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Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information
Sodium fluorescein is non mutagenic in the Ames test performed for the Salmonella typhimurium TA1538, TA98, and TA100,TA 1537 and TA 1535.
Link to relevant study records
Reference
Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Reference:
Composition 0
Principles of method if other than guideline:
Mutagenecity study was conducted to study the effects of sodium fluorescein on bacterial strains using Ames test.
GLP compliance:
not specified
Type of assay:
bacterial gene mutation assay
Test material information:
Composition 1
Species / strain:
other: TA 1538, TA98, TA1537, TA1535, and TA100
Details on mammalian cell lines (if applicable):
Not applicable
Additional strain characteristics:
not applicable
Test concentrations with justification for top dose:
10mg/plate
Vehicle:
- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: No data available
Negative controls:
not specified
Solvent controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
2-acetylaminofluorene
4-nitroquinoline-N-oxide
benzo(a)pyrene
Details on test system and conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: No data available
- Exposure duration: No data available
- Expression time (cells in growth medium): No data available
- Selection time (if incubation with a selection agent): No data available
- Fixation time (start of exposure up to fixation or harvest of cells): No data available

SELECTION AGENT (mutation assays): No data available
SPINDLE INHIBITOR (cytogenetic assays): No data available
STAIN (for cytogenetic assays): No data available

NUMBER OF REPLICATIONS: No data available

NUMBER OF CELLS EVALUATED: No data available

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: Partial lethality observed

OTHER EXAMINATIONS:
- Determination of polyploidy: No data available
- Determination of endoreplication: No data available
- Other: No data available

OTHER: No data available
Evaluation criteria:
Based on Dose related increase in His revertants
Statistics:
No data
Species / strain:
other: TA1538, TA98, and TAI00,TA 1537 and TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Remarks:
Partialy lethal
Vehicle controls valid:
yes
Negative controls valid:
not specified
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No data available
- Effects of osmolality: No data available
- Evaporation from medium: No data available
- Water solubility: No data available
- Precipitation: No data available
- Other confounding effects: No data available

RANGE-FINDING/SCREENING STUDIES: No data available

COMPARISON WITH HISTORICAL CONTROL DATA: No data available

ADDITIONAL INFORMATION ON CYTOTOXICITY: Partial lethality was observed by reductions in the His- background in some bacterial strains at the dose of 10mg/plate.
Conclusions:
Interpretation of results (migrated information):
negative

Sodium fluorescein is non mutagenic in the Ames test performed for the Salmonella typhimurium TA1538, TA98, and TA100,TA 1537 and TA 1535.
Executive summary:

The fluorescent dyeSodium fluorescein issold for public, commercial, and field use, in liquid and tablet forms, for sewage, plumbing, and water pollution tracing. Ames test was conducted to evaluate the mutagenic capability of sodium fluorescein. The dose concentration used was 10mg/plate using water as vehicle. No increase in number of revertants observed indicating the substance to be non mutagenic with or without activation. Sodium fluorescein is non mutagenic in the Ames test performed for theSalmonella typhimuriumTA1538, TA98, and TA100,TA 1537 and TA 1535.

 

In accordance with CLP classification, the test material does not classify as a mutagen.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

Various studies for the target chemical were reviewed to summarize the following information:

 

An in vitro mammalian cell gene mutation study was designed by Eagle R. Nestmann , David J. Kowbel And Jennmm Ellenton (1980) and conducted to determine the genotoxicity profile of sodium fluorescein (CAS No. 518-47-8). The fluorescent dye Sodium fluorescein is sold for public, commercial, and field use, in liquid and tablet forms, for sewage, plumbing, and water pollution tracing. Ames test was conducted to evaluate the mutagenic capability of sodium fluorescein. The dose concentration used was 10mg/plate using water as vehicle. No increase in number of revertants observed indicating the substance to be non mutagenic with or without activation. Sodium fluorescein is non mutagenic in the Ames test performed for the Salmonella typhimurium TA1538, TA98, and TA100,TA 1537 and TA 1535.

 

In the study conducted by Mortelmans et al in 1986, Fluorescein, disodium salt (Acid yellow 73; CAS no 518 -47 -8) was examined for its ability to cause mutagenic changes when tested in five strains of the bacteria Salmonella typhimurium, specifically, TA 1535, TA 1537, TA 98 and TA 100 through the preincubation assay method. The test was conducted both in the presence and absence of metabolic activation using rat and hamster liver derived S-9 mix, over a range of doses, from 0 to 10000 ug/plate. Based on the results of this study, the test substanceFluorescein, disodium salt (Acid yellow 73) was not considered to be mutagenic under the conditions of this test.

 

Salmonella –Microsome assay was conducted (Toxicological and ecotoxicological assessment of water tracers, 2001) to study the genotoxicity of chemical Uranine (CAS no 518 -47 -8).The test chemical was to found to non – mutagenic in the Salmonella – Microsome assay. Fluorescent dyes that showed no effect upon either the genotoxicity or the ecotoxicity tests were classified by the Working Group as SAFE for use in water tracing. Uranine is classified as SAFE for use in water tracing.

 

From the same study, Chromosomal Aberration assay was conducted to study the genetoxicity of chemical Uranine (CAS no 518 -47 -8).The test chemical was to found to non – mutagenic in the Chromosomal Aberration assay using a mammalian cell culture. Fluorescent dyes that showed no effect upon either the genotoxicity or the ecotoxicity tests were classified by the Working Group as SAFE for use in water tracing. Uranine is classified as SAFE for use in water tracing.

 

Fluorescein, 2Na (CAS no 518 -47 -8) was tested (K.S. Loveday, B.E. Anderson, M.A. Resnick, and E. Zeiger, 1990) for its ability to induce sister chromosomal aberrations (ABs) in cultured Chinese hamster ovary (CHO) cells using a standard protocol with and without exogenous metabolic activation. The target substance however failed to induce chromosomal aberration with and without activation in the Chinese hamster ovary cell line and hemce is considered to be non-mutagenic.

 

In the study conducted by Katsuaki et al in 2004,Acid yellow 73was examined for its ability to cause mutagenic changes in Salmonella umu Test when tested in the bacteria Salmonella typhimurium, specifically,TA1535/pSK1002through the preincubation assay method. The test was conducted both in the presence and absence of metabolic activation using rat liver derived S-9 mix, over a range of doses, from 0 to 5000 ug/plate. Based on the results of this study, the test substanceFluorescein, disodium salt (Acid yellow 73) was not considered to be mutagenic under the conditions of this test

 

In the study conducted by Brian C. et al in 1991,Acid yellow 73was examined for its ability to cause mutagenic changes in L5178Y Mouse lymphoma cells in medium prepared for growing cultures. The test was conducted both in the presence and absence of metabolic activation using rat liver derived S-9 mix, over a range of doses, from 200 to 1000 ug/plate. Based on the results of this study, the test substanceFluorescein, disodium salt (Acid yellow 73) was considered to be non-mutagenic with metabolic activation system and mutagenic without metabolic activation under the conditions of this study.

 

In the study conducted by Nathan R. et al in 2003, Fluorescein, disodium salt (Acid yellow 73; CAS no 518 -47 -8) was examined for its ability to cause mutagenic changes when tested Chinese hamster lung (CHL) or Chinese hamster ovary (CHO) fibroblast cells.Compund is represented by calculated structural descriptors that encode topological, geometric, electronic, and polar surface features. A genetic algorithm (GA) employing a k-nearest neighbor (kNN) fitness evaluator is used to iteratively search a reduced descriptor space to find small, information-rich subsets of descriptors that maximize the classification rates for clastogenic and nonclastogenic responses. To further improve modeling, a similarity measure using atom-pair descriptors is employed to create more homogeneous data subsets. Negative responses were obtained in all three different data sets are examined. In all the three data sets Acid yellow 73 was negative. Hence, based on the results of this study, the test substanceFluorescein, disodium salt (Acid yellow 73) was not considered to be mutagenic under the conditions of this test.

 

D&C yellow No.8 (CAS no 518 -47 -8) was tested for mutagenic potential with the Salmonella/ mammalian -microsome test. In the plate incorporation assay performed (Jeanne M. Muzzall And Warren L. Cook, 1979), the 2 ml of liquid top agar was cooled to 45°C and 0.1 ml of a broth culture of microorganism and test substance in volumes of ≤0.4 ml of DMSO was added prior to placing on minimal agar plates. In all tests, the top agar was used with and without the S9 mix. After 48 h incubation at 37°C, the colonies which reverted to the prototroph were counted and compared to counts on the control plate to demonstrate mutagenicity or toxicity. Materials which caused a 2-fold increase of revertants, as compared to the number of spontaneous revertants on the control plates, were denoted as mutagens. Those which reduced the number of revertants were considered inhibitory. D&C yellow No.8 failed to induce an increase in the number of revertants and hence is non-mutagenic in nature.

Based on the information observed for the test chemical, it is summarized that Fluorescein sodium (CAS no 518 -47 -8) is not likely to exhibit genetic toxicity. Thus, the chemical is not classified as a genetic toxicant.


Justification for selection of genetic toxicity endpoint
Data is fom peer reveiw publication

Justification for classification or non-classification

Based on the information observed for the test chemical, it is summarized that Fluorescein sodium (CAS no 518 -47 -8) is not likely to exhibit genetic toxicity in vitro. Thus, the chemical is not classified as a genetic toxicant.