Cycloheptapentylose

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:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study report

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

see Principles

Principles of method if other than guideline:

The strains subcultured from storage dishes have been cultivated at 37°C in a nutritive medium (oxoid No 2 CM67). They have been collected in the morning from the shelf (approx. 2 x 10 x e9 bacteria/ml) and used the same day after approx. 16 hours of culture. The minimum medium used to evi-dence the mutants His+ is the V.B. medium.
The spreadings have been made with an overlayer containing a concentra-tion of 0.6 .% agar oxoid, 0.5 % sodium chloride, 0.05 mM biotine and L- Histidine, for a total volume of 2.5 ml. The amount of histidine has been cal-culated so as to allow the bacteria to divide 2 or 3 times on the plates since some mutagene agents operate only on growing cells.
A 500 mg/kg dose of Araclar 1254 dissolved in maize oil has been injected to 5prague-Oawley rats, through the peritoneum. On the 5th day after injec-tion, the rats were slaughtered. Livers were taken and homogenized at 4 •C in 0,15 M potassium chloride, and the mix was centrifuged 20 minutes at 9000 g. After centrifugation, the supernatant which constitutes the 59 frac-tion was kept in liquid nitrogen in 2 ml fractions.
The product Lab 870 has been solubilized in water for injection (Aguettant, batch 7428) at the concentration of 20 mg/ml. Successive dilutions have been carried out in the same solvent at following concentrations 10 - 5 - 1 - 0.1 mg/ml. The solutions and the pure product have been kept at 5°C for the trial duration.

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Target gene:

not applicable

Species / strain / cell type:

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

Additional strain / cell type characteristics:

not applicable

Species / strain / cell type:

S. typhimurium TA 1538

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

Pretest: 0.01 - 0.05 - 0.1 - 0.5 - 1 - 2 -4 mg /plate
Test: 0.1 - 0.5 - 1 - 2 - 4 mg/plate.

Vehicle / solvent:

water for injection

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: amino-2-anthracene (2-anthramine)

Remarks:

all strains with S9

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 S9

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA1537 without S9

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

TA100 without S9

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

ethylmethanesulphonate

Remarks:

TA1535 without S9

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation);

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3



DETERMINATION OF CYTOTOXICITY
- Method: mitotic index

Evaluation criteria:

None of the values obtained in presence of the product tested has been higher than, or equal to twice the value obtained in the presence of a soLvent with and without metabol ic activation on the 5 bacterial strains used.

Statistics:

not reported

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:

not applicable

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: not reported
- Effects of osmolality: not reported
- Evaporation from medium: not reported
- Water solubility: not reported
- Precipitation: not reported
- Other confounding effects: not reported


RANGE-FINDING/SCREENING STUDIES: yes


COMPARISON WITH HISTORICAL CONTROL DATA: not reported


ADDITIONAL INFORMATION ON CYTOTOXICITY: not reported

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Interpretation of results (migrated information):
negative

Substance is not mutagenic under the conditions used.

Executive summary:

The substance Beta-Cyclodextrin (LAB 870) has been tested on the 5 strains of Salmonella typhimurium (TA 98, TA 100, TA 1535, TA 1537, TA 1538) with and without metabolic activation (S9 -mix) according to the protocol of Ames and similar to OECD Guideline 471. A range of non-toxical concentrations had been determined in a preliminary experimentation on the strain TA 100 without metabolic activation. The 5 concentrations chosen (0.1, 0.5, 1, 2 and 4 mg/plate)have been tested in triplicate on the 5 strains quoted above, with and without metabolic activation. The results were confirmed in a second experiment separately from the first one.In each test a negative control (solvent) and a positive one (specific standard mutagene) had been included. The values obtained were consistent with the standards. Under these circumstances, the product did not show any mutagene power towards thestrains TA98, TA 100, TA 1535, TA 1537, TA .1538 with and without metabolic activation.

Reference 2

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study report, GLP compliant

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Deviations:

no

GLP compliance:

yes

Type of assay:

in vitro mammalian chromosome aberration test

Target gene:

not applicable

Species / strain / cell type:

lymphocytes: human

Details on mammalian cell type (if applicable):

- Type and identity of media:
- Properly maintained: yes

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

With metabolic activation 100 - 300 -1000 µg/ml
Without metabolic activation 100 - 300 - 1000 µg/ml

Vehicle / solvent:

RPMI 1640

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

cyclophosphamide

Remarks:

with metabolic activation

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

mitomycin C

Remarks:

without metabolic activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium;

DURATION

- Exposure duration: 1 hr (with metabolic activation), 24 hr (without metzabolic activation)
- Expression time (cells in growth medium): 22 hr
- Fixation time (start of exposure up to fixation or harvest of cells): 25 hr (with metabolic activation), 48 hr (without metzabolic activation)


SPINDLE INHIBITOR (cytogenetic assays): colcemide
STAIN (for cytogenetic assays): Giemsa


NUMBER OF REPLICATIONS: 2


NUMBER OF CELLS EVALUATED: 200


DETERMINATION OF CYTOTOXICITY
- Method: mitotic index


OTHER EXAMINATIONS:
- Determination of polyploidy: yes

Evaluation criteria:

A stUdy is accepted if :
- the mean number of cells carrying aberrations (excluding cells presenting gaps only) for the two subjects is less than 5 %·,
- the number of breaks per cell and number of cells with aberrations (excluding those with gaps only) is significantly increased.
- at least 3 of the cultures treated with the test compound present a mean mitotic index for the 2 subjects greater than about 50 % that found for the reference culture.
A compound is found to demonstrate clastogenicity against cultured human Iymphocytes if it results. ina statistically significant increase in the number of breaks per cell compared with the control or in the number of cells with aberrations (excluding gaps), if this increase amounts to at least a doubling of the control value and if the genotoxicity detected shows a.dose-effeet relationship.
A compound is found to have no clastogenic effect on cultured human Iymphocytes if it does not comply with any of the 3 criteria listed above.
If neither situation occurs, the results are discussed case by case and another independent study may be envisaged after modifying the dose range taking account of the findings obtained using other test systems.

Statistics:

Two types (gaps and breaks) are analyzed using the Student's t-test, because although the standard deviations found not to demonstrate that the normal distribution is normal it can be used because of the large number of cells (200 cells)
NUMERIC ABERRATIONS.: All numeric aberrations (aneuploidy and polyploidy) are analyzed statistically using the Chi2 test.
THE NUMBER OF DAMAGED CELLS WITH AND WITHOUT GAPS : The total number is analyzed statistically using the Chi2 test. From the comments already made regarding the significance of the presence of gaps, it results that greater weight is given to change in the number of cells presenting aberrations other than gaps in
assessing the genotoxicity·of the test compound.

Species / strain:

lymphocytes: human

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

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: not reported
- Effects of osmolality: not reported
- Evaporation from medium: not reported
- Water solubility: not reported
- Precipitation:not reported
- Other confounding effects: not reported


RANGE-FINDING/SCREENING STUDIES: yes


COMPARISON WITH HISTORICAL CONTROL DATA: Yes, compliant


ADDITIONAL INFORMATION ON CYTOTOXICITY:
With metabolic activation 10-30-100 - 300 -1000 µg/ml for 1 hr
Without metabolic activation 10-30-100 - 300 - 1000 µg/ml for 24 hr

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Interpretation of results (migrated information):
negative

Substance did not induce gaps or breaks exceeding the relevant values in this Chromosome aberration test.

Executive summary:

Beta-Cyclodextrin (purity not stated) was assessed in an in vitro Chromosome Aberration assay with human lymphocytes similar to the OECD guideline 473. The substance is soluble in aqueous medium, solutions were prepared in RPMI 1640 at the maximum concentration generally used at the laboratory, ie., 10 mg/ml. This solution was added at 10 % in culture medium. The highest final

concentration was 1000 µg/ml. A Preliminary test on cytotoxicity showed a low cytotoxicity of beta-cyclodextrin both with and without metabolic activation, at the highest studied dose 1000 µg/ml, demonstrated by a weak mitotic index decrease. The tests were performed with and without metabolic activation (S9 -mix) at concentrations of 100, 300 and 1000 µg/ml with 1 (with metabolic activation) and 24 hr (without metabolic activation) exposition time. Solvent, negative and positive controls worked as expected. Reference clastogens (mitomycin C without metabolic activation and cyclophosphamide with metabolic activation by the S9 mix) were tested in order to demonstrate the sensitivity of the cells and the effectiveness of the metabolic activation system. Both tests with and without metabolic showed no dose dependent positive results presents and thus the test substance did not show clastogenlc

activity in the in vitro human lymphocyte metaphase analysis test.

Reference 3

Endpoint:

in vitro gene mutation study in mammalian cells

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: Guideline study report, GLP compliant

Qualifier:

equivalent or similar to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Type of assay:

mammalian cell gene mutation assay

Target gene:

HPRT locus

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Details on mammalian cell type (if applicable):

- Type and identity of media:
- Properly maintained: yes

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

TOXICITY TEST
-Treatment time
With S9 mix: 3 hours
Without S9 mix: 3 hours
- Doses studied (µg/ml)
With S9 mix: 10 - 30 -100 - 300 - 1000
Without S9 mix: 10 - 30 -100 - 300 - 1000

MUTAGENICITY TEST
-Treatment time:
With S9 mix: 3 hours
Without S9 mix: 3 hours
- Doses studied (µg/ml)
With S9 mix: 30 -100 - 300 - 1000
Without S9 mix: 30 -100 - 300 - 1000

Vehicle / solvent:

H21 DMEM medium culture
Dulbecco H21 (Gibco) medium containing 7.5 % of foetal calf serum, glutamine (1 % of a 3 % solution) and antibiotics (Penicillin, Streptomycin,
Fungizone)

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

with S9

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

without S9

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium;

DURATION
- Exposure duration: 3 hr
-Expression time: T7
- Selection time (if incubation with a selection agent): 3 hr
- Fixation time (start of exposure up to fixation or harvest of cells):


SELECTION AGENT (mutation assays): 6-thioguanine (5 µg/Iml)
STAIN (for cytogenetic assays): Giemsa


NUMBER OF REPLICATIONS: 2


NUMBER OF CELLS EVALUATED:


DETERMINATION OF CYTOTOXICITY
- Method: mitotic index

Evaluation criteria:

A study is accepted if :
-the survival rate of the solvent control is 50 % higher than the number of the cells replated at TO and T7,
- the spontaneous mutation frequency of the negative control is tower than 2 x 10-5
- the mutation frequency of the positive control· is significantly increased compared with the solvent (higher 10 folds for EMS and higher 5 folds for B[a]P). The observed values must be close to those of historical positive controls.

Statistics:

not reported

Species / strain:

Chinese hamster lung fibroblasts (V79)

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

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: not reported
- Effects of osmolality: not reported
- Evaporation from medium: not reported
- Water solubility: not reported
- Precipitation: not reported
- Other confounding effects: not reported


RANGE-FINDING/SCREENING STUDIES: yes


COMPARISON WITH HISTORICAL CONTROL DATA: yes

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Interpretation of results (migrated information):
negative

Substance did not induce gene mutations in the HPRT-test

Executive summary:

A study equivalent to OECD 476 was performed to investigate the potential of beta-Cyclodextrin (100% pure) to induce gene mutations at the HPRT locus in V79 cells of the Chinese hamster. The assay was performed in two independent experiments, using two parallel cultures each. Both main experiments were performed with and without liver microsomal activation (S9) and a treatment period of 3 hours.The highest concentration of the test item was 1000 µg/mL. The tested concentrations were 30 -100 -300 and 1000µg/mL for both experiments with and without metabolic activation. No substantial and reproducible dose dependent increase of the mutation frequency was observed in both main experiments. Appropriate reference mutagens, used as positive controls, induced a distinct increase in mutant colonies and thus, showed the sensitivity of the test item and the activity of the metabolic activation system. In conclusion it can be stated that under the experimental conditions reported the test item did not induce gene mutations at the HPRT locus in V79 cells.Therefore, beta-Cyclodextrin is considered to be non-mutagenic in the HPRT assay.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Link to relevant study records

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study report, only one dose tested

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

yes

Remarks:

see principles

Principles of method if other than guideline:

one dose used (100 mg/kg)

GLP compliance:

yes

Type of assay:

micronucleus assay

Species:

mouse

Strain:

other: I.O.P.S., OFl (IFFA CREDO)

Sex:

male/female

Details on test animals or test system and environmental conditions:

The animals (6 - 8 weeks old mouse) were housed in groups of 2 (prelimi-nary study) or 5 (final study) in plastic cages (dimensions: 205 x 118 x 127 mm) on a dust free litter. The temperature was kept at 22 + 3 °C. The rela-tive air humidity was 30 - 70 %, with air changes of at least 8 per hour; arti-ficial lighting : 12 hours out of 24.
Commercial pelleted rat-mouse A04 CR diet - batch 70.630 (U.A.R. - Usine d'Alimentation Rationelle, Villemoisson sur Orge, France). Each manufac-turing batch was analysed to detect contaminants. Drinking water ad libitum.

Route of administration:

intraperitoneal

Vehicle:

carboxymethylcellulose hydrogel (1%)

Details on exposure:

one injection of 10 ml/kg

Duration of treatment / exposure:

24, 48, and 72 hours respectively

Frequency of treatment:

once

Post exposure period:

24, 48, and 72 hours respectively

Remarks:

Doses / Concentrations:
100 mg/kg bw
Basis:
nominal conc.

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Positive control(s):

cyclophosphamide
- Justification for choice of positive control(s):
- Route of administration:
- Doses / concentrations:

Tissues and cell types examined:

erythrocytes from the femur bone marrow

Details of tissue and slide preparation:

The animals were killed at the end of the treatment period by dislocation of the cervical vertebrae. The bone marrow removed from both femurs was suspended in foetal calf serum and centrifugated. For each animal, two smears were prepared on slides stained with May-Grunwald-Giemsa.
For each animal 1000 polychromatic erythrocytes were examined. A count of the number of polychromatophile erythrocytes bearing micronuclei was performed. At the same time, an observation and a count of the number of normochromatophile erythrocytes were carried out.
The results are expressed for each animal by the following ratio : number of normochromatophile erythrocytes for 400 polychromatophile erythrocytes. The average test and standard deviation were calculated for each group. The results are reported in the appendix.

Evaluation criteria:

not reported

Statistics:

The results are expressed as the number of cells bearing micronuclei for 1000 polychromatophile erythrocytes observed.
A statistical analysis wi th the aid of Student's test and the test of exact bilateral comparison were performed.

Sex:

male/female

Genotoxicity:

negative

Toxicity:

yes

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

RESULTS OF RANGE-FINDING STUDY
- Dose range: 100- 5000 mg/kg bw (intraperitoneal)
- Solubility: in vehicle soluble
- Clinical signs of toxicity in test animals: The results of the preliminary study show that the highest dose tolerated under the experimental conditions employed, averages 100 mg/kg. Therefore, the 100 mg/kg doses was chosen for the final trial.
- Evidence of cytotoxicity in tissue analyzed:
- Rationale for exposure: toxicity reasons, high mortality in higher concentrations
- Harvest times: 24, 48 and 72 h


RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): no
- Ratio of PCE/NCE (for Micronucleus assay): NCE/PCE 1.32 for 24 h harvest time, 0.82 (48 h) and 0.67 (72 h)
:

Conclusions:

Interpretation of results (migrated information): negative
Substance is not mutagenic in the in vivo micronucleus assay under the conditions used

Executive summary:

The mutagenic potential of the test article beta-Cyclodextrin (purity not specified) was tested in the I.O.P.S., OFl (IFFA CREDO) mouse using the micronucleus test, at a preliminarily defined dose of 100mg/kg according to a procedure similar to OECD 474.

The animals (5 males and 5 females per group) were administered the product by the intraperitoneal route (vehicle 1% carboxymethylcellulose hydrogel) and killed 24, 48 or 72 hours after administration. For each animal an examination of 1000 polychromatic erythrocytes obtained from the femoral bone marrow was performed. The statistical analysis of the results does not show any significant increase in the number of polychromatic erythrocytes bearing micronuclei in the animals treated with the test substance. The results obtained with the positive control (cyclophosphamide) are significantly positive. Concerning the ratio normochromatic/polychromatic erythrocytes which in the absence of toxical effect is close to 1, there was no significant increase in this value in the animals treated with the test article. A significant increase was observed in the animals treated with the positive control. To conclude, the test article did not induce any mutagenic effect in the mouse when administered at a dose of 100 mg/kg.

 

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

The substance Beta-Cyclodextrin has been tested on the 5 strains of Salmonella typhimurium (TA 98, TA 100, TA 1535, TA 1537, TA 1538) with and without metabolic activation (S9 -mix) according to the protocol of Ames and similar to OECD Guideline 471. A range of non-toxical concentrations had been determined in a preliminary experimentation on the strain TA 100 without metabolic activation.The 5 concentrations chosen (0.1, 0.5, 1, 2 and 4 mg/plate) have been tested in triplicate on the 5 strains quoted above, with and without metabolic activation. The results were confirmed in a second experiment separately from the first one.In each test a negative control (solvent) and a positive one (specific standard mutagene) had been included. The values obtained were consistent with the standards. Under these circumstances, the product did not show any mutagene power towards thestrains TA98, TA 100, TA 1535, TA 1537, TA .1538 with and withoutmetabolic activation.

A study equivalent to OECD 476 was performed to investigate the potential of beta-Cyclodextrin (100% pure) to induce gene mutations at the HPRT locus in V79 cells of the Chinese hamster. The assay was performed in two independent experiments, using two parallel cultures each. Both main experiments were performed with and without liver microsomal activation (S9) and a treatment period of 3 hours.The highest concentration of the test item was 1000 µg/mL. The tested concentrations were 30 -100 -300 and 1000µg/mL for both experiments with and without metabolic activation. No substantial and reproducible dose dependent increase of the mutation frequency was observed in both main experiments. Appropriate reference mutagens, used as positive controls, induced a distinct increase in mutant colonies and thus, showed the sensitivity of the test item and the activity of the metabolic activation system. In conclusion it can be stated that under the experimental conditions reported the test item did not induce gene mutations at the HPRT locus in V79 cells. Therefore, beta-Cyclodextrin is considered to be non-mutagenic in the HPRT assay.

Beta-Cyclodextrin was also assessed in an in vitro Chromosome Aberration assay with human lymphocytes similar to the OECD guideline 473. The substance is soluble in aqueous medium, solutions were prepared in RPMI 1640 at the maximum concentration generally used at the laboratory, ie., 10 mg/ml. This solution was added at 10 % in culture medium. The highest final

concentration was 1000 µg/ml. A Preliminary test on cytotoxicity showed a low cytotoxicity of beta-cyclodextrin both with and without metabolic activation, at the highest studied dose 1000 µg/ml, demonstrated by a weak mitotic index decrease. The tests were performed with and without metabolic activation (S9 -mix) at concentrations of 100, 300 and 1000 µg/ml with 1 (with metabolic activation) and 24 hr (without metabolic activation) exposition time. Solvent, negative and positive controls worked as expected. Reference clastogens (mitomycin C without metabolic activation and cyclophosphamide with metabolic activation) were tested in order to demonstrate the sensitivity of the cells and the effectiveness of the metabolic activation system. Both tests with and without metabolic showed no dose dependent positive results presents and thus the test substance did not show clastogenlc

activity in the in vitro human lymphocyte metaphase analysis test.

The recessive lethal mutation inducing effect of the compound beta-cyclodextrin was studied in Drosophila melanogaster by sex-linked recessive lethal test SLRL. Newly hatched Oregon-R males were treated with 1,6; 8.0 and 16 mM of beta-cyclodextrin for 24 hours. Each F1 female represents one paternal X-chromosome, treated in the male gametes. The vials to breed the F2 generation each corresponds to one treated male gamete and for that reason never should contain more than one F1 female. The vials of the F2 generation are then individually inspected for the presence of wild type /round red eyes/ males. Spontaneous (control) mutation frequency was determined with untreated animals. Suitability of test system was controlled by the positive effect of methyl methanesulfonate (MMS) a well-known mutagenic agent. It can be stated that the compound under study did not increase the frequency of the sex-linked recessive lethal mutation above the spontaneous level in the Drosophila melanogaster.

The mutagenic potential of the test article beta-Cyclodextrin (purity not specified) was tested in the I.O.P.S., OFl (IFFA CREDO) mouse using the micronucleus test, at a preliminarily defined dose of 100mg/kg according to a procedure similar to OECD 474.

The animals (5 males and 5 females per group) were administered the product by the intraperitoneal route (vehicle 1% carboxymethylcellulose hydrogel) and killed 24, 48 or 72 hours after administration. For each animal an examination of 1000 polychromatic erythrocytes obtained from the femoral bone marrow was performed.The statistical analysis of the results does not show any significant increase in the number of polychromatic erythrocytes bearing micronuclei in the animals treated with the test substance. The results obtained with the positive control (cyclophosphamide) are significantly positive.Concerning the ratio normochromatic/polychromatic erythrocytes which in the absence of toxical effect is close to 1, there was no significant increase in this value in the animals treated with the test article. A significant increase was observed in the animals treated with the positive control.To conclude, the test article did not induce any mutagenic effect in the mouse when administered at a dose of 100 mg/kg

 

 

Short description of key information:
All in vivo (micronucleus test) and in vitro (chromosome aberration, HPRT, Ames) Tests performed are negative.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

The substance did not show any mutagenic or clastogenic potential in any of the tests performed. Therefore, no classification according to CLP is required.