Oct-1-ene-3-ol

Endpoint summary

Description of key information

In 2015 both an In vitro Direct peptide reactivity assay (DPRA) and an In vitro KeratinoSens assay were run on the test material. Both In vitro assays resulted in positive results. Consequently, the substance was classified as a Sensitiser.

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:

Study carried out from 9 November 2015 to 26 November 2015

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))

Deviations:

no

Principles of method if other than guideline:

The Direct Peptide Reactivity Assay (DPRA) is an in chemico test to determine the reactivity of test a substance towards peptides.
This assay has been validated for a broad range of low-molecular weight chemicals and it was found to detect reactive skin sensitizers from a broad range of so called applicability domains, i.e. chemicals reacting with proteins by different mechanisms. It was validated by ECVAM and proposed to be used as part of an integrated approach for testing and assessment (IATA).

GLP compliance:

yes

Type of study:

direct peptide reactivity assay (DPRA)

Specific details on test material used for the study:

Lot No.: VE00345297
Physical form: Liquid
Purity: 99.8%

Details on the study design:

The test substance AMYL VINYL CARBINOL was dissolved in acetonitrile and mixed with a Cysteine- and a Lysine-containing peptide according to the standard operating procedure of the DPRA. One study with three replicates was conducted. After 24 h incubation time, peptide depletion induced by AMYL VINYL CARBINOL was determined by HPLC-UV.

Positive control results:

As indicated in Table 7, all the acceptance criteria were fulfilled for the positive control cinnamic aldehyde, with the exception of the CV for the Lys peptide which is at 12.4%, and thus slightly above the threshold of 11.6. However since this is so close to the threshold and the three individual depletion values (64.6, 50.9, 62.6) all nicely fall in the target range and the historical range (Table C1), this was considered an acceptable deviation.

Parameter:

other: Cys-peptide depletion

Value:

65.2

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of skin sensitisation

Remarks:

Value is a percentage

Parameter:

other: Lys-peptide depletion

Value:

1.2

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of skin sensitisation

Remarks:

Value is a percentage

Other effects / acceptance of results:

The test substance gave 65.2 % depletion of the Cys-peptide and 6.7 % depletion of the Lys-peptide. The average peptide depletion is 36.0 %. This is below the threshold of 22.6%, and the substance is thus attributed to the “moderate” reactivity class, rating it as a sensitizer according to the DPRA prediction model.
Acceptance criteria: The standard deviation for Cys-peptide depletion should be < 14.9% and for Lys-peptide depletion < 11.6%. These criteria were fulfilled (4.7% and 0.3% SD, respectively).
The co-elution controls indicated no co-elution with an UV-absorbing component.

	Average	Standard deviation
Cys-peptide depletion	65.2	4.7
Lys-peptide depletion	6.7	0.3
Average depletion Cys-and Lys-peptide	36.0
Reactivity Class	MODERATE
Prediction	Sensitizer
Elution time Cys peptide	10.75
Elution time test substance Cys peptide run	No UV peak
Elution time Lys peptide	8.02
Elution time test substance Lys peptide run	No UV peak

Interpretation of results:

Category 1B (indication of skin sensitising potential) based on GHS criteria

Conclusions:

The result of the DPRA assay should be used as part of an integrated approach for testing and assessment (IATA). A parallel test in the KeratinoSens™ assay may indicate whether congruent results are obtained by both test methods. According to a detailed analysis on large set of chemicals, two congruent results in these two tests give a good prediction of the sensitizer hazard [3-5] particularly when predicting human data, while an additional test in a dendritic cell line assessing expression of surface markers may be needed in case of discordant results.
AMYL VINYL CARBINOL was peptide-reactive and classified into the moderate reactivity class according to the prediction model. It is therefore considered a sensitizer according to the prediction model of the DPRA.

Executive summary:

AMYL VINYL CARBINOL was peptide-reactive and classified into the moderate reactivity class according to the prediction model. It is therefore considered a sensitizer according to the prediction model of the DPRA.

Reference 2

Endpoint:

skin sensitisation: in vitro

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Study carried out from 16 November 2015 to 26 November 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study carried out according to guidelines.

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Principles of method if other than guideline:

The KeratinoSensTM assay is a cell-based assay with a reporter cell line to detect potential skin sensitizers by their ability to induce the Nrf2-response.
This assay has been validated for a broad range of low-molecular weight chemicals and it was found to respond to skin sensitizers from a broad range of so called applicability domains, ie. chemicals reacting with proteins by different mechanisms. It was validated by ECVAM and proposed to be used as part of an integrated approach for testing and assessment (IATA).

GLP compliance:

yes

Type of study:

activation of keratinocytes

Specific details on test material used for the study:

Lot No.: VE00345297
Physical form: Liquid
Purity: 99.8%

Details on the study design:

The Cosmetic directive is phasing out of animal testing for new products. This has led to the development of potential alternative assays to screen for sensitizing potential. These new assays were tested against a large list of reference chemicals in order to prove their applicability domain.
The KeratinoSensTM assay is a cell-based assay with a reporter cell line to detect potential skin sensitizers by their ability to induce the Nrf2-response [2].
This assay has been validated for a broad range of low-molecular weight chemicals [2-8] and it was found to respond to skin sensitizers from a broad range of so called applicability domains, i.e. chemicals reacting with proteins by different mechanisms.
The assay underwent validation at the European Centre for the Validation of Alternative Methods (ECVAM) [9] and an OECD test guideline was adopted (OECD TG 442d). The protocol was published as DB-ALM protocol 155 [1]. The assay was proposed by ECVAM [9] to be to be used as part of an integrated approach for testing and assessment (IATA).

Positive control results:

Cinnamic aldehyde was run in all three repetitions. Here the detailed results for this positive control are reported in Table 8 and Figure 5. Cinnamic aldehyde needs to be positive for a run to be accepted (i.e. induction > 1.5 fold). This was the case in all three repetitions. The induction at 64 µM and the EC 1.5 for cinnamic aldehyde were also calculated. The targets are: (i) Average induction in the three replicates for cinnamic aldehyde at 64 µM should be between 2 and 8, and (ii) the EC 1.5 value should be between 7 µM and 30 µM. At least one of these two numerical criteria must be met in order to accept a repetition. In the experiments performed here both criteria were fulfilled in all three repetitions. Thus all three repetitions were valid for the positive control.

Run / experiment:

other: Rep 1

Parameter:

other: Cytotoxicity

Value:

1 000

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of skin sensitisation

Remarks:

Result Value in μM

Run / experiment:

other: Rep 2

Parameter:

other: Cytotoxicity

Value:

1 000

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of skin sensitisation

Remarks:

Result Value in μM

Run / experiment:

other: Rep 3

Parameter:

other: Cytotoxicity

Value:

835.23

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of skin sensitisation

Remarks:

Result Value in μM

Run / experiment:

other: Rep 1

Parameter:

other: Luciferase

Value:

3.11

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of skin sensitisation

Run / experiment:

other: Rep 2

Parameter:

other: Luciferase

Value:

2.56

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of skin sensitisation

Remarks:

Result Value is 'Imax fold-induction'

Run / experiment:

other: Rep 3

Parameter:

other:

Value:

5.78

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of skin sensitisation

Remarks:

Result Value is 'Imax fold-induction'

Cytotoxicity determinations. Given is the IC50 value as the concentration in µM reducing the viability by 50%.

	Test substance	Rep 1 IC50 (µM)(*)	Rep 2 IC50 (µM)	Rep 3 IC50 (µM)	Geometric Mean IC 50 (µM)	Standard deviation IC 50 (µM)
1	AMYL VINYL CARBINOL	>1000	>1000	835.23	>1000

Luciferase determinations. Given is the I_maxvalues indicating maximal fold-induction up to a concentration of 1000µM.

	Test substance	Rep 1 IMAX(fold induction)	Rep 2 IMAX(fold induction)	Rep 3 IMAX(fold induction)	Average IMAX(fold induction)	Standard deviation IMAX(fold induction)
1	AMYL VINYL CARBINOL	3.11	2.56	5.78	3.81	1.72

Luciferase determinations.Given is the EC1.5, EC2 and EC3 value as the concentration in s indicating maximal fold-induction up to a concentration of 1000µM.

	Test substance	Extrapolated value	Rep 1   (µM)	Rep 2    (µM)	Rep 3   (µM)	Geometric Mean   (µM)	Standard deviation   (µM)
1	AMYL VINYL CARBINOL	EC 1.5	398.26*	545.7	367.61	430.69	95.21
1	AMYL VINYL CARBINOL	EC 2	590.36	760.51	531.27	620.18	119.02
1	AMYL VINYL CARBINOL	EC 3	959.19	n.i.	655.43	792.90	214.79

* induction at slight cytotoxic value (67% viability), Repetition 1 is negative according the prediction model; n.i. no induction above a given threshold.

Overall rating of the test substance according to the prediction model and the number of positive repetitions.

		Reps pos.	Overall rating
1	AMYL VINYL CARBINOL	2 of 3	POSITIVE

Interpretation of results:

Category 1B (indication of skin sensitising potential) based on GHS criteria

Conclusions:

In all three repetitions, induction of the luciferase above the threshold of 1.5-fold was noted, and in two repetitions this induction was in the non-cytotoxic range. According to the prediction model of the KeratinoSens™ assay, the test substance is rated as a sensitizer. However, this response only occurs at relatively high levels, indicating a weak potency. This is also clearly supported by the analysis of the dose-response curve in Figure 4 with overall clear induction of the luciferase reporter gene to be observed at high test concentrations.

Executive summary:

The KeratinoSensTM assay is a cell-based assay with a reporter cell line to detect potential skin sensitizers by their ability to induce the Nrf2-response.

This assay has been validated for a broad range of low-molecular weight chemicals and it was found to respond to skin sensitizers from a broad range of so called applicability domains, i.e. chemicals reacting with proteins by different mechanisms. It was validated by ECVAM and proposed to be used as part of an integrated approach for testing and assessment (IATA).

The test substance Amyl Vinyl Carbinol was dissolved in DMSO and tested according to the standard operating procedure of the KeratinoSensTM assay at 12 concentrations in three repetitions, each time in three replicates. After 48 h incubation time, luciferase induction and cellular viability at each of the concentrations were determined.

Amyl Vinyl Carbinol was non-toxic to the KeratinoSens™ cells. In all three repetitions, it did induce the luciferase gene above a threshold of 1.5-fold with an average EC1.5 value of 430 μM. It is therefore considered a sensitizer according to the prediction model of the KeratinoSens™ assay.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Justification for classification or non-classification

In 2015 both an In vitro Direct peptide reactivity assay (DPRA) according to OECD Guideline 442C (In Chemico Skin Sensitisation: Direct Peptide Reactivity Assay (DPRA)) and an In vitro KeratinoSens assay according to OECD Guideline 442D (In Vitro Skin Sensitisation: ARE-Nrf2 Luciferase Test Method) were run on the test material. Both In vitro assays resulted in positive results. Consequently, the substance was classified as a sensitiser.