1-Propanaminium, N-(3-aminopropyl)-2-hydroxy-N,N-dimethyl-3-sulfo-, N-(C8-18(even numbered) acyl) derivs., hydroxides, inner salts

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:

August 1996 - January 1997

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

Bacterial strains tested not in accordance with the current recommendations (TA1538 instead of TA102), less than five different analysable concentrations of the test substance used

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

(2-hydroxy-3-sulphopropyl)dimethyl[3-[(1-oxooctyl)amino]propyl]ammonium hydroxide

Molecular formula:

C16H34N2O5S

IUPAC Name:

(2-hydroxy-3-sulphopropyl)dimethyl[3-[(1-oxooctyl)amino]propyl]ammonium hydroxide

Constituent 2

Reference substance name:

(2-hydroxy-3-sulphopropyl)dimethyl[3-[(1-oxodecyl)amino]propyl]ammonium hydroxide

Molecular formula:

C18H38N2O5S

IUPAC Name:

(2-hydroxy-3-sulphopropyl)dimethyl[3-[(1-oxodecyl)amino]propyl]ammonium hydroxide

Constituent 3

Reference substance name:

(2-hydroxy-3-sulphopropyl)dimethyl[3-[(1-oxododecyl)amino]propyl]ammonium hydroxide

EC Number:

242-893-1

EC Name:

(2-hydroxy-3-sulphopropyl)dimethyl[3-[(1-oxododecyl)amino]propyl]ammonium hydroxide

Cas Number:

19223-55-3

Molecular formula:

C20H42N2O5S

IUPAC Name:

N-[3-(dodecanoylamino)propyl]-2-hydroxy-N,N-dimethyl-3-sulfopropan-1-aminium hydroxide

Constituent 4

Reference substance name:

(2-hydroxy-3-sulphopropyl)dimethyl[3-[(1-oxotetradecyl)amino]propyl]ammonium hydroxide

Molecular formula:

C22H46N2O5S

IUPAC Name:

(2-hydroxy-3-sulphopropyl)dimethyl[3-[(1-oxotetradecyl)amino]propyl]ammonium hydroxide

Constituent 5

Reference substance name:

(2-hydroxy-3-sulphopropyl)dimethyl[3-[(1-oxohexadecyl)amino]propyl]ammonium hydroxide

Molecular formula:

C24H50N2O5S

IUPAC Name:

(2-hydroxy-3-sulphopropyl)dimethyl[3-[(1-oxohexadecyl)amino]propyl]ammonium hydroxide

Constituent 6

Reference substance name:

[2-hydroxy-3-sulphopropyl]dimethyl[3-[(1-oxooctadecyl)amino]propyl]ammonium hydroxide

EC Number:

264-390-6

EC Name:

[2-hydroxy-3-sulphopropyl]dimethyl[3-[(1-oxooctadecyl)amino]propyl]ammonium hydroxide

Cas Number:

63663-12-7

Molecular formula:

C26H54N2O5S

IUPAC Name:

2-hydroxy-N,N-dimethyl-N-[3-(stearoylamino)propyl]-3-sulfopropan-1-aminium hydroxide

Constituent 7

Reference substance name:

Water

EC Number:

231-791-2

EC Name:

Water

Cas Number:

7732-18-5

Molecular formula:

H2O

IUPAC Name:

Water

impurity 1

Reference substance name:

Sodium chloride

EC Number:

231-598-3

EC Name:

Sodium chloride

Cas Number:

7647-14-5

Molecular formula:

ClNa

IUPAC Name:

sodium chloride

impurity 2

Reference substance name:

Glycerol

EC Number:

200-289-5

EC Name:

Glycerol

Cas Number:

56-81-5

Molecular formula:

C3H8O3

IUPAC Name:

glycerol

impurity 3

Reference substance name:

Disodium 2-hydroxypropane-1,3-disulfonate

Molecular formula:

C3H8O7S2.Na2

IUPAC Name:

Disodium 2-hydroxypropane-1,3-disulfonate

impurity 4

Reference substance name:

Sodium (±)-2,3-dihydroxypropanesulphonate

EC Number:

252-542-4

EC Name:

Sodium (±)-2,3-dihydroxypropanesulphonate

Cas Number:

35396-47-5

Molecular formula:

C3H8O5S.Na

IUPAC Name:

sodium (±)-2,3-dihydroxypropanesulphonate

impurity 5

Reference substance name:

Amides, C8-18 even numbered, N-[3-(dimethylamino)propyl]

EC Number:

930-947-3

IUPAC Name:

Amides, C8-18 even numbered, N-[3-(dimethylamino)propyl]

Test material form:

liquid

Method

Target gene:

His gene

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

S9 mix from Aroclor 1254-induced rat liver

Test concentrations with justification for top dose:

0.002, 0.002, 0.2, 2 and 20 µL test solution/plate (1st experiment)
0.02, 0.06, 0.2, 0.6 and 2 µL test solution/plate (2nd experiment)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: 50% aqueous solution of test substance used

Controls

Untreated negative controls:

yes

Remarks:

water

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

other: 2-aminoanthracene, 2-aminofluorene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: not applicable
- Exposure duration: incubation at 37°C for 2 days

SELECTION AGENT (mutation assays): 0.05 mM L-histidine

NUMBER OF REPLICATIONS: 3

Evaluation criteria:

A test material is considered mutagenic if there is a reproducibly increasing dose-response curve of induced revertant colonies for at least 3 test concentrations. The minimal criteria for a positive response are a 2- to 3-fold increase in the number of revertants (at least 15 colonies) over the spontaneous number for the TA1535, TA1537, TA1538 and TA98 strains, and a 50% increase for the TA100 strain. In addition, a positive response must not be observed only at concentrations near toxic dose levels.

Statistics:

Mean and standard deviation calculation

Results and discussion

Test results

Test results 1

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:

cytotoxicity

Remarks:

from and above 0.6 µL/plate in the absence of S9, from and above 2 µL/plate (0.6 µL/plate for TA100) in the presence of S9

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

from and above 0.6 µL/plate in the absence of S9, from and above 2 µL/plate in the presence of S9

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

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

Concentration (µL test solution)	TA98						TA100						TA1535						TA1537						TA1538
	-S9			+S9			-S9			+S9			-S9			+S9			-S9			+S9			-S9			+S9
	Mean	SD	Ratio vs. controls	Mean	SD	Ratio vs. controls	Mean	SD	Ratio vs. controls	Mean	SD	Ratio vs. controls	Mean	SD	Ratio vs. controls	Mean	SD	Ratio vs. controls	Mean	SD	Ratio vs. controls	Mean	SD	Ratio vs. controls	Mean	SD	Ratio vs. controls	Mean	SD	Ratio vs. controls
0 (water)	18	1	1.0	27	7	1.0	169	10	1.0	175	13	1.0	11	4	1.0	13	1	1.0	13	2	1.0	12	5	1.0	9	2	1.0	14	2	1.0
0.002	19	3	1.0	28	9	1.1	152	14	0.9	165	29	0.9	12	2	1.2	11	2	0.9	14	5	1.1	12	4	0.9	12	3	1.3	19	3	1.4
0.02	22	2	1.2	23	6	0.9	173	8	1.0	162	5	0.9	14	1	1.3	10	3	0.8	16	6	1.2	16	2	1.3	8	3	0.9	17	3	1.2
0.2	19	6	1.1	27	2	1.0	148	33	0.9	196	24	1.1	14	3	1.3	16	1	1.3	12	4	0.9	18	6	1.4	15	2	1.6	22	7	1.6
2	0*	0	0.0	0*	0	0.0	0*	0	0.0	0*	0	0.0	0*	0	0.0	0*	0	0.0	0*	0	0.0	0*	0	0.0	0*	0	0.0	0*	0	0.0
20	0*	0	0.0	0*	0	0.0	0*	0	0.0	0*	0	0.0	0*	0	0.0	0*	0	0.0	0*	0	0.0	0*	0	0.0	0*	0	0.0	0*	0	0.0
Positive controls
2-Nitrofluorene	565	21	30.8	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	1600	50	177.8	-	-	-
2-Aminofluorene	-	-	-	318	18	11.9	-	-	-	1175	65	6.7	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Sodium azide	-	-	-	-	-	-	526	27	3.1	-	-	-	439	12	41.2	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
2-Aminoanthracene	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	177	7	13.6	-	-	-	145	10	11.7	-	-	-	1375	56	98.2
9-Aminoacridine	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	263	14	19.7	-	-	-	-	-	-	-	-	-

* Cytotoxicity

First experiment: Summary of numbers of revertants per plate

Applicant's summary and conclusion

Conclusions:

Cocamidopropyl hydroxysultaine, as a 50% aqueous solution, was not mutagenic in a bacterial reverse mutation assay up to cytotoxic concentrations, in the presence or in the absence of metabolic activation.

Executive summary:

Cocamidopropyl hydroxysultaine, as a 50% aqueous solution, was tested in a bacterial reverse mutation (Ames) test using the Salmonella typhimurium TA1535, TA1537, TA1538, TA98 and TA100 strains. The bacterial strains were exposed on minimal agar plates (using a plate incorporation method) to a range of concentrations up to 20 µL of test solution per plate, both in the presence or absence of an exogenous metabolic activation system, consisting of S9 mix from Aroclor 1254 -induced rat liver. Two independent experiments were performed in triplicate.

In this Ames test, no significant increase in the mean number of revertants over the respective vehicle controls was observed in any of the bacterial strains tested, either in the presence or in the absence of metabolic activation, up to 0.2 or 0.6 µL test solution/plate depending on the strain. Cyotoxic effects were observed at higher dose levels.

Under the conditions of this assay, Cocamidopropyl hydroxysultaine, as a 50% aqueous solution, was not mutagenic up to cytotoxic concentrations.