Hydrogen trifluoromethoxyborate(1-), compound with methanol (1:1)

Genetic toxicity: in vivo

Administrative data

Endpoint:

genetic toxicity in vivo, other

Remarks:

cytogenicity

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH


1. HYPOTHESIS FOR THE ANALOGUE APPROACH
Similar to all coordination complexes of boron trifluoride with organic and inorganic species (like alcohols, ethers, amines, sulfuric acid, sulfuric dioxide, etc) the complex of boron trifluoride and methanol is extremely water sensitive and reacts even with moist air. Given the extremely rapid decomposition the rate of hydrolysis cannot be quantitatively derived in a study conducted according to OECD Guideline 111. However sufficient information is available on the identity of hydrolysis products which aquatic organisms may be eposed.
In the instantaneous reaction with water as a first step methanol and boron trifluoride dihydrates are formed.
BF3·CH3OH+ 2 H2O -> BF3· 2 H2O + CH3OH
The hydrolysis of borontrifluoride dihydrate was investigated at pH-values of 1.2; 4.0; 7.0 and 9.0 (BASF SE, Study No. 09S01179, 26.19.2009)
The hydrolysis of the test item is a very fast process (half-life < 30 minutes). Already at room temperature about 30 to 60% of the test item was hydrolyzed within minutes in all tested buffer solutions in the pH range 1.2 to 9.0.
The main reactions of the hydrolysis process are the following:
BF3.2H2O + H2O → B(OH)3+ 3HF
HF + BF3.2H2O → HBF4+ 2H2O
B(OH)3+ 4HF → HBF4+ 3 H2O
HBF4+ H2O ↔ HBF3(OH) + HF
Total hydrolysis of HBF4:
HBF4↔ HBF3(OH) ↔ HBF2(OH)2↔ HBF(OH)3↔ HB(OH)4= B(OH)3+ H2O
Boric acid and tetrafluoroborate are expected to be the main species in the dilute hydrolysis solution (Zhang Weijiang et al.).
In conclusion it is justified to base the assessment of environmental fate and pathways of hydrogen trifluoromethoxyborate (1-), compound with methanol (1.1) on the properties of the breakdown products, which is considered acceptable in accordance with REACH Annex XI, section 1.5.
References:
BASF SE, Boron trifluoride dihydrate: hydrolysis as a function of pH, Study No. 09S01179, 26.10.2009
Wamser C. A. Equilibria in the system boron trifluoride-water at 25 °C,Journal of the American Chemical Society, 1951, 73: 409-416
Zhang Weijiang et al., Equilibrium on the hydrolysis of Boron trifluoride in large amount of water,Transactions of Tianjin University, 2016, 22: 486-491


2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
methanol (67-56-1). For purity and impurities see study records in IUCLID chapter 7.6.2.

3. ANALOGUE APPROACH JUSTIFICATION
The hydrolysis data in IUCLID chapter 5.1.2 demonstrate the rapidity of the hydrolysis reaction. Exposure will never occur to the complex of boron trifluoride methanol but to its hydrolysis products, BF3 dihydrate and methanol. Therefore, these two substances are the relevant ones for evaluation of potential health effects.

4. DATA MATRIX
A data matrix is not applicable in this case. The present read-across approach is not based on the comparison of experimental data for one or more compounds, but the hydrolysis products of the registered material are considered the relevant toxophors due to the rapid hydrolysis reaction. Reliable data are provided for all hydrolysis products, whereby an evaluation of potential health hazards can be performed.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

read-across source

Reference

Endpoint:

in vivo mammalian germ cell study: cytogenicity / chromosome aberration

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Test procedure based on scientific principles and standards, sufficiently documented, acceptable for assessment, however missing positive control.

Reason / purpose for cross-reference:

reference to same study

Principles of method if other than guideline:

Spermatocytes of the pachytene substage of the meiotic prophase after 5 days of last exposure were isolated and morphological parameters concerning the meiotic chromosome structure were examined.

GLP compliance:

not specified

Type of assay:

chromosome aberration assay

Species:

mouse

Strain:

other: C57BL/6J

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories
- Age at study initiation: 10 weeks old
- Housing: Mice were housed in an animal facility in laminar-flow rooms
- Diet (e.g. ad libitum): Purina rodent chow
- Water (e.g. ad libitum): tap water


ENVIRONMENTAL CONDITIONS
- Air changes (per hr): 15 cycles/hour of biocleaned air
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

inhalation: vapour

Vehicle:

none

Details on exposure:

TYPE OF INHALATION EXPOSURE: whole body


GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure: Methanol was vaporised and transported by nitrogen and HEPA-filtered air to stainless steel chambers.
- Air flow rate: 105 L/min
- Air change rate: 15 air changes/hours


TEST ATMOSPHERE
- Brief description of analytical method used:
Chamber methanol concentrations, monitored continuously by a Foxboro Miran 1A Infrared Analyser, indicated that ppm levels did not differ by more than 7% from calculated values.

Duration of treatment / exposure:

5 days

Frequency of treatment:

6 h/day

Post exposure period:

5 days

Remarks:

Doses / Concentrations:
1.04 or 5.3 mg/L (corresponding to 800 or 4000 ppm)
Basis:
nominal conc.

No. of animals per sex per dose:

5

Control animals:

yes

Positive control(s):

No positive control group was concomitantly examined.

Tissues and cell types examined:

Spermatocytes of the pachytene substage of the meiotic prophase 5 days after last exposure

Details of tissue and slide preparation:

Spermatocytes of the pachytene substage of the meiotic prophase after 5 days of last exposure were isolated, and fixed for electronmicroscopy: Several morphological parameters concerning the meiotic chromosome structure were examined (synaptonemal complex analysis).

Statistics:

A 1- way analysis of variance was performed with Statgraphics statistical package. SC data were then analysed by a 1-tailed Dunnett´s test.

Sex:

male

Genotoxicity:

negative

Remarks:

No increased frequencies of synaptonemal complex damage in spermatocytes were found.

Toxicity:

not specified

Vehicle controls validity:

not applicable

Negative controls validity:

not examined

Positive controls validity:

not examined

Additional information on results:

Methanol did not produce significant dose-dependent increases in SC aberrations.

Conclusions:

Interpretation of results (migrated information): negative

Cross-reference 2

Reason / purpose for cross-reference:

read-across source

Reference

Endpoint:

in vivo mammalian germ cell study: cytogenicity / chromosome aberration

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study and current standards, sufficiently documented, acceptable for assessment, however missing positive control.

Reason / purpose for cross-reference:

reference to same study

Principles of method if other than guideline:

Mammalian chromosome aberration test performed with primary lung cells cultured after inhalation exposure.

GLP compliance:

not specified

Type of assay:

chromosome aberration assay

Species:

mouse

Strain:

other: C57BL/6J

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories
- Age at study initiation: 10 weeks old
- Housing: Mice were housed in an animal facility in laminar-flow rooms
- Diet (e.g. ad libitum): Purina rodent chow
- Water (e.g. ad libitum): tap water


ENVIRONMENTAL CONDITIONS
- Air changes (per hr): 15 cycles/hour of biocleaned air
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

inhalation: vapour

Vehicle:

none

Details on exposure:

TYPE OF INHALATION EXPOSURE: whole body


GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure: Methanol was vaporised and transported by nitrogen and HEPA-filtered air to stainless steel chambers.
- Air flow rate: 105 L/min
- Air change rate: 15 air changes/hours


TEST ATMOSPHERE
- Brief description of analytical method used:
Chamber methanol concentrations, monitored continuously by a Foxboro Miran 1A Infrared Analyser, indicated that ppm levels did not differ by more than 7% from calculated values.

Duration of treatment / exposure:

5 days

Frequency of treatment:

6 h/day

Post exposure period:

no

Remarks:

Doses / Concentrations:
1.04 or 5.3 mg/L (corresponding to 800 or 4000 ppm)
Basis:
nominal conc.

No. of animals per sex per dose:

5

Control animals:

yes

Positive control(s):

No positive control group was concomitantly examined. Historical positive control are available.

Tissues and cell types examined:

primary cultures of lung cells

Details of tissue and slide preparation:

Immediately following the last exposure, animals were anesthetised and blood was removed by perfusion, the lungs were infused with a trypsin, EDTA and collagenase solution; and then removed, minced and incubated in the same enzyme solution. The cells were collected and culture dishes with 160,000 viable cells per animal were established. One hundred first-division metaphases per animal were examined for CA. Aberrations were classified separately by type (chromatid or chromosome) and analysed as rearrangement or breakage events. The replication indices were estimated from 200 metaphases per animal. The number of metaphases per 1000 consecutive cells was also counted for determination of mitotic indices.

Statistics:

A 1- way analysis of variance was performed with Statgraphics statistical package. CA data were then analysed by a 1-tailed Dunnett´s test.

Sex:

male

Genotoxicity:

negative

Remarks:

There was no evidence of induced chromosome aberrations in lung cells.

Toxicity:

no effects

Vehicle controls validity:

not applicable

Negative controls validity:

not examined

Positive controls validity:

other: only historical positive controls

Additional information on results:

No increase in the frequency of aberrations per cell and percent aberrant cells and no toxic effects were induced by the treatment:
Aberration rate per cell (including chromatid and chromosome breakage as well as rearrangement events) was from 0.04 to
0.07 in treated animals vs. 0.09 in the control group. The few aberrations found were mostly chromatid-type breaks or fragments.
Percentages of cells with abnormal chromosomes were not significantly different among treated and control groups within an experiment (6.9 % vs. 6.2 % (for 4000 ppm) and 3.8 % vs. 3.8 % (800 ppm). The replicative and mitotic indices were not significantly different from the controls.

Conclusions:

Interpretation of results (migrated information): negative

Cross-reference 3

Reason / purpose for cross-reference:

read-across source

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study and current standards, sufficiently documented, acceptable for assessment.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

yes

Remarks:

- limited documentation

GLP compliance:

yes

Type of assay:

micronucleus assay

Species:

mouse

Strain:

Swiss Webster

Sex:

male

Details on test animals or test system and environmental conditions:

no data

Route of administration:

intraperitoneal

Vehicle:

no data

Details on exposure:

no data

Duration of treatment / exposure:

4 days

Frequency of treatment:

no data

Post exposure period:

24 h

Remarks:

Doses / Concentrations:
300, 600, 1200, 2500 mg/kg bw
Basis:
nominal conc.

No. of animals per sex per dose:

10

Control animals:

yes

Positive control(s):

Caffeine (-folate)
Urethane (+folate)

Tissues and cell types examined:

erythrocytes from blood samples

Details of tissue and slide preparation:

see any other information on materials and methods

Evaluation criteria:

no data

Statistics:

no data

Sex:

male

Genotoxicity:

negative

Toxicity:

not specified

Vehicle controls validity:

not specified

Negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

see remarks on results

Folate limitation resulted in an about 15-20 times lower folate blood level than in the high-folate groups.

4/10 folate-deficient animals receiving 2500 mg/kg died between days 2 and 3.

No difference in micronucleus frequency (MN in PCEs) and in RNA-positive blood erythrocytes was seen between treated groups and controls while animals treated with the positive control substances responded as expected:

+folate: MN 0.17 - 0.23 % vs. 0.23 % in saline control

-folate: MN 0.31 - 0.37 % Vs. 0.38 % in saline control.

Caffeine (+folate): MN 0.34 %

Caffeine (-folate): MN 2.42 %

Urethane (+folate): MN 2.52 %.

The results indicate that methanol is nonclastogenic to the developing erythroblast in bone marrow and does not inhibit red blood cell production in either normal or folate-deficient mice.

Conclusions:

Interpretation of results (migrated information): negative

Cross-reference 4

Reason / purpose for cross-reference:

read-across source

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Test procedures in accordance with accepted standard methods, sufficiently documented.

Principles of method if other than guideline:

The study was performed during a developmental investigation under folate-deficient and -sufficient conditions.
After oral methanol application, the frequency of MN was examined in maternal peripheral blood and fetal blood.

GLP compliance:

not specified

Type of assay:

micronucleus assay

Species:

mouse

Strain:

CD-1

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Inc.
- Weight at study initiation: 12-14 g
- Housing: Weanling mice were housed in stainless steel, wire-bottomed cages.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-23 °C
- Humidity (%): 50 %
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

deionised water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
2.5 g/kg (15.65 % Optima HPLC grade MeOH in deionised water)

Duration of treatment / exposure:

gestation day 6 through 10 during pregnancy

Frequency of treatment:

twice daily

Post exposure period:

48 h after the final exposure: maternal peripheral blood was collected
gestation day 18: fetal blood was taken

Remarks:

Doses / Concentrations:
2500 mg/kg/d
Basis:
nominal conc.

No. of animals per sex per dose:

1 to 17 dam(s)

Control animals:

yes, concurrent vehicle

Positive control(s):

none

Tissues and cell types examined:

maternal peripheral and fetal blood

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION:
The dose of methanol was based on previous work [Sakanashi et al., 1994, Teratology].

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields):
Maternal vein blood (2 to 3 µL) was collected on precleaned microscope slides 48 hours (still during pregnancy) after the final methanol treatment.
Fetal blood was taken on gestation day 18.

DETAILS OF SLIDE PREPARATION:
Slides were air dried and fixed in absolute methanol on the same day. Fixed slides were stained with 0.1 % acridine orange for 5 min, followed by a 17 min rinse in Sorensen´s M/15 phosphate buffer, pH 6.8. Slides were then wet mounted with cover slips and examined by fluorescent microscopy using a mercury lamp.

METHOD OF ANALYSIS:
Micronuclei fluoresce green-yellow. The frequency of MN was scored in 1000 reticulocytes.

Statistics:

For all statistical the pregnant dams and their litters were considered the units for comparison. Continuous variables were analysed using the two-way analysis of variance procedure and the Fisher PLSD for multiple comparisons of means. These analyses were carried out on STATVIEW SE+ (Abacus, Berkeley, CA). Incidences of frequency of micronuclei, bases on affected litters, were analyses using binomial statistics.

Sex:

female

Genotoxicity:

negative

Remarks:

The frequency of micronuclei in maternal and fetal reticulocytes was not influenced by either the marginal folic acid diet (400 nmol/kg) or by methanol treatment.

Toxicity:

not specified

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

not examined

Additional information on results:

The frequency of MN in maternal and fetal reticulocytes was similar among the groups. The MN rate was not associated with either maternal or fetal hepatic folate concentrations. There was no significant difference in the frequency of MN in reticulocytes of fetuses that were affected by malformations compared to fetuses without malformations.

Conclusions:

Interpretation of results (migrated information): negative

Cross-reference 5

Reason / purpose for cross-reference:

read-across source

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study and current standards, sufficiently documented, acceptable for assessment, however missing positive control.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

yes

Remarks:

- primary lung cells in addition to erythrocytes, not included in the guideline; no positive control (only historical)

GLP compliance:

not specified

Type of assay:

micronucleus assay

Species:

mouse

Strain:

other: C57BL/6J

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories
- Age at study initiation: 10 weeks old
- Housing: Mice were housed in an animal facility in laminar-flow rooms
- Diet (e.g. ad libitum): Purina rodent chow
- Water (e.g. ad libitum): tap water


ENVIRONMENTAL CONDITIONS
- Air changes (per hr): 15 cycles/hour of biocleaned air
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

inhalation: vapour

Vehicle:

none

Details on exposure:

TYPE OF INHALATION EXPOSURE: whole body


GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure: Methanol was vaporised and transported by nitrogen and HEPA-filtered air to stainless steel chambers.
- Air flow rate: 105 L/min
- Air change rate: 15 air changes/hours


TEST ATMOSPHERE
- Brief description of analytical method used:
Chamber methanol concentrations, monitored continuously by a Foxboro Miran 1A Infrared Analyser, indicated that ppm levels did not differ by more than 7% from calculated values.

Duration of treatment / exposure:

5 days

Frequency of treatment:

6 h/day

Post exposure period:

no

Remarks:

Doses / Concentrations:
1.04 or 5.3 mg/L (corresponding to 800 or 4000 ppm)
Basis:
nominal conc.

No. of animals per sex per dose:

5

Control animals:

yes

Positive control(s):

No positive control group was concomitantly examined. Historical positive controls are available.

Tissues and cell types examined:

peripheral blood; primary cultures of lung cells

Details of tissue and slide preparation:

Peripheral blood cell MN analysis:
Immediately following the last exposure, animals were anesthetised and blood smears were made from tail vein blood for erythrocyte MN determination. The cells were fixed in methanol and stained with acridine orange for fluorescence microscopy. All slides were coded prior to scoring of 2000 polychromatic erythrocytes (PCE) and 2000 normochromatic erythrocytes (NCE) per animal.

Lung cell MN analysis:
The same exposure groups of mice used for blood MN analysis were also used for lung cell MN analysis. After blood was removed by perfusion, the lungs were infused with a trypsin, EDTA and collagenase solution; and then removed, minced and incubated in the same enzyme solution. The cells were collected and culture dishes with 160,000 viable cells per animal were established. Lung MN were analysed in 1000 binucleated cells typically examined from each of 5 animals per dose. Percentages of mononucleated, binucleated, trinucleated and quadrinucleatexd cells were also determined.

Statistics:

A 1- way analysis of variance was performed with Statgraphics statistical package. MN data were then analysed by a 1-tailed Dunnett´s test.

Sex:

male

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

not applicable

Negative controls validity:

not examined

Positive controls validity:

other: historical positive controls

Additional information on results:

No treatment related effect on micronuclei frequency or cell kinetics and no toxicity were seen at any dose level:

Peripheral blood cell MN analysis:
Mean MN rates were 3.1/1000 and 5.2/1000 in PCE, and 3.4/1000 and 3.6/1000 in NCE at either dose, respectively, vs. 5.0/1000 and 3.7/1000 of respective controls.

Lung cell MN analysis:
Mean MN rates were about 20/1000 to 24/1000 irrespective of a dose or control group. The ratio of bi- to mononucleated cells was not influenced by the treatment, neither was there evidence of a treatment-related increase in the incidence of multi-nucleated cells.

Conclusions:

Interpretation of results (migrated information): negative

Cross-reference 6

Reason / purpose for cross-reference:

read-across: supporting information

Reference

Endpoint:

in vivo mammalian cell study: DNA damage and/or repair

Remarks:

Type of genotoxicity: DNA damage and/or repair

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: scientifically acceptable publication

Principles of method if other than guideline:

This study was performed to determine whether MeOH could indirectly DNA damage via ROS-mediated mechanism. Animals (mice, rabbits and monkeys) were treated with either once or for 15 days with 2.0 g/kg MeOH i.p.. Oxidative DNA damage was observed by measuring 8-oxo-2'-deoxyguanosine. Lipid peroxidation in bone marrow and spleen homogenates was determined by measuring the Ievels of HNE-His protein adducts.

GLP compliance:

not specified

Type of assay:

other: oxidative DNA damage

Species:

other: mouse, rabbit, monkey

Strain:

other: CD-1, New Zealand white, cynomolgus

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: mice, rabbits and monkeys: Charles River Laboratories
- Age at study initiation: mice: 9 - 13 weeks; rabbits: 5 months; monkeys: 3.4 - 5.7 years
- Weight at study initiation: rabbits: 3.25 - 3.75 kg, monkeys: 2.8 - 4.8 kg
- Diet: mice: rodent chow ad libitum; rabbit: standard high-fiber rabbit chow; monkeys: certified primate chow diet (# 5048) from Purina Mills (St. Louis, MO) supplemented with fruit or vegetables 2-3 times weekly
- Water: mice, rabbit and monkeys: ad libitum
- Acclimation period: monkeys: two weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): mice and rabbits: 20°C; monkeys: 18 - 29°C
- Humidity (%): mice: 50%, rabbits: 60%
- Photoperiod (hrs dark / hrs light): mice: 10/14; rabbits and monkeys: 12/12

Route of administration:

intraperitoneal

Vehicle:

- Vehicle(s)/solvent(s) used: saline

Details on exposure:

Mice: Drugs were administered via intraperitoneal (ip) injection using a 26 gauge (G) 3/8 needle.
Rabbits: by ip injection usinga 23 G needle
monkeys: were lightly sedated with ketamine (ca. 5 - 10 mg/kg) for dose administration and then administered MeOH (2g/kg bw; 20% [w/v] in sterile saline) or a saline vehicle by ip injection using a 22G needle.

Duration of treatment / exposure:

single or 15 consecutive days

Frequency of treatment:

daily

Post exposure period:

6 hours or 15 days

Remarks:

Doses / Concentrations:
2 g/kg bw
Basis:
nominal conc.

No. of animals per sex per dose:

Three rabbits and primates were used in each treatment group. Four and five mice were used in each group for the acute and chronic studies, respectively.

Control animals:

yes, concurrent vehicle

Positive control(s):

KBrO3
- Justification for choice of positive control(s): is a known renal carcinogen
- Route of administration: i.p.
- Doses / concentrations: 100 mg/kg bw at a fixed volume of 0.1 mL/10g bw

Tissues and cell types examined:

DNA from spleen and bone marrow

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: The effect of a Iimit dose of 2.0 g/kg bw MeOH based on guidelines established for the comet assay developed in accordance with the in vivo genetic toxicology guidelines of the Organization for Econornic Co-operation and Development (OECD) were studied.

METHOD OF ANALYSIS: DNA was isolated using DNAzol, a novel guanidine-detergent lysing solution that hydrolyzes RNA and allows for the selective precipitation of DNA from cell lysates. 8-oxodG and dG were analysed via HPLC system. Lipid peroxidation in bone marrow and spleen homogenates was determined by measuring the Ievels of HNE-His protein adducts.

Statistics:

Statistical analysis was performed using GraphPad Instat Version 3.05 (GraphPad Software, lnc., San Diego, CA). Experiments comparing two groups were analyzed by unpaired t tests and multiple comparisons were anaJyzed by one-way ANOVA followed by a Tukey post-test. The Ievel of significance was set at P<0.05.

Sex:

male

Genotoxicity:

negative

Toxicity:

not examined

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

No increase in oxidative DNA damage was observed in bone marrow or spleen at 6 h following exposure to MeOH (2.0g/kg bw ip) in mice, rabbits,
or primates. Similarly, no increase in oxidative DNA damage was observed in bone marrow or spleen 24 h following exposure to a single dose of MeOH (2.0g/kg bw ip), or following 15 consecutive daily doses of 2.0g/kg bw ip MeOH in CD-1 mice. No increase in HNE-His protein adducts was observed in bone marrow or spleen at 6 h following exposure to MeOH (2.0 g/kg bw ip) in primates. MeOH exposure (2.0 g/kg bw ip) increased HNE-His protein adducts 1.4-fold 6 h post-dose in bone marrow of mice, with adduct Ievels returning to basal values within 24 h. No increases in HNE-His protein adducts were observed in spleen of mice or bone marrow of rabbits following MeOH exposure (2.0 g/kg bw ip). MeOH exposure (2.0 g/kg bw ip) increased HNE-His protein adducts 1.5-fold 6 h post-dose in spleen of rabbits.

Conclusions:

Interpretation of results (migrated information): negative
Taken together these observations suggest that it is unlikely that exposure to MeOH would initiate Iymphomas, particularly in humans, via formation
of mutagenic oxidative DNA lesions.

Cross-reference 7

Reason / purpose for cross-reference:

read-across: supporting information

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study and current standards, limited documentation, restriction: only one sampling time

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

yes

Remarks:

- limited documentation, only one sampling time

GLP compliance:

not specified

Type of assay:

micronucleus assay

Species:

mouse

Strain:

NMRI

Sex:

male/female

Route of administration:

intraperitoneal

Duration of treatment / exposure:

one single injection

Frequency of treatment:

only one sampling time

Post exposure period:

30 h

Remarks:

Doses / Concentrations:
1920; 3200; 4480 mg/kg
Basis:
nominal conc.

No. of animals per sex per dose:

2

Control animals:

yes

Tissues and cell types examined:

bone-marrow cells

Sex:

male/female

Genotoxicity:

negative

Toxicity:

not specified

Vehicle controls validity:

not applicable

Negative controls validity:

not examined

Positive controls validity:

not examined

There was no significant effect on the frequency of micronuclei in treated as compared to control animals: 1.3/1000 (control and low dose), 2.0/1000 (2nd dose), and 3.7/1000 (top dose).

Conclusions:

Interpretation of results (migrated information): negative

Materials and methods

Test material

Test material information

Constituent 1

Reference substance name:

Hydrogen trifluoromethoxyborate(1-), compound with methanol (1:1)

EC Number:

220-543-9

EC Name:

Hydrogen trifluoromethoxyborate(1-), compound with methanol (1:1)

Cas Number:

2802-68-8

Molecular formula:

CH4O.CH3BF3O.H

IUPAC Name:

hydrogen trifluoro(methanolato)borate(1-) methanol (1:1)

Results and discussion

Test results

Test results 1

Sex:

male

Genotoxicity:

negative

Toxicity:

not specified

Vehicle controls validity:

not specified

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

other: American Petroleum Institute, 1991

Test results 2

Sex:

female

Genotoxicity:

negative

Toxicity:

not specified

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

not examined

Remarks on result:

other: Fu et al., 1996

Test results 3

Sex:

male

Genotoxicity:

negative

Toxicity:

not specified

Vehicle controls validity:

not applicable

Negative controls validity:

not examined

Positive controls validity:

not examined

Remarks on result:

other: Campell et al., synaptonemal complex, 1991

Test results 4

Sex:

male

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

not applicable

Negative controls validity:

not examined

Positive controls validity:

other: historical positive controls

Remarks on result:

other: Campell et al., MN, 1991

Test results 5

Sex:

male

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

not applicable

Negative controls validity:

not examined

Positive controls validity:

other: historical positive controls

Remarks on result:

other: Campell et al., CA lung cells mouse, 1991

Test results 6

Sex:

male/female

Genotoxicity:

negative

Toxicity:

not specified

Vehicle controls validity:

not applicable

Negative controls validity:

not examined

Positive controls validity:

not examined

Remarks on result:

other: Gocke et al., 1981

Test results 7

Sex:

female

Genotoxicity:

negative

Toxicity:

not examined

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

other: McCallum et al., 2010