[No public or meaningful name is available]

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:

2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

publication

Title:

Safety Evaluation of a Peptide Product Derived From Sardine Protein Hydrolysates (Valtyron)

Author:

Katsuhiro Osajima et al.

Year:

2009

Bibliographic source:

International Journal of Toxicology Vol. 28 N. 5 2009 341-356

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

not specified

Remarks:

This information was not given in the publication

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Fish protein hydrolysate

IUPAC Name:

Fish protein hydrolysate

Details on test material:

Material obtained via enzymatic hydrolysis of sardine muscle

Method

Species / strain

Species / strain 1

Species / strain / cell type:

E. coli WP2 uvr A

Species / strain 2

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

Exogenous metabolic (S9) activation system (Oriental Yeast Co., Ltd., Tokyo, Japan) was prepared from the livers of male SD rats following 1 injection
of phenobarbital at 30 mg/kg BW and 3 injections of phenobarbital at 60 mg/kg BW administered every 24 hours in the morning and a single intraperitoneal injection of 5,6-benzoflavone (80 mg/kg BW) administered in the afternoon of the third day.

Test concentrations with justification for top dose:

313, 625, 1250, 2500, and 5000 ug per plate

Vehicle / solvent:

0.1Msodium-phosphate buffer (pH 7.4)

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

The 0.1-M sodiumphosphate buffer solution served as the negative control.

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

furylfuramide

other:

Details on test system and experimental conditions:

The test articles were added to the plates in a volume of 0.1 mL. In the case of assays conducted with metabolic activation, 0.5 mL of the S9 activation system was added to each plate. Three plates were used for the negative control, whereas for the positive controls and for each concentration level of the sardine peptide product tested, 2 plates were included. Plates were incubated at 37Cfor 48 hours.

Evaluation criteria:

Dose-dependent increases of greater than 2-fold compared with the negative control values were
considered a positive response.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

The sardine peptide product was determined to be nonmutagenic in the standard Ames assay in both the presence and the absence of metabolic activation. Similar results were obtained in E coli, in which the sardine peptide product also failed to induce increases in the incidence of reverse mutations.