sodium;2-chloroprop-2-enoate;hydrate

Endpoint summary

Description of key information

Based on a weight of evidence evaluation of the available data from three in vitro and in chemico assays covering key event 1 to 3, the test item is considered to be skin sensitising:

- DPRA: positive
- Keratinosens: positive

- h-Clat: negative

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Reference 1

Endpoint:

skin sensitisation: in chemico

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2018-04-13 to 2019-05-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 442C (In Chemico Skin Sensitisation: Direct Peptide Reactivity Assay (DPRA))

GLP compliance:

yes (incl. QA statement)

Type of study:

direct peptide reactivity assay (DPRA)

Details on the study design:

The reactivity of a test chemical and synthetic cysteine or lysine containing peptides was evaluated by combining the test chemical with a solution of the peptide (reaction samples) and monitoring the remaining concentration of the peptide following 24 ± 2 hours of interaction time at room temperature (25 ± 2.5 °C). The peptide is a custom material containing phenylalanine to aid in detection and either cysteine or lysine as the reactive centre. Relative concentrations of the peptide following the 24 hour reaction time were determined by HPLC with gradient elution and UV detection at 220 nm. Samples were prepared and analysed in triplicates in batches to keep the total HPLC analysis time less than 30 hours.

Positive control results:

Peptide depletion resulted from the positive control cinnamaldehyde was 70.60 % with cysteine peptide and 53.00 % with the lysine peptide while the standard deviation (SD) of the percent peptide depletions were 1.0 % and 0.5 % for the cysteine and lysine peptides, respectively, in the valid runs. The back-calculated values of all reference control replicates were within the expected molarity concentration range for the cysteine (0.50 – 0.49 mM) and lysine peptides (0.50 – 0.49 mM) and the CV % for the nine reference controls B and C in acetonitrile were 2.6 % and 0.3 % percentages for the cysteine and lysine peptides. Thus in these runs for each peptide all validity criteria were met, confirming the validity of the assay.

Key result

Run / experiment:

other: 1

Parameter:

other: % peptide depletion cysteine

Value:

15.29

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Key result

Run / experiment:

other: 1

Parameter:

other: % peptide depletion lysine

Value:

6.62

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Key result

Run / experiment:

other: 1

Parameter:

other: % mean peptide depletion

Value:

10.96

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system:

DEMONSTRATION OF TECHNICAL PROFICIENCY:
Prior to routine use of the method, the laboratory demonstrated technical proficiency in a separate study (Study number.: 392-442-2996) by correctly obtaining the expected DPRA prediction for 10 proficiency substances as recommended in the OECD TG 442C guideline.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative (vehicle) control: yes
- Acceptance criteria met for positive control: yes
- Acceptance criteria met for variability between replicate measurements: yes

Interpretation of results:

other: positive

Conclusions:

Based on these results and the cysteine 1:10 / lysine 1:50 prediction model, the test item was concluded to be positive and have low reactivity towards the synthetic peptides thus is a potential skin sensitizer under the experimental conditions of the in chemico Direct Peptide Reactivity Assay (DPRA) method.

Executive summary:

In the course of this study the skin sensitization potential of the test item was studied using the Direct Peptide Reactivity Assay (DPRA). For the test chemical and positive control substance, in order to derive a prediction five independent tests were conducted, since the first run with cysteine peptide and the first two runs with lysine peptide did not meet the acceptance criteria and those results were rejected. All in all, the results of the two valid runs were used for the classification of the test item.

Peptide depletion resulted from the positive control cinnamaldehyde was 70.60 % with cysteine peptide and 53.00 % with the lysine peptide while the standard deviation (SD) of the percent peptide depletions were 1.0 % and 0.5 % for the cysteine and lysine peptides, respectively, in the valid runs. The back-calculated values of all reference control replicates were within the expected molarity concentration range for the cysteine (0.50 – 0.49 mM) and lysine peptides (0.50 – 0.49 mM) and the CV % for the nine reference controls B and C in acetonitrile were 2.6 % and 0.3 % percentages for the cysteine and lysine peptides. Thus in these runs for each peptide all validity criteria were met, confirming the validity of the assay. The percent cysteine peptide depletion value of the test item was 15.29 % while the percent lysine peptide depletion was 6.62 %. The mean depletion value of the peptides being 10.96 % was used to categorize the test chemical in one of the four classes of reactivity. No co-elution was observed with either cysteine or lysine peptides; therefore the cysteine 1:10 / lysine 1:50 prediction model was used for the discrimination between sensitizers and non-sensitizers. The mean peptide depletion of the test item was 10.96 %, exceeding the 6.38 % threshold of the applicable prediction model for being positive. Thus, the test item was found to be positive for skin sensitising potential under the conditions of this study.

Reference 2

Endpoint:

skin sensitisation: in vitro

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2018-04-13 to 2019-01-14

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: OECD 442E IN VITRO SKIN SENSITISATION ASSAYS ADDRESSING THE KEY EVENT ON ACTIVATION OF DENDRITIC CELLS ON THE ADVERSE OUTCOME PATHWAY FOR SKIN SENSITISATION – ANNEX I: IN VITRO SKIN SENSITISATION: HUMAN CELL LINE ACTIVATION TEST (H-CLAT) (25 JUNE 2018)

Version / remarks:

2018

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

activation of dendritic cells

Details on the study design:

Principle of the test
The h-CLAT method is an in vitro assay that quantifies changes of cell surface marker expression (i.e. CD86 and CD54) on a human monocytic leukemia cell line, THP-1 cells, following 24 hours exposure to the test item. These surface molecules are typical markers of monocytic THP-1 activation and may mimic DC activation, which plays a critical role in T-cell priming. The activation process through which DCs change from antigen processing to antigen presenting cells addresses the third key event of the skin sensitization Adverse Outcome Pathway.
The changes of surface phenotypic biomarker expression were measured and quantified by flow cytometry following cell staining with fluorochrome-tagged antibodies. The relative fluorescence intensity of surface markers compared to solvent/vehicle control were calculated and used in the prediction model, to support the discrimination between sensitisers and non-sensitisers. Cytotoxicity measurement was also conducted concurrently to assess whether upregulation of surface marker expression occurred at sub-cytotoxic concentrations.


Procedure of the h-CLAT Method
0. Doubling time and reactivity check
1. Solubility assessment of the test item (formulation)
2. Preliminary tests (dose finding tests - CV75 determination)
3. Main Tests (CD86 and CD54 expression determination)

Key result

Run / experiment:

other: result out of four runs of the individual runs for CD86

Parameter:

other:

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

other: negative

Key result

Run / experiment:

other: result out of four runs of the individual runs for CD54

Parameter:

other:

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

other: negative

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: no

DEMONSTRATION OF TECHNICAL PROFICIENCY:
Prior to routine use of the test method described in Annex I of the Test Guideline 442E, the laboratory demonstrated technical proficiency by correctly obtaining the expected h-CLAT prediction for the 10 proficiency substances recommended in OECD 442E and by obtaining CV75, EC150 and EC200 values that fell within the respective reference ranges (Study Number: 392-442-3821). Moreover, a historical database of reactivity check results positive controls and solvent/vehicle controls was generated and has been maintained to confirm the reproducibility of the test method over time.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for variability between replicate measurements: yes

In the first, third and fourth independent runs, the increase in CD86 marker expression (RFI) was not equal to or greater than 150 % at any tested dose (with >50 % of cell viability) compared to the respective negative controls. However, the increase in RFI of CD86 was greater than 150 % at most of the tested doses (with >50 % of cell viability) compared to the respective negative controls in the second run. Based on the three negative outcomes out of four individual tests, CD86 marker expression induction by the test item was concluded negative.

Regarding CD54 expression, in the first, third and fourth independent runs, the increase in CD54 marker expression (RFI) was not equal to or greater than 200 % at any tested dose (with >50 % of cell viability) compared to the respective negative controls. However, the increase in RFI of CD54 was greater than 200 % at all tested doses (with >50 % of cell viability) compared to the respective negative controls in the second run. Based on the three negative outcomes out of four individual tests, CD54 marker expression induction by the test item was concluded negative.

RFI values of concentrations with cell viabilities lower than 50 % were not taken into account when using the prediction model.

All four runs were considered valid, since all runs have met the acceptance criteria stated above.

Even though, two concordant results out of three valid runs are enough for a conclusion based on the prediction model, a fourth run was conducted to verify that the positive outcome of the second run was not specific to the test chemical but was due to an experimental artefact.

Interpretation of results:

other: negative

Conclusions:

Based on results of this OECD 442E compliant study the test item is considered negative in regards to induction of human dendritic cells.

Executive summary:

In the course of this study the skin sensitization potential of the test item was studied. The extent of cytotoxicity induced on THP-1 cells by the test item was studied in three dose finding tests. The average test item concentration that results in 75% cell viability compared to the solvent/vehicle control was 3334.1 μg/mL. This value was used for setting the dose-range for measuring CD86 and CD54 expression in the main test. Eight doses were used in four independent runs between 4010 μg/mL – 1117 μg/mL.

In the first, third and fourth independent runs, the increase in CD86 marker expression (RFI) was not equal to or greater than 150 % at any tested dose (with >50 % of cell viability) compared to the respective negative controls. However, the increase in RFI of CD86 was greater than 150 % at most of the tested doses (with >50 % of cell viability) compared to the respective negative controls in the second run. Based on the 3 negative outcomes out of 4 individual tests, CD86 marker expression induction by the test item was concluded negative.

Also, in the first, third and fourth independent runs, the increase in CD54 marker expression (RFI) was not equal to or greater than 200 % at any tested dose (with >50 % of cell viability) compared to the respective negative controls. However, the increase in RFI of CD54 was greater than 200 % at all tested doses (with >50 % of cell viability) compared to the respective negative controls in the second run. Based on the three negative outcomes out of four individual tests, CD54 marker expression induction by the test item was concluded negative.

Since the CD86 and CD54 markers gave negative results in three independent runs and gave a positive result only in one out of four runs, the overall h-CLAT prediction was concluded negative, as well.

Table 1

Name of the Test item	Obtained CV75 value (μg/mL)	h-CLAT result for CD86 (positive/negative)	h-CLAT result for CD54 (positive/negative)	h-CLAT result obtained (sensitizer/ non-sensitizer)
test item	3334.1	negative	negative	non-sensitizer

Based on these results and the h-CLAT prediction model, the test item demonstrated a non-sensitizing potential under the experimental conditions of human Cell Line Activation Test.

Reference 3

Endpoint:

skin sensitisation: in vitro

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2019-02-08 to 2019-04-25

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 442D (In Vitro Skin Sensitisation: ARE-Nrf2 Luciferase Test Method)

Version / remarks:

2018

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

activation of keratinocytes

Details on the study design:

Principle of the test
The KeratinoSens™ method is an in vitro assay that quantifies the extent of luciferase gene induction following 48 hours incubation time of the KeratinoSens™ reporter cells with the test chemicals. Luciferase gene induction is measured in the cell lysates by luminescence detection using a light producing luciferase substrate (Luciferase Reagent). Cytotoxicity and the relative luminescence intensity of luciferase substrate in the lysates are measured and luciferase induction compared to solvent/vehicle control is calculated.
KeratinoSens™ cells were derived from HaCaT human keratinocytes and transfected with selectable plasmids containing luciferase gene under the transcriptional control of the AKR1C2 ARE gene sequence, upstream of the SV40 promoter. AKR1C2 is known to be one of the genes up-regulated upon contacting skin sensitisers in dendritic cells. Therefore, this method is able to mimic the activation of the Keap1-Nrf2-ARE regulatory pathway, and is relevant for the assessment of the skin sensitisation potential of test chemicals. A prediction model is used, to support the discrimination between sensitisers and non-sensitisers.


Procedure of the KeratinoSens™ method

0. Preincubation of cells Solubility assessment of test chemicals
1. Seeding of cells for testing - 24 h incubation
2. Preparation of the stock solution
3. Preparation of master plates
4. Exposure – 48 h incubation
5. Luciferase activation measurement
6. Cytotoxicity assessment

Key result

Run / experiment:

other: overall outcome of two independent runs

Parameter:

other:

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

positive indication of skin sensitisation

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: none

DEMONSTRATION OF TECHNICAL PROFICIENCY:
Prior to routine use of the test method described in the OECD Test Guideline 442D, the laboratory demonstrated technical proficiency (Study Number: 392-442-4012), using the 10 Proficiency Substances listed in APPENDIX IA - ANNEX 1 of TG 442D. Moreover, a historical database of data generated with the positive control is maintained over time to confirm the reproducibility of the test method in the laboratory.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Acceptance criteria met for variability between replicate measurements: yes

Table 1 Summary of the KeratinoSens results for the test item

Significant induction above 1.5-fold (yes/no)	Average EC 1.5 (μM)	Average IC30 and IC50 (µM)	Average Imax (fold)	Showing clear dose response (yes/no)	KeratinoSens™ result obtained (sensitizer / non-sensitizer)
yes	325	-	18.67	yes	sensitizer

Interpretation of results:

other: positive

Conclusions:

Based on these results and the KeratinoSens™ prediction model, the test item was concluded to be positive and therefore indicating a sensitizing potential under the experimental conditions of KeratinoSens™ method (ARE-Nrf2 Luciferase Test Method).

Executive summary:

In the course of this study the skin sensitization potential of the test item was studied using the KeratinoSens™ method (ARE-Nrf2 Luciferase Test Method).

 

For the test chemical and positive control substance, in order to derive a prediction two independent tests were sufficient to be conducted, since the results of those tests were concordant and both tests met the acceptance criteria.

 

The luciferase activity induction obtained with the positive control, Trans-Cinnamaldehyde was statistically significant above the threshold of 1.5 at four concentrations in the first test and at three concentrations in the other test. The EC1.5 value of the positive control was 6 µM and 9 µM in the two individual tests, respectively. In addition, the average induction in the parallel plates for Trans Cinnamaldehyde at 64 μM was 28.35 and 83.88 fold and the dose response could be clearly seen with increasing luciferase activity induction at increasing concentrations for the positive control. There was no cytotoxicity (cell viability lower than 70 %) induced by the positive control at any of the concentrations in the first test however, there was cytotoxicity (viability < 70 %) induced at 32 and 64 µM in the second test.

 

For the test item twelve doses were used in two independent tests between 2000 µM to 1 µM. There was cytotoxicity induced (viability < 70 %) by the test item in KeratinoSens™ cells compared to the solvent/vehicle control only at the highest concentration in the second test. Therefore, no average IC30 and IC50 values were determined. The fold induction exceeded the threshold of 1.5-fold compared to the respective negative controls in both independent tests. EC1.5 values were 468 μM and 182 μM in the first and second tests respectively. Both tests were concluded positive for luciferase gene induction.

Table 1 Summary of the KeratinoSens results for the test item

Significant induction above 1.5-fold (yes/no)	Average EC 1.5 (μM)	Average IC30 and IC50 (µM)	Average Imax (fold)	Showing clear dose response (yes/no)	KeratinoSens™ result obtained (sensitizer / non-sensitizer)
yes	325	-	18.67	yes	sensitizer

Based on these results and the KeratinoSens™ prediction model, the test item was concluded to be positive and therefore indicating a sensitizing potential under the experimental conditions of KeratinoSens™ method (ARE-Nrf2 Luciferase Test Method).

 

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Additional information:

There are three in vitro/in chemico studies available with the test item assessing its skin sensitising potential. The studies were conducted according to GLP and the respective OECD TG (OECD 442C, OECD 442D, OECD 442E) and are all three considered valid. The available data is considered suitable and reliable to allow assessment of the test items properties in regards to skin sensitisation.

OECD 442C DPRA

In the course of this study the skin sensitization potential of the test item was studied using the Direct Peptide Reactivity Assay (DPRA). For the test chemical and positive control substance, in order to derive a prediction five independent tests were conducted, since the first run with cysteine peptide and the first two runs with lysine peptide did not meet the acceptance criteria and those results were rejected. All in all, the results of the two valid runs were used for the classification of the test item.

Peptide depletion resulted from the positive control cinnamaldehyde was 70.60 % with cysteine peptide and 53.00 % with the lysine peptide while the standard deviation (SD) of the percent peptide depletions were 1.0 % and 0.5 % for the cysteine and lysine peptides, respectively, in the valid runs. The back-calculated values of all reference control replicates were within the expected molarity concentration range for the cysteine (0.50 – 0.49 mM) and lysine peptides (0.50 – 0.49 mM) and the CV % for the nine reference controls B and C in acetonitrile were 2.6 % and 0.3 % percentages for the cysteine and lysine peptides. Thus in these runs for each peptide all validity criteria were met, confirming the validity of the assay. The percent cysteine peptide depletion value of the test item was 15.29 % while the percent lysine peptide depletion was 6.62 %. The mean depletion value of the peptides being 10.96 % was used to categorize the test chemical in one of the four classes of reactivity. No co-elution was observed with either cysteine or lysine peptides; therefore the cysteine 1:10 / lysine 1:50 prediction model was used for the discrimination between sensitizers and non-sensitizers. The mean peptide depletion of the test item was 10.96 %, exceeding the 6.38 % threshold of the applicable prediction model for being positive. Thus, the test item was found to be positive for skin sensitising potential under the conditions of this study.

OECD 442D KeratinoSens

In the course of this study the skin sensitization potential of the test item was studied using the KeratinoSens™ method (ARE-Nrf2 Luciferase Test Method).

 

For the test chemical and positive control substance, in order to derive a prediction two independent tests were sufficient to be conducted, since the results of those tests were concordant and both tests met the acceptance criteria.

 

The luciferase activity induction obtained with the positive control, Trans-Cinnamaldehyde was statistically significant above the threshold of 1.5 at four concentrations in the first test and at three concentrations in the other test. The EC1.5 value of the positive control was 6 µM and 9 µM in the two individual tests, respectively. In addition, the average induction in the parallel plates for Trans Cinnamaldehyde at 64 μM was 28.35 and 83.88 fold and the dose response could be clearly seen with increasing luciferase activity induction at increasing concentrations for the positive control. There was no cytotoxicity (cell viability lower than 70 %) induced by the positive control at any of the concentrations in the first test however, there was cytotoxicity (viability < 70 %) induced at 32 and 64 µM in the second test.

 

For the test item twelve doses were used in two independent tests between 2000 µM to 1 µM. There was cytotoxicity induced (viability < 70 %) by the test item in KeratinoSens™ cells compared to the solvent/vehicle control only at the highest concentration in the second test. Therefore, no average IC30 and IC50 values were determined. The fold induction exceeded the threshold of 1.5-fold compared to the respective negative controls in both independent tests. EC1.5 values were 468 μM and 182 μM in the first and second tests respectively. Both tests were concluded positive for luciferase gene induction.

Table 1 Summary of the KeratinoSens results for the test item

Significant induction above 1.5-fold (yes/no)	Average EC 1.5 (μM)	Average IC30 and IC50 (µM)	Average Imax (fold)	Showing clear dose response (yes/no)	KeratinoSens™ result obtained (sensitizer / non-sensitizer)
yes	325	-	18.67	yes	sensitizer

Based on these results and the KeratinoSens™ prediction model, the test item was concluded to be positive and therefore indicating a sensitizing potential under the experimental conditions of KeratinoSens™ method (ARE-Nrf2 Luciferase Test Method).

 

OECD 442E h-Clat

In the course of this study the skin sensitization potential of the test item was studied. The extent of cytotoxicity induced on THP-1 cells by the test item was studied in three dose finding tests. The average test item concentration that results in 75% cell viability compared to the solvent/vehicle control was 3334.1 μg/mL. This value was used for setting the dose-range for measuring CD86 and CD54 expression in the main test. Eight doses were used in four independent runs between 4010 μg/mL – 1117 μg/mL. In the first, third and fourth independent runs, the increase in CD86 marker expression (RFI) was not equal to or greater than 150 % at any tested dose (with >50 % of cell viability) compared to the respective negative controls. However, the increase in RFI of CD86 was greater than 150 % at most of the tested doses (with >50 % of cell viability) compared to the respective negative controls in the second run. Based on the 3 negative outcomes out of 4 individual tests, CD86 marker expression induction by the test item was concluded negative. Also, in the first, third and fourth independent runs, the increase in CD54 marker expression (RFI) was not equal to or greater than 200 % at any tested dose (with >50 % of cell viability) compared to the respective negative controls. However, the increase in RFI of CD54 was greater than 200 % at all tested doses (with >50 % of cell viability) compared to the respective negative controls in the second run. Based on the three negative outcomes out of four individual tests, CD54 marker expression induction by the test item was concluded negative. Since the CD86 and CD54 markers gave negative results in three independent runs and gave a positive result only in one out of four runs, the overall h-CLAT prediction was concluded negative, as well.

Table 2

Name of the Test item	Obtained CV75 value (μg/mL)	h-CLAT result for CD86 (positive/negative)	h-CLAT result for CD54 (positive/negative)	h-CLAT result obtained (sensitizer/ non-sensitizer)
test item	3334.1	negative	negative	non-sensitizer

Based on these results and the h-CLAT prediction model, the test item demonstrated a non-sensitizing potential under the experimental conditions of human Cell Line Activation Test.

Conclusion

In vitro studies covering three key steps of the adverse outcome pathway for skin sensitization as identified by the OECD (OECD Publication No.168; ENV/JM/MONO(2012)10) were performed. These study types have initially undergone validation using 54 substances (Bauch et al., 2012 Regul Toxicol Pharmacol. 63: 489-504). Recently, a validation performed with 213 substances has been published (Urbisch et al. 2015 Regul Toxicol Pharmacol. 71: 337-351).

 

Based on the results of this validation (Bauch et al., 2012 Regul Toxicol Pharmacol. 63: 489-504) the predictive capacity of the evaluation scheme using DPRA (peptide reactivity), LuSens/KeratinoSens™ (keratinocyte activation) and (m)MUSST/h-CLAT (dendritic cell activation) is comparable to that of the local lymph node assay. This was confirmed in the validation study with more substances (Urbisch 2015): When compared to the sensitization potential of the substances in humans, the classification based on an evaluation scheme using results obtained from theDPRA, LuSens and mMUSST had a sensitivity of 90%, a specificity of 90 % and an accuracy of 90 %.

 

In the evaluation scheme used here any two of the three test results determine the overall classification, i.e. any two positive test results drive the prediction of a sensitizer, while any two negative test results drive the prediction of a test substance to be a non-sensitizer (Bauch et al. 2012; Table 3). If two assays (DPRA ,LuSens or KeratinoSens, MUSST or h-CLAT) yield concordant results, the result of the third assay is not necessarily required to determine the overall outcome of the evaluation scheme.

 

Table 3: Decision matrix for combinations of DPRA, LuSens/KeratinoSens and MUSST/ h-CLAT assays.

DPRA	LuSens/ KeratinoSens^TM	MUSST/ h-CLAT	Test battery evaluation
positive	positive	positive	sensitizer
positive	positive	negative	sensitizer
positive	negative	positive	sensitizer
positive	negative	negative	non-sensitizer
negative	positive	positive	sensitizer
negative	positive	negative	non-sensitizer
negative	negative	positive	non-sensitizer
negative	negative	negative	non-sensitizer

 

In accordance with the published evaluation scheme (Bauch et al., 2012 Regul Toxicol Pharmacol. 63: 489-504) and Sections 1.2 and 1.4 of Annex XI of EC regulation 1907/2006, the test substance is considered to be a skin sensitizer based on the following results:

- DPRA: positive
- Keratinosens: positive

- h-Clat: negative

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 sensitising properties, the test item is classified and labelled as skin sensitiser Cat. 1 (H317: "May cause an allergic skin reaction") according to Regulation (EC) No 1272/2008 (CLP).