D-Glucopyranose, oligomeric, C10-16(even numbered) alkyl glycosides

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in mammalian cells

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

1990-12-20 to 1991-03-27

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP study with acceptable restriction.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

Deviations:

yes

Remarks:

Colony sizing procedures were not described, and colony size was not reported for the negative and positive controls. The purity and the stability of the test substance under assay conditions were not provided.

GLP compliance:

yes

Type of assay:

mammalian cell gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

D-Glucopyranose, oligomers, decyl octyl glycosides

EC Number:

500-220-1

EC Name:

D-Glucopyranose, oligomers, decyl octyl glycosides

Cas Number:

68515-73-1

IUPAC Name:

68515-73-1

Details on test material:

- Name of test material (as cited in study report): MV#2725-153 (Alkylpolyglycoside)
- Physical state: light, yellow liquid (sponsor); clear, straw-colored liquid (testing laboratory)
- Analytical purity: No data
- Lot/batch No.: No data
- Storage condition of test material: room temperatur in the dark

Method

Target gene:

thymidine kinase locus

Species / strain

Species / strain / cell type:

mouse lymphoma L5178Y cells

Details on mammalian cell type (if applicable):

- Type and identity of media: Fischer's Medium for Leukemic Cells of Mice with 0.1 % Pluronics (treatment medium); supplemented with 10 % horse serum and 4 mM L-glutamine (expression period medium). Cloning medium was treatment medium with 20 % horse serum and 0.25 % agar.
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: N/A
- Periodically checked for karyotype stability: N/A
- Periodically "cleansed" against high spontaneous background: yes

Additional strain / cell type characteristics:

other: heterozygous at the thymidine kinase locus (TK +/-); clone 3.7.2C

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1242 and Aroclor 1254 (2:1 mixture) induced rat liver S9

Test concentrations with justification for top dose:

Mutation assay:
Trial 1: w/ metabolic activation: 13-1004 ug/mL - data only presented for concentrations up to 179 ug/mL due to higher doses prohibiting cloning
w/o metabolic activation: 7.5-565 ug/mL - data only presented for concentrations up to 101 ug/mL due to higher doses prohibiting cloning
Trial 2: w/ metabolic activation: 103-260 ug/mL - data only presented for concentrations up to 234 ug/mL due to higher doses prohibiting cloning

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: sterile, distilled water
- Justification for choice of solvent/vehicle: solubility

Controls

Controls 1

Untreated negative controls:

no

Remarks:

N/A

Negative solvent / vehicle controls:

yes

Remarks:

sterile distilled water

True negative controls:

no

Remarks:

N/A

Positive controls:

yes

Remarks:

N/A

Positive control substance:

ethylmethanesulphonate

Remarks:

without metabolic activation

Migrated to IUCLID6: 0.5 and 0.25 ul/ml

Controls 2

Untreated negative controls:

no

Remarks:

N/A

Negative solvent / vehicle controls:

yes

Remarks:

sterile, distilled water

True negative controls:

no

Remarks:

N/A

Positive controls:

yes

Remarks:

N/A

Positive control substance:

7,12-dimethylbenzanthracene

Remarks:

with metabolic activation

Migrated to IUCLID6: 5 and 2.5 ug/ml

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 4 hours
- Expression time (cells in growth medium): 2 days
- Selection time (if incubation with a selection agent): 10-12 days
- Fixation time (start of exposure up to fixation or harvest of cells): Cells were harvested after the selection time.

SELECTION AGENT (mutation assays): trifluorothymidine

NUMBER OF REPLICATIONS: 1 tube was treated per test substance group and positive control group; 2 tubes were treated for the solvent control group. 3 plates with selection agent and 3 plates without selection agent were plated per treatment tube.

NUMBER OF CELLS EVALUATED: the mutant frequency was expressed as the number of mutants per 1.0 E6 clonable cells.

DETERMINATION OF CYTOTOXICITY
- Method: % Total Growth= (% suspension growth x % cloning growth) x 100

OTHER: for expression of the mutant phenotype, the cultures were counted and adjusted to 3.0 E5 cells/ml (if the cell population exceeded 3.0 E5 cells/ml) at approximately 24 and 48 hours after treatment in 20 and 10 ml total volume, respectively.

Evaluation criteria:

The following criteria were used as guidelines in judging the significance of the activity of the test substance in the system. In evaluating the results, it was considered that increases in mutant frequencies, which occurred only at highly toxic concentrations, may have been due to epigenetic events. However, it was impossible to formulate criteria which could apply to all types of data which may have been generated and therefore the scientist's evaluation was the final endpoint.
1. Positive-if there was a positive dose response and one or more of the three highest doses in the 0% or greater Total Growth range exhibited a mutant frequency which was two-fold greater than the background level. All data including that from cultures with less than 10% Total Growth would be used to establish the dose response relationship.
2. Equivocal-if there was no dose response but any one or more of the three highest doses with 10% or greater Total Growth exhibited a two-fold increase in mutant frequency over background, or if there was a dose response but no culture exhibited a two-fold increase in mutant frequency over background.
3. Negative- if there was no dose response in cultures with 10 % or greater Total Growth and none of these test cultures exhibited a two-fold or greater increase in mutant frequency over background.

Results and discussion

Test results

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

N/A

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

see remarks on results

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES:
The optimal dose levels for the mutagenicity assays were selected following a preliminary toxicity test based on cell population growth relative to the solvent controls. L5178Y cells were exposed to solvent alone and seven concentrations of the test substance ranging from 5013 to 0.01 ug/ml for 4 hours in the absence and presence metabolic activation. Each tube was gassed with 5 +/- 1 % CO2 in air and placed on a Bellco roller drum apparatus rotating at approximately 25 rpm. The final solvent concentration in the culture medium was 1 % by volume. The test solutions were prepared under amber light s and kept in darkness during the entire exposure period. After approximately 4 hours, the test substance in solution was removed by centrifuging the cells at 1000 rpm for 10 minutes and decanting the supernatant. The cells were washed twice in 10 ml of medium (containing serum), resuspended in 20 ml of medium (containing serum), gassed with 5 +/- 1 % CO2 in air, and replaced on the roller drum apparatus. Cell population density was determined 24 and 48 hours after the initial exposure to the test substance by removing a sample from each treatment tube, diluting in 0.1 % trypsin, incubating at 37 +/- 1 degrees C for 10 minutes, and counting the samples with an electronic cell counter. The cultures were adjusted to 2.0 E5 cells/ml (if the cell population exceeded 3.0 E5 cells/ml) after 24 hours, only.
The toxicity test indicted 100 % toxicity at 1003 ug/ml for both the non-activated cultures and for the activated cultures.

ADDITIONAL INFORMATION ON CYTOTOXICITY: See below

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'. Remarks: N/A

Any other information on results incl. tables

After a two day expression period ten doses of the non-activated and the activated cultures were selected for cloning. The non-activated cultures were cloned with 101, 75, 57, 42, 32, 24, 18, 13, 10 and 7.5 ug/ml which produced a range in suspension growth of 39% to 100%. The activated cultures were cloned with 179, 134, 101, 75, 57, 42, 32, 24, 18 and 13 µg/mL. these concentrations produced a range in suspension growth of 61% to 106%.

None of the non-activated cultures that were cloned exhibited a mutant frequency which was at least two times the mean mutant frequency of the solvent controls. The total growths of these cultures ranged from 39% to 104%. A dose-dependent response was not noted in the treated cultures.

None of the activated cultures that were cloned exhibited a mutant frequency which was at least two times the mean mutant frequency of the solvent control. The total growths of these cultures ranged from 68% to 117%. A dose-dependent response was note noted in the treated cultures.

In the second trial the activated cultures cloned were treated with 234, 229, 225, 221, 210, 199, 189, 180, 171 and 161 µg/mL.. These concentrations produced a range in suspension growth of 12% to 71%. None of these cultures exhibited a mutant frequency which was at least two times the mean mutant frequency of the solvent control. The total growths of these cultures ranged from 11% to 73%. A significant dose-dependent response was not noted in the treated cultures.

The solvent and positive controls fulfilled the requirements for a valid test.

 

Applicant's summary and conclusion

Conclusions:

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

D-Glucopyranose, oligomeric, decyl octyl glycosides was evaluated in the L5178Y TK+/-Mouse Lymphoma Mutagenesis Assay in the absence and presence of Aroclor-induced rat liver S9. Under the conditions of the assay, the test substance was found to be negative for mutagenic activity.

Executive summary:

D-Glucopyranose, oligomeric, decyl octyl glycosides (alkylpolyglycoside, 20% in ethanol) was evaluated in the L5178Y TK +/-Mouse Lymphoma Mutagenesis Assay in the absence and presence of Aroclor-induced rat liver S9. The assay was performed in two phases. The first phase, the preliminary toxicity assay, was used to establish the dose range for the mutagenesis assay. The second phase, the mutagenicity assay, was used to evaluate the mutagenic potential of the test substance. In the preliminary toxicity assay, the test substance was evaluated for its effects on the growth of L5178Y TK +/- cells at concentrations ranging from 5013 to 0.01 µg/mL in the presence and absence of S9 metabolic activation. The preliminary toxicity test indicated 100% toxicity at 1003 µg/mL for both the non-activated and the S9 activated cultures. Based on the results of the toxicity test, dose levels ranging from 565 to 7.5 µg/mL were chosen to be tested without S9 metabolic activation and dose levels ranging from 1004 to 13 µg/mL were tested with S9 metabolic activation. The S9 activated portion was repeated over a range from 260 to 103 µg/mL to achieve lower Total Growths. After a 4-hour treatment period and a two-day expression period, ten doses of the non-activated and the S9 activated cultures were selected for cloning. The non-activated cultures were cloned with 101, 75, 57, 42, 32, 24, 18, 13, 10, and 7.5 µg/mL, which produced a range in Suspension Growth of 39% to 100%. The S9 activated cultures were cloned with 179, 134, 101, 75, 57, 42, 32, 24, 18, and 13 µg/mL. These concentrations produced a range in Suspension Growth of 61% to 106%. 

None of the non-activated cultures that were cloned exhibited a mutant frequency which was at least two times the mean mutant frequency of the solvent controls. The Total Growths of these cultures ranged from 39% to 104%. A dose-dependent response was not noted in the treated cultures.

None of the S9 activated cultures that were cloned exhibited a mutant frequency which was at least two times the mean mutant frequency of the solvent controls. The Total Growths of these cultures ranged from 68% to 117%. A dose-dependent response was not noted in the treated cultures.

In the second trial the S9 activated cultures were cloned with 234, 229, 225, 221, 210, 199, 189, 180, 171, and 161 µg/mL. These concentrations produced a range in Suspension Growth of 12% to 71%. None of these cultures exhibited a mutant frequency which was at least two times the mean mutant frequency of the solvent controls. The Total Growths of these cultures ranged from 11% to 73%. A significant dose-dependent response was not noted in the treated cultures.   

The solvent and positive controls fulfilled the requirements for a valid test.

Under the conditions of the assay described in the study report, the test substance was found to be negative in both the absence and presence of exogenous metabolic activation.