(3-chloro-2-hydroxypropyl)trimethylammonium chloride

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:

1984-01-12 to 1984-04-02

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to an appropriate OECD test guideline, with acceptable restrictions. The restrictions were: the range of strains does not comply with the current guideline.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1984

Report date:

1984

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1983

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EU Annex V Method 431

Deviations:

not specified

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

(3-chloro-2-hydroxypropyl)trimethylammonium chloride

EC Number:

222-048-3

EC Name:

(3-chloro-2-hydroxypropyl)trimethylammonium chloride

Cas Number:

3327-22-8

Molecular formula:

C6H15ClNO.Cl

IUPAC Name:

3-chloro-2-hydroxy-N,N,N-trimethylpropan-1-aminium chloride

Details on test material:

- Name of test material (as cited in study report): 3-chlor-2-hydroxypropyltrimethyl-ammonium chloride
- Physical state: solid
- Analytical purity: dry: 94.59%, humid: 65.51%
- Lot/batch No.: 0201
- Stability under test conditions: stable
- Storage condition of test material: at room temperature in the dark

Method

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:

Aroclor induced rat liver S9

Test concentrations with justification for top dose:

ranging from 1.58 - 5000 ug/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: bidest water
- Justification for choice of solvent/vehicle: none given in test report

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

TA 100 without activation

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA 1535 without activation

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

TA 98 TA 100 TA 1537 TA 1538 without activation

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: N-nitrosoguanidine

Remarks:

TA 98 100 1535 without activation

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

all strains with activation

Controls 6

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

TA 98 TA100 TA 1537 TA 1538 with activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation);

DURATION
- Exposure duration: 3 days

NUMBER OF REPLICATIONS: triplicate

DETERMINATION OF CYTOTOXICITY
- Method: other: relative survival rate: comparison of number of revertants in presence and absence of prototrophic bacterai (spontaneous revertants of TA 1573 (RTA))

OTHER: To avoid any light effects on the test compound, all experimentation was performed under yellow light.

Evaluation criteria:

A positive result is a reproducible dose effect with a 2-fold increase in the number of revertants over controls in at least one strain. A 1.5-fold increase is the criterion for a positive result in strain TA 100.

Statistics:

The number of revertants was counted with a Fischer 880 counter unless precipitation had occurred. Mean number of revertants per plate was calculated.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

dose dependent

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 98

Metabolic activation:

without

Genotoxicity:

positive

Remarks:

at highest dose

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 1537

Genotoxicity:

positive

Remarks:

at two highest doses tested

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: none reported
- The pH value is not reported
- Other confounding effects: none reported

COMPARISON WITH HISTORICAL CONTROL DATA: none given in study report

ADDITIONAL INFORMATION ON CYTOTOXICITY: neither quantitative nor qualitative evidence of cytotoxic effects was observed

Any other information on results incl. tables

Table 1: Dose range-finding study Relative survival rate

-	Without activation			With activation
Conc. (µg/plate)	TA1537	TA1537 + RTA	Relative survival rate	TA1537	TA1537 + RTA	Relative survival rate
0*	6	190	1.00	16	226	1.00
1.58	8	214	1.12	16	236	1.05
5	7	197	1.03	14	219	0.90
15.8	10	205	1.06	14	206	0.91
50	10	202	1.04	15	235	1.05
158	8	197	1.03	13	224	1.00
500	8	204	1.07	17	237	1.05
1580	13	233	1.20	13	232	1.04
5000	17	214	1.07	17	232	1.02

*solvent control with water

RTA: spontaneous revertants of TA 1573

 

Table 2: Experiment 1 Plate incorporation Number of revertants per plate (mean of 3 plates)

-	TA98			TA100			TA1535
Conc. (µg/plate)	- MA	+ MA	Cytotoxic (yes/no)	- MA	+ MA	Cytotoxic (yes/no)	- MA	+ MA	Cytotoxic (yes/no)
0*	18	29	no	185	153	no	15	10	no
1.58	23	30	no	182	116	no	20	9	no
5	23	30	no	185	138	no	17	12	no
15.8	22	28	no	198	145	no	19	9	no
50	23	30	no	204	147	no	13	15	no
158	26	30	no	185	141	no	21	14	no
500	34	31	no	162	183	no	28	18	no
1580	30	30	no	178	158	no	40	21	no
5000	39	28	no	175	168	no	68	41	no
Positive control 1	462	75	no	305	256	no	-	82	no
Positive control 2	166	208	no	281	532	no	227	-	no

*solvent control with water

 

Table 3 Experiment 1 Plate incorporation Number of revertants per plate (mean of 3 plates)

-	TA1537			TA1538
Conc. (µg/plate)	- MA	+ MA	Cytotoxic (yes/no)	- MA	+ MA	Cytotoxic (yes/no)
0*	6	16	no	14	21	no
1.58	8	16	no	18	20	no
5	7	14	no	12	16	no
15.8	10	14	no	23	18	no
50	10	15	no	21	21	no
158	8	13	no	12	17	no
500	8	17	no	16	19	no
1580	13	13	no	15	18	no
5000	17	17	no	14	18	no
Positive control 1	569	168	no	-	76	no
Positive control 2	122	106	no	835	103	no

*solvent control with water

 

Table 4: Experiment 2 Repeat of strain TA 1537 Number of revertants per plate (mean of 3 plates)

-	TA100
Conc. (µg/plate)	- MA	Cytotoxic (yes/no)
0*	20	no
1.58	19	no
5	24	no
15.8	26	no
50	28	no
158	25	no
500	36	no
1580	46	no
5000	80	no
Positive control	781	no

*solvent control with water

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
positive with and without activation

(3-Chloro-2-hydroxypropyl)trimethylammonium chloride was tested in OECD 471 under GLP. A dose-dependant increase in the number of revertants was observed in Salmonella typhimurium TA1535 both with and without activation. An increase in the number of revertants was also seen in strain TA 98 in the absence of activation at the highest dose tested, and in TA 1537 without activation at the two highest doses tested. It was concluded that the test substance is mutagenic under the conditions of the test.