Registration Dossier

Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Reliable study restrictions due to read across to an analog substance

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1997
Report Date:
1997

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent

Method

Target gene:
Histidine operon (hisG46, hisC3076, hisD3052); Lipopolysaccharide barrier (LPA); DNA excision repar (uvrB)
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
mammalian microsomal enzymes
Test concentrations with justification for top dose:
with s9 mix; 25, 50, 100, 250, 1000, and 5000 ug/plate
without s9 mix: 1, 5, 10, 25, 100, 500 ug/plate
Confirmatory assay:
with s9 mix; 50, 100, 250, 500, 1000, and 5000 ug/plate
without s9 mix: 5, 10, 25, 50, 100, 500 ug/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: ethanol
- Justification for choice of solvent/vehicle:
Controls
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
other: multiple postive controls (depedning on strain and metabolic activation)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)


DURATION
- Preincubation period:
- Exposure duration: 48 hours
- Expression time (cells in growth medium):
- Selection time (if incubation with a selection agent):
- Fixation time (start of exposure up to fixation or harvest of cells):


SELECTION AGENT (mutation assays):
SPINDLE INHIBITOR (cytogenetic assays):
STAIN (for cytogenetic assays):


NUMBER OF REPLICATIONS:


NUMBER OF CELLS EVALUATED:


DETERMINATION OF CYTOTOXICITY
- Method: growth inhibition


OTHER EXAMINATIONS:
- Determination of polyploidy:
- Determination of endoreplication:
- Other:


OTHER:
Evaluation criteria:
TA98, TA100, WP2uvrA: A postive result must produce at least a 2-fold increase the mean revertants per plate of at least one tester strain over the mean revertants per plate fo the control. A dose response in the mean number of revertants per plate must also occur.
TA1535 and TA1537: A postive result must produce at least a 3-fold increase the mean revertants per plate of at least one tester strain over the mean revertants per plate fo the control. A dose response in the mean number of revertants per plate must also occur.

Results and discussion

Test results
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:
cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

EC 234-277-6, zinc bis[bis(dodecylphenyl)]bis(dithiophosphatei, s generically referred to as zinc dialkylthiophosphate (ZDDP). In vitro genotoxicity study is not available for this substance. However, sufficient data are available for a group of substances, which are produced under similar manufacturing procedures and used in commerce as multi-functional anti-wear and anti-oxidation inhibitor performance components in passenger motor oils, diesel engine oils and industrial oils such as hydraulic lubricants. They share great chemical similarity and have been considered as a category (HPV Test Plan).

CAS#

EC#

Substance Name

84605-29-8

283-392-8

Phosphorodithioic acid, mixed O,O-bis(1,3-dimethylbutyl and iso-Pr) esters, zinc salts

68457-79-4

270-608-0

Phosphorodithioic acid, mixed O,O-bis(iso-Bu and pentyl) esters, zinc salts

68909-93-3

272-723-1

Phosphorodithioic acid, mixed O,O-bis(2-ethylhexyl and iso-Pr) esters, zinc salts

4259-15-8

224-235-5

Zinc bis[O,O-bis(2-ethylhexyl)] bis(dithiophosphate)

 

In vitrobacteria gene mutation assay (AMES) has been conducted for the abovementioned substances, and the frequencies of reverse mutations in bacteria were not significantly changed after exposure to various concentrations of the registered material, with/without S9 mixture. The study is reliable without restriction (Klimitch code 1). However, for the sake of simplicity, only the AMES study on EC 224 -235 -5 is present in the current technical dossier.

CAS#

Result

84605-29-8

Negative

68909-93-3

Negative

4259-15-8

Negative

 

 Based on these results, EC 234-277-6 is expected to be non-mutagenic in bacteria.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation

No significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test material, either with or without the metabolic activation. The substance was considered to be non-mutagenic under the test conditions.