Resin acids and Rosin acids, fumarated, esters with glycerol

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study to standard protocol subject to GLP audit.

Justification for type of information:

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

Cross-reference

Reason / purpose for cross-reference:

read-across: supporting information

Reference

Endpoint:

in vitro gene mutation study in bacteria

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 to standard protocol subject to GLP audit.

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

Key result

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

No positive results in any test strain with or without metabolic activation.

COMPARISON WITH HISTORICAL CONTROL DATA: positive and negative control data within expected values

ADDITIONAL INFORMATION ON CYTOTOXICITY:None

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Substance is clearly not genotoxic in this test

Conclusions:

Interpretation of results:
negative with metabolic activation
negative without metabolic activation

Based on an absence of genotoxic/mutagenic effects in a bacterial reverse mutation test with Salmonella typhimurium strains TA 98, TA 100, TA 1535 or TA 1537, or in E. coli strain WP2, with or without metabolic activation, Resin acids and rosin acids, fumarated, esters with pentaerythritol is not classifiable for Germ Cell Mutagenicity according to Directive 67/548/EEC, the UN Globally Harmonized System of Classification and Labelling of Chemicals (GHS) or the 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 Resin acids and Rosin acids, fumarated, esters with pentaerythritol based on category read across that is explained in the category justification document attached in Section 13 of the dossier.

In a reverse gene mutation assay in bacteria, strains TA 98, TA 100, TA 1535 and TA 1537 of S. typhimurium and strain WP2 of E. coli were exposed to resin acids and rosin acids, fumarated, esters with pentaerythritol in water/ethanol (1:19) at concentrations of 62, 185, 556, 1667 and 5000 µg/plate in the presence and absence of a mammalian metabolic activation system using both the plate incorporation method and the pre-incubation method (Vivotecnia Research, 2010). No cytotoxicity was observed at any dose level up to the limit concentration of 5000 µg/plate. No experiment with the test substance showed ratios above 2.5 as compared to the negative control, either with or without S9 metabolic activation. No dose response was observed for any of the tested bacterial strains. All vehicle and positive controls induced the appropriate responses in the corresponding strains. Based on the test conditions used in this study, resin acids and rosin acids, fumarated, esters with pentaerythritol was found to be neither mutagenic nor pro-mutagenic.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Resin acids and Rosin acids, fumarated, esters with pentaerythritol

EC Number:

305-514-1

EC Name:

Resin acids and Rosin acids, fumarated, esters with pentaerythritol

Cas Number:

94581-15-4

IUPAC Name:

Resin acids and Rosin acids, fumarated, esters with pentaerythritol

Details on test material:

- Name of test material (as cited in study report): resin acids and rosin acids, fumarated, esters with pentaerythritol
- Physical state: Solid
- Analytical purity: 100% (chemically modified UVCB)
- Lot/batch No.: PH-09/0285
- Expiration date of the lot/batch: 30/04/2010
- Storage condition of test material: -20 deg C

Method

Target gene:

His D3052
His G46
Trp
His C3076

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Additional strain / cell type characteristics:

other: All four strains uvrB, rfa and TA 98 and TA 100 were pKM 101

Metabolic activation:

with and without

Metabolic activation system:

S9 from Aroclor-induced rats.

Test concentrations with justification for top dose:

62, 185, 556, 1667 and 5000 micrograms/plate

Positive controls: 2-nitrofluorene, sodium azide, 4-nitroquinoline- N-oxide, 2-aminoanthracene, 9-aminoacridine.

Vehicle / solvent:

Vehicle: water/ethanol (1:19)
- Justification for choice of solvent/vehicle: solubility

Details on test system and experimental conditions:

Direct plate method:

Bacterial suspension, test substance and PBS (or metabolic system) (triplicates) mixed with top agar and poured onto minimal agar medium plate. After solidification, the plate was incubated at 37 deg C for 72 h.
Preincubation:

Bacterial suspension, test substance and PBS (or metabolic system) were mixed and incubated for 20 min at 37 deg C. The mixture was then plated out and incubated as described for the plate incorporation assay.


NUMBER OF REPLICATIONS: 3 for each strain and dose level in both the presence and absence of S9

Cells counted in a colony counter.

NUMBER OF CELLS EVALUATED:

Evaluation criteria:

Sterility, cytotoxicity, positive controls to show valid increases (>2.5 fold) over background, positive and negative controls within the range expected from historical data, test item to have increased >2.5 fold in revertants over control, dose response expected.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

No positive results in any test strain with or without metabolic activation.

COMPARISON WITH HISTORICAL CONTROL DATA: positive and negative control data within expected values

ADDITIONAL INFORMATION ON CYTOTOXICITY:None

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Substance is clearly not genotoxic in this test

Applicant's summary and conclusion

Conclusions:

Interpretation of results:
negative with metabolic activation
negative without metabolic activation

Based on an absence of genotoxic/mutagenic effects in a bacterial reverse mutation test with Salmonella typhimurium strains TA 98, TA 100, TA 1535 or TA 1537, or in E. coli strain WP2, with or without metabolic activation, Resin acids and rosin acids, fumarated, esters with pentaerythritol is not classifiable for Germ Cell Mutagenicity according to Directive 67/548/EEC, the UN Globally Harmonized System of Classification and Labelling of Chemicals (GHS) or the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.

Executive summary:

In a reverse gene mutation assay in bacteria, strains TA 98, TA 100, TA 1535 and TA 1537 of S. typhimurium and strain WP2 of E. coli were exposed to resin acids and rosin acids, fumarated, esters with pentaerythritol in water/ethanol (1:19) at concentrations of 62, 185, 556, 1667 and 5000 µg/plate in the presence and absence of a mammalian metabolic activation system using both the plate incorporation method and the pre-incubation method (Vivotecnia Research, 2010). No cytotoxicity was observed at any dose level up to the limit concentration of 5000 µg/plate. No experiment with the test substance showed ratios above 2.5 as compared to the negative control, either with or without S9 metabolic activation. No dose response was observed for any of the tested bacterial strains. All vehicle and positive controls induced the appropriate responses in the corresponding strains. Based on the test conditions used in this study, resin acids and rosin acids, fumarated, esters with pentaerythritol was found to be neither mutagenic nor pro-mutagenic.