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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
genetic toxicity in vitro, other
Type of information:
experimental study
Adequacy of study:
key study
Study period:
29 February 2016 TO 18 May 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
NON-CONFIDENTIAL NAME OF SUBSTANCE:
- Name of the substance on which testing is proposed to be carried out: Genamin DMG 75

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Version / remarks:
“In vitro Mammalian Chromosome Aberration Test” adopted on 26 September 2014
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
other: In vitro Chromosomal aberration test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Genamin DMG 75


Specific details on test material used for the study:
Test Item Name: Genamin DMG 75
Chemical Name ( IUPAC): N, N-Dimethyl-D-glucamine
CAS No.: 76326-99-3
Physical Appearance
(with colour) :Slightly yellowish liquid
Batch No.: RAK-KRS-00022
Purity (Declared by sponsor and/ or as per Certificate of Analysis) : 71.2%; (Active components ca. 75% in ca. 25% aqueous solution)
Batch Produced by: Global Amines Germany GmbH
84504 Burgkirchen, Germany
Date of Manufacture : April 2015
Date of Analysis : 24.09.2015
Date of Expiry/Valid up to : 01.04.2018
Storage Conditions: Ambient (21 to 29°C)

Method

Target gene:
Not applicable
Species / strain
Species / strain / cell type:
Chinese hamster Ovary (CHO)
Remarks:
CHO-K1
Details on mammalian cell type (if applicable):
CHO-K1 cell line procured from ATCC (American Type Culture Collection).
Additional strain / cell type characteristics:
other: Chromosome number of CHO-K1 is 2n=22.
Metabolic activation:
with and without
Metabolic activation system:
male Wistar Rats
Test concentrations with justification for top dose:
Based on the precipitation and pH test, 2 mg/mL was chosen as the highest dose for the initial cytotoxicity test. 2 mg/mL was chosen as the highest concentration for the chromosomal aberration test as the initial study was non toxic at 2 mg/mL
Vehicle / solvent:
Distilled Water- Vehicle(s) used: water]
- Justification for choice of vehicle: based on the results of solubility test
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
Distilled Water
Positive controls:
yes
Positive control substance:
cyclophosphamide
mitomycin C
Details on test system and experimental conditions:
CHO-K1 cell line procured from ATCC, Hams F12 medium containing 10% FBS
and 1 % penicillin-streptomycin incubated at 37°C with 5% CO2.
Rationale for test conditions:
As per guidelines
Evaluation criteria:
The cells were evaluated for structural aberrations in 150 metaphase plates for each replicate and the metaphases with aberrations.
Gaps were recorded separately.
Statistics:
Data (Percentage of cells with aberrations) was analyzed using SPSS Software version 22 for differences among vehicle control, positive control and test item groups using ANOVA following Dunnett’s test at a 95% level of confidence (p < 0.05) and the statistical significance was designated by the superscripts though out the report as stated below:
* Statistically significant (p < 0.05) change than the vehicle control group.

Results and discussion

Test results
Key result
Species / strain:
other: CHOK-1
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
not applicable
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No change
- Water solubility: 2mg/mL
- Precipitation: no precipitation at 2mg/mL

Remarks on result:
other: non-clastogenic

Any other information on results incl. tables

 

TABLE 1.           SUMMARY OF PERCENTAGE RICC FOR INITIAL CYTOTOXICITY TEST

Refer Appendix 2

Set No.

Treatment

Dose (mg/mL)

Initial Cell Count

(1×105 cells/flask)

Final Cell Count

(1×105cells/flask)

Final – initial cell count

(1×105cells/flask)

RICC

Reduction in RICC (%)

Set 1 (+S9)                  (3 to 6 hours)

Vehicle control

-

2.78

8.63

5.85

100

0

Test item

[Genamin DMG 75]

0.5

2.78

7.95

5.18

88.46

11.54

1

2.78

7.65

4.88

83.33

16.67

2

2.78

7.58

4.80

82.05

17.95

 

Set 2 (-S9)                     (3 to 6 hours)

Vehicle control

-

2.78

7.95

5.18

100.00

0

Test item

[Genamin DMG 75]

0.5

2.78

7.65

4.88

94.20

5.80

1

2.78

7.65

4.88

94.20

5.80

2

2.78

7.50

4.73

91.30

8.70

 

Set 3 (-S9)                   (18 to 20 hours)

Vehicle control

-

2.78

7.88

5.10

100.00

0

Test item

[Genamin DMG 75]

0.5

2.78

7.43

4.65

91.18

8.82

1

2.78

7.05

4.28

83.82

16.18

2

2.78

7.05

4.28

83.82

16.18

RICC:Relative increase in cell count, +S9: with metabolic activation, -S9: without metabolic activation.  

 

TABLE 2.     SUMMARY OFCHROMOSOMAL ABERRATIONSANDRICC

                                                                                              Refer Appendix3

Set No.

Treatment

Dose (mg/mL)

Initial Cell Count

(1×105cells/flask)

Final Cell Count

(1×105cells/flask)

Final – initial cell count

(1×105cells/flask)

RICC

Reduction in RICC (%)

Mean of Total Aberrations with Gaps

Mean of Total Aberrations without Gaps

Mean of Total Aberrated cells without Gaps

Percentage Mean of Aberrated Cells without Gaps

Set 1 (+S9) (3 to 6 hours)

Vehicle control

-

2.63

7.44

4.81

100

0

1.0

1.0

1.0

0.7

Test item

[Genamin DMG 75]

0.5

2.63

6.94

4.31

89.61

10.39

1.5

1.5

1.0

0.7

1

2.63

6.75

4.13

85.71

14.29

2.5

2.0

1.0

0.7

2

2.63

6.63

4.00

83.12

16.88

1.0

1.0

1.0

0.7

Positive Control

(Cyclophosphamide)

10 µg/mL

2.63

6.19

3.56

74.03

25.97

25.5

24.0

12.5

8.3*

RICC: Relative increase in cell count; *: Statistically significant;+S9: with metabolic activation.

 

 

 

 

  

 

TABLE 2 (Contd..,). SUMMARY OFCHROMOSOMAL ABERRATIONSAND RICC

                                                                                              Refer Appendix 3

Set No.

Treatment

Dose (mg/mL)

 

 

Initial Cell Count

(1×105cells/flask)

 

 

Final Cell Count

(1×105cells/flask)

Final – initial cell count

(1×105cells/flask)

RICC

Reduction in RICC (%)

Mean of Total Aberrations with Gaps

Mean of Total Aberrations without Gaps

Mean of Total Aberrated cells without Gaps

Percentage Mean of Aberrated Cells without Gaps

Set 2 (-S9) (3 to 6 hours)

Vehicle control

-

2.63

7.13

4.50

100.00

0.00

2.0

2.0

1.0

0.7

Test item

[Genamin DMG 75]

0.5

2.63

7.00

4.38

97.22

2.78

1.0

1.0

1.0

0.7

1

2.63

6.75

4.13

91.67

8.33

1.0

1.0

1.0

0.7

2

2.63

6.69

4.06

90.28

9.72

1.5

1.5

1.0

0.7

Positive Control

(Mitomycin-C)

0.05 µg/mL

2.63

6.44

3.81

84.72

15.28

21.5

19.5

13.5

9.0*

RICC: Relative increase in cell count; *: Statistically significant;-S9: without metabolic activation.

 

 

  

TABLE 2 (Contd..,). SUMMARY OF CHROMOSOMAL ABERRATIONS AND RICC

                                                                                              Refer Appendix 3

Set No.

Treatment

Dose (mg/mL)

Initial Cell Count

(1×105cells/flask)

Final Cell Count

(1×105cells/flask)

Final – initial cell count

(1×105cells/flask)

RICC

Reduction in RICC (%)

Mean of Total Aberrations with Gaps

Mean of Total Aberrations without Gaps

Mean of Total Aberrated cells without Gaps

Percentage Mean of Aberrated Cells without Gaps

Set 3 (-S9) (18 to 20 hours)

Vehicle control

-

2.63

7.19

4.56

100.00

0.00

1.0

1.0

1.0

0.7

Test item

[Genamin DMG 75]

0.5

2.63

7.06

4.44

97.26

2.74

1.0

1.0

1.0

0.7

1

2.63

6.69

4.06

89.04

10.96

1.0

1.0

1.0

0.7

2

2.63

6.50

3.88

84.93

15.07

2.0

2.0

1.0

0.7

Positive Control

(Mitomycin-C)

0.05 µg/mL

2.63

6.00

3.38

73.97

26.03

19.5

18.0

11.5

7.7*

RICC: Relative increase in cell count; *: Statistically significant;-S9: without metabolic activation.

 

 


 

 

 

 

 

 

 

 

 

 

 

 

 

 

Applicant's summary and conclusion

Conclusions:
The gentoxicity of the registration substance was investigated according to the Guideline OECD 473. The registration substance did not exhibit any mutagenic acitivity.
Executive summary:

The gentoxicity of the registration substance was investigated according to the Guideline OECD 476. The registration substance was incubated with Chinese hamster ovary cells and the cells were investigated for the increases of chromosome aberration. No increase of chromosome aberration was found. The registration substance did not exhibit any clastogenic activity.