Rosin, maleated

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: Study conducted according to guideline

Justification for type of information:

A discussion and report on the read across strategy is given as an attachment in Section 13.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

read-across: supporting information

Reference

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted according to guideline

Justification for type of information:

A discussion and report on the read across strategy is given as an attachment in Section 13.

Reason / purpose for cross-reference:

read-across source

Positive control results:

See table 1 within 'Any other information on results inc. tables' for summary of positive control data for the LLNA in the testing laboratory

Key result

Parameter:

SI

Value:

0.9

Test group / Remarks:

0.5% w/v

Key result

Parameter:

SI

Value:

3

Test group / Remarks:

5% w/v

Key result

Parameter:

SI

Value:

7.2

Test group / Remarks:

50% w/v

Key result

Parameter:

EC3

Value:

5

Test group / Remarks:

EC3 = 5%

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: 0%- 10133; 0.5% - 9402; 5%- 30450; 50%- 73071 Dpm/node (obtained dividing the dpm value by 8 - total number of lymph nodes): 1227, 1175, 3806, 9134, respectively.

There were no clinical signs of toxicity or mortality.

Bodyweight changes of the test animals between Day 0 and Day 5 were comparable to those observed in the corresponding control group animals over the same period.

Table 1. Summary of Positive Control Data for the Local Lymph Node Assay for the testing laboratory

Project number	Start date	Finish date	Test material	Concentration % (w/v)	Vehicle	Stimulation Index	Classification
039/516	22/08/01	28/08/01	4-Ethoxymethylene-2-phenyl-2-oxazolin-5-one	0.10, 0.25, 0.50	4:1 acetone/olive oil	14.01, 57.28, 42.18	Positive
039/520	12/12/01	18/12/01	Penicillin G Sodium Salt	10, 25, 50	Dimethyl Formamide	1.7, 4.1, 4.0	Positive
039/522	23/01/02	29/01/02	Polyoxyethylenesorbitan monooleate	0.25, 2.5, 25	4:1 acetone/olive oil	1.22, 2.90, 4.33	Positive
0.39/524	06/03/02	12/03/02	Eugenol	5, 25, 50	4:1 acetone/olive oil	1.7, 9.9, 0.2	Positive
039/525	07/03/02	13/03/02	Cobalt Chloride Hexahydrate	0.005, 0.05, 0.5	Dimethyl Formamide	0.7, 1.4, 6.2	Positive
039/586	13/08/02	19/08/02	Alpha-hexylcinnamaldehyde	5, 10, 50	4:1 acetone/olive oil	5.7, 5.5, 33.5	Positive

Interpretation of results:

other: Sensitising

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Under the UN Globally Harmonized System of Classification and Labelling of Chemicals (GHS) guidelines, as well as the OECD 429 test guidelines an EC3 ≥ 3 value is required to consider a substance as positive when testing using the LLNA method. Results from this study indicated that Rosin, fumarated, reaction products with formaldehyde topically administered to mice at 0.5, 5, and 50% w/v in acetone/olive oil 4:1 resulted in a pooled Stimulation Index of 0.9, 3.0, 7.2, respectively. Based on the results of this study, Rosin, fumarated, reaction products with formaldehyde is classifiable for skin sensitization according to UN Globally Harmonized System of Classification or EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.

Executive summary:

This data is being read across from the source study that tested Rosin, fumarated, reaction products with formaldehyde based on category read across that is explained in the category justification document attached in Section 13 of the dossier.

In a skin sensitization study using the Local Lymph Node Assay, groups of mice were given daily applications of 0%, 0.5%, 5%, or 50% (w/v) rosin, fumarated, reaction products with formaldehyde in acetone/olive oil 4:1 which was applied to the dorsal surface of each ear for 3 consecutive days. A control group received the vehicle only in the same manner. On Day 5, 3HTdR was injected into the tail vein, the auricular lymph nodes collected 5 hr later, and a single cell suspension prepared. 3HTdR incorporation by the cells was determined, and the Stimulation Index (SI) calculated for each dose of the test substance. The pooled Stimulation Index was 0.9, 3.0, and 7.2, respectively. Based on the results,rosin, fumarated, reaction products with formaldehyde was a skin sensitizer in mice in an OECD 429 compliant study.

Cross-reference 2

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

2003

Reference 2

Reference Type:

publication

Title:

Skin sensitization and possible groupings for "read across" for rosin based substances

Author:

Illing, HPA, Malmfors, T and Rodenburg, L

Year:

2009

Bibliographic source:

Regulatory Toxicology and Pharmacology, 54(3): 234-241

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

Rosin, fumarated, reaction products with formaldehyde

EC Number:

305-795-0

EC Name:

Rosin, fumarated, reaction products with formaldehyde

Cas Number:

95009-65-7

Molecular formula:

C25H35O6

Details on test material:

- Name of test material (as cited in study report): Rosin, formaldeyde and fumaric acid reaction product
- Substance type: Reaction product derived from UVCB
- Physical state: light brown solid
- Analytical purity: assumed 100%
- Lot/batch No.:
- Expiration date of the lot/batch:
- Storage condition of test material: deep frozen at -18C in the dark
- Other: Prepared in the laboratory as a synthesis carried out on a standard sample of rosin.

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan UK Limited, Bicester, Oxon, UK
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 15-23 g
- Housing: Individually in suspended solid-floor polypropylene cages furnished with softwood woodflakes.
- Diet (e.g. ad libitum): ad libitum (Certified rat and mouse diet - Code 5LF2- supplied by International Product Supplies Ltd, Northants, UK)
- Water (e.g. ad libitum): ad libitum (tap water)
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25
- Humidity (%): 30-70
- Air changes (per hr): approximately 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 16 October 2002 To: 22 October 2002

Study design: in vivo (LLNA)

Vehicle:

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

Concentration:

0, 0.5, 5, 50% w/v

No. of animals per dose:

4

Details on study design:

RANGE FINDING TESTS: None conducted. Information available suggested that the test material would not produce systemic toxicity or excessive local irritation at the highest suitable concentration.

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: pooled
- Criteria used to consider a positive response: SI>=3

TREATMENT PREPARATION AND ADMINISTRATION:
See further details under 'Any other information on materials and methods inc. tables'.
The method used was that described by;
Kimber et al, 1989. The murine local lymph node assay for identification of contact allergens: a preliminary evaluation of in situ measurements of lymphocyte proliferation. Contact Dermatitis 21, 215-220.
Basketter and Scholes, 1992. Comparison of the local lymph node assay with the guinea pig maximisation test for the detection of a range of contact allergens. Food and Chemical Toxicology 30, 65-69.

Positive control substance(s):

other: historical positive control data

Statistics:

no data

Results and discussion

Positive control results:

See table 1 within 'Any other information on results inc. tables' for summary of positive control data for the LLNA in the testing laboratory

In vivo (LLNA)

Results

Results 1

Key result

Parameter:

SI

Value:

0.9

Test group / Remarks:

0.5% w/v

Results 2

Key result

Parameter:

SI

Value:

3

Test group / Remarks:

5% w/v

Results 3

Key result

Parameter:

SI

Value:

7.2

Test group / Remarks:

50% w/v

Results 4

Key result

Parameter:

EC3

Value:

5

Test group / Remarks:

EC3 = 5%

Results 5

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: 0%- 10133; 0.5% - 9402; 5%- 30450; 50%- 73071 Dpm/node (obtained dividing the dpm value by 8 - total number of lymph nodes): 1227, 1175, 3806, 9134, respectively.

Any other information on results incl. tables

There were no clinical signs of toxicity or mortality.

Bodyweight changes of the test animals between Day 0 and Day 5 were comparable to those observed in the corresponding control group animals over the same period.

Table 1. Summary of Positive Control Data for the Local Lymph Node Assay for the testing laboratory

Project number	Start date	Finish date	Test material	Concentration % (w/v)	Vehicle	Stimulation Index	Classification
039/516	22/08/01	28/08/01	4-Ethoxymethylene-2-phenyl-2-oxazolin-5-one	0.10, 0.25, 0.50	4:1 acetone/olive oil	14.01, 57.28, 42.18	Positive
039/520	12/12/01	18/12/01	Penicillin G Sodium Salt	10, 25, 50	Dimethyl Formamide	1.7, 4.1, 4.0	Positive
039/522	23/01/02	29/01/02	Polyoxyethylenesorbitan monooleate	0.25, 2.5, 25	4:1 acetone/olive oil	1.22, 2.90, 4.33	Positive
0.39/524	06/03/02	12/03/02	Eugenol	5, 25, 50	4:1 acetone/olive oil	1.7, 9.9, 0.2	Positive
039/525	07/03/02	13/03/02	Cobalt Chloride Hexahydrate	0.005, 0.05, 0.5	Dimethyl Formamide	0.7, 1.4, 6.2	Positive
039/586	13/08/02	19/08/02	Alpha-hexylcinnamaldehyde	5, 10, 50	4:1 acetone/olive oil	5.7, 5.5, 33.5	Positive

Applicant's summary and conclusion

Interpretation of results:

other: Sensitising

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Under the UN Globally Harmonized System of Classification and Labelling of Chemicals (GHS) guidelines, as well as the OECD 429 test guidelines an EC3 ≥ 3 value is required to consider a substance as positive when testing using the LLNA method. Results from this study indicated that Rosin, fumarated, reaction products with formaldehyde topically administered to mice at 0.5, 5, and 50% w/v in acetone/olive oil 4:1 resulted in a pooled Stimulation Index of 0.9, 3.0, 7.2, respectively. Based on the results of this study, Rosin, fumarated, reaction products with formaldehyde is classifiable for skin sensitization according to UN Globally Harmonized System of Classification or EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.

Executive summary:

In a skin sensitization study using the Local Lymph Node Assay, groups of mice were given daily applications of 0%, 0.5%, 5%, or 50%(w/v) rosin, fumarated, reaction products with formaldehyde in acetone/olive oil 4:1 which was applied to the dorsal surface of each ear for 3 consecutive days. A control group received the vehicle only in the same manner. On Day 5, 3HTdR was injected into the tail vein, the auricular lymph nodes collected 5 hr later, and a single cell suspension prepared. 3HTdR incorporation by the cells was determined, and the Stimulation Index (SI) calculated for each dose of the test substance. The pooled Stimulation Index was 0.9, 3.0, and 7.2, respectively. Based on the results,rosin, fumarated, reaction products with formaldehyde was a skin sensitizer in mice in an OECD 429 compliant study.