[No public or meaningful name is available]

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:

12 January 1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

OECD 471

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

Test in accordance with OECD guidelines 471, as explicitely mentioned in test report conclusion page 18 of enclosed study report

GLP compliance:

yes (incl. QA statement)

Remarks:

See quality assurance statement in report page 4

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Chondroitin disaccharide deltadi-4S sodium salt

Cas Number:

136144-56-4

Molecular formula:

C14H19NO14S•2Na

IUPAC Name:

Chondroitin disaccharide deltadi-4S sodium salt

Constituent 2

Reference substance name:

α-deltaUA-[1→3]-GalNAc-6S = α-[4-deoxy-L-threo-hex-4-enopyranosyluronic acid] sodium salt-[1→3]-N-acetyl-D-galactosamine-6-sulphate

Cas Number:

136132-72-4

Molecular formula:

C14H19NO14S•2Na

IUPAC Name:

α-deltaUA-[1→3]-GalNAc-6S = α-[4-deoxy-L-threo-hex-4-enopyranosyluronic acid] sodium salt-[1→3]-N-acetyl-D-galactosamine-6-sulphate

Constituent 3

Reference substance name:

unknown metal constituents

IUPAC Name:

unknown metal constituents

Constituent 4

Reference substance name:

Chloride

Cas Number:

16887-00-6

Molecular formula:

Cl

IUPAC Name:

Chloride

Constituent 5

Reference substance name:

Ethanol

EC Number:

200-578-6

EC Name:

Ethanol

Cas Number:

64-17-5

Molecular formula:

C2H6O

IUPAC Name:

Ethanol

Constituent 6

Reference substance name:

unknown constituent

Molecular formula:

unknown

IUPAC Name:

unknown constituent

Constituent 7

Reference substance name:

Glucuronic acid

EC Number:

229-486-4

EC Name:

Glucuronic acid

Cas Number:

6556-12-3

Molecular formula:

C6H10O7

IUPAC Name:

(2S,3S,4S,5R,6R)-3,4,5,6-Tetrahydroxyoxane-2-carboxylic acid / (2S,3S,4S,5R,6R)-3,4,5,6- Tetrahydroxyoxane-2-carboxylic acid

Constituent 8

Reference substance name:

(2R,3S,5R)-3-amino-6-(hydroxyméthyl)oxane-2,4,5-triol

Cas Number:

7535-00-4

Molecular formula:

C6H13NO5

IUPAC Name:

(2R,3S,5R)-3-amino-6-(hydroxyméthyl)oxane-2,4,5-triol

Constituent 9

Reference substance name:

group of unknown constituents

Molecular formula:

unknown

IUPAC Name:

group of unknown constituents

Test material form:

solid

Method

Target gene:

His locus

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

Remarks:

except TA102 (rfa/uvrB+)

Metabolic activation:

with and without

Metabolic activation system:

rat microsomal liver fraction (S9 mix)

Test concentrations with justification for top dose:

Test concentrations: 0, 50, 150, 500, 1500, 5000 µg/plate.
Justification for top dose: due to its solubility in aqueous medium, the test compound was dissolved in distilled water at a maximum concentraion of 50 mg/mL. After that, this solution was used at 100 µL/plate giving a final concentration of 5000 µL/plate (maximum dose recommended by OECD guideline).

Vehicle / solvent:

Distilled water

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

benzo(a)pyrene

mitomycin C

other: 2-anthramine(a)

Details on test system and experimental conditions:

0.1 ml of the test product + 0.1 ml of a bacterial suspension from a culture agitated overnight at 37°C added to 2 mL of top agar to which 10% of 0.5 mM biotin histidine solution, maintained in a state of superfusion at 45 °C, has been added. Content agitated and spread out in a Petri plate containing 20 mL of minimum agar. A triplicate test for each experimental point is made and the plates are then kept at 37°C for 48 hours after which the number of revertant colonies is determined for each plate.
In case of metabolic activation, same mecanism but 0.5 mL of the S9 mix is added before spreading in the plates.

Rationale for test conditions:

in accordance with OECD guideline 471

Evaluation criteria:

Developpement of prototrophic mutant colonies which are then counted.

Statistics:

Results may be analysed by means of Dunnett's method allowing the comparison of several treatment means to a solvent control.

Results and discussion

Test results

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 specified

Positive controls validity:

valid

Additional information on results:

The validity criteria for the test were fulfilled.

Remarks on result:

other: no mutagenic activity

Applicant's summary and conclusion

Conclusions:

Chondroitine sulfate sodium salt was non mutagenic (nor cytotoxic) on the 5 Salmonella typhimurium strains tested. The results do not allow to classify the substance.

Executive summary:

Non mutagenic in an OECD 471 Ames test in vitro, up to 5000 ug/plate on the 5 Salmonella typhimurium strains tested.