4-O-α-D-glucopyranosyl-D-glucitol

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

publication

Title:

The genotoxic and teratogenic effects of maltitol in rats

Author:

Semir Canimoglu and Eyyup Rencuzogullari

Year:

2012

Bibliographic source:

Toxicology and Industrial Health, 29 (10) 935-943

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 475 (Mammalian Bone Marrow Chromosome Aberration Test)

Version / remarks:

The assay design was based on OECD TG 475 version of 21 July 1997 with modifications.
Only 4 animals (2/sex) were used per dose (instead of 5 of one sex required) and higher doses than recomended were used.
Based on the 1997 version of OECD TG 475, the Intraperitoneal route was used for treatment and 100 metaphases/animal were evaluated in the current study.
However, since 1997, a new version of the OECD TG 475 is available (adopted in 29 July 2016). In the current guideline, intraperitoneal route is no more recommended and 200 metaphases should be evaluated instead of 100 analyzed in the 1997 version.

Deviations:

yes

Remarks:

See above in "Version/remarks" section

GLP compliance:

no

Type of assay:

mammalian bone marrow chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

4-O-α-D-glucopyranosyl-D-glucitol

EC Number:

209-567-0

EC Name:

4-O-α-D-glucopyranosyl-D-glucitol

Cas Number:

585-88-6

Molecular formula:

C12H24O11

IUPAC Name:

4-O-α-D-glucopyranosyl-D-glucitol

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: from Sigma, Batch No. M8892
- Expiration date of the lot/batch: not detailed
- Purity test date: no detailed

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: not specified
- Stability under test conditions: not specified
- Solubility and stability of the test substance in the solvent/vehicle: not specified
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: not applicable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
Maltitol was dossilved in distilled water, no more details.

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Medical Sciences, Experimental Research and Application Centre of Cukurova university
- Age at study initiation: 12-16 weeks old
- Weight at study initiation: 187.45±1.22g
- Assigned to test groups randomly: not detailed
- Fasting period before study: not detailed
- Housing: Plastic cages (32x46x10 cm)
- Diet (e.g. ad libitum): not detailed
- Water (e.g. ad libitum): not detailed
- Acclimation period: not detailed

ENVIRONMENTAL CONDITIONS
Not detailed

IN-LIFE DATES: Not detailed

Administration / exposure

Route of administration:

intraperitoneal

Vehicle:

- Vehicle(s)/solvent(s) used: Distilled water
- Justification for choice of solvent/vehicle: no justification provided
- Concentration of test material in vehicle: not specified
- Amount of vehicle (if gavage or dermal): not applicable
- Type and concentration of dispersant aid (if powder): not detailed
- Lot/batch no. (if required): not specified

Details on exposure:

The rats were intraperitoneally treated with 2.5, 5 and 10 g/kg body weight (bw) concentrations of maltitol.

Duration of treatment / exposure:

6, 12 and 24 h treatment periods.

Frequency of treatment:

Single administration

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 other: g/kg body weight

Doses / concentrations 2

Dose / conc.:

2.5 other: g/kg body weight

Doses / concentrations 3

Dose / conc.:

5 other: g/kg body weight

Doses / concentrations 4

Dose / conc.:

10 other: g/kg body weight

No. of animals per sex per dose:

Four rats were used per dose (2 males and 2 females)

Control animals:

yes

yes, concurrent vehicle

Positive control(s):

Ethyl-Carbamate Urethane
- Justification for choice of positive control(s): not specified
- Route of administration: intraperitoneal route
- Doses / concentrations: 0.4 g/kg bw

Examinations

Tissues and cell types examined:

Femur bone marrow was sampled for each animal and metaphases were isolated in order to be assessed.

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: no justification was provided

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields):
The rats were intraperitoneally treated with 2.5, 5 and 10 g/kg body weight (bw) concentrations of maltitol for 6, 12 and 24 h treatment periods.

DETAILS OF SLIDE PREPARATION:
In order to arrest mitosis, colchicine (1 mg/kg bw, Sigma C9754) was injected intraperitoneally 2 h before the animals were killed by cervical dislocation. The femurs were stripped proximally,
and the bone marrow was aspirated in 4 ml of 0.9%NaCl (37°C). The suspension was centrifuged for 10 min at 1200 r/min, and the obtained bone marrow pellet was resuspended in 0.4%KCl at 37°C for 5 min and then fixed in cold methanol glacial acetic acid (3:1) for 20 min at room temperature. The treatment with a fixative was repeated two times. Then, the cells were spread on glass slides and air-dried. The slides were stained with Giemsa (5% in Sorensen buffer)for 15 min.

METHOD OF ANALYSIS:
One hundred well-spread metaphases per animal (totally 400 metaphases per group)were examined at 1000Xmagnification for the occurrence of the structural and numerical CA. The mitotic index (MI) was also determined by scoring 3000 cells fromeach animal.

Statistics:

The t test was used for the statistical significance of the percentage of abnormal cells, the structural and total CA and MI. Dose–response relationships were
determined from the correlation and regression coefficients of the percentage of abnormal cells, structural and total Chromosomes Aberrations and Mitotic Index.

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Maltitol did not induce the percentage of abnormal cells and the structural CAs in all the concentrations and treatment periods. In addition, maltitol did not decrease the MI, so it had no cytotoxic effect in the bone marrow cells of rats.
There was no dose-dependent effect in all the tested parameters. Maltitol did not cause numerical chromosome abnormalities.

Any other information on results incl. tables

Table1.The percentage of abnormal cells,structural chromosome aberrations and the MI in rat bone marrow cells treated with maltitol.

 

Test substance	Periods (hours)	Concentrations (g/kgbw)	Abnormalities B'	Abnormalitie B''	Percent of abnormal cells± SE	Structural CA ± SE(%)	Mitosis index ± SE (%)
Control	–	–	1	4	1.00+0.40	1.25+0.47	3.66+0.13
Urethane	6	0.4	6	27	7.75+1.79	8.25+1.93	3.16+0.43
Maltitol	6	2.5	6	1	1.50+0.64	1.75+0.75	4.81+0.58
	6	5	2	5	1.25+0.47	1.75+0.75	4.37+0.53
	6	10	6	2	2.00+0.70	2.00+0.70	5.37+0.13
Urethane	12	0.4	14	15	6.75+1.43	7.25+1.54	4.65+0.52
Maltitol	12	2.5	8	4	2.75+0.85	3.00+0.81	4.29+0.69
	12	5	6	3	2.00+0.70	2.25+0.75	5.30+0.40
	12	10	1	4	1.00+0.40	1.25+0.62	5.39+0.48
Urethane	24	0.4	7	34	8.25+1.10	10.25+1.65	3.44+0.46
Maltitol	24	2.5	8	6	3.00+0.91	3.50+1.04	4.20+0.50
	24	5	5	1	1.50+0.28	1.50+0.28	4.79+0.51
	24	10	6	4	2.25+1.03	2.50+1.19	3.68+0.23
MI: mitotic index; CA: chromosome aberration; SE: standard error.
aStructuralchromosomal aberrations: B': chromatid type, B'': chromosome type abnormalities.

 

 

Applicant's summary and conclusion

Conclusions:

Under the experimental conditions of the study, Maltitol, when administered to rats by intraperitoneal route, did not induce chromosome aberrations or cytotoxic effect on bone marrow cells. hence, the maltitol was negative in a chromosome aberration test in vivo.

Executive summary:

In this non GLP compliant study, the genotoxic and cytotoxic effects of the maltitol were investigated in the bone marrow cells of rats using the chromosome aberration (CA) test, according to method similar to OECD 475 guideline.

To reveal the genotoxicity and cytotoxicity of maltitol, rats were intraperitoneally administered 2.5, 5 and 10 g/kg body weight (bw) concentrations of maltitol for 6, 12 and 24 h treatment period. Ethyl carbamate was used as positive control condition. Colchicine was used and administered to rats 2 hours before sacrifice in order to arrest mitosis. After treatment, femurs were sampled and bone marrow prepared in order to evaluate the chromosome aberrations and mitotic index.

Maltitol did not induce the percentage of abnormal cells and the structural CAs in all the concentrations and treatment periods. In addition, maltitol did not decrease the MI, so it had no cytotoxic effect in the bone marrow cells of rats. There was no dose-dependent effect in all the tested parameters. Maltitol did not cause numerical chromosome abnormalities.

Under the experimental conditions of the study, Maltitol, when administered to rats by intraperitoneal route, did not induce chromosome aberrations or cytotoxic effect on bone marrow cells. hence, the maltitol was negative in a chromosome aberration test in vivo.