1-chloro-N,N-diethyl-1,1-diphenyl-1-(phenylmethyl)phosphoramine

Genetic toxicity: in vitro

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:

July 1983

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: 5 bacterial strains tested, but the study does not include Salmonella typhimurium TA102 or Escherichia coli WP2 uvrA (pKM101))

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

1983

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

GM102E

IUPAC Name:

GM102E

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

white powder

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium, other: TA100, for the preliminary test.

Species / strain 2

Species / strain / cell type:

S. typhimurium, other: TA 1535, TA 1537, TA 1538, TA 98 and TA 100.

Metabolic activation:

with and without

Metabolic activation system:

Rat S9

Test concentrations with justification for top dose:

Concentration range in the main tests (with metabolic activation): 10, 33, 100, 333, 1000, 3300 µg/plate
Concentration range in the main tests (without metabolic activation): 10, 33, 100, 333, 1000, 3300 µg/plate

Concentrations of two main tests were determined after a range finding study performed on the following conentrations: 33 µg, 100 µg, 333 µg, 1 mg, 3.3 mg and 10 mg per plate.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: soluble

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

other: 2-Aminoanthracene

Remarks:

With S9: 2-Aminoanthracene ; Without S9: the other positive control substances

Details on test system and experimental conditions:

RANGE FINDING TEST
One plate per exposure level.
Application in Agar (plate incorporation).

MAIN TESTS
Two main tests performed.
Triplicates.
Application in Agar (plate incorporation).

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium, other: TA 100 for the preliminary test.

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium, other: TA 100 for the preliminary test.

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium, other: TA 1535, TA 1537, TA 1538, TA 98 and TA 100 for the main test #1

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

thin background lawn at 3330 µg/plate for TA1535 and TA1538

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium, other: TA 1535, TA 1537, TA 1538, TA 98 and TA 100 for the main test #1

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

>= 1000 µg/plate for TA1538 and TA100 ; 333 µg/plate for TA1535; 3300 µg/plate for TA1537 and TA98

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 5

Species / strain:

other: TA 1535, TA 1537, TA 1538, TA 98 and TA 100 for the main test #2

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 3300 µg/plate for TA1535, TA98 and TA1538

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 6

Species / strain:

other: TA 1535, TA 1537, TA 1538, TA 98 and TA 100 for the main test #2

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

>= 333 µg/plate for TA1538 and TA1535 ; at 3300 µg/plate for TA1537, TA100 and TA98

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

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

GM102E was not mutagenic in any of the 5 strains of S. typhimurium used in the study. The test substance was toxic to bacteria from a concentration of 333 µg per plate.

Executive summary:

In a reverse gene mutation assay in bacteria, performed according to the OECD No.471 guideline and in compliance with GLP, GM102E diluted in DMSO was tested in S. typhimurium TA 1535, TA 1537, TA 1538, TA 98 and TA 100 in the presence and the absence of mammalian metabolic activation (S9). Four known mutagens (Sodium azide; 2-Aminoanthracene; 2-nitrofluorene and 9-aminoacridine), dissolved in dimethylsulfoxide (except for sodium azide which was dissolved in sterile distilled water), were used to check the sensitivity of the test system.

A preliminary study (one plate/concentration) was performed in order to determine the appropriate concentrations for the two independent main studies (3 plates/concentration). In the preliminary study, the bacterial strain TA100 was exposed to the test substance at the following concentrations: 33 µg, 100 µg, 333 µg, 1 mg, 3.3 mg and 10 mg per plate. Cytotoxicity, assessed by the decrease in the number of revertants and/or the thinning of the bacterial lawn, was observed from a concentration of 3.3 mg/plate in the presence of S9-mix (a moderate to strong toxicity was noted at 10 mg/plate) and from 1 mg/plate in the absence of S9-mix (a strong toxicity was noted at dose-levels ≥ 3.3 mg/plate).

In the main studies GM102E was tested with and without S9 mix at 10, 33, 100, 333, 1000, 3300 µg/plate for the TA 1535, TA 1537, TA 1538, TA 98 and TA 100 strains in both experiments. The results of the two main tests showed that toxicity was observed in the presence of S9-mix in strains TA 1535 and TA 1538 at the top dose tested of 3.3 mg/plate. In the absence of S9-mix, toxicity was noted from 333 µg/plate in strain TA 1535, from 1 mg/plate in strain TA 1538 and at 3.3 mg/plate in strains 1537, TA 98 and TA100. The number of revertants for the vehicle and positive controls met the acceptance criteria. The study was therefore considered to be valid. The test item did not induce any noteworthy or biologically relevant increase in the number of revertants, in any of the other tested strains.  

Under the test conditions, GM102E did not show any mutagenic activity in the bacterial reverse mutation test using Salmonella typhimurium. This study is considered as acceptable as it satisfied the criteria of the OECD Guideline No. 471.