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Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information
Salmonella/microsome test (Ames test): positive with strains TA 98 and TA 100 (+ S9 mix) and negative with strains TA 102, TA 1535 and TA 1537 (+/- S9 mix)
Link to relevant study records
Reference
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:
Oct to Nov 2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
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:
EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay
Target gene:
Histidine gene locus
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254 induced male rat liver S9 mix
Test concentrations with justification for top dose:
0, 50, 158, 500, 1581, 5000 µg/plate (first test, +/-S9 mix, all strains)
0, 156, 312, 624, 1248, 2496, 4992 µg/plate (repeat test, +S9 mix, TA 98 and TA 100 only)







Vehicle / solvent:
DMSO
Untreated negative controls:
yes
Negative solvent / vehicle controls:
no
Remarks:
No solvent control was used since sufficient evidence was available in the literature and from testing laboratory experience, indicating that the solvents used had no influence on the spontaneous mutant counts of the used strains.
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: sodium azide (only TA 1535), nitrofurantoin (only TA 100), 4-nitro-1,2-phenylene diamine (TA 1537 and TA 98), mitomycin C (only TA 102), 2-aminoanthracene (all strains).
Remarks:
The positive controls sodium azide, nitrofurantoin, 4-nitro-1,2-phenylene diamine and mitomycin C were only used without S9 mix; the positive control 2-aminoanthracene was only used with S9 mix.
Details on test system and experimental conditions:
METHOD: each concentration including the controls was tested in triplicate.
Evaluation criteria:
A reproducible and dose-related increase in mutant counts of at least one strain is considered to be a positive result. For TA 1535, TA 100 and TA 98 this increase should be about twice that of negative controls, whereas for TA 1537 at least a threefold increase should be reached. For TA 102 an increase of about 100 mutants should be reached. Otherwise, the result is evaluated as negative. However, these criteria may be overruled by good scientific judgment. In case of questionable results, investigations should continue, possibly with modifications, until a final evaluation is possible.
Statistics:
not specified
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
not examined
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
not examined
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
not examined
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: strain/cell type: TA 98, TA 100
Remarks:
Migrated from field 'Test system'.

Table 1: Summary of results from the Salmonella mutagenicity assay (first test) with Picolinamid-Phenylether (mean values of revertants per plate)

 Dose (µg per plate)

Without metabolic activation

 

TA 1535

 TA 100

 TA 1537

 TA 98

 TA 102

0

8

135

8

38

267

50

10

135

7

36

256

158

9

158

6

29

262

500

9

153

5

39

245

1581

9

158

5

37

218

5000

11

145

6

39

192

 Positive control

758*

317*

88*

150*

572* 

 Dose (µg per plate )

With metabolic activation (liver S9 mix)

 

TA 1535

 TA 100

 TA 1537

 TA 98

TA 102

0

11

178

11

54

269

50

12

177

8

55

249

158

11

211

9

70

305

500

11

233*

8

76

296

1581

8

243*

10

120*

276

5000

10

294*

14

183*

244

 Positive control

131*

1661*

281*

1411*

690*

 

* = mutagenic effect

 

Table 2: Summary of results from the Salmonella mutagenicity assay (repeat test with strains TA 98 and TA 100 and metabolic activation) with Picolinamid-Phenylether (mean values of revertants per plate)

 

Dose (µg per plate)

With metabolic activation (liver S9 mix)

 

TA 1535

 TA 100

 TA 1537

 TA 98

 TA 102

0

-

153

-

41

-

156

-

201

-

60

-

312

-

190 

-

65

-

624

-

221*

-

87*

-

1248

-

241*

-

101*

-

2496

-

240*

-

140*

-

4992

-

260*

-

203*

-

 Positive control

-

1592* 

1305* 

 

* = mutagenic effect

Doses up to and including 5000 µg per plate did not cause any bacteriotoxic effects. Total bacteria counts remained unchanged and no inhibition of growth was observed.

Evidence of mutagenic activity of Picolinamid-Phenylether was seen. On Salmonella typhimurium TA 100 and TA 98, a biologically relevant increase was found in the mutant count compared to the corresponding negative control. Positive response was found only with S9 mix. The lowest effective dose was 500 µg per plate for TA 100 and 624 µg per plate for TA 98. The Salmonella/microsome test thus showed Picolinamid-Phenylether to have a mutagenic effect.

The positive controls sodium azide, nitrofurantoin, 4-nitro-1,2-phenylene diamine, mitomycin C and 2-aminoanthracene had a marked mutagenic effect, as was seen by a biologically relevant increase in mutant colonies compared to the corresponding negative controls.

.
Conclusions:
Interpretation of results (migrated information):
positive with metabolic activation
Executive summary:

The mutagenic potential of Picolinamid-Phenylether was evaluated in a Salmonella/microsome test with the S. typhimurium strains TA 98, TA 100, TA 102, TA 1535 and TA 1537 in the presence and absence of S9 mix according to OECD TG 471. Doses up to and including 5000 µg per plate did not cause any bacteriotoxic effects. Total bacteria counts remained unchanged and no inhibition of growth was observed. Evidence of mutagenic activity of Picolinamid-Phenylether was seen. On Salmonella typhimurium TA 100 and TA 98, a biologically relevant increase was found in the mutant count compared to the corresponding negative control. Positive response was found only with S9 mix. The lowest effective dose was 500 µg per plate for TA 100 and 624 µg per plate for TA 98. The Salmonella/microsome test thus showed Picolinamid-Phenylether to have a mutagenic effect.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (positive)

Additional information

Additional information from genetic toxicity in vitro:

The mutagenic potential of Picolinamid-Phenylether was evaluated in a Salmonella/microsome test with the S. typhimurium strains TA 98, TA 100, TA 102, TA 1535 and TA 1537 in the presence and absence of S9 mix according to OECD TG 471 (Wirnitzer, 2004). Doses up to and including 5000 µg per plate did not cause any bacteriotoxic effects. Total bacteria counts remained unchanged and no inhibition of growth was observed. Evidence of mutagenic activity of Picolinamid-Phenylether was seen. On Salmonella typhimurium TA 100 and TA 98, a biologically relevant increase was found in the mutant count compared to the corresponding negative control. Positive response was found only with S9 mix. The lowest effective dose was 500 µg per plate for TA 100 and 624 µg per plate for TA 98. The Salmonella/microsome test thus showed Picolinamid-Phenylether to have a mutagenic effect.


Justification for selection of genetic toxicity endpoint
Only one study available

Justification for classification or non-classification

Based on the study results a classification according to Directive 67/548/EEC or Regulation (EC) No. 1272/2008 (CLP) is not warranted.