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Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

AMES test

The test chemical was considered  to be  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.

In vitro gene mutation assay in Mammalian cells

The test chemical was cosidered to be  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.

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
Data is from publication
Qualifier:
according to guideline
Guideline:
other: As mention below
Principles of method if other than guideline:
To determine the genetic toxicity of the test material by AMES test.
GLP compliance:
not specified
Type of assay:
bacterial gene mutation assay
Target gene:
AMES Assay
Species / strain / cell type:
S. typhimurium, other: No detailed data available
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Metabolic activation:
not specified
Metabolic activation system:
no data
Test concentrations with justification for top dose:
no data
Vehicle / solvent:
no data
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
not specified
True negative controls:
not specified
Positive controls:
not specified
Positive control substance:
not specified
Remarks:
no data
Details on test system and experimental conditions:
no data
Evaluation criteria:
An increase in the number of revertants was noted
Statistics:
no data
Species / strain:
S. typhimurium, other: No detailed data available
Metabolic activation:
not specified
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Remarks on result:
other: No mutagenic potential

Table 1 : Genetoxicity of the tracer dyes

Tracer

Salmonella –Microsome assay

Cytogenetic assay – Chromosal aberration

Pyranine

-

-

Conclusions:
The test chemical was considered to be 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.
Executive summary:

Uncertainties regarding possible negative effects on the environment or on human health of authorizing tracing experiments in groundwater and surface waters led to the establishment of a Working Group at the German Federal Environmental Agency (Umweltbundesamt – UBA) for conducting a toxicological and ecotoxicological assessment. A total of 17 water tracers was assessed by the Working Group on the basis of the results of toxicological tests. Two test methods that have been sufficiently validated and established for testing of chemicals were used: the salmonella/microsome test (gene mutation) and the analysis of chromosome aberration using a mammalian cell culture (chromosome aberration).The test chemical was considered to be 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.Test substance is classified as SAFE for use in water tracing.

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
Data is from publication.
Qualifier:
according to guideline
Guideline:
other: As mention below
Principles of method if other than guideline:
To determine the genetic toxicity of the test material .
GLP compliance:
not specified
Type of assay:
in vitro mammalian chromosome aberration test
Target gene:
no data
Species / strain / cell type:
mammalian cell line, other: No detailed data available
Details on mammalian cell type (if applicable):
No data
Additional strain / cell type characteristics:
not specified
Metabolic activation:
not specified
Metabolic activation system:
no data
Test concentrations with justification for top dose:
No data
Vehicle / solvent:
no data
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
not specified
True negative controls:
not specified
Positive controls:
not specified
Positive control substance:
not specified
Details on test system and experimental conditions:
no data
Evaluation criteria:
Chromosmal abberration,gaps were noted in cell line.
Statistics:
no data
Species / strain:
mammalian cell line, other:
Metabolic activation:
not specified
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Additional information on results:
No data
Remarks on result:
other: No mutagenic potential was observed

Table 1: Genetic Toxicity of tracer dyes

Tracer

Salmonella Microsome assay

Cytogenetic assay – Chromosal aberration

Pyranine

-

-

Conclusions:
The test chemical was cosidered to be 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.
Executive summary:

Uncertainties regarding possible negative effects on the environment or on human health of authorizing tracing experiments in groundwater and surface waters led to the establishment of a Working Group at the German Federal Environmental Agency (Umweltbundesamt – UBA) for conducting a toxicological and ecotoxicological assessment. A total of 17 water tracers was assessed by the Working Group on the basis of the results of toxicological tests. Two test methods that have been sufficiently validated and established for testing of chemicals were used: the salmonella/microsome test (gene mutation) and the analysis of chromosome aberration using a mammalian cell culture (chromosome aberration).The test chemical was considered to be 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.Test substance is classified as SAFE for use in water tracing.

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

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Gene mutation in vitro:

Data for the various test chemicals was reviewed to determine the mutagenic nature of Solvent Green 7; Pyranine; trisodium 8-hydroxypyrene-1,3,6-trisulfonate (6358-69-6). The studies are as mentioned below:

AMES test

Uncertainties regarding possible negative effects on the environment or on human health of authorizing tracing experiments in groundwater and surface waters led to the establishment of a Working Group at the German Federal Environmental Agency (Umweltbundesamt – UBA) for conducting a toxicological and ecotoxicological assessment. A total of 17 water tracers was assessed by the Working Group on the basis of the results of toxicological tests. Two test methods that have been sufficiently validated and established for testing of chemicals were used: the salmonella/microsome test (gene mutation) and the analysis of chromosome aberration using a mammalian cell culture (chromosome aberration).The test chemical was considered to be 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.Test substance is classified as SAFE for use in water tracing.

Supported by other studies.

Ames assay was performed to investigate the potential of the test chemical to induce gene mutations in comparison to negative control according to the plate incorporation test (Trial I) and the pre-incubation test (Trial II) using the Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102. The assay was performed in two independent experiments both with and without liver microsomal activation. Each concentration, including the negative, positive controls was tested in triplicate. Based on the pre-experiment results, the test item was tested with the following concentrations 0.0 (NC), 0.005, 0.016, 0.050, 0.158, 0.501 mg/plate for main study, both in the presence of metabolic activation (+S9) and in the absence of metabolic activation (-S9). No substantial increase in revertant colony numbers in any of the tester strains were observed following treatment with the test chemical at any dose level in both the confirmatory trials, neither in the presence nor in the absence of metabolic activation (S9 mix). In conclusion, it is stated that during the described mutagenicity test and under the experimental conditions reported, the test chemical did not induce gene mutations by base pair changes or frame shifts in the genome of the strains used.  

Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed using S. typhimurium strains TA98 and TA100 with and without S9 metabolic activation system. The test was performed as per the plate incorporation assay at dose level of 0 (negative control), 50 or 200µg/plate. The chemical was dissolved in water. The result was considered positive when a reproducible, dose-related, at least two-fold increase in the number of revertants over background was observed. Concurrent positive and negative control chemicals were also included in the study. The test chemical did not induce a doubling of revertant colonies over the negative control using S. typhimurium strains TA98 and TA100 in the presence and absence of S9 metabolic activation system and hence the chemical is not likely to classify as a gene mutant in vitro.

In vitro gene mutation assay in Mammalian cells

Uncertainties regarding possible negative effects on the environment or on human health of authorizing tracing experiments in groundwater and surface waters led to the establishment of a Working Group at the German Federal Environmental Agency (Umweltbundesamt – UBA) for conducting a toxicological and ecotoxicological assessment. A total of 17 water tracers was assessed by the Working Group on the basis of the results of toxicological tests. Two test methods that have been sufficiently validated and established for testing of chemicals were used: the salmonella/microsome test (gene mutation) and the analysis of chromosome aberration using a mammalian cell culture (chromosome aberration).The test chemical was considered to be 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.Test substance is classified as SAFE for use in water tracing.

Supported by other studies.

In vitro mammalian chromosome aberration test was performed to determine the mutagenic nature of the test chemical. The study was performed using CHO-WBL cells in the presence and absence of exogeneous metabolic activation system. The test chemical was dissolved in DMSO and used at dose levels upto 5 mg/mL. In the chromosome aberration assay without activation, cells were exposed to the test chemical for 8 hr. The test chemical was washed off, and the cells were treated with 0.1µg/ml Colcemid for 2-2.5 hr. With metabolic activation, the cells were exposed to the test chemical plus the metabolic activation mixture for 2 hr, washed, incubated for 8 hr, and then treated with Colcemid for 2-2.5 hr. A delayed harvest was used in the aberration assay in most instances when cell cycle delay was observed in the SCE assay. In these tests the cell growth period was extended to about 20 hr. Cells were harvested. Air-dried slides were coded and stained with Giemsa. One hundred to 200 cells from each of the three highest scorable doses were analyzed and the chromosomal aberrations were scored. The test chemical did not induce chromosomal aberrations when tested to toxicity. Precipitate was evident at doses of 250µg/ml and above. Based on the observations made, the test chemical did not induce chromosome aberrations in the CHO-WBL cells in the presence and absence of exogeneous metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

 

In vitro mammalian chromosome aberration test was performed to determine the mutagenic nature of the test chemical. The study was performed using CHO-WBL cells in the presence and absence of exogeneous metabolic activation system. The test chemical was dissolved in DMSO and used at dose levels upto 5 mg/mL. In the chromosome aberration assay without activation, cells were exposed to the test chemical for 8 hr. The test chemical was washed off, and the cells were treated with 0.1µg/ml Colcemid for 2-2.5 hr. With metabolic activation, the cells were exposed to the test chemical plus the metabolic activation mixture for 2 hr, washed, incubated for 8 hr, and then treated with Colcemid for 2-2.5 hr. A delayed harvest was used in the aberration assay in most instances when cell cycle delay was observed in the SCE assay. In these tests the cell growth period was extended to about 20 hr. Cells were harvested. Air-dried slides were coded and stained with Giemsa. One hundred to 200 cells from each of the three highest scorable doses were analyzed and the chromosomal aberrations were scored. The test chemical did not induce chromosomal aberrations when tested to toxicity. Precipitate was evident at doses of 250µg/ml and above. Based on the observations made, the test chemical did not induce chromosome aberrations in the CHO-WBL cells in the presence and absence of exogeneous metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

 

Based on the data summarized, Solvent Green 7; Pyranine; trisodium 8-hydroxypyrene-1,3,6-trisulfonate (6358-69-6)did not induce gene mutation .Hence it is not likely to be mutagenic in vitro.

Justification for classification or non-classification

Thus based on the above annotation and CLP criteria the test chemical Solvent Green 7; Pyranine; trisodium 8-hydroxypyrene-1,3,6-trisulfonate (6358-69-6) did not induce gene mutation .Hence it is not likely to be mutagenic in vitro.