Insulin Human Methyl Ester

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:

December 1985

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1985

Report date:

1985

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Human Insulin

Molecular formula:

C257H383N65O77S6

IUPAC Name:

Human Insulin

Test material form:

solid: particulate/powder

Remarks:

White powder

Details on test material:

Molecular formula: C257H383N65O77S6
Molecular weight: 5807.66 g/mol

Specific details on test material used for the study:

NA

Method

Species / strain

Species / strain / cell type:

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

Additional strain / cell type characteristics:

other: Histidine dependent

Metabolic activation:

with and without

Metabolic activation system:

Mammalian liver post-mitochondrial fraction (S-9) prepared from rats induced with Arochlor 1254

Test concentrations with justification for top dose:

Two experiments were carried out in triplicates:

Range finder experiment and mutation experiment 1:
Test concentrations: 40 - 4000 ug/mL

Mutation experment 2:
Test concentrations: 40 - 4000 ug/mL

Vehicle / solvent:

Due to the proteineous nature of the test substance, sterile water was considered to be the most appropriate solvent for this study.

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

benzo(a)pyrene

other: 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

Range-finder experiment:
S. typhimurium TA100 were exposed in triplicates to the above mentioned concentrations with and without S-9 (4 solvent controls and triplicate positive controls) for 3 days. Plates were investigated for toxicity and where possible revertant colonies were counted.

Mutation experiments:
All four strains were tested in triplicates with and without S-9.

Preincubation: overnight

Evaluation criteria:

Not specified

Statistics:

Not specified

Results and discussion

Test results

Key result

Species / strain:

S. typhimurium, other: TA98, TA100, TA1535, TA1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

Mutagenicity of Human Insulin (API) was determined according to OECD TG 471. The test substance did not induce mutation in Salmonella typhimurium both in the absence and in the presence of a rat liver metabolic activation system (S-9).

Executive summary:

Mutagenicity of Human Insulin (API) was determined according to OECD TG 471. The test substance did not induce mutation in four histidine-requiring strains of Salmonella typhimurium (TA98, TA100, TA1535, and TA1537), when tested under conditions employed in this study. The conditions included treatments at concentrations up to 4000 ug/plate, both in the absence and in the presence of a rat liver metabolic activation system (S-9) in two separate experiments.