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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Description of key information

Skin sensitisation (based on read across information from OECDTG429 study): not sensitising

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
2017
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Results derived from a valid read across, with adequate and reliable documentation/justification.
Justification for type of information:
The read across justification is presented in the Endpoint summary of Sensitisation. The corresponding documentation file is also attached there.
Reason / purpose for cross-reference:
read-across source
Key result
Parameter:
EC3
Remarks:
%
Remarks on result:
other: EC3 was not reached
Parameter:
SI
Value:
0.8
Remarks on result:
other: 5% test group
Parameter:
SI
Value:
0.7
Remarks on result:
other: 10% test group
Parameter:
SI
Value:
0.9
Remarks on result:
other: 25% test group
Interpretation of results:
other: Not skin sensitising.
Remarks:
According to Regulation (EC) No 1272/2008 and its amendments.
Conclusions:
The test substance was considered not to be a skin sensitiser, based on the results of the source substance.
Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
15 September 2016 - 19 October 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Information used for read across to Veilex #3.
Qualifier:
according to guideline
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Version / remarks:
July 2010
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Version / remarks:
May 2008
Deviations:
no
GLP compliance:
yes
Type of study:
mouse local lymph node assay (LLNA)
Species:
mouse
Strain:
CBA:J
Sex:
female
Details on test animals and environmental conditions:
Female CBA/J strain mice were supplied by Janvier, Le Genest-Saint-Isle, France. On receipt the animals were randomly allocated to cages. The animals were nulliparous and non pregnant and the acclimatisation period was at least five days. At the start of the study the animals were in the weight range of 18.2 to 25.2 g, and were approximately ten weeks old.

Animals were group housed in labeled Makrolon cages containing sterilised sawdust as bedding material. Paper and shelters were supplied as cage-enrichment. Free access to tap water and food (pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany)) was allowed throughout the study.
The temperature and relative humidity were controlled to remain within target ranges of 18 to 24°C a nd 40 to 70 %, respectively.

The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
Vehicle:
acetone/olive oil (4:1 v/v)
Concentration:
Undiluted test item or the test item at concentrations of 5%, 10% or 25% v/v in vehicle.
No. of animals per dose:
Groups of five mice were treated per dose.
Details on study design:
Preliminary Screening Test:
A pre-screen test was conducted in order to select the highest test item concentration to be used in the main study. In principle, this highest concentration should cause no systemic toxicity, may give well-defined irritation as the most pronounced response (maximum grade 2) and/or an increase in ear thickness < 25%) and/or is the highest possible concentration that can be technically applied.
Initially, two test item concentrations were tested; a 50% and 100% concentration. The highest concentration was the maximum concentration as required in the test guidelines.
Based on the results of the initially treated animals, four additional animals were treated in a similar manner with two lower concentrations (10% and 25%) at a later stage.
The test system, procedures and techniques were identical to those used in the main study except that the animals were approximately 10-11 weeks (at initiation of treatment) and that the assessment of lymph node proliferation and necropsy were not performed. Two young adult animals per concentration were selected. Each animal was treated with one concentration on three consecutive days. Animals were group housed in labeled Makrolon cages (MII type, height 14 cm). Ear thickness measurements were conducted using a digital thickness gauge (Kroeplin C110T-K) prior to dosing on Days 1 and 3, and on Day 6.
Animals were sacrificed after the final observation.

Main Test
Test Item Administration
Groups of five mice were treated with the undiluted test item or the test item at concentrations of 5%, 10 % or 25 % v/v in acetone/olive oil 4:1. The vehicle was selected on the basis of the test item data provided by the Sponsor and trial preparation results performed at Charles River Den Bosch. The vehicle was chosen from the vehicles specified in the test guideline: Acetone/Olive oil (4:1 v/v), N,N-dimethylformamide, methylethylketone, propylene glycol, dimethylsulfoxide and 1% Pluronic© L92 in Elix water (in case an aqueous vehicle is suitable). There was no information available regarding the solubility or stability in vehicle. No analysis was conducted to determine the homogeneity, concentration or stability of the test item formulation. This is an exception with regard to GLP and has been reflected in the GLP compliance statement. The preliminary screening test suggested that the test item would not produce systemic toxicity or excessive local irritation at the highest suitable concentration. The dorsal surface of both ears was topically treated (25 μL/ear) with the test item, at approximately the same time on each day. The concentrations were stirred with a magnetic stirrer immediately prior to dosing. A further group of five mice received the vehicle alone in the same manner.

The positive control animals were similarly treated to the test animals except that 25 μL of the positive control item, α Hexylcinnamaldehyde, techical grade, at a concentration of 5, 10 and 25 % v/v in acetone/olive oil 4:1 was applied to the dorsal surface of each ear. The positive control group served as common control with another study according to the summary of positive control data for the local lymph node assay.

Induction - Days 1, 2 and 3
The dorsal surface of both ears was topically treated (25 μL/ear) with the test substance, at approximately the same time on each day. The concentrations were stirred with a magnetic stirrer immediately prior to dosing.

3H-Methyl Thymidine Administration
Five days following the first topical application of the test item, each animal was injected via the tail vein with 0.25 mL of sterile phosphate buffered saline (PBS) (Merck, Darmstadt, Germany) containing 20 μCi of 3H-methyl thymidine (PerkinElmer Life and Analytical Sciences, Boston, MA, US).

Terminal Procedures - Day 6
Termination: After five hours, all animals were killed by intraperitoneal injection (0.2 mL/animal) of Euthasol® 20% (AST Farma BV, Oudewater, The Netherlands). The draining (auricular) lymph node of each ear was excised. The relative size of the nodes (as compared to normal) was estimated by visual examination and abnormalities of the nodes and surrounding area were recorded. The nodes were pooled for each animal in approximately 3 mL PBS.

Preparation of Single Cell Suspension: Following excision of the nodes, a single cell suspension of lymph node cells (LNC) was prepared in PBS by gentle separation through stainless steel gauze
(diameter: 125 μm). LNC were washed twice with an excess of PBS by centrifugation at 200g for 10 minutes at 4ºC. To precipitate the DNA, the LNC were exposed to 5% trichloroacetic acid (TCA) (Merck, Darmstadt, Germany) and then stored in the refrigerator until the next day.

Determination of 3HTdR Incorporation: Precipitates were recovered by centrifugation, resuspended in 1 mL TCA and transferred to 10 mL of Ultima Gold cocktail (PerkinElmer Life and Analytical Sciences, Boston, MA, US) as the scintillation fluid. Radioactivity measurements were performed using a Packard scintillation counter (2800TR). Counting time was to a statistical precision of ± 0.2% or a maximum of 5 minutes whichever came first. The scintillation counter was programmed to automatically subtract back ground and convert Counts Per Minute (CPM) to Disintegrations Per Minute(DPM).

Observations
Mortality/Viability: Twice daily.
Clinical Observations: Once daily on Days 1-6 (on Days 1-3 between 3 and 4 hours after dosing). Any signs of toxicity or signs of ill health during the test were recorded.
Bodyweights: The bodyweight of each mouse was recorded on Day 1 (prior to dosing) and on Day 6 (prior to termination).
Irritation: Once daily on Days 1-6 (on Days 1-3 within 1 hour after dosing) according to the numerical scoring system (see any other information on materials and methods incl. tables). In addition, a description of all other (local) effects was recorded.
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
Not performed.
Positive control results:
The positive control item, α-Hexylcinnamaldehyde, techical grade, gave a Stimulation Index of greater than 3 when tested at a concentration of 25 % v/v in acetone/olive oil 4:1.
Key result
Parameter:
EC3
Remarks:
%
Remarks on result:
other: EC3 was not reached
Parameter:
SI
Value:
0.8
Remarks on result:
other: 5% test group
Parameter:
SI
Value:
0.7
Remarks on result:
other: 10% test group
Parameter:
SI
Value:
0.9
Remarks on result:
other: 25% test group
Cellular proliferation data / Observations:
CELLULAR PROLIFERATION DATA
DETAILS ON STIMULATION INDEX CALCULATION
The SI values calculated for the test item concentrations 5, 10 and 25% were 0.8, 0.7 and 0.9, respectively.
EC3 CALCULATION
There was no indication that the test item elicits a SI ≥ 3 when tested up to 25%, Veilex 4 was not considered to be a skin sensitizer. It was established that the EC3 value (the estimated test item concentration that will give a SI =3) (if any) exceeds 25%.
CLINICAL OBSERVATIONS/BODY WEIGHTS:
No mortality occurred and no clinical signs of systemic toxicity were observed in the animals of the main study. Body weights and body weight gain of experimental animals remained in the same range as controls over the study period.

Pre-screen Test:

At a concentration of 50%, hunched posture was noted on Day 3 and slight erythema was noted on Days 2 and 3. At a concentration of 100%, slight erythema was noted on Day 3 and scaliness was noted between Days 2 and 6. At concentrations of 25% and 10%, no signs of systemic toxicity or erythema were noted. Variations in ear thickness during the observation period were less than 25% from Day 1 pre-dose values for all animals.

Based on these results, the highest test item concentration selected for the main study was a 25% concentration.

Interpretation of results:
other: Not sensitising.
Remarks:
According to Regulation (EC) No 1272/2008 and its amendments.
Conclusions:
The SI values calculated for the test item concentrations 5, 10 and 25% were 0.8, 0.7 and 0.9, respectively. These results show that the test substance does not elicit a SI ≥ 3. An EC3 is derived of >25%. A NOEC of >25% is derived. The test substance was considered not to be a sensitiser under the conditions of the test.
Executive summary:

The skin sensitisation potential of the test substance has been tested according to OECD TG 429: Local Lymph Node Assay" method and GLP principles. The test substance was tested at 5, 10 and 25%.

No higher dose was tested in the main study, as in the preliminary study systemic effects were observed- at the concentration of 50% hunched posture was noted (dose at 100% is ca 2400 mg/kg bw (50 ul = 50 mg/0.021 kg mouse). At the concentration of 100%, scaliness was noted. The substance showed SI values of 0.8, 0.7 and 0.9, respectively. Reliable negative and positive controls were included. The test substance did not elicit an SI ≥ 3 when tested up to and including the maximum concentration of 25% and therefore the substance was not considered to be a skin sensitizer.

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
10 February 1976
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
GLP compliance:
no
Type of study:
patch test
Justification for non-LLNA method:
A Human Repeated Insult Patch Test was available for the substance. Since this study assesses the potential for sensitisation of human skin directly, it is a reliable data source.
Species:
other: human
Sex:
not specified
Details on test animals and environmental conditions:
A group of 50 individuals who qualified was selected from the local population. All of these individuals volunteered to participate in this evaluation. The criteria for qualifying were:
1. General well-being
2. Absence of any visible skin disease which might be confused with skin reactions from the test material.
3. Willingness to cooperate.
4. Dependability and intelligence in following directions.
5. Reading, understanding and signing an informed-consent statement (in the case of minors, parental consent was obtained).
Route:
epicutaneous, occlusive
Vehicle:
unchanged (no vehicle)
Concentration / amount:
100%
Day(s)/duration:
Exposure duration: 24 hours, followed by 24 or 48 hour rest period, repeated 15 times.
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
unchanged (no vehicle)
Concentration / amount:
100%
Day(s)/duration:
24 hours
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
50 human volunteers
Details on study design:
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 15
- Exposure period: 24 hours
- Test groups: 50 individuals
- Control group: no
- Site: a patch of 3 by 3 cm on the upper arm
- Frequency of applications: Mondays, Wednesdays and Fridays (alternate-day applications)
- Concentrations: 100% of the substance applied as received

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Days after induction exposure: 14
- Exposure period: 24 hours
- Test groups: 50 individuals
- Control group: no
- Site: a patch of 3 by 3 cm on the upper arm (same site as in the Induction Exposure)
- Concentrations: 100% of the substance applied as received
- Evaluation (hr after challenge): 0, 24 and 48 hours
Positive control substance(s):
no
Key result
Reading:
other: Challenge
Hours after challenge:
0
Group:
test chemical
Dose level:
100%
No. with + reactions:
0
Total no. in group:
50
Key result
Reading:
other: Challenge
Hours after challenge:
24
Group:
test chemical
Dose level:
100%
No. with + reactions:
0
Total no. in group:
50
Key result
Reading:
other: Challenge
Hours after challenge:
48
Group:
test chemical
Dose level:
100%
No. with + reactions:
0
Total no. in group:
50

Table 1 Total number of reactions

No. of application

Number of reactions, indicated in grades

0

1+

2+

3+

4+

1

50

0

0

0

0

2

49

1

0

0

0

3

45

5

0

0

0

4

35

15

0

0

0

5

50

0

0

0

0

6

25

25

0

0

0

7

39

10

1

0

0

8

29

19

2

0

0

9

47

3

0

0

0

10

45

5

0

0

0

11

43

5

2

0

0

12

44

6

0

0

0

13

42

6

2

0

0

14

46

4

0

0

0

15

46

4

0

0

0

Challenge

50

0

0

0

0

Scoring criteria:
0 = no reactions
1+ = slight erythema
2+ = marked erythema
3+ = marked erythema, edema, with or without a few vesicles
4+ = marked erythema, edema, with vesicles and oozing

Interpretation of results:
other: Not a skin sensitiser.
Remarks:
According to Regulation (EC) No 1272/2008 and its amendments.
Conclusions:
Under the conditions of this HRIPT, the substance did not produce skin sensitising reactions after the challenge.
Executive summary:

The substance was assessed for its skin sensitising potential. A Human Repeated Insult Patch Test was performed with 50 human volunteers which were exposed to 100% of the substance on a patch of 3 by 3 cm on the upper arm. The skin was exposed to the substance in an occlusive manner for 24 hours, followed by a 24 or 48 hour rest period. This cycle was repeated in the same manner for 15 times. After the fifteenth application, the participants had a 14 day rest period, followed by a challenge test. In the challenge, 100% test material was applied to the same sites under occlusion for 24 hours. After the 24 hours exposure, the skin sites were examined immediately, after 24 hours and after 48 hours. No skin reactions were observed after the challenge.

Endpoint:
skin sensitisation: in vitro
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

For skin sensitisation a supporting study on Veilex#3 is available, which is presented first. Secondly, the summary of the key study from the analogue Veilex #4 is included and finally the read-across rationale is presented.

Veilex#3 Supporting HRIPT information

The substance was assessed for its skin sensitising potential. A Human Repeated Insult Patch Test was performed with 50 human volunteers which were exposed to 100% of the substance on a patch of 3 by 3 cm on the upper arm. The skin was exposed to the substance in an occlusive manner for 24 hours, followed by a 24 or 48 hour rest period. This cycle was repeated in the same manner for 15 times. After the fifteenth application, the participants had a 14 day rest period, followed by a challenge test. In the challenge, 100% test material was applied to the same sites under occlusion for 24 hours. After the 24 hours exposure, the skin sites were examined immediately, after 24 hours and after 48 hours. No skin reactions were observed after the challenge.

Veilex#4: LLNA test information

The skin sensitisation potential of the test substance has been tested according to OECD TG 429: Local Lymph Node Assay" method and GLP principles. The test substance was tested at 5, 10 and 25%.

No higher dose was tested in the main study, as in the preliminary study systemic effects were observed- at the concentration of 50% hunched posture was noted (dose at 100% is ca 2400 mg/kg bw (50 ul = 50 mg/0.021 kg mouse). At the concentration of 100%, scaliness was noted. The substance showed SI values of 0.8, 0.7 and 0.9, respectively. Reliable negative and positive controls were included. The test substance did not elicit an SI ≥ 3 when tested up to and including the maximum concentration of 25% and therefore the substance was not considered to be a skin sensitizer.

Read across justification: 

The skin sensitization of Veilex #3 (1-cyclohexylethanol; CAS 1193-81-3) using read across from Veilex #4 (4-isopropylcyclohexyl propionate; CAS 63449-95-6)

 

Introduction and hypothesis for the analogue approach

Veilex #3 consists of a cyclohexyl backbone with attached an ethanol group.For Veilex #3 limited skin sensitization data are available, which are not sufficient to fulfill the requirement for this endpoint. Therefore, additional information is used in accordance with Article 13 of REACH, where it is said that lacking information should be generated whenever possible by means other than vertebrate animal tests, i.e. applying alternative methods such as in vitro tests, QSARs, grouping and read-across. For assessing the sensitizing potential of Veilex #3 the analogue approach is selected because for a closely related analogue, Veilex #4, sensitization information is available which can be used for read across.

Hypothesis: Veilex #3 has similar sensitization potential compared to Veilex #4.

Available information:For the target substance, Veilex #3, a HRIPT was conducted with the neat test material (100%, Klimisch 2) using 50 human volunteers.No skin sensitization was observed after challenge. This HRIPT study is not considered to be sufficient for classification and labelling and therefore the study is only used as supporting evidence.

For Veilex #4 an LLNA is available, which was performed according to OECD TG 429 and GLP principles (Klimisch 1). The test substance was tested at 5, 10 and 25%. No higher dose was tested in the main study, as in the preliminary study systemic effects were observed; at the concentration of 50% hunched posture was noted. At the concentration of 100%, skin irritation, such as scaliness, was noted. The substance showed SI values of 0.8, 0.7 and 0.9, respectively. Reliable negative and positive controls were included. The test substance did not elicit an SI ≥ 3 when tested up to and including the maximum concentration of 25% and therefore the substance was not considered to be a skin sensitizer.

Target chemical and source chemicals

Chemical structures of Veilex #3 and Veilex #4 are shown in the data matrix, including physico-chemical properties and toxicological information, thought relevant for sensitization.

Purity / Impurities

Veilex #3 is a mono-constituent presenting a purity of 95-100%.Veilex #4 is a multi-constituent substance, consisting of two main constituents which are isomers of each other and which are not expected to differ in skin sensitizing properties.

Analogue approach justification

According to Annex XI 1.5 read across can be used to replace testing when the similarity can be based on a common backbone and a common functional group.

Analogue selection: Two analogues were considered for which skin sensitisation information was available: Veilex#4 and Herbac. Veilex#4 is the propylester of Veilex#3, the alcohol. Herbac (Cas no.25304-14-7) is has a ketone group instead of an alcohol at the same position as Veilex#3. The LLNA of Veilex#4 is considered the key study because the LLNA is already for decades the approved method for skin sensitisation.

Structural similarities and differences:Veilex #3 and Veilex #4 have a common cyclohexyl backbone. For Veilex #3 the cyclohexane is substituted with an isopropyl alcohol group: the functional group being an alcohol. InVeilex #4 the cyclohexane is substituted on C1 with a hydroxyl group esterified with propanoate and an isopropyl group on C4: the functional group being an ester. 

Toxico-kinetic, Absorption:Veilex #3 and Veilex #4 indicate dermal absorption based on molecular weight (128.21 and 198.3, respectively) and both substances being liquid, despite difference in log Kow and water solubility.Metabolism:In view of Veilex #4 being an ester, it will be cleaved by carboxylesterases in the skin producing an alcohol. The alcohol Veilex #3 and the formed alcohol of Veilex #4 are considered to have similar skin penetrating properties. 

Skin sensitisation, Reactivity:No protein binding alerts are presented for skin sensitisation in the OECD toolbox for both Veilex #3 and Veilex #4. Veilex#3 and Veilex#4 differ in their functional group: alcohol and ester, the latter being somewhat more reactive and because Veilex#3 is less reactive skin sensitisation is not expected either.

Uncertainty of the prediction:Beside the information on Veilex#4, the other analogue Herbac (EC no. 944-298-9, Cas no. 25304-14-7, a dimethyl cyclohexyl ethanone containing a keton as functional group) further supports the absence of skin sensitisation because this substance is negative in the vitro testing battery, presenting negative results in the DPRA, KeratinoSens and h-CLAT, which results in absence of skin sensitisation.The absence of skin sensitisation in the HRIPT, tested at 100% also presents absence of skin sensitisation. There are no remaining uncertainties other than those already addressed.

Data MatrixThe relevant information on physico-chemical properties and toxicological characteristics are presented in the Data matrix below.

Conclusions on skin sensitisation

For Veilex#3 only HRIPT information is available resulting in absence of skin sensitisation. Additional information on analogues is presented to support this absence.When using read across the result derived should be applicable for C&L and/or risk assessment and be presented with adequate and reliable documentation. This documentation is presented in the current document.This absence of skin sensitization is further confirmed with a well conducted LLNA (OECD TG 429) study on the analogue Veilex #4 and negative in vitro skin sensitisation for the analogue Herbac, which both can be used for read across to Veilex #3. Taken all data together, Veilex #3 was concluded not to have skin sensitization properties.

Final conclusion: Veilex #3 is considered to be not skin sensitizing.

Data matrix for the read across to Veilex #3 from Veilex #4

Common names

Veilex #3

Veilex #4

 

Target

Source

 

1-cyclohexylethanol

4-isopropylcyclohexyl propionate

 

 

Chemical structures

CAS no

1193-81-3

63449-95-6

Einecs

214-780-7

264-158-4

REACH registered

Registered as an intermediate and for 2018

Registered

Empirical formula

C8H16O

C12H22O2

Molecular weight

128.21

198.3

Physico-chemical data

 

 

Physical state

Liquid

Liquid

Freezing point,oC

< -20

< -20

Boiling point,oC

195.8

250.1

Vapour pressure, Pa

36.8 (at 24°C)

5.35 (at 24°C)

Water solubility, mg/L

5163.2 (at 24°C)

13.3 (at 24°C)

Log Kow

1.6 (at 25°C)

5.1 (at 25°C)

Human health endpoints

 

 

Skin sensitisation

Derived from read across:

Not sensitizing; supported with HRIPT information

 

Not sensitising (LLNA, OECD TG 429)

 

Justification for classification or non-classification

Based on these results, the substance is not classified as a skin sensitiser according to Regulation (EC) 1272/2008 and its amendments.