Phosphoric acid, mono- and di-C16-18 (even numbered) alkyl esters, salts with 2,2'-iminodiethanol

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

S. typhimurium TA 1538

Principles of method if other than guideline:

Use of S. typhimurium strain TA 1538 instead of strain TA 102.

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Phosphoric acid, mixed decyl and octyl esters, compds. with diethanolamine

Cas Number:

68425-57-0

Molecular formula:

not applicable

IUPAC Name:

Phosphoric acid, mixed decyl and octyl esters, compds. with diethanolamine

Test material form:

solid: crystalline

Details on test material:

Name: Phosphoric acid, mixed decyl and octyl esters, compds. with diethanolamine
CAS No.: 68425-57-0
Physical state: white solid at 20 °C (turbid, non-pourable paste)

Specific details on test material used for the study:

Name: Phosphoric acid, mixed decyl and octyl esters, compds. with diethanolamine (Phosphorsaeure (C8-C10-Alkyl)-ester, DEA-Salz)
CAS No.: 68425-57-0 (68186-45-8)
Physical state: white solid at 20 °C (turbid, non-pourable paste)

Method

Target gene:

The Salmonella typhimurium histidine (his) reversion system measures his- → his+ reversions. The S. typhimurium strains are constructed to differentiate between base pair (TA 100, TA 1535) and frameshift (TA 98, TA 1537, TA 1538) mutations.

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 98

Remarks:

S. typhimurium TA 98 his D3052 rfa- uvrB- R+

Additional strain / cell type characteristics:

not specified

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 100

Remarks:

S. typhimurium TA 100 his G46 rfa- uvrB- R+

Additional strain / cell type characteristics:

not specified

Species / strain 3

Species / strain / cell type:

S. typhimurium TA 1535

Remarks:

G46 rfa- uvrB-

Additional strain / cell type characteristics:

not specified

Species / strain 4

Species / strain / cell type:

S. typhimurium TA 1537

Remarks:

his C3076 rfa- uvrB-

Additional strain / cell type characteristics:

not specified

Species / strain 5

Species / strain / cell type:

S. typhimurium TA 1538

Remarks:

his D3052 rfa- uvrB-

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9-mix

Test concentrations with justification for top dose:

The test substance was tested in two independent test series (plate incorporation assay) at concentration ranges of 8.0 - 5000 µg/plate (first test) and 6.25 - 1600 µg/plate (second test and repetition). Toxic effects were noted at concentrations of 200 µg/plate or higher.

Vehicle / solvent:

Aqua dest.

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

other: 2-aminoanthracene: TA 98 (-S9/+S9), TA 1535 (-S9/+S9), TA 100 (-S9/+S9); TA 1537 (-S9), TA 1538 (-S9/+S9); 4-Nitro-o-phenylendiamine: TA 98 (-S9/+S9), TA 1538 (-S9/+S9);

Details on test system and experimental conditions:

In order to investigate the potential of the test item for its ability to induce gene mutations the plate incorporation test (experiment I & II) was performed with the Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 1538. In two independent experiments several concentrations of the test item were used. Each assay was conducted with and without metabolic activation. The concentrations, including the controls, were tested in triplicate.
Solutions of the test substance were prepared in aqua dest. and diluted in aqua dest. just before use. The following concentrations were tested:

1st test: 8, 40, 200, 1000 and 5000 pg/plate
2nd test: 6.25, 25, 100, 400 and 1600 pg/plate (and repetition)

Evaluation criteria:

A combination of the following criteria was considered as a positive result:

- The plate background of non-reverted bacteria did not show any growth reduction versus the respective negative controls.

- The spontaneous mutation rates of each tester strain per plate were within the characteristic spontaneous mutation range.

- As a rule, the positive controls showed mutation rates exceeding the control values of TA 100 at least by the factor 2.0 and those of the other tester strains at least by the factor 3.0.

- At more than one dose tested, the test substance caused at least a 2.0-fold increase in comparison with the negative controls in the tester strain TA 100. For the other tester strains, an increase in the mutation rate of more than 3.0 above the corresponding negative controls was considered positive.

Statistics:

According to OECD guidelines, the biological relevance of the results is the criterion for the interpretation of results, a statistical evaluation of the results is not regarded as necessary.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Remarks:

Aqua dest.

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks:

Sodium azide & 2-Aminoanthracene (-S9/+S9)

Test results 2

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Remarks:

Aqua dest.

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks:

Sodium azide & 2-Aminoanthracene (-S9/+S9)

Test results 3

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Remarks:

Aqua dest.

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks:

9-Aminoacridine & 2-Aminoanthracene (-S9/+S9)

Test results 4

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks:

4-Nitro-o-phenylendiamine & 2-Aminoanthracene (-S9/+S9)

Test results 5

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Remarks:

Aqua dest.

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks:

4-Nitro-o-phenylendiamine & 2-Aminoanthracene (-S9/+S9)

Applicant's summary and conclusion

Conclusions:

In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test article did not induce point mutations by base-pair changes or frameshifts in the genome of the strains used.
Therefore, the test item is not considered to be mutagenic in this Salmonella typhimurium reverse mutation assay.

Executive summary:

In order to investigate the potential of the test item for its ability to induce gene mutations two plate incorporation tests (experiment I & II) were performed with the Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 1538. No biologically relevant increases in revertant colony numbers of any of the five tester strains were observed following treatment with the test item at any concentration level, neither in the presence nor absence of metabolic activation in experiment I and II. All criteria of validity were met.

In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test article did not induce point mutations by base-pair changes or frameshifts in the genome of the strains used. Therefore, the test item is not considered to be mutagenic in this Salmonella typhimurium reverse mutation assay.