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Diss Factsheets

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

No genotoxicity study is available on the registered substance, however, the three required in vitro studies are available on an analogue substance (THFMA):
- negative results are obtained in the Ames test with S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 (met. act.: with and without) (similar to OECD TG 471; GLP);
- negative results are obtained in the Mammalian cell gene mutation assay with V79 cells (HPRT) (met. act.: with and without) (OECD TG 476; GLP);
- negative results are obtained in vitro mammalian cell micronucleus assay with cultured peripheral human lymphocytes (met. act.: with and without) (OECD TG 487; GLP).

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2013-09-04 - 2013-10-25
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
other: EU method B.49 (In vitro Mammalian Cell Micronucleus Test), dated July 06, 2012.
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 487 (In vitro Mammalian Cell Micronucleus Test)
Version / remarks:
adopted July 22, 2010
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
in vitro mammalian chromosome aberration test
Target gene:
not applicable
Species / strain / cell type:
lymphocytes: human, healthy donors not receiving medication
Details on mammalian cell type (if applicable):
- Type and identity of media: Dulbecco's Modified Eagles Medium/Ham's F12 (DMEM/F12, mixture 1:1) already supplemented with 200 mM GlutaMAX, supplemented with penicillin/streptomycin (100 U/mL/100 Lg/mL), the mitogen PHA (3 Lg/mL), 10 % FBS (fetal bovine serum), 10 mM HEPES and the anticoagulant heparin (125 U.S.P.-U/mL)
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: not relevant
- Periodically checked for karyotype stability: not relevant
- Periodically "cleansed" against high spontaneous background: not relevant
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
Experiment I (4 h exposure, 16 h recovery, fixation time 40 h, without S9): 11.0, 19.3, 33.8, 59.2, 103.6, 181.3, 317.2, 555.1, 971.4, 1700.0 µg/mL
Experiment I (4 h exposure, 16 h recovery, fixation time 40 h, with S9): 11.0, 19.3, 33.8, 59.2, 103.6, 181.3, 317.2, 555.1, 971.4, 1700.0 µg/mL

Experiment II (20 h exposure, fixation time 40 h, without S9): 11.0, 19.3, 33.8, 59.2, 103.6, 181.3, 317.2, 555.1, 971.4, 1700.0 µg/mL
Experiment II (4 h exposure, 16 h recovery, fixation time 40 h, with S9): 103.6, 181.3, 317.2, 555.1, 971.4, 1700.0 µg/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO, final concentration in medium 0.5%
- Justification for choice of solvent/vehicle: The solvent was chosen due to its solubility properties and its relative non-toxicity to the cell cultures.
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
mitomycin C
other: (without metabolic activation, pulse treatment)
Positive controls:
yes
Positive control substance:
other: Demecolcin (without metabolic activation, continuous treatment)
Positive controls:
yes
Positive control substance:
cyclophosphamide
other: (with metabolic activation)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium

DURATION
- Exposure duration:
Experiment I:
4 hours pulse treatment (with and without metabolic activation), expression phase 16 hours, cytokinesis block 20 hours.
Experiment II:
4 hours pulse treatment (with metabolic activation), expression phase 16 hours, cytokinesis block 20 hours
20 hours continuous treatment (without metabolic activation), cytokinesis block 20 hours
- Expression time (cells in growth medium): 20 h (+ cytochalasin B)
- Fixation time (start of exposure up to fixation or harvest of cells): 40

A series of in-house non-GLP validation experiments was performed to get distinct responses of statistical significance when using the specified positive controls. To achieve such response the test design, specifically for the treatment, the recovery phase and harvest time, was slightly modified comparing the current proposal given in the OECD Guideline 487.

SPINDLE INHIBITOR (cytogenetic assays): 4 µg/mL cytochalasin B
STAIN (for cytogenetic assays): Giemsa

NUMBER OF REPLICATIONS: 2 per experiment
NUMBER OF CELLS EVALUATED: 1000 binucleate cells per culture were scored for cytogenetic damage

DETERMINATION OF CYTOTOXICITY
- Method: CBPI (Cytokinesis-block proliferation index) was determined in approximately 500 cells per culture and cytotoxicity is expressed as % cytostasis
Evaluation criteria:
The micronucleus assay is considered acceptable if it meets the following criteria:
- The number of micronuclei found in the negative and solvent controls falls within the range of the laboratory historical control data
- The positive control substances should produce significant increases in the number of cells with micronuclei.

A test item can be classified as non-mutagenic if:
- the number of micronucleated cells in all evaluated dose groups is in the range of the laboratory historical control data and/or
-no statistically significant or concentration-related increase in the number of micronucleated cells is observed.
A test item can be classified as mutagenic if:
- the number of micronucleated cells is not in the range of the historical laboratory control data and
- either a concentration-related increase of micronucleated cells in three test groups or a statistically significant increase of the number of micronucleated cells is observed.
Statistics:
Chi square test (alpha < 0.05)
Species / strain:
lymphocytes: human
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: Exp. I: no cytotoxicity with and without S9; Exp II: cytotoxic without S9, no cytotoxicity with S9
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: 7.6 / 7.8 for solvent control, 7.7 / 7.8 for test item in experiment I / II -> no relevant influence
- Effects of osmolality: 376 / 385 for solvent control, 375 / 381 for test item in experiment I / II -> no relevant influence
- Precipitation: No precipitation of the test item in the culture medium was observed.

COMPARISON WITH HISTORICAL CONTROL DATA: all results were within the range of the laboratory historical control data

ADDITIONAL INFORMATION ON CYTOTOXICITY:
In Experiment I in the absence and presence of S9 mix and in Experiment II in the presence of S9 mix, no cytotoxicity was observed up to the highest applied concentration. In Experiment II in the absence of S9 mix the highest applied concentration showing clear cytotoxic effects was not evaluable.

 

 

Proliferation index CBPI, mean of 2 cultures

Cytostasis [%]

Micronucleated cells [%]

Experiment I; Exposure period 4 hrs without S9 mix

Solvent control, 0.5% DMSO

2.01

---

0.85

Positive control, 2.0 µg/mL MMC

1.70

30.8

15.50

Test item 555.1 µg/mL

1.95

5.4

0.60

Test item 971.4 µg/mL

1.77

24.0

0.80

Test item 1700.0 µg/mL

1.89

11.8

0.75

Experiment I; Exposure period 4 hrs with S9 mix

Solvent control, 0.5% DMSO

1.86

---

0.95

Positive control, 17.5 µg/mL CPA

1.87

n.c.

3.10

Test item 555.1 µg/mL

1.91

n.c.

0.35

Test item 971.4 µg/mL

1.92

n.c.

0.50

Test item 1700.0 µg/mL

1.83

3.5

0.45

Experiment II; Exposure period 20 hrs without S9 mix

Solvent control, 0.5% DMSO

1.82

---

0.50

Positive control, 125 ng/mLDemecolcin

1.47

43.2

5.55

Test item 317.2 µg/mL

1.88

n.c.

0.25

Test item 555.1 µg/mL

1.75

8.6

0.40

Test item 971.4 µg/mL

1.71

14.0

0.40

Experiment II; Exposure period 4 hrs with S9 mix

Solvent control, 0.5% DMSO

1.92

---

0.55

Positive control, 15 µg/mL CPA

1.70

23.7

3.10

Test item 555.1 µg/mL

1.94

n.c.

0.65

Test item 971.4 µg/mL

1.95

n.c.

0.70

Test item 1700.0 µg/mL

1.96

n.c.

0.60

 

n.c.= Not calculated as the CBPI is equal or higher than the solvent control value

 

 

Conclusions:
In this test under the experimental conditions reported, the test item Tetrahydrofurfuryl methacrylate did not induce micronuclei in human lymphocytes in vitro when tested up to cytotoxic or the highest evaluable concentration.
Executive summary:

In a mammalian cell micronucleus assay according to OECD guideline 478 (adopted July 22, 2010) and EU method B.49, dated July 06, 2012., primary human lymphocyte cultures were exposed toTetrahydrofurfuryl methacrylate(99.0%) in DMSO with and without metabolic activation (S9 mix).

The following concentrations were evaluated (calculations were not adjusted to purity):

 

Experiment I:

4 h exposure, 16 h recovery, fixation time 40 h, without S9: 555.1, 971.4, 1700.0 µg/mL

4 h exposure, 16 h recovery, fixation time 40 h, with S9: 555.1, 971.4, 1700.0 µg/mL

Experiment II:

20 h exposure, fixation time 40 h, without S9: 317.2, 555.1, 971.4 µg/mL

4 h exposure, 16 h recovery, fixation time 40 h, with S9: 555.1, 971.4, 1700.0 µg/mL

 

The test item was tested up to cytotoxic or the guideline limit concentration of 10 mM (corresponding to 1700.0 µg/mL). No cytotoxicity was observed in Experiment I with and without metabolic activation as well as in Experiment II with metabolic activation. In Experiment II in the absence of S9 mix the highest applied concentration showing clear cytotoxic effects was not evaluable.

In both independent experiments, neither a statistically significant nor a biologically relevant increase in the number of micronucleated cells was observed after treatment with the test item. 

Positive controls induced the appropriate response. 

There was no evidence of micronucleated cells induced over background. Therefore,Tetrahydrofurfuryl methacrylate is considered to be non-clastogenic in this in vitro micronucleus test, when tested up to cytotoxic or the guideline limit concentration.

NOTE: Any of data in this dataset are disseminated by the European Union on a right-to-know basis and this is not a publication in the same sense as a book or an article in a journal. The right of ownership in any part of this information is reserved by the data owner(s). The use of this information for any other, e.g. commercial purpose is strictly reserved to the data owners and those persons or legal entities having paid the respective access fee for the intended purpose.

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2006-12-19 to 2007-01-25
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay
Target gene:
his (Salmonella strains), trp (E. coli strain)
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
Experiment 1: 0, 4.88, 19.5, 78.1, 313, 1250, 5000 µg/plate
Experiment 2: 0, 156, 313, 625, 1250, 2500, 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
other: 2-Aminoanthracene
Remarks:
with metabolic activation
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
other: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide
Remarks:
without metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 2

DETERMINATION OF CYTOTOXICITY
- Method: reduced bacterial growth
Evaluation criteria:
Although not given in the study report, according to OECD guideline 471 a test item is considered mutagenic if:
- a clear and dose-related increase in revertant number occurs
- a biologically relevant positive response for at least one dose group occurs: in TA 100 and E. coli uvrA number of revertants at least twice as high as in solvent control; TA 98, TA 1535, TA 1537 at least three times higher number of revertants as in solvent control
Statistics:
no
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
valid
Additional information on results:
ADDITIONAL INFORMATION ON CYTOTOXICITY:
cytotoxic effects were observed in strains TA98, TA100, TA1535 and TA1537 at 5000 µg/plate without metabolic activation and in strains TA100 and TA1535 at 5000 µg/plate with metabolic activation

 

 

S9 mix

test item concentration [µg/plate]

mean number revertants/plate

TA100

TA1535

E. coli WP2uvrA-

TA98

TA1537

without

0

117

12

28

10

5

4.88

92

8

23

17

9

19.5

99

10

23

13

7

78.1

107

11

26

15

7

313

91

12

23

16

4

1250

109

10

20

17

5

5000

80*

8*

28

11*

5*

positive control

722

404

257

414

138

with

0

112

9

32

26

10

4.88

105

12

33

23

15

19.5

92

7

33

29

12

78.1

111

9

30

26

7

313

110

10

38

30

8

1250

124

7

34

24

10

5000

87*

11*

30

22

9

positive control

1341

256

1017

421

124

 

S9 mix

test item concentration [µg/plate]

mean number revertants/plate

TA100

TA1535

E. coli WP2uvrA-

TA98

TA1537

without

0

106

13

16

14

7

156

113

14

24

12

4

313

108

11

24

18

5

625

88

15

21

18

10

1250

110

13

20

12

7

2500

114

20

18

16

7

5000

65*

4*

18

10*

9*

positive control

679

454

188

520

195

with

0

112

10

26

26

12

156

109

5

28

21

11

313

99

8

28

29

10

625

104

7

30

26

11

1250

112

11

36

22

10

2500

110

12

33

21

12

5000

99*

10*

37

25

12

positive control

1311

276

981

426

132

 

* cytotoxic, reduced bacterial growth

Conclusions:
THFMA was not mutagenic in the bacterial reverse gene mutation assay when tested up to cytotoxic or limit concentrations (5000 µg/plate).
Executive summary:

In a reverse gene mutation assay in bacteria similar to OECD guideline 471, S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 strains were exposed to THFMA in DMSO at concentrations of 0, 4.88, 19.5, 78.1, 313, 1250, 5000 µg/plate in the first experiment and 0, 156, 313, 625, 1250, 2500, 5000 µg/plate in the second experiment in the presence and absence of mammalian metabolic activation (S9 mix).  THFMA was tested up to limit concentrations (5000 µg/plate).

Cytotoxic effects (reduced bacterial growth) were observed in strains TA98, TA100, TA1535 and TA1537 at 5000 µg/plate without metabolic activation and in strains TA100 and TA1535 at 5000 µg/plate with metabolic activation.

There was no evidence of induced mutant colonies over background. 

THFMA was not mutagenic in this bacterial reverse gene mutation assay when tested up to cytotoxic or limit concentrations (5000 µg/plate).

NOTE: Any of data in this dataset are disseminated by the European Union on a right-to-know basis and this is not a publication in the same sense as a book or an article in a journal. The right of ownership in any part of this information is reserved by the data owner(s). The use of this information for any other, e.g. commercial purpose is strictly reserved to the data owners and those persons or legal entities having paid the respective access fee for the intended purpose.

Endpoint:
in vitro gene mutation study in mammalian cells
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2013-09-10 - 2013-10-14
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
Version / remarks:
dated May 30, 2008
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Version / remarks:
adopted July 21, 1997
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
mammalian cell gene mutation assay
Target gene:
HPRT
Species / strain / cell type:
Chinese hamster lung fibroblasts (V79)
Details on mammalian cell type (if applicable):
- Type and identity of media: MEM (minimal essential medium) containing Hank’s salts supplemented with 10% foetal bovine serum (FBS), neomycin (5 µg/mL) and amphotericin B (1%)
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: yes
- Periodically "cleansed" against high spontaneous background: yes
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
Experiment I (4 h exposure, with and without S9): 53.8, 107.5, 215.0, 430.0, 860.0, 1720.0 µg/mL

Experiment II (24 h exposure, without S9): 107.5, 215.0, 430.0, 860.0, 1290.0, 1720.0 µg/mL
Experiment II (4 h exposure, with S9): 107.5, 215.0, 430.0, 860.0, 1290.0, 1720.0 µg/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO, final concentration in medium 0.5%
- Justification for choice of solvent/vehicle: The solvent was chosen due to its solubility properties and its relative non-toxicity to the cell cultures.
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
ethylmethanesulphonate
other: (without metabolic activation)
Positive controls:
yes
Positive control substance:
7,12-dimethylbenzanthracene
other: (with metabolic activation)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium

DURATION
- Exposure duration:
Experiment I: 4 h with and without metabolic activation
Experiment II: 4 h with metabolic activation, 24 h without metabolic activation
- Expression time (cells in growth medium): 7 days
- Selection time (if incubation with a selection agent): 8 days
- Fixation time (start of exposure up to fixation or harvest of cells): 15 days

SELECTION AGENT (mutation assays): 11 µg/mL 6-thioguanine (6-TG)

NUMBER OF REPLICATIONS: 2 replicates per experiment

DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency

OTHER:
- colonies used to determine the cloning efficiency (survival) were fixed and stained approx. 7 days after treatment
- the colonies were stained with 10 % methylene blue in 0.01 % KOH solution; colonies with more than 50 cells were counted.
- Following the expression time of 7 days five cell culture flasks were seeded with about 3 - 5E05 cells each in medium containing 6-TG; 2 additional flasks were seeded with approx. 500 cells each in non-selective medium to determine the viability.
Evaluation criteria:
The assay is considered acceptable if it meets the following criteria:
- the numbers of mutant colonies per 10E06 cells found in the solvent controls fall within the laboratory historical control data range
- the positive control substances must produce a significant increase in mutant colony frequencies
- the cloning efficiency (absolute value) of the solvent controls must exceed 50%

A positive response is described as follows:
- reproducible induction of a mutation frequency that is three times above the spontaneous mutation frequency at least at one of the concentrations in the experiment
- reproducible concentration-related increase of the mutation frequency; such evaluation may be considered also in the case that a threefold increase of the mutant frequency is not observed
Statistics:
A linear regression (least squares) was performed to assess a possible dose dependent increase of mutant frequencies. The number of mutant colonies obtained for the groups treated with the test item were compared to the solvent control groups. A trend is judged as significant whenever the p-value (probability value) is below 0.05.
Biological and statistical significance were considered together.
Species / strain:
Chinese hamster lung fibroblasts (V79)
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: Exp. I: no cytotoxicity with and without S9; Exp II: cytotoxic without S9, no cytotoxicity with S9
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: 7.4 for solvent control, 7.42 for test item -> no relevant influence
- Effects of osmolality: 383 mOsmfor solvent control, 374 mOsm for test item -> no relevant influence
- Precipitation: No precipitation of the test item in the culture medium was observed.

RANGE-FINDING/SCREENING STUDIES: pre-test for cytotoxicity, cytotoxic after 24 h exposure at 430.0 µg/mL and above without metabolic activation

COMPARISON WITH HISTORICAL CONTROL DATA: mutant frequencies were within the historical range of solvent controls, except the first culture of the first experiment without metabolic activation at 107.5 µg/mL, which was slightly above the historical range, however, this was neither reproduced nor dose dependent

ADDITIONAL INFORMATION ON CYTOTOXICITY: Relevant cytotoxic effects indicated by a relative cloning efficiency I or cell density below 50% occurred in experiment II at 430.0 µg/mL and above without metabolic activation.

No relevant and reproducible increase in mutant colony numbers/1E06 cells was observed in the main experiments up to the maximum concentration. The induction factor exceeded the threshold of three times the mutation frequency of the corresponding solvent control in the first culture of the first experiment without metabolic activation at 107.5 and 1720µg/mL. However, the increases were neither reproduced nor dose dependent as indicated by the lacking statistical significance. Consequently, the isolated increases of the mutation frequency were judged as biologically irrelevant and based on a relatively low solvent control.

 

Conc. [mg/L]

S9 mix

Rel. CE I (survival) [%]

Mutant colonies /1E06 cells

Induction factor

Rel. CE I (survival) [%]

Mutant colonies /10E06 cells

Induction factor

Experiment I, 4 h treatment

Culture I

Culture II

Solvent control

---

-

100.0

6.2

1.0

100.0

13.7

1.0

Positive control (EMS)

150.0

-

80.5

87.2

14.0

88.7

121.6

8.9

Test item

53.8

-

82.6

#

#

87.8

#

#

Test item

107.5

-

99.0

44.0

7.1

89.3

4.5

0.3

Test item

215.0

-

86.8

15.9

2.6

94.2

12.0

0.9

Test item

430.0

-

90.4

3.8

0.6

83.4

3.3

0.2

Test item

860.0

-

89.5

9.1

1.5

78.4

9.5

0.7

Test item

1720.0

-

88.8

24.1

3.9

84.6

7.7

0.6

Experiment I, 4 h treatment

Culture I

Culture II

Solvent control

---

+

100.0

6.5

1.0

100.0

8.2

1.0

Positive control (DMBA)

1.1

+

84.2

872.2

133.8

91.9

759.8

92.1

Test item

53.8

+

92.0

#

#

94.6

#

#

Test item

107.5

+

93.4

6.2

0.9

92.9

14.5

1.8

Test item

215.0

+

91.8

8.2

1.3

95.9

7.8

0.9

Test item

430.0

+

89.8

14.1

2.2

90.1

6.9

0.8

Test item

860.0

+

91.7

5.5

0.8

92.6

7.0

0.9

Test item

1720.0

+

93.3

6.0

0.9

91.9

5.5

0.7

Experiment I, 24 h treatment

Culture I

Culture II

Solvent control

---

-

100.0

13.7

1.0

100.0

23.1

1.0

Positive control (EMS)

150.0

-

95.3

746.0

54.5

95.5

540.5

23.3

Test item

107.5

-

102.8

27.5

2.0

105.9

11.8

0.5

Test item

215.0

-

103.5

25.6

1.9

90.4

8.8

0.4

Test item

430.0

-

35.6

10.5

0.8

36.1

9.7

0.4

Test item

860.0

-

35.8

11.4

0.8

39.8

15.6

0.7

Test item

1290.0

-

0.0

##

##

0.0

##

##

Test item

1720.0

-

0.0

##

##

0.0

##

##

Experiment I>I, 4 h treatment

Culture I

Culture II

Solvent control

---

+

100.0

27.3

1.0

100.0

8.9

1.0

Positive control (DMBA)

1.1

+

95.6

649.0

23.8

94.8

756.1

85.3

Test item

107.5

+

102.3

#

#

97.4

#

#

Test item

215.0

+

97.7

16.6

0.6

97.1

14.3

1.6

Test item

430.0

+

96.1

19.6

0.7

94.1

17.9

2.0

Test item

860.0

+

98.3

14.3

0.5

92.7

20.7

2.3

Test item

1290.0

+

98.3

18.7

0.7

93.3

6.4

0.7

Test item

1720.0

+

95.0

21.4

0.8

95.3

4.5

0.5

 

# culture was not continued since a minimum of only four analysable concentrations is required

## culture was not continued due to exceedingly severe cytotoxic effects

Conclusions:
In this HPRT test, Tetrahydrofurfuryl methacrylate was found to be not mutagenic.
Executive summary:

In a mammalian cell gene mutation assay according to OECD guideline 476, adopted 21 July 1997 (HPRT test) and EU Method B.17 dated May 30, 2008, V79 cells cultured in vitro were exposed toTetrahydrofurfuryl methacrylate (99.0% a.i.) at the following concentrations in the presence and absence of mammalian metabolic activation (S9 mix).

Experiment I (4 h exposure, with and without S9): 53.8, 107.5, 215.0, 430.0, 860.0, 1720.0 µg/mL

Experiment II (24 h exposure, without S9): 107.5, 215.0, 430.0, 860.0, 1290.0, 1720.0 µg/mL

Experiment II (4 h exposure, with S9): 107.5, 215.0, 430.0, 860.0, 1290.0, 1720.0 µg/mL

The test item was tested up to cytotoxic or the guideline limit concentration of 10 mM (corresponding to 1720.0 µg/mL). The positive controls induced the appropriate response.

No relevant and reproducible increase in mutant colony numbers/1E6 cells was observed in the main experiments up to the maximum concentration. The induction factor exceeded the threshold of three times the mutation frequency of the corresponding solvent control in the first culture of the first experiment without metabolic activation at 107.5 and 1720µg/mL. However, the increases were neither reproduced nor dose dependent as indicated by the lacking statistical significance. Consequently, the isolated increases of the mutation frequency were judged as biologically irrelevant and based on a relatively low solvent control.

There was no evidence of induced mutant colonies over background. In this HPRT test,Tetrahydrofurfuryl methacrylate was found to be not mutagenic.

NOTE: Any of data in this dataset are disseminated by the European Union on a right-to-know basis and this is not a publication in the same sense as a book or an article in a journal. The right of ownership in any part of this information is reserved by the data owner(s). The use of this information for any other, e.g. commercial purpose is strictly reserved to the data owners and those persons or legal entities having paid the respective access fee for the intended purpose.

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

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

For the assessment of the genotoxic potential of Tetrahydrofurfuryl methacrylate (THFMA), an analogue substance of the registered substance (THFA), the following reliable, relevant and adequate studies are available: a reverse gene mutation assay in bacteria, a mammalian cell gene mutation assay (HPRT) and a mammalian cell micronucleus assay.

 

In a reverse gene mutation assay in bacteria similar to OECD guideline 471, S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 strains were exposed to THFMA in DMSO at concentrations of 0, 4.88, 19.5, 78.1, 313, 1250, 5000 µg/plate in the first experiment and 0, 156, 313, 625, 1250, 2500, 5000 µg/plate in the second experiment in the presence and absence of mammalian metabolic activation (S9 mix).  THFMA was tested up to limit concentrations (5000 µg/plate).

Cytotoxic effects (reduced bacterial growth) were observed in strains TA98, TA100, TA1535 and TA1537 at 5000 µg/plate without metabolic activation and in strains TA100 and TA1535 at 5000 µg/plate with metabolic activation.

There was no evidence of induced mutant colonies over background. 

THFMA was not mutagenic in this bacterial reverse gene mutation assay when tested up to cytotoxic or limit concentrations (5000 µg/plate).

 

In a mammalian cell gene mutation assay according to OECD guideline 476, adopted 21 July 1997 (HPRT test) and EU Method B.17 dated May 30, 2008, V79 cells cultured in vitro were exposed to Tetrahydrofurfuryl methacrylate (99.0% a.i.) at the following concentrations in the presence and absence of mammalian metabolic activation (S9 mix).

Experiment I (4 h exposure, with and without S9): 53.8, 107.5, 215.0, 430.0, 860.0, 1720.0 µg/mL

Experiment II (24 h exposure, without S9): 107.5, 215.0, 430.0, 860.0, 1290.0, 1720.0 µg/mL

Experiment II (4 h exposure, with S9): 107.5, 215.0, 430.0, 860.0, 1290.0, 1720.0 µg/mL

The test item was tested up to cytotoxic or the guideline limit concentration of 10 mM (corresponding to 1720.0 µg/mL). The positive controls induced the appropriate response.

No relevant and reproducible increase in mutant colony numbers/1E6 cells was observed in the main experiments up to the maximum concentration. The induction factor exceeded the threshold of three times the mutation frequency of the corresponding solvent control in the first culture of the first experiment without metabolic activation at 107.5 and 1720µg/mL. However, the increases were neither reproduced nor dose dependent as indicated by the lacking statistical significance. Consequently, the isolated increases of the mutation frequency were judged as biologically irrelevant and based on a relatively low solvent control.

There was no evidence of induced mutant colonies over background. In this HPRT test, Tetrahydrofurfuryl methacrylate was found to be not mutagenic.  

In a mammalian cell micronucleus assay according to OECD guideline 487 (adopted July 22, 2010) and EU method B.49, dated July 06, 2012, primary human lymphocyte cultures were exposed to Tetrahydrofurfuryl methacrylate (99.0%) in DMSO with and without metabolic activation (S9 mix).

The following concentrations were evaluated (calculations were not adjusted to purity):

Experiment I:

4 h exposure, 16 h recovery, fixation time 40 h, without S9: 555.1, 971.4, 1700.0 µg/mL

4 h exposure, 16 h recovery, fixation time 40 h, with S9: 555.1, 971.4, 1700.0 µg/mL

Experiment II:

20 h exposure, fixation time 40 h, without S9: 317.2, 555.1, 971.4 µg/mL

4 h exposure, 16 h recovery, fixation time 40 h, with S9: 555.1, 971.4, 1700.0 µg/mL

The test item was tested up to cytotoxic or the guideline limit concentration of 10 mM (corresponding to 1700.0 µg/mL). No cytotoxicity was observed in Experiment I with and without metabolic activation as well as in Experiment II with metabolic activation. In Experiment II in the absence of S9 mix the highest applied concentration showing clear cytotoxic effects was not evaluable.

In both independent experiments, neither a statistically significant nor a biologically relevant increase in the number of micronucleated cells was observed after treatment with the test item. 

Positive controls induced the appropriate response. 

There was no evidence of micronucleated cells induced over background. Therefore, Tetrahydrofurfuryl methacrylate is considered to be non-clastogenic in this in vitro micronucleus test, when tested up to cytotoxic or the guideline limit concentration.

 

There are no data gaps for the endpoint genotoxicity. No human information is available for this endpoint. However, there is no reason to believe that these results would not be applicable to humans. 

Justification for classification or non-classification

Based on the available data, the registered substance (THFA) does not need to be classified for mutagenicity according to the criteria given in regulation (EC) 1272/2008