Phosphatidylcholines, soya, hydrogenated

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:

1995.07.28

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Phosphatidylcholines, soya, hydrogenated

EC Number:

306-549-5

EC Name:

Phosphatidylcholines, soya, hydrogenated

Cas Number:

97281-48-6

Molecular formula:

C44H88NO8P

IUPAC Name:

[(2R)-2,3-di(octadecanoyloxy)propyl] 2-(trimethylazaniumyl)ethyl phosphate

Test material form:

solid

Specific details on test material used for the study:

Batch no. 42000100
white crystalline powder
expiry date 7/1996

Method

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Species / strain 3

Species / strain / cell type:

E. coli WP2 uvr A pKM 101

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

10-10.000 µg/plate. No bacteriotoxic effects were observed however precipitation in the range from 1000-10.000 µg/plate therefore the following concentrations were applied:
8, 40, 200, 1000 and 5000 µg/plate

Vehicle / solvent:

ethanol

Controls

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

N-ethyl-N-nitro-N-nitrosoguanidine

other: 4-nitro-1,2-phenylene diamine and 2-aminoanthracene

Details on test system and experimental conditions:

The positive controls (without the S 9 mix):
sodium azide (10µg/plate used for TA 1535 and TA 100) ,
9-aminoacridine hydrochloride (50µg/plate used for TA 1537)
4-nitro-1,2 phenylene diamine (10µg/plate used for TA 1537 and TA 98)
and N-ethyl N-nitro nitrosguanidin (5 µg/plate used for E. coli WP2 uvrA and pKM101)

The positive control 2-aminoantracene (3 and 40 µg/plate) was used for positive control for tests with S9 metabolic activation
Negative control: Solvent without test article

Evaluation criteria:

Criteria for acceptance of assay:
- The negative controls had to be within the expected range as defined by published data (Maron and Ames 1983)
- The positive controls had to show sufficient effects as defined by the laboratory's experience
- The titer determination had to reveal a sufficient bacterial density in the suspension

Assessment of mutagenicity and baceriotoxicity:
A reproducible and dose-related increase of mutants counts for at least one strain is considered positive
For TA 98, TA 1535, WP2 uvrA and WP2 CM a twofold increase of revernants compared to the negative controls should be reached, whereas for TA 1537 a threefold increase should be attained. For TA 100 a 1.5-fold increase is regarded as an indication of potential mutagenicity. Otherwise the results are considered to be negative.

The criterion for a biologically significant bacteriotoxic effect is a reduction in the number of colonies/plate or revernants/plate or in background growth by more than 50% relative to the respective negative control.

Statistics:

NA

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Positive controls validity:

valid

Test results 6

Key result

Species / strain:

E. coli WP2 uvr A pKM 101

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Positive controls validity:

valid

Additional information on results:

Hydrogenated soya phosphatidylcholine caused no mutagenic effects at concentrations ranging from 8-5000 µg/plate

Remarks on result:

other: No mutagenic nor bacteriotoxic effects at concentrations ranging from 8-5000 µg/plate

Applicant's summary and conclusion

Conclusions:

Hydrogenated soya phosphatidylcholine caused no mutagenic effects at concentrations ranging from 8-5000 µg/plate.

Executive summary:

Hydrogenated soya phosphatidylcholine was investigated in the Salmonella typhimurium (Ames test) and Escherichia coli Reverse mutation Assay for point mutations using Salmonella Typhimurium LT2 mutants and two E.coli WP2 mutants. These two tester strains were the histidine auxotrophic Salmonella strains TA1535, TA 1537, TA 98, Ta 100 and the trypto auxotrophic E. coli strains WP2 uvrA and WP2 uvr (pkM101). The study was conducted according to the OECD Guidelines 471/472 and following GLP.

Hydrogenated soya phosphatidylcholine caused no mutagenic effects at concentrations ranging from 8-5000 µg/plate.

In the positive control the mutant counts increased to more than twice the value of the negative controls, demonstrating that the system was highly sensitive.