Vetiveria zizanioides, ext., acetylated

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

no

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

rel-((3aR,6R)-7,7-dimethyl-8-methyleneoctahydro-1H-3a,6-methanoazulen-3-yl)methyl acetate

Molecular formula:

C17H26O2

IUPAC Name:

rel-((3aR,6R)-7,7-dimethyl-8-methyleneoctahydro-1H-3a,6-methanoazulen-3-yl)methyl acetate

Constituent 2

Reference substance name:

2-(8,8a-dimethyl-3,4,6,7,8,8a-hexahydronaphthalen-2(1H)-ylidene)propyl acetate

Molecular formula:

C17H26O2

IUPAC Name:

2-(8,8a-dimethyl-3,4,6,7,8,8a-hexahydronaphthalen-2(1H)-ylidene)propyl acetate

Constituent 3

Reference substance name:

rel-(8R,8aS)-8,8a-dimethyl-2-(propan-2-ylidene)-1,2,3,7,8,8a-hexahydronaphthalene

Molecular formula:

C15H22

IUPAC Name:

rel-(8R,8aS)-8,8a-dimethyl-2-(propan-2-ylidene)-1,2,3,7,8,8a-hexahydronaphthalene

Constituent 4

Reference substance name:

rel-(5R,10R)-6,10-dimethyl-2-(propan-2-ylidene)spiro[4.5]dec-6-en-8-one

Cas Number:

22196-46-9

Molecular formula:

C15H22O

IUPAC Name:

rel-(5R,10R)-6,10-dimethyl-2-(propan-2-ylidene)spiro[4.5]dec-6-en-8-one

Constituent 5

Reference substance name:

rel-(4R,4aS)-4,4a-dimethyl-6-(propan-2-ylidene)-4,4a,5,6,7,8-hexahydronaphthalen-2(3H)-one

Cas Number:

16981-90-1

Molecular formula:

C15H22O

IUPAC Name:

rel-(4R,4aS)-4,4a-dimethyl-6-(propan-2-ylidene)-4,4a,5,6,7,8-hexahydronaphthalen-2(3H)-one

Constituent 6

Reference substance name:

rel-(1R,8aS)-1,8a-dimethyl-7-(propan-2-ylidene)-1,2,3,7,8,8a-hexahydronaphthalene

Molecular formula:

C15H22

IUPAC Name:

rel-(1R,8aS)-1,8a-dimethyl-7-(propan-2-ylidene)-1,2,3,7,8,8a-hexahydronaphthalene

Constituent 7

Reference substance name:

2-methoxy-4-prop-1-enylphenyl acetate

EC Number:

202-236-1

EC Name:

2-methoxy-4-prop-1-enylphenyl acetate

Cas Number:

93-29-8

Molecular formula:

C12H14O3

IUPAC Name:

2-methoxy-4-(prop-1-en-1-yl)phenyl acetate

Constituent 8

Reference substance name:

rel-(1R,4aS,8aR)-1,4a-dimethyl-7-(prop-1-en-2-yl)-1,2,3,4,4a,5,6,8a-octahydronaphthalene

Molecular formula:

C15H24

IUPAC Name:

rel-(1R,4aS,8aR)-1,4a-dimethyl-7-(prop-1-en-2-yl)-1,2,3,4,4a,5,6,8a-octahydronaphthalene

Constituent 9

Reference substance name:

rel-(3aR,6R,8aR)-7,7-dimethyl-8-methylenehexahydro-1H-3a,6-methanoazulen-3(2H)-one

Cas Number:

64550-95-4

Molecular formula:

C14H20O

IUPAC Name:

rel-(3aR,6R,8aR)-7,7-dimethyl-8-methylenehexahydro-1H-3a,6-methanoazulen-3(2H)-one

Constituent 10

Reference substance name:

rel-(1R,4aS,8aR)-1-isopropyl-4,7-dimethyl-1,2,4a,5,6,8a-hexahydronaphthalene

Cas Number:

31983-21-8

Molecular formula:

C15H24

IUPAC Name:

rel-(1R,4aS,8aR)-1-isopropyl-4,7-dimethyl-1,2,4a,5,6,8a-hexahydronaphthalene

Constituent 11

Reference substance name:

6-isopropyl-4,8a-dimethyl-1,2,3,7,8,8a-hexahydronaphthalene

Cas Number:

473-14-3

Molecular formula:

C15H24

IUPAC Name:

6-isopropyl-4,8a-dimethyl-1,2,3,7,8,8a-hexahydronaphthalene

Constituent 12

Reference substance name:

rel-(4R,4aR)-4,4a-dimethyl-6-(prop-1-en-2-yl)-1,2,3,4,4a,7-hexahydronaphthalene

Molecular formula:

C15H22

IUPAC Name:

rel-(4R,4aR)-4,4a-dimethyl-6-(prop-1-en-2-yl)-1,2,3,4,4a,7-hexahydronaphthalene

Constituent 13

Reference substance name:

rel-(1R,8aR)-1-isopropyl-4,7-dimethyl-1,2,3,5,6,8a-hexahydronaphthalene

Cas Number:

317819-82-2

Molecular formula:

C15H24

IUPAC Name:

rel-(1R,8aR)-1-isopropyl-4,7-dimethyl-1,2,3,5,6,8a-hexahydronaphthalene

Constituent 14

Reference substance name:

Likely sesquiterpine, MW 202

Molecular formula:

C15H22

IUPAC Name:

Likely sesquiterpine, MW 202

Constituent 15

Reference substance name:

Likely acetylated sesquiterpenes and acetylated sesquiterpene alcohols

IUPAC Name:

Likely acetylated sesquiterpenes and acetylated sesquiterpene alcohols

Constituent 16

Reference substance name:

Likely oxygenated sesquiterpenes

IUPAC Name:

Likely oxygenated sesquiterpenes

Constituent 17

Reference substance name:

Unknown constituent, MW 260

IUPAC Name:

Unknown constituent, MW 260

Test material form:

liquid

Specific details on test material used for the study:

ROB HB = Vetiveryl Acetate
Batch: 1590849
Refrigerated (0.5 - 8 °C)
Yellow liquid

In vivo test system

Test animals

Species:

mouse

Strain:

CBA:J

Sex:

female

Details on test animals and environmental conditions:

Groups of 5 CBA/J female mice were treated.
Age Range: 11-12 weeks old.
Body weight range: 18-25 grams on Day 1.

Housed 5 mice per cage.
12 hour light/dark.
Room temperature 23-28'C.
Animals climatised for 8 days prior to the first dose.

Harlan Teklad certified rodent Chow 7012C as libitum.
Tap water ad libitum.

Study design: in vivo (LLNA)

Vehicle:

other: Diethyl phthalate/ethanol, ratio 3:1.

Concentration:

1, 2.5, 5, 10, 25%

No. of animals per dose:

5 per dose

Details on study design:

Groups of 5 female mice were treated on the dorsal surface of both ears once per day for 3 days with the test substance or the vehicle.
On day 6, the mice were injected, i.v. with 20 uCi of 3H-thymidine in sterile saline.
Five hours later the mice were euthanised and the draining auricular lymph nodes were removed.
The lymph node cells were precipitated with 5% trichloroacetic acid and the pellets counted in a scintillation counter to determine incorporation of the 3H-thymidine.

Statistics:

The EC3 was calculated using the following formula:
EC3 = c+ [(3-d) / (b-d)] (a-c)

Results and discussion

Positive control results:

NA

In vivo (LLNA)

Results

Results 1

Key result

Parameter:

SI

Value:

0.85

Variability:

Not given

Test group / Remarks:

1% - Negative

Results 2

Key result

Parameter:

SI

Value:

0.93

Variability:

Not given

Test group / Remarks:

2.5% - Negative

Results 3

Key result

Parameter:

SI

Value:

1.52

Variability:

Not given

Test group / Remarks:

10% - Negative

Results 4

Key result

Parameter:

SI

Value:

1.12

Variability:

Not given.

Test group / Remarks:

10% - Negative

Results 5

Key result

Parameter:

SI

Value:

10.18

Variability:

Not given

Test group / Remarks:

25% - Positive induction

Applicant's summary and conclusion

Interpretation of results:

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

Conclusions:

A stimulation index greater than or equal to 3 is regarded as a positive response.
Therefore, based on the criteria of the study, treatment at 25% (v/v) did result in a stimulation index of greater than 3 and hence is considered to skin sensitising activity.

Executive summary:

The EC3 was calculated to be 13.11%