trisodium ({3-[(carboxylatomethyl)amino]propyl}{3-[(carboxylatomethyl)(C16-18, C18 unsat alkyl)amino]propyl}amino)acetate

Genetic toxicity: in vitro

Administrative data

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:

8 January 1993 - 26 January 1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study was performed under GLP and according to international guidelines. However in the current guideline E.coli WP2 strains or S. typhimurium TA102 have to be included. With this test it is not possible to identify certain oxidising mutagens, cross-linking agents and hydrazines. Such substances may be detected by E.coli WP2 strains or S. typhimurium TA102 which have an AT base pair at the primary reversion site in stead of GC base pairs which the strains tested in this study have.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

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

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OTS 798.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)

Principles of method if other than guideline:

In the current guideline E.coli WP2 strains or S. typhimurium TA102 have to be included. With this test it is not possible to identify certain oxidising mutagens, cross-linking agents and hydrazines. Such substances may be detected by E.coli WP2 strains or S. typhimurium TA102 which have an AT base pair at the primary reversion site in stead of GC base pairs which the strains tested in this study have.

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Amines, N-C8-22-alkylpolytrimethylenepoly-, carboxymethyl derivs., sodium salts

EC Number:

307-458-3

EC Name:

Amines, N-C8-22-alkylpolytrimethylenepoly-, carboxymethyl derivs., sodium salts

Cas Number:

97659-53-5

Details on test material:

Name: Ampholak 7TX
Chemicals name: Amines, N-tallowalkyl-polytrimethylene-poly-, carboxymethylderivates, sodium salt
CAS No.: 97659-53-5
Batch No.: BX 3719
Appearance: pale yellow viscous liquid
Storage: clear glass jar, ambient room temperature in the dark

Method

Target gene:

Gene coding for histidine

Species / strain

Species / strain / cell type:

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

Additional strain / cell type characteristics:

other: see below in section any other information on materials and methods

Metabolic activation:

with and without

Metabolic activation system:

S9-mix

Test concentrations with justification for top dose:

50, 158, 500, 1580 and 5000 µg/plate

Vehicle / solvent:

puirifed water

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: sodium azide, 2-aminoanthracene, 9-aminoacridine, 2-nitrofuorene, Benzo[a]pyrene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 2 days

SELECTION AGENT (mutation assays): histidine defiecient medium

NUMBER OF REPLICATIONS: Triplicate plates per concentration

NUMBER OF CELLS EVALUATED: After incubation, numbers of revertant colonies were counted manually. Total colonies on nutrient plates were counted in the same way. Growth of the background lawn of non-revertant cells on minimal plates was verified.

DETERMINATION OF CYTOTOXICITY
- Method: a study was per formed with strain TA98 at 2.5, 5, 25, 50, 250, 500, 2500 and 5000 µg/plate, otherwise same methods as the main study.

Evaluation criteria:

no data

Statistics:

no data

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

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Mutation test 1

strain	Dose level (µg/plate)	Metabolic activation	Mean revertant colony counts	SD
TA98	5000	+	36	3
	1580	+	42	12
	5500	+	42	4
	158	+	39	5
	50	+	37	5
	0	+	37	6
	5000	-	25	5
	1580	-	22	2
	5500	-	26	3
	158	-	28	6
	50	-	25	1
	0	-	24	4
TA100	5000	+	121	3
	1580	+	130	12
	5500	+	122	9
	158	+	120	20
	50	+	113	4
	0	+	126	12
	5000	-	69	20
	1580	-	139	10
	5500	-	107	4
	158	-	108	6
	50	-	110	6
	0	-	138	10
TA1535	5000	+	12	3
	1580	+	11	3
	5500	+	16	2
	158	+	13	3
	50	+	9	1
	0	+	12	1
	5000	-	0	0
	1580	-	9	2
	5500	-	8	2
	158	-	15	2
	50	-	12	3
	0	-	11	3
TA1537	5000	+	15	3
	1580	+	12	3
	5500	+	9	4
	158	+	6	2
	50	+	8	3
	0	+	12	3
	5000	-	13	2
	1580	-	11	3
	5500	-	9	1
	158	-	6	3
	50	-	8	4
	0	-	8	2

- Absence

+ Presence

SD Standard deviation

Positive control Mutation test 1:

Strain	Compound	Dose level (µg/plate)	Metabolic activation	Mean revertant colony counts	SD
TA98	Benzo[a]pyrene	5	-	22	9
TA98	Benzo[a]pyrene	5	+	544	29
TA98	2-nitrofluorne	1	-	138	5
TA100	Benzo[a]pyrene	5	-	110	10
TA100	Benzo[a]pyrene	5	+	829	70
TA100	Sodium azide	2	-	969	14
TA1535	2-aminoanthracene	2	-	153	47
TA1535	2-aminoanthracene	2	+	247	44
TA1535	Sodium azide	2	-	180	11
TA1537	Benzo[a]pyrene	5	-	14	3
TA1537	Benzo[a]pyrene	5	+	222	23
TA1537	9-aminoacridine	80	-	201	75

- Absence

+ Presence

SD Standard deviation

Mutation test 2:

strain	Dose level (µg/plate)	Metabolic activation	Mean revertant colony counts	SD
TA98	5000	+	35	9
	1580	+	41	3
	5500	+	41	8
	158	+	45	11
	50	+	36	5
	0	+	40	7
	5000	-	15	3
	1580	-	21	5
	5500	-	22	4
	158	-	25	5
	50	-	22	6
	0	-	30	9
TA100	5000	+	76	15
	1580	+	113	6
	5500	+	105	3
	158	+	80	12
	50	+	74	4
	0	+	98	7
	5000	-	15	3
	1580	-	73	16
	5500	-	93	5
	158	-	90	11
	50	-	89	3
	0	-	91	6
TA1535	5000	+	11	4
	1580	+	7	2
	5500	+	8	1
	158	+	9	3
	50	+	13	4
	0	+	11	4
	5000	-	2	1
	1580	-	6	2
	5500	-	11	1
	158	-	9	3
	50	-	9	1
	0	-	10	3
TA1537	5000	+	2	1
	1580	+	9	4
	5500	+	11	2
	158	+	8	1
	50	+	9	4
	0	+	11	2
	5000	-	7	1
	1580	-	3	1
	5500	-	9	3
	158	-	8	3
	50	-	7	1
	0	-	7	1

- Absence

+ Presence

SD Standard deviation

                                                                                 

Positive control Mutation test 2:

Strain	Compound	Dose level (µg/plate)	Metabolic activation	Mean revertant colony counts	SD
TA98	Benzo[a]pyrene	5	-	22	5
TA98	Benzo[a]pyrene	5	+	525	46
TA98	2-nitrofluorne	1	-	147	28
TA100	Benzo[a]pyrene	5	-	79	17
TA100	Benzo[a]pyrene	5	+	565	219
TA100	Sodium azide	2	-	723	151
TA1535	2-aminoanthracene	2	-	9	0
TA1535	2-aminoanthracene	2	+	255	94
TA1535	Sodium azide	2	-	938	14
TA1537	Benzo[a]pyrene	5	-	6	3
TA1537	Benzo[a]pyrene	5	+	189	21
TA1537	9-aminoacridine	80	-	169	70

- Absence

+ Presence

SD Standard deviation

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Ampholak 7TX was devoid of mutagenic activity under the conditions of test.

Executive summary:

An Ames test was performed according to OECD471, EPA Toxic Substances Control Act Test Guideline 798.5265 (1987) and EC Annex V B.14. The study complies with GLP. The studies, which were conducted in the presence and absence of an activating system derived from rat liver (S-9 mix), employed a range of levels of Ampholak 7TX in purified water from 50 to 5000 µg per plate, selected following a preliminary toxicity test in strain TA98. All tests included solvent (purified water) controls with and without S-9 mix.No increases in reversion to prototrophy were obtained with any of the four bacterial strains at the Ampholak 7TX levels tested, either in the presence or absence of S-9 mix. Inhibition of growth, observed as a reduction in revertant colony numbers, occurred in all strains following exposure to Ampholak 7TX at 5000 µg per plate on at least one occasion of testing.

Marked increases in the number of revertant colonies were induced by the known mutagens benzo[a]pyrene, 2 -nitrofluorene, 2 -aminoanthracene, 9 -aminoacridine, and sodium azide when examined under similar conditions. It was concluded that Ampholak 7TX was devoid of mutagenic activity under the conditions of test.