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REACH

3,5,8,10,13,15,18,20,23,25,28,30,33,35,38,40,41,43,45,47,51,53,55,57,59,61,63, 65,67,69,71,73-dotriacontazapentacosacyclo [35.3.3.36,7.311, 12.316,17.321,22.326,27.331,32.22,41.236,43.13,40.15,8.110,13.115,18.120,23.125,28.130,33.135,38.145,51.147,73.153,55.157,59.161,63.165,67.169,71] octacontane-44,46,48,49,50,52,54,56,58,60,62,64,66,68,70,72-hexadecone, hydrochloride hydrate

EC number: 700-970-1 | CAS number: -

Life Cycle description
Uses advised against

Toxicological information

Genetic toxicity: in vitro

Administrative data

Endpoint:: in vitro gene mutation study in bacteria
Type of information:: experimental study
Adequacy of study:: key study
Reliability:: 1 (reliable without restriction)
Rationale for reliability incl. deficiencies:: guideline study

Data source

Reference

Reference Type:: study report
Title:: Unnamed
Year:: 2015
Report date:: 2015

Materials and methods

Test guidelineopen all close all

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:: EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
Deviations:: no

Test guideline 3

Qualifier:: according to guideline
Guideline:: EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:: no

GLP compliance:: yes (incl. QA statement)
Remarks:: OGYI/38593-5/2012
Type of assay:: bacterial reverse mutation assay

Test material

Method

Species / strainopen all close all

Species / strain 1

Species / strain / cell type:: S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:: not applicable

Species / strain 2

Species / strain / cell type:: E. coli WP2 uvr A
Additional strain / cell type characteristics:: not applicable

Metabolic activation:: with and without
Metabolic activation system:: cofactor-supplemented post-mitochondrial S9 fraction
Test concentrations with justification for top dose:: Based on the results of the preliminary experiment, the examined test concentrations in the main tests were 2500, 791, 250, 79.1, 25, 7.91, 2.5 and 0.791 μg/plate with and without metabolic activation. At the top dose precipitation and cytotoxicity were observed in all strains.

Controls

Untreated negative controls:: no
Negative solvent / vehicle controls:: yes
Remarks:: Dimethyl sulfoxide, water
True negative controls:: no
Positive controls:: yes
Positive control substance:: 9-aminoacridine; sodium azide; methylmethanesulfonate; other: 4-nitro-1,2-phenylenediamine; 2-aminoanthracene

Results and discussion

Test resultsopen all close all

Test results 1

: Key result
Species / strain:: S. typhimurium TA 1535
Metabolic activation:: with and without
Genotoxicity:: negative
Cytotoxicity / choice of top concentrations:: cytotoxicity
Vehicle controls validity:: valid
Untreated negative controls validity:: not applicable
True negative controls validity:: valid
Positive controls validity:: valid

Test results 2

: Key result
Species / strain:: S. typhimurium TA 1537
Metabolic activation:: with and without
Genotoxicity:: negative
Cytotoxicity / choice of top concentrations:: cytotoxicity
Vehicle controls validity:: valid
Untreated negative controls validity:: not applicable
True negative controls validity:: valid
Positive controls validity:: valid

Test results 3

: Key result
Species / strain:: S. typhimurium TA 98
Metabolic activation:: with and without
Genotoxicity:: negative
Cytotoxicity / choice of top concentrations:: cytotoxicity
Vehicle controls validity:: valid
Untreated negative controls validity:: not applicable
True negative controls validity:: valid
Positive controls validity:: valid

Test results 4

: Key result
Species / strain:: S. typhimurium TA 100
Metabolic activation:: with and without
Genotoxicity:: negative
Cytotoxicity / choice of top concentrations:: cytotoxicity
Vehicle controls validity:: valid
Untreated negative controls validity:: not applicable
True negative controls validity:: valid
Positive controls validity:: valid

Test results 5

: Key result
Species / strain:: E. coli WP2 uvr A
Metabolic activation:: with and without
Genotoxicity:: negative
Cytotoxicity / choice of top concentrations:: cytotoxicity
Vehicle controls validity:: valid
Untreated negative controls validity:: not applicable
True negative controls validity:: valid
Positive controls validity:: valid

Any other information on results incl. tables

Confirmatory Mutation Test (Pre-Incubation Method)

Test Item: CUCURBIT[8]URIL

Strain: Salmonella typhimurium TA98

Cell count (overnight culture): 2.01 x 10⁹CFU/mL

Concentration: (μg/plate)

Revertant colony number

Concentration: (μg/plate)

Revertant colony number

-S9

+S9

-S9

+S9

25000

2 SR, SP

6 SR, SP

4 R, SP

8 R, SP

Untreated control

Mean

3.3

2.31

0.19

5.3

2.31

0.19

Mean

22.7

4.73

1.28

31.7

3.06

1.10

791

12 SR

9 SR

6 SR

10 R

DMSO control

Mean

9.0

3.00

0.51

10.0

0.00

0.35

Mean

17.7

2.08

1.00

28.7

1.53

1.00

250

11 SR

13 SR

22SR

Positive control

NPD (4 μg)

293

312

302

2AA

(2 μg)

2366

2380

2374

Mean

15.3

5.86

0.87

27.7

3.06

0.97

Mean

302.3

9.50

17.11

2373.3

7.02

82.79

79.1

SP: Slight precipitate

R: Reduced background lawn

SR: Slightly reduced background lawn

MF: Mutation factor

SD: Standard deviation

+S9: with S9 mix

-S9: without S9 mix

Mean revertants (test item)

Mutation factor = -------------------------------------

Mean revertants (solvent control)

Mean

20.0

3.46

1.13

38.7

1.53

1.35

Mean

24.0

5.57

1.36

33.0

1.00

1.15

7.91

Mean

26.3

4.04

1.49

31.3

5.51

1.09

2.5

Mean

21.7

7.23

1.23

30.0

6.56

1.05

0.791

Mean

19.0

2.00

1.08

22.7

4.73

0.79

Confirmatory Mutation Test (Pre-Incubation Method)

Test Item: CUCURBIT[8]URIL

Strain: Salmonella typhimurium TA100

Cell count (overnight culture): 2.09 x 10⁹CFU/mL

Concentration: (μg/plate)

Revertant colony number

Concentration: (μg/plate)

Revertant colony number

-S9

+S9

-S9

+S9

25000

22 SP, SR

37 SP, SR

19 SP, R

26 SP, SR

28 SP, SR

13 SP, SR

Untreated control

102

109

Mean

26.0

9.64

0.27

22.3

8.14

0.24

Mean

97.7

3.79

1.00

100.7

9.07

1.09

791

63 SR

88 SR

82 SR

97 SR

66 SR

86 SR

DMSO control

102

Mean

77.7

13.05

0.80

83.0

15.72

0.90

Mean

97.7

3.79

1.00

92.0

5.00

1.00

250

84 SR

81 SR

115 SR

106

100

Distilled water control*

103

109

Mean

93.3

18.82

0.96

97.7

9.71

1.06

Mean

98.0

4.58

1.00

98.0

9.54

1.07

79.1

113

109

101

106

Positive control

SAZ

(2 μg)

1196

1212

1184

2AA (2 μg)

2412

2388

2420

Mean

102.3

15.14

1.05

91.3

21.22

0.99

Mean

1197.3

14.05

12.22

2406.7

16.65

26.16

107

SP:Slight precipitate

R:Reduced background lawn

SR:Slightly reduced background lawn

MF:Mutation factor

SD:Standard deviation

+S9:with S9 mix

-S9:without S9 mix

Mean revertants (test item)

Mutation factor = --------------------------------------

Mean revertants (solvent control)

*Distilled water control was used because of SAZ

Mean

91.3

3.79

0.94

99.7

6.43

1.08

7.91

109

Mean

98.0

1.00

96.3

11.68

1.05

2.5

110

108

Mean

93.3

1.53

0.96

100.7

14.47

1.09

0.791

108

103

104

Mean

98.7

12.10

1.01

88.3

19.86

0.96

Confirmatory Mutation Test (Pre-Incubation Method)

Test Item: CUCURBIT[8]URIL

Strain: Salmonella typhimurium TA1535

Cell count (overnight culture): 2.67 x 10⁹CFU/mL

Concentration: (μg/plate)

Revertant colony number

Concentration: (μg/plate)

Revertant colony number

-S9

+S9

-S9

+S9

25000

15 SP, SR

16 SP, SR

10 SP, SR

5 P, SR

13 P, SR

11 P, SR

Untreated control

Mean

13.7

3.21

1.46

9.7

4.16

0.88

Mean

9.7

1.53

1.04

12.7

3.21

1.15

791

13 SR

12 SR

11 SR

15 SR

8 SR

DMSO control

Mean

12.3

0.58

1.32

11.3

3.51

1.03

Mean

9.3

2.08

1.00

11.0

1.00

250

14 SR

15 SR

14 SR

Distilled water control*

Mean

14.3

0.58

1.54

11.0

3.61

1.00

Mean

14.3

1.53

1.54

13.7

2.31

1.24

79.1

Positive control

SAZ

(2 μg)

1204

1176

1184

2AA (2 μg)

267

277

283

Mean

13.7

5.51

1.46

13.0

3.61

1.18

Mean

1188.0

14.42

82.88

275.7

8.08

25.06

P:Precipitate

SP:Slight precipitate

SR:Slightly reduced background lawn

MF:Mutation factor

SD:Standard deviation

+S9:with S9 mix

-S9:without S9 mix

Mean revertants (test item)

Mutation factor = --------------------------------------

Mean revertants (solvent control)

*Distilled water control was used because of SAZ

Mean

14.7

2.89

1.57

11.0

1.73

1.00

7.91

Mean

12.3

1.15

1.32

14.0

1.73

1.27

2.5

Mean

11.7

2.08

1.25

15.0

4.58

1.36

0.791

Mean

15.0

1.00

1.61

13.0

2.65

1.18

Confirmatory Mutation Test (Pre-Incubation Method)

Test Item: CUCURBIT[8]URIL

Strain: Salmonella typhimurium TA1537

Cell count (overnight culture): 2.26 x 10⁹CFU/mL

Concentration: (μg/plate)

Revertant colony number

Concentration: (μg/plate)

Revertant colony number

-S9

+S9

-S9

+S9

25000

1 P,R

2 P,R

0 P,R

1 P,R

2 P,R

Untreated control

Mean

1.3

0.58

0.19

1.0

1.00

0.14

Mean

8.7

2.08

1.24

5.3

2.08

0.76

791

3 SP,SR

5 SP,SR

6 SP,SR

5 SR

8 SR

11 SR

DMSO control

Mean

4.7

1.53

0.67

8.0

3.00

1.14

Mean

7.0

2.00

1.00

7.0

1.00

250

5 SR

8 SR

Positive control

9AA

(50 μg)

404

426

408

2AA

(2 μg)

198

204

220

Mean

6.0

1.73

0.86

10.7

6.03

1.52

Mean

412.7

11.72

58.95

207.3

11.37

29.62

79.1

P:Precipitate

SP:Slight precipitate

R:Reduced background lawn

SR:Slightly reduced background lawn

MF:Mutation factor

SD:Standard deviation

+S9:with S9 mix

-S9:without S9 mix

Mean revertants (test item)

Mutation factor = ------------------------------------

Mean revertants (solvent control)

Mean

7.3

1.53

1.05

9.0

4.36

1.29

Mean

10.7

3.51

1.52

11.3

2.52

1.62

7.91

Mean

8.0

2.65

1.14

9.7

2.52

1.38

2.5

Mean

9.0

7.00

1.29

11.7

1.53

1.67

0.791

Mean

8.0

3.00

1.14

10.3

0.58

1.48

Applicant's summary and conclusion

Conclusions:

Interpretation of results:
negative with and without metabolic activation

The reported data of this mutagenicity assay show that under the experimental conditions applied the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used. In conclusion, the test item CUCURBIT[8]URIL had no mutagenic activity in the examined bacterial strains under the test conditions of this study.

Executive summary:

The purpose of this study was to evaluate the mutagenic potential of the test item by measuring its ability to induce reverse mutations at selected loci of several strains of Salmonella typhimuriumand at the tryptophan locus of Escherichia coli WP2uvrA strain in the presence and absence of activated rat liver S9 fraction. Study was performed to – Ninth Addendum to OECD Guidelines for Testing of Chemicals, Section 4, No. 471, "Bacterial Reverse Mutation Test", 21 July 1997 – EPA Health Effects Test Guidelines, OPPTS 870.5100 "Bacterial Reverse Mutation Test", EPA

712-C-98-247, August 1998 – Commission Regulation (EC) No. 440/2008, B.13/14. "Mutagenicity: Reverse Mutation Test Using Bacteria", 30 May 2008 The method described in the above mentioned guidelines conforms to the guidelines for bacterial mutagenicity testing published by the major Japanese Regulatory Authorities including METI, MHLW and MAFF.

In conclusion The reported data of this mutagenicity assay show that under the experimental conditions applied the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.

In conclusion, the test item CUCURBIT[8]URIL had no mutagenic activity in the examined bacterial strains under the test conditions of this study.

Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.

Registration Dossier

Registration Dossier

Diss Factsheets

Genetic toxicity: in vitro

Administrative data

Data source

Reference

Materials and methods

Test guidelineopen all close all

Test guideline 1

Test guideline 2

Test guideline 3

Test material

Test material information

Constituent 1

Method

Species / strainopen all close all

Species / strain 1

Species / strain 2

Controls

Results and discussion

Test resultsopen all close all

Test results 1

Test results 2

Test results 3

Test results 4

Test results 5

Any other information on results incl. tables

Applicant's summary and conclusion

Registration Dossier

Registration Dossier

Diss Factsheets

Genetic toxicity: in vitro

Administrative data

Data source

Reference

Materials and methods

Test guidelineopen allclose all

Test material

Constituent 1

Method

Species / strainopen allclose all

Controls

Results and discussion

Test resultsopen allclose all

Any other information on results incl. tables

Applicant's summary and conclusion

Test guidelineopen all close all

Species / strainopen all close all

Test resultsopen all close all