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Toxicological information

Genetic toxicity: in vitro

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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

Constituent 1
Chemical structure
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.