Hydrogen [29H,31H-phthalocyaninesulphonato(3-)-N29,N30,N31,N32]cuprate(1-)

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The substance is non mutagenic in the Ames test (OECD 471, GLP).

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

Study period:

2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP and OECD guideline compliant study

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

(21 Jul 1997)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

(BASF SE, Experimental Toxicology and Ecology, Ludwigshafen, Germany)

Type of assay:

bacterial reverse mutation assay

Target gene:

his+ / trp+

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

phenobarbital and beta-naphthoflavone induced rat liver S9 microsomal fraction

Test concentrations with justification for top dose:

1st Experiment (Standard plate test with and without S9 mix): 0; 10; 33; 100; 333; 1000 and 1500 μg/plate
2nd Experiment (Preincubation test with and without S9 mix): 0; 10; 33; 100; 333; 1000 and 1500 μg/plate
3rd Experiment for TA 1535 (0; 10; 33; 100; 333; 1000 and 1500 μg/plate (with and without S9 mix) - performed because of contamination in 2nd experiment.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Due to the limited solubility of the test substance in ultrapure water, DMSO was used as vehicle, which had been demonstrated to be suitable in bacterial reverse mutation tests and for which historical control data are available.

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene (2-AA), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), 4-nitro-o-phenylenediamine (NOPD), 9-aminoacridine (AAC), 4-nitroquinoline-N-oxide (4-NQO)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); preincubation

Standard plate test:
- Exposure duration: 48 – 72 hours

Preincubation test:
- Preincubation period: 20 min
- Exposure duration: 48 – 72 hours


NUMBER OF REPLICATIONS: 3


DETERMINATION OF CYTOTOXICITY
- Method: decrease in the number of revertants, clearing or diminution of the background lawn, reduction in the titer


OTHER:
Titer determination: The titer was determined only in the experimental parts with S9 mix both for the negative controls (vehicle only) and for the two highest doses in all experiments.

Positive controls:

With S9 mix: 2-aminoanthracene (2-AA), 2.5 μg/plate, dissolved in DMSO / TA 1535, TA 100, TA 1537, TA 98; 60 μg/plate, dissolved in DMSO / Escherichia coli WP2 uvrA
Without S9 mix: N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), 5 μg/plate, dissolved in DMSO / TA 1535, TA 100; 4-nitro-o-phenylenediamine (NOPD), 10 μg/plate, dissolved in DMSO / TA 98; 9-aminoacridine (AAC), 100 μg/plate, dissolved in DMSO / TA 1537; 4-nitroquinoline-N-oxide (4-NQO) (SIGMA, N-8141), 5 μg/plate, dissolved in DMSO / E. coli WP2 uvrA

Evaluation criteria:

Acceptance criteria:
Generally, the experiment is considered valid if the following criteria are met:
• The number of revertant colonies in the negative controls was within the range of the historical negative control data for each tester strain.
• The sterility controls revealed no indication of bacterial contamination.
• The positive control substances both with and without S9 mix induced a distinct increase in the number of revertant colonies within the range of the historical positive control data or above.
• Fresh bacterial culture containing approximately 10^9 cells per mL were used. For approval the titer of viable bacteria was ≥ 10^8 colonies per mL.

Assessment criteria:
The test substance is considered positive in this assay if the following criteria are met:
• A dose-related and reproducible increase in the number of revertant colonies, i.e. about doubling of the spontaneous mutation rate in at least one tester strain either without S9 mix or after adding a metabolizing system.

A test substance is generally considered non-mutagenic in this test if:
• The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in at least two experiments carried out independently of each other.

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Test substance precipitation was found from about 333 μg/plate onward with and without S9 mix.

COMPARISON WITH HISTORICAL CONTROL DATA: yes

ADDITIONAL INFORMATION ON CYTOTOXICITY: A weak bacteriotoxic effect (slight decrease in the number of his+ revertants) was occasionally observed in the standard plate test depending on the strain and test conditions from about 1000 μg/plate onward.
In the preincubation assay weak bacteriotoxicity (slight decrease in the number of his+ revertants) was observed only in tester strains TA 1535 (without S9 mix) and TA 98 (with S9 mix) at a concentration of 1500 μg/plate.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Table 1: Standard plate test, with metabolic activation

strain		test group	Dose (μg/plate)		Mean revertants per plate	SD	Factor	Individual revertant colony counts
TA 1535		DMSO	-		12.3	4.2	-	11, 17, 9
		Test item	10		14.0	5.3	1.1	18, 16, 8
			33		10.0	2.6	0.8	7, 12, 11
			100		11.0	3.0	0.9	14, 11, 8
			333		8.7	0.6	0.7	9 P, 8 P, 9 P
			1000		7.7	2.1	0.6	6 P, 10 P, 7 P
			1500		7.3	2.1	0.6	9 P, 8 P, 5 P
		2-AA	2.5		326.7	17.5	26.5	346, 322, 312
TA 100		DMSO	-		61.3	10.3	-	64, 50, 70
		Test item	10		55.3	9.9	0.9	62, 44, 60
			33		57.0	4.0	0.9	53, 61, 57
			100		63.0	12.2	1.0	57, 55, 77
			333		53.0	10.4	0.9	65 P, 47 P, 47 P
			1000		49.0	5.6	0.8	54 P, 50 P, 43 P
			1500		60.0	9.0	1.0	60 P, 51 P, 69 P
		2-AA	2.5		2232.3	261.2	36.4	2509, 2198, 1990
TA 1537		DMSO	-		8.3	4.5	-	13, 8, 4
		Test item	10		9.7	1.5	1.2	11, 8, 10
			33		9.0	2.0	1.1	9, 11, 7
			100		8.7	2.1	1.0	7, 8, 11
			333		10.0	4.6	1.2	11 P, 14 P, 5 P
			1000		7.7	1.5	0.9	9 P, 8 P, 6 P
			1500		8.7	1.5	1.0	7 P, 10 P, 9 P
		2-AA	2.5		219.0	10.6	26.3	223, 227, 207
TA 98		DMSO	-		31.0	9.5	-	42, 26, 25
		Test item	10		27.3	5.7	0.9	29, 32, 21
			33		25.3	3.8	0.8	21, 28, 27
			100		22.7	1.2	0.7	22, 22, 24
			333		25.0	1.0	0.8	25 P, 26 P, 24 P
			1000		20.7	4.0	0.7	23 P, 16 P, 23 P
			1500		12.3	1.2	0.4	11 P, 13 P, 13 P
		2-AA	2.5		1871.7	467.4	60.4	2397, 1716, 1502
E. coli	DMSO			-	62.7	11.7	-	65, 50, 73
	Test item			10	72.0	6.6	1.1	71, 66, 79
				33	70.7	2.3	1.1	72, 72, 68
				100	77.3	1.5	1.2	76, 79, 77
				333	68.0	8.7	1.1	78 P, 63 P, 63 P
				1000	66.7	11.1	1.1	55 P, 77 P, 68 P
				1500	69.3	8.6	1.1	60 P, 71 P, 77 P
	2-AA			60	321.3	15.0	5.1	307, 337, 320

P = precipitation

Table 2: Standard incubation test, without metabolic activation

strain		test group	Dose (μg/plate)		Mean revertants per plate	SD	Factor	Individual revertant colony counts
TA 1535		DMSO	-		11.7	3.8	-	16, 10, 9
		Test item	10		14.3	5.5	1.2	17, 18, 8
			33		10.7	6.4	0.9	8, 18, 6
			100		7.7	2.5	0.7	8, 10, 5
			333		8.7	1.2	0.7	10 P, 8 P, 8 P
			1000		6.3	1.5	0.5	6 P, 5 P, 8 P
			1500		7.7	2.1	0.7	7 P, 10 P, 6 P
		MNNG	5.0		6504.3	118.3	557.5	6528, 6376, 6609
TA 100		DMSO	-		45.3	10.7	-	36, 43, 57
		Test item	10		36.7	9.8	0.8	48, 31, 31
			33		45.0	4.4	1.0	40, 47, 48
			100		42.3	6.7	0.9	44, 48, 35
			333		45.7	8.7	1.0	36 P, 48 P, 53 P
			1000		40.7	10.4	0.9	44 P, 49 P, 29 P
			1500		29.0	6.0	0.6	29 P, 23 P, 35 P
		MNNG	5.0		5768.7	118.1	127.3	5791, 5874, 5641
TA 1537		DMSO	-		6.7	1.2	-	6, 6, 8
		Test item	10		6.0	2.0	0.9	6, 8, 4
			33		5.3	1.2	0.8	6, 4, 6
			100		6.7	2.5	1.0	9, 7, 4
			333		6.3	0.6	1.0	6 P, 7 P, 6 P
			1000		3.0	1.0	0.5	3 P, 4 P, 2 P
			1500		5.3	0.6	0.8	6 P, 5 P, 5 P
		AAC	100		1966.3	109.8	295.0	1992, 1846, 2061
TA 98		DMSO	-		18.3	1.2	-	19, 19, 17
		Test item	10		16.7	6.4	0.9	13, 13, 24
			33		13.7	4.2	0.7	15, 9, 17
			100		17.3	1.5	0.9	17, 16, 19
			333		13.7	3.8	0.7	18 P, 12 P, 11 P
			1000		14.3	3.8	0.8	10 P, 17 P, 16 P
			1500		10.3	3.5	0.6	14 P, 10 P, 7 P
		NOPD	10		315.7	20.4	17.2	338, 311, 298
E. coli	DMSO		-		58.3	15.5	-	71, 63, 41
	Test item		10		59.7	5.7	1.0	66, 58, 55
			33		57.0	2.6	1.0	60, 55, 56
			100		52.7	4.0	0.9	57, 52, 49
			333		56.7	4.7	1.0	62 P, 53 P, 55 P
			1000		57.7	9.3	1.0	68 P, 55 P, 50 P
			1500		59.3	9.3	1.0	70 P, 53 P, 55 P
	4-NQO		5		776.7	10.0	13.3	773, 769, 788

P = precipitation

Table 3: Preincubation assay, without metabolic activation

strain		test group	Dose (μg/plate)		Mean revertants per plate	SD	Factor	Individual revertant colony counts
TA 1535		DMSO Test item	- 10		- -	- -	- -		- C, - C, - C - C, - C, - C
			33		-	-	-		- C, - C, - C
			100		-	-	-		- C, - C, - C
			333		-	-	-		- C P, - C P, - C P
			1000		-	-	-		- C P, - C P, - C P
			1500		-	-	-		- C P, - C P, - C P
		MNNG	5.0		2535.3	149.1	-		2390, 2528, 2688
TA 100		DMSO	-		42.7	9.0	-		52, 34, 42
		Test item	10		46.3	5.8	1.1		43, 53, 43
			33		44.0	11.8	1.0		31, 47, 54
			100		47.3	9.6	1.1		49, 56, 37
			333		42.0	11.3	1.0		48 P, 49 P, 29 P
			1000		39.3	5.1	0.9		38 P, 45 P, 35 P
			1500		34.3	3.5	0.8		31 P, 34 P, 38 P
		MNNG	5.0		2249.3	166.8	52.7		2364, 2326, 2058
TA 1537		DMSO	-		6.3	2.1	-		8, 7, 4
		Test item	10		7.7	0.6	1.2		7, 8, 8
			33		6.7	1.2	1.1		6, 8, 6
			100		6.7	1.5	1.1		7, 5, 8
			333		7.0	2.6	1.1		6 P, 5 P, 10 P
			1000		7.0	3.0	1.1		7 P, 4 P, 10 P
			1500		8.3	5.0	1.3		13 P, 9 P, 3 P
		AAC	100		1130.0	66.6	178.4		1195, 1133, 1062
TA 98		DMSO	-		21.3	9.6	-		30, 11, 23
		Test item	10		19.7	5.9	0.9		24, 22, 13
			33		17.3	0.6	0.8		17, 18, 17
			100		22.3	3.5	1.0		22, 19, 26
			333		21.7	7.2	1.0		17 P, 30 P, 18 P
			1000		19.0	1.7	0.9		17 P, 20 P, 20 P
			1500		14.3	1.5	0.7		13 P, 14 P, 16 P
		NOPD	10		359.0	17.5	16.8		360, 376, 341
E. coli		DMSO	-		62.0	9.2	-		64, 70, 52
		Test item	10		57.0	7.9	0.9		54, 51, 66
			33		53.7	7.0	0.9		61, 47, 53
			100		56.3	8.4	0.9		52, 66, 51
			333		66.3	6.0	1.1		72 P, 60 P, 67 P
			1000		70.0	5.3	1.1		68 P, 66 P, 76 P
			1500		54.0	5.3	0.9		56 P, 48 P, 58 P
		4-NQO	5		383.3	40.3	6.2		403, 337, 410

P = precipitation, C = contamination

Table 4:Preincubation assay, with metabolic activation

strain		test group	Dose (μg/plate)		Mean revertants per plate	SD	Factor	Individual revertant colony counts
TA 1535		DMSO Test item	- 10		- -	- -	- -	- C, - C, - C - C, - C, - C
			33		-	-	-	- C, - C, - C
			100		-	-	-	- C, - C, - C
			333		-	-	-	- C P, - C P, - C P
			1000		-	-	-	- C P, - C P, - C P
			1500		-	-	-	- C P, - C P, - C P
		2-AA	2.5		339.3	24.6	-	320, 331, 367
TA 100		DMSO	-		62.0	10.4	-	67, 69, 50
		Test item	10		53.7	7.5	0.9	46, 61, 54
			33		62.3	6.5	1.0	69, 56, 62
			100		49.7	3.5	0.8	53, 50, 46
			333		54.0	15.6	0.9	45 P, 45 P, 72 P
			1000		56.7	3.8	0.9	55 P, 54 P, 61 P
			1500		63.3	15.5	1.0	46 P, 76 P, 68 P
		2-AA	2.5		1887.7	54.4	30.4	1950, 1850, 1863
TA 1537		DMSO	-		12.0	3.6	-	15, 13, 8
		Test item	10		11.3	3.1	0.9	14, 8, 12
			33		11.3	2.1	0.9	13, 12, 9
			100		10.7	3.2	0.9	13, 12, 7
			333		11.0	1.0	0.9	10 P, 12 P, 11 P
			1000		8.0	1.0	0.7	8 P, 7 P, 9 P
			1500		9.3	1.5	0.8	8 P, 9 P, 11 P
		2-AA	2.5		119.3	13.4	9.9	104, 125, 129
TA 98		DMSO	-		37.7	6.7	-	30, 41, 42
		Test item	10		32.0	3.6	0.8	28, 35, 33
			33		31.3	2.1	0.8	33, 32, 29
			100		30.0	4.6	0.8	34, 31, 25
			333		25.3	8.5	0.7	34 P, 17 P, 25 P
			1000		30.0	3.5	0.8	32 P, 26 P, 32 P
			1500		24.3	4.5	0.6	29 P, 24 P, 20 P
		2-AA	2.5		1610.0	66.0	42.7	1577, 1567, 1686
E. coli		DMSO	-		64.7	10.5	-	54, 65, 75
		Test item	10		70.7	7.5	1.1	63, 71, 78
			33		61.7	10.0	1.0	52, 61, 72
			100		66.7	13.4	1.0	57, 61, 82
			333		65.3	4.0	1.0	63 P, 70 P, 63 P
			1000		59.7	3.8	0.9	64 P, 57 P, 58 P
			1500		65.0	11.1	1.0	53 P, 67 P, 75 P
		2-AA	60		325.3	17.6	5.0	344, 323, 309

P = precipitation, C=contamination

Table 5: Repeat experiment because of contamination, preincubation assay with and without metabolic activation

strain	test group	Dose (μg/plate)	Mean revertants per plate	SD	Factor	Individual revertant colony counts
TA 1535	DMSO	-	10.3	5.9	-	17, 6, 8
	Test item	10	8.3	4.9	0.8	14, 6, 5
		33	12.0	3.6	1.2	13, 15, 8
without S9		100	11.0	3.5	1.1	13, 13, 7
		333	9.3	3.1	0.9	6 P, 12 P, 10 P
		1000	8.7	2.1	0.8	8 P, 7 P, 11 P
		1500	6.7	2.5	0.6	4 P, 7 P, 9 P
	MNNG	5.0	2329.0	172.0	225.4	2259, 2203, 2525

TA 1535	DMSO	-	10.3	0.6	-	10, 11, 10
	Test item	10	12.3	3.5	1.2	16, 9, 12
		33	9.3	4.0	0.9	7, 14, 7
		100	9.3	4.2	0.9	8, 6, 14
with S9		333	13.0	2.6	1.3	10 P, 14 P, 15 P
		1000	8.7	3.8	0.8	7 P, 13 P, 6 P
		1500	7.3	0.6	0.7	8 P, 7 P, 7 P
	2-AA	2.5	240.3	19.9	23.3	232, 263, 226

P = precipitation

Conclusions:

Interpretation of results (migrated information):
negative

The substance is not mutagenic in bacteria.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

Justification for selection of genetic toxicity endpoint
Only study available.