Glycine, N,N'-1,2-ethanediylbis-, reaction products with formaldehyde, iron chloride (FeCl3) and phenol, potassium salts

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

March-April 1987

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1987

Report date:

1987

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

In deviation from the current guideline, no E Coli WP2 strain or TA 102 strain were tested. These strains contain AT base pairs, and should be used to also detect certain oxidising mutagens, cross-linking agents and hydrazines. However chelating agents as such have no chemical reactivity other than binding metal ions, so they are not considered oxidising mutagens, cross'linking agents and are not hydrazines.

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Acetic acid, oxo-, sodium salt, reaction products with cresol and ethylenediamine, iron sodium salts

EC Number:

283-041-9

EC Name:

Acetic acid, oxo-, sodium salt, reaction products with cresol and ethylenediamine, iron sodium salts

Cas Number:

84539-53-7

Molecular formula:

non specified (UVCB substance)

IUPAC Name:

non specified (UVCB substance)

Details on test material:

Name of test compound: Bolikel Fe 100SMG
Appearance: fine red-brown powder
Receipt: 27 February 1987
Storage: room temperature protected from light
Purity: 60% (no further info)

Method

Target gene:

point mutations (mutation: growth possible in the absence of an essential amino acid)

Species / strain

Species / strain / cell type:

other: Salmonelle typhimurium TA 1535, 1537, 1538, 100 and 98

Details on mammalian cell type (if applicable):

- Type and identity of media: nutrient broth (Oxoid No. 2)
- Properly maintained: yes
- Each freshly thawed batch checked for appropriate amino acid requirement, spontaneous reversion rate and growth inhibition

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced rat liver (S9 mix)

Test concentrations with justification for top dose:

Preliminary test (using TA98): 2.5 - 5000 µg/plate
Main test I + II: 50-5000 µg/plate (5 concentrations)

Vehicle / solvent:

Distilled water

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

withouit S9 mix: TA1535 / TA100

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene TA 1535

Remarks:

with and without S9 mix

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

without S9 mix: TA1537

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

without S9 mix: TA1538 / TA98

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

with and without S9 mix: TA1537 / TA1538 / TA100 / TA98

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar

DURATION
- Preincubation period: overnight
- Exposure duration: 2 days
- Expression time (cells in growth medium): not applicable
- Selection time (if incubation with a selection agent): not applicable
- Fixation time (start of exposure up to fixation or harvest of cells): not applicable

SELECTION AGENT (mutation assays): not applicable
SPINDLE INHIBITOR (cytogentic assays): not applicable
STAIN (for cytogenetic assays): not applicable

NUMBER OF REPLICATIONS: in triplicate

NUMBER OF CELLS EVALUATED: not applicable

DETERMINATION OF CYTOTOXICITY
- Method: background lawn, revertant colonies

OTHER EXAMINATIONS: none

Evaluation criteria:

Not indicated; most likely an increase in the number of revertants

Statistics:

Not needed

Results and discussion

Test results

Species / strain:

other: TA1535, 1537, 1538, 98 and 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

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: no info
- Effects of osmolality: no info
- Evaporation from medium: not likely (dust)
- Water solubility: no problem due to high water solubility
- Precipitation: no
- Other confounding effects: no info

RANGE-FINDING/SCREENING STUDIES: no visible thinning of background lawn of non-revertant cells was observed up to 5000
µg/plate

COMPARISON WITH HISTORICAL CONTROL DATA: no info

ADDITIONAL INFORMATION ON CYTOTOXICITY: see above

Applicant's summary and conclusion

Conclusions:

No mutagenic effect was observed in the present study up to levels of 5000 µg/plate.

Executive summary:

The compound Bolikel Fe 100SMG was examined for mutagenic activity in five histidine-dependent auxotrophs of Salmonella typhimurium strains TA 98, TA 1538, TA 100, TA 1535 and TA 1537, using pour-plate assays. The procedures used complied with the OECD Guidelines for testing of chemicals (1983) and the EPA Toxic Substances Control Act Test Guidelines (1985). Each test, in each strain, was conducted on two separate occasions. The studies, which were conducted in the absence and presence of an activating system derived from rat liver (S-9 mix), employed a range of levels of Bolikel Fe 100SMG from 50 to 5000 µg per plate, selected following a preliminary toxicity test in strain TA 98, and included solvent (distilled water) controls with and without S-9 mix. No increases in reversion to prototrophy were obtained with any of the five bacterial strains at the compound levels tested, either in the presence or absence of S-9 mix. Marked increases in the number of revertant colonies were induced by the known mutagens benzo[a]pyrene, 2 -nitrofluorene, 2 -aminoanthracene, 9 -aminoacridine and sodium azide when examined under similar conditions. It was concluded that Bolikel Fe 100SMG was devoid of mutagenic activity under the conditions of the test.