Disodium [[N,N'-ethylenebis[N-(carboxymethyl)glycinato]](4-)-N,N',O,O',ON,ON']cuprate(2-)

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:

1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Well performed and reported GLP study; however only using strains Salmonella strains TA 98, 100, 1535 and 1537 and not the E coli strain (not required in 1992).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Disodium [[N,N'-ethylenebis[N-(carboxymethyl)glycinato]](4-)-N,N',O,O',ON,ON']cuprate(2-)

EC Number:

237-864-5

EC Name:

Disodium [[N,N'-ethylenebis[N-(carboxymethyl)glycinato]](4-)-N,N',O,O',ON,ON']cuprate(2-)

Cas Number:

14025-15-1

Molecular formula:

C10H12CuN2O8.2Na

IUPAC Name:

Copper(2+) ion disodium 2-({2-[bis(carboxylatomethyl)amino]ethyl} (carboxylatomethyl)amino)acetate

Test material form:

other: solution

Details on test material:

Name: Trilon B-CU 1550
Batch no.: Labor Journal No. 91 233
Test substance no.: 91/401
Purity: 47.2% (52.1% water)
Appearance: blue liquid

Method

Target gene:

Point mutation

Species / strain

Species / strain / cell type:

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

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced rat liver S9 mix

Test concentrations with justification for top dose:

First (standard plate test) and second (preincubation) test: 0, 100, 500, 2500, 5000 and 10000 ug/plate

Vehicle / solvent:

water

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

other: 2-aminoanthracene, N-methyl-N-nitro-N-nitrososguanidine, nitro-o-phenylendiamine

Remarks:

2-AA was used with S9-mix, the other without S9-mix

Details on test system and experimental conditions:

METHOD OF APPLICATION: standard plate test (Ames) and preincubation test (Yagahi et al. and Matsushima et al.)

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3 test plates per dose

NUMBER OF CELLS EVALUATED: colonies are counted

DETERMINATION OF CYTOTOXICITY
- Method: his- background growth

Evaluation criteria:

Positive result in case of:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results

Statistics:

No info

Results and discussion

Test results

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

See for test results the Tables below

Remarks on result:

other: strain/cell type:

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Results of Strain TA 1535 (standard plate)

 

Dose (ug/plate)	Number of revertants per plate
	-S9-mix		+S9-mix
	Mean	SD	Mean	SD
0	19	3	22	3
100	17	5	22	2
500	18	3	22	2
2500	22	4	24	3
5000	17	2	22	1
10000	17	3	13	0
2-AA	-	-	145	7
MNNG	1017	6	-	-

 

Results of Strain TA 100 (standard plate)

 

Dose (ug/plate)	Number of revertants per plate
	-S9-mix		+S9-mix
	Mean	SD	Mean	SD
0	124	11	144	6
100	124	21	149	4
500	134	19	214	15
2500	124	21	152	16
5000	131	6	133	10
10000	126	17	146	4
2-AA	-	-	1173	170
MNNG	1037	96	-	-

 

Results of Strain TA 1537 (standard plate)

 

Dose (ug/plate)	Number of revertants per plate
	-S9-mix		+S9-mix
	Mean	SD	Mean	SD
0	8	2	10	3
100	8	3	12	2
500	8	2	14	2
2500	8	3	10	1
5000	9	1	9	4
10000	7	4	9	1
2-AA	-	-	116	6
AAC	563	46	-	-

 

Results of Strain TA 98 (standard plate)

 

Dose (ug/plate)	Number of revertants per plate
	-S9-mix		+S9-mix
	Mean	SD	Mean	SD
0	18	3	32	4
100	16	6	33	7
500	22	4	32	2
2500	22	1	26	6
5000	23	2	18	3
10000	24	1	23	7
2-AA	-	-	738	58
NPD	688	32	-	-

 

Results of Strain TA 1535 (preincubation)

 

Dose (ug/plate)	Number of revertants per plate
	-S9-mix		+S9-mix
	Mean	SD	Mean	SD
0	16	4	13	3
100	14	2	11	1
500	15	3	9	1
2500	12	3	11	3
5000	14	5	11	3
10000	11	5	11	2
2-AA	-	-	82	3
MNNG	571	30	-	-

 

Results of Strain TA 100 (preincubation)

 

Dose (ug/plate)	Number of revertants per plate
	-S9-mix		+S9-mix
	Mean	SD	Mean	SD
0	100	14	107	5
100	95	12	106	3
500	98	15	124	5
2500	123	5	107	8
5000	107	14	126	3
10000	109	12	128	19
2-AA	-	-	561	9
MNNG	578	66	-	-

 

Results of Strain TA 1537 (preincubation)

 

Dose (ug/plate)	Number of revertants per plate
	-S9-mix		+S9-mix
	Mean	SD	Mean	SD
0	8	1	12	2
100	8	3	9	1
500	7	3	9	2
2500	6	2	7	2
5000	7	0	6	2
10000	5	3	8	0
2-AA	-	-	78	9
AAC	412	14	-	-

 

Results of Strain TA 98 (preincubation)

 

Dose (ug/plate)	Number of revertants per plate
	-S9-mix		+S9-mix
	Mean	SD	Mean	SD
0	28	5	27	7
100	22	3	29	2
500	22	2	26	9
2500	17	1	25	2
5000	26	3	24	1
10000	25	2	23	2
2-AA	-	-	556	39
NPD	744	38	-	-

 

 

Applicant's summary and conclusion

Conclusions:

The test substance EDTA-CuNa2 is not mutagenic in the Ames test under the experiemnatl conditions used.

Executive summary:

The test substance EDTA-CuNa2 (Trilon B-CU 1550) was tested for its mutagenic potential based on the ability to induce back mutations in selected loci in several strains of Salmonelle thyphimurium (TA 1535, TA 100, TA 1537 and TA 98) in the Ames test. The dose range was between 100 and 10000 ug/plate. The standard plate test and the preincubation test were used both with and without metabolic activation (Aroclor induced rat liver S9 -mix). No precipitation of the test substance was found. No bacteriotoxic effect was observed. An increase in the number of His+ revertants was not observed both in the standard plate test and in the pre-incubation test either with or without S9 -mix. Based on the results of this study, the test substance EDTA-CuNa2 is not mutagenic in the Ames test under the experiemnatl conditions used.