Registration Dossier
Registration Dossier
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EC number: 248-704-9 | CAS number: 27871-49-4
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
No target substance specific data is available for assessing the genotoxic potential of Methyl (S)-lactate. Thus, suitable data from read-across substances were used in a weight-of-evidence approach.
L-lactic acid was tested negative in an in vitro genotoxicity test battery (OECD TG 471, 473 & 476 (nowadays 490). Ethyl lactate was tested negative in a bacterial reverse mutation assay. Ethyl (S)-lactate was tested negative in two studies conducted according to OECD TG 473 and OECD TG 476 (nowadays OECD 490). Methanol was tested negative in an in vitro mammalian cell micronucleus test and in a bacterial reverse mutation assay (Ames test). Based on the available data from suitable read-across partners, the target substance can be considered to be non-genotoxic.
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- For details and justification of read-across please refer to the report attached in section 13 of IUCLID.
- Reason / purpose for cross-reference:
- read-across source
- Species / strain:
- S. typhimurium TA 1538
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Remarks:
- see Table 2
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Remarks:
- see Table 2
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Remarks:
- see Table 2
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Remarks:
- see Table 2
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Remarks:
- see Table 2
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- RANGE-FINDING/SCREENING STUDIES:
The results of the dose range-finding study conducted in the presence and absence of rat liver microsomes indicate that no appreciable toxicity was observed up to 10000 µg per plate. - Conclusions:
- Under the experimental conditions reported, Ethyl lactate did not cause gene mutations in an Ames Test (equivalent to OECD Guideline 471). Therefore, the test item is considered to be non-mutagenic in this bacterial reverse gene mutation assay.
- Executive summary:
In a reverse gene mutation assay (equivalent to OECD Guideline 471) in bacteria, strains TA98, TA100, TA1535, TA1537 and TA1538 of S. typhimurium were exposed to Ethyl lactate at concentrations of 667, 1000, 3333, 6667 and 10000 µg/plate in the presence and absence of mammalian metabolic activation (plate co-incubation).
Ethyl lactate was tested up to the limit concentration of 10000 µg/plate. There was no dose-related increase in the number of revertants in any of the test strains with and without activation. The positive controls induced the appropriate responses in the corresponding strains. There was no evidence of induced mutant colonies over background.
This study is classified as acceptable. This study satisfies the requirement for OECD test guideline 471 for in vitro mutagenicity (bacterial reverse gene mutation) data.
This information is used in a read-across approach in the assessment of the target substance. For justification of read-across please refer to the attached read-across report (see IUCLID section 13).
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- For details and justification of read-across please refer to the report attached in section 13 of IUCLID.
- Reason / purpose for cross-reference:
- read-across source
- Key result
- Species / strain:
- lymphocytes: peripheral human lymphocytes
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Conclusions:
- Ethyl (S)-lactate is considered to be not clastogenic in the in vitro mammalian chromosomal aberration test using human lymphocytes, with and without metabolic activation.
- Executive summary:
In a mammalian cell cytogenetics assay (chromosome aberration) according to OECD Guideline 473, primary peripheral human lymphocytes cultures were exposed to Ethyl (S)-lactate (purity 99.83%) at concentrations of 0, 100, 333 and 1180 µg/mL with and without metabolic activation.
The test item was tested up to the limit concentration of 1180 µg/mL (0.01 M). Positive controls induced the appropriate response. There was no evidence of chromosome aberration induced over background.
In conclusion, Ethyl (S)-lactate is not clastogenic in human lymphocytes.
This study is classified as acceptable and satisfies the requirement for the in vitro mammalian chromosomal aberration test according to OECD 473.
For details and justification of read-across please refer to the report attached in section 13 of IUCLID.
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- For details and justification of read-across please refer to the read-across report attached to IUCLID section 13.
- Reason / purpose for cross-reference:
- read-across source
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Species / strain:
- S. typhimurium TA 1538
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Conclusions:
- Based on the results, the test item showed no evidence of mutagenic activity in a bacterial reverse mutation assay.
- Executive summary:
The plate-incorporation test was performed according to the standard procedure as described by Ames et al., 1975 (similar/equivalent to OECD TG 471). Methanol was assayed in the Ames test with the S. typhimurium strains TA1535, TA1537, TA1538, TA98 and TA100 both with and without S9 mix at a concentration of up to 2.5 x 10^6 nmol/plate. Based on the results, there was no evidence of induced mutant colonies in comparison to controls.
This information is used in a read-across approach in the assessment of the target substance. For details and justification of read-across please refer to the read-across report attached to IUCLID section 13
- Endpoint:
- in vitro cytogenicity / micronucleus study
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- For details and justification of read-across please refer to the read-across report attached to IUCLID section 13.
- Reason / purpose for cross-reference:
- read-across source
- Details on test system and experimental conditions:
- METHOD OF TREATMENT/ EXPOSURE:
- Cell density at seeding : 13000 cells/cm²
- Test substance added in medium
TREATMENT AND HARVEST SCHEDULE:
- Preincubation period: 15-18 hours
- Exposure duration/duration of treatment: 1 hour
- Harvest time after the end of treatment : 48 hours
After treatment, the cells were incubated for 48 hours, then collected, subjected to hypotonic treatment with KCl, fixed with acetic acid-methanol, and stained with 4% Giemsa. A 7000 interphase cells with visible cytoplasm were scored per treatment.
The results are expressed as the mean number of cells with MN per 1000 cells ± SE - Species / strain:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not specified
- Positive controls validity:
- valid
- Conclusions:
- In conclusion, it can be stated that methanol did not induce structural and/or numerical chromosomal damage in Chinese hamster V79 cells. Therefore, the test substance is considered to be non-mutagenic with respect to clastogenicity and/or aneugenicity in this in vitro mammalian cell micronucleus test.
- Executive summary:
In an in vitro mammalian cell micronucleus assay conducted similar to OECD test guideline 487, V79 Chinese hamster cells cultured in vitro, were exposed for 1 hour to methanol at a concentration of 50 µL/mL without metabolic activation.
No increase of the micronucleus frequency was noted after treatment with the test item. The positive controls did induce distinct and biologically relevant increases of the micronucleus frequency. Therefore, it can be stated that methanol is considered to be non-mutagenic under the described test conditions.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- For details and justification of read-across please refer to the read-across report attached to IUCLID section 13.
- Reason / purpose for cross-reference:
- read-across source
- Key result
- Species / strain:
- lymphocytes: peripheral human lymphocytes
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- based on determination of the mitotic index.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: At the highest concentration of the test substance (10 mM equal to 901 µg/ml) the pH was 7.1 compared to a pH of 7.8 in the solvent control.
- Effects of osmolality: At the highest concentration of the test substance (10 mM equal to 901 µg/mL) the osmolarity was 275 mOsm/kg compared to an osmolarity of 269 mOsm/kg in the solvent control.
- Water solubility: miscible
- Precipitation: No
RANGE-FINDING/SCREENING STUDIES: In the dose range finding test blood cultures were treated with 10, 33, 100, 333, 901 µg/mL L(+)-lactic acid/mL culture medium (equal to concentrations of 0.1, 0.4, 1.1, 3.7 and 10 mM) with and without S9-mix.
COMPARISON WITH HISTORICAL CONTROL DATA: The number of cells with chromosome aberrations found in the solvent control cultures was within the laboratory historical control data range.
For individual results see box 'Any other information on results incl. tables'. - Conclusions:
- L(+)-lactic acid is considered to be not clastogenic in the in vitro mammalian chromosomal aberration test conducted according to the OECD TG 473 using human lymphocytes in the presence or absence of metabolic activation.
- Executive summary:
In an in vitro cytogenicity assay conducted according to OECD TG 473, peripheral human lymphocyte cultures were exposed to L(+)-lactic acid (90% purity), solved in RPMI 1640 cell culture medium. In the first experiment, the doses were 0, 10, 100, 901 µg/mL with and without metabolic activation. In the second experiment doses were 0, 100, 333, 666, 901 µg/mL without metabolic activation and 0, 10, 100, 901 µg/mL with metabolic activation (rat liver S9-mix).
L(+)-lactic acid was tested up to 901 µg/mL, which was cytotoxic based on determination of the mitotic index after an exposure time of 24 and 48 hours. The percentage of the mitotic index after 24 hours of 901 µg/mL was 55%, that after 48 hours of 901 µg/mL 49%. Concentrations lower than 901 µg/mL did not cause a dose-dependent decrease in the percentage of the mitotic index after 24 and 48 hours of exposure. The mitotic index after 3 hours of exposure was lower compared to control (66% in experiment 1, 84% in experiment 2) but did not reach the threshold value of 45 ± 5% according to OECD guideline 473 for cytotoxicity. Positive controls induced the appropriate responses. There was no evidence for a concentration related positive response of chromosome aberration induced over background.
This study is classified as acceptable and satisfies the requirement for the in vitro mammalian chromosomal aberration test according to OECD 473.
This information is used in a read-across approach in the assessment of the target substance. For details and justification of read-across please refer to the read-across report attached to IUCLID section 13.
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- For details and justification of read-across please refer to the read-across report attached to IUCLID section 13.
- Reason / purpose for cross-reference:
- read-across source
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: precipitation of the test material was not observed in any plate of the tester strains.
RANGE-FINDING/SCREENING STUDIES:
- Precipitation: precipitation of the test material was not observed in any plate of the tester strains.
- Toxicity: no reduction of the bacterial background lawn and no biologically relevant decrease in the number of revertants were observed.
ADDITIONAL INFORMATION ON CYTOTOXICITY: no reduction of the bacterial background lawn and no biologically relevant decrease in the number of revertants were observed. - Conclusions:
- Propyl (S)-lactate is not genotoxic in the bacterial reverse gene mutation assay (OECD 471) in the presence and absence of mammalian metabolic activation.
- Executive summary:
In a reverse gene mutation assay in bacteria conducted according to OECD guideline 471, Salmonella typhimurium strains TA1535, TA1537, TA98, TA100 and E. coli strain WP2uvrA were exposed to Propyl (S)-lactate (99.5% purity) at concentrations of 0, 100, 333, 1000, 3330 and 5000 µg/plate in the presence and absence of mammalian metabolic activation. The test item was tested up to the limit concentration of 5000 µg/plate. The positive controls induced the appropriate responses in the corresponding strains. There was no evidence of induced mutant colonies over background. Based on the results, the test item can be considered to be non-mutagenic.
This study is classified as acceptable and satisfies the requirement for Test Guideline OECD 471 for in vitro mutagenicity (bacterial reverse gene mutation assay).
This information is used in a read-across approach in the assessment of the target substance. For details and justification of read-across please refer to the read-across report attached to IUCLID section 13
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- For details and justification of read-across please refer to the report attached in section 13 of IUCLID.
- 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
- Remarks:
- no toxicity in experiment 1, no severe toxicity in experiment 2.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
PURASOLV® EL did not precipitate in the exposure medium up to and including the concentration of 1180 μg/ml (= 10 mM). Since testing up to 0.01 M is recommended in the guidelines, this concentration was used as the highest test substance concentration in the dose range finding test.
The pH and osmolarity of a concentration of 1180 μg/ml were 7.4 and 0.302 Osm/kg respectively (compared to 7.5 and 0.286 Osm/kg in the solvent control).
RANGE-FINDING/SCREENING STUDIES:
In the dose range finding test, L5178Y mouse lymphoma cells were treated with a test substance concentration range of 10 to 1180 µg/mL in the absence of S9-mix with a 3 and 24 hour treatment period and in the presence of S9-mix with a 3 hour treatment period.
Table 1 (Tables see attached background information) shows the cell counts of the cultures after 3 hours of treatment with various concentrations of PURASOLV® EL and after 24 and 48 hours of subculture and the calculated suspension growth and the relative suspension growth.
Both in the absence and presence of S9-mix, no toxicity in the relative suspension growth was observed up to and including the highest test substance concentration of 1180 μg/mL compared to the suspension growth of the solvent controls.
Table 2 shows the cell counts of the cultures after 24 hours of treatment with various concentrations of PURASOLV® EL and after 24 hours of subculture and the calculated suspension growth and the relative suspension growth.
In the absence of S9-mix, the relative suspension growth was 38% at the test substance concentration of 1180 µg/ml compared to the relative suspension growth of the solvent control.
COMPARISON WITH HISTORICAL CONTROL DATA:
The spontaneous mutation frequencies in the solvent-treated control cultures were between the minimum and maximum value of the historical control data range. - Conclusions:
- In conclusion, Ethyl (S)-lactate is considered to be non-mutagenic in the in vitro mammalian cell gene mutation test (OECD 476, nowadays OECD 490) in the presence and absence of mammalian metabolic activation.
- Executive summary:
In a mammalian cell gene mutation assay conducted according to OECD Guideline 476 (nowadays OECD 490), L5178Y mouse lymphoma cells cultured in vitro were exposed to Ethyl (S)-lactate (purity 99.83%) in RPMI 1640 medium at concentrations of 0, 0.3, 1, 3, 10, 33, 100, 333 and 1180 µg/mL in the presence and absence of mammalian metabolic activation. The test item was tested up to the highest concentration recommended in the guideline (0.01 M = 1180 µg/mL). Positive controls induced the appropriate response. There was no evidence of induced mutant colonies over background. Based on the results, it can be concluded, that Ethyl (S)-lactate is not mutagenic in the mouse lymphoma L5178Y test system under the experimental conditions described.
This study is classified as acceptable. This study satifies the requirement for Test Guideline OECD 490 for in vitro mutagenicity (mammalian forward gene mutation) data.
This information is used in a read-across approach in the assessment of the target substance. For justification of read-across please refer to the attached read-across report (see IUCLID section 13).
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- For details and justification of read-across please refer to the read-across report attached to IUCLID section 13.
- 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 nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: At the highest concentration of test substance (0.01 M equal to 901 µg/mL) the pH was 6.84 compared to a pH of 7.31 in the solvent control.
- Effects of osmolality: At the highest concentration of test substance (0.01 M equal to 901 µg/mL) the osmolarity was 0.319 Osm/kg compared to an osmolarity of 0.299 Osm/kg in the solvent control
- Water solubility: miscible
- Precipitation: No
RANGE-FINDING/SCREENING STUDIES:
Cytotoxicity data were obtained by treating 8 x 10^6 cells (10^6 cells/mL for 3 hours treatment) or 5 x 10^6 cells (1.25 x 10^5 cells/mL for 24 hours treatment) with 0, 17, 52, 164, 512 and 901 µg of test substance for 3 hours in the presence of S9-mix and for 3 and 24 hours in the absence of S9-mix.
After exposure, the cells were separated from treatment solutions centrifugation steps and re-suspended in RPM 1640 medium supplemented with 10% (v/v) inactivated horse serum (R10 medium). Cells were counted with the coulter particle counter.
For determination of the cytotoxicity, the surviving cells of the 3 hours treatment were subcultured twice. After 24 hours of subculturing, the cells were counted (day 1) and subcultured again for another 24 hours, after which the cells were counted (day 2). The surviving cells of the 24 hours treatment were subcultured once. After 24 hours of subculturing, the cells were counted. If less than 1.25 x 105 cells/mL were counted no subculture was performed.
The suspension growth expressed as the reduction in cell growth after approximately 24 and 48 hours or only 24 hours cell growth, compared to the cell growth of the solvent control, was used to determine an appropriate dose range for the mutagenicity tests.
COMPARISON WITH HISTORICAL CONTROL DATA:
Spontaneous mutation frequencies in the solvent-treated control cultures were between the minimum and maximum value of the historical control range.
Experiment 1 & 2: For individual results see Tables 3-5 in box 'Any other information on results incl. tables'. - Conclusions:
- In conclusion, L(+)-lactic acid is considered to be non-mutagenic in the in vitro mammalian cell gene mutation test (OECD 476, nowadays OECD 490) in the presence and absence of mammalian metabolic activation.
- Executive summary:
In a mammalian cell gene mutation assay conducted in accordance to OECD guideline 476 (nowadays OECD 490), L5178Y mouse lymphoma cells cultured in vitro were exposed to L(+)-lactic acid (90% purity), solved in RPMI 1640 medium. In the first experiment, L(+)-lactic acid was tested up to concentrations of 901 µg/mL (0.01 M, the highest concentration recommended in the guidelines) in the absence and presence of S9-mix. The incubation time was 3 hours. In the second experiment, L(+)-lactic acid was again tested up to concentrations of 901 µg/mL in the absence S9-mix. The incubation time was 24 hours. No toxicity was observed at this dose level in the absence and presence of S9 mix. The induced mutation frequency with and without metabolic activation was not increased compared to control in all tested concentrations. The positive controls did induce the appropriate response. Based on the results, it can be concluded, that L(+)-lactic acid is not mutagenic in the mouse lymphoma L5178Y test system under the experimental conditions described.
This study is classified as acceptable. This study satisfies the requirement for OECD 476 (nowadays OECD 490 for in vitro mutagenicity (mammalian forward gene mutation) data.
This information is used in a read-across approach in the assessment of the target substance. For details and justification of read-across please refer to the read-across report attached to IUCLID section 13.
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- For details and justification of read-across please refer to the read-across report attached to IUCLID section 13.
- Reason / purpose for cross-reference:
- read-across source
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No
RANGE-FINDING/SCREENING STUDIES: Dose range finding test performed with TA100 and WP2uvrA. Doses tested: 0, 3, 10, 33, 100, 333, 1000, 3330, 5000 µg/plate. Based on the results, the following doses were selected for the main experiment with TA1535, TA1537 and TA98 in the presence and absence of S9-mix: 100, 333, 1000, 330 and 5000 µg/plate.
COMPARISON WITH HISTORICAL CONTROL DATA: The negative control values were within the laboratory historical control data ranges, except for TA100 in the absence of S9-mix, second experiment. Evaluation: The mean plate count (146) was just outside the limit of the range (144) and clear negative results are observed in all experiments. Therefore, this deviation in the mean plate count of the solvent control had no effect on the results of the study. The strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly, except the response for TA1535 in the absence of S9-mix, first experiment. Evaluation: The value (257) was just below the limit of the range (262). The purpose of the positive control is as a reference for the test system, where a positive response is required to check if the test system functions correctly. Since the value was more than 3 times greater than the concurrent solvent control values, this deviation in the mean plate count of the positive control had no effect on the results of the study.
ADDITIONAL INFORMATION ON CYTOTOXICITY:
No reduction of the bacterial background lawn and no biologically relevant decrease in the number of revertants were observed.
MUTAGENICITY:
Experiment 1: No increase in the number of revertants was observed upon treatment with the test item under all conditions tested.
Experiment 2: Based on the results from the first experiment, the test item was tested up to 5000 µg/plate. No increase in the number of revertants was observed. - Conclusions:
- In conclusion, L(+)-lactic acid is not genotoxic in the bacterial reverse gene mutation assay (OECD 471) in the presence and absence of mammalian metabolic activation.
- Executive summary:
In a reverse gene mutation assay in bacteria conducted according to OECD guideline 471, Salmonella typhimurium strains TA1535, TA1537, TA98, TA100 and E. coli strain WP2uvrA were exposed to L(+)-lactic acid (90% purity) at concentrations of 0, 100, 333, 1000, 3330 and 5000 µg/plate in the presence and absence of mammalian metabolic activation.
L(+)-lactic acid was tested up to the limit concentration of 5000 µg/plate. The positive controls induced the appropriate responses in the corresponding strains. There was no evidence of induced mutant colonies over background. Based on the results, the test item can be considered to be non-mutagenic.
This study is classified as acceptable and satisfies the requirement for Test Guideline OECD 471 for in vitro mutagenicity (bacterial reverse gene mutation assay).
This information is used in a read-across approach in the assessment of the target substance. For details and justification of read-across please refer to the read-across report attached to IUCLID section 13
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- For details and justification of read-across please refer to the read-across report attached to IUCLID section 13.
- Reason / purpose for cross-reference:
- read-across source
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- 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
- Untreated negative controls validity:
- not examined
- True negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- Please see table 1 in box " Any other information on results incl. tables".
- Conclusions:
- Based on the results, the test item showed no evidence of mutagenic activity in a bacterial reverse mutation assay.
- Executive summary:
The plate-incorporation test was performed according to the standard procedure as described by Ames et al., 1975 (similar/equivalent to OECD TG 471). Methanol was assayed in the bacterial reverse mutation assay with the S. typhimurium strains TA98, TA100, TA1535, TA1537, TA1538 and E. coli strain WP2 uvrA both with and without S9 mix at a concentration of up to 5000 µg/plate. Based on the results, there was no evidence of induced mutant colonies in comparison to controls.
This information is used in a read-across approach in the assessment of the target substance. For details and justification of read-across please refer to the read-across report attached to IUCLID section 13.
Referenceopen allclose all
Table 2: Results of Salmonella mutagenicity assay with ethyl lactate
Average Revertants Per Plate**± Standard Deviation |
|||||
Liver Microsomes: None |
|||||
Dose (µg/plate) |
TA98 |
TA100 |
TA1535 |
TA1537 |
TA1538 |
0* |
14 ± 5 |
96 ± 20 |
20 ± 3 |
5 ± 1 |
13 ± 3 |
667 |
19 ± 2 |
111 ± 6 |
20 ± 6 |
5 ± 1 |
13 ± 3 |
1000 |
9 ± 3 |
104 ± 14 |
22 ± 2 |
7 ± 3 |
14 ± 4 |
3333 |
14 ± 2 |
111 ± 12 |
19 ± 1 |
5 ± 1 |
19 ± 2 |
6667 |
17 ± 3 |
107 ± 11 |
17 ± 2 |
8 ± 2 |
12 ± 2 |
10000 |
20 ± 7 |
99 ± 6 |
18 ± 5 |
5 ± 2 |
11 ± 2 |
Positive control |
519 ± 53 |
485 ± 12 |
281 ± 7 |
241 ± 23 |
599 ± 37 |
Liver Microsomes: Rat Liver S-9 |
|||||
Dose (µg/plate) |
TA98 |
TA100 |
TA1535 |
TA1537 |
TA1538 |
0* |
23 ± 3 |
105 ± 12 |
12 ± 4 |
9 ± 3 |
22 ± 2 |
667 |
34 ± 7 |
99 ± 14 |
15 ± 5 |
6 ± 2 |
20 ± 8 |
1000 |
21 ± 1 |
102 ± 9 |
13 ± 6 |
8 ± 3 |
22 ± 7 |
3333 |
27 ± 6 |
102 ± 5 |
13 ± 3 |
7 ± 3 |
21 ± 6 |
6667 |
28 ± 5 |
105 ± 13 |
13 ± 3 |
12 ± 5 |
16 ± 5 |
10000 |
28 ± 3 |
102 ± 4 |
12 ± 3 |
7 ± 2 |
21 ± 3 |
Positive control |
950 ± 36 |
2358 ± 118 |
300 ± 4 |
167 ± 7 |
1387 ± 125 |
*Vehicle control with dionized, distilled grade water
** Mean of 3 plates
All results are presented in Tables 1-8 in the attached background material (NOTOX Project 494586).
Table 1: Results of the in vitro micronucleus test
| Chemical | Dose | Number of MN/1000 cells (mean +/- SE) |
|||
| Medium | 50 µL/mL | 4.00 +/- 0.71 | |||
| Methanol | 50 µl/mL | 3.50 +/- 1.19 | |||
| MMS | 100 µg/mL | 16.5 +/- 0.50 | |||
| EMS | 100 µg/mL | 7.0 +/- 0.69 | |||
| MMNG | 0.08 µg/mL | 8.2 +/- 0.83 | |||
Mitotic Indices:
Table1: Mitotic index of human lymphocyte cultures treated with L(+)-lactic acid in the dose range finding test
L(+)-lactic acid concentration (µg/ml) |
Number of metaphases:
Absolute |
Number of metaphases:
Number of cells scored |
Number of metaphases:
Percentage of control |
Without metabolic activation (-S9-mix) |
|
|
|
3 h exposure time, 24 h fixation time |
|
|
|
Control a) |
96 |
1004 |
100 |
10 |
99 |
1007 |
103 |
33 |
80 |
1045 |
83 |
100 |
60 |
1008 |
63 |
333 |
66 |
1009 |
69 |
901 |
63 |
1003 |
66 |
24 h exposure time, 24 h fixation time |
|
|
|
Control a) |
65 |
1007 |
100 |
10 |
62 |
1042 |
95 |
33 |
71 |
1041 |
109 |
100 |
66 |
1016 |
102 |
333 |
68 |
1048 |
105 |
901 |
36 |
1028 |
55 |
48h exposure time,48h fixation time |
|
|
|
Control a) |
68 |
1017 |
100 |
10 |
64 |
1026 |
94 |
33 |
51 |
1013 |
75 |
100 |
65 |
1010 |
96 |
333 |
57 |
1017 |
84 |
901 |
33 |
1033 |
49 |
With metabolic activation (+S9-mix) |
|
|
|
3 h exposure time, 24 h fixation time |
|
|
|
Control a) |
85 |
1044 |
100 |
10 |
70 |
1013 |
82 |
33 |
71 |
1008 |
84 |
100 |
68 |
1006 |
80 |
333 |
66 |
1020 |
78 |
901 |
71 |
1007 |
84 |
a.) culture medium
Table 2: Mitotic index of human lymphocyte cultures treated with L(+)-lactic acid in the first cytogenetic assay
L(+)-lactic acid concentration (µg/ml) |
Number of metaphases a)
Absolute |
Number of metaphases a)
Number of cells scored |
Percentage of control
|
Without metabolic activation (-S9-mix) |
|
|
|
3 h exposure time, 24 h fixation time |
|
|
|
Control b) |
3-36 |
1009-1028 |
100 |
10 |
3-35 |
1002-1012 |
103 |
100 |
3-28 |
1008-1033 |
90 |
901 |
1-9 |
1002-1040 |
34 |
MMC-C; 0.5 µg/ml |
4-7 |
1026-1031 |
16 |
MMC-C; 0.75 µg/ml |
7-5 |
1029-1004 |
17 |
With metabolic activation (+S9-mix) |
|
|
|
3 h exposure time, 24 h fixation time |
|
|
|
Control b) |
3-48 |
1040-1028 |
100 |
10 |
3-27 |
1013-1007 |
79 |
100 |
4-35 |
1035-1001 |
101 |
901 |
4-32 |
1008-1007 |
93 |
CP; 10 µg/ml |
21-14 |
1005-1025 |
43 |
a) Duplicate cultures
b) Culture medium
Table 3: Mitotic index of human lymphocyte cultures treated with L(+)-lactic acid in the second cytogenetic assay
L(+)-lactic acid concentration (µg/ml) |
Number of metaphasesa
Absolute |
Number of metaphasesa
Number of cells scored |
Percentage of control |
Without metabolic activation (-S9-mix) |
|
|
|
24 h exposure time, 24 h fixation time |
|
|
|
Control b) |
90-85 |
1000-1000 |
100 |
100 |
75-83 |
1000-1003 |
90 |
333 |
67-65 |
1008-1000 |
75 |
666 |
73-66 |
1001-1000 |
79 |
901 |
39-42 |
1002-1000 |
46 |
MMC-C; 0.2 µg/ml |
24-34 |
1000-1003 |
33 |
MMC-C; 0.3 µg/ml |
21-33 |
1003-1000 |
31 |
48 h exposure time, 48 h fixation time |
|
|
|
Control b) |
93-88 |
1005-1000 |
100 |
100 |
71-87 |
1001-1000 |
87 |
333 |
66-51 |
1000-1000 |
65 |
666 |
34-37 |
1000-1002 |
39 |
901 |
22-24 |
1003-1000 |
25 |
MMC-C; 0.1 µg/ml |
18-20 |
1002-1003 |
21 |
MMC-C; 0.15 µg/ml |
17-19 |
1000-1004 |
20 |
With metabolic activation (+S9-mix) |
|
|
|
3 h exposure time, 48 h fixation time |
|
|
|
Control b) |
88-87 |
1000-1000 |
100 |
10 |
66-75 |
1000-1045 |
81 |
100 |
62-64 |
1003-1005 |
72 |
901 |
71-63 |
1000-1000 |
77 |
CP; 10 µg/ml |
22-18 |
1005-1000 |
-c.) |
a) Duplicate cultures
b) Culture medium
c) CP was fixed after 24 hours. Therefore, the mitotic index could not be calculated as percentage of control.
Chromosome aberrations in human lymphocyte cultures treated with L(+)-lactic acid
1. Cytogenetic Assay:
| Conc | Exposure Time [h] | Fixation Time [h] | Metabolic Activation | Culture | Mitotic Index [%] | Cells scored |
Cells + gaps |
Cells - gaps |
| Culture medium | 3 | 24 | without | A+B | 100 | 200 | 0 | 0 |
| 10 µg/ml | 3 | 24 | without | A+B | 103 | 200 | 5 | 5 |
| 100 µg/ml | 3 | 24 | without | A+B | 90 | 200 | 0 | 0 |
| 901 µg/ml | 3 | 24 | without | A+B | 34 | 200 | 4 | 4 |
| MMC-C 0.5 µg/ml | 3 | 24 | without | A+B | 16 | 150 | 76*** | 76*** |
| Culture medium | 3 | 24 | with | A+B | 100 | 200 | 2 | 2 |
| 10 µg/ml | 3 | 24 | with | A+B | 79 | 200 | 3 | 3 |
| 100 µg/ml | 3 | 24 | with | A+B | 101 | 200 | 2 | 2 |
| 901 µg/ml | 3 | 24 | with | A+B | 93 | 200 | 2 | 2 |
| CP 10 µg/ml | 3 | 24 | with | A+B | 43 | 200 | 59*** | 59*** |
*) Significantly different from control group (Chi-square test), p < 0.001
2. Cytogenetic Assay:
| Conc | ExposureTime [h] | FixationTime [h] | Metaboic Activation | Culture | Mitotic Index [%] |
Cells scored |
Cells + gaps |
Cells - gaps |
| Culture medium | 3 | 48 | with | A+B | 100 | 200 | 2 | 1 |
| 10 µg/ml | 3 | 48 | with | A+B | 81 | 200 | 1 | 1 |
| 100 µg/ml | 3 | 48 | with | A+B | 72 | 200 | 0 | 0 |
| 10 µg/ml | 3 | 48 | with | A+B | 77 | 200 | 3 | 3 |
| CP 10 µg/ml | 3 | 48 | with | A+B | n.d.b | 100 | 53*** | 53*** |
| Culture medium | 24 | 24 | without | A+B | 100 | 200 | 1 | 1 |
| 100 µg/ml | 24 | 24 | without | A+B | 90 | 200 | 1 | 1 |
| 666 µg/ml | 24 | 24 | without | A+B | 79 | 200 | 0 | 0 |
| 901 µg/ml | 24 | 24 | without | A+B | 46 | 200 | 3 | 3 |
| MMC- 0.1 µg/ml | 24 | 24 | without | A+B | 33 | 100 | 53*** | 53*** |
| Culture medium | 48 | 48 | without | A+B | 100 | 200 | 1 | 1 |
| 100 µg/ml | 48 | 48 | without | A+B | 87 | 200 | 3 | 3 |
| 333 µg/ml | 48 | 48 | without | A+B | 65 | 200 | 2 | 2 |
| 666 µg/ml | 48 | 48 | without | A+B | 39 | 200 | 2 | 2 |
| MMC- 0.1 µg/ml | 48 | 48 | without | A+B | 21 | 100 | 53*** | 53*** |
bCP was fixed after 24 hours. Therefore, the mitotic index could not be calculated as percentage of control.
*) Significantly different from control group (Chi-square test), p < 0.001
Precipitate:
The test substance did not precipitate in the top agar. Precipitation of the test substance on the plates was not observed at the start or at the end of the incubation period in all tester strains.
Toxicity:
The bacterial background lawn was not reduced at any of the concentrations tested and no biologically relevant decrease in the number of revertants was observed.
Number of revertants:
All bacterial strains showed negative responses over the entire dose range. The negative and positive controls were within the historical control data ranges indicating that the test conditions were adequate.
Tables are given in the attached background information.
Table 1: Dose-range finding test: Cytotoxicity of L(+)-lactic acid (3 hours treatment)
| Dose (µg/mL) |
Cell count after 3 hours of treatment (cells/mL x 10^5) | Cell count after 24 hours of subculture (cells/mL x 10^5) | Cell count after 48 hours of subculture (cells/mL x 10^5) | SG(1) x 10^5 cells/mL) |
RSG (2) % |
without metabolic activation |
|||||
| SC | 6.9 | 5.0 | 6.9 | 152 | 100 |
| 17 | 6.3 | 5.0 | 7.4 | 149 | 98 |
| 52 | 7.0 | 5.2 | 6.9 | 161 | 106 |
| 164 | 7.3 | 5.2 | 7.1 | 173 | 113 |
| 512 | 7.6 | 5.2 | 6.8 | 172 | 113 |
| 901 | 6.8 | 5.4 | 7.0 | 166 | 109 |
with metabolic activation |
|||||
| SC | 5.3 | 4.9 | 7.8 | 130 | 100 |
| 17 | 5.2 | 5.2 | 7.5 | 130 | 100 |
| 52 | 4.2 | 5.2 | 7.6 | 106 | 82 |
| 164 | 4.1 | 5.3 | 7.2 | 100 | 77 |
| 512 | 5.0 | 5.1 | 7.5 | 122 | 94 |
| 901 | 4.3 | 5.1 | 7.4 | 104 | 80 |
Note: all calculations were made without rounding off
SC = solvent control = exposure medium
(1) = suspension growth
(2) relative suspension growth
SG= (Cell count after 3 h treatment) x (Cell count after 24 h subculture)/(Cells subcultured (at t=3 h)(1.25x10^5 c/mL)) x (Cell count after 48 h subculture)/(Cells subcultured (at t=24 h) (1.25 x 10^5 c/mL))
RSG = [SG(test)/SG(control)] x 100
Table 2: Dose-range finding test: Cytotoxicity of L(+)-lactic acid (24 hours treatment)
| Dose (µg/mL) |
Cell count after 24 hours of treatment (cells/mL x 10^5) | Cell count after 24 hours of subculture (cells/mL x 10^5) | SG(1) x 10^5 cells/mL) |
RSG (2) % |
without metabolic activation |
||||
SC |
9.5 | 5.9 | 45 | 100 |
| 17 | 8.9 | 5.9 | 42 | 93 |
| 52 | 9.3 | 5.7 | 42 | 93 |
| 164 | 9.1 | 5.2 | 39 | 85 |
| 512 | 8.8 | 5.5 | 39 | 87 |
| 901 | 7.2 | 4.6 | 26 | 58 |
Note: all calculations were made without rounding off
SC = solvent control = exposure medium
(1) = suspension growth
(2) relative suspension growth
SG = (Cell count after 24 h treatment) x (Cell count after 24 h subculture)/(Cells subcultured after treatment (1.25 x 10^5 c/mL)
RSG = [SG(test)/SG(control)] x 100
Cytotoxic and mutagenic response of L(+)-lactic acid in the mouse lymphoma L5178Y test system
Abbreviations:
RSG: Relative Suspension Growth
CE: Cloning Efficiency
RS: Relative Survival
RTG: Relative Total Growth
MF: Mutation Frequency per 10^6 Survivors
SC: Solvent Control (= Exposure Medium)
MMS: Methylmethanesulfonate
CP: Cyclophosphamide
Experiment 1
Table 3: 3 h treatment, without metabolic activation
| Dose [µg/mL] |
RSG [%] |
CEday2 [%] |
RSday2 [%] |
RTG [%] |
MF total |
MF small |
MS large |
| SC1 | 100 | 97 | 100 | 100 | 89 | 70 | 16 |
| SC2 | 100 | 80 | 100 | 100 | 86 | 66 | 18 |
| 0.54 | 107 | 86 | 98 | 105 | 98 | 71 | 25 |
| 1.7 | 118 | 79 | 89 | 106 | 98 | 72 | 23 |
| 5.4 | 126 | 83 | 93 | 117 | 94 | 62 | 28 |
| 17 | 129 | 77 | 87 | 112 | 122 | 91 | 26 |
| 52 | 108 | 75 | 84 | 91 | 124 | 97 | 23 |
| 164 | 112 | 78 | 88 | 99 | 104 | 66 | 34 |
| 512 | 106 | 72 | 82 | 87 | 147 | 99 | 41 |
| 901 | 101 | 88 | 99 | 100 | 116 | 89 | 23 |
| MMS | 79 | 41 | 47 | 37 | 1149 | 870 | 191 |
Table 4: 3 h treatment, with metabolic activation
| Dose [µg/mL] |
RSG [%] |
CEday2 [%] |
RSday2 [%] |
RTG [%] |
MF total |
MF small |
MS large |
| SC1 | 100 | 68 | 100 | 100 | 51 | 26 | 23 |
| SC2 | 100 | 64 | 100 | 100 | 55 | 30 | 25 |
| 0.54 | 92 | 78 | 118 | 108 | 53 | 31 | 20 |
| 1.7 | 78 | 111 | 168 | 132 | 26 | 17 | 8 |
| 5.4 | 56 | 93 | 140 | 79 | 38 | 28 | 10 |
| 17 | 60 | 97 | 146 | 88 | 31 | 22 | 9 |
| 52 | 65 | 66 | 100 | 65 | 62 | 47 | 14 |
| 164 | 92 | 74 | 111 | 102 | 53 | 45 | 7 |
| 512 | 64 | 77 | 116 | 74 | 45 | 12 | 32 |
| 901 | 93 | 81 | 123 | 114 | 45 | 25 | 19 |
| CP | 37 | 29 | 43 | 16 | 849 | 647 | 167 |
Experiment 2
Table 5: 24 h treatment, without metabolic activation
| Dose [µg/mL] |
RSG [%] |
CEday2 [%] |
RSday2 [%] |
RTG [%] |
MF total |
MF small |
MS large |
| SC1 | 100 | 98 | 100 | 100 | 57 | 26 | 30 |
| SC2 | 100 | 102 | 100 | 100 | 50 | 19 | 30 |
| 0.54 | 92 | 84 | 84 | 77 | 63 | 19 | 42 |
| 1.7 | 91 | 86 | 86 | 78 | 71 | 40 | 28 |
| 5.4 | 99 | 89 | 89 | 88 | 65 | 25 | 38 |
| 17 | 90 | 89 | 89 | 80 | 49 | 11 | 38 |
| 52 | 86 | 98 | 98 | 84 | 51 | 16 | 34 |
| 164 | 85 | 88 | 87 | 74 | 72 | 34 | 36 |
| 512 | 78 | 90 | 90 | 71 | 50 | 22 | 26 |
| 901 | 64 | 107 | 107 | 68 | 53 | 9 | 43 |
| MMS | 80 | 61 | 61 | 49 | 621 | 198 | 368 |
Table 1: Results of the Ames test
| Compound | Dose (µg/plate) | Revertrant colonies/plate | |||||||||||||
| TA100 | TA1535 | E. coli WP2 uvrA | TA98 | TA1537 | TA1538 | ||||||||||
| -S9 | +S9 | -S9 | +S9 | -S9 | +S9 | -S9 | +S9 | -S9 | +S9 | -S9 | +S9 | ||||
| H2O (neg. control) | - | 149 +/- 17.1 | 161 +/- 16.2 | 28 +/- 6.9 | 15 +/- 3.6 | 32 +/- 7.3 | 33 +/- 10.3 | 29 +/- 6.2 | 39 +/- 8.6 | 16 +/- 6.4 | 21 +/- 8.1 | 21 +/- 5.5 | 28 +/- 7.0 | ||
| AF-2 | 0.01 | 501 +/- 84.7 | |||||||||||||
| 0.05 | 1082 +/- 293.7 | 278 +/- 64.8 | |||||||||||||
| EENG | 5.0 | 1101 +/- 683.1 | |||||||||||||
| 9AC | 80.0 | 889 +/- 275.7 | |||||||||||||
| 4NQO | 0.25 | 270 +/- 66.2 | |||||||||||||
| B [a]P | 5.0 | 1084 +/- 236.3 | 809 +/- 108.4 | 313 +/- 41.6 | 354 +/- 89.4 | ||||||||||
| 2AA | 5.0 | 440 +/- 198.6 | 359 +/- 127.0 | ||||||||||||
| Methanol | 5.0 | 146 | 162 | 22 | 16 | 31 | 29 | 28 | 38 | 18 | 35 | 14 | 31 | ||
| 10.0 | 155 | 164 | 19 | 7 | 36 | 43 | 31 | 37 | 22 | 29 | 17 | 27 | |||
| 50.0 | 155 | 180 | 35 | 11 | 28 | 27 | 42 | 33 | 19 | 29 | 13 | 24 | |||
| 100.0 | 175 | 161 | 24 | 13 | 30 | 36 | 36 | 37 | 22 | 32 | 16 | 14 | |||
| 500.0 | 149 | 177 | 21 | 10 | 32 | 32 | 36 | 32 | 21 | 26 | 18 | 18 | |||
| 1000.0 | 159 | 155 | 19 | 16 | 25 | 33 | 27 | 39 | 19 | 29 | 17 | 16 | |||
| 5000.0 | 167 | 154 | 20 | 17 | 36 | 31 | 34 | 37 | 22 | 30 | 17 | 20 | |||
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Due to the enzymatically catalyzed hydrolysis of Methyl (S)-lactate into methanol and L-lactic acid, the toxicology of Methyl (S)-lactate can be understood in terms of the toxicology of the metabolites. Lactic acid is a ubiquitous and integral part of mammalian metabolism and therefore of minor toxicological relevance in comparison to methanol which is, as an alcohol, more important for the toxicological assessment. No data is available for the target substance itself. Thus, data from the suitable read-across partners lactic acid, methanol and from the structurally related ethyl lactate, ethyl (S)-lactate and propyl (S)-lactate were used in a weight-of-evidence approach to assess the genotoxic potential of the target substance. All read-across partners were tested as not genotoxic.
Based on the available data from suitable read-across partners, the target substance can be considered to be non-genotoxic.
Justification for classification or non-classification
Based on the available results, the target substance Methyl (S)-lactate is not considered to be genotoxic and no classification is warranted in accordance with CLP Regulation 1272/2008.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
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