Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

Administrative data

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
December 9, 2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2005
Report Date:
2005

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Qualifier:
according to
Guideline:
EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
GLP compliance:
yes
Type of assay:
in vitro mammalian chromosome aberration test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): NLP #8 D-Glucitol, propoxylated
- molecular weight: (Mn = approx. 600 g/mol)

- Analytical purity: > 99%
- Lot/batch No.: 304N0

Method

Target gene:
Induction of chromosomal damage (clastogenicity).
Species / strain
Species / strain / cell type:
lymphocytes:
Details on mammalian cell type (if applicable):
human
Metabolic activation:
with and without
Metabolic activation system:
rat liver homogenated (S9) with standard co-facotrs
Test concentrations with justification for top dose:
0, 625, 1250, 2500, 5000 (µg/ml)
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes

Results and discussion

Test results
Species / strain:
lymphocytes: human
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Remarks:
> 5000 µg/ml
Vehicle controls validity:
valid
Untreated negative controls validity:
valid

Any other information on results incl. tables

*** Denotes statistical significant different (P<0.001)

Table 1: Results Mitotic Index - Experiment 1:

Dose level 4 hours treatment without S9 4 hours treatment with S9  
µg/ml A B mean % of control A B mean % of control  
0 6.15 4.55 5.35 100 3.50 6.15 4.83 100  
625 - - - - - - - -  
1250 8.55 5.34 6.95 130 5.95 5.65 6.80 120  
2500 6.05 7.65 6.85 128 6.40 5.40 5.90 122  
5000 8.25 4.00 6.13 115 4.80 5.10 4.95 102  
MMC 0.4 2.15 2.25 2.20 41 NA NA NA NA  
CP 7.5 NA NA NA NA 2.15 3.85 3 62  

Table 2: Results Mitotic Index - Experiment 2:

Dose level 4 hours treatment without S9 4 hours treatment with S9
µg/ml A B mean % of control A B mean % of control
0 5.05 6.45 5.75 100 4.55 4.80 4.63 100
625 - - - - - - - -
1250 5.85 6.15 6.00 104 4.60 5.35 4.98 108
2500 3.85 4.30 4.08 71 4.30 3.00 3.65 79
5000 5.45 3.15 4.30 75 5.45 4.30 4.88 105
MMC 0.4 1.95 0.30 1.13 20 NA NA NA NA
CP 7.5 NA NA NA NA 0.65 0.75 0.70 15

Table 3: Results of the Chromosome Aberration Test - experiment 1 without metabolic Activation:

Treatment group [µg/ml] Replicate Mitotic Index Number of cells Scored Number of aberrations Total Number of Aberrations Frequency of Aberrant Cells [%]
Gaps Chromatid Chromosome
Breaks Exchanges Breaks Exchanges Others +Gaps -Gaps +Gaps -Gaps
Vehicle control A 6.15 100 0 0 0 0 0 0 0 0 0 0
B 4.55 100 0 0 0 0 0 0 0 0 0 0
total 200 0 0 0 0 0 0 0 0 0 0
(100) (0.0) (0.0)
1250 A 8.55 100 1 0 0 0 0 0 1 0 1 0
B 5.35 100 0 0 0 0 0 0 0 0 0 0
total 200 1 0 0 0 0 0 1 0 1 0
(130) (0.5) (0.0)
2500 A 6.05 100 1 0 0 0 1 0 2 1 2 1
B 7.65 100 1 0 0 0 0 0 1 0 1 0
total 200 2 0 0 0 1 0 3 1 3 1
(128) (1.5) (0.5)
5000 A 8.25 100 2 0 0 0 0 0 2 0 2 0
B 4.00 100 1 0 0 1 0 0 2 1 2 1
total 200 3 0 0 1 0 0 4 1 4 1
(115) (2.0) (0.5)
positive control A 2.15 50 4 16 22 1 0 0 43 39 27 25
B 2.25 100 7 13 26 2 0 48 41 29 25
total 150 11 29 48 3 0 0 91 80 56 50***
(41) (37.3) (33.3)

Table 4: Results of Chromosome Abberation test Experiment 1 with metabolic activation (S9)

Treatment group [µg/ml] Replicate Mitotic Index Number of cells Scored Number of aberrations Total Number of Aberrations Frequency of Aberrant Cells [%]
Gaps Chromatid Chromosome
Breaks Exchanges Breaks Exchanges Others +Gaps -Gaps +Gaps -Gaps
Vehicle control A 3.50 100 1 0 0 0 0 0 1 0 1 0
B 6.15 100 2 0 0 0 0 0 2 0 2 0
total 200 3 0 0 0 0 0 3 0 3 0
(100) (1.5) (0.0)
1250 A 5.95 100 1 0 0 0 0 0 1 0 1 0
B 5.65 100 0 0 0 0 0 0 0 0 0 0
total 200 1 0 0 0 0 0 1 0 1 0
(120) (0.5) (0.0)
2500 A 6.40 100 0 0 0 0 0 0 0 0 0 0
B 100 0 0 0 0 0 0 0 0 0 0
total 200 0 0 0 0 0 0 0 0 0
(122) (0.0) (0.0)
5000 A 4.80 100 0 0 0 0 0 0 0 0 0 0
B 5.10 100 1 1 0 0 0 0 2 1 2 1
total 200 1 1 0 0 0 0 2 1 2 1
(102) (1.0) (0.5)
positive control A 2.15 100 6 30 9 4 0 0 49 43 37 35
B 3.85 100 3 5 1 8 0 0 17 14 8 6
total 200 9 35 10 12 0 0 66 57 45 41***
(22.5) (20.5)

Table 5:Results of the Chromosome Aberration Test - Experiment 2 without metabolic Activation:

Treatment group [µg/ml] Replicate Mitotic Index Number of cells Scored Number of aberrations Total Number of Aberrations Frequency of Aberrant Cells [%]
Gaps Chromatid Chromosome
Breaks Exchanges Breaks Exchanges Others +Gaps -Gaps +Gaps -Gaps
Vehicle control A 5.05 100 0 0 0 0 0 0 0 0 0 0
B 6.45 100 0 0 0 0 0 0 0 0 0 0
total 200 0 0 0 0 0 0 0 0 0 0
(100) (0.0) (0.0)
1250 A 5.85 100 0 0 1 0 0 0 1 1 1 1
B 6.15 100 0 0 0 0 0 0 0 0 0 0
total 200 0 0 1 0 0 0 1 1 1 1
(0.5) (0.5)
2500 A 3.85 100 0 0 0 1 0 0 1 1 1 1
B 4.30 100 1 0 0 0 0 0 1 0 1 0
total 200 1 0 0 1 0 0 2 1 2 1
(1.0) (0.5)
5000 A 5.45 100 0 0 0 0 0 0 0 0 0 0
B 3.15 100 0 0 0 0 0 0 0 0 0 0
total 200 0 0 0 0 0 0 0 0 0
(75) (0.0) (0.0)
positive control A 1.95 100 9 15 30 2 0 0 55 46 33 30
B 0.30 39 5 14 18 1 0 0 38 33 19 17
total 139 14 28 48 3 0 0 93 79 52 47***
(20) (37.4) (33.8)

Table 6:Results of the Chromosome Aberration Test - Experiment 2 with metabolic Activation:

Replicate Mitotic Index Number of cells Scored Number of aberrations Total Number of Aberrations Frequency of Aberrant Cells [%]
Gaps Chromatid Chromosome
Breaks Exchanges Breaks Exchanges Others +Gaps -Gaps +Gaps -Gaps
A 4.55 100 1 0 0 0 0 0 1 0 1 0
B 4.70 100 0 0 0 0 0 0 0 0 0
total 200 1 0 0 0 0 0 1 0 1 0
(100) (0.5) (0.0)
A 4.60 100 0 0 0 0 0 0 0 0 0 0
B 5.35 100 0 0 0 0 0 0 0 0 0 0
total 200 0 0 0 0 0 0 0 0 0
(108) (0.0) (0.0)
A 4.30 100 3 0 0 0 1 0 3 1 3 1
B 3.00 100 1 0 1 0 0 0 2 1 2 1
total 200 3 0 1 0 1 0 5 2 5 2
(79) (2.5) (1.0)
A 5.45 100 2 0 0 0 0 0 2 0 2 0
B 4.30 100 1 1 1 0 0 0 3 2 3 2
total 200 3 1 1 0 0 0 5 2 5 2
(105) (2.5) (1.0)
A 0.65 100 11 13 4 1 0 0 29 18 20 13
B 0.75 100 12 15 8 5 0 0 40 28 33 22
total 200 23 28 12 6 0 0 69 46 53 35***
(15) (26.5) (17.5)

Table 7: Mean Frequency of Polyploid Cells - Experiment 1:

Dose [µg/ml] with S9 (4 hrs) without S) (4 hrs)
0 0.0 0.0
1'250 0.0 0.0
2'500 0.0 0.0
5'000 0.0 0.5
MMC 0.4 0.0 NA
CP 7.5 NA 0.0

Table 8: Mean Frequency of Polyploid Cells - Experiment 2:

Dose [µg/ml] with S9 (4 hrs) without S) (4 hrs)
0 0.5 0.0
1'250 0.0 0.0
2'500 0.0 0.0
5'000 0.0 0.5
MMC 0.4 0.0 NA
CP 7.5 NA 0.0

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

The test material was considered to be non-clastogenic to human lymphocytes under the conditions tested.