Potassium hexadecyl hydrogen phosphate

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:

1987-06-24 until 1987-07-31

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1987

Report date:

1987

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1983

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Potassium hexadecyl hydrogen phosphate

EC Number:

242-768-1

EC Name:

Potassium hexadecyl hydrogen phosphate

Cas Number:

19035-79-1

Molecular formula:

C16H35O4P.K

IUPAC Name:

potassium hexadecyl hydrogen phosphate

Test material form:

liquid

Method

Target gene:

The Salmonella typhimurium histidine (his) reversion system measures his- -> his+ reversions. The Salmonella typhimurium strains are constructed to differentiate between base-pair substitution (TA 100, TA 1535, TA 102) and frameshift (TA 98, TA 1537).

Species / strain

Species / strain / cell type:

other: S. typhimurium TA 1538, TA 1535, TA 1537, TA 97, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

S9 mix (from rats induced with phenobarbital/ß-naphthoflavone)

Test concentrations with justification for top dose:

For the plate incorporation test: 10 - 1000 µg/plate
For the preincubation test: 5 - 500 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: hot water (80°C).

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

TA100, TA1535

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-acetylaminofluorene

Remarks:

TA98, TA1538

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: ICI 191

Remarks:

TA 1537 and TA 97

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

mitomycin C

Remarks:

TA 102

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-Aminoanthracene (2-AA)

Remarks:

used with all strains

Details on test system and experimental conditions:

METHOD OF APPLICATION:
- experiment I: plate incorporation test
- experiment II: pre-incubation test

DURATION
- Pre-incubation period: 30 min at 37°C (pre-incubation test)
- Expression time (cells in growth medium): 48 hours
- Number of independent experiments: 2 main experiments
- Number of replicates: 4 plates per concentration

Evaluation criteria:

The test substance was considered mutagenic if it induces at least a doubling of the number of spontaneous revertants and a dose response was recognizable. Clearly this rule of thumb has a questionable scientific foundation.

Statistics:

Mean values and standard deviations

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

Remarks:

toxic effects were visible at the highest concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

toxic effects were visible at the highest concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

toxic effects were visible at the highest concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 97

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

toxic effects were visible at the highest concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

toxic effects were visible at the highest concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 6

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

toxic effects were visible at the highest concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 7

Key result

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

toxic effects were visible at the highest concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: The test compound was dissolved in H2O at 80°C. Upon addition to the aqueous medium in the preincubation assay increasingly milky suspensions were formed starting at 50 µg/plate of the test substance.

Applicant's summary and conclusion

Conclusions:

The test substance induced no genetic damage (considered not mutagenic) in the Ames test under the described experimental conditions.

Executive summary:

Potassium hexadecylphosphate was examined for mutagenic activity in the Ames test (plate incorporation and preincubation assay). The concentration range from 10 to 1000 µg/plate in the plate incorporation assay and from 5 to 500 µg/plate in the preincubation assay was tested with seven tester strains (TA 1535, TA 1537, TA 1538, TA 97, TA 98, TA 100, TA 102). Toxic effects were visible at the highest concentrations. The tests were performed in absence as well as in presence of a phenobarbital/ß-naphthoflavone-induced rat liver homogenate fraction (S9).

The test compound was dissolved in H2O at 80°C. Upon addition to the aqueous medium in the preincubation assay increasingly milky suspensions were formed starting at 50 µg/plate of the test substance.

The test substance did not induce an increase of the frequency of revertant colonies in any of the seven tester strains used. Responsiveness of the tester strains and activity of the S9 mix were demonstrated by using appropriate positive controls for each specific strain.

Thus, it can be concluded that neither the test substance per se, nor one of its metabolites formed under the described experimental conditions induced genetic damage in the Ames test and in the preincubation modification.