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Toxicological information

Genetic toxicity: in vitro

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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 guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
1983
Deviations:
no
Qualifier:
according to
Guideline:
other: EU Annex V Method 431
Deviations:
not specified
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
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 / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
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 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: bidest water
- Justification for choice of solvent/vehicle: none given in test report
Controlsopen allclose all
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
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
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
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
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
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 resultsopen allclose all
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
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
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
without
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:
(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.