Registration Dossier

Diss Factsheets

Administrative data

Description of key information

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:
From 2008-08-25 to 2008-12-10
Reliability:
1 (reliable without restriction)
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
other: OFA
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles Rivers Laboratories (Wilmington, MA)
- Age at study initiation: 8-10 weeks
- Weight at study initiation: 180-250g
- Fasting period before study: Animals were fasted overnight prior to the test item administration and food returned at 4h post dose. Water was available ad libitum with no restriction.
- Housing: The rats were housed multiply in cages (group-caged per dose) in the animal facilities of CER-DHA.
- Diet : ad libitum
- Water: ad libitum
- Acclimation period: At least 5 days in the experimental unit before use in the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.5 ± 5.5
- Humidity (%): 45-90
- Air changes (per hr): 10-30
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- For steps 1 and 2 of the procedure (administration of the test item at a final dose of about 300 mg/kg bw), the administrated solution was prepared by dissolving 10x the test item in corn oil in order to obtain an adequate administration volume. The administrated solution was prepared in the hour preceding administration to animals and stored at room temperature.
- For steps 3 and 4 (administration of the test item at a final dose of about 2000 mg/kg bw), the test item was directly administrated to animals without dissolution in any vehicle.
- Lot/batch no. (if required): 117K0127


MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: since no information about toxicity of the test item was available, the chosen starting dose level was 300 mg/kg bw as recommended by OECD guideline for animal welfare.
Doses:
Steps 1 and 2 of the procedure: administration of the test item at a final dose of about 300 mg/kg bw
Steps 3 and 4: administration of the test item at a final dose of about 2000 mg/kg bw
No. of animals per sex per dose:
2 groups of 3 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: At least once during the first 30 minutes, periodically during the first 24h, with special attention given during the first 4 hours, and daily thereafter, for a total of 14 days.
- Frequency of weigthings: at the beginning of the test
- Necropsy of survivors performed: yes
- Other examinations performed: For animals surviving 24 hours or more, a microscopic examination of organs showing evidence of gross pathology was also considered.
Statistics:
None
Preliminary study:
Not relevant
Sex:
female
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: According to the experimental scheme recommended by the OECD guideline 423, the test item can be classified in the category 5 of the GHS acute toxicity scheme which corresponds to a limit LD50 higher than 5000 mg/kg bw.
Mortality:
Step 1: 0/3
Step 2: 0/3
Step 3: 0/3
Step 4: 0/3
Clinical signs:
None
Body weight:
Not measured (except at the beginning of the experiment)
Gross pathology:
None
Other findings:
None
Interpretation of results:
Category 5 based on GHS criteria
Remarks:
Migrated information
Conclusions:
From these experiments and on the basis of the followed experimental scheme recommended by OECD TG 423, it can be concluded that TOU can be classified in the category 5 of the GHS acute toxicity scheme which corresponds to a limit LD50 higher to 5000 mg/kg bw for this test item.
Executive summary:

The objective of this study was to investigate the acute toxicity of the chemical compound 2,5,7,10 -tetraoxaudecane (TOU). In order to assess the toxicity of TOU, the method of predefined doses (acute toxic class method) was chosen. Ultimately, the method allows the tested substance to be ranked and classified according to the Globally Harmonised System (GHS) for the classification of chemicals which cause acute toxicity.

The acute toxic class method is a stepwise procedure based on biometric evaluations with fixed doses, adequately separated to enable a substance to be ranked for classification purposes and hazard assessment.

As recommended by the OECD TG 423, in a first step, TOU (mixed with corn oil) was administrated at a final dose level of about 300 mg/kg bw. Following administration, animals were regularly inspected for tolerance to the tested chemical compound administration. From the TOU administration time (T) to the exposure time T+4h, 9 to 10 observations were performed and recorded for each animal. The next observations, from D+2 (day ensuing the administration day D+1) to D+14 were performed daily with a frequency of 1 inspection point per day. After an observation period of 14 days, all 3 animals were alive without evident toxicological effect apparitions resulting from administration of TOU. Finally, animals were humanely killed by 1ml IP injection and macroscopically autopsied. All tissues were examined with a specific attention to hearth, liver, kidney, lungs, spleen, brain and stomach. As a result, no obvious pathological tissue alterations resulting from TOU oral administration were observed. Nevertheless, some lesions (especially pulmonary and hepathic congestion) probably caused by Nembutal anaesthesia. Indeed it is known that barbiturate induce in particular respiratory depression and are metabolised by the liver. Finally, all take in organs were individually identified and stored in formaldehyde solutions for further histological analyses if necessary.

In a second step, the initial administrated dose (about 300 mg/kg) was once again orally injected to 3 other female rats in order to confirm the results obtained in the first step. From the TOU administration time (T) to the exposure time T+4h, 10 observations were performed and recorded for each animal. The next observations, from D+2 (day ensuing the administration day D+1) to D+14 were performed daily with a frequency of one inspection point per day. From this second injection, after an observation period of 14 days, the same observations on those highlighted for the step 1, namely, no mortality in the test animal group and no evident external and internal pathological alterations resulting from administration of a TOU solution were highlighted. After anaesthesia (1ml Nembutal), rats were autopsied and sampled tissues (hearth, liver, kidneys, lungs, brain and stomach) were examined.

Results of the second step led to the following conclusion: administration of a TOU solution at a final dose volume of 300 mg/kg bw induces no acute toxicity in female rats.

Since no death and no toxicological effects were observed after a 300 mg/kg administration, TOU was administrated at a higher dose to 3 other female rats. TOU was administrated at a final oral dose of 2000 mg/kg bw as recommended by OECD TG 423. As for the steps 1 and 2, following oral administration, animals were carefully observed (6 observations for day D+1 and daily thereafter) for any toxicological adverse effects as described previously. Like for administration of TOU at 200 mg/kg bw, no death or toxicological adverse effect apparitions which could result from the TOU exposure were highlighted for a 14 days period following a first administration of TOU at 2000 mg/kg bw. Autopsies revealed no obvious pathological tissue alterations,except those probably caused by Nembutal, were highlighted for all tested animals. Sampled tissues (hearth, liver, kidneys, lungs, spleen, brain and stomach) were also conserved in appropriate media.

Repetition of the same exposure conditions (about 2000 mg/kg bw) to 3 other female rats and same observation period led to the same final observations and conclusions to that described for the first administration of a dose level at about 2000 mg/kg bw.

From these experiments and on the basis of the followed experimental scheme recommended by OECD TG 423, it can be concluded that TOU can be classified in the category 5 of the GHS acute toxicity scheme which corresponds to a limit LD50 higher to 5000 mg/kg bw for this test item.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
5 000 mg/kg bw
Quality of whole database:
The test was performed according to OECD TG 423 and GLP (Klimisch code 1).

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
04. 06. – 21. 06. 2012
Reliability:
1 (reliable without restriction)
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: Wistar Han
Sex:
male/female
Details on test animals or test system and environmental conditions:
Species: Laboratory rat
Strain: Wistar Han, monitored quality (quality status guaranteed by supplier)
Source: Breeding farm VELAZ s.r.o., Koleč u Kladna, Czech Republic, RČH CZ 21760152
Sex: Males and females
Acclimatisation: 5 days
Total number: 5 males + 5 females
Housing: animal room with monitored conditions – one animal in one plastic cage
Diet: ST 1 BERGMAN – standard pelleted diet ad libitum (producer: Ing.Mrkvička Miroslav - Výroba krmných směsí, Mlýn Kocanda, 252 42 Jesenice u Prahy).
Water: drinking tap water ad libitum (quality corresponding to the Regulation No. 252/2004 of Czech Coll. of Law)
Microclimatic conditions: room temperature 22 + 3 °C, permanently monitored relative humidity 30 – 70 %, permanently monitored light: 12 hour light/12 hour dark
Bedding: sterilized shavings of soft wood
Identification of animals: colour marks on tail, each cage was marked with the number of study, number of animal, sex, name and dose of the test
substance.
Randomisation: according to the internal rule, at the start of the study the weight variation of animals was minimal and did not exceed + 20 % of the mean weight for each sex.
Health condition: certificate of good health condition – from breeding farm; no signs of diseases were observed at clinical check-in, during
the acclimatisation period and before the start of study.
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
Preparation of experimental animals
Approximately 24 hours before application, a skin area of about 6 x 6 cm on the back of animals was shaved (aprox. 10% of the body surface). The animals were weighed immediately before application.

Preparation and application of the test substance
The amount of test substance for each animal was weighed out (according to its body weight and the dose) immediately before application.
The test substance in delivered form was applied on the depilated area of skin. The application site was covered by mull and held in contact by plaster (strapping). After 24 hours the semiocclusive dressing was removed, and remains of the test substance were wiped off with water.
Duration of exposure:
14 days
Doses:
The study was performed as limit test: two groups of animals – 5 males and 5 females and the dose of 2000 mg/kg. The pre-test was performed with 1 male and 1 female of the groups of the five animals. The pre-test was started one day before the start of limit test.
No. of animals per sex per dose:
5
Control animals:
not required
Details on study design:
Time schedule of observations
Body weight: before application, 8th and 15th day of study
Mortality: daily
Clinical signs: daily
Pathological examination: 15th day of study

Body weight recording
The animals were weighed at the start of the study (before application), at 8th day and at the end of experiment (15th day). The average body weight of the groups was calculated from individual body weights.
Body weight increments were calculated from the body weight at the start of the study, during the first week and at the end of the study (see table No. 1 and No. 2).

Clinical observation
After application the animals were observed individually – at the first day: twice (30 minutes and 3 hours after application), at the second day: twice (in the morning and in the afternoon) and daily thereafter for 14 days.
Observations included changes in skin and fur, eyes, visible mucous membranes, nutritive condition, autonomic and central nervous systems, psychic activity, somatomotor activity, reactions to stimuli, function of respiratory, circulatory, digestive and urogenital system.
Also presence of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma was carefully observed.

Pathological examination
All test animals survived to the end of study were sacrificed on the 15th day by injection of veterinary preparation T61 (1 ml iv.) and gross necropsy was carried out. Nutritious status, body surface, body foramina, thoracic, abdominal and cranial cavity were evaluated. All gross macroscopic changes of organs and tissues were recorded on special data sheets.
Preliminary study:
not relevant
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No death was detected after application in all animals.
Clinical signs:
No clinical signs of intoxication were detected after application in all animals.
Body weight:
In males and females, the weight increments were adequate to species and age of animals in the experiment with average weights of:
- 260.67 g before application, 286.97 g at day 8 and 327.19 g at day 15 for males
- 222.52 g before application, 229.53 g at day 8 and 242.95 g at day 15 for females
Gross pathology:
No macroscopic changes were diagnosed during pathological examination in all animals
Other findings:
None
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Evaluation of results
The test substance toxicity was evaluated on the basis of mortality, body weight changes, and clinical signs of toxicity during the observation period and necropsy findings at the end of study.
The test substance applied on skin at a dose of 2000 mg/kg of animal weight did not cause death of animals.
No clinical signs of intoxication were observed. No macroscopic changes were diagnosed during pathological examination of animals.

Test substance toxicity characterization
According to the results of study, the value of LD50 (dermal) of the test substance, 2,5,7,10-TETRAOXAUNDECANE (TOU), is higher than 2000 mg/kg of body weight for rats of both sexes.
Executive summary:

The test substance, 2,5,7,10-TETRAOXAUNDECANE (TOU), was tested for acute dermal toxicity using Wistar rats. Testing was performed according to the Method B.3 Acute Toxicity (Dermal), Council Regulation (EC) No.440/2008, published in O.J. L 142, 2008.

The study was performed as limit test: two groups of animals – 5 males and 5 females at the dose of 2000 mg/kg of body weight. The pre-test was performed with 1 male and 1 female from each group. After end exposure test substance of these pilot animals, the other animals of the group were dosed. The test substance was applied on the shaved skin of the test animals in delivered form for 24 hours. The test animals were observed 14 days after exposure test substance, afterwards they were sacrificed, and the necropsy for macroscopic examination of the organs was performed.

The test substance applied at the dose of 2000 mg/kg of body weight did not cause death of animals. No clinical signs of intoxication were observed. No macroscopic change was diagnosed during pathological examination of animals.

According to the results of study, the value of LD50 (dermal) of the test substance, 2,5,7,10-TETRAOXAUNDECANE (TOU), is higher than 2000 mg/kg of body weight for rats of both sexes.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
The test was performed according to OECD TG 402 and GLP (Klimisch code 1).

Additional information

Justification for selection of acute toxicity – oral endpoint

The test was selected as the only one available.

Justification for selection of acute toxicity – inhalation endpoint

In accordance with column 2 of REACH Annex VIII relative to Acute toxicity (Point 8.5), in addition to the oral route (8.5.1), for substances other than gases, the information mentioned under 8.5.2 (acute toxicity by inhalation) to 8.5.3 (acute toxicity by derma route) shall be provided for at least one other route. The choice for the second route will depend on the nature of the substance and the likely route of human exposure. If there is only one route of exposure, information for only that route need be provided.

Testing by dermal route is appropriate regarding to the test substance exposure and physicochemical and toxicological properties.

Testing by inhalation is not appropriate.

Justification for selection of acute toxicity – dermal endpoint

The test was selected as the only one available.

Justification for classification or non-classification

The test substance does not meet the criteria for being classified as dangerous as no mortality/effect has been identified up to the limit test in oral and inhalation acute toxicity tests.

Categories Display