Methyl α-D-glucoside

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:

2017-05-05 to 2017-06-29

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted July 21, 1997

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

30 May, 2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Methyl α-D-glucoside

EC Number:

202-571-3

EC Name:

Methyl α-D-glucoside

Cas Number:

97-30-3

Molecular formula:

C7H14O6

IUPAC Name:

methyl α-D-glucoside

Test material form:

solid

Method

Target gene:

his-

Species / strain

Species / strain / cell type:

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

Additional strain / cell type characteristics:

DNA polymerase A deficient

Metabolic activation:

with and without

Metabolic activation system:

S9 liver Mix

Test concentrations with justification for top dose:

31.6, 100, 316, 1000, 3160 and 5000 µg due to absence of cytotoxicity in a preliminary test.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: purified water

Controls

Untreated negative controls:

yes

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

other: 2-Aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: Six concentrations ranging from 31.6 to 5000 µg alpha methyl glucoside/plate were employed in the plate incorporation test and in the preincubation test, each carried out without and with metabolic activation.
- Cell density at seeding (if applicable): E008-E009 cells/mL

DURATION
- Preincubation period: 20 min
- Exposure duration: 48h

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: other: Cytotoxicity is evidenced by a reduction in the number of spontaneous revertants by at least 50%, a clearing or diminution of the background lawn or by the degree of survival of the treated cultures.

Rationale for test conditions:

as recommended in the guideline

Evaluation criteria:

The results of the negative and positive control cultures should be within the range of the historical data generated by the testing laboratory. The range of spontaneous reversion frequencies per plate is based on Kirkland (1990):
TA98: 20 - 60
TA100: 100 - 200
TA102: 240 - 320
TA1535: 10 - 35
TA1537: 3 - 20

Where concurrent negative or positive control data fall outside the range, they may be acceptable and considered for the inclusion into the historical control distribution as long as these data are not extreme outliers.

A test item is considered to show a positive response if
- the number of revertants is significantly increased (p ≤ 0.05, U-test according to MANN and WHITNEY) compared to the solvent control to at least 2-fold of the solvent control for TA98, TA100, TA1535 and TA1537 and 1.5-fold of the solvent control for TA102 in both independent experiments.
Or
- a concentration-related increase over the range tested in the number of the revertants per plate is observed. The Spearman's rank correlation coefficient may be applied.
- positive results have to be reproducible and the histidine independence of the revertants has to be confirmed by streaking random samples on histidine-free agar plates.
Biological relevance of the results should be considered first.
A test item for which the results do not meet the above mentioned criteria is considered as non-mutagenic in the AMES test.

Results and discussion

Test results

Test results 1

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

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

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

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

Test results 5

Species / strain:

S. typhimurium TA 102

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

Any other information on results incl. tables

Historical control data:

positive controls

Strain S9-Mix	TA98		TA100		TA102		TA1535		TA1537
	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
	2-nitro-fluorene	Benzo[a] pyrene	sodium azide	2-amino-anthracene	Mito-mycin C	Benzo[a] pyrene	sodium azide	2-amino-anthracene	9-amino-acidine	Benzo[a] pyrene
Mean	151.2	150.4	952.9	948.8	1029.7	1024.3	135.6	135.4	76.7	77.6
SD	27.9	28.8	99.7	103.7	97.1	97.1	28.8	28.4	26.5	26.4
Min	91	96	677	703	781	781	51	49	28	31
Max	293	291	1213	1195	1637	1366	266	270	185	184

negative controls:

Strain S9-Mix	TA98		TA100		TA102		TA1535		TA1537
	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
Mean	30.3	32.1	145.1	145.2	277.3	279.0	19.7	19.9	6.7	6.7
SD	5.6	5.9	18.4	19.2	16.5	17.2	4.4	4.6	1.7	1.8
Min	20	20	107	101	245	203	10	10	2	3
Max	49	49	195	198	323	324	34	36	10	10

Applicant's summary and conclusion

Conclusions:

In the present test conducted according to OECD guideline 471, the Salmonella typhimurium strains TA 98, 100, 102, 1535, 1537 were incubated with the following concentrations of alpha methyl glucoside: 31.6, 100, 316, 1000, 3160 and 5000 µg /plate. No increases in revertant colonies were observed no cytotoxicity occurred. The test item is therefore considered non mutagenic under the present conditions.

Executive summary:

In a reverse gene mutation assay in bacteria (OECD guideline 471), strains TA 98, TA 100, TA 102, TA 1535 and TA 1537of S. typhimurium were exposed to alpha methyl glucoside (100% a.i.) in highly purified water at concentrations of (31.6, 100, 316, 1000, 3160 and 5000 µg/plate in the presence and absence of mammalian metabolic activation using the plate incorporation method and the preincubation method. 

Alpha methyl glucoside was tested up to limit concentration (5000 µg/plate). There was no increase in the number of revertants regardless of strain.  The positive controls induced the appropriate responses in the corresponding strains.  There was no evidence of induced mutant colonies over background.

 

This study is classified asacceptable.  This study satisfies the requirement for Test Guideline OECD 471 for in vitro mutagenicity (bacterial reverse gene mutation) data.