Allyl (3-methylbutoxy)acetate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Allyl (3-methylbutoxy)acetate

EC Number:

266-803-5

EC Name:

Allyl (3-methylbutoxy)acetate

Cas Number:

67634-00-8

Molecular formula:

C10H18O3

IUPAC Name:

allyl (3-methylbutoxy)acetate

Test material form:

liquid

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

The mammalian liver post-mitochondrial fraction, 10% Mutazyme, is a pre-mixed fraction with all co-factors including glucose-6-phosphate, NADP, MgCl2, KCl and rat liver S-9 MolToxTM S-9 were stored frozen in aliquots at -20 ºC and thawed prior to use.

Test concentrations with justification for top dose:

The test item was tested for mutation in five strains of Salmonella typhimurium of exactly 108 cells per mL (TA98, TA100, TA1535, TA1537 and TA102) and at the concentrations of 0.2, 0.4, 0.6, 0.8, 1, 2 µL/plate using triplicate plates with and without S-9 mix.

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

benzo(a)pyrene

mitomycin C

Details on test system and experimental conditions:

All the test item concentrations were administered to the test system within 4 h and 15 mins of preparation. Once set, the plates were inverted and incubated at 37 C for 48 h (15 May 2018, 1:55 p.m. to 17 May 2018, 1:55 p.m.). Plating was achieved as described in the range finder experiment.

Rationale for test conditions:

according to the guideline

Evaluation criteria:

The test item was considered to be mutagenic in this assay if:
1. The assay is valid.
2. A clear increase in mutation frequencies induced by test item compared to the
concurrent solvent controls.
Results which partially satisfy the above criteria was dealt with on a case-by-case basis.
Biological relevance was taken into account, for example consistency of response within and between concentrations and (where applicable) between experiments.

Statistics:

not specified

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

Based upon the results obtained in this study and in line with OECD Guideline for Testing of Chemicals, 471 (adopted on 21st July 1997) it is concluded that, the
given test item Allyl Amyl Glycolate (Batch No. - AAG-TEST 1) is non-mutagenic.

Executive summary:

The test item, Allyl Amyl Glycolate (Batch No. - AAG-TEST 1) was assayed for its
ability to induce mutation in histidine-requiring five strains ofSalmonella typhimurium
(TA98, TA100, TA1535, TA1537 and TA102) both in the absence and in the presence
of metabolic activation by an Aroclor 1254-induced rat liver post-mitochondrial
fraction (S-9).
An initial toxicity Range-Finder Experiment was carried out in strain TA100,
both in the absence and presence of S-9, using six different concentrations (0.0016,
0.008, 0.04, 0.2, 1 and 5 µL/plate) of the test item, positive and negative controls
(in triplicates). The test item which was completely solubilized in DMSO (Dimethyl
Sulphoxide) were administered to the test system within 2 h and 49 mins. Evidence of
toxicity with moderately reduced background lawn was observed in 1 µL/plate and
inhibition of background lawn was observed in 5 µL/plate concentration following
treatments. As per OECD 471, the test items that are cytotoxic below 5 µL/plate should
be tested up to a cytotoxic concentration. Hence 2 µL/plate was selected as the top dose
for main experiment.
The main experiment, were performed with the fiveSalmonella typhimuriumtester
strains (TA98, TA100, TA1535, TA1537 and TA102). Based on the results of the range
finding experiment, the test item was tested at concentrations of 0.2, 0.4, 0.6, 0.8, 1,
2 µL/plate along with positive and negative controls (in triplicates) both in the absence
and presence of S-9. All the test item concentrations were administered to the test
system within 4 h and 15 mins. Moderately reduced background lawn was observed in
1 µL/plate concentration and complete inhibition of background lawn was observed in
2 µL/plate concentration as the evidence of cytotoxicity.
The mean numbers of revertant colonies on negative control plates fell within historical
ranges, and an increase in the revertant colonies in the positive control chemicals
confirms the discrimination between different strains, and an active S-9 preparation.
Not more than 5% of the plates were lost through contamination or some other
unforeseen event. Hence, the assay was considered valid.
No dose-related and reproducible increase in revertant colonies were observed in all
strains following test item administration, both in the absence and presence of metabolic
activation, thereby providing evidence that the test item does not mediated mutagenic
activity.
Based upon the results obtained in this study and in line with OECD Guideline
for Testing of Chemicals, 471 (adopted on 21st July 1997) it is concluded that, the
given test item Allyl Amyl Glycolate (Batch No. - AAG-TEST 1), supplied by
MORAYA GLOBAL LIMITED, is non-mutagenic.