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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:
1998-01-15 to 1998-06-23
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1998
Report Date:
1998

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
Qualifier:
according to
Guideline:
other: UKEMS Guidelines
Deviations:
not specified
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Lot/batch No.: 6108
- clear colourless liquid
- Purity : 100%
- Expiration date of the lot/batch: march 1999
- Storage condition of test material: at room temperature in the dark

Method

Target gene:
histidine gene
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 102
Metabolic activation:
with and without
Metabolic activation system:
A mammalian liver post-mitochondrial fraction (S-9) was prepared from male Sprague Dawley rats induced with Aroclor 1254 and obtained from Molecular Toxicilogy Incorporated, USA.
Test concentrations with justification for top dose:
-Mutation experiment 1 :
without S-9 : Final concentration : (µg/plate) : 0.032 ; 0.16 ; 0.8 ; 4 ; 20
with S9 : Final concentration : (µg/plate) : 0.16 ; 0.8 ; 4 ; 20 ; 100

- Mutation experiment 2 :
without S-9 (except TA1535) : 0.01953 ; 0.078125 ; 0.3125 ; 1.25 ; 5 ; 20
All strains with S-9 and strain TA1535 without S-9 : 0.078125 ; 0.3125 ; 1.25 ; 5 ; 5 ; 20 ; 80
Vehicle / solvent:
solvent used: DMSO
Controls
Untreated negative controls:
yes
Remarks:
DMSO
Negative solvent / vehicle controls:
yes
Remarks:
DMSO for 2NF, AAC and AAN // water for NaN3 and GLU
True negative controls:
no
Positive controls:
yes
Remarks:
2-nitrofluorene (2NF), sodium azide (NaN3), 9-aminoacridine (AAC), glutaraldéhyde (GLU) and 2-aminoanthracene (AAN)
Evaluation criteria:
The test article was considered to be mutagenic if :
- the assay was valid
- a dose related and reproductible increase in the number of revertants is observed, or a signifiant* and reproductible increase in the number of revertants was induced at one or more test concentration.

* an increase in revertant numbers was considered to be signifiant if the number of revertant colonies was at least two times the mean negative control counts in strains TA98, TA100 and TA102, or three times in strains TA1535 and TA1537.

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
40 ug/plate wo/metabolic activation (-S9) and 200 ug/plate w/metabolic activation (+S9)
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'. Remarks: TA102 was also tested

Any other information on results incl. tables

Table 1 Summary of mean revertant colonies (no metabolic activation) Experiment 1

        Test type
 Dose  TA98 TA100  TA1535  TA1537  TA102 
 Vehicle only  25 +/-4  110 +/-11 13 +/-  2  15 +/-3 410 +/-  58
 0.032 ug/plate 32 +/-  3  110 +/-8 14 +/-  6  15 +/-2 462 +/-  37
 0.16  ug/plate  22 +/-5  113 +/-10  13 +/-2  12 +/-3  429 +/-9
 0.8  ug/plate  26 +/-12  106 +/-8  10 +/-2  15 +/-7  428 +/-31
 4  ug/plate  25 +/-2  104 +/-4  17 +/-5  12 +/-3  376 +/-51
20  ug/plate  21 +/-2  63 +/-5  13 +/-2  12 +/-2  176 +/-27

 T toxic, S slightly thinning

Table 2 Summary of mean revertant colonies (with metabolic activation) Experiment 1

        Test type
 Dose  TA98 TA100  TA1535  TA1537  TA102 
 Vehicle  33 +/-6 131 +/-9 23 +/-3 13 +/-3 500 +/-34
 0.16  ug/plate  31 +/-1 140 +/-15 14 +/-2  17 +/-3 519 +/-26
 0.8 ug/plate  37 +/-9 127 +/-  20 +/-4  15 +/-2  463 +/-10
 4 ug/plate  31 +/-8  130 +/-8  20 +/-9  9 +/-4  436 +/-39
20  ug/plate  31 +/-5  153 +/-7  15 +/-5  13 +/-7  496 +/-11
100  ug/plate  17 +/-4 (S)  T  16 +/-3 (S)  T  T

T toxic, S slightly thinning

Table 3 Summary of mean revertant colonies (no metabolic activation) Experiment 2

        Test type
 Dose  TA98 TA100  TA1535  TA1537  TA102 
 Vehicle  34+/-6 111 +/-12 17 +/-6 15 +/-5 482 +/-22
 0.01953 ug/plate 39 +/-3 114 +/-6 13 +/-2 500 +/-15
 0.078125 ug/plate 34 +/-1 131 +/-14 12 +/-8 15 +/-5 428 +/-20
 0.3125 ug/plate 35 +/-4 119 +/-6 14 +/-4 12 +/-3 419 +/-32
1.25 ug/plate 36 +/-1 122 +/-6 14 +/-2 13 +/-3 460 +/-14
5 ug/plate 39 +/-2  134 +/-7  11+/-1  17 +/-5  452 +/-25
 20 ug/plate  20 +/-4  83 +/-2 (M+V)  13 +/-3  11 +/-3 (S)  191 +/-28 (M+V)
 80 ug/plate      4 +/-2 (M+A)    

M manual count, V very thin background lawn, A absence of background lawn, S slight thinning

Table 4 Summar of mean revertant colonies (with metabolic activation) Experiment 2

        Test type
 Dose  TA98 TA100  TA1535  TA1537  TA102 
 Vehicle 44 +/-10 169 +/-15 18 +/-2 17 +/-5 552 +/-50
 0.078125 ug/plate 40 +/-8 170 +/-9 18 +/-7 19 +/-5 613 +/-5
 0.3125 ug/plate 44 +/-8 168 +/-16 21 +/-7 17 +/-3 555 +/-16
 1.25 ug/plate 50 +/-7 155 +/-9 20+/-3 16 +/-3 519 +/-8
5 ug/plate 44 +/-7 157 +/-14 14 +/-2 14+/-1 585 +/-22
20 ug/plate 55 +/-6  160 +/-5 22 +/-2 17 +/-5 591 +/-35
 80 ug/plate 32 +/-11 (S) 150 +/-5 (S)  16 +/-2  19 +/-3 (S) 416 +/-51 (S)
         

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative with metabolic activation
ambiguous without metabolic activation

It was concluded that 2-Propanol, 1,1'-[[3-[(3-aminopropyl)amino]propyl]imino]bis-, N-tallow alkyl derivs. did not induce mutation in five strains of Salmonella typhimurium (TA98, TA100, TA1535, TA1537 and TA102), when tested under the conditions employed for this study, which included treatments approaching, or extending into, the toxic range, both in the absence and in the presence of a rat liver metabolic activation system (S-9).
Executive summary:

The potential of 2-Propanol, 1,1'-[[3-[(3-aminopropyl)amino]propyl]imino]bis-, N-tallow alkyl derivs. inducing mutations was assayed in 5 bacterial models; TA98 (frameshift), TA100 (basepair substitution), TA102 (basepair substitution), TA1535 (basepair substitution), and TA1537 (frameshift). The test item was diluted in anhydrous DMSO immediately before added to test plates. Negative control (vehicle only) and positive controls (2 -nitrofluorene, Sodium Azide, 9 -aminoacridine, Glutaraldehyde and 2 -aminoanthracene) were included. The tests were carried out with/without metabolic activation (S9 fraction from Aroclor 1254 treated rats). Colonies were counted electronically.

In an initial rangefinder assay, the test item was added to plates at a concentration of 8, 40, 200, 1000, and 2000 ug/plate. Significant toxicity was observed at 40 ug/plate (-S9) and 200 ug (+S9). Based on these studies the upper dose limits were set at 20 ug/plate (-S9) and 100 ug/plate (+S9) for the mutagenicity assessment. Polyram SL was fully soluble at the concentrations tested.

2-Propanol, 1,1'-[[3-[(3-aminopropyl)amino]propyl]imino]bis-, N-tallow alkyl derivs. did not induce mutations in any of the five strains tested and at the dose conditions listed. The positive controls performed with satisfaction.