3,9-dicyclohex-3-enyl-2,4,8,10-tetraoxaspiro[5.5]undecane

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:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Principles of method if other than guideline:

The modified Local Lymph Node Assay (IMDS) was performed in 2011 on 24 female NMRI mice of the strain HsdWin:NMRI (6 animals/test item group and 6 control animals) to determine if there is any specific (sensitizing) or non-specific (irritant) stimulating potential of the test item 3,9-Di-3-cyclohexen-1-yl-2,4,8, 10-tetraoxaspiro[5.5]undecane.
A concurrent control of 6 animals treated with Alpha Hexyl Cinnamic Aldehyde was included.

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

3,9-dicyclohex-3-enyl-2,4,8,10-tetraoxaspiro[5.5]undecane

EC Number:

229-542-8

EC Name:

3,9-dicyclohex-3-enyl-2,4,8,10-tetraoxaspiro[5.5]undecane

Cas Number:

6600-31-3

Molecular formula:

C19H28O4

IUPAC Name:

3,9-bis(cyclohex-3-en-1-yl)-2,4,8,10-tetraoxaspiro[5.5]undecane

Test material form:

other: solid

Details on test material:

Test item:3,9-Di-3-cyclohexen-I-yl-2,4,8, 10-tetraoxaspiro[ 5.5] undecane
Chemical name:3,9-Di-3-cyclohexen-I-yl-2,4,8, 10-tetraoxaspiro[ 5 .5]undecane
CAS No.: 6600-31-3
Content:96.7 % (not used for calculation)
Sum formular:C19H28O4
Molar mass: 320.4 g/mol
pH-Value (diluted in water 1:9):6.2
Physical state:solid
Appearance:brown
Storage: room temperature

In vivo test system

Test animals

Species:

mouse

Strain:

NMRI

Sex:

female

Study design: in vivo (LLNA)

Vehicle:

acetone/olive oil (4:1 v/v)

Concentration:

Group 1 Vehicle (A/OO - acetone/olive oil (4:1))
Group 2 3% 3,9-Di-3-cyclohexen-I-yl-2,4,8, 10-tetraoxaspiro[ 5.5] undecane (in A/OO)
Group 3 10% 3,9-Di-3-cyclohexen-I-yl-2,4,8, 10-tetraoxaspiro[ 5.5] undecane (in A/OO)
Group 4 30% 3,9-Di-3-cyclohexen-I-yl-2,4,8, 10-tetraoxaspiro[ 5.5] undecane (in A/OO)
Group 5 30% Alpha Hexyl Cinnamic Aldehyde (in A/OO)

No. of animals per dose:

6 animals/test item group and 6 control animals

Results and discussion

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: see Remark

Remarks:

A modified Local Lymph Node Assay (IMDS) was carried out in mice. The modifications refer to the measurement of cell proliferation by cell counting instead of radioactive labeling. In addition, the acute inflammatory skin reaction is determined to discriminate specific from non-specific activation of immune competent cells in the draining lymph nodes, as also recommend in the update of OECD TG 429. 1. Direct LLNA (NMRI mice, female, 6 animalsl group) Groups Weight index Cell count index (index of mean +/-SD in %) Gr.l 1.00 +/-19.79 1.00 +/-29.46 Gr.2 0.98 +/-18.62 0.92 +/-20.44 Gr.3 0.88 +/-25.70 0.75 +/-36.18 Gr.4 0.94 +/-15.89 0.79 +/-19.54 Gr. 5 1.55 +/-21.10 1.48 +/-16.97 Ear swelling (NMRI mice, female, 6 animals/group, in 0.01 mm) Groups day 1 day4 Index day4 (mean +/- SD in %) Gr. 1 17.50 +/- 2.98 17.83+/- 3.23 1.00 Gr.2 17.33 +/- 2.84 18.17 +/- 3.18 1.02 Gr. 3 17.17 +/- 3.36 18.08 +/- 4.98 1.01 Gr. 4 17.42 +/- 2.96 18.92 +/- 5.73 1.06 Gr. 5 17.17 +/- 3.36 19.83 +/- 5.19 1.11 Ear weight (NMRI mice, female, 6 animals/group, in mg per 8 mm diameter punch) Groups day4 Index day4 (mean +/- SD in %) Gr.1 12.95 +/-5.33 1.00 Gr.2 12.96 +/- 5.47 1.00 Gr.3 12.93 +/- 8.08 1.00 Gr.4 12.73 +/- 7.85 0.98 Gr.5 14.05 +/- 5.59 1.08 Group 1 Vehicle acetone/olive oil (4:1) Group 2 3% test substance (in A/OO) Group 3 10% test substance (in A/OO) Group 4 30% test substance (in A/OO) Group 5 30% Alpha Hexyl Cinnamic aldehyde (in A/OO)

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: not applicable

Any other information on results incl. tables

Compared to vehicle-treated animals, there was no increase regarding cell counts and weights of the draining lymph nodes in the substance-treated groups. The "positive level" of index 1,4 for the cell counts has not been reached or exceeded.

The "positive level" of ear swelling, which is 2x 1 0.2 mm increase, i.e. about 10% of the control values, has not been reached or exceeded in any dose group.

Nevertheless a statistically significant increase has been detected after administration of the high concentration of the test item. Although this effect is not corroborated by increase in ear weight an irritant potential cannot fully be excluded.

These results show that there is no indication for a skin sensitizing effect after administration of a concentration up to and including 30 % 3,9-Di-3-cyclohexen-l-yl-2,4,8,10 tetraoxaspiro[5.5]undecane in this test system.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Executive summary:

The modified Local Lymph Node Assay (IMDS) was performed in 2011 on 24 female NMRI mice of the strain HsdWin:NMRI (6 animals/test item group and 6 control animals) to determine if there is any specific (sensitizing) or non-specific (irritant) stimulating potential of the test item 3,9-Di-3-cyclohexen-1-yl-2,4,8, 10-tetraoxaspiro[5.5]undecane.

A concurrent control of 6 animals treated with Alpha Hexyl Cinnamic Aldehyde was included.

The study was conducted according to OECD Guidelines No. 429 and No. 406, EC Guideline 2004173/EC (29th Adaptation of Guideline 67/S48/EEC, B,42)/Health Effects Test Guideline and OPPTS 870.2600 (EPA) with the following test item concentrations:

Test item: 0 % (vehicle control), 3 %, 10 % and 30 %.

Positive control: 30 % Alpha Hexyl Cinnamic Aldehyde

Compared to vehicle-treated animals, there was no increase regarding cell counts and weights of the draining lymph nodes in the substance-treated groups. The "positive level" of index 1,4 for the cell counts has not been reached or exceeded.

The "positive level" of ear swelling, which is 2x 1 0.2 mm increase, i.e. about 10% of the control values, has not been reached or exceeded in any dose group.

Nevertheless a statistically significant increase has been detected after administration of the high concentration of the test item. Although this effect is not corroborated by increase in ear weight an irritant potential cannot fully be excluded.

These results show that there is no indication for a skin sensitizing effect after administration of a concentration up to and including 30 % 3,9-Di-3-cyclohexen-l-yl-2,4,8,10 tetraoxaspiro[5.5]undecane in this test system.

In conclusion, these results show that the test item 3,9-Di-3-cyclohexen-l-yl-2,4,8,10 tetraoxaspiro[5.5]undecane was negative in mice after dermal application of up to and including a 30 % concentration. No indication for a non-specific (irritant) activation was detected, too. Therefore, the concentration of 30 % turned out to be the NOEL for the parameters investigated in this study with respect to skin sensitization.

These results are verified by the comparison with the results of the positive control group (Alpha Hexyl Cinnamic Aldehyde).