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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

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Diss Factsheets

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

In vitro gene mutation study in bacteria:

Negative with and without metabolic activation in a study performed on the registered substance itself (Reliability 2 key study; non GLP compliant; similar to OECD 471 test guideline; Takizawa et al., 1984).

In vitro gene mutation study in mammalian cells:

Negative with and without metabolic activation based on read-across data on the analogue substance Syrups, corn, hydrogenated (Reliability 2 read-across key study, non-GLP compliant; similar to OECD guideline 476).

In vitro cytogenicity / chromosome aberration study in mammalian cells:

Key data on the registered substance itself: negative without metabolic activation in an in vitro chromosome aberration test and an in vitro Sister Chromatid Exchanges (SCE) test (Reliability 2 key studies, non-GLP compliant; similar to OECD test guidelines 473 and 479 with deviations respectively). Although the tests were performed without metabolic activation, the cytogenic potential of the substance is considered to be assessed as adequate reliable data from in vivo cytogenicity tests are available (see "Description of key information" in the "Genetic toxicity in vivo" section below).

Based on the available data and the analogy approach applied, Maltitol is not considered to be genotoxic in vitro.

Link to relevant study records

Referenceopen allclose all

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
Study period:
1982
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study without detailed documentation
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Deviations:
no
GLP compliance:
no
Type of assay:
mammalian cell gene mutation assay
Species / strain / cell type:
mouse lymphoma L5178Y cells
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Test concentrations with justification for top dose:
Toxicity test = 4, 12, 37, 110, 330, or 1000 µg/mL (± S9)
Mouse lymphoma (Test 1) = 61, 91, 135, 202, 301, 449, 670, or 1000 µg/mL (± S9)
Mouse lymphoma (Test 2) = 27, 41, 61, 91, 135, 202, 301, or 449 µg/mL (± S9)
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
- cell culture medium
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: - Ethylmethane sulfonate (- S9) and 3-methyl cholanthrene (+ S9)
Species / strain:
mouse lymphoma L5178Y cells
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
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

No dose-dependent increases in the mutation frequencies were noted in either mouse lymphoma test.

Conclusions:
No cytotoxicity and no increases in the mutant frequency were noted at any concentration either in the absence or presence of metabolic activation.
Executive summary:

According to the study authors, Lycasin, when tested at 8 concentrations ranging from 27 to 1000 µg/mL in the presence and absence of metabolic activation, was equivocal in the mouse lymphoma test. However, as no dose-dependent increases in the mutation frequencies were noted and the laboratory's criteria for determining a positive response were not met, it is thought that the result of the mouse lymphoma test is negative.

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2006
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Version / remarks:
The method was equivalent to current OECD TG 473. However, method description was not very detailed. No use of metabolic activation system (justified by the existance of reliable in vivo data). Only 24 hours and 48 hours treatment period was used, while short time exposure is also required by current guideline
Deviations:
yes
Remarks:
See "Version/remarks" section
GLP compliance:
no
Type of assay:
in vitro mammalian chromosome aberration test
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 specified
- Purity test date: not specified

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 specified

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: maltitol was diluted with sterile twice-distilled water
- Preliminary purification step (if any): not specified
- Final dilution of a dissolved solid, stock liquid or gel: not detailed
- Final preparation of a solid: not detailed

FORM AS APPLIED IN THE TEST (if different from that of starting material)
Diluted in sterile twice distilled water
Species / strain / cell type:
lymphocytes: human peripheral lymphocytes
Details on mammalian cell type (if applicable):
CELLS USED
- Source of cells: four healthy donors
- Suitability of cells: not detailed
- Cell cycle length, doubling time or proliferation index: not detailed
- Sex, age and number of blood donors if applicable: 21-23 years, 2 males and 2 females, nonsmokers
- Whether whole blood or separated lymphocytes were used if applicable: whole blood
- Number of passages if applicable: not applicable
- Methods for maintenance in cell culture if applicable: in chromosome medium supplemented with bromodeoxyuridine
- Modal number of chromosomes: 46
- Normal (negative control) cell cycle time: not detailed

MEDIA USED
- Type and identity of media including CO2 concentration if applicable: 2.5 mL chromosome medium supplemented with 10 µg/mL bromodeoxyuridine. The cultures were incubated at 37°C
- Properly maintained: not detailed
- Periodically checked for Mycoplasma contamination: not detailed
- Periodically checked for karyotype stability: not detailed
- Periodically 'cleansed' against high spontaneous background: not detailed
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
Colchicine
Metabolic activation:
without
Test concentrations with justification for top dose:
The cells were treated with maltitol at 1.25, 2.5, and 5.0 mg/mL concentrations
In this study, the concentration of maltitol used (5.0 mg/mL) is the maximum dose reported to be nontoxic by Madle et al. (1993). Maltitol was water soluble at 5.0 mg/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: sterile twice distilled water
- Justification for choice of solvent/vehicle: Maltitol was water soluble at 5.0 mg/mL
Untreated negative controls:
yes
Negative solvent / vehicle controls:
no
Remarks:
No solvent control was used because maltitol was diluted with sterile twice-distilled water
Positive controls:
yes
Positive control substance:
mitomycin C
Details on test system and experimental conditions:
METHOD OF APPLICATION: in suspension
- Cell density at seeding (if applicable): not detailed

DURATION
- Preincubation period: Cells were incubated at 37°C for 72 hours before treatment
- Exposure duration: 24 or 48 hours
- Expression time (cells in growth medium): No expression time
- Selection time (if incubation with a selection agent): not applicable
- Fixation time (start of exposure up to fixation or harvest of cells): 24 or 48 hours after treatment

SELECTION AGENT (mutation assays): not applicable

SPINDLE INHIBITOR (cytogenetic assays): Colchicine

STAIN (for cytogenetic assays): 5% Giemsa

NUMBER OF REPLICATIONS: One replicate per donor per condition (4 replicates per condition)

METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED: The cells were harvested by 0.4% KCl as a hypotonic solution and methanol:glacial acetic acid (3:1) as fixative. The staining of air-dried slides was performed follow- ing the standard methods using a 5% Giemsa stain

NUMBER OF CELLS EVALUATED: 400 cells per condition

NUMBER OF METAPHASE SPREADS ANALYSED PER DOSE (if in vitro cytogenicity study in mammalian cells): 100 wells spread first division metaphases per donor per condition (4 donors), 400 metaphases spreads analyzed per dose

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index, replication index
- Any supplementary information relevant to cytotoxicity: a total 400 cells (100 cells from each donor) were scored for replication index (RI). The mitotic index (MI) was also determined by scoring 3000 cells from each donor
Evaluation criteria:
The number of CA was obtained by calculating the percentage of metaphases for each concentration and treatment period that showed structural and/or numerical alterations. CA was classified according to the ISCN (according to Paz-y-Mino et al., 2002 from Mitelman, 1995).
Statistics:
The significance between the percentages of total CA were determined using the t-test. Dose-response relationships were determined from the correlation and regression coefficients for the percentage of total CA
Key result
Species / strain:
lymphocytes: human peripheral lymphocytes
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Maltitol induced structural CA (without statistical significance) while no change in numerical CA was observed. In addition, maltitol did not decrease the MI. The most common type of abnormality observed was of the chromatid type. No dose-relation effect and no statistical effect were observed.

Maltitol did not change the pH and osmolality the medium. The osmolality was measured as 312 mOsm/kg in control and 308 mOsm in the maximum dose of maltitol (5.0 mg/mL).

Table 1: Frequency of CA and MI in cultured human lymphocytes treated with maltitol.a

 

Test substance

Periods(h)

Concentration (mg/mL)

Chromatidtype

Chromosome type

Total CA±SE

MI ± SE

Control

 -

 -

7

3

2.50 ± 0.64

5.12±0.73

MMC

24

0.25mg/mL

74

40

28.50±5.72

3.61±0.89

Maltitol

24

1.25

11

9

5.00±1.22

4.97±1.29

Maltitol

24

2.50

23

1

6.00±0.70*

5.09±1.04

Maltitol

24

5.00

12

7

4.75±0.85

5.28±1.22

MMC

48

0.25mg/mL

476

381

214.25±32.74

2.05±0.53

Maltitol

48

1.25

16

4

5.00±1.22

6.01±0.62

Maltitol

48

2.50

18

5

5.75±1.22

5.46±0.23

Maltitol

48

5.00

15

1

4.00±1.68

6.48±0.74

aAtotal 400 cells were scored for CA and 12,000 cells were scored for MI.

 

*p < 0.05.

 

Conclusions:
Under the experimental conditions of the study, exposure of human peripheral lymphocytes to maltitol without metabolic activation, did not induce statistical difference or dose-relation effect for chromosome aberrations. Hence, Maltitol was negative in an in vitro chromosome aberration test without metabolic activation.
Executive summary:

The aim of this publication was to investigate the cytogenic potential of the maltitol when applied to human peripheral lymphocyte without metabolic activation and according to a method equivalent to current OECD TG 473 (with deviations).

Human peripheral lymphocytes were used and were obtained from whole blood (from four healthy donors). Cells were incubated for 72 hours before treatment. Maltitol were incubated with cells at 1.25, 2.5, and 5 mg/mL for 24 and 48 hours. Mytomicin C was used as positive control. No metabolic activation system was used. The cells were treated with colchicine and harvested to evaluated chromosome aberrations and mitotic index.

Under the experimental conditions of the study, exposure of human peripheral lymphocytes to maltitol without metabolic activation, did not induce statistical difference or dose-relation effect for chromosome aberrations. Hence, Maltitol was negative in an in vitro chromosome aberration test without metabolic activation.

Endpoint:
genetic toxicity in vitro, other
Remarks:
Sister Chromatid Exchange
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2006
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 479 (Genetic Toxicology: In Vitro Sister Chromatid Exchange Assay in Mammalian Cells)
Version / remarks:
Method was not very detailed. No short treatment performed. Only number of evaluated cells was given, no information on replicates performed was specified.
GLP compliance:
no
Type of assay:
sister chromatid exchange assay in mammalian cells
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 specified
- Purity test date: not specified

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 specified

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing:
maltitol was diluted with sterile twice-distilled water
- Preliminary purification step (if any):
not specified
- Final dilution of a dissolved solid, stock liquid or gel:
not detailed
- Final preparation of a solid: not detailed

FORM AS APPLIED IN THE TEST (if different from that of starting material)
Diluted in sterile twice distilled water
Species / strain / cell type:
lymphocytes: human peripheral lymphocytes
Details on mammalian cell type (if applicable):
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material:
From Sigma, batch no. M8892
- Expiration date of the lot/batch: not specified
- Purity test date: not specified

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 specified

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing:
maltitol was diluted with sterile twice-distilled water
- Preliminary purification step (if any):
not specified
- Final dilution of a dissolved solid, stock liquid or gel:
not detailed
- Final preparation of a solid: not detailed

FORM AS APPLIED IN THE TEST (if different from that of starting material)
Diluted in sterile twice distilled water
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
Colchicine
Metabolic activation:
without
Test concentrations with justification for top dose:
The cells were treated with maltitol at 1.25, 2.5, and 5.0 mg/mL concentrations.
In this study, the concentration of maltitol used (5.0 mg/mL) is the maximum dose reported to be nontoxic by Madle et al. (1993). Maltitol was water soluble at 5.0 mg/mL.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: sterile twice distilled water
- Justification for choice of solvent/vehicle: Maltitol was water soluble at 5.0 mg/mL
Untreated negative controls:
yes
Positive controls:
yes
Positive control substance:
mitomycin C
Details on test system and experimental conditions:
METHOD OF APPLICATION: in suspension
- Cell density at seeding (if applicable): not specified

DURATION
- Preincubation period: Cells were incubated at 37°C for 72 hours before treatment
- Exposure duration: 24 or 48 hours
- Expression time (cells in growth medium):No expression time
- Selection time (if incubation with a selection agent): not applicable
- Fixation time (start of exposure up to fixation or harvest of cells): 24 or 48 hours after treatment

SELECTION AGENT (mutation assays):
not applicable

SPINDLE INHIBITOR (cytogenetic assays): Colchicine

STAIN (for cytogenetic assays): modified fluorescence plus Giemsa

NUMBER OF REPLICATIONS: One replicate per donor per condition (4 replicates per condition)

METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED:
The cells were harvested by 0.4% KCl as a hypotonic solution and methanol:glacial acetic acid (3:1) as fixative. The staining of air-dried slides was performed following the standard methods using a modified fluorescence plus Giemsa method for SCE

NUMBER OF CELLS EVALUATED:
100 cells per condition

NUMBER OF METAPHASE SPREADS ANALYSED PER DOSE (if in vitro cytogenicity study in mammalian cells): In order to score SCE, 25 second- division metaphases were analyzed per sample.

DETERMINATION OF CYTOTOXICITY
- Method: replication index
- Any supplementary information relevant to cytotoxicity:
a total 400 cells (100 cells from each donor) were scored for replication index (RI).
Evaluation criteria:
The scoring of SCE was carried out according to the IPCS guidelines as described by Carrano and Natarajan (1988).
Statistics:
The significance between the mean SCE and RI (Replication Index) in treated cultures and their controls were determined using the t-test. Dose-response relationships were determined from the correlation and regression coefficients for the mean SCE and MI.
Key result
Species / strain:
lymphocytes: human peripheral lymphocytes
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Maltitol did not increase the SCE frequency at all concentrations and treatment times and it did not affect the RI.

Maltitol did not change the pH and osmolality the medium. The osmolality was measured as 312 mOsm/kg in control and 308 mOsm in the maximum dose of maltitol (5.0 mg/mL).

Table 1: Frequency of SCE and RI in cultured human lymphocytes treated with maltitol*

Test substance

Treatment periods (h)

Concentrations (mg/mL)

Min-maxSCE

SCE/Cell± SE

M1

M2

M2

RI ± SE

Control

 -

 -

1-16

6.82±0.47

19

95

292

2.69±0.06

MMC

24

0.25mg/mL

6–64

39.57±2.50

121

190

89

1.75±0.07

Maltitol

24

1.25

1–14

6.33±0.52

65

127

208

2.35±0.12

Maltitol

24

2.50

1–29

7.42±0.80

79

137

184

2.26±0.16

Maltitol

24

5.00

2–17

6.77±0.59

96

131

173

2.19±0.20

MMC

48

0.25mg/mL

36–141

74.02±3.62

207

172

21

1.53±0.11

Maltitol

48

1.25

1–14

6.23±0.37

42

112

246

2.51±0.09

Maltitol

48

2.50

1–16

6.69±0.60

33

206

161

2.32±0.15

Maltitol

48

5.00

1–21

6.34±0.84

21

219

160

2.34±0.17

*A total 100 cells were scored for the SCE assay; 400 cells were scored for RI.

 

Conclusions:
Under the experimental conditions of the study, Maltitol did not increase the SCE frequency at all concentrations and treatment times nor did maltitol affect the RI. Hence, Maltitol was not considered to be genotoxic in a in vitro Sister Chromatid Exchanges (SCE) test without metabolic activation.
Executive summary:

The aim of this publication was to investigate the cytogenic potential of maltitol when applied to human peripheral lymphocyte according to a method equivalent to current OECD TG 479 (with deviations).

Human peripheral lymphocytes were used and were obtained from whole blood (from four healthy donors). Cells were incubated for 72 hours before treatment. Maltitol were incubated with cells at 1.25, 2.5, and 5 mg/mL for 24 and 48 hours. Mytomicin C was used as positive control. No metabolic activation system was used. The cells were treated with colchicine and harvested to evaluated sister chromatid exchange and replication index.

Under the experimental conditions of the study, Maltitol did not increase the SCE frequency at all concentrations and treatment times nor did maltitol affect the RI. Hence, Maltitol was not considered to be genotoxic in a in vitro Sister Chromatid Exchanges (SCE) test without metabolic activation.

Endpoint:
in vitro gene mutation study in mammalian cells
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: read-across data with original study of reliability 2
Justification for type of information:
See attached document for read across analogy rationale and justification.
Reason / purpose for cross-reference:
read-across source
Key result
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
valid
Additional information on results:
No dose-dependent increases in the mutation frequencies were noted in either mouse lymphoma test.
Conclusions:
Under the experimental conditions of the study on the source substance, Syrups, hydrolyzed starch, hydrogenated did not induce mutagenicity in mammalian cells. Hence, based on the analogy approach, the target substance was not considered to be mutagenic for mammalian cells.
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1984
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
according to guideline
Guideline:
other: method of Nagao et al. which is essentially the same as the Ames Salmonella/mammalian microsome assay
Version / remarks:
Method not very detailed in the publication. Duplicates were used instead of triplicates as required by current standar methods. Escherichia coli Wp2/pKM101 was constructed by conjugation of TA98 and Wp2
Principles of method if other than guideline:
The same as the Ames Salmonella/mammalian microsome assay except for the preincubation for 20 min at 37 °C.
GLP compliance:
no
Type of assay:
bacterial reverse mutation assay
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Lot No. 212304
- Expiration date of the lot/batch: not specified
- Purity test date: not specified

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 specified

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Dissolved in distilled water
- Final dilution of a dissolved solid, stock liquid or gel: not specified
- Final preparation of a solid: dissolved in distilled water
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 mix obtained from Rat liver preparation was obtained from Aroclor 1254-treated male Sprague-Dawley rat
Test concentrations with justification for top dose:
0.5, 1.0, 2.0, 5.0, 10.0, 20.0 and 50.0 mg/plate
Justification for top dose not specified in the publication
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: distilled water
- Justification for choice of solvent/vehicle: not specified
Untreated negative controls:
yes
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
9-aminoacridine
2-nitrofluorene
N-ethyl-N-nitro-N-nitrosoguanidine
other: 2-aminoanthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 minutes
- Exposure duration: 48 hours
- Expression time (cells in growth medium): 48 hours
- Selection time (if incubation with a selection agent): not applicable
- Fixation time (start of exposure up to fixation or harvest of cells): 48 hours agter treatment

NUMBER OF REPLICATIONS: duplicates were ued per condition

METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED: not applicable

NUMBER OF CELLS EVALUATED: not applicable

DETERMINATION OF CYTOTOXICITY
- Method: not detailed
Evaluation criteria:
not detailed
Statistics:
not detailed
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
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:
valid
Positive controls validity:
valid
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
Positive controls validity:
valid
Additional information on results:
Neither maltitol (Maltitol crystal), nore Hydrogenated glucose syrups (Malti-Towa) induced an increase in the number of revertant colonies. The absence of the mutagenicity was shown not only in the standard Ames strains, Salmonella typhimurium TA1538, TA98, TA1537, TA1535, and TA100, but also in Escherichia coli WP2/pKM101 with or without S9 mix. No bactericidal effect was apparent in either test items, even at the maximal dose of 50 mg per plate.

Table 1 :results for Maltitol

Number of revertants per plate

 

Dose per plate (mg)

TA1538

 

TA98

 

TA1537

 

TA1535

 

TA100

 

WP2/pKM101

 

 

 - S9

 + S9

 - S9

 + S9

 - S9

 + S9

 - S9

 + S9

 - S9

 + S9

 - S9

 + S9

Control

0

16

35

21

37

6

12

28

9

155

171

150

253

Maltitol

0,5

11

33

22

37

6

14

24

10

168

166

150

246

 Maltitol

1

14

26

26

42

5

15

20

13

138

179

140

262

 Maltitol

2

18

34

17

37

6

11

22

11

156

154

152

271

Maltitol 

5

15

35

27

35

5

17

25

9

160

174

154

258

 Maltitol

10

14

25

23

39

5

11

30

9

165

180

152

267

 Maltitol

20

16

32

25

41

6

11

21

10

144

172

164

246

Maltitol 

50

16

26

23

41

5

17

18

11

152

172

148

264

Positive control

 

 

 

 

 

 

2-nitrofluorene

5 µg

2928

 

910

 

 

 

 

 

 

 

 

 

9-aminoacrimidine

10 µg

 

 

4428

 

 

 

 

4-nitroquinoline-1-oxide

0.5 µg

 

 

 

32

5939

1614

 

N-ethyl-N'-nitro-N-nitrosoguanodine

10 µg

 

 

 

323

3267

 

 

2-aminoanthracene

2 µg

 

1208

 

1239

 

61

 

501

 

2676

 

3074

 

Table 2 :results for Hydrogenated glucose syrups" 'Malti-Towa'

Number of revertants per plate

 

Dose per plate (mg)

TA1538

 

TA98

 

TA1537

 

TA1535

 

TA100

 

WP2/pKM101

 

 

 - S9

 + S9

 - S9

 + S9

 - S9

 + S9

 - S9

 + S9

 - S9

 + S9

 - S9

 + S9

Control

0

16

35

21

37

6

12

28

9

155

171

150

253

Malti-Towa

0,5

17

31

20

36

6

10

23

12

135

176

150

253

Malti-Towa 

1

17

35

24

35

7

16

21

10

155

167

144

259

Malti-Towa 

2

20

27

23

34

6

12

23

10

152

149

145

241

Malti-Towa 

5

16

36

26

37

5

11

24

11

150

161

143

252

Malti-Towa 

10

16

28

14

36

5

14

29

9

141

157

163

279

Malti-Towa 

20

20

39

19

34

5

15

24

12

146

159

127

262

 Malti-Towa

50

21

42

21

38

6

13

25

9

147

171

154

245

Positive control

 

 

 

 

 

 

2-nitrofluorene

5 µg

2928

 

910

 

 

 

 

 

 

 

 

 

9-aminoacrimidine

10 µg

 

 

4428

 

 

 

 

4-nitroquinoline-1-oxide

0.5 µg

 

 

 

32

5939

1614

 

N-ethyl-N'-nitro-N-nitrosoguanodine

10 µg

 

 

 

323

3267

 

 

2-aminoanthracene

2 µg

 

1208

 

1239

 

61

 

501

 

2676

 

3074

 

Conclusions:
Under the experimental conditions of the study, the test item Maltitol (Maltitol Crystal) did not induce mutagenicity in the bacterial strains tested both with and without metabolic activation. Hence, the test substance was not mutagenic in a reverse mutation bacterial assay.
Executive summary:

The aim of this non GLP is to evaluate the mutagenic potential of the test substance Maltitol Crystal (Maltitol, CAS 585 -88 -6) with a method based on the Ames method (Ames et al, 1975).

The test item Maltitol crystal was applied at concentrations from 0.5 mg to 50 mg/ plate to bacteria strain : TA98, TA100, TA1535, TA1537, TA1538 and E. Coli WP2/pKM101 under pre incubation method.

Bacteria were pre incubated for 20 minutes at 37°C and incubated in molten agar for 48 hours. Treatments were performed with and without metabolic activation system. Duplicates were used per condition.

Maltitol did not induce an increase in the number of revertant colonies. The absence of the mutagenicity was shown not only in the standard Ames strains, Salmonella typhimurium TA1538, TA98, TA1537, TA1535, and TA100, but also in Escherichia coli WP2/pKM101 with or without S9 mix. No bactericidal effect was apparent, even at the maximal dose of 50 mg per plate.

Under the experimental conditions of the study, the test item Maltitol (Maltitol Crystal) did not induce mutagenicity in the bacterial strains both with and without metabolic activation. Hence, the test substance was not mutagenic in a reverse mutation bacterial assay.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

- Maltitol was negative in a Bone Marrow Chromosomal Aberration Test in the rat via the intraperitoneal route (Reliability 2 key study; non-GLP compliant; similar to OECD 475; Canimoglu S and Rencuzogullari E. 2013).

- Maltitol was negative in an in vivo micronucleus test in the mouse via the oral route (Reliability 2 key study; non-GLP compliant; similar to OECD 474; Takizawa Y et al. 1984).

Link to relevant study records

Referenceopen allclose all

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
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
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.
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
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
Dose / conc.:
0 other: g/kg body weight
Dose / conc.:
2.5 other: g/kg body weight
Dose / conc.:
5 other: g/kg body weight
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
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.
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.

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.

 

 

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.

Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1984
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Version / remarks:
Study conducted one year after the adoption of the first version of OECD TG 474 (i.e. 1983). Methodology not very detailed in the publication.
Aqueous solution volume of 25mL/kg instead of the maximum of 20 ml/kg; No justification for the choice of doses specified.
1000 PCE/animal (as specified in 1983 version OECD TG 474) scored instead of 4000 as required by the latest version of OECD TG 474 (2016).
Deviations:
yes
Remarks:
see "Version / remarks"
GLP compliance:
no
Type of assay:
mammalian erythrocyte micronucleus test
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Lot No. 212304
- Expiration date of the lot/batch: not specified
- Purity test date: not specified

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 specified

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Dissolved in distilled water
- Final dilution of a dissolved solid, stock liquid or gel: not specified
- Final preparation of a solid: dissolved in distilled water
Species:
mouse
Strain:
other: ddY
Sex:
male
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Not detailed
- Age at study initiation: 6 weeks old
- Weight at study initiation: not specified
- Assigned to test groups randomly: not detailed
- Fasting period before study: not detailed
- Housing: not detailed
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: not detailed

ENVIRONMENTAL CONDITIONS
- Temperature (°C): standard condition, no more detail
- Humidity (%): standard condition, no more detail
- Air changes (per hr): standard condition, no more detail
- Photoperiod (hrs dark / hrs light): standard condition, no more detail

IN-LIFE DATES: not specified
Route of administration:
oral: gavage
Vehicle:
- Vehicle(s)/solvent(s) used: Distilled water
- Justification for choice of solvent/vehicle: no justification was provided
- Concentration of test material in vehicle: not detailed
- Amount of vehicle (if gavage or dermal): administration at 25mL/kg bw
- Type and concentration of dispersant aid (if powder): not detailed
- Lot/batch no. (if required): not detailed
- Purity:not applicable
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: Preparations were dissolved in distilled water

DIET PREPARATION
Not applicable
Duration of treatment / exposure:
The test material solution (25 ml/kg) was administered to 5 animals in each group by gastric intubation. Animals were killed following single or four repeated treatments, one dose per day for 4 days, within 24-30 h of the previous treatment.
Frequency of treatment:
Once a day, for one or four days
Post exposure period:
24-30 hours
Dose / conc.:
3 750 mg/kg bw/day (nominal)
Remarks:
One treatment
Dose / conc.:
7 500 mg/kg bw/day (nominal)
Remarks:
One treatment
Dose / conc.:
15 000 mg/kg bw/day (nominal)
Remarks:
One treatment
Dose / conc.:
7 500 mg/kg bw/day (nominal)
Remarks:
Four treatments, (equivalent to 30000 mg/kg bw)
No. of animals per sex per dose:
Male animals were used, at the number of 5 per experimental condition
Control animals:
yes
yes, concurrent vehicle
Positive control(s):
mitomycin C
- Justification for choice of positive control(s): no justification was provided
- Route of administration: intraperitoneal injection
- Doses / concentrations: 3mg/kg
Tissues and cell types examined:
Bonne marrow, erythrocytes
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION: Not detailed

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): Animals were sacrificed and sampled within 24-30 hours after the treatment. Femus were sampled and flushed.

DETAILS OF SLIDE PREPARATION: According to Yamato and Kikuchi method (1981) : One drop of new methylene blue staining solution consisting of 0.5 g of new methylene blue, 1.6 g of potassium oxalate, and 100 ml of distilled water, and the same amount of bone-marrow cell suspension were mixed and allowed to stand for 10 min at room temperature. Then the mixture was smeared, dried, fixed, and stained with Giemsa

METHOD OF ANALYSIS: The analysis of the slides was performed according to Schmid (1975); that is, the micronucleated erythrocytes were scored among 1000 polychromatic erythrocytes per mouse. Frequency of polychromatic erythrocytes was estimated in order to check whether the test compounds inhibit hemopoiesis.
Evaluation criteria:
Not detailed
Statistics:
The standard tables of Kastenbaum and Bowman (1970) were used fo the statistical analysis
Key result
Sex:
male
Genotoxicity:
negative
Toxicity:
no effects
Vehicle controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Both maltitol (Maltitol crystal) and Hydrogenated glucose syrups (Malti-Towa) revealed negative results concerning micronucleus induction. Neither an acute toxic effect on the animals nor hematopoiesis inhibition were observed in any of the experimental groups, with the exception of mitomycin C-treated mice (positive control group).

Table 1 :results for Maltitol

MicronucleatedPCE

 

Substance

Dose

Number of mice

Frequency of PCE

SD

Number of PCE observed

number

Frequency(%)

SD

Water

25 g/kg

5

57.9

6.6

5000

9

0.18

0.13

Maltitol

3750 mg/kgbw

5

56.8

3.1

5000

6

0.12

0.08

 

Maltitol

 

7500 mg/kgbw

5

51.1

3.7

5000

8

0.16

0.09

  

Maltitol

15000 mg/kgbw

5

56

5.3

5000

8

0.16

0.13

  

Maltitol

7500 mg/kg bw/day (4 days)

5

52

3.3

5000

11

0.22

0.04

MitomycinC

3 mg/kg

4

46.3

6

4000

177

4.43

0.55

 

Table 2 :results for Hydrogenated glucose syrups" 'Malti-Towa'

MicronucleatedPCE

 

Substance

Dose

Numberofmice

Frequencyof PCE

SD

Numberof PCEobserved

number

Frequency(%)

SD

Water

25 g/kg

5

57.9

6.6

5000

9

0.18

0.13

Malti-Towa

3750 mg/kgbw

5

64.1

6.4

5000

6

0.12

0.11

  

Malti-Towa

7500 mg/kgbw

5

54.1

8.4

5000

9

0.16

0.15

  

Malti-Towa

15000 mg/kgbw

5

61.2

7.6

5000

8

0.16

0.11

 

Malti-Towa

 

7500 mg/kg bw/day (4 days)

5

53.1

3.8

5000

6

0.12

0.13

MitomycinC

3 mg/kg

4

46.3

6

4000

177

4.43

0.55

 

Conclusions:
Under the experimental conditions of the study, Maltitol (Maltitol Crystal) did not induce an increase in the frequency of micronuclei when administered orally (gavage) to male mice. Hence, the test substance Maltitol was not considered as mutagenic in an in vivo micronucleus test.
Executive summary:

The aim of this non-GLP compliant in vivo study was to investigate the genotoxic potential of the Maltitol substance in mice, using a method equivalent to OECD TG 474.

The test material solution (25 ml/kg) was administered to 5 animals in each group by gastric intubation and animals were killed after single ( 0, 3750, 7500, 15000 mg/kg bw dose groups) or four repeated treatments (7500 mg/kg bw/day) of one dose per day for 4 days, within 24-30 h of the previous treatment.

Mitomycin C was used as positive control (administered intraperitoneally at 3mg/kg).

After sacrifice of animals, bone marrow was sampled for examination of micronucleuated polychromatic erythrocytes using stained slides. Micronucleated erythrocytes were scored among at least 1000 polychromatic erythrocytes per mouse. Frequency of polychromatic erythrocytes was estimated in order to check whether the test compound inhibit hemopoiesis.

Maltitol revealed negative results concerning micronucleus induction. Neither an acute toxic effect on the animals nor hematopoiesis inhibition were observed in any of the experimental groups, with the exception of mitomycin C-treated mice (positive control group).

Under the experimental conditions of the study, Maltitol (Maltitol Crystal) did not induce an increase in the frequency of micronuclei when administered orally (gavage) to male mice. Hence, the test substance Maltitol was not considered as mutagenic in an in vivo micronucleus test.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Genetic toxicity in vitro

A key non-GLP bacterial reversion mutation assay was performed with the registered substance Maltitol (Maltitol Crystal as reported in the study) (Reliability 2; Similar to Ames test; Takizawa Y et al, 1984). Salmonella typhimurium strains TA98, 100, 1535, 1537 and E. Coli WP2/pKM101 were exposed to concentrations up to 50 mg/plate, with and without metabolic activation. Maltitol induced no detectable revertants in any of the tester strains at doses of 0.5-50 mg per plate both with and without metabolic activation. Thus, under the experimental conditions of this test, Maltitol was not mutagenic in a bacterial reverse mutation test. In this same study, Hydrogenated glucose syrups (Malti-Towa as reported in the study) was also tested for bacterial mutation potential in the same conditions as Maltitol. The results showed that Malti-Towa was also not mutagenic in a bacterial reverse mutation test. A reliable non-GLP compliant, bacterial reverse mutation assay (Reliability 2; similar to OECD guideline 471), is also available on the analogue substance Syrups, corn, hydrogenated. The analogue was tested at doses of 0, 0.01, 0.05, 0.1, 0.5, and 1 mL/plate with the Salmonella typhimurium strains TA 98, TA 100, TA 1535, and TA 1537 both with and without metabolic activation. The substance was negative with and without metabolic activation, under the experimental conditions of this study.

The results of the in vitro gene mutation studies in bacteria on both the target and the source substance are indicative of a similar behavior regarding mutagenicity in bacterial systems.

Non-GLP cytogenicity tests were performed to assess the cytogenic potential of Maltitol in human peripheral lymphocytes (published data). Cells were exposed to the concentrations of 1.25, 2.5, and 5.0 mg/mL for 24h and 48h treatment times only without metabolic activation. The results of the Sister Chromatid Exchanges (SCE) test (Reliability 2; non GLP compliant; Similar to OECD 479 with deviations; Canimoglu S and Rencuzogullari E, 2006), showed that Maltitol did not induce sister chromatid exchanges at any of the tested doses or treatment periods (24 and 48h). In the chromosome aberration test (Reliability 2; non GLP compliant; Similar to OECD 473; Canimoglu S and Rencuzogullari E, 2006), Maltitol induced no change in numerical chromosome aberrations and induced structural chromosomal aberrations but without statistical significance. In addition, Maltitol did not decrease the replication index or the mitotic index at all doses and treatment periods, nor did it alter the pH or osmolality of the medium. The authors concluded that Maltitol has a weak genotoxic potential and appears to be non-cytotoxic to human peripheral lymphocytes in vitro. In this same published data (i.e. Canimoglu S and Rencuzogullari E, 2006), Maltitol was also tested in a micronucleus test without metabolic activation in human peripheral lymphocytes, in which maltitol induced micronucleus frequency but no dose-relation effect was observed.

However, due to a lack in the methodology description (in particular the handling and treatment of cells), the validity of this third test could not be assessed and this test was only reported as supporting with a reliability of 4. Although the cytogenicity tests on Maltitol were performed without metabolic activation, the cytogenic potential of the substance is considered to be assessed as adequate reliable data from in vivo cytogenicity tests are available (see the “In vivo Genotoxicity” section below).

A reliable, non-GLP mammalian chromosome aberration test (Reliability 2, similar to OECD guideline 473) is also available on the analogue substance Syrups, corn, hydrogenated. Chinese Hamster Ovary cells (CHO) were exposed to the substance at dose levels of 0 (control), 49, 164, 490, 1640, or 4900 μg/mL, both with and without metabolic activation. Syrups, corn, hydrogenated did not induce cytotoxicity or chromosome damage at any concentration. Hence, both the target substance and the analogue substance share a common weak cytogenic potential in mammalian cell systems.

No mammalian gene mutation data is available on Maltitol. A key, non-GLP mammalian gene mutation assay (Reliability 2, similar to OECD guideline 476) was conducted with the read-across substance Syrups, corn, hydrogenated. When the L5178Y cells were exposed to the substance at levels of 0 (control), 27, 41, 61, 91, 135, 202, 301,449, 670, or 1000 μg/mL, both with and without metabolic activation, no cytotoxicity and no increases in the mutant frequency were noted at any concentration either in the absence or presence of metabolic activation. Hence, under the experimental conditions of this study, Syrups, corn, hydrogenated was not mutagenic in a mammalian gene mutation test.

Genetic toxicity in vivo

Two in vivo key mutagenicity studies are available on the target substance Maltitol. A non-GLP reliable Bone Marrow Chromosomal Aberration Test (Reliability 2, similar to OECD 475 guideline with deviations; Canimoglu S and Rencuzogullari E. 2013) was performed. The genotoxic and cytotoxic effects of Maltitol were investigated in the bone marrow cells of rats. Rats were intraperitoneally administered 2.5, 5 and 10 g/kg body weight concentrations of Maltitol for 6, 12 and 24h. The results of this study showed that Maltitol did not induce chromosome aberrations and did not decrease the mitotic index in bone marrow cells of rats at all the tested doses and treatment periods.

The second key in vivo cytogenicity study is a non GLP in vivo micronucleus test (Reliability 2, similar to OECD 474 with deviations; Takizawa Y et al. 1984). Groups of male mice were orally administered Maltitol at the doses of 3.75, 7.5, 15 and 30 g/kg bw by gavage. Animals were killed following single or four repeated treatments, one dose per day for 4 days, within 24-30h of the previous treatment. A positive control group was included: 4 mice were intraperitoneally injected with 3 mg/kg bw of Mitomycin C. Bone marrow-smeared slides stained with Giemsa solution were prepared, analysed and the micronucleated erythrocytes were scored among 1000 polychromatic erythrocytes per mouse. The frequency of polychromatic erythrocytes was estimated in order to check whether the test compounds inhibit hemopoiesis. The results of this test showed that Maltitol induced no significant increase in the frequency of micronucleated erythrocytes in the bone marrow of mice at any of the tested doses. Based on both key studies, Maltitol is not considered to be mutagenic in vivo.

A reliable, non-GLP in vivo micronucleus test (Reliability 2, similar to OECD guideline 474) is also available on the read-across substance Syrups, corn, hydrogenated. Male mice were administered Syrups, corn, hydrogenated at doses of 0 (control), 5, or 25 mL/kg bw once/day for 2 days via the oral (gavage) route. Following sacrifice of the animals 6h after the final dose administration and subsequent examination of the bone marrow, no evidence of mutagenic potential or bone marrow cell toxicity at any concentration was noted. The only death reported was in one male of the high-dose group. Based on the results of this study, Syrups, corn, hydrogenated was not mutagenic in vivo. Thus, the target substance and the analogue substance share a common behavior regarding the in vivo genotoxicity endpoint.

The overall results of the in vitro and in vivo genotoxicity studies available on the source and the target substances are indicative of a similar behavior regarding the genetic toxicity endpoint. Hence, based on the data available on both the target and the read-across substance, the registered substance Maltitol is not considered to be genotoxic. This very similar behavior regarding the genotoxic potential further reinforces the read-across justification.

Justification for classification or non-classification

The available in vitro and in vivo genotoxicity data on the target substance Maltitol as well as on its analogue Syrups, corn, hydrogenated showed negative results. Based on the above, the substance is not considered to meet the criteria for classification according to Regulation (EC) No 1272/2008.