Reaction mass of 1-O-α-D-glucopyranosyl-D-mannitol and 6-O-α-D-glucopyranosyl-D-glucitol

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:

14 Nov to 19 Nov 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted in 1997

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Food and Consumer Product Safety Authority (VWA), den Haag, The Netherlands

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 1-O-α-D-glucopyranosyl-D-mannitol and 6-O-α-D-glucopyranosyl-D-glucitol

EC Number:

908-700-6

Cas Number:

64519-82-0

Molecular formula:

6-O-alpha-D-Glucopyranosyl-D-sorbitol: C12H24O11 1-O-alpha-D-Glucopyranosyl-D-mannitol: C12H24O11

IUPAC Name:

Reaction mass of 1-O-α-D-glucopyranosyl-D-mannitol and 6-O-α-D-glucopyranosyl-D-glucitol

Method

Target gene:

his operon and trp operon

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254

Test concentrations with justification for top dose:

all strains: 62, 185, 556, 1667, 5000 µg/plate

Vehicle / solvent:

- Vehicle/solvent used: milli-Q water

Controls

Untreated negative controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

benzo(a)pyrene

ethylnitrosurea

other: 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)
- Cell density at seeding: 10E8 to 10E9

DURATION
- Preincubation period: 10 -16 h
- Exposure duration: 72 h

NUMBER OF REPLICATIONS: 3 replications in one experiment

DETERMINATION OF CYTOTOXICITY
- Method: reduction (by at least 50%) in the number of revertant colonies and/or a clearing of the background lawn of bacterial growth as compared to the negative (vehicle) control

Evaluation criteria:

A test substance is considered to be positive in the bacterial gene mutation test if the mean number of revertant colonies on the test plates is concentration-related increased or if a reproducible two-fold or more increase is observed compared to that on the negative control plates.

A test substance is considered to be negative in the bacterial gene mutation test if it produces neither a dose-related increase in the mean number of revertant colonies nor a reproducible positive response at any of the test points.

Statistics:

No statistical analysis was performed.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium, other: T 1535, TA 1537, TA 98, 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 2

Key result

Species / strain:

E. coli WP2 uvr A

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

Table 3. Number of revertants counted in bacterial reverse mutation test

		TA 1535		TA 1537		TA98		TA 100		E. Coli
		-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
0 µg/plate		32	16	16	23	31	60	185	217	46	34
		26	10	6	6	37	60	163	205	46	38
		13	31	13	24	26	61	145	226	34	32
	Mean	24	19	12	18	31	60	164	216	42	35
	StDev	10	11	5	10	6	1	20	11	7	3
62 µg/plate		18	28	16	20	32	58	150	159	44	46
		17	17	17	16	32	47	198	180	35	35
		23	14	14	22	20	50	291	157	44	41
	Mean	19	20	16	19	28	52	213	165	41	41
	StDev	3	7	2	3	7	6	72	13	5	6
185 µg/plate		19	24	8	22	29	44	143	169	42	44
		30	20	14	17	38	42	136	162	37	32
		20	23	13	13	38	40	269	161	35	30
	Mean	23	22	12	17	35	42	183	164	38	35
	StDev	6	2	3	5	5	2	75	4	4	8
556 µg/plate		16	16	8	19	35	43	***	133	30	32
		18	18	16	19	29	***	141	130	38	41
		14	13	19	23	22	56	174	121	25	40
	Mean	16	16	22	20	29	50	158	128	31	38
	StDev	2	3	15	2	7	9	23	6	7	5
1667 µg/plate		13	24	13	8	24	40	129	144	34	22
		10	13	5	20	23	55	143	150	34	25
		7	6	8	***	34	54	154	137	43	52
	Mean	10	14	9	14	27	50	142	144	37	33
	StDev	3	9	4	8	6	8	13	7	5	17
5000 µg/plate		12	16	7	14	38	46	127	136	32	38
		29	20	11	16	29	50	136	117	32	25
		19	14	8	28	23	47	129	106	29	35
	Mean	20	17	9	19	30	48	131	120	31	33
	StDev	9	3	2	8	8	2	5	15	2	7
Pos. control		659	558	4339	342	956	1673	674	1494	323	1784
		659	609	3154	264	688	1441	709	1565	307	1613
		620	519	3881	383	1261	1238	641	1719	358	1533
	Mean	646	562	3791	330	968	1451	675	1593	329	1643
	StDev	23	45	598	60	287	218	34	115	26	128

*** plate not counted due to contamination

Applicant's summary and conclusion

Conclusions:

Based on the results of the conducted study the test item did not exhibit mutagenic properties in bacterial cells.