2-Hydroxypropyl-β-cyclodextrine ethers

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Link to relevant study records

Reference

Reference 1

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)

Rationale for reliability incl. deficiencies:

other: Comparable to OECD Guideline Study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

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

Species / strain / cell type:

E. coli WP2 uvr A pKM 101

Metabolic activation:

with and without

Metabolic activation system:

S9-mix

Test concentrations with justification for top dose:

Positive control (µg/plate): 1.0 (All Salmonella Strains), 10 (WP2 uvrA), 100 (WP2)
Positive control (µg/plate): 1.0 (TA98, TA1538), 1.0 (TA100, TA 1535), 75 (TA1537), 1,000 (Both E.coli Strains)
Vehicle (µg/plate): 6.5, 9.7, 32, 65, 97, 324, 648, 972, 3239, 4859

97mg/ml for 5mg/plate

Vehicle / solvent:

- Solvent used: water
- Justification for choice of solvent: compatibility with the target cells

Test article was soluble in water at approximately 97 mg/ml, the maximum concentration tested.

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other:

Remarks:

All Salmonella Strains and WP2 uvrA (pKM101) with metabolic activation

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other:

Remarks:

WP2 (pKM101) without metabolic activation

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

TA98, TA1538 without metabolic activation

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

TA100, TA1535 without metabolic activation

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA1537 without metabolic activation

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

Both E.coli Strains without metabolic activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation
DURATION
- Preincubation period:
- Exposure duration:
- Expression time (cells in growth medium):
- Selection time (if incubation with a selection agent):
- Fixation time (start of exposure up to fixation or harvest of cells):

SELECTION AGENT (mutation assays):
SPINDLE INHIBITOR (cytogenetic assays):
STAIN (for cytogenetic assays):

NUMBER OF REPLICATIONS:

NUMBER OF CELLS EVALUATED:

DETERMINATION OF CYTOTOXICITY
- Method: other: dissecting microscope

OTHER EXAMINATIONS:
- Determination of polyploidy:
- Determination of endoreplication:
- Other:

OTHER:

Evaluation criteria:

For the test article to be evaluated positive, it must cause a dose-related increase in the mean revertants per plate of at least one tester strain with a minimum of two increasing concentrations of test article. Data sets for strains TA1535, TA1537 and TA1538 were judged positive if the increase in mean revertants at the peak of the dose response is equal to or greater than three times the mean vehicle control value. Data sets for strains TA98, TA100 and WP2 uvrA (pKM101) and WP2 (pKM101) were judged positive if the increase in mean revertants at the peak of the dose response is equal to or greater than two times the mean vehicle control value.

Key result

Species / strain:

E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Key result

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

True negative controls validity:

not examined

Positive controls validity:

valid

Key result

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

True negative controls validity:

not examined

Positive controls validity:

valid

Key result

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

True negative controls validity:

not examined

Positive controls validity:

valid

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 examined

True negative controls validity:

not examined

Positive controls validity:

valid

Key result

Species / strain:

E. coli WP2 uvr A pKM 101

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: none
- Effects of osmolality: none
- Evaporation from medium: no
- Water solubility: Yes at approximately 97 mg/ml
- Precipitation:
- Other confounding effects: not applicable

RANGE-FINDING/SCREENING STUDIES: Yes

COMPARISON WITH HISTORICAL CONTROL DATA: Yes

ADDITIONAL INFORMATION ON CYTOTOXICITY:

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Interpretation of results (migrated information):
negative

Under the conditions of this study, test article hydroxypropylated .beta.-cyclodextrin was concluded to be negative in the Salmonella/Mammalian Microsome (Ames Test) and Esclzericlzia coli WP2 Mutagenesis Assay.

Executive summary:

The test article, hydroxypropylated .beta.-cyclodextrin, was tested according to OECD Guideline 471 in the bacterial reverse mutation assay using S.typhimurium tester strains TA98, TA100, TA1535, TAl537 and TA1538 and E. coli tester strains WP2uvrA(pKM101) and WP2 (pKM101) in the presence and absence of Aroclor-induced rat liver S9. The assay was performed in two phases, using the plate incorporation and preincubation methods. The first phase, the dose range-finding study, was used to establish the dose range for the mutagenicity assay. The second phase, themutagenicity assay (initial and confirmatory assays), was used toevaluate the mutagenic potential of the test article. The preincubation method was used only for the confirmatory assay. The dosing solutions were adjusted to compensate for the purity of the test article.

Water was selected as the solvent of choice based on compatibility with the target cells. The test article was soluble in water at approximately 97 mg/ml, the maximum concentration tested.

In the preliminary toxicity assay, the maximum dose tested was 5 mg per plate; this dose was achieved using a concentration of 97 mg/ml and a 50 µl plating aliquot. Neither precipitate nor appreciable toxicity was observed. Based on the findings of the toxicity assay, the maximum dose plated in the mutagenicity assay was 5 mg per plate.

In the mutagenicity assay, no positive response was observed. Neither precipitate nor appreciable toxicity was observed.

Under the conditions of this study, test article hydroxypropylated .beta.-cyclodextrin was concluded to be negative in the Salmonella/Mammalian Microsome (Ames Test) and Esclzericlzia coli WP2 Mutagenesis Assay.

Reference 2

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

GLP compliance:

yes

Type of assay:

in vitro mammalian chromosome aberration test

Species / strain / cell type:

Chinese hamster Ovary (CHO)

Metabolic activation:

with and without

Metabolic activation system:

S-9 mix

Test concentrations with justification for top dose:

79, 157, 313, 625, 1250, 2500, and 5000 µg/mL in non-activated and activated

Vehicle / solvent:

- Solvents used: Ham's F12 medium supplemented with 10% fetal bovine serum, 2mM L-glutamine, 100 units penicillin and 100 µg streptomycin/ml
- Justification for choice of solvent: solibility

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

S9 activated

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: N-methyl-N'-nitro-N-nitrosoguanidine

Remarks:

S9 non-activated

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Preincubation period:
- Exposure duration:
- Expression time (cells in growth medium):
- Selection time (if incubation with a selection agent):
- Fixation time (start of exposure up to fixation or harvest of cells):

SELECTION AGENT (mutation assays):
SPINDLE INHIBITOR (cytogenetic assays):
STAIN (for cytogenetic assays):

NUMBER OF REPLICATIONS:

NUMBER OF CELLS EVALUATED: number and types of aberrations found are presented for each treatment flask

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other:

OTHER EXAMINATIONS:
- Determination of polyploidy:
- Determination of endoreplication:
- Other:

OTHER:

Evaluation criteria:

Prior to the analysis of the coded slides 50-100 cells from the solvent and positive control cultures were analyzed to determine that the percentage of aberrant cells in the solvent control \vas less than 6%, no cell had >2 aberrations and that the percentage of aberrant cells in the positive control was significantly increased relative to the solvent control.

Species / strain:

Chinese hamster Ovary (CHO)

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

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: none
- Effects of osmolality: none
- Evaporation from medium: no
- Water solubility: not critical
- Precipitation: no
- Other confounding effects: none found

RANGE-FINDING/SCREENING STUDIES: yes

COMPARISON WITH HISTORICAL CONTROL DATA: yes

ADDITIONAL INFORMATION ON CYTOTOXICITY: not available

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Interpretation of results (migrated information):
negative

Based on the findings of this study, hydroxypropylated .beta.-cyclodextrin was concluded to be negative for the induction of structural and numerical chromosome aberrations both in the presence and absence of S9 in Chinese hamster ovary (CHO) cells.

Executive summary:

The test article, hydroxypropylated .beta.-cyclodextrin , was tested according to OECD Guideline 473 in the chromosome aberration assay both in the absence and presence of metabolic activation (Aroclor-induced rat liver S9) using Chinese hamster ovary cells. The preliminary toxicity assay was conducted to establish the dose range for testing in the chromosome aberration assay. The chromosome aberration assay was conducted to evaluate the clastogenic potential of the test article. The test article dosing solutions were adjusted for the purity of the test article.

Culture medium (Ham's FI2 medium supplemented with 10% fetal bovine serum, 2mM L-glutamine, 100 units penicillin and 100 µg streptomycin/ml) was selected as the solvent of choice based on the solubility of the test article, information provided by the Sponsor and compatibility with the target cells. The test article was soluble in culture medium at approximately 500 mg/ml, the maximum concentration tested.

In the preliminary toxicity assay, the maximum dose level tested was 5000 µ/mL. The test article was soluble in treatment medium at all concentrations tested. No substantial toxicity, i.e., at least 50% cell growth inhibition, was observed at any of the doses tested regardless of exposure or harvest time. In the absence of substantial toxicity, the dose levels selected for testing in the chromosome aberration assay were 79, 157, 313, 625, 250, 2500, and 5000 µg/mL.

In the chromosome aberration assay, dose levels of 1250, 2500, and 5000 µg/mL were selected for microscopic analysis in both the non-activated and in the S9-activated test systems. Selection of doses for microscopic analysis was based on toxicity (the lowest dose with at least 50% reduction in cell growth and the next two lower doses) in all harvests.

Based on the findings of this study, hydroxypropylated .beta.-cyclodextrin was concluded to be negative for the induction of structural and numerical chromosome aberrations both in the presence and absence of S9 in Chinese hamster ovary (CHO) cells.

 

Reference 3

Endpoint:

in vitro gene mutation study in mammalian cells

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

Structural similarity

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

Chinese hamster Ovary (CHO)

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

Conclusions:

The substance is non mutagenic in mammalinan cells.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available