2-amino-2-methylpropane-1,3-diol

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:

29 Sep - 09 Oct 1988

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Basic data given: TA102 or E. coli WP2 are not included in the testing. According to OECD 471, at least five strains of bacteria should be included in the testing. In this Ames test, only four S. typhimurium strains (TA1535, TA1537, TA98, TA100) were used. It is known that these strains have not the potential to detect certain types of mutagens, such as cross-linking agents or oxidising mutagens. In order to detect such mutagens it is required to include S. typhimurium TA102 or to add a DNA repair-proficient strain of E.coli (e.g. E.coli WP2 uvrA or E.coli WP2 uvrA (pKM101)).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1988

Report date:

1988

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

- only 4 strains tested

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

yes

Remarks:

- only 4 strains tested

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-amino-2-methylpropane-1,3-diol

EC Number:

204-100-7

EC Name:

2-amino-2-methylpropane-1,3-diol

Cas Number:

115-69-5

Molecular formula:

C4H11NO2

IUPAC Name:

2-amino-2-methylpropane-1,3-diol

Test material form:

solid: crystalline

Details on test material:

- Analytical purity: no data
- Batch No.: 14840
- Storage condition of test material: At room temperature in the dark.

Method

Target gene:

his operon

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Details on mammalian cell type (if applicable):

Strains are regularly checked for their histidine-requirement, crystal violet sensitivity, UV-sensitivity, ampicillin resistance (TA98 and TA100) and the frequency of spontaneous revertants.

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

rat livers from adult males (Wistar or Sprague Dawley) intraperitoneally injected with a solution (20% w/v) of Aroclor 1254 (500 mg/kg bw) in corn oil

Test concentrations with justification for top dose:

with S9-mix: 100, 300, 1000, 3333, 5000
without S9-mix: 100, 300, 1000, 3333, 5000

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: milli-Q water (Millipore corp., Bedford, Mass., USA)

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

milli-Q water

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: without S9-mix: sodium azide (TA1535, 1 µg in saline), 9-aminoacridine (TA1537, 60 µg in saline, daunomycine (TA98, 2 µg in saline), methylmethanesulfonate (TA100, 650 µg in DMSO)

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

milli-Q water

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: with S9-mix: 2-aminoanthracene (TA1535, TA1537: 0.5 µg in DMSO; TA98, TA100: 5 µg in DMSO)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 2 experiments with and without S9-mix, each experiment with 3 replicate plates

DETERMINATION OF CYTOTOXICITY
- Method: viability determination

Evaluation criteria:

When the number of reverse mutation colonies increased by almost twice the solvent control or more, and reproducibilityor dependence on the dose of the test substance was observed, the result was considered positive.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES:
A preliminary toxicity determination of the test substance in TA100 was done. The percentage survival of the TA100 culture was determined by comparing the number of colonies on the solvent control plate with those on the plates containing the test substance. Based on these data, the AMPD was tested in the main test up to a concentration of 5000 µg/plate in the absence and presence of S9-mix

COMPARISON WITH HISTORICAL CONTROL DATA:
The negative and strain -specific positive control values fell within the laboratory background historical ranges indicating that the test conditions were optimal and that the metabolic activation system functioned properly.

Any other information on results incl. tables

Table 1: Results of the main test

1st experiment	number of revertants/plate: mean value of vehicle/solvent control ± SD	number of revertants/plate: mean value of positive control ± SD	max. number of revertants/plate: mean value of test material ± SD [concentration in experiment (µg/plate) resulting in max. number of revertants/plate]
without S9-mix
TA 98	17 ± 1	1621 ± 85	25 ± 1 [100]
TA 100	94 ± 11	673 ± 51	95 ± 16 [5000]
TA 1535	12 ± 3	195 ± 20	10 ± 3 [100]
TA 1537	12 ± 3	463 ± 35	9 ± 3 [100]
with S9-mix
TA 98	22 ± 2	440 ± 26	27 ± 5 [3330]
TA 100	85 ± 3	725 ± 83	95 ± 6 [100]
TA 1535	11 ± 3	358 ± 17	12 ± 4 [100]
TA 1537	7 ± 1	931 ± 59	10 ± 4 [3330]
2nd experiment	number of revertants/plate: mean value of vehicle/solvent control ± SD	number of revertants/plate: mean value of positive control ± SD	max. number of revertants/plate: mean value of test material ± SD [concentration in experiment (µg/plate) resulting in max. number of revertants/plate]
without S9-mix
TA 98	30 ± 3	1033 ± 83	46 ± 4 [1000]
TA 100	127 ± 16	883 ± 15	152 ± 17 [3330]
TA 1535	20 ± 2	210 ± 45	18 ± 8 [1000]
TA 1537	8 ± 2	273 ± 55	8 ± 3 [5000]
with S9-mix
TA 98	36 ± 5	904 ± 5	42 ± 4 [333]
TA 100	121 ± 4	1059 ± 73	139 ± 14 [100]
TA 1535	15 ± 5	203 ± 22	14 ± 3 [333]
TA 1537	6 ± 1	748 ± 23	9 ± 4 [333]

 

 

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative