Registration Dossier

Diss Factsheets

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The in-vitro genotoxicity of the registration substance, sodium N-lauroyl glutamate, is derived based on the read-across to sodium N-cocoyl glutamate and sodium N-cocoyl glycinate.

The registration substance and the read-across sources are amides of fatty acids and amino acids and can be characterized as "N-fatty acyl amino acids", of which endogeous occurence and metabolism are known. Based on the comparable chemical structure, comparable phys-chem data and expected comparable metabolism, these compound are likely exhibit comparable toxicity profiles.

 

The genotoxicity of sodium N-cocoyl glutamte was investigated in Ames test (OECD 471). No mutagenic activity was found.

The genotoxicity of sodium N-cocoyl glycinate was investigated in Ames test (OECD 471) and in mouse lymphoma assay (OECD 476). The results obtained in these studies are indicative of low mutagenic potential. Further, two chromosome aberrations studies are available reporting positive response. However, these

studies were re-evaluated according to the currently accepted Guideline OECD 473 (adopted in 2016) by the registrant and the data are evaluated as indicative of no significant clastogenicity.

 

Likewise, the registration substance, sodium N-lauroyl glutamate is considered as of no significant genotoxic activity.

 

Link to relevant study records

Referenceopen allclose all

Endpoint:
genetic toxicity in vitro, other
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
other information
Justification for type of information:
Justification is provided in the separate statement.
Reason / purpose for cross-reference:
assessment report
Reason / purpose for cross-reference:
assessment report
Reason / purpose for cross-reference:
assessment report
Reason / purpose for cross-reference:
assessment report
Reason / purpose for cross-reference:
assessment report
Conclusions:
The in-vitro genotoxicity of the registration substance sodium N-lauroyl glutamate is derived based on the read-across to Sodium N-cocoyl glutamate and Sodium N-cocoyl glycinate. No significant in-vitro genotoxicity can be reliably derived.
Executive summary:

The in-vitro genotoxicity of the registration substance, sodium N-lauroyl glutamate, is derived based on the read-across to sodium N-cocoyl glutamate and sodium N-cocoyl glycinate.

The registration substance and the read-across sources are amides of fatty acids and amino acids and can be characterized as "N-fatty acyl amino acids", of which endogeous occurence and metabolism are known. Based on the comparable chemical structure, comparable phys-chem data and expected comparable metabolism, these compound are likely exhibit comparable toxicity profiles.

 

The genotoxicity of sodium N-cocoyl glutamte was investigated in Ames test (OECD 471). No mutagenic activity was found.

The genotoxicity of sodium N-cocoyl glycinate was investigated in Ames test (OECD 471) and in mouse lymphoma assay (OECD 476). The results obtained in these studies are indicative of low mutagenic potential. Further, two chromosome aberrations studies are available reporting positive response. However, these studies were re-evaluated according to the currently accepted Guideline OECD 473 (adopted in 2016) by the registrant and the data are evaluated as indicative of no significant clastogenicity.

 

Likewise, the registration substance, sodium N-lauroyl glutamate is considered as of no significant genotoxic activity.

Endpoint:
in vitro gene mutation study in mammalian cells
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
Deviations:
no
Principles of method if other than guideline:
first experiment: 4 hours treatment with and without metabolic activation
second experiment: 24 hours treatment without metabolic activation, 4 hours treatment with metabolic activation
GLP compliance:
yes (incl. QA statement)
Type of assay:
mammalian cell gene mutation assay
Target gene:
Thymidine Kinase Locus
Species / strain / cell type:
mouse lymphoma L5178Y cells
Details on mammalian cell type (if applicable):
- Type and identity of media: RPMI
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: yes
- Periodically "cleansed" against high spontaneous background: yes
Additional strain / cell type characteristics:
other: Clone 3.7.2C
Metabolic activation:
with and without
Metabolic activation system:
Phenobarbital/Beta-Naphtoflavone induced Rat liver S9
Test concentrations with justification for top dose:
Experiment I:
without S9 mix: 5.9; 11.8; 23.5; 47.0; 94.0; 141.0 µg/mL
with S9 mix: 11.8; 23.5; 47.0; 94.0; 188.0; 282.0 µg/mL
Experiment II:
without S9 mix: 11.8; 23.5; 47.0; 94.0; 188.0; 282.0 µg/mL
with S9 mix: 8.8; 17.5; 35.0; 70.0; 105.0; 140.0 µg/mL
Following the expression phase of 48 hours the cultures at the maximum concentration of both main experiments were not continued due to exceedingly strong toxic effects.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: deionised water
- Justification for choice of solvent/vehicle: solubility properties
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
without metabolic activation
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
cyclophosphamide
Remarks:
with metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 4 hours with and without metabolic activation in experiment 1, 24 hours without metaoblic activation in experiment and 4 hours with metabolic activation in experiment 2
- Expression time (cells in growth medium): 48 hours
- Selection time (if incubation with a selection agent): 10 to 15 days

SELECTION AGENT (mutation assays): RPMI 1640 medium by addition of 5 µg/mL TFT

NUMBER OF REPLICATIONS: 2

NUMBER OF CELLS EVALUATED: >1,5 x 10 exp. 6 cells

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth


Evaluation criteria:
A test item is classified as mutagenic if the induced mutation frequency reproducibly exceeds a threshold of 126 colonies per 10 exp. 6 cells above the
corresponding solvent control or negative control, respectively.
A relevant increase of the mutation frequency should be dose-dependent.
A mutagenic response is considered to be reproducible if it occurs in both parallel cultures.
However, in the evaluation of the test results the historical variability of the mutation rates in negative
and/or vehicle con¬trols and the mutation rates of all negative and/or vehicle controls of this study are taken into consideration.
Results of test groups are generally rejected if the relative total growth, and the cloning efficiency 1 is less than 10 % of the vehicle control
unless the exception criteria specified by the IWGT recommendations are fulfilled.
Whenever a test item is considered mutagenic according to the above mentioned criteria, the ratio of small versus large colonies is used
to differentiate point mutations from clastogenic effects. If the increase of the mutation frequency is accompanied by a reproducible and
dose dependent shift in the ratio of small versus large colonies clastogenic effects are indicated.
Statistics:
Linear regression analysis (least squares) using SYSTAT(R) 11 (SYSTAT Software, Inc., 501, Canal Boulevard, Suite C, Richmond, CA 94804, USA)
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: Not effected
- Effects of osmolality: Not increased
- Evaporation from medium: Not examined
- Water solubility: Up to 25 %
- Precipitation: pre-experiment: at 750, 1500, 3000 µg/mL; main experiments: no precipitation occurred
- Other confounding effects: None


RANGE-FINDING/SCREENING STUDIES:
The highest concentration used in the pre-test was chosen with regard to the molecular weight of the test item. Test item concentrations between 23.4 and 3000 µg/mL were used to evaluate toxicity in the presence (4 h treatment) and absence (4 h and 24 h treatment) of metabolic activation.
Strong toxic effects occurred at 187.5 µg/mL and above with and without metabolic activation following 4 and 24 hours of treatment.
The test medium was checked for precipitation at the end of each treatment period (4 or 24 hours) before the test item was removed. Precipitation was observed by the unaided eye at 750 µg/mL and above with and without metabolic activation at both treatment intervals.
The dose range of the main experiments was set according to the data generated in the pre-experiment. The experimental part of the second
experiment with metabolic activation was terminated due to exceedingly severe cytotoxicity and repeated at lower concentrations. In both main
experiments the individual concentrations were generally spaced by a factor of 2.0 in the lower range. A narrower spacing was used at the highest concentrations to cover the cytotoxic range more closely.
To overcome problems with possible deviations in toxicity the main experiments were started with more than four concentrations.


Summary Table
      relative mutant   relative mutant  
  conc. µg S9 total colonies/   total colonies/  
  per mL mix growth 106cells threshold growth 106cells threshold
Column 1 2 3 4 5 6 7 8
Experiment I / 4 h treatment   culture I culture II
Solv. control with water - 100.0 100 226 100.0  71 197
Pos. control with MMS  19.5 -  40.3 292 226  28.4 330 197
Test item   5.9 - 116.4 109 226 100.2  79 197
Test item  11.8 - 102.1  88 226  90.2  81 197
Test item  23.5 - 114.9 101 226  90.3  71 197
Test item  47.0 -  94.7  89 226 104.3  60 197
Test item  94.0 -  22.4 111 226  14.9 123 197
Test item  141.0 - culture was not continued# culture was not continued#
       
Solv. control with water + 100.0  75 201 100.0 115 241
Pos. control with CPA   3.0 +  57.9 185 201  54.0 263 241
Pos. control with CPA   4.5  +   28.2 236 201  28.0 465 241
Test item  11.8  +   85.1  76 201  88.5 103 241
Test item  23.5  +   78.9  90 201  96.3  74 241
Test item  47.0  +   79.7  96 201 102.9  94 241
Test item  94.0  +   81.6  96 201  83.7  86 241
Test item  188.0  +   18.0 124 201  23.3 130 241
Test item  282.0  +  culture was not continued# culture was not continued#
Experiment II / 24 h treatment   culture I culture II
Solv. control with water - 100.0  67 193 100.0 129 255
Pos. control with MMS  13.0 -  20.4 649 193  25.9 697 255
Test item  11.8 -  59.5 117 193  61.8 169 255
Test item  23.5 -  66.8  84 193  74.8  94 255
Test item  47.0 -  64.5  77 193  52.2 128 255
Test item  94.0 -  43.7  69 193  39.1 140 255
Test item  188.0 -  1.6 195 193  1.6 171 255
Test item  282.0 - culture was not continued# culture was not continued#
Experiment II / 4 h treatment   culture I culture II
Solv. control with water + 100.0  70 196 100.0  98 224
Pos. control with CPA   3.0 +  44.6 218 196  26.4 408 224
Pos. control with CPA   4.5 +  22.0 272 196  16.4 378 224
Test item   8.8 +  47.1 163 196  82.9 125 224
Test item  17.5 +  74.4  87 196  77.8 133 224
Test item  35.0 +  48.6  86 196  92.9  95 224
Test item  70.0 +  50.5  99 196  98.6  78 224
Test item  105.0 +  47.4  67 196  81.7  80 224
Test item  140.0 + culture was not continued# culture was not continued#

threshold = number of mutant colonies per 106cells of each solvent control plus 126

#    culture not continued due to exceedingly strong toxic effects

 

Conclusions:
The genotoxicity of sodium N-cocoyl glycinate was investigated according to the Guideline OECD 476 (MLA). No mutagenic acitivty was found.
Executive summary:

The genotoxicity of sodium N-cocoyl glycinate was investigated according to the Guideline OECD 476 (MLA). In two independent experiments, the mouse lymphoma L5178Y cell cultures were treated with the test item up to the concentrations associated with 20% cell growth reduction with and without metabolic activation. No mutagenic acitivity was found.

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1994
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
Adopted May 26th 1983
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
S9 Mix, induced with Aroclor 1254
Test concentrations with justification for top dose:
Up to the highest dose as mentioned in the guideline
Vehicle / solvent:
Destilled water.
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
yes
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
other: 2-aminoanthracene
Details on test system and experimental conditions:
- The study included cytotoxicity preliminary assay using TA 100.
- On the day of the experiment the test substance was dissolved in Aqua bidest. at appropriate concentrations.
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Remarks:
tested up to the limit concentration
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Remarks:
tested up to the limit concentration
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Remarks:
tested up to the limit concentration
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Remarks:
tested up to the limit concentration
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
In the cytotoxicity test using TA 100, reduction of revertants was observed at 5000 µg/plate without metabolic activation.

Results obtained in preliminary cytotoxicity test

Strain /

status of metabolic activation

Concentration

[µg/plate]

No. of Revertants

 

 

Plate 1

Plate 2

Plate 3

Mean

SD

Ratio to vehicle control

TA 100, +S9

0 (vehicle control)

161

175

153

163.0

11.1

-

4

168

157

166

163.7

5.9

1.0

20

183

161

178

177.0

14.0

1.1

100

160

172

145

159.0

13.5

1.0

500

179

157

180

172.0

13.0

1.2

2500

150

141

176

155.7

18.2

1.0

5000

141

165

160

155.3

12.7

1.0

TA 100, -S9

0

120

176

177

157.7

32.6

-

4

134

154

131

139.7

12.5

0.9

20

142

160

158

153.3

9.9

1.0

100

156

146

139

147.0

8.5

0.9

500

163

171

149

157.7

16.7

1.0

2500

146

108

99

117.7

24.9

0.7

5000

67

68

56

63.7

6.7

0.4

Conclusions:
The genotoxicity of sodium N-cocoyl glutamate was investigated according to the Guideline OECD 471 (Ames test). No mutagenic activity was found.
Executive summary:

The genotoxicity of sodium N-cocoyl glutamte was investigated according to the Guideline OECD 471 (Ames test). No mutagenic activity was found.

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
adopted July 21, 1997
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
induced rat liver S9-mix
Test concentrations with justification for top dose:
3, 10; 33; 100; 333; 1000; 2500; and 5000 µg/plate / pre-experiment/experiment I and experiment II
Vehicle / solvent:
Deionised water
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
methylmethanesulfonate
other: 4-nitro-o-phenylene-diamine; 2-aminoanthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar plate incorporation; preincubation


DURATION
- Preincubation period: 1 hour
- Exposure duration: 72 hours


NUMBER OF REPLICATIONS: 3 plates


Evaluation criteria:
A test item is considered as a mutagen if a biologically relevant increase in the number of revertants exceeding the threshold of twice (strains TA 98, TA 100, and WP2 uvrA) or thrice (strains TA 1535 and TA 1537) the colony count of the corresponding solvent control is observed .
A dose dependent increase is considered biologically relevant if the threshold is exceeded at more than one concentration .
An increase exceeding the threshold at only one concentration is judged as biologically relevant if reproduced in an independent second experiment.
A dose dependent increase in the number of revertant colonies below the threshold is regarded as an indication of a mutagenic potential if reproduced in an independent second experiment. However, whenever the colony counts remain within the historical range of negative and solvent controls such an increase is not considered biologically relevant.
Statistics:
According to the OECD guideline 471, a statistical analysis of the data is not mandatory.
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:
valid
Positive controls validity:
valid
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:
valid
Positive controls validity:
valid
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:
valid
Positive controls validity:
valid
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:
valid
Positive controls validity:
valid
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:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS

Summary Tabellen

    Summary of Results Pre-Experiment and Experiment I

Study Name: 1211000

Study Code: Harlan CCR 1211000

Experiment: 1211000 VV Plate

Date Plated: 17/09/2008

Assay Conditions:

Date Counted: 24/09/2008

Metabolic

Activation

Test

Group

Dose Level

(µg/plate)

Revertant Colony Counts (Mean ±SD)

TA 1535

TA 1537

TA 98

TA 100

WP2 uvrA

Without Activation

Deionised water

20 ± 3

11 ± 3

35 ± 2

137 ± 5

55 ± 4

Untreated

18 ± 3

13 ± 3

37 ± 10

126 ± 4

47 ± 3

Test item

3 µg

20 ± 3

11 ± 4

29 ± 1

134 ± 12

50 ± 4

10 µg

19 ± 4

14 ± 2

33 ± 5

124 ± 8

52 ± 6

33 µg

18 ± 3

11 ± 4

35 ± 6

132 ± 10

55 ± 1

100 µg

19 ± 4

8 ± 0

37 ± 6

139 ± 16

52 ± 4

333 µg

13 ± 2

10 ± 4

32 ± 8

122 ± 15

63 ± 3

1000 µg

17 ± 2

7 ± 1

25 ± 5

84 ± 11 R

62 ± 2

2500 µg

8 ± 2 M R

2 ± 2 M R

19 ± 2 M R

36 ± 5 M R

59 ± 2

5000 µg

2 ± 2 M R

0 ± 1 M R

5 ± 2 M R

27 ± 3 M R

58 ± 2

NaN3

10 µg

2009 ± 31

2369 ± 114

4-NOPD

10 µg

424 ± 9

4-NOPD

50 µg

83 ± 3

MMS

3.0 µL

1017 ± 33

With Activation

Deionised water

23 ± 3

20 ± 5

37 ± 4

148 ± 5

68 ± 4

Untreated

24 ± 6

19 ± 7

42 ± 8

148 ± 14

64 ± 9

 Test item

3 µg

24 ± 3

19 ± 5

39 ± 5

129 ± 14

64 ± 4

10 µg

21 ± 5

17 ± 5

44 ± 8

135 ± 3

71 ± 2

33 µg

25 ± 3

19 ± 3

43 ± 6

132 ± 9

60 ± 11

100 µg

23 ± 1

15 ± 2

38 ± 4

134 ± 5

60 ± 4

333 µg

22 ± 5

13 ± 2

47 ± 6

136 ± 13

73 ± 11

1000 µg

15 ± 4

16 ± 3

35 ± 3

116 ± 13

69 ± 3

2500 µg

9 ± 3 M R

8 ± 1 M R

28 ± 2 M R

48 ± 7 M R

66 ± 1

5000 µg

5 ± 1 M R

1 ± 1 M R

13 ± 2 M R

34 ± 5 M R

59 ± 4

2-AA

2.5 µg

245 ± 16

191 ± 30

1267 ± 70

1506 ± 43

2-AA

10.0 µg

232 ± 24

Key to Positive Controls

Key to Plate Postfix Codes

NaN3

2-AA

4-NOPD

MMS

sodium azide

2-aminoanthracene

4-nitro-o-phenylene-diamine

methyl methane sulfonate

R

M

Reduced background growth

Manual count

     Summary of Results Experiment II

Study Name: 1211000

Study Code: Harlan CCR 1211000

Experiment: 1211000 HV2 Pre

Date Plated: 01/10/2008 / 14/10/2008*

Assay Conditions:

Date Counted: 08/10/2008 / 17/10/2008*

Metabolic

Activation

Test

Group

Dose Level

(µg/plate)

Revertant Colony Counts (Mean ±SD)

TA 1535

TA 1537

TA 98*

TA 100

WP2 uvrA

Without Activation

Deionised water

19 ± 6

12 ± 4

22 ± 2

131 ± 5

51 ± 8

Untreated

16 ± 6

10 ± 2

24 ± 4

137 ± 12

46 ± 8

 Test item

3 µg

17 ± 4

14 ± 5

23 ± 2

131 ± 8

55 ± 1

10 µg

18 ± 4

15 ± 2

24 ± 2

127 ± 9

52 ± 8

33 µg

18 ± 3

11 ± 6

24 ± 4

122 ± 1

55 ± 6

100 µg

22 ± 6

14 ± 3

23 ± 1

101 ± 15 R

56 ± 2

333 µg

12 ± 5

10 ± 4

15 ± 2

75 ± 7 R

47 ± 4

1000 µg

5 ± 2 R M

2 ± 3 M R

10 ± 1 M R

39 ± 5 M R

49 ± 5

2500 µg

2 ± 2 M R

0 ± 0 M R

3 ± 2 M R

31 ± 10 M R

46 ± 5

5000 µg

1 ± 1 M R

0 ± 0 M R

0 ± 0 M R

5 ± 2 M R

37 ± 7

NaN3

10 µg

1799 ± 67

1998 ± 31

4-NOPD

10 µg

1637 ± 116

4-NOPD

50 µg

105 ± 5

MMS

3.0 µL

418 ± 20

With Activation

Deionised water

17 ± 4

16 ± 5

26 ± 3

138 ± 9

58 ± 5

Untreated

18 ± 4

19 ± 7

29 ± 3

142 ± 37

64 ± 7

 Test item

3 µg

19 ± 4

17 ± 4

27 ± 0

131 ± 11

59 ± 3

10 µg

22 ± 4

17 ± 4

25 ± 2

132 ± 8

58 ± 3

33 µg

20 ± 2

19 ± 3

24 ± 2

141 ± 15

59 ± 6

100 µg

21 ± 1

15 ± 3

22 ± 5

140 ± 7

57 ± 5

333 µg

18 ± 4

14 ± 4

19 ± 3

110 ± 5 R

53 ± 2

1000 µg

11 ± 3 M R

12 ± 2 M R

10 ± 2 M R

74 ± 7 R

54 ± 1

2500 µg

9 ± 3 M R

8 ± 2 M R

4 ± 3 M R

31 ± 2 M R

59 ± 4

5000 µg

0 ± 0 M R

0 ± 0 M R

0 ± 0 M R

31 ± 4 M R

61 ± 3

2-AA

2.5 µg

256 ± 8

105 ± 10

1616 ± 151

1346 ± 157

2-AA

10.0 µg

334 ± 24

Key to Positive Controls

Key to Plate Postfix Codes

NaN3

2-AA

4-NOPD

MMS

sodium azide

2-aminoanthracene

4-nitro-o-phenylene-diamine

methyl methane sulfonate

R

M

Reduced background growth

Manual count

* Repeated experiment

Conclusions:
The genotoxicity of sodium N-cocoyl glycinate was investigated according to the Guideline OECD 471 (Ames test). No mutagenic acitivty was found.
Executive summary:

The genotoxicity of sodium N-cocoyl glycinate was investigated according to the Guideline OECD 471 (Ames test). No mutagenic activity was found.

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1998
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail
Reason / purpose for cross-reference:
read-across source
Qualifier:
equivalent or similar to guideline
Guideline:
other: See 'Remarks'
Version / remarks:
Guidelines for Toxicity Studies Required for Approval Application for Manufacturing (Importing) Pharmaceutical Products (Notification No. 24, Evaluation and Licensing Division I, September 11, 1989)
Principles of method if other than guideline:
The studa design is consisted of two experiments (reported as "direct method" and "metabolic activation method" in the given report. The first experiment ("direct method") comprises cell treatment of 24h/48h without metabolic activation. The second experiment ("metabolic activation method") comprises cell treatment of 6h with and wihout metabolic acitvation.
GLP compliance:
yes (incl. QA statement)
Type of assay:
in vitro mammalian chromosome aberration test
Species / strain / cell type:
other: Chinese hamster lung fibroblasts CHL
Details on mammalian cell type (if applicable):
- Type and identity of media: Eagle`s MEM supplemented with 10% calf serum
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: yes
Metabolic activation:
with and without
Metabolic activation system:
induced rat liver S9-mix
Test concentrations with justification for top dose:
Direct method:
40, 80, 120 and 160 microgram/mL (24 hour / 48 hour treatments)
Metabolic activation method:
78, 156, 313 and 625 microgram / mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Physiological saline
- Justification for choice of solvent/vehicle: Accepted solvent for this type of study
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
N-dimethylnitrosamine
mitomycin C
Details on test system and experimental conditions:
METHOD OF APPLICATION: Direct method (in medium) / Metabolic activation method (with and without S9-mix)

A] Direct method

DURATION
- Preincubation period: 3 days
- Exposure duration: 24 hours and 48 hours
- Fixation time (start of exposure up to fixation or harvest of cells): Following treatment (24 hour or 48 hour treatment)

SELECTION AGENT (mutation assays):
SPINDLE INHIBITOR (cytogenetic assays): Colcemid
STAIN (for cytogenetic assays): Giemsa`s staining solution in Soerensen buffer

NUMBER OF REPLICATIONS: duplicates

NUMBER OF CELLS EVALUATED: 200 in each concentration, 100 cells per dish

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index

B] Metabolic activation method

DURATION
- Preincubation period: 3 days
- Exposure duration: 6 hours (with and without S9-mix)
- Fixation time (start of exposure up to fixation or harvest of cells): Following treatment

SELECTION AGENT (mutation assays):
SPINDLE INHIBITOR (cytogenetic assays): Colcemid
STAIN (for cytogenetic assays): Giemsa`s staining solution in Soerensen buffer

NUMBER OF REPLICATIONS: duplicates

NUMBER OF CELLS EVALUATED: 200 in each concentration, 100 cells per dish

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index
Evaluation criteria:
Frequencies of cells with structural aberrations were calculated, both for the case in which dells with gaps were included, and in which cells with gaps were excluded. Assessments were based on the frequency of cells with structural aberrations including gaps.
Negative: < 5%
Equivocal: > 5%, < 10%
Positive: > 10%
Statistics:
No statistical analysis performed
Species / strain:
other: Chines hamster lung fibroblasts CHL
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
valid
Species / strain:
other: Chinese hamster lung fibroblasts CHL
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other:
Remarks:
The author of the study evaluated the results obtained as "positive". The registrant performed re-evaluation, taking account the cyotoxicity as the potential interferring factor for false-positive responce.

Results of cytotoxicity test

Concentrations (mg/mL)

Growth rate in the direct method (%)

Growth rate in the metabolic activation method (%)

 

Values in duplicates

Mean

 

Mean

0.010

94

96

95

96

101

99

0.020

90

90

90

102

98

100

0.039

79

73

76

89

89

89

0.078

72

78

74

86

83

85

0.156

15

14

15

89

91

90

0.313

0

0

0

68

50

59

0.625

0

0

0

36

35

36

1.25

0

0

0

0

0

0

2.5

0

0

0

0

0

0

5

0

0

0

0

0

0

Growth rate was calculated by regarding the growth rate in the solvent treatment group as 100%.

Approximate IC50:

     Direct method: 0.105 mg/mL

     Metabolic activation method: 0.420 mg/mL

Results of chromosome aberration test (direct method, 24h treatment)

Concentrations (µg/mL)

Number of observed cells

Number of cells with numerical aberrations

Number of cells with structural aberrations

g

ctb

cte

csb

cse

TA

TAG

Non-treatment control

200

0

0

0

0

0

0

0

0

Solvent control

200

0

0

0

0

0

0

0

0

40

200

1

0

1

1

o

0

2

2

80

200

0

0

0

0

0

0

0

0

120

200

0

0

0

0

0

0

0

0

160

200

0

0

0

0

0

0

0

0

Positive control

200

0

0

30

69

0

2

91

91

g: chromatid gap or chromosome gap

ctb: chromatid break

cte: chromatid exchange

csb: chromosome break

cse: chromosome exchange

TA: total number of aberration cells excluding gaps

TAG: total number of aberration cells including gaps

Solvent control: physiological salne

Positive control: MMC (0.05 µg/mL)

Results of chromosome aberration test (direct method, 48h treatment)

Concentrations (µg/mL)

Number of observed cells

Number of cells with numerical aberrations

Number of cells with structural aberrations

g

ctb

cte

csb

cse

TA

TAG

Non-treatment control

200

1

0

1

0

0

0

1

1

Solvent control

200

0

0

0

0

0

0

0

0

40

200

5

0

0

1

0

0

1

1

80

200

1

0

2

2

0

0

4

4

120

200

1

0

3

0

0

0

3

3

Positive control

200

0

1

50

92

0

7

128

128

g: chromatid gap or chromosome gap

ctb: chromatid break

cte: chromatid exchange

csb: chromosome break

cse: chromosome exchange

TA: total number of aberration cells excluding gaps

TAG: total number of aberration cells including gaps

Solvent control: physiological salne

Positive control: MMC (0.05 µg/mL)

Results of chromosome aberration test (metabolic activation method, without S9 mix)

Concentrations (µg/mL)

Number of observed cells

Number of cells with numerical aberrations

Number of cells with structural aberrations

g

ctb

cte

csb

cse

TA

TAG

Non-treatment control

200

1

0

0

1

0

0

1

1

Solvent control

200

0

0

2

0

0

0

2

2

78

200

2

0

0

1

0

0

1

1

156

200

0

0

0

1

0

0

1

1

313

200

4

0

0

0

0

0

0

0

625

200

2

0

1

0

0

0

1

1

Positive control

200

1

1

1

0

0

0

1

2

g: chromatid gap or chromosome gap

ctb: chromatid break

cte: chromatid exchange

csb: chromosome break

cse: chromosome exchange

TA: total number of aberration cells excluding gaps

TAG: total number of aberration cells including gaps

Solvent control: physiological salne

Positive control: N-nitrosodimethylamine (0.4 µg/mL)

Results of chromosome aberration test (metabolic activation method, with S9 mix)

Concentrations (µg/mL)

Number of observed cells

Number of cells with numerical aberrations

Number of cells with structural aberrations

g

ctb

cte

csb

cse

TA

TAG

Non-treatment control

200

1

1

1

0

0

0

1

2

Solvent control

200

3

1

2

0

0

0

2

2

78

200

0

0

2

0

0

0

2

2

156

200

2

0

0

1

0

0

1

1

313

200

3

0

0

0

0

0

0

0

625

200

0

0

11

35

0

3

46

46

Positive control

200

0

1

45

126

0

7

146

147

g: chromatid gap or chromosome gap

ctb: chromatid break

cte: chromatid exchange

csb: chromosome break

cse: chromosome exchange

TA: total number of aberration cells excluding gaps

TAG: total number of aberration cells including gaps

Solvent control: physiological salne

Positive control: N-nitrosodimethylamine (0.4 µg/mL)

Comments of the registrant: the concentration of 625 µg/mL corresponds to the 35% of cell growth when compared to control and therefore not compliant to the OECD 473 Guideline adopted in 2016. Discarding the result obtained at this concentration, the overall result of the study should be negative.

Conclusions:
The genotoxicity of N-cocoyl glycinate was investigated in in-vitro chromosome aberration test. No genotoxic activity was at concentrations associated cyotoxicity of cell growth reduction less than 50%.
Executive summary:

The genotoxicity of N-cocoyl glycinate was investigated in in-vitro chromosome aberration test. Chines hamster lung fibroblasts cell cultures were used as test system in two independent experiments.

In the first experiments the cells were treated for 24h or 48h without metabolic acitivation. The highest concentrations used corresponded to the cell growth reduction down to 15% of control.

In the second experiment the cells were treated for 6h with or without metabolic activation. The highest concentrations used corresponded to the cell growth reduction down to 36% of control

No increase of aberration frequency was observed without metabolic activation in the first and in the second experiment. Significant increase of aberration frequency was obtained with metabolic activation at the highest concentration applied. The author of the study evaluated the results obtained as "positive".

The registrant performed re-evaluation in 2018, taking account the cyototoxicity in the culture medium as potential interfering factor responsible for a false-positve response (according to the current OECD Guideline, version 2016). Based on the re-evaluation, no significant genotoxic activity can be assigned up to the insoluble concentrations for N-cocoyl glycinate.

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1992
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
The concentration used for the test was selected only based on the cytotoxicity, but not based on the precipitation.The results obtained at concentrations with precipiations are not considered as valid.
Reason / purpose for cross-reference:
assessment report
Qualifier:
according to guideline
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Version / remarks:
adpted July 21, 1997
GLP compliance:
yes (incl. QA statement)
Type of assay:
other: in-vitro chromosome aberration
Species / strain / cell type:
lymphocytes:
Metabolic activation:
with and without
Metabolic activation system:
rat liver S9
Test concentrations with justification for top dose:
The highest dose selected was the dose causing 50% reduction of mitotic indices of control.
Vehicle / solvent:
water
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
cyclophosphamide
ethylmethanesulphonate
Details on test system and experimental conditions:
METHOD OF APPLICATION: in culture medium. The chromosomes were prepared 22 hours after start of treatment with the test item. Evaluation of two cultures per dose group.
DURATION
- Exposure duration: 4 hours (+/- S9 mix) and 22 hours (- S9 mix)
- Fixation time (start of exposure up to fixation or harvest of cells): 22 hours
SPINDLE INHIBITOR (cytogenetic assays): Colcemid
STAIN (for cytogenetic assays): Giemsa
NUMBER OF REPLICATIONS: about 1.5
NUMBER OF CELLS EVALUATED: At least 100 per culture

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index
Species / strain:
lymphocytes:
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
valid
Species / strain:
lymphocytes:
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: The author of the study evaluated the results obtained as "positive". The registrant performed re-evaluation, taking account the precipitation in the culture medium as the potential artifical factor for false-positive responce.

Summary of results of the chromosome aberration study: without metabolic activation

Exp

Preparation Interval

Test item concentration

[µg/mL]

Mitotic indices in % of control

Aberrant cells

Carrying exchange

Evaluation

based on the Guideline

adopted July 29, 2016

Incl. gaps*

Excl. gaps*

Exposure period 4 hrs without S9 mix

IA

22 hrs

Deionized water,

10% (v/v)

100.0

1.5

1.5

0.0

Negative:

Precipitation occurred at 99.5 µg/mL, which should be considered as the valid high dose.

The dose 304.6 µg/mL is not compliant to the Guideline adopted in 2016.

 

 

EMS,

770 µg/mL

51.7

8.5

8.5S

0.5

 

 

56.8

82.1

0.0

0.0

0.0

 

 

99.5P

70.3

0.0

0.0

0.0

 

 

304.6P

61.8

1.0

1.0

0.0

IB

22 hrs

Deionized water,

10% (v/v)

100.0

0.5

0.5

0.0

Negative:

Precipitation occurred at 50.0 µg/mL.

The doses 200 and 325 µg/mLis not compliant to the Guideline adopted in 2016.

 

 

EMS,

770 µg/mL

74.8

8.5

8.5S

3.0

 

 

25.0

101.0

1.5

0.5

0.0

 

 

50.0P

98.0

0.5

0.5

0.0

 

 

200.0P

83.0

2.0

2.0

0.0

 

 

325.0P

45.1

2.5

2.5

0.0

Exposure period 22 hrs without S9 mix

IIA

22 hrs

Deionized water,

10% (v/v)

100.0

0.0

0.0

0.0

Negative:

No Precipitation observed, but the highest dose induced clear cytotoxicity. 

 

 

 

EMS,

660 µg/mL

38.0

21.5

20.5S

9.0

 

 

30.5

96.1

1.0

1.0

0.0

 

 

53.3

82.8

1.0

1.0

0.0

 

 

93.3

62.4

1.0

1.0

0.0

IIB

22 hrs

Deionized water,

10% (v/v)

100.0

2.5

2.0

0.0

Negative:

No Precipitation observed, but the highest dose induced clear cytotoxicity. 

 

 

 

EMS,

550 µg/mL

43.1

22.0

21.5S

9.0

 

 

100.0

86.3

3.0

2.0

0.0

 

 

140.0

56.5

1.5

1.5

0.0

 

 

160.0

37.8

0.5

0.5

0.0

*  Including cells carrying exchanges

P  Precipitation occurred at the end of treatment

S  Aberration frequency statistically significant higher than corresponding control values

Summary of results of the chromosome aberration study: with metabolic activation

Exp

Preparation Interval

Test item concentration

[µg/mL]

Mitotic indices in % of control

Aberrant cells

Carrying exchange

Evaluation

based on the Guideline

adopted July 29, 2016

Incl. gaps*

Excl. gaps*

Exposure period 4 hrs with S9 mix

IA

22 hrs

Deionised water 10.0 % (v/v)

100.0

1.0

1.0

0.0

Negative:

Precipitation occurred at 99.5 µg/mL, which should be considered as the valid high dose.

The dose 304.6 µg/mL is not compliant to the Guideline adopted in 2016.

 

 

CPA,

15.0 µg/mL

79.6

12.5

12.0S

1.0

 

 

56.8

100.0

0.0

0.0

0.0

 

 

99.5P

98.9

0.5

0.5

0.0

 

 

304.6P

88.9

1.0

1.0

0.0

IIA

22 hrs

Deionised water 10.0 % (v/v)

100.0

2.0

2.0

0.0

Negative:

Precipitation occurred at 100 µg/mL.

The dose 500 µg/mL is not compliant to the Guideline adopted in 2016.

 

 

CPA,

15.0 µg/mL

69.9

22.5

22.0S

5.5

 

 

50.0

89.3

1.0

1.0

0.0

 

 

100.0P

93.2

2.0

2.0

0.0

 

 

500.0P

87.4

3.5

3.5

1.0

IIB

22 hrs

Deionized water,

10% (v/v)

100.0

1.5

1.5

0.0

Negative:

Precipitation occurred at 100 µg/mL.

The dose 520 µg/mL is not compliant to the Guideline adopted in 2016.

 

 

CPA,

7.5 µg/mL

78.2

9.5

8.0S

2.0

 

 

50.0

72.8

2.0

1.5

0.0

 

 

100.0P

87.9

1.5

1.5

0.0

 

 

520.0P

56.8

8.0

8.0S

2.5

IIC

22 hrs

Deionized water,

10% (v/v)

100.0

0.5

0.5

0.0

Negative:

Precipitation occurred at 100 µg/mL, and the aberration cells were statistically increased. Taking account the results obtained in other three independent experiments were negative and the incidence is marginally increased, the overall assessment should be negative.

The doses 400 and 440 µg/mL are not compliant to the Guideline adopted in 2016.

 

 

CPA,

7.5 µg/mL

54.9

19.5

19.5S

6.0

 

 

50.0

91.7

2.0

1.5

0.0

 

 

100.0P#

99.5

2.8

2.5S

0.3

 

 

400.0P#

61.1

6.3

6.0S

1.0

 

 

440.0P#

37.8

9.0

8.8S

1.0

*  Including cells carrying exchanges

P  Precipitation occurred at the end of treatment

S  Aberration frequency statistically significant higher than corresponding control values

#   Evaluation of 200 metaphases per culture

Conclusions:
The genotoxicity of N-cocoyl glycinate was investigated according to the Guideline OECD 473. No genotoxic activity was observed up to the insoluble concentrations.
Executive summary:

The genotoxicity of N-cocoyl glycinate was investigated according to the Guideline OECD 473 (version 1997). Human lymphocytes were treated with the test substance with and without metabolic activation up to the concentration associated with 50% reduction of mitotic indices. In general, reduced cytotoxicity was observed with metabolic activation, leading to the use of higher concentrations with metabolic activation for the genotoxicity investigation. These concentrations were associated with enhanced precipitation.

Total five independent experiments were performed. No increase of aberration frequency was observed without metabolic activation. Significant increase of aberration frequency was obtained with metabolic activation at concentrations associated not only with significant cytotoxicity but also with enhanced precipitation. The author of the study evaluated the results obtained as "positive".

The registrant performed re-evaluation in 2018, taking account the precipitation in the culture medium as potential interfering factor responsible for a false-positve response (according to the current OECD Guideline, version 2016). Based on the re-evaluation, no significant genotoxic activity can be assigned up to the insoluble concentrations for N-cocoyl glycinate.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

In-vivo genotoxicity

The in-vivo genotoxicity of the registration substance, sodium N-lauroyl glutamate, is derived based on the read-across to sodium N-cocoyl glycinate.

The registration substance and the read-across sources are amides of fatty acids and amino acids and can be characterized as "N-fatty acyl amino acids", of which endogeous occurence and metabolism are known. Based on the comparable chemical structure, comparable phys-chem data and expected comparable metabolism, these compound are likely exhibit comparable toxicity profiles.

 

The in-vivo clastogenicity of sodium N-cocoyl glycinate was investigated in micronucleus study in bone marrow in mice. Male mice were treated intraperitoneally at concentrations of 400, 200, 100 and 50 mg/kg bw, twice with a 24 hours interval. One death was observed in the 200 mg/kg body weight group (1/6 cases), and four deaths were observed in the 400 mg/kg body weight group (4/6 cases). The surviving animals were killed at 24 hours after the final treatment and the bone-marrow smears were prepared. The proportion of polychromatic erythrocytes (PCE) to total erythrocytes was lower in the 100 and 200 mg/kg bw groups, indicating toxic effect in the bone marrow tissue. The percentage of micronucleated polychromatic ethythrocytes (MNPCE) in the treated group was not significantly higher than that in the negative control group.

Sodium N-cocoyl glycinate is not clastogenic in in-vivo test system.

 

Likewise, the registration substance, sodium N-lauroyl glutamate is considered as of no significant clstogenic activity under in-vivo condition.

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
other information
Justification for type of information:
Justification is provided in the separate statement.
Reason / purpose for cross-reference:
assessment report
Conclusions:
The in-vivo genotoxicity of the registration substance sodium N-lauroyl glutamate is derived based on the read-across to Sodium N-cocoyl glycinate. Sodium N-cocoyl glycinate was found to be not clastogenic in micronucleus study in bone marrow in mice. No significant in-vivo clastogenicity can be derived for the registration substance sodium N-lauroyl glutamte.
Executive summary:

The in-vivo genotoxicity of the registration substance, sodium N-lauroyl glutamate, is derived based on the read-across to sodium N-cocoyl glycinate.

The registration substance and the read-across sources are amides of fatty acids and amino acids and can be characterized as "N-fatty acyl amino acids", of which endogeous occurence and metabolism are known. Based on the comparable chemical structure, comparable phys-chem data and expected comparable metabolism, these compound are likely exhibit comparable toxicity profiles.

 

The in-vivo clastogenicity of sodium N-cocoyl glycinate was investigated in micronucleus study in bone marrow in mice. Male mice were treated intraperitoneally at concentrations of 400, 200, 100 and 50 mg/kg bw, twice with a 24 hours interval. One death was observed in the 200 mg/kg body weight group (1/6 cases), and four deaths were observed in the 400 mg/kg body weight group (4/6 cases). The surviving animals were killed at 24 hours after the final treatment and the bone-marrow smears were prepared. The proportion of polychromatic erythrocytes (PCE) to total erythrocytes was lower in the 100 and 200 mg/kg bw groups, indicating toxic effect in the bone marrow tissue. The percentage of micronucleated polychromatic ethythrocytes (MNPCE) in the treated group was not significantly higher than that in the negative control group.

Sodium N-cocoyl glycinate is not clastogenic in in-vivo test system.

 

Likewise, the registration substance, sodium N-lauroyl glutamate is considered as of no significant clstogenic activity.

Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1998
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail
Reason / purpose for cross-reference:
read-across source
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
micronucleus assay
Species:
mouse
Strain:
ICR
Sex:
male
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Japan
- Age at study initiation: approximately 8 weeks
- Weight at study initiation: 33.2 - 40.6 g
- Assigned to test groups randomly: yes
- Housing: aluminium box-type cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 26°C
- Humidity (%): 40 - 70 %
- Air changes (per hr): 10 - 15 cycles per hour
- Photoperiod (hrs dark / hrs light): 12 hour light / dark cycle
Route of administration:
intraperitoneal
Vehicle:
- Vehicle(s)/solvent(s) used: physiological saline
- Justification for choice of solvent/vehicle: recommended vehicle
- Concentration of test material in vehicle: 0.5, 1, 2 and 4 % (w/v)

Duration of treatment / exposure:
24 hours
Frequency of treatment:
two intraperitoneal injections within a 24 hour interval
Post exposure period:
no post exposure period
Dose / conc.:
400 mg/kg bw/day
Dose / conc.:
200 mg/kg bw/day
Dose / conc.:
100 mg/kg bw/day
Dose / conc.:
50 mg/kg bw/day
No. of animals per sex per dose:
6
Control animals:
yes, concurrent vehicle
Positive control(s):
Mitomycin C
Tissues and cell types examined:
bone marrow from right femur
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION:
Doses were selected on the basis of a preliminary dose-range finder.

TREATMENT AND SAMPLING TIMES:
Treatment period was 24 hours. The test solutions were administered intraperitoneally, twice within this 24 hour time interval.

DETAILS OF SLIDE PREPARATION:


METHOD OF ANALYSIS:

OTHER:
Sex:
male
Genotoxicity:
negative
Toxicity:
yes
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
RESULTS OF RANGE-FINDING STUDY
- Dose range: 250, 500, 1000, 2000 mg/kg body weight
- Solubility: yes
- Clinical signs of toxicity in test animals: yes


RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): no induction of micronuclei
- Appropriateness of dose levels and route: dose levels based on pre-test, intraperitoneal administration is a recommended route
- Statistical evaluation: yes

Results of micronucleus test in male CD-1 mice after introperitoneal administration of sodium N-cocoyl glycinate

Concentrations (mg/mL)

No. of animals survived / 

No, of animals treated

Frequency of MNPCE (%)

PCE/RBC

Solvent control

6/6

0.07 ± 0.08

58.8 ± 2.68

50 x 2

6/6

0.18 ± 0.10

59.5 ± 7.45

100 x 2

6/6

0.07 ± 0.10

** 48.8 ± 5.83

200 x 2

5/6

0.08 ± 0.11

** 36.8 ± 4.61

400 x 2

2/6

0.10 

 * 50.6 ± 2.69

Positive control (MMC)

6/6

4.77 ± 1.02

* 50.6 ± 2.69

PCE: polychromatic erythrocytes

RBC: total erythrocytes

MNPCE: micronucleated PCE

MMC: mitomycin C

* significantly different from solvent control (P< 0.05)

** significantly different from solvent control (P < 0.01)

Conclusions:
The in-vivo clastogenicity of sodium N-cocoyl glycinate was investigated in micronucleus study in bone marrow in mice. No significant clastogenicity was found.
Executive summary:

The in-vivo clastogenicity of sodium N-cocoyl glycinate was investigated in micronucleus study in bone marrow in mice. Male mice were treated intraperitoneally at concentrations of 400, 200, 100 and 50 mg/kg bw, twice with a 24 hours interval. One death was observed in the 200 mg/kg body weight group (1/6 cases), and four deaths were observed in the 400 mg/kg body weight group (4/6 cases). The surviving animals were killed at 24 hours after the final treatment and the bone-marrow smears were prepared. The proportion of polychromatic erythrocytes (PCE) to total erythrocytes was lower in the 100 and 200 mg/kg bw groups, indicating toxic effect in the bone marrow tissue. The percentage of micronucleated polychromatic ethythrocytes (MNPCE) in the treated group was not significantly higher than that in the negative control group.

Sodium N-cocoyl glycinate is not clastogenic in in-vivo test system.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Justification for classification or non-classification

The genotoxicity property of the registration substance sodium N-lauroyl glutamate is derived based on the read-across to Sodium N-cocoyl glutamate and Sodium N-cocoyl glycinate. One Ames test is available for sodium N-cocoyl glutamate and one Ames test, two chromosome aberration tests and one mouse lymphoma assay are available for sodium N-cocoyl glycinate. Further one micronucleus test in bone marrow of mice is available for sodium N-cocoyl glycinate. No significant genotoxic activity was found in all available studies.

No significant genotoxicity property can be reliably derived for the registration substance and therefore no classification is assigned.