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EC number: 208-291-8 | CAS number: 520-34-3
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
![](https://chesar.echa.europa.eu/o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_toxicological-information.png)
Endpoint summary
Administrative data
Description of key information
Skin sensitisation: Key study. Method according to OECD 422B, GLP study. The Stimulation Indexes were 1.04, 1.01, and 0.84 at concentrations of 10, 25 and 50% test item, respectively. All values were below 1.6. Therefore, the test item has no sensitisation potential.
Key value for chemical safety assessment
Skin sensitisation
Link to relevant study records
- Endpoint:
- skin sensitisation: in vivo (LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- From November 16th to November 29th, 2016.
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 442B (Skin Sensitization: Local Lymph Node Assay: BrdU-ELISA)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of study:
- mouse local lymph node assay (LLNA): BrdU-ELISA
- Species:
- mouse
- Strain:
- CBA:J
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Elevage Janvier Labs (F-53941 Le Genest Saint Isle).
- Females nulliparous and non-pregnant: yes.
- Age at study initiation: 8 or 9 weeks old.
- Weight at study initiation: average weight 22.1 g (SD: 1.8)
- Housing: The animals were individually housed in suspended solid-floor polypropylene cages furnished with softwood woodflakes.
- Diet (e.g. ad libitum): tap water from public distribution system, ad libitum. Microbiological and chemical analyses of the water were carried out once every six months by Bureau Veritas – Eurofins (FRANCE).
- Water (e.g. ad libitum): Tkelad Global 18% Protein Rodent Diet (ENVIGO 2016) ad libitum.
- Acclimation period: at least 5 days.
- Indication of any skin lesions: no.
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19°C to 25°C
- Humidity (%): 30% to 70%
- Air changes (per hr): at least 10.
- Photoperiod (hrs dark / hrs light): 12 hrs dark / 12 hrs light. - Vehicle:
- dimethylformamide
- Concentration:
- 50%, 25% and 10%.
- No. of animals per dose:
- 4
- Details on study design:
- PRE-SCREEN TESTS: a preliminary screening test was performed using one mouse. The mouse was treated by daily application of 25 µL of the test item diluted at 50% in dimethylformamide (DMF) to the dorsal surface of each ear for three consecutive days (Days 1, 2, 3). The mouse was observed daily from day 1. Any signs of toxicity or excessive local irritation noted during this period were recorded. Ear thickness was recorded on day 1, day 3 and on day 6. The body weight of the mouse was recorded on Day 1 (prior to dosing) and Day 6.
- Compound solubility: See Table 1 in 'Any other information on materials and methods incl. tables'. Due to the physical characteristics of the test item, the 100 % (w/v) concentration was not achievable. The formulation at 50 % (w/v) using DMF as vehicle was suitable for the test.
- Irritation: no.
- Systemic toxicity: no.
- Ear thickness measurements: values were within the acceptable range, see 'Any other information on materials and methods incl. tables'.
- Erythema scores: no signs of erythema were observed. See 'Any other information on materials and methods incl. tables'.
MAIN STUDY
Groups of four mice were treated with the test item diluted at 50%, 25% and 10% in DMF, based on the results of the pre-screen tests. The mice were treated by daily application of 25 µL of the appropriate concentration of the test item to the dorsal surface of each ear for three consecutive days (Days 1, 2, 3).
- Clinical observations: All animals were observed daily on Days 1, 2, 3, 4, 5 and 6. Any signs of toxicity or signs of ill health during the test were recorded.
- Body weight: The bodyweight of each mouse was recorded on Day 1 (prior to dosing) and Day 6 (prior to termination).
- Ear thickness: On day 1 and on day 3 (before application) as well as on day 6 (after sacrifice) of each experiment, the thickness of the right ear of each animal of the vehicle control and treated groups was measured by a micrometer. Furthermore, on day 6, punch biopsies of 8 mm in diameter of the apical area of both ears were prepared and weighed in order to assess the irritation potential of the test item and the two lymph nodes per mouse were weighed.
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: Local Lymph Node Assay: BrdU - ELISA.
- Criteria used to consider a positive response: BrdU was measured by ELISA using a commercial kit (Roche Applied Science, Mannheim, Germany, Catalogue Number 11 647 229 001 – Batch No.17267000). Briefly, 100 µL of the LNC suspension was added to the wells of a flat-bottom microplate at least in triplicate. After fixation and denaturation of the LNC, anti-BrdU antibody was added to each well and allowed to react. Subsequently the anti-BrdU antibody was removed by washing and the substrate solution was then added and allowed to produce chromogen. After 5 to 30 min, 30µL of 1 M H2SO4 was added in each well, then shaken for one minute. Absorbance at 450 nm with a reference wavelength of 690 nm was then measured. The SI value was derived as specified in the guidelines. If at least one concentration of the test item results is greater than 1.6 compared to control values, that is considered a positive response.
TREATMENT PREPARATION AND ADMINISTRATION:
Groups of four mice were treated with the test item diluted at 50%, 25% and 10% in DMF, based on the results of the pre-screen tests. The mice were treated by daily application of 25 µL of the appropriate concentration of the test item to the dorsal surface of each ear for three consecutive days (Days 1, 2, 3). The test item formulation was administered using an automatic micropipette and spread over the dorsal surface of the ear using the tip of the pipette. A further group of four mice received the vehicle alone in the same manner.
On Day 5, 0.5 mL (5 mg/mouse) of BrdU (10 mg/mL) solution was injected by intra-peritoneal route. On day 6 (end of the test), the animals were anaesthetised with sodium pentobarbital and administration continued to fatal levels. The draining auricular lymph nodes from the four mice were excised.
From each mouse, a single-cell suspension of lymph node cells (LNC) excised bilaterally was prepared by gentle mechanical disaggregation through a disposable plastic pestle to crush the lymph nodes followed by passage through a #70 nylon mesh in 15 mL of DPBS (Ca2+ / Mg2+ - free) into a well of a multi-well 6. The optimised volume was based on achieving a mean absorbance of the negative control group within 0.1- 0.2. Then, BrdU was determined. - Positive control substance(s):
- hexyl cinnamic aldehyde (CAS No 101-86-0)
- Positive control results:
- The results for the last positive control (29/06/2016) were within the acceptable ranges. 5%, 10% and 25% solutions of hexyl cinnamic aldehyde in MEK generated a SI = 1.34, 1.63 and 1.95, respectively. The EC1.6 value was 9.48%.
- Key result
- Parameter:
- SI
- Value:
- 0.84
- Test group / Remarks:
- 50%
- Key result
- Parameter:
- SI
- Value:
- 1.01
- Test group / Remarks:
- 25%
- Key result
- Parameter:
- SI
- Value:
- 1.04
- Test group / Remarks:
- 10%
- Parameter:
- other: EC1.6
- Remarks on result:
- not determinable
- Cellular proliferation data / Observations:
- CELLULAR PROLIFERATION DATA: see 'Any other information on results' below.
DETAILS ON STIMULATION INDEX CALCULATION: as recommended by the guideline. No stimulation index of more than 1.6 was recorded whatever the tested concentration.
EC CALCULATION: The EC1.6 value (theoretical concentration resulting in a SI value of 1.6) was determined by linear interpolation of points on the dose-response curve, immediately above and below the 1.6-fold threshold. The equation used for calculation of EC1.6 was: EC1.6 = c + [(1.6 – d) / (b – d)] x (a – c), where a = the lowest concentration giving stimulation index > 1.6; b = the actual stimulation index caused by a; c = the highest concentration failing to produce a stimulation index of 1.6; d = the actual stimulation index caused by c. As no concentration giving stimulation index higher than 1.6, the EC1.6 cannot be determined.
CLINICAL OBSERVATIONS: No mortality and no signs of systemic toxicity were noted in the test and control animals during the test. Residual test item was noted in all animals treated at 50% between days 2 and 4. No increase in ear thickness and in ear weight was noted in animals treated at 50%, 25% and 10%. Therefore, the test item has to be considered as not excessively irritant at these concentrations.
BODY WEIGHTS: Bodyweight changes of the test animals between Day 1 and Day 6 were comparable to those observed in the corresponding control group animals over the same period. - Interpretation of results:
- GHS criteria not met
- Remarks:
- EU criteria.
- Conclusions:
- The Stimulation Indexes were 1.04, 1.01, 0.84 at concentrations of 10, 25 and 50% test item in DMF, respectively. Since all values were below 1.6, it can be concluded that the test item did not show skin sensitisation potential under the tested conditions in the LLNA assay.
- Executive summary:
The skin sensitisation potential of the test item was studied according to OECD 422B, under GLP conditions. Four groups of four female CBA/J mice received 25 µL/ear of 50, 25, and 10% (w/v) test item in DMF; and one negative control group received the vehicle (DMF). The last positive control results were used for reference. The animals were treated for 3 consecutive days (D1, D2, D3), and injected with BrdU solution on Day 5. On D6, the proliferation of lymphocytes in the draining auricular lymph nodes was determined by measurement of BrdU content in DNA of lymphocyte using an ELISA kit. The values obtained were used to calculate stimulation indices (SI). No mortality and no signs of systemic toxicity were noted in the test and control animals during the test. No increase in ear thickness and in ear weight was noted in animals treated at 10%, 25% and 50%. Therefore, the test item has to be considered as not excessively irritant at these concentrations. The Stimulation Indexes were 1.04, 1.01, 0.84 at concentrations of 10, 25 and 50% test item in DMF, respectively. In conclusion, since all values were below 1.6, under the conditions of the present assay, the test item showed no sensitisation potential.
- Endpoint:
- skin sensitisation: in vitro
- Data waiving:
- study scientifically not necessary / other information available
- Justification for data waiving:
- other:
- Reason / purpose for cross-reference:
- data waiving: supporting information
Referenceopen allclose all
Table 3. Main study: Individual clinical observation and mortality data.
Groups |
Test item |
Animal |
Day 1 |
Day 2 |
Day 3 |
Day 4 |
Day 5 |
Day 6 |
1 |
DMF |
sf 8954 |
0 |
0 |
0 |
0 |
0 |
0 |
sf 8955 |
0 |
0 |
0 |
0 |
0 |
0 |
||
sf 8956 |
0 |
0 |
0 |
0 |
0 |
0 |
||
sf 8957 |
0 |
0 |
0 |
0 |
0 |
0 |
||
2 |
10% |
sf 8974 |
0 |
0 |
0 |
0 |
0 |
0 |
Sf 8975 |
0 |
0 |
0 |
0 |
0 |
0 |
||
Sf 8976 |
0 |
0 |
0 |
0 |
0 |
0 |
||
sf 8977 |
0 |
0 |
0 |
0 |
0 |
0 |
||
3 |
25% |
sf 8979 |
0 |
0 |
0 |
0 |
0 |
0 |
sf 8980 |
0 |
0 |
0 |
0 |
0 |
0 |
||
sf 8981 |
0 |
0 |
0 |
0 |
0 |
0 |
||
Sf 8982 |
0 |
0 |
0 |
0 |
0 |
0 |
||
4 |
50% |
Sf 8984 |
0 |
0 |
0 |
0 |
0 |
0 |
Sf 8985 |
0 |
0 |
0 |
0 |
0 |
0 |
||
Sf 8986 |
0 |
0 |
0 |
0 |
0 |
0 |
||
Sf 8987 |
0 |
0 |
0 |
0 |
0 |
0 |
Table 4. Main study: Individual body weight and body weight gain.
Groups |
Test item |
Animals |
Bodyweight (g) |
Body weight gain |
|
Day 1 |
Day 6 |
||||
1 |
MEK |
N° Sf 8954 |
19.8 |
20.8 |
1 |
N° Sf 8955 |
19.6 |
21.4 |
1.8 |
||
N° Sf 8956 |
20.8 |
22.0 |
1.2 |
||
N° Sf 8957 |
20.5 |
22.0 |
1.5 |
||
MEAN |
20.2 |
21.6 |
1.4 |
||
Standard-deviation |
0.6 |
0.6 |
0.3 |
||
2 |
10% |
N° Sf 8974 |
22.1 |
23.9 |
1.8 |
N° Sf 8975 |
20.6 |
21.8 |
1.2 |
||
N° Sf 8976 |
21.8 |
22.5 |
0.7 |
||
N° Sf 8977 |
19.4 |
19.9 |
0.5 |
||
MEAN |
21.0 |
22.0 |
1.1 |
||
Standard-deviation |
1.2 |
1.7 |
0.6 |
||
3 |
25% |
N° Sf 8979 |
20.5 |
21.5 |
1 |
N° Sf 8980 |
21.4 |
22.9 |
1.5 |
||
N° Sf 8981 |
21.5 |
22.3 |
0.8 |
||
N° Sf 8982 |
19.8 |
22.0 |
2.2 |
||
MEAN |
20.8 |
22.2 |
1.4 |
||
Standard-deviation |
0.8 |
0.6 |
0.6 |
||
4 |
50% |
N° Sf 8984 |
20.8 |
21.4 |
0.6 |
N° Sf 8985 |
20.2 |
21.5 |
1.3 |
||
N° Sf 8986 |
21.7 |
22.7 |
1 |
||
N° Sf 8987 |
20.5 |
21.4 |
0.9 |
||
MEAN |
20.8 |
21.8 |
0.9 |
||
Standard-deviation |
0.6 |
0.6 |
0.3 |
DMF: dimethylformamide.
Table 5. BrdU index & Stimulation index per group and calculation of EC1.6.
Group |
Test item |
BrdU-index |
Stimulation Index |
Result |
EC1.6 value |
1 |
DMF |
0.465 |
n.a |
n.a |
n.a. |
2 |
10% |
0.484 |
1.04 ± 0.21 |
negative |
n.a. |
3 |
25% |
0.469 |
1.01 ± 0.09 |
negative |
|
4 |
50% |
0.389 |
0.84 ± 0.14 |
negative |
The BrdU labelling index is defined as:
BrdU
labelling index = (ABSem – ABS blankem) – (ABSref – ABS blankref); em =
emission wavelength; and ref = reference wavelength.
EC1.6 value: Theorical concentration resulting in a SI value of 1.6;
EC1.6 = c + [(1.6 – d) / (b – d)] x (a – c)
Legend:
a = the
lowest concentration giving stimulation index > 1.6
b = the actual stimulation index caused by a
c = the highest concentration failing to produce a stimulation index of
1.6
d = the actual stimulation index caused by c.
Table 6. Summary of results – skin irritation – 30/11/16.
Groups |
Test item |
Ear thickness |
Biopsy ear weight |
Excessive |
1 |
DMF |
-6.7 |
n.a |
No |
2 |
10% |
-4.5 |
6.1 |
No |
3 |
25% |
2.4 |
8.1 |
No |
4 |
50% |
5.1 |
3.4 |
No |
#: O.E.C.D. criteria: (%
increase in ear thickness higher than 25%, score of erythema higher than
3
Table 7. Individual Ear thickness and irritation level.
Groups |
Test item |
Animals |
Day 1 |
Day 3 |
Day 6 |
Ear thickness |
Ear thickness |
1 |
DMF |
N° Sf 8954 |
0.22 |
0.22 |
0.19 |
0.0 |
-13.6 |
N° Sf 8955 |
0.21 |
0.21 |
0.19 |
0.0 |
-9.5 |
||
N° Sf 8956 |
0.19 |
0.20 |
0.20 |
5.3 |
5.3 |
||
N° Sf 8957 |
0.22 |
0.22 |
0.20 |
0.0 |
-9.1 |
||
MEAN |
0.21 |
0.21 |
0.20 |
1.3 |
-6.7 |
||
Standard-deviation |
0.01 |
0.01 |
0.01 |
2.6 |
8.3 |
||
2 |
10% |
N° Sf 8974 |
0.22 |
0.19 |
0.21 |
-13.6 |
-4.5 |
N° Sf 8975 |
0.21 |
0.20 |
0.21 |
-4.8 |
0.0 |
||
N° Sf 8976 |
0.20 |
0.21 |
0.20 |
5.0 |
0.0 |
||
N° Sf 8977 |
0.22 |
0.22 |
0.19 |
0.0 |
-13.6 |
||
MEAN |
0.21 |
0.21 |
0.20 |
-3.3 |
-4.5 |
||
Standard-deviation |
0.01 |
0.01 |
0.01 |
7.9 |
6.4 |
||
3 |
25% |
N° Sf 8979 |
0.21 |
0.20 |
0.21 |
-4.8 |
0.0 |
N° Sf 8980 |
0.21 |
0.21 |
0.22 |
0.0 |
4.8 |
||
N° Sf 8981 |
0.21 |
0.21 |
0.21 |
0.0 |
0.0 |
||
N° Sf 8982 |
0.21 |
0.21 |
0.19 |
0.0 |
4.8 |
||
MEAN |
0.21 |
0.21 |
0.22 |
-1.2 |
2.4 |
||
Standard-deviation |
0.00 |
0.00 |
0.01 |
2.4 |
2.7 |
||
4 |
50% |
N° Sf 8984 |
0.19 |
0.21 |
0.19 |
10.5 |
0.0 |
N° Sf 8985 |
0.20 |
0.21 |
0.20 |
5.0 |
0.0 |
||
N° Sf 8986 |
0.19 |
0.20 |
0.21 |
5.3 |
10.5 |
||
N° Sf 8987 |
0.20 |
0.21 |
0.22 |
5.0 |
10.0 |
||
MEAN |
0.20 |
0.21 |
0.21 |
6.4 |
5.1 |
||
Standard-deviation |
0.01 |
0.00 |
0.01 |
2.7 |
5.9 |
Table 8. Individual Ear biopsy weight and lymph node weight.
Groups |
Test item |
Animals |
ear weight |
% of ear weight |
lymph nodes |
1 |
DMF |
N° Sf 8954 |
26.2 |
- |
4.5 |
N° Sf 8955 |
27.3 |
5.1 |
|||
N° Sf 8956 |
25.7 |
5.0 |
|||
N° Sf 8957 |
28.2 |
5.2 |
|||
MEAN |
26.9 |
5.0 |
|||
Standard-deviation |
1.1 |
0.3 |
|||
2 |
10% |
N° Sf 8974 |
29.1 |
6.1 |
5.0 |
N° Sf 8975 |
28.4 |
5.9 |
|||
N° Sf 8976 |
28.0 |
6.7 |
|||
N° Sf 8977 |
28.5 |
5.8 |
|||
MEAN |
28.5 |
5.9 |
|||
Standard-deviation |
0.5 |
0.7 |
|||
3 |
25% |
N° Sf 8979 |
28.9 |
8.1 |
6.8 |
N° Sf 8980 |
29.5 |
6.8 |
|||
N° Sf 8981 |
30.0 |
6.5 |
|||
N° Sf 8982 |
27.7 |
6.3 |
|||
MEAN |
29.0 |
6.6 |
|||
Standard-deviation |
1.0 |
0.2 |
|||
4 |
50% |
N° Sf 8984 |
28.0 |
3.4 |
4.8 |
N° Sf 8985 |
26.3 |
4.8 |
|||
N° Sf 8986 |
28.8 |
4.8 |
|||
N° Sf 8987 |
28.0 |
4.8 |
|||
MEAN |
27.8 |
4.8 |
|||
Standard-deviation |
1.1 |
0.0 |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (not sensitising)
- Additional information:
Key study. The skin sensitisation potential of the test item was studied according to OECD 422B (GLP study). Four groups of four female CBA/J mice received 25 µL/ear of 50, 25, and 10% (w/v) test item in dimethylformamide; and one negative control group received the vehicle (DMF). The last positive control results were used for reference.The animals were treated for 3 consecutive days (D1, D2, D3), and injected with BrdU solution on Day 5. On D6,the proliferation of lymphocytes in the draining auricular lymph nodes was determined by measurement of BrdU content in DNA of lymphocyte using an ELISA kit. The values obtained were used to calculate stimulation indices (SI). No increase in ear thickness and in ear weight was noted in animals treated at 50%, 25% and 10%. Therefore, the test item has to be considered as not excessively irritant at these concentrations. The Stimulation Indexes were 1.04, 1.01, 0.84 at concentrations of 10, 25 and 50% test item in DMF, respectively. All values were below 1.6. In conclusion, under the conditions of the present assay, the test item had no sensitisation potential.
Data waiving (study scientifically not necessary / other information available): data from a LLNA study according to OECD 422B was available. Therefore, an in vitro study was not deemed necessary.
Respiratory sensitisation
Endpoint conclusion
- Endpoint conclusion:
- no study available
Justification for classification or non-classification
Based on the available information (SI < 1.6), the substance is not classified for sensitising properties in accordance with CLP Regulation (EU) No. 1272/2008.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
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