Nickel, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, chlorosulfonyl derivs., reaction products with 2-[(4-aminophenyl)sulfonyl]ethyl hydrogen sulfate monosodium salt, sodium salts

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:

1 November 1983 to 3 November 1983

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1983

Report date:

1983

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

See "Principles of method if other than guideline" below.

Deviations:

no

Principles of method if other than guideline:

AMES, B.N. et al. (1973) "Carcinogens are mutagens: a simple test system combining liver homogenates for activation and bacteria for detection." Proceedings of the National Academy of Sciences 70, 2281 - 2285.
GREEN, M.H.L. and ~URIEL, W.J. (1976) "Mutagen testing using TRP+ reversion of Escherichia coli.".Mutation Research 38, 3 - 32.

GLP compliance:

no

Remarks:

Study pre-dates GLP

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Nickel, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, chlorosulfonyl derivs., reaction products with 2-[(4-aminophenyl)sulfonyl]ethyl hydrogen sulfate monosodium salt, sodium salts

EC Number:

305-644-9

EC Name:

Nickel, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, chlorosulfonyl derivs., reaction products with 2-[(4-aminophenyl)sulfonyl]ethyl hydrogen sulfate monosodium salt, sodium salts

Cas Number:

94891-43-7

Molecular formula:

C32 H16 N8 Ni . C8 H11 N O6 S2 . 2 Na

IUPAC Name:

Nickel, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, chlorosulfonyl derivs., reaction products with 2-[(4-aminophenyl)sulfonyl]ethyl hydrogen sulfate monosodium salt, sodium salts

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

No further details specified in the study report.

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Additional strain / cell type characteristics:

not specified

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Additional strain / cell type characteristics:

not specified

Species / strain 3

Species / strain / cell type:

E. coli WP2 uvr A

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

4, 20, 100, 500, 2500 and 10000 μg/plate
No justification given for top dose.

Vehicle / solvent:

Not specified

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Aqua dest.

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

other: 2-Aminoanthracene; Methylhydrazone Derivative; Streptocotocine; ENNG

Details on test system and experimental conditions:

The tester strains of Salmonella typhimurium TA98, TA100, TA1535, TA1537, TA1538 and Escherichia coli WP2 uvrA were maintained in liquid nitrogen and recultivated for the test in fresh nutrient broth.
The S-9 fraction was prepared from the microsomes of a liver homogenate as follows. Sprague-Dawley rats were injected once with 500 mg/kg of polychlorinated biphenyl (PCB) = Aroclor 1254 U1onsanto, S. Louis, Mo., USA) in corn oil (200 mg/ml i.p.). Five days later the animals were killed, the livers removed and homogenised in three volumes of cold 0.15 M potassium chloride. The homogenate was centrifuged at 9,000g/10 min. One ml of supernatant then contains microsomes equivalent to about 250 mg liver. The preparation was stored in liquid nitrogen until required for use. Samples were thawed immediately before each test and the S-9 Mix is prepared.
To 2 ml of molten (45°C) top agar in a sterile test-tube were added 0.1 ml of the tester strain culture, graded quantities of the test compound in 0.1 ml solution and, for the S-9 series, 0.5 ml S-9 Mix. The contents of the test-tube were rapidly mixed and poured onto the surface of previously prepared minimal agar plates with VogelBonner E mixture.
The plates were incubated upside down at 37°C for 2 days, after which the number of revertant colonies appearing was counted.
Control plates without admixture of test compound were prepared in the same way to determine the spontaneous mutation rate.

Rationale for test conditions:

Not specfied

Evaluation criteria:

A chemical is considered to have a positive response if the number of induced revertants is more than double the spontaneous mutations.

Statistics:

Not specified

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 6

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

No unforeseen circumstances were observed which may have affected the quality or integrity of this study.

Any other information on results incl. tables

Results

μg/plate	S-9 Mix	Revertant colonies / plate TA 1537
10,000 2,500 500 100 20 4   10,000 2,500 500 100 20 4	- - - - - -   + + + + + +	3 – 6 – 7 – 8 2 – 3 – 5 – 6 3 – 4 – 5 – 7 3 – 4 – 6 – 6 3 – 5 – 6 – 8 4 – 4 – 6 – 7   4 – 6 – 9 – 10 7 – 8 – 13 – 14 9 – 12 – 14 – 16 9 – 10 – 12 – 13 11 – 11 – 13 – 18 13 – 13 – 16 – 17
Control (Aqua dest.) 0 0	- +	4 – 5 – 6 – 8 11 – 12 – 16 – 17
Positive controls 9-Aminoacridine 100 2-Aminoacridine 1 2-Aminoacridine 1	-   -   +	910 – 1080   7 – 9   90 – 99

 

μg/plate	S-9 Mix	Revertant colonies / plate TA 98
10,000 2,500 500 100 20 4   10,000 2,500 500 100 20 4	- - - - - -   + + + + + +	16 – 22 – 24 – 24 18 – 22 – 22 – 25 16 – 20 – 20 – 22 19 – 25 – 26 – 30 18 – 22 – 24 – 24 24 – 26 – 29 – 30   23 – 30 – 34 – 35 29 – 30 – 34 – 37 29 – 35 – 37 – 43 28 – 32 – 34 – 35 33 – 35 – 39 – 43 30 – 36 – 38 – 45
Control (Aqua dest.) 0 0	- +	17 – 21 – 23 – 24 39 – 41 – 46 – 47
Positive controls Methylhydrazone Derivative 5 2-Aminoacridine 0.5 2-Aminoacridine 0.5	-   -   +	3040 – 3480   25 – 25   445 – 490

 

μg/plate	S-9 Mix	Revertant colonies / plate TA 1538
10,000 2,500 500 100 20 4   10,000 2,500 500 100 20 4	- - - - - -   + + + + + +	3 – 5 – 6 – 8 4 – 5 – 7 – 11 4 – 5 – 5 – 6 5 – 6 – 6 – 7 6 – 7 – 7 – 9 7 – 9 – 9 – 10   7 – 7 – 8 – 10 7 – 9 – 12 – 13 12 – 12 – 16 – 17 13 – 16 – 18 – 20 15 – 18 – 18 – 19 13 – 14 – 17 – 19
Control (Aqua dest.) 0 0	- +	5 – 6 – 7 – 8 13 – 15 – 16 – 18
Positive controls Methylhydrazone Derivative 5 2-Aminoacridine 0.5 2-Aminoacridine 0.5	-   -   +	2060 – 2440   10 – 11   580 – 595

 

μg/plate	S-9 Mix	Revertant colonies / plate TA 100
10,000 2,500 500 100 20 4   10,000 2,500 500 100 20 4	- - - - - -   + + + + + +	105 – 117 – 126 – 139 107 – 111 – 117 – 128 118 – 141 – 143 – 143 129 – 130 – 131 – 146 128 – 133 – 140 – 153 113 – 129 – 133 – 137   140 – 141 – 149 – 153 150 – 165 – 169 – 184 141 – 151 – 157 – 172 135 – 137 – 142 – 148 133 – 143 – 155 – 157 140 – 152 – 154 – 159
Control (Aqua dest.) 0 0	- +	125 – 127 – 147 – 154 134 – 136 – 153 – 163
Positive controls Methylhydrazone Derivative 5 2-Aminoacridine 0.5 2-Aminoacridine 0.5	-   -   +	>5000 - >5000   113 – 119   570 – 630

 

μg/plate	S-9 Mix	Revertant colonies / plate TA 1535
10,000 2,500 500 100 20 4   10,000 2,500 500 100 20 4	- - - - - -   + + + + + +	12 – 14 – 15 – 17 10 – 12 – 17 – 19 11 – 13 – 17 – 18 9 – 11 – 13 – 17 11 – 14 – 15 – 17 10 – 11 – 14 – 17   8 – 11 – 13 – 14 11 – 11 – 14 – 14 14 – 15 – 18 – 21 12 – 13 – 16 – 18 10 – 10 – 14 – 17 11 – 14 – 17 – 19
Control (Aqua dest.) 0 0	- +	11 – 13 – 15 – 18 11 – 12 – 13 – 17
Positive controls Streptocotocine 5 2-Aminoacridine 1 2-Aminoacridine 1	-   -   +	>5000 - >5000   9 – 12   106 – 114

 

μg/plate	S-9 Mix	Revertant colonies / plate E.coli WP2 uvrA
10,000 2,500 500 100 20 4   10,000 2,500 500 100 20 4	- - - - - -   + + + + + +	59 – 60 – 64 – 71 36 – 36 – 37 – 44 25 – 26 – 27 – 31 29 – 30 – 32 – 33 20 – 24 – 28 – 29 22 – 23 – 25 – 30   54 – 62 – 63 – 70 32 – 40 – 41 – 45 37 – 41 – 44 – 46 34 – 36 – 39 – 41 37 – 38 – 39 – 42 32 – 35 – 37 – 40
Control (Aqua dest.) 0 0	- +	29 – 30 – 33 – 33 33 – 37 – 37 - 40
Positive controls ENNG 2 2-Aminoacridine 10 2-Aminoacridine 10	-   -   +	615 – 670   28 – 35   2380 – 2500

 

Applicant's summary and conclusion

Conclusions:

Under the conditions we employed Remazol-Brillantgrün 6 B in concentrations of 4 μg to 10,000 μg showed no mutagenic activity.

Executive summary:

Compound Remazol-Brillantgrün 6 B was examined in the mutagenicity screening test in bacteria first described by Ames and co-workers.

 

The Ames test is a well - established screening method for mutagenic activity. Five strains of Salmonella typhimurium (TA98, TA100, TA1535, TA1537 and TA1538), auxotrophic for histidine, and one of Escherichia coli (WP2 uvrA), auxotrophic for tryptophane, were used.

 

No unforeseen circumstances were observed which may have affected the quality or integrity of this study.

 

Under the conditions we employed Remazol-Brillantgrün 6 B in concentrations of 4 μg to 10,000 μg showed no mutagenic activity.