D-glucopyranose, oligomers, xylityl glycosides and 1,4 anhydro D-xylitol and D-xylitol

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

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OCDE n° 471 Dir. 92/69/CEE : B13/B14

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

LCA 01006

IUPAC Name:

LCA 01006

Constituent 2

Reference substance name:

-

EC Number:

446-990-1

EC Name:

-

Molecular formula:

C5H11O5 [C6H10O5]1.45 and C5H10O4 and C5H12O5

IUPAC Name:

2-(hydroxymethyl)oxolane-3,4-diol; 5-{[2,3,4,5-tetrahydroxy-6-(xenoniooxy)hexyl]oxy}pentane-1,2,3,4-tetrol; pentane-1,2,3,4,5-pentol

Details on test material:

- Product : LCA01006
- Product supplier : SEPPIC
- Received sterile on 19th June, 2001
- LEMI Reference : DIB190601-2

Method

Target gene:

For Salmnonella typhimurium: histidine
For Escherichia coli: tryptophane

Species / strain

Species / strain / cell type:

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

Additional strain / cell type characteristics:

other: strain TA 98, TA 100 and E coli wp2 contains PKM 101

Metabolic activation:

with and without

Metabolic activation system:

S9-Mix (10 % liver S9 in standard co-factors)

Test concentrations with justification for top dose:

Concentration range in the main test (with metabolic activation): 50 ... 5000 µg/plate
Concentration range in the main test (without metabolic activation): 50 ... 5000 µg/plate

Vehicle / solvent:

distilled water

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: cis-diammineplatinium (II) dichloride

Remarks:

for E. coli strain

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

for TA 100 strain, without S9 activation

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

for TA 98 strain, without S9 activation

Controls 4

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

for TA 1537 strain, without S9 activation

Controls 5

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: 2-anthramine

Remarks:

for all S. typimurium strains, with S9 activation

Controls 6

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: B-propiolactone

Remarks:

E. coli strain without S9

Details on test system and experimental conditions:

Assay without metabolic activation
For each of the strains, 100 μL of the bacterial suspension (1-2 x 109 bacteria/mL) and 100 μL of the test
substance are mixed with 2.0 mL of overlay agar and poured over the surface of a minimal agar plate (90
mm in diameter) (n = 3). The overlay agar is allowed to solidify before incubation.
Plates are incubated at 37°C over a 48 hour period. The number of revertant colonies per plate is counted.
Moreover the following controls are carried out :
- negative controls
. absolute negative control (spontaneous reversion rate),
. positive control solvent.
- test substance solvent if necessary.
- positive control

Assay with metabolic activation
Two tests can be performed using
· either a standard plate incorporation method where the protocol is similar to that described
above, except that just before pouring the mixture onto the plates, 500 μL of S-9 mix fraction is
quickly mixed,
· or the pre-incubation assay where the test substance is preincubated with the test strain, and
500 μL of S-9 mix fraction for 1 hour at 37°C prior to mixing with the overlay agar and pouring
onto the surface of the minimal agar plate. Tubes should be aerated during pre-incubation by
using a shaker. This method is known to increase the detection sensitivity of a number of
promutagens like alkaloïds, aliphatic N-Nitroso compounds (OECD n° 471* * * ).

Assay are conduced in triplicate

Evaluation criteria:

The result of the test is considered as negative if the revertant number is below three fold the
number of spontaneous reversions, for TA 1535 and TA 1537 strains, and below two fold the number of
spontaneous reversions for TA 98, TA 100 and Escherichia coli WP2 strains with and without metabolic
activation.

The result of the test is considered positive if a concentration –related in crease is obtained in one,
or several of the 5strains, with and/or without metabolic activation; a mutagenic effect is taken into
account for a given concentration of the test substance if the number of revertant colonies is at least two
fold that of spontaneous revertant colonies number for TA 98, TA 100 and Escherichia coli (WP2), and
three fold for TA 1535 and TA 1537.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with

Genotoxicity:

not specified

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

(up to 5000 µg/plate)

Vehicle controls validity:

valid

Test results 2

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: other: preliminary test

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation

There is no significant difference between the number of spontaneous reversions, the number of
reversions obtained in the positive controls (with and without metabolic activation), and the mean of
corresponding experimental historic values obtained in the laboratory.
There is no evidence of any increase in the number of revertant colonies in the presence of the test
substance (5 000 μg - 1 500 μg - 500 μg - 150 μg - 50 μg) with and without metabolic activation.
Results were confirmed in a second independent experiment.

Executive summary:

Analysis : Reverse mutation assay.

Test substance : LCA01006 LEMI code : DIB190601-2

Test procedure : Reverse mutation assay on “Salmonella typhimurium his-“ and “Escherichia coli” WP2 (pKM 101) strains according to the OECD guideline n° 471 (LEMI operating procedure MB08/45).

Test result : The product did not induce reverse mutation, Salmonella typhimurium and Escherichia coli WP2 (pKM 101) strains according to the OECD guideline n° 471 and according to the test method B13/B14 of the E.C. Directive n° 2000/32.