Caryophyllene, Reaction product with Acetic anhydride and acetic acid

Endpoint summary

Description of key information

VETYNAL (Sensitizing) : Key study , Local Lymph Node Assay (LLNA) in Mice (Identification of Contact Allergens)dated on 2002, performed at RCC Ltd Laboratory author Dr. W. Wang-Fan

VETYNAL EXTRA (Sensitizing): Supporting study, Local Lymph Node Assay (LLNA) in Mice (Identification of Contact Allergens) dated on 2002, performed at RCC Ltd Laboratory author Dr. W. Wang-Fan

VETYVENAL(Sensitizing): Supporting study, Local Lymph Node Assay (LLNA) in Mice (Identification of Contact Allergens)dated on 2002, performed at RCC Ltd Laboratory author Dr. W. Wang-Fan

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Reference 1

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

April 09-24 2002

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)

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Specific details on test material used for the study:

Name (as stated in the report): VETYNAL EXTRA
Batch: 9000459220
Expiration date: February 15, 2004

Species:

mouse

Strain:

CBA/Ca

Sex:

female

Details on test animals and environmental conditions:

There were 4 females per test (nulliparous and non-pregnant). At the beginning of the acclimatization, they were 7-12 weeks old at treatment and the body weights were 16.4- 19.4 g. There was one control (vehicle) group. The identification was done by unique cage number and corresponding ear number. Animals were randomly selected by computer algorithm at time of delivery. The acclimatization was done under test conditions after health examination. Only animals without any visual signs of illness were used for the study.

Vehicle:

other: ethanol:water, 7:3 (v/v)

Concentration:

The test item concentration were 0.1 %, 1 % and 10 % (w/w) in ethanol:water, 7:3 (v/v) and 100% (undiluted)

No. of animals per dose:

4

Details on study design:

The test item was placed into a glass beaker on a tared Mettler balance and the vehicle (ethanol:water, 7:3 (v/v)) was added. A weight by weight dilution was prepared using a magnetic stirrer as homogenizer. Homogeneity of the test item in the vehicle was maintained during treatment with the magnetic stirrer.

The preparations were made shortly before each dosing.

The test item was assayed at four consecutive concentrations that were selected by us.

Concentrations were in terms of material as supplied unless otherwise stated by us.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Statistics:

The mean values and standard deviations were calculated in the body weight tables.
A statistical analysis was conducted for assessment of the dose-reponse relationship, and the EC3 value was calculated according to the equation:

EC = (a-c)[(3-d)/(b-d)] + c

where EC3 is the estimated concentration of the test item required to produce a 3-fold increase in draining lymph node cell proliferative activity; (a.b) an (c,d) are respectively the co-ordinates of the two pair of data lying immediately above and below the S.I, value of 3 on the local lymph node assay dose response plot.

Positive control results:

In this study STIMULATION INDICES of 2.2, 12.0 and 6.7 were determined with the test item at concentrations of 5 %, 10 % and 25 % (w/w) in acetone:olive oil, 4:1 (v/v).

A test item is regarded as a sensitizer in the LLNA if the exposure to at least one concentration of the test item resulted in an incorporation of3HTdR at least 3-fold or greater that that recorded in control mice, as indicated by the STIMULATION INDEX (S.I.).

Based on these criteria, the test item ALPHA-HEXYLCINNAMALDEHYDE was found to be a non-sensitizer when tested up to 10% (w/w) in acetone:olive oil, 4:1 (v/v).

ALPHA-HEXYLCINNAMALDEHY showed an allergenic potency when tested at concentration of 25% in acetone:olive oil, 4:1 (v/v).

Key result

Parameter:

EC3

Value:

6

Interpretation of results:

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

Conclusions:

In this study STIMULATION INDICES of 1.0, 0.9, 4.7 and 57.3 were determined with the test item at concentrations of 0.1 %, 1 % and 10 % (w/w) in ethanol:water, 7:3 (v/v) and 100% (undiluted).

A test item is regarded as a sensitizer in the LLNA if the exposure to at least one concentration of the test item resulted in an incorporation of3HTdR at least 3-fold or greater that that recorded in control mice, as indicated by the STIMULATION INDEX (S.I.).

Based on these criteria, the test item VETYNAL EXTRA was found to be a non-sensitizer when tested up to 1% (w/w) in ethanol:water, 7:3 (v/v).

VETYNAL EXTRA showed an allergenic potency when tested at concentration of 10 % (w/w) in ethanol:water, 7:3 (v/v) and 100% (undiluted).

EC3 is the estimated concentration for a STIMULATION INDEX of 3. In this study EC3 of 6.0 % (w/w) was theoretically calculated with STIMULATION INDICES of 0.9 and 4.7 at test item concentration of 1% and 10% (w/w).

According to Basketter D. A. et al., 2002 (see Appendix B in the report, LLNA potency classification), the test item VETYNAL EXTRA could be classified as a moderate contact allergen (Class 2, EC3 value <10 %).

Executive summary:

In order to study a possible allergenic potential of VETYNAL EXTRA four groups of four female mice each were treated with the test item at concentrations of 0.1 %, 1%, 10% (w/w) in ethanol:water, 7:3 (v/v) and 100% (undiluted) by topical application to the dorsum of each ear lobe (left and right) on three consecutive days. A control group of four mice was treated with the vehicle (ethanol:water, 7:3 (v/v)) only. Five days after the first topical application the mice were injected intravenously into a tail vein with radio-labelled thymidine (³H-methylthymidine). Approximately five hours after intravenous injection, the mice were sacrificed, the draining auricular lymph node excised and pooled per group. Single cell suspension of lymph node cells were prepared from pooled lymph node which were washed subsequently and incubated with trichloroacetic acid overnight. The proliferative capacity of pooled lymph node cells was determined by the incorporation of³H-methylthymidine measured in a β-scintillation counter.

 

No test item-related clinical signs were observed.

 

All treated animals survived the scheduled study period.

 

A test item is regarded as a sensitizer in the LLNA if the exposure to at least one concentration of the test item resulted in an incorporation of³HTdR at least 3-fold or greater that that recorded in control mice, as indicated by the STIMULATION INDEX (S.I.).

Reference 2

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April 9-24 2002

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)

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Specific details on test material used for the study:

Name (as stated in the report): VETYNAL
Batch: VE00473596
Expiration date: February 05, 2003

Species:

mouse

Strain:

CBA/Ca

Sex:

female

Details on test animals and environmental conditions:

There were 4 females per test (nulliparous and non-pregnant). At the beginning of the acclimatization, they were 7-13 weeks old at treatment and the body weight were 17.5–20.4 g. There was one control (vehicle) group. The identification was done by unique cage number and corresponding ear number. Animals were randomly selected by computer algorithm at time of delivery. The acclimatization was done under test conditions after health examination. Only animals without any visual signs of illness were used for the study.

Vehicle:

other: ethanol:water, 7:3 (v/v)

Concentration:

The test item concentration were 0.1 %, 1 % and 10 % (w/w) in ethanol:water, 7:3 (v/v) and 100% (undiluted)

No. of animals per dose:

4

Details on study design:

The test item was placed into a glass beaker on a tared Mettler balance and the vehicle (ethanol:water, 7:3 (v/v)) was added. A weight by weight dilution was prepared using a magnetic stirrer as homogenizer. Homogeneity of the test item in the vehicle was maintained during treatment with the magnetic stirrer.

The preparations were made shortly before each dosing.

The test item was assayed at four consecutive concentrations that were selected by us.

Concentrations were in terms of material as supplied unless otherwise stated by us.

Positive control substance(s):

not specified

Statistics:

The mean values and standard deviations were calculated in the body weight tables.
A statistical analysis was conducted for assessment of the dose-reponse relationship, and the EC3 value was calculated according to the equation:

EC = (a-c)[(3-d)/(b-d)] + c

where EC3 is the estimated concentration of the test item required to produce a 3-fold increase in draining lymph node cell proliferative activity; (a.b) an (c,d) are respectively the co-ordinates of the two pair of data lying immediately above and below the S.I, value of 3 on the local lymph node assay dose response plot.

Key result

Parameter:

EC3

Value:

12.4

Interpretation of results:

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

Conclusions:

In this study STIMULATION INDICES of 1.3, 1,1, 1.8 and 47.1 were determined with the test item at concentrations of 0.1 %, 1 % and 10 % (w/w) in ethanol:water, 7:3 (v/v) and 100% (undiluted).

A test item is regarded as a sensitizer in the LLNA if the exposure to at least one concentration of the test item resulted in an incorporation of3HTdR at least 3-fold or greater that that recorded in control mice, as indicated by the STIMULATION INDEX (S.I.).

Based on these criteria, the test item VETYNAL was found to be a non-sensitizer when tested up to 10% (w/w) in ethanol:water, 7:3 (v/v).

VETYNAL showed an allergenic potency when tested at concentration of 100% (undiluted). EC3 is the estimated concentration for a STIMULATION INDEX of 3. In this study EC3 of 12.4 % (w/w) was theoretically calculated with STIMULATION INDICES of 1.8 and 47.1 at test item concentration of 10% (w/w) and 100% (undiluted).

According to Basketter D. A. et al., 2002 (see Appendix B in the report, LLNA potency classification), the test item VETYNAL could be classified as a weak contact allergen (Class 3, EC3 value 10–30 %).

Executive summary:

In order to study a possible allergenic potential of VETYNAL four groups of four female mice each were treated with the test item at concentrations of 0.1 %, 1%, 10% (w/w) in ethanol:water, 7:3 (v/v) and 100% (undiluted) by topical application to the dorsum of each ear lobe (left and right) on three consecutive days. A control group of four mice was treated with the vehicle (ethanol:water, 7:3 (v/v)) only. Five days after the first topical application the mice were injected intravenously into a tail vein with radio-labelled thymidine (³H-methylthymidine). Approximately five hours after intravenous injection, the mice were sacrificed, the draining auricular lymph node excised and pooled per group. Single cell suspension of lymph node cells were prepared from pooled lymph node which were washed subsequently and incubated with trichloroacetic acid overnight. The proliferative capacity of pooled lymph node cells was determined by the incorporation of ³H-methylthymidine measured in a β-scintillation counter.

No test item-related clinical signs were observed.

 

All treated animals survived the scheduled study period.

 

A test item is regarded as a sensitizer in the LLNA if the exposure to at least one concentration of the test item resulted in an incorporation of ³HTdR at least 3-fold or greater that that recorded in control mice, as indicated by the STIMULATION INDEX (S.I.).

Reference 3

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

April 09-24 2002

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)

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Specific details on test material used for the study:

Name (as stated in the report): VETYVENAL
Batch: 9000387635
Expiration date: June 19, 2002

Species:

mouse

Strain:

CBA/Ca

Sex:

female

Details on test animals and environmental conditions:

There were 4 females per test (nulliparous and non-pregnant). At the beginning of the acclimatization, they were 7-12 weeks old at treatment and the body weight were 17.0- 20.6 g. There was one control (vehicle) group. The identification was done by unique cage card and individual color code. Animals were randomly selected by computer algorithm at time of delivery. The acclimatization was done under test conditions after health examination. Only animals without any visual signs of illness were used for the study.

Vehicle:

other: ethanol:water, 7:3 (v/v)

Concentration:

The test item concentration were 0.1%, 1%, 10% (w/w) in ethanol:water, 7:3 (v/v) and 100% (undiluted)

No. of animals per dose:

4

Details on study design:

The test item was placed into a glass beaker on a tared Mettler balance and the vehicle (ethanol:water, 7:3 (v/v)) was added. A weight by weight dilution was prepared using a magnetic stirrer as homogenizer. Homogeneity of the test item in the vehicle was maintained during treatment with the magnetic stirrer.

The preparations were made shortly before each dosing.

The test item was assayed at four consecutive concentrations that were selected by us.

Concentrations were in terms of material as supplied unless otherwise stated by us.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Statistics:

The mean values and standard deviations were calculated in the body weight tables.
A statistical analysis was conducted for assessment of the dose-reponse relationship, and the EC3 value was calculated according to the equation:

EC = (a-c)[(3-d)/(b-d)] + c

where EC3 is the estimated concentration of the test item required to produce a 3-fold increase in draining lymph node cell proliferative activity; (a.b) an (c,d) are respectively the co-ordinates of the two pair of data lying immediately above and below the S.I, value of 3 on the local lymph node assay dose response plot.

Positive control results:

In this study STIMULATION INDICES of 2.2, 2.0 and 6.7 were determined with the test item at concentrations of 5 %, 10 % and 25 % (w/w) in acetone:olive oil, 4:1 (v/v).
A test item is regarded as a sensitizer in the LLNA if the exposure to at least one concentration of the test item resulted in an incorporation of3HTdR at least 3-fold or greater that that recorded in control mice, as indicated by the STIMULATION INDEX (S.I.).

Based on these criteria, the test item ALPHA-HEXYLCINNAMALDEHYDE was found to be a non-sensitizer when tested up to 10% (w/w) in acetone:olive oil, 4:1 (v/v).

ALPHA-HEXYLCINNAMALDEHY showed an allergenic potency when tested at concentration of 25% in acetone:olive oil, 4:1 (v/v).

Key result

Parameter:

EC3

Value:

12.9

Interpretation of results:

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

Conclusions:

In this study STIMULATION INDICES of 0.9, 0.7, 1.8 and 38.9 were determined with the test item at concentrations of 0.1 %, 1 % and 10 % (w/w) in ethanol:water, 7:3 (v/v) and 100% (undiluted).

A test item is regarded as a sensitizer in the LLNA if the exposure to at least one concentration of the test item resulted in an incorporation of3HTdR at least 3-fold or greater that that recorded in control mice, as indicated by the STIMULATION INDEX (S.I.).

Based on these criteria, the test item VETYNAL EXTRA was found to be a non-sensitizer when tested up to 10% (w/w) in ethanol:water, 7:3 (v/v).

VETYVENAL showed an allergenic potency when tested at concentration of 100% (undiluted).

EC3 is the estimated concentration for a STIMULATION INDEX of 3. In this study EC3 of 12.9 % (w/w) was theoretically calculated with STIMULATION INDICES of 0.9 and 4.7 at test item concentration of 10% (w/w) and 100% (undiluted).

According to Basketter D. A. et al., 2002 (see Appendix B in the report, LLNA potency classification), the test item VETYNAL EXTRA could be classified as a moderate contact allergen (Class 3, EC3 value 10-30 %).

Executive summary:

In order to study a possible allergenic potential of VETYVENAL four groups of four female mice each were treated with the test item at concentrations of 0.1 %, 1%, 10% (w/w) in ethanol:water, 7:3 (v/v) and 100% (undiluted) by topical application to the dorsum of each ear lobe (left and right) on three consecutive days. A control group of four mice was treated with the vehicle (ethanol:water, 7:3 (v/v)) only. Five days after the first topical application the mice were injected intravenously into a tail vein with radio-labelled thymidine (3H-methylthymidine). Approximately five hours after intravenous injection, the mice were sacrificed, the draining auricular lymph node excised and pooled per group. Single cell suspension of lymph node cells were prepared from pooled lymph node which were washed subsequently and incubated with trichloroacetic acid overnight. The proliferative capacity of pooled lymph node cells was determined by the incorporation of3H-methylthymidine measured in a β-scintillation counter.

 

No test item-related clinical signs were observed.

 

All treated animals survived the scheduled study period.

 

A test item is regarded as a sensitizer in the LLNA if the exposure to at least one concentration of the test item resulted in an incorporation of3HTdR at least 3-fold or greater that that recorded in control mice, as indicated by the STIMULATION INDEX (S.I.).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available