Sebacic acid

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

pre-GLP, purity not specified, no positive control for every experiment

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1974

Report date:

1974

Reference 2

Reference Type:

publication

Title:

Mutagenic Evaluation of Compound FDA 71- 50, Adipic Acid

Author:

Litton Bionetics, Inc.

Year:

2001

Bibliographic source:

cited in: OECD SIDS, dicarboxylic acid category, 2001

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

Groups of 5 treated and 3 control animals were used. Animals were killed 6, 24 and 48 hours after a single administration in the acute study. In the subacute study 5 doses, 24 hours apart, were administered and animals were killed 6 hours after the last dose. Four hours after the last compound administration, and two hours prior to killing, each animal was given 4 mg/kg bw of colcemid intraperitoneally in order to arrest the bone marrow cells in mitosis. The marrow "plug" was removed and aspirated into Hanks' balanced salt solution. The specimen were centrifuged and resuspended in hypotonic 0.5% KCl. The specimens were placed in a 37 degree Celsius water bath in order to swell the cells. Following centrifugation the cells were resuspended in a fixative (3:1 absolute methanol : glacial acetic acid) and again centrifuged. Cells were resuspended and placed at 4 degree Celsius overnight. The following day cells were again centrifuged and freshly prepared fixative was added. The suspension was dropped onto a slide and ignited by an alcohol burner and allowed to flame. Slides were stained with 5% Giemsa solution. The preparations were examined by microscopy. The chromosomes of each cell were counted and only diploid cells were analyzed. They were scored for chromatid gaps and breaks, chromosome gaps and breaks, reunions, cells with greater than ten aberrations, polyploidy, pulverization, and other chromosomal aberrations which were observed. Fifty metaphase spreads were scored per animal. Mitotic indices were obtained by counting at least 500 cells and the ratio of the number of cells in mitosis / the number of cells observed was expressed as the mitotic index. Negative and positive (TEM) controls were run in each experiment. Two tests were performed at different time intervals.

GLP compliance:

no

Remarks:

pre-GLP

Type of assay:

mammalian bone marrow chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

Adipic acid

EC Number:

204-673-3

EC Name:

Adipic acid

Cas Number:

124-04-9

Molecular formula:

C6H10O4

IUPAC Name:

adipic acid

Test animals

Species:

rat

Strain:

not specified

Sex:

male

Administration / exposure

Route of administration:

oral: gavage

Duration of treatment / exposure:

Acute study: single dosing
Subacute study: once a day for 5 consecutive days

Frequency of treatment:

Acute study: single dosing
Subacute study: once a day for 5 consecutive days

Post exposure period:

Animals were killed 6, 24 and 48 hours after a single administration in the acute study. 
In the subacute study 5 doses, 24 hours apart, were administered and animals were killed 6 hours after the last dose. 

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

3.75 mg/kg bw/day

Remarks:

acute and subacute study

Doses / concentrations 2

Dose / conc.:

37.5 mg/kg bw/day

Remarks:

acute and subacute study

Doses / concentrations 3

Dose / conc.:

375 mg/kg bw/day

Remarks:

acute and subacute study

Doses / concentrations 4

Dose / conc.:

2 500 mg/kg bw/day

Remarks:

subacute study

Doses / concentrations 5

Dose / conc.:

5 000 mg/kg bw/day

Remarks:

acute study

No. of animals per sex per dose:

Groups of 5 treated/dose and 3 control male animals were used.

Control animals:

yes, concurrent vehicle

Positive control(s):

Triethylenemelamine (TEM)

Examinations

Tissues and cell types examined:

Bone marrow

Details of tissue and slide preparation:

DETAILS OF SLIDE PREPARATION:
Four hours after the last compound administration, and two hours prior to sacrifice, each animal was given 4 mg/kg bw of colcemid intraperitoneally in order to arrest the bone marrow cells in mitosis. The marrow "plug" was removed and aspirated into Hanks' balanced salt solution. The specimen were centrifuged and resuspended in hypotonic 0.5 % KCl. The specimens were placed in a 37 °C water bath in order to swell the cells. Following centrifugation the cells were resuspended in a fixative (3:1 absolute methanol : glacial acetic acid) and again centrifuged. Cells were resuspended and placed at 4 °C overnight. The following day cells were again centrifuged and freshly prepared fixative was added. The suspension was dropped onto a slide and ignited by an alcohol burner and allowed to flame. Slides were stained with 5 % Giemsa solution.

METHOD OF ANALYSIS:
The preparations were examined by microscopy. The chromosomes of each cell were counted and only diploid cells were analyzed. They were scored for chromatid gaps and breaks, chromosome gaps and breaks, reunions, cells with greater than ten aberrations, polyploidy, pulverization, and other chromosomal aberrations which were observed. Fifty metaphase spreads were scored per animal. Mitotic indices were obtained by counting at least 500 cells and the ratio of the number of cells in mitosis / the number of cells observed was expressed as the mitotic index.

Results and discussion

Test results

Sex:

male

Genotoxicity:

negative

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Test 1 (3.75, 37.5 and 375 mg/kg bw/day dosing):

Acute study: The negative control group cells contained no aberrations. The compound produced no aberrations except for one cell containing a break in the 6-hour sample of the intermediate dose level. The expected severe chromosomal damage was observed for the positive control group (triethylenemelamine treated animals). The mitotic indices were within normal limits. Negative and positive controls were functional.

Subacute study (5 days): The negative control group and the low level test group contained no aberration. The intermediate level contained one cell with a reunion and one cell that was polyploid. The highest level contained three cells with breaks and one fragment. These were considered to be within the normal limits of the historical negative controls of the laboratory. Negative control was functional. No positive control was performed.

Test 2:

Acute study: Adipic acid was administered at a single dose of 5000 mg/kg bw. The compound produced no aberrations except for 3 cells with polyploidy (2 in the 6-hour sample and 1 in the 24-hour). Neither the variety nor the number of these aberrations differed significantly from the negative controls (polyploidy observed in 4 cells). Negative and positive controls were functional.

Subacute study (5 days, 2500 mg/kg bw/day). Only 218 metaphases have been evaluated. The compound produced no aberrations except for 1 cell with polyploidy. Polyploidy was also observed in the negative control group. These are considered to be within the normal limits of the historical negative controls. Negative control was functional, no positive control.

Applicant's summary and conclusion

Conclusions:

In summary, the test substance can be considered non-mutagenic in the cytogenetic test.