Registration Dossier
Registration Dossier
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 220-076-0 | CAS number: 2623-23-6
Endpoint summary
Administrative data
Description of key information
Based on the available in vivo test (OECD 429) the test item is not considered as skin sensitizer.
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 2012-04-11 to 2012-06-12
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of study:
- mouse local lymph node assay (LLNA)
- Species:
- mouse
- Strain:
- CBA
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Age at study initiation: 8-11 weeks
- Weight at study initiation: 19.7-23.0 g
- Housing: group housing
- Diet (e.g. ad libitum): Teklad Global Rodent 2914C – Rezeptur 3255 GLP Batch 73/11 (Provimi Kliba AG, 4303 Kaiseraugst/Switzerland), available ad libitum.
- Water (e.g. ad libitum): Community tap water from Itingen/Switzerland, available ad libitum.
- Acclimation period: 6 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.5-20.4
- Humidity (%): 37.4-62.2
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12 - Vehicle:
- acetone/olive oil (4:1 v/v)
- Concentration:
- 25, 50, 100% (w/v) in vehicle
- No. of animals per dose:
- 4
- Details on study design:
- RANGE FINDING TESTS:
- Compound solubility: The highest test item concentration which can be technically used was a 75% (w/w) solution in acetone:olive oil (4:1 v/v). As the test item is a well pipettable liquid, it was also tested neat (100%).
- Irritation: At the tested 50 and 100% concentrations, the animals did not show any signs of irritation.
- Lymph node proliferation response: is expressed as the number of radioactive disintegrations per minute per lymph node (dpm/lymph node) and as the ratio of 3HTdR incorporated into lymph node cells of the test groups relative to that recorded for the control group (Stimulation Index S.I.). Before dpm/lymph node values were determined, the mean scintillation-background dpm was subtracted from test and control raw data.
MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: Topical application / Administration of 3HTdR / Determination of incorporated 3HTdR
- Criteria used to consider a positive response: A test item is regarded as a sensitizer in the LLNA if the following criteria are fulfilled:
- First, the exposure to at least one concentration of the test item resulted in an incorporation of 3HTdR at least 3-fold or greater than that recorded in control mice, as indicated by the S.I.
- Second, the data are compatible with a conventional dose response, although allowance must be made (especially at high topical concentrations) for either local toxicity or immunological suppression.
TREATMENT PREPARATION AND ADMINISTRATION: Test item preparation was made freshly before each dosing occasion, no more than 4 hours prior to application to the ears. - Positive control substance(s):
- hexyl cinnamic aldehyde (CAS No 101-86-0)
- Statistics:
- For the body weight mean values and standard deviations were calculated.
- Key result
- Parameter:
- SI
- Value:
- 1.3
- Test group / Remarks:
- 25% test substance
- Key result
- Parameter:
- SI
- Value:
- 1.9
- Test group / Remarks:
- 50% test substance
- Key result
- Parameter:
- SI
- Value:
- 0.7
- Test group / Remarks:
- 100% test substance
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- The test item menthyl acetate (racemic) was not a skin sensitiser in this assay.
- Executive summary:
In order to study the possible contact allergenic potential of Menthyl acetate racemic, three groups of four female mice each were treated daily with the test item at concentrations of 25%, 50% (w/v) in acetone:olive oil (4:1 v/v) and 100% by topical application to the dorsum of each ear lobe for three consecutive days. A control group of four female mice was treated with the vehicle acetone:olive oil (4:1 v/v) only.
Five days after the first topical application the mice were injected intravenously into a tail vein with radio-labelled thymidine (3H-methyl thymidine, 3HTdR). Approximately five hours after intravenous injection, the mice were sacrificed, the draining auricular lymph nodes were excised and pooled per group. Single cell suspensions of lymph node cells were prepared from pooled lymph nodes which were subsequently washed and incubated with trichloroacetic acid overnight. The proliferative capacity of pooled lymph node cells was determined by the incorporation of 3HTdR measured in a β-scintillation counter.
All treated animals survived the scheduled study period. Neither clinical signs on the ears of the animals nor systemic findings were observed during the study period.
The results obtained [Stimulation Index (S.I.)] are reported in the following table. The estimated concentration of test item required to produce a S.I. of 3 is referred to as the EC3 value.
Test item concentration % (w/v)
SI
Group 2
25
1.3
Group 3
50
1.9
Group 4
100
0.7
EC3: -- %
A dose-response relationship was not observed. A calculation of the EC3 value was not performed because no test concentrations produced a S.I. of 3 or higher.
In conclusion, the test item Menthyl acetate (racemic) was not a skin sensitizer in this assay.
- Endpoint:
- skin sensitisation: in vivo (LLNA)
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
Please refer to the attached read across justification in section 13. - Reason / purpose for cross-reference:
- read-across source
- Key result
- Parameter:
- SI
- Value:
- 1.3
- Test group / Remarks:
- 25% test substance
- Key result
- Parameter:
- SI
- Value:
- 1.9
- Test group / Remarks:
- 50% test substance
- Key result
- Parameter:
- SI
- Value:
- 0.7
- Test group / Remarks:
- 100% test substance
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- L-menthyl acetate was estimated to be not a skin sensitiser.
- Executive summary:
One study is available from the structural analogue menthyl acaetate CAS 89 -48 -5. It is concluded that L-menthyl acetate is not a skin sensitizer as found from the study with the structural analogue. As explained in the justification for type of information, the differences in molecular structure between the target and the source are unlikely to lead to differences in the skin sensitisation potential that are higher than the typical experimental error of the test method.
Referenceopen allclose all
The following results were obtained for menthyl acetate racemic using acetone:olive oil (4:1 v/v) as vehicle:
Test item concentration % (w/v) |
Groups |
Measurement dpm |
Calculation |
Result SI |
||
dpm-BG |
Number of lymph nodes isolated |
dpm / lymph nodea) |
||||
-- |
BG I |
1 |
-- |
-- |
-- |
-- |
-- |
BG II |
1 |
-- |
-- |
-- |
-- |
-- |
Group 1 |
1327 |
1326 |
8 |
166 |
-- |
25 |
Group 2 |
1735 |
1734 |
8 |
217 |
1.3 |
50 |
Group 3 |
2549 |
2548 |
8 |
319 |
1.9 |
100 |
Group4 |
983 |
982 |
8 |
123 |
0.7 |
BG = Background (1 mL 5% trichloroacetic acid) in duplicates
SI = Stimulation Index (Ratio = dpm/lymph node test item treated group / dpm/lymph node Group 1)
a) = Since the lymph nodes of the animals of a dose group were pooled, dmp/lymph node was determined by dividing the measured value by the number of lymph nodes pooled.
The EC3 value could not be calculated, since SI values were below 3.
The following results were obtained for menthyl acetate racemic using acetone:olive oil (4:1 v/v) as vehicle:
Test item concentration % (w/v) |
Groups |
Measurement dpm |
Calculation |
Result SI |
||
dpm-BG |
Number of lymph nodes isolated |
dpm / lymph nodea) |
||||
-- |
BG I |
1 |
-- |
-- |
-- |
-- |
-- |
BG II |
1 |
-- |
-- |
-- |
-- |
-- |
Group 1 |
1327 |
1326 |
8 |
166 |
-- |
25 |
Group 2 |
1735 |
1734 |
8 |
217 |
1.3 |
50 |
Group 3 |
2549 |
2548 |
8 |
319 |
1.9 |
100 |
Group4 |
983 |
982 |
8 |
123 |
0.7 |
BG = Background (1 mL 5 % trichloroacetic acid) in duplicates
SI = Stimulation Index (Ratio = dpm/lymph node test item treated group / dpm/lymph node Group 1)
a) = Since the lymph nodes of the animals of a dose group were pooled, dmp/lymph node was determined by dividing the measured value by the number of lymph nodes pooled.
The EC3 value could not be calculated, since SI values were below 3.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (not sensitising)
- Additional information:
In order to study the possible contact allergenic potential of Menthyl acetate racemic, three groups of four female mice each were treated daily with the test item at concentrations of 25%, 50% (w/v) in acetone:olive oil (4:1 v/v) and 100% by topical application to the dorsum of each ear lobe for three consecutive days. A control group of four female mice was treated with the vehicle acetone:olive oil (4:1 v/v) only.
Five days after the first topical application the mice were injected intravenously into a tail vein with radio-labelled thymidine (3H-methyl thymidine, 3HTdR). Approximately five hours after intravenous injection, the mice were sacrificed, the draining auricular lymph nodes were excised and pooled per group. Single cell suspensions of lymph node cells were prepared from pooled lymph nodes which were subsequently washed and incubated with trichloroacetic acid overnight. The proliferative capacity of pooled lymph node cells was determined by the incorporation of 3HTdR measured in a β-scintillation counter.
All treated animals survived the scheduled study period. Neither clinical signs on the ears of the animals nor systemic findings were observed during the study period.
The results obtained [Stimulation Index (S.I.)] are reported in the following table. The estimated concentration of test item required to produce a S.I. of 3 is referred to as the EC3 value.
Test item concentration % (w/v)
SI
Group 2
25
1.3
Group 3
50
1.9
Group 4
100
0.7
EC3: -- %
A dose-response relationship was not observed. A calculation of the EC3 value was not performed because no test concentrations produced a S.I. of 3 or higher.
In conclusion, the test item Menthyl acetate (racemic) was not a skin sensitizer in this assay.
Justification for classification or non-classification
Classification,
Labelling, and Packaging Regulation (EC) No 1272/2008
The
available experimental test data are reliable and suitable for
classification purposes under Regulation (EC) No 1272/2008. Based on
available data on skin sensitizationthe
test item is not classifiedaccording
to Regulation (EC) No 1272/2008 (CLP), as amended for the eighth time in
Regulation (EU) No 2016/918.
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.
