Calendula officinalis, ext.

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:

25.02.2016 to 24.03.2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Calendula officinalis, ext.

EC Number:

283-949-5

EC Name:

Calendula officinalis, ext.

Cas Number:

84776-23-8

Molecular formula:

UVCB substance, not relevant.

IUPAC Name:

Calendula officinalis, ext.

Test material form:

liquid

Specific details on test material used for the study:

Teinture mère de Calendula officinalis - code C19F BATCH : M4080014

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Metabolic activation system:

S9-mix 10% (v/v)

Vehicle / solvent:

Water
Dimethyl sulfoxide (DMSO)
Acetone
NaCl 0.15 M

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

water, Dimethyl sulfoxide (DMSO), acetone, NaCl 0.15M

True negative controls:

yes

Positive controls:

yes

Positive control substance:

7,12-dimethylbenzanthracene

9-aminoacridine

2-nitrofluorene

sodium azide

other: 2-Anthramine

Details on test system and experimental conditions:

Strains of Salmonella typhimurium and Escherichia coli are obtained from MOLTOX TM . They are maintained in our laboratory.

Evaluation criteria:

Ensure that the criteria of validity of the study are well respected namely :
 the bacteriostatic activity of the highest concentration tested shall be equal to or less than 75 %,
 the spontaneous reversion rate of the absolute negative control shall comply with the historical values of the laboratory,
 the spontaneous reversion rate of the solvent shall not be statistically different from absolute negative control,
 the mean number of revertant colonies obtained for each strain and the corresponding positive control, with and/or without metabolic activation shall comply with the historical values of the laboratory.
 Negative and positive values should not show significant difference with the historical values of the laboratory (± 2 standard deviations).

Results and discussion

Test results

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks:

Positive controls induced a significant increase in the number of revertant colonies compared to negative controls.

Applicant's summary and conclusion

Conclusions:

Doses (5000, 1500, 500,150 and 50 µg/plate) prepared from solutions of the test item Teinture mère de Calendula officinalis - code C19F BATCH : M4080014 provided by Laboratoire Boiron, do not induce any mutagenic change in Salmonella typhimurium TA1535, TA1537, TA98, TA100 and in Escherichia coli WP2(uvrA¯) (pKM 101) without, or with metabolic activation, according to the OECD Guidelines n° 471. Nor hazard statement H340 is required, nor H341.