by probit analysis.
Sex:
male
Dose descriptor:
LC50
Effect level:
4 467 ppm
Based on:
test mat.
Remarks:
n-nonane
Exp. duration:
8 h
Remarks on result:
other: 23.4 mg/l
Sex:
male
Dose descriptor:
LC0
Effect level:
1 390 ppm
Based on:
test mat.
Remarks:
n-decane
Exp. duration:
8 h
Remarks on result:
other: 8.7 mg/l
Sex:
male
Dose descriptor:
LC0
Effect level:
442 ppm
Based on:
test mat.
Remarks:
n-undecane
Exp. duration:
8 h
Remarks on result:
other: 2.7 mg/l
Sex:
male
Dose descriptor:
LC0
Effect level:
142 ppm
Based on:
test mat.
Remarks:
n-dodecane
Exp. duration:
8 h
Remarks on result:
other: 0.99 mg/l
Sex:
male
Dose descriptor:
LC0
Effect level:
41 ppm
Based on:
test mat.
Remarks:
n-tridecane
Exp. duration:
8 h
Remarks on result:
other: 0.31 mg/l
Mortality:
n-nonane: 0/10, 1/10, 4/10, 9/10 at 2414, 3560, 4438 and 5280 ppm, respectively
n-decane, n-undecane, n-dodecane, n-tridecane: no mortality at maximum saturated vapour concentration
Clinical signs:
other: Tremor, spasms and limb paralysis were observed in rats treated with n-nonane. Most severe symptoms were observed in a high dose group. These symptoms were observed at 2 hours after the start of the exposure in the highest dose group. The length of time b
Body weight:
Not reported in the publication
Gross pathology:
No gross pathological changes were observed in the brain, lungs, heart, liver and kidneys following exposure to the n-C10 to n-C13 alkanes. In animals exposed to 4438 ppm of n-nonane the autopsy findings suggest that death was due to cardiopulmonary insufficiency. This may have been induced by the direct toxic effect of heart or lungs or by indirect effects by the central nervous system. In cerabellar cortex severe damage and extensive loss of purkinje neurons were demontrated.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute inhalation LC50(rat, male) for n-nonane was 4467 ppm (23.4 mg/l) following 8-hr exposure.
Executive summary:

The acute inhalation toxicity of a series on C9-C13 n-alkanes were investigated in male SD rats following a single 8 -hr exposure to vapour. A calculated LC50 of 4467 ppm (23.4 mg/l) was determined for n-nonane; the 4 -hr LC50 value is likely to exceed this value. No mortality was recorded for the other test substances after exposure to the maximum achievable vapour concentration in air. Maximum saturated vapour concentration decreased in a predictable manner with increasing C-number.

The study was conducted for the read-across substance. This study is considered reliable since the publication contains sufficiently data for assessment.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
Value:
23 400 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:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
This study is considered reliable with restrictions since the publication is well reported and contains sufficiently data for assessment, but is conducted for read-across data. Read-across justification: Based on the chemical composition, the renewable hydrocarbons produced from raw materials such as fatty acid rich oil like Crude Tall Oil (CTO) or triglyserides, using a hydrotreatment process have similar hydrocarbon fractions and they contain the same critical constituents than fossil diesel fuels. According to the identified hydrocarbon blocks, the typical carbon number ranges and the physicochemical properties, the renewable hydrocarbons with diesel type fractions can be considered as having structural similarities and similar behaviour in contact with water and in the physiological processes than the analogue source substances (fossil diesel fuels). Their irritation properties, skin sensitisation property as well as acute and long-term adverse effects to human health is similar. Therefore, and in order to avoid the unnecessary animal testing, the read-across data from the analogue fossil diesel fuels is used to evaluate skin and eye irritation, the genetic toxicity, carcinogenicity, developmental toxicity and short term and/or long-term toxicological effects of the target substance.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Principles of method if other than guideline:
Groups of five male and five female mice were administered a single 0.5-ml (dermal application of 5,000, 10,000,20,000, 30,000, or 40,000 mg/kg marine diesel fuel in 96% ethanol to the clipped dorsal interscapular region.
Test type:
other: modification of the standard method
Species:
mouse
Strain:
B6C3F1
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding: Laboratories
- Age at study initiation: 9 weeks
- Housing: Mice were housed five per cage
- Diet (e.g. ad libitum): feed ad libitum
- Water (e.g. ad libitum): water (acidified to pH 2.5) ad libitum
- Acclimation period: for 4 weeks before the studies began

ENVIRONMENTAL CONDITIONS
- Temperature (°F): 74" +/- 2
- Humidity (%): 30-70 %
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:
not specified
Vehicle:
ethanol
Remarks:
40 000 mg/kg dose applied as the neat chemical
Details on dermal exposure:
TEST SITE
clipped dorsal interscapular region.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): not done

TEST MATERIAL
- Amount(s) applied (volume): 0.5 ml
- Constant volume: yes

VEHICLE
- ethanol purity 95 %
Duration of exposure:
Single dose, no washing of the substance performed.
Doses:
5,000, 10,000,20,000, 30,000, or 40,000 mg/kg marine diesel fuel in 96% ethanol to the clipped dorsal interscapular region.
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: daily, weighed on day of dosing
- Necropsy of survivors performed: no
- Other examinations performed: clinical signs
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 40 000 mg/kg bw
Based on:
test mat.
Mortality:
not observed
Clinical signs:
no compound related signs of toxicity
Body weight:
not measured during the study
Gross pathology:
not performed
Other findings:
not reported
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute dermal LD50(mouse, male/female) for marine diesel fuel is 40 000 mg/kg/day after single exposure.
Executive summary:

The acute dermal toxicity of marine diesel fuel was investigated in male and female mice following a single dermal application of the test substance. A LD50 of 40 000 mg/kg/day was determined for the diesel marine oil; the highest concentration tested. No mortality was recorded for the test substances at any dose studied.

The study was conducted for the read-across substance. This study is considered reliable with restrictions since the publication is well reported and contains sufficiently data for assessment.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
40 000 mg/kg bw

Additional information

Oral exposure

In a key study of Sanders, A. (OECD 420 - Fixed dose method) conducted in accordance with GLP, one female rat was gavaged with undiluted test substance at a dose level of 2000 mg/kg bw. Following a sighting study test at dose level of 2000 mg/kg, an additional four fasted female animals were given a single oral dose level of 2000 mg/kg bodyweight. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy. The post exposure period was 14 days. There were no deaths observed during the study. Hunched posture and pilo-erection were noted in one animal during the day of dosing. All animals showed expected gains in bodyweight. No abnormalities were noted at necropsy. The acute median lethal dose (LD50) of the test item in female Wistar rat was estimated to be greater than 2000 mg/kg bodyweight.

As a conclusion, the results of the key study did not indicate this substance to be classified for acute toxicity via oral route.

Inhalation exposure

No valid data from renewable hydrocarbons of wood origin (diesel type fraction) are available on acute inhalation toxicity. The UVCB substance is currently on pilot production phase and no representative sample is currently available.

The vapour pressure of the substance is measured to be <0.1 kPa at 40 deg. C (method DIN EN 13016-1) indicating that the substance is not very volatile at ambient T. Thus, exposure via inhalation route is not significant. Based on the exposure assessment no high peak exposures are expected and the risks for acute effects are adequately controlled with the risk management measures presented for long-term effects (see CSR sections 9&10).

Therefore, unnecessary animal testing was considered unjustified and the hazard evaluation via inhalation route is based on the main constituents of the substance, alkanes C9 -C13 (Nilson et. al. 1988). Male rats were exposed to inhalation of each alkane close to air saturation at 20 deg. C for 8 hours and observed for 14 days. In addition, exposure to several concentration of C9 alkane (nonane) was performed. A calculated LC50 of 4467 ppm (23.4 mg/l) was determined for n-nonane; the 4 -hr LC50 value is likely to exceed this value. No mortality was recorded for the other test substances after exposure to the maximum achievable vapour concentration in air.

The aspiration property relates to the physico-chemical properties of the substance. Kinematic viscosity is a key parameter in determining aspiration hazards. Low viscosity substances have high potential to enter directly into trachea and lower air ways. The classification criteria for aspiration hazard of hydrocarbons refer to kinematic viscosity at 40 °C. The kinematic viscosities of the substance varied between 2.58 mm2/s to 2.77 mm2/s measured in accordance with DIN EN ISO 3104. Hydrocarbons with kinematic viscosity lower than 20.55 mm2/s should be classified for aspiration hazard.

Based on the above information there is no need to classify this substance as having acute toxicity via inhalation route. However, this substance is causing aspiration toxicity based on the kinematic viscosity.

Dermal exposure

The substance is a UVCB with varying composition. No representative sample is currently available from the substance and the animal testing is concluded to be not warranted by the rules laid down in REACH Annex XI.

Based on the exposure assessment no high peak exposures are expected and the risks for acute effects are adequately controlled with the risk management measures presented for long-term effects. When there is no data on the substance itself the read-across data was also used to avoid unnecessary animal testing. The read-across from fossil diesel fuels is justified because the toxicokinetics and toxicological properties of renewable diesel are considered similar to fossil diesel fuels based on the similar composition and physical-chemical properties. The read-across justification and the data matrices are presented in annex 1 of the CSR.

In a study of NTP, 1986 (equivalent or similar to OECD 402) with acceptable restrictions (no body weights measured during the study and no gross pathology performed at the end of the study) 5 male and 5 female mice were treated with the read-across substance (marine diesel oil) at concentrations 5 000, 10 000, 20 000, 30 000 and 40 000 mg/kg bw. The test substance was applied on unharmed skin of the dorsal interscapular region. Clinical signs were monitored during the study. The post exposure period was 14 days. There were no deaths observed during the study. The acute dermal LD50 value of read-across substance in male and female mice was greater than 40 000 mg/kg bw.

Based on the above information there is no need to classify this substance as having acute toxicity via dermal route.

 


Justification for selection of acute toxicity – oral endpoint
The study was conducted according to the OECD 420 guideline and in accordance with GLP.

Justification for selection of acute toxicity – inhalation endpoint
No study available for the target substance. This reliable study was conducted for read-across substance.

Justification for selection of acute toxicity – dermal endpoint
No study available for the target substance. This reliable study was conducted for read-across substance.

Justification for classification or non-classification

The available data for the substance and from the read-across substance, marine diesel oil, indicate relatively low potential for acute toxicity. Based on the available acute toxicity data, the substance has not to be classified according to CLP Regulation 1272/2008 and Directive 67/548/EEC.

Classification and labelling for aspiration toxicity is based on the measured kinematic viscosity of a substance at 40 °C. The measured kinematic viscosity of the test substance varies between 2.58 mm2/s to 2.77 mm2/s. Based on these values the substance is classified for Asp. Tox 1 H304 according to CLP Regulation 1272/2008 and for Xn; R65 according to Directive 67/548/EEC.