Copper, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, chlorosulfonyl derivs., reaction products with 2-[(4-aminophenyl)amino]-5-nitrobenzenesulfonic acid, 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:

From October 24, 2016 to December 20, 2016

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

other company data

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Copper, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, chlorosulfonyl derivs., reaction products with 2-[(4-aminophenyl)amino]-5-nitrobenzenesulfonic acid, sodium salts

EC Number:

277-403-5

EC Name:

Copper, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, chlorosulfonyl derivs., reaction products with 2-[(4-aminophenyl)amino]-5-nitrobenzenesulfonic acid, sodium salts

Cas Number:

73378-63-9

Molecular formula:

C32H16CuN8.C12H11N3O5S.Na

IUPAC Name:

Copper, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, chlorosulfonyl derivs., reaction products with 2-[(4-aminophenyl)amino]-5-nitrobenzenesulfonic acid, sodium salts

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): Everlan SL53
- Substance type: Powder
- Composition of test material, percentage of components: 73.75 %
- Lot/batch No.: 4508
- Storage condition of test material: Ambient

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

S9 Mix

Controls

Untreated negative controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

9-aminoacridine

sodium azide

benzo(a)pyrene

mitomycin C

other: 2-Aminoanthracene

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Untreated negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Table 1. Characteristics of five Salmonella typhimurium strains

Test strain	Histidine requirement	uvrB mutation	rfa mutation	Ampicillin resistance
TA98	+	+	+	+
TA100	+	+	+	+
TA102	+	ϴ	+	+
TA1535	+	+	+	ϴ
TA1537	+	+	+	ϴ

+ means had the characteristic; ϴ means did not have the characteristic

Table 2. Toxicity of the test article of Salmonella typhimurium TA100

Group	Test article (mg/plate)	Reverse mutant colony number (CFU/plate)
With S9 Mix	Negative control group	202
	Positive control group^a	1061
	Test group
	5	215
	2.5	157
	1.25	163
	0.625	167
	0.3125	199
Without S9 Mix	Negative control group	233
	Positive control group	686
	Test group
	5	178
	2.5	162
	1.25	174
	0.625	164
	0.3125	158

^a Positive control group: With S9 Mix: 2-Aminoanthracene (4.0μg/plate).

Without S9 Mix: Sodium azide (5μg/plate).

 

Table 3. Reverse mutation test of Salmonella typhimurium TA98

Group	Test article (mg/plate)	Reverse mutant colony number (CFU/plate)			Average of colonies^a
		1	2	3
With S9 Mix	Negative control group	39	39	36	38 ± 2
	Positive control group^b	225	274	261	253 ± 25*
	Test group
	5	27	20	22	23 ± 4
	2.5	25	21	35	27 ± 7
	1.25	47	33	41	40 ± 7
	0.625	25	33	32	30 ± 4
	0.3125	39	40	36	38 ± 2
Without S9 Mix	Negative control group	41	36	57	45 ± 11
	Positive control group	328	331	380	346 ± 29*
	Test group
	5	26	32	30	29 ± 3
	2.5	24	42	35	34 ± 9
	1.25	28	24	41	31 ± 9
	0.625	40	39	50	43 ± 6
	0.3125	60	54	43	52 ± 9

^a Average of colonies was shown as Mean ± S.D., the data were triplicate.

^b Positive control group: With S9 Mix: Benzo [a] pyrene (4.0μg/plate).

Without S9 Mix: 4-Nitroquinoline-N-oxide (0.5μg/plate).

^*Reverse mutant colony number were twice more than negative control group, ρ < 0.05 (One-Way ANOVA).

 

Table 4. Reverse mutation test of Salmonella typhimurium TA100

Group	Test article (mg/plate)	Reverse mutant colony number (CFU/plate)			Average of colonies^a
		1	2	3
With S9 Mix	Negative control group	221	211	285	239 ± 40
	Positive control group^b	1668	1712	1685	1688 ± 22
	Test group
	5	154	157	148	153 ± 5
	2.5	168	198	165	177 ± 18
	1.25	155	144	143	147 ± 7
	0.625	176	172	144	164 ± 17
	0.3125	182	187	178	182 ± 5
Without S9 Mix	Negative control group	186	171	196	184 ± 13
	Positive control group	730	725	718	724 ± 6*
	Test group
	5	151	136	124	137 ± 14
	2.5	169	198	183	183 ± 15
	1.25	177	152	148	159 ± 16
	0.625	169	133	156	153 ± 18
	0.3125	160	185	196	180 ± 18

^a Average of colonies was shown as Mean ± S.D., the data were triplicate.

^b Positive control group: With S9 Mix: 2-Aminoanthracene (4.0μg/plate).

Without S9 Mix: Sodium azide (5μg/plate).

^*Reverse mutant colony number were twice more than negative control group, ρ < 0.05 (One-Way ANOVA).

 

Table 5. Reverse mutation test of Salmonella typhimurium TA102

Group	Test article (mg/plate)	Reverse mutant colony number (CFU/plate)			Average of colonies^a
		1	2	3
With S9 Mix	Negative control group	423	412	438	424 ± 13
	Positive control group^b	900	901	998	933 ± 56*
	Test group
	5	356	364	423	381 ± 37
	2.5	427	435	425	429 ± 5
	1.25	426	466	382	425 ± 42
	0.625	440	409	445	431 ± 20
	0.3125	427	518	461	469 ± 46
Without S9 Mix	Negative control group	383	430	388	400 ± 26
	Positive control group	882	1046	1138	1022 ± 130*
	Test group
	5	311	336	322	323 ± 13
	2.5	382	351	369	367 ± 16
	1.25	310	366	343	340 ± 28
	0.625	349	334	343	342 ± 8
	0.3125	365	299	358	341 ± 36

^a Average of colonies was shown as Mean ± S.D., the data were triplicate.

^b Positive control group: With S9 Mix: 2-Aminoanthracene (10μg/plate).

Without S9 Mix: Mitomycin C (0.5μg/plate).

^*Reverse mutant colony number were twice more than negative control group, ρ < 0.05 (One-Way ANOVA).

 

Table 6. Reverse mutation test of Salmonella typhimurium TA1535

Group	Test article (mg/plate)	Reverse mutant colony number (CFU/plate)			Average of colonies^a
		1	2	3
With S9 Mix	Negative control group	28	30	31	30 ± 2
	Positive control group^b	381	390	331	367 ± 32*
	Test group
	5	27	28	26	27 ± 1
	2.5	29	31	25	28 ± 3
	1.25	33	32	28	31 ± 3
	0.625	28	29	17	25 ± 7
	0.3125	32	23	32	29 ± 5
Without S9 Mix	Negative control group	23	36	31	30 ± 7
	Positive control group	284	295	270	283 ± 13*
	Test group
	5	25	31	27	28 ± 3
	2.5	36	32	32	33 ± 2
	1.25	28	35	33	32 ± 4
	0.625	25	27	27	26 ± 1
	0.3125	25	29	36	30 ± 6

^a Average of colonies was shown as Mean ± S.D., the data were triplicate.

^b Positive control group: With S9 Mix: 2-Aminoanthracene (4.0μg/plate).

Without S9 Mix: Sodium azide (0.4μg/plate).

^*Reverse mutant colony number were twice more than negative control group, ρ < 0.05 (One-Way ANOVA).

 

Table 7. Reverse mutation test of Salmonella typhimurium TA1537

Group	Test article (mg/plate)	Reverse mutant colony number (CFU/plate)			Average of colonies^a
		1	2	3
With S9 Mix	Negative control group	20	13	15	16 ± 4
	Positive control group^b	342	320	331	331 ± 11*
	Test group
	5	13	15	11	13 ± 2
	2.5	17	14	14	15 ± 2
	1.25	13	17	18	16 ± 3
	0.625	16	14	18	16 ± 2
	0.3125	19	15	20	18 ± 3
Without S9 Mix	Negative control group	15	20	15	17 ± 3
	Positive control group	838	853	878	856 ± 20*
	Test group
	5	12	17	12	14 ± 3
	2.5	11	13	20	15 ± 5
	1.25	17	17	15	16 ± 1
	0.625	13	19	20	17 ± 4
	0.3125	15	15	20	17 ± 3

^a Average of colonies was shown as Mean ± S.D., the data were triplicate.

^b Positive control group: With S9 Mix: 2-Aminoanthracene (4.0μg/plate).

Without S9 Mix: 9-Aminoacridine (50.0μg/plate).

^*Reverse mutant colony number were twice more than negative control group, ρ < 0.05 (One-Way ANOVA).

Applicant's summary and conclusion

Conclusions:

According to OECD 471 test method, Everlan SL53 was not mutagenic in the reverse mutation analysis of Salmonella typhimurium up to 5 mg/plate.

Executive summary:

This test using the procedures outlined in the SuperLub Study Plan for M62-151100153001EN which is based on the SOP for the OECD 471 (SOPF-203) and OECD 471 (OECD, 1997). The results of this OECD 471 test for Everlan SL53 show that test validity criteria was met.

Based on the preliminary assay results, 5mg/platewas set as the highest dose in this study. In the mutagenicity assay, five doses of Everlan SL53 at 0.3125, 0.625, 1.25, 2.5 and 5mg/plate, concurrent negative and strain-specific positive controls were tested in tester strains TA98, TA100, TA102, TA1535 and TA1537 in triplicate with or without S9 Mix activation. No cytotoxicity was observed in all five tester strains up to 5mg/plate in the absence and presence of metabolite activations. Results showed that Everlan SL53 did not increase the number of revertants in all five tester strains TA98, TA100, TA102, TA1535 and TA1537 up to 5mg/plate either in the absence or in the presence of metabolite activation.

Based on the data obtained from this study, it was concluded that under the test condition, Everlan SL53 was not mutagenic in the reverse mutation analysis of Salmonella typhimurium up to 5mg/plate in the absence and presence of S9 metabolic activation.