Registration Dossier

Diss Factsheets

Administrative data

Description of key information

The test item at the maximum feasible concentration of 12.5 %(w/v) was shown to have no skin sensitization potential in the Local Lymph Node Assay.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
February 23 - April 28, 2017
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)
Version / remarks:
22 July 2010
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Version / remarks:
31 May, 2008
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of study:
mouse local lymph node assay (LLNA)
Specific details on test material used for the study:
Date of production: 16.06.2015
Expiration date: 16.06.2020
Species:
mouse
Strain:
CBA/Ca
Remarks:
CBA/Ca Ola Hsd Mice
Sex:
female
Details on test animals and environmental conditions:
Main Test Animals
Species and strain: CBA/Ca Ola Hsd mice
Source: TOXI-COOP ZRT. H-1103, Budapest, Cserkesz u. 90.
Hygienic level at arrival: SPF
Hygienic level during the study: Good conventional
Justification of strain: On the basis of comparative investigations in other laboratories, mice of the CBA/Ca strain were found to exhibit a more marked response than other strains. Females are used because the existing database is predominantly based on females.
Number of animals: 28 animals/main test* (4 animals/treatment group)
Sex: Female, nulliparous, non-pregnant
Age of animals: Young adult mice; 11-12 weeks old (at start of the main test)
Body weight range at starting: 17.4 – 21.5 g
The weight variation in animals involved in the study did not exceed  20 % of the mean weight.
Acclimatization time: 7 days

Husbandry
Animal health: Only healthy animals (and not showing any sign of skin lesion) were used
Housing during acclimatization period: Grouped caging in small groups
Housing during the test: Grouped caging (4 animals/cage)
Cage type: Type II. Polypropylene / polycarbonate
Bedding: Laboratory bedding
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature: 22 ± 3 °C
Relative humidity: 30 – 70 %
Housing/Enrichment: Mice were group-housed to allow social interaction, and with deep wood sawdust bedding, to allow digging and other normal rodent activities.
The same conditions were used for the dose range finding and main test animals. There were no deviations from these specifications during the experimental phase. The temperature and relative humidity were recorded daily during the acclimatization and experimental phases. Before housing the animals, the microbiological status of the room was checked.
Food and Water Supply
Animals received ssniff® Rat/Souris-Elevage E complete diet for rats and mice.

The food is periodically analysed and is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study. Copies of the relevant Certificates of Analysis are maintained in TOXI-COOP ZRT.’s archive.

Animals received tap water from watering bottles ad libitum. The drinking water is periodically analysed and is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.

Identification of Animals
The individual identification of the animals was performed by numbers on the tail. The cages were marked with identification cards, with information (at least) about study code, species, strain, sex, dose group and individual animal numbers.

Randomization
The animals were set in order of their body weight. The animals were randomly assigned to control and test groups using a randomization scheme. The randomization was checked by computer software [SPSS/PC+ (4.0.1)] according to the actual body weights verifying the homogeneity and deviations between the groups.
Vehicle:
dimethyl sulphoxide
Concentration:
test item concentrations: 12.5%, 6.25%, 3.13%, 1.56%
No. of animals per dose:
4 animals/treatment group
Details on study design:
Based on the preliminary test results, 12.5 % (w/w) was used as the maximum attainable concentration in the main test with the aim of testing the highest possible concentration and formulated according to the procedure recommended by the Sponsor. The test item was tested also at three additional lower concentrations (6.25 %, 3.13 % and 1.56 %, w/w) to evaluate a dose-response relationship and ensure validity of the test in accordance with the relevant guidelines.
Animals in the treatment groups were treated with the negative controls (vehicles), appropriate formulations of the test item or 25 % (w/v) concentration of the positive control substance. The test item was administered at four different concentrations according to the results of the dose range finding test.

In vivo Treatment
Each mouse was topically treated with 25 L of the appropriate formulations of the test item, the positive control substance or the vehicles (see Table 3) using a pipette, on the dorsal surface of each ear. After the treatments animals were returned to their cages. Each animal was dosed once a day for three consecutive days (Days 1, 2 and 3). There was no treatment on Days 4, 5 and 6.

Proliferation Assay
No animals showed symptoms of systemic toxicity or excessive skin irritation, and no technical treatment failures were observed during the test. All animals treated were processed and therefore no treatment group was excluded from the evaluation.
Injection of 3HTdR
On Day 6 each mouse was intravenously injected via the tail vein with 250 L of sterile PBS (1 x PBS, diluted from 10x concentrate) containing approximately 20 Ci# of 3H-methyl-thymidine using a hypodermic needle with 1 mL sterile syringe. Once injected, mice were left for 5 hours (± 30 minutes).
Remark: Breaking down of [methyl-3H]-Thymidine in aqueous solution (about 3 % per month) was taken into account as necessary when 3HTdR solution was prepared.

Removal and Preparation of Draining Auricular Lymph Nodes
Five hours (± 30 minutes) after intravenous injection the mice were sacrificed by cervical dislocation. The draining auricular lymph nodes were excised by making a small incision in the skin between the jaw and the sternum, pulling the skin gently back towards the ears and exposing the lymph nodes. Then the nodes were removed using forceps.
Once removed, the nodes of the mice from each test group were pooled and collected separately in a Petri dish containing a small amount (1-2 mL) of PBS to keep the nodes wet before processing.
Preparation of Single Cell Suspension of Lymph Node Cells
A single cell suspension (SCS) of lymph node cells (LNCs), pooled according to groups, was prepared and collected in disposable tubes by gentle mechanical disaggregating of the lymph nodes through a cell strainer using the plunger of a disposable syringe. The cell strainer was washed with PBS (up to 10 mL). LNCs were pelleted with a relative centrifugal force (RCF) of approximately 190 x g for 10 minutes at 4 C. After centrifugation, the supernatant was removed, leaving 1-2 mL supernatant above each pellet. The pellets were gently agitated before making up to 10 mL with PBS and re-suspending the LNCs. The washing procedure was repeated twice. This procedure was repeated for each group of pooled lymph nodes.

Determination of Incorporated 3HTdR
After the final wash, each supernatant was removed leaving a small volume (< 0.5 mL) of supernatant above each pellet. The pellets were gently agitated before suspending the LNCs in 3 mL of 5 % (w/v) trichloroacetic acid (TCA, dissolved in purified water) for precipitation of the macromolecules. After incubation with 5 % TCA at 2-8C overnight (approx. 18 hrs), each precipitate was removed by centrifugation of the samples at approximately 190 x g for 10 minutes at 4C and decanting the supernatants. Then the pellets were re-suspended in 1 mL of 5 % TCA and dispersed using an ultrasonic water bath. Samples were transferred to suitable sized scintillation vials containing 10 mL of scintillation liquid, gently mixed and stored at room temperature protected from light until measurement in the -scintillation counter. 3HTdR incorporation was measured for up to 10 minutes per sample. The -counter expressed the 3HTdR incorporation as the amount of radioactive disintegration per minute (DPM). Similarly, background 3HTdR levels were measured in two 1 mL aliquots of 5 % TCA. Instrument used for the measurement:
Name: Tri-Carb 3100TR
Liquid Scintillation Analyzer
Serial Number: 072971
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
The randomization was checked by computer software [SPSS/PC+ (4.0.1)].
Positive control results:
The positive control item (25 % (w/v) HCA in Acetone:Olive oil 4:1 (v/v) mixture, AOO) induced significant stimulation when compared to the concurrent control (SI = 6.6), thus confirming the validity of the assay.
Key result
Parameter:
EC3
Remarks on result:
not determinable
Remarks:
SI at all test concentrations below 3
Parameter:
SI
Value:
2.4
Variability:
p=0.20; r= 0.80
Test group / Remarks:
at test item concentration of 12.5%
Parameter:
SI
Value:
1
Variability:
p=0.20; r= 0.80
Test group / Remarks:
at test item concentration of 6.25%
Parameter:
SI
Value:
1.3
Variability:
p=0.20; r= 0.80
Test group / Remarks:
at test item concentration of 3.13%
Parameter:
SI
Value:
1.3
Variability:
p=0.20; r= 0.80
Test group / Remarks:
at test item concentration of 1.56%
Cellular proliferation data / Observations:
No mortality or signs of systemic toxicity were observed during the test. No significant, treatment related effect on the body weights was observed in any dose group. No signs of irritation or any other local effects were observed at the treatment site (ears) in any treatment group.
No significantly increased lymphoproliferation (indicated by an SI ≥ 3) compared to the relevant control [50 % (v/v) DMSO in water] was noted for Brown 3 at the applied test concentrations. The observed stimulation index values were 2.4, 1.0, 1.3 and 1.3 at test item concentrations of 12.5 %, 6.25 %, 3.13 % and 1.56 % (w/w), respectively. No significant dose-response relationship was observed (p = 0.20, r = 0.80; evaluated by linear regression using SI values).
According to the evaluation criteria of the relevant guidelines, the lack of a significantly increased lymphoproliferation (indicated by an SI  3) up to the maximum attainable concentration of 12.5 % (w/w) as well as the lack of a significant dose-response relationship is considered as evidence that Brown 3 is not a skin sensitizer.

Individual Body Weights of the Animals with Group Means, the Associated Error Termsand Body Weight Changes in the Main Test

Animal

Dose Group

Initial

Terminal

Body Weight

Number

Body Weight

Body Weight

Change

(g)

(g)

(%)

308

Vehicle control

20.2

21.2

5

309

 for the positive control:

18.2

18.2

0

310

AOO

20.5

19.3

-6

345

 

21.5

22.4

4

 

Mean

20.1

20.3

1

 

SD

1.4

1.9

 

311

Positive control:

18.1

17.4

-4

312

25 % HCA

20.8

20.1

-3

313

 in AOO

20.0

20.8

4

346

 

21.3

21.3

0

 

Mean

20.1

19.9

-1

 

SD

1.4

1.7

 

314

Vehicle control

19.8

19.6

-1

315

 for the test item:

20.9

20.4

-2

316

50 % (v/v) DMSO in water

17.9

18.2

2

347

 

21.0

20.9

0

 

Mean

19.9

19.8

-1

 

SD

1.4

1.2

 

317

Brown 3

17.6

17.7

1

318

12.5 %

21.1

21.8

3

319

in DMSO : water

19.3

19.1

-1

348

 

20.7

20.0

-3

 

Mean

19.7

19.7

0

 

SD

1.6

1.7

 

320

Brown 3

19.5

19.3

-1

321

6.25 %

17.6

18.1

3

322

in DMSO : water

21.1

21.4

1

349

 

20.9

20.3

-3

 

Mean

19.8

19.8

0

 

SD

1.6

1.4

 

323

Brown 3

17.4

17.4

0

324

3.13 %

19.4

19.6

1

350

in DMSO : water

21.1

19.8

-6

351

 

20.6

20.5

0

 

Mean

19.6

19.3

-2

 

SD

1.6

1.3

 

325

Brown 3

21.4

20.7

-3

326

1.56 %

19.9

19.0

-5

327

in DMSO : water

18.0

18.1

1

352

 

20.5

21.4

4

 

Mean

20.0

19.8

-1

 

SD

1.4

1.5

 

HCA =a-Hexylcinnamaldehyde                           AOO = Acetone: Olive oil 4:1 (v/v) mixture

DMSO = Dimethyl sulfoxide                                SD = Standard Deviation

Clinical Observations in the Main Test

Dose Group

Animal
Number

Days

1

2

3

4

5

6

PT

AT

PT

AT

PT

AT

Vehicle control for the positive control:
AOO

308

N

N

N

N

N

N

N

N

N

309

N

N

N

N

N

N

N

N

N

310

N

N

N

N

N

N

N

N

N

345

N

N

N

N

N

N

N

N

N

Positive control:

25 % HCA in AOO

311

N

N

N

N

N

N

N

N

N

312

N

N

N

N

N

N

N

N

N

313

N

N

N

N

N

N

N

N

N

346

N

N

N

N

N

N

N

N

N

Vehicle control for the test item:

50 % (v/v) DMSO in water

314

N

N

N

N

N

N

N

N

N

315

N

N

N

N

N

N

N

N

N

316

N

N

N

N

N

N

N

N

N

347

N

N

N

N

N

N

N

N

N

Brown 3
12.5 % in DMSO : water

317

N

N

N

N

N

N

N

N

N

318

N

N

N

N

N

N

N

N

N

319

N

N

N

N

N

N

N

N

N

348

N

N

N

N

N

N

N

N

N

Brown 3
6.25 % in DMSO : water

320

N

N

N

N

N

N

N

N

N

321

N

N

N

N

N

N

N

N

N

322

N

N

N

N

N

N

N

N

N

349

N

N

N

N

N

N

N

N

N

Brown 3
3.13 % in DMSO : water

323

N

N

N

N

N

N

N

N

N

324

N

N

N

N

N

N

N

N

N

350

N

N

N

N

N

N

N

N

N

351

N

N

N

N

N

N

N

N

N

Brown 3
1.56 % in DMSO : water

325

N

N

N

N

N

N

N

N

N

326

N

N

N

N

N

N

N

N

N

327

N

N

N

N

N

N

N

N

N

352

N

N

N

N

N

N

N

N

N

 

PT = Prior to the treatment

AT = After the treatment

HCA =a-Hexylcinnamaldehyde

AOO = Acetone: Olive oil 4:1 mixture (v/v)

DMSO = Dimethyl sulfoxide

 

N = Normal (no symptoms observed)

Erythema Scores in the Main Test

Dose Group

Animal Number

Ears

Days

1

2

3

4

5

6

PT

AT

PT

AT

PT

AT

Vehicle control for the positive control:
AOO

308

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

309

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

310

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

345

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

Positive control:
25 % HCA in AOO

311

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

312

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

313

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

346

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

Vehicle control for the test item:
50 % (v/v) DMSO in water

314

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

315

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

316

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

347

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

Brown 3
12.5 % in DMSO : water

317

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

318

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

319

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

348

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

Brown 3
6.25 % in DMSO : water

320

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

321

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

322

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

349

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

Brown 3
3.13 % in DMSO : water

323

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

324

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

350

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

351

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

Brown 3
1.56 % in DMSO : water

325

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

326

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

327

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

352

L

0

0

0

0

0

0

0

0

0

R

0

0

0

0

0

0

0

0

0

L = Left                                     R = Right                    PT = Prior to treatment                               AT = After the treatment

AOO = Acetone: Olive oil 4:1 (v/v) mixture                    HCA =a-Hexylcinnamaldehyde                  DMSO = Dimethyl sulfoxide

Visual Observations of the Lymph Nodes in the Main Test

Dose Group

Animal
Number

Appearance of Lymph Nodes

Vehicle control for the positive control:
AOO

308

N

309

N

310

N

345

N

Positive control:
25 % HCA in AOO

311

Larger than the relevant control (AOO)

312

Larger than the relevant control (AOO)

313

Larger than the relevant control (AOO)

346

Larger than the relevant control (AOO)

Vehicle control for the test item:
50 % (v/v) DMSO in water

314

N

315

N

316

N

347

N

Brown 3
12.5 % in DMSO : water

317

N

318

N

319

N

348

N

Brown 3
6.25 % in DMSO : water

320

N

321

N

322

N

349

N

Brown 3
3.13 % in DMSO : water

323

N

324

N

350

N

351

N

Brown 3
1.56 % in DMSO : water

325

N

326

N

327

N

352

N

 

AOO = Acetone: Olive oil 4:1 (v/v) mixture

HCA =a-Hexylcinnamaldehyde

DMSO = Dimethyl sulfoxide

N = Normal

DPM and Stimulation Index Values for all Groups in the Main Test

Dose Group

Measured

Group*

DPM/Mouse#

Stimulation

DPM/group

DPM

Index Values

Vehicle control for the positive control:

3007

2985.0

746.3

1.0

AOO

 

 

 

 

Positive control:

19794

19772.0

4943.0

6.6

25 % HCA in AOO

 

 

 

 

Vehicle control for the test item:

1719

1697.0

424.3

1.0

50 % (v/v) DMSO in water

 

 

 

 

Brown 3

4081

4059.0

1014.8

2.4

12.5 % in DMSO : water

 

 

 

 

Brown 3

1760

1738.0

434.5

1.0

6.25 % in DMSO : water

 

 

 

 

Brown 3

2261

2239.0

559.8

1.3

3.13 % in DMSO : water

 

 

 

 

Brown 3

2231

2209.0

552.3

1.3

1.56 % in DMSO : water

 

 

 

 

 

HCA =a-Hexylcinnamaldehyde

AOO = Acetone: Olive oil 4:1 (v/v) mixture

DMSO = Dimethyl sulfoxide

 *Group DPM = measured DPMgroup- average DPMbackground

Average DPMbackground= 22

# Number of animals/group = 4


Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of the present assay, the test substance was shown to have no skin sensitization potential in the Local Lymph Node Assay.
Executive summary:

The aim of this study was to evaluate the skin sensitization potential of the test item following dermal exposure in the Local Lymph Node Assay. Preliminary tests were performed according to the relevant guidelines to find an appropriate vehicle and the maximum applicable concentration. The maximum achievable soluble concentration was 0.05 % (w/v) in DMSO using ultrasonic dispersion and stirring. No adequate homogeneous formulation (solution or suspension) was obtained with any of the vehicles used in the solvent trial above this concentration. To find an appropriate formulation at higher concentration the test item was formulated in water resulting in a thick suspension (slurry) at a nominal concentration of 25 % (w/w) prior to dilution with an appropriate vehicle (DMSO) in a ratio of 1:1 (v/v; see section 6.1.3) according to the Sponsor’s request. The 12.5 % (w/w) formulation prepared in this way was an adequately stable and homogeneous suspension for application on the dorsum of ears of animals. No significant adverse effects (systemic toxicity or irritation) were observed in the dose range finding test up to 12.5 % (w/w) concentration. According to this the test item was examined in the main test at concentration of 12.5 % and at 6.25 %, 3.13 % or 1.56 % (w/w; prepared by serial dilution in DMSO : water 1:1 (v/v) mixture) as suspension formulations. An appropriate positive control (a- hexylcinnamaldehyde, HCA) and furthermore two negative control groups dosed with DMSO : water 1:1 (v/v) mixture (as vehicle control for the test groups) or AOO (as vehicle control for the positive control group) were employed. The positive control item (25 % (w/v) HCA in Acetone:Olive oil 4:1 (v/v) mixture, AOO) induced significant stimulation when compared to the concurrent control (SI = 6.6), thus confirming the validity of the assay. No mortality or signs of systemic toxicity were observed during the test. No significant, treatment related effect on the body weights was observed in any dose group. No signs of irritation or any other local effects were observed at the treatment site (ears) in any treatment group. No significantly increased lymphoproliferation (indicated by an SI≥ 3)compared to the relevant control [50 % (v/v) DMSO in water] was noted for the test item at the applied test concentrations. The observed stimulation index values were 2.4, 1.0, 1.3 and 1.3 at test item concentrations of 12.5 %, 6.25 %, 3.13 % and 1.56 % (w/w), respectively. No significant dose-response relationship was observed (p = 0.20, r = 0.80; evaluated by linear regression using SI values). According to the evaluation criteria of the relevant guidelines, the lack of a significantly increased lymphoproliferation

indicated by an SI3)up to the maximum attainable concentration of 12.5 % (w/w) as well as the lack of a significant dose-response relationship is considered as evidence that the test substance is not a skin sensitizer.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

The local lymph node assay was conducted to evaluate the skin sensitization potential of the test item following dermal exposure in the Local Lymph Node Assay. Preliminary tests were performed according to the relevant guidelines to find an appropriate vehicle and the maximum applicable concentration. The maximum achievable soluble concentration was 0.05 % (w/v) in DMSO using ultrasonic dispersion and stirring. To find an appropriate formulation at higher concentration the test item was formulated in water resulting in a thick suspension (slurry) at a nominal concentration of 25 % (w/w) prior to dilution with an appropriate vehicle (DMSO) in a ratio of 1:1 (v/v; see section 6.1.3) according to the Sponsor’s request. The 12.5 % (w/w) formulation prepared in this way was an adequately stable and homogeneous suspension for application on the dorsum of ears of animals. No significant adverse effects (systemic toxicity or irritation) were observed in the dose range finding test up to 12.5 % (w/w) concentration. According to this the test item was examined in the main test at concentration of 12.5 % and at 6.25 %, 3.13 % or 1.56 % (w/w; prepared by serial dilution in DMSO : water 1:1 (v/v) mixture) as suspension formulations. An appropriate positive control (a- hexylcinnamaldehyde, HCA) and furthermore two negative control groups dosed with DMSO : water 1:1 (v/v) mixture (as vehicle control for the test groups) or AOO (as vehicle control for the positive control group) were employed. The positive control item (25 % (w/v) HCA in Acetone:Olive oil 4:1 (v/v) mixture, AOO) induced significant stimulation when compared to the concurrent control (SI = 6.6), thus confirming the validity of the assay. No mortality or signs of systemic toxicity were observed during the test. No significant treatment related effect on the body weights was observed in any dose group. No signs of irritation or any other local effects were observed at the treatment site (ears) in any treatment group. No significantly increased lymphoproliferation (indicated by an SI≥ 3)compared to the relevant control was noted for the test item at the applied test concentrations. The observed stimulation index values were 2.4, 1.0, 1.3 and 1.3 at test item concentrations of 12.5 %, 6.25 %, 3.13 % and 1.56 % (w/w), respectively. No significant dose-response relationship was observed (p = 0.20, r = 0.80; evaluated by linear regression using SI values). According to the evaluation criteria of the relevant guidelines, the lack of a significantly increased lymphoproliferation indicated by an SI3) up to the maximum attainable concentration of 12.5 % (w/w) as well as the lack of a significant dose-response relationship is considered as evidence that the test substance is not a skin sensitizer.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

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 sensitisation, the test item is not classified according to Regulation (EC) No 1272/2008 (CLP), as amended for the twelfth time in Regulation (EU) No 2019/521.


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