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Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Pigment yellow 100 was studied for its ability to induce mutations in strains of Salmonella typhimurium.

 

The test compound was dissolved in DMSO and was tested at concentration of 0, 100, 333, 1000, 3333, 6666 or 10000 µg/plate using Salmonella typhimurium TA100, TA1535, TA1537, TA97 and TA98 in the presence and absence of 10 % and 30 % rat and hamster liver S9 metabolic activation system. Preincubation assay was performed with a preicubation for 20 mins. The plates were observed for histidine independence after 2 days incubation period. Concurrent solvent and positive controls were included in the study.

 

Pigment yellow 100 is not mutagenic to the Salmonella typhimurium TA100, TA1535, TA1537, TA97 and TA98 in the presence and absence of rat and hamster liver S9 metabolic activation system  and hence it is not likely to classify as a gene mutant in vitro.

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)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from peer reviewed publication
Qualifier:
according to
Guideline:
other: Refer below principle
Principles of method if other than guideline:
The test chemical Pigment yellow 100 (C.I. 19140) was studied for its ability to induce mutations in strains of Salmonella typhimurium.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay
Specific details on test material used for the study:
- Name of the test material: Pigment yellow 100- IUPAC name: aluminum tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl]diazenyl}benzenesulfonate) - Molecular formula: C48H33AlN12O27S6- Molecular weight: 495.4038 g/mol- Substance type: Organic- Purity: Labelled: 26%; Analyzed: 27%- Inchi: 1S/C16H12N4O9S2.Al/c21-15-13(18-17-9-1-5-11(6-2-9)30(24,25)26)14(16(22)23)19-20(15)10-3-7-12(8-4-10)31(27,28)29;/h1-8,13H,(H,22,23)(H,24,25,26)(H,27,28,29);/b18-17+;
Target gene:
Histidine
Species / strain / cell type:
other: TA100, TA1535, TA1537, TA97 and TA98
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
No data
Metabolic activation:
with and without
Metabolic activation system:
10% and 30% HLI and RLI S-9 (9,000 g supernatant) fractions were prepared from Aroclor 1254-induced, male Sprague- Dawley rat and male Syrian hamster livers
Test concentrations with justification for top dose:
0, 100, 333, 1000, 3333, 6666 or 10000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Dimethyl Sulfoxide (DMSO)- Justification for choice of solvent/vehicle: The test chemical was soluble in DMSO
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
For strains tested with S9
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive control substance:
sodium azide
Remarks:
For strains TA100 and TA1535 tested in the absence of S9
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
For strains TA97 and TA1537 tested in the absence of S9
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: 4-nitro-o-phenylenediamine
Remarks:
For strain TA98 tested in the absence of S9
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubationDURATION- Preincubation period: 20 mins- Exposure duration: 48 hrs- Expression time (cells in growth medium): 48 hrs- 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 availableSELECTION AGENT (mutation assays): No data availableSPINDLE INHIBITOR (cytogenetic assays): No data availableSTAIN (for cytogenetic assays): No data availableNUMBER OF REPLICATIONS: TriplicateNUMBER OF CELLS EVALUATED: No data availableDETERMINATION OF CYTOTOXICITY- Method: mitotic index; cloning efficiency; relative total growth; other: No data availableOTHER EXAMINATIONS:- Determination of polyploidy: No data available- Determination of endoreplication: No data available- Other: No data availableOTHER: No data available
Rationale for test conditions:
No data
Evaluation criteria:
Evaluations were made at both the individual trial and overall chemical levels.Individual trials were judged mutagenic (+), weakly mutagenic (+ W), questionable(?), or nonmutagenic (-), depending on the magnitude of the increase of his+ revertants, and the shape of the dose-response. A trial was considered questionable (?) if the dose-response was judged insufficiently high to support a call of “ +W,” if only a single dose was elevated over the control, or if the increase seen was not dose related. The distinctions between a questionable mutagenic response and a nonmutagenic or weak mutagenic response, and between a weak mutagenic response and mutagenic response are highly subjective. It was not necessary for a response to reach two fold over background for a chemical to be judged mutagenic.A chemical was judged mutagenic (+) or weakly mutagenic (+ W) if it produced a reproducible dose-related reponse over the solvent control in replicate trials. A chemical was judged questionable (?) if the results of individual trials were not reproducible, if increases in his+ revertants did not meet the criteria for a “+W” response, or if only single doses produced increases in his+ revertants in repeat trials.Chemicals were judged nonmutagenic (-) if they did not meet the criteria for a mutagenic or questionable response. The chemicals were decoded by the chemical repository only after a determination had been made regarding their mutagenicity or nonmutagenicity.
Statistics:
Mean ± SD
Species / strain:
other: TA100, TA1535, TA1537, TA97 and TA98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
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 availableRANGE-FINDING/SCREENING STUDIES: All chemicals were tested initially in a toxicity assay to determine the appropriate dose range for the mutagenicity assay. The toxicity assay was performed using TA100. Toxic concentrations were those that produced a decrease in the number of his+ colonies, or a clearing in the density of the background lawn, or both. COMPARISON WITH HISTORICAL CONTROL DATA: YesADDITIONAL INFORMATION ON CYTOTOXICITY: No data available
Remarks on result:
other: No mutagenic potential

Strain: TA100

Dose No Activation 
(Negative)
No Activation 
(Negative)
30% RLI 
(Negative)
30% HLI 
(Negative)
10% RLI 
(Negative)
10% HLI 
(Negative)
Protocol Preincubation Preincubation Preincubation Preincubation Preincubation Preincubation
ug/Plate Mean ±SEM Mean ±SEM Mean ±SEM Mean ±SEM Mean ±SEM Mean ±SEM
 0         
146 10 127 7.5 145 14.7 167 6.8 107 10.1 107 6.8
100         
133 3.7 137 7.5 143 4.7 165 17.8 123 16.5 117 11.4
333         
130 3.8 152 5 116 5 155 10.8 122 4.4 128 8.7
1000         
146 6.1 136 6.7 127 .7 116 16.5 130 13.5 117 13.1
3333         
109 4 65 7.9 130 15.3 118 5.9 106 5.8 85 8.7
6666         
16 .7 75 11.4 53 8.9
10000         
37 9 65 8.2 61 12.6
Positive Control 604 4 550 21.1 478 14.7 529 14.2 515 46 548 52.7

Strain: TA1535

Dose No Activation 
(Negative)
No Activation 
(Negative)
30% RLI 
(Negative)
30% HLI 
(Negative)
10% RLI 
(Negative)
10% HLI 
(Negative)
Protocol Preincubation Preincubation Preincubation Preincubation Preincubation Preincubation
ug/Plate Mean ±SEM Mean ±SEM Mean ±SEM Mean ±SEM Mean ±SEM Mean ±SEM
 0         
31 1 20 .6 15 1.8 9 2 8 .6 10 1.5
100         
26 1.5 20 1.5 14 1.9 10 1.8 11 1.2 12 2
333         
30 5.2 16 .6 16 2.2 8 1.2 7 2.5 7 .9
1000         
30 4 25 6.9 15 1.9 8 1.5 9 .9 8 .9
3333         
24 3.7 15 2.5 11 1.2 8 3.1 9 2.7 6 1.5
6666         
14 2.6 6 .9 6 .3
10000         
21 1.9 13 1.9 7 .7
Positive Control 357 22.3 499 21.5 218 6.6 388 17.1 121 8.4 219 4.6

Strain: TA1537

Dose No Activation 
(Negative)
30% RLI 
(Negative)
30% HLI 
(Negative)
Protocol Preincubation Preincubation Preincubation
ug/Plate Mean ±SEM Mean ±SEM Mean ±SEM
 0         
8 .7 14 1.5 15 .6
100         
6 1.5 11 1.7 12 3.5
333         
10 1.9 15 1.8 12 1.9
1000         
11 1.8 12 .6 10 .3
3333         
6 .6 9 1.5 6 0
10000         
5 1.2 8 1.9 4 0
Positive Control 309 5 41 2.3 46 3.3
Conclusions:
Pigment yellow 100 is not mutagenic to the Salmonella typhimurium TA100, TA1535, TA1537, TA97 and TA98 in the presence and absence of rat and hamster liver S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.
Executive summary:

Pigment yellow 100 was studied for its ability to induce mutations in strains of Salmonella typhimurium.

 

The test compound was dissolved in DMSO and was tested at concentration of 0, 100, 333, 1000, 3333, 6666 or 10000 µg/plate using Salmonella typhimurium TA100, TA1535, TA1537, TA97 and TA98 in the presence and absence of 10 % and 30 % rat and hamster liver S9 metabolic activation system. Preincubation assay was performed with a preicubation for 20 mins. The plates were observed for histidine independence after 2 days incubation period. Concurrent solvent and positive controls were included in the study.

 

Pigment yellow 100 is not mutagenic to the Salmonella typhimurium TA100, TA1535, TA1537, TA97 and TA98 in the presence and absence of rat and hamster liver S9 metabolic activation system  and hence it is not likely to classify as a gene mutant in vitro.

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 available for the target chemical and its reas across have been reviewed to determine the mutagenic nature of aluminum tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl]diazenyl}benzenesulfonate). The studies are as mentioned below:

Pigment yellow 100 (CAS no 12225 -21 -7) was studied by Zeiger et al (Environmental and Molecular Mutagenesis, 1998) for its ability to induce mutations in strains of Salmonella typhimurium. The test compound was dissolved in DMSO and was tested at concentration of 0, 100, 333, 1000, 3333, 6666 or 10000 µg/plate using Salmonella typhimurium TA100, TA1535, TA1537, TA97 and TA98 in the presence and absence of 10 % and 30 % rat and hamster liver S9 metabolic activation system. Preincubation assay was performed with a preicubation for 20 mins. The plates were observed for histidine independence after 2 days incubation period. Concurrent solvent and positive controls were included in the study. Pigment yellow 100 is not mutagenic to the Salmonella typhimurium TA100, TA1535, TA1537, TA97 and TA98 in the presence and absence of rat and hamster liver S9 metabolic activation system  and hence it is not likely to classify as a gene mutant in vitro.

The above data for the target chemical is further supported by the data from read across chemical.

In a study performed by Das and Mukherjee (Human Genetics, 2004) Ames mutagenicity assay was performed to evaluate the mutagenic nature of the structurally and functionally similar read across chemical tartrazine (RA CAS no 1934 -21 -0; IUPAC name: Trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo)pyrazole-3-carboxylate) using the plate incorporation assay. Tartrazine was dissolved in sterile double distilled water and was tested at a concentration of 0, 100, 250, 500 and 1000 μg /plate. The plates were inverted within an hour and placed in a dark vented incubator at 37⁰C for 48 hours. Positive controls (for TA97a and TA98, 20 μg/plate nitro phenylene diamine and for TA100, 1.5 μg/plate sodium azide) and negative controls were maintained concurrently for all the experiments. Three plates were used for each set. After 48 hours of incubation, the revertant colonies were counted. Tartrazine did not induce gene mutation in Salmonella typhimurium strains TA97a, TA98 and TA100 and hence it is not likely to classify as a gene mutant in vitro.

Chromosomal aberration test was performed by Ishidate et al (Food and chemical toxicology, 1988) for the structurally and functionally similar read across chemical tartrazine (RA CAS no 1934 -21 -0; IUPAC name:Trisodium 5-hydroxy-1-(4-sulphophenyl)-4-(4-sulphophenylazo)pyrazole-3-carboxylate) using Chinese hamster fibroblast cell line CHL. The cells were exposed to the test material at three different doses with 2.5 mg/plate being the highest dose for 24 and 48 hr. In the present studies, no metabolic activation systems were applied. The incidence of polyploid cells as well as of cells with structural chromosomal aberrations such as chromatid or chromosome gaps, breaks, exchanges, ring formations, fragmentations and others, was recorded on each culture plate. Untreated cells and solvent-treated cells served as negative controls, in which the incidence of aberrations was usually less than 3.0%. The results were considered to be negative if the incidence was less than 4.9%, equivocal if it was between 5.0 and 9.9%, and positive if it was more than 10.0%. The incidence of chromosome aberration in Chinese hamster fibroblast cell line for the test chemical tartrazine was considered to be more than 10% in the absence of metabolic activation system during the 48 hrs study duration and hence tartrazine is mutagenic in vitro.

In the same study by Ishidate et al, Gene mutation toxicity study was performed to determine the mutagenic nature of tartrazine (RA CAS no 1934 -21 -0). The study was performed using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 with and without S9 metabolic activation system. The test was performed as per the preincubation assay at six different concentrations with 5.0 mg/plate being the maximum concentration. The chemical was dissolved in distilled water. Preincubation was performed for 20 mins and the exposure duration was for 48 hrs. The result was considered positive if the number of colonies found was twice the number in the control (exposed to the appropriate solvent or untreated). Tartrazine did not induce a doubling of revertant colonies over the control using S. typhimurium strains TA92, TA1535, TA100, TA1537, TA94 and TA98 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.

Ivett et al (Environmental and Molecular Mutagenesis, 1989) performed chromosomal aberration study for another strcturally and functionally simlar read across chemical. Chromosomal aberration tests were carried out to determine the mutagenic nature of FD and C yellow 6 (RA CAS no 2783 -94 -0; IUPAC name: disodium (5E)-6-oxo-5-[(4-sulfonatophenyl)hydrazinylidene]naphthalene-2-sulfonate). The study was performed using Chinese hamster ovary cells in the presence and absence of S9 metabolic activation system. FD and C yellow no 6 was dissolved in medium and used at dose levels of up to 5 mg/mL. In the assays for chromosomal aberrations, the top dose (TD) was based on toxicity, solubility, or the upper testing limit (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 0.1 µg/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. Air-dried slides were coded and stained with Giemsa. One hundred to 200 cells from each of the three highest scorable doses were analyzed. All aberrations were individually classified (e.g., chromatid breaks, chromosome breaks, triradials, etc.). These data were combined as the percent of cells with simple (deletions), complex (exchanges), and total (simple, complex and other) aberrations. FD and C yellow no 6 did not induce chromosome aberrations in Chinese hamster ovary cells (CHO) in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

Gene mutation toxicity study was also performed by CHung et al (Applied and Environmental Microbiology, 1981) to determine the mutagenic nature of Sunset yellow (RA CAS no 2783 -94 -0, IUPAC name: disodium (5E)-6-oxo-5-[(4-sulfonatophenyl)hydrazinylidene]naphthalene-2-sulfonate). The study was performed by the standard plate incorporation assay using Salmonella typhimurium strains TA1535, TA1537, TA1538, TA98, and TA100 with and without S9 metabolic activation system. The test chemical was dissolved in DMSO and upto a maximum nontoxic dose of 5000 µg/plate. Concurrent solvent and positive controls were also included in the study. Sunset yellow did not induce gene mutation in Salmonella typhimurium strains TA1535, TA1537, TA1538, TA98, and TA100 in the presence and absence of S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

Based on the data available for the target chemical and its read across, aluminum tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl]diazenyl}benzenesulfonate) does not exhibit gene mutation in vitro. Hence it is not likely to classify as a gene mutant as per the criteria mentioned in CLP regulation. The read across chemical with CAS no 1934 -21 -0 is the parent compound for aluminum tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl]diazenyl}benzenesulfonate) and its depicts the non mutagenic nature in the Ames test and the chromosomal aberration study performed with metabolic activation system. Considering this and the other read across data, the test chemical aluminum tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl]diazenyl}benzenesulfonate) is considered to be non-mutagenic in vitro.

Justification for classification or non-classification

Based on the data available for the target chemical and its read across, aluminum tris(4-{[3-carboxy-5-oxo-1-(4-sulfophenyl)-4,5-dihydro-1H-pyrazol-4-yl] diazenyl}benzenesulfonate) (CAS no 12225 -21 -7) does not exhibit gene mutation in vitro. Hence it is not likely to classify as a gene mutant as per the criteria mentioned in CLP regulation.