Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study without detailed documentation

Data source

Reference
Reference Type:
publication
Title:
Genotoxicity and glucose tolerance induction by acetyltriethylcitrate, substitute plasticizer compared to di(2-ethylhexyl)phthalate
Author:
Jae-Wook Lee, Seok Jong Lee, Myung Chan Gye, Eun-Yi Moon
Year:
2019
Bibliographic source:
Sci Rep 9, 12237

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
GLP compliance:
not specified
Remarks:
Data from Korea and not performed for REACH registration, GLP is not specified in the publication
Type of assay:
mammalian erythrocyte micronucleus test

Test material

Constituent 1
Chemical structure
Reference substance name:
Triethyl O-acetylcitrate
EC Number:
201-066-5
EC Name:
Triethyl O-acetylcitrate
Cas Number:
77-89-4
Molecular formula:
C14H22O8
IUPAC Name:
triethyl 2-acetoxypropane-1,2,3-tricarboxylate
Details on test material:
Santa Cruz Biotechnology Inc, Dallas, TX, USA

Test animals

Species:
mouse
Strain:
CD-1
Details on species / strain selection:
Specific pathogen-free (SPF) seven weeks old male CD-1 mice obtained from ORIENTBIO INC. (Sungnam, Republic of Korea). All animals were acclimated for 7 days and observed daily for general health.
Sex:
male
Details on test animals or test system and environmental conditions:
Five mice were housed in the transparent acrylic cage and maintained in the pathogen-free authorized facility in WOOJUNG BIO CROWISE or in Sejong University where the temperature was at 20–22 °C, the humidity at 50–60%, and a dark/ light cycle at 12 h. Mouse-used all experiments were carried out in strict accordance with the guidelines by the recommendations in the Guide for the Care and Use of Laboratory Animals of ‘Animal and Plant Quarantine Agency’, Republic of Korea.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
carboxymethylcellulose (CMC: 0.5%
Duration of treatment / exposure:
24 h
Frequency of treatment:
1
Post exposure period:
2 h, day 1, 2 and 3
Doses / concentrationsopen allclose all
Dose / conc.:
500 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Dose / conc.:
2 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
Positive control(s):
Positive control group: intraperitonelly injection with 2 mg/kg MMC dissolved in saline solution.

Examinations

Tissues and cell types examined:
bone marrow cells from femur canal
Evaluation criteria:
Index of bone marrow cytotoxicity was calculated by the ratio of PCE to the total number of erythrocytes.
Statistics:
Criteria of Kastenbaum and Bowman was used to analyze the significant positive increase in the number of MNPCE33. Homogeneity of variance in the frequency of PCE and body weight was analyzed by using
Bartlett’s test. In addition, one-way analysis of variance (ANOVA) was employed for homogeneous data; then, if significant, Dunnett’s t-test was applied for multiple comparisons. P value of <0.05 or <0.01 was considered to
be significant.

Results and discussion

Test results
Sex:
male
Genotoxicity:
negative
Toxicity:
no effects
Vehicle controls validity:
valid
Negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
While no significant incidence of MNPCE in PCE was observed in groups treated with ATEC, it was significantly increased in MMC-treated group compared to control group. No statistically significant differences in the ratio of PCE among total erythrocytes were noted at 24, 48 and 72 h after administration of ATEC or MMC compared to control group.

Applicant's summary and conclusion

Conclusions:
ATEC did not induce micronuclei formation in PCE from mouse bone marrow under the experimental conditions.
Executive summary:

Micronuclei formation was tested in male CD-1 mice (OECD 474). Mice were treated with 500, 1000 and 2000 mg/kg by oral gavage. Incidence of MNPCE in PCE and ratio of PCE among total erythrocytes was determined as compared to those in control group. MMC was used as a positive control.