Quaternary ammonium compounds, C12-14-alkyl(hydroxyethyl)dimethyl, chlorides

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:

2002-03-13 to 2002-03-28

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

[See concordance under 1.1. of the CSR]

IUPAC Name:

[See concordance under 1.1. of the CSR]

Details on test material:

- name of testsubstance used in report: Praepagen HY
- Physical state: yellowish, liquid
- Batch No.: ESDB 01 496
- active ingrdient /purity: Fattyalkyl dimethylhydroxyethylammonium chloride/ ca. 40% in water
- Expiration date of the lot/batch: 2003-07-19
- Storage condition of test material: room temperature (20°C)
- pH: 5.5 - 8.5 (20°C, 10g/L)

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 100

Details on mammalian cell type (if applicable):

- Properly maintained: yes
- Periodically checked for karyotype stability: yes

Additional strain / cell type characteristics:

not specified

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 98

Details on mammalian cell type (if applicable):

- Properly maintained: yes
- Periodically checked for karyotype stability: yes

Additional strain / cell type characteristics:

not specified

Species / strain 3

Species / strain / cell type:

S. typhimurium TA 97

Details on mammalian cell type (if applicable):

- Properly maintained: yes
- Periodically checked for karyotype stability: yes

Additional strain / cell type characteristics:

not specified

Species / strain 4

Species / strain / cell type:

S. typhimurium TA 102

Details on mammalian cell type (if applicable):

- Properly maintained: yes
- Periodically checked for karyotype stability: yes

Additional strain / cell type characteristics:

not specified

Species / strain 5

Species / strain / cell type:

S. typhimurium TA 1535

Details on mammalian cell type (if applicable):

- Properly maintained: yes
- Periodically checked for karyotype stability: yes

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

Postmitochondrial (S9) fraction from livers of male wistar rats, induced with Phenobarbital (i.p.) and ß-Naphtoflavon orally.

Test concentrations with justification for top dose:

1. Study:
all strains (±S9) : 0.005- 0.016 - 0.05 - 0.16 - 0.5 (mg/plate)
2. Study:
TA 97a, TA 100 (±S9), TA 98, TA 102, TA 1535 (-S9): 0.0016 - 0.005- 0.016 - 0.05 - 0.16 (mg/plate)
TA 98, TA 102, TA 1535 (+S9): 0.016 - 0.05 - 0.16 - 0.5 - 1.6 (mg/plate)
Negative controls tested with aqua bidest.

Vehicle / solvent:

aqua bidest

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

aqua bidest

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: ICR 191

Remarks:

positive control in TA 97a (-S9)

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

aqua bidest

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2- Aminoanthracen

Remarks:

positive control in TA 97a, TA 98, TA 100, TA 1535 (+S9)

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

aqua bidest

True negative controls:

no

Positive controls:

yes

Positive control substance:

cumene hydroperoxide

Remarks:

positive control in TA 102 (–S9)

Controls 4

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

aqua bidest

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylendiamine

Remarks:

positive control in TA 98 (– S9)

Controls 5

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

aqua bidest

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Nitrofurantoine

Remarks:

positive control in TA 100 (– S9)

Controls 6

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

aqua bidest

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Sodiumazide

Remarks:

positive control inTA 1535 (– S9)

Controls 7

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

aqua bidest

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Danthron

Remarks:

positive control in TA 102 (+S9)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation);
- Two independent studies were conducted
- Exposure duration: 48h
- NUMBER OF REPLICATIONS: 3 replicates per concentration level and control

Evaluation criteria:

Reduced background lawn is regarded to be a cytotoxic effect. Plates with reduced background lawn were not included into evaluation.
Mean values and standard deviations were calculated from colonies per plate of three replicates.
Induction rate = revertant colonies of test item divided by revertant colonies of the control.
The test item is to be interpretated mutagenic if there is a concentration effect relationship and the induction rate is ≥ 2.

Statistics:

The arithmetic mean value and the standard deviation were calculated out of three replicates of the revertant colony plates

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA 97, TA 98, TA 100, TA 1535, TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Based on the results of a preliminary test (non GLP) the test concentrations have been selected.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Mutagenic and Cytotoxic Effects of the Test Item Praepagen HY

Strain	S9	TestedConcentrationRange[mg/plate]	Lowest Mutagenic Concentration [mg/plate]	Lowest Cytotoxic Concentration [mg/plate]
TA 97a	-	0.0016 – 0.5	none	0.5
	+	0.0016 – 0.5	none	0.16
TA 98	-	0.0016 – 0.5	none	0.5
	+	0.005 – 1.6	none	1.6
TA 100	-	0.0016 – 0.5	none	0.5
	+	0.0016 – 0.5	none	0.5
TA 102	-	0.0016 – 0.5	none	0.16
	+	0.005 – 1.6	none	1.6
TA 1535	-	0.0016 – 0.5	none	0.5
	+	0.005 – 1.6	none	1.6

 

  Table 2: Summary of mean number of revertants (mean of 3 plates) in Salmonella typhimurium strains with and without metabolic activation

Experiment I
Concentration	TA1535		TA102		TA100		TA98		TA97a
[mg/plate]	- S9	+ S9	- S9	+ S9	- S9	+ S9	- S9	+ S9	- S9	+ S9
Control	24	12	322	382	123	133	22	24	234	275
0.5	n.c.	10	n.c.	353	n.c.	n.c.	n.c.	29.	n.c.	n.c.
0.16	22	13	333	450	74	101	40	40	211	238
0.05	30	15	325	480	126	113	45	35	278	276
0.016	29	15	397	457	131	100	42	35	277	259
0.005	29	13	344	461	133	100	32	34	263	233
Experiment II
	TA1535		TA102		TA100		TA98		TA97a
	- S9	+ S9	- S9	+ S9	- S9	+ S9	- S9	+ S9	- S9	+ S9
Control	25	13	389	501	117	112	25	30	260	277
1.6	—	n.c.	—	n.c.	—	—	—	—	—	—
0.5	—	10	—	203	—	—	—	—	—	—
0.16	17	10	198	346	57	129	28	n.c.	100	n.c.
0.05	12	11	315	333	53	102	31	15	219	121
0.016	25	14	273	364	122	110	34	33	232	226
0.005	16	—	218	—	72	100	21	35	231	205
0.0016	30	—	339	—	115	93	28	32	116	136

n.c.= not counted, due to reduced background lawn; — = concentration not tested

Applicant's summary and conclusion

Conclusions:

Based on the results of this study the test item was found to have no mutagenic effects on Salmonella typhimurium strains TA 97a, TA 100, TA 98, TA 102, TA 1535 with (+) and without (-) the metabolic activation system S9 from Wistar rats at concentrations up to 0.5mg/plate.