Disodium adipate

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian germ cell study: cytogenicity / chromosome aberration

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1974

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No GLP but overall good documentation, purity not specified, no positive control for every experiment.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1974

Materials and methods

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 C-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

Type of assay:

chromosome aberration assay

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

Details on test material:

IUCLID4 Test substance: purity not specified

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

Remarks:

Doses / Concentrations:
Test 1: acute and subacute: 3.75, 37.5, 375 mg/kg bw/day; Test 2: acute 5000 mg/kg bw and subacute 2500 mg/kg bw/day
Basis:

No. of animals per sex per dose:

Groups of  5 treated and 3 control animals were used.

Examinations

Tissues and cell types examined:

bone marrow

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 I (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 (triethylene melamine 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.

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.

In summary, adipic acid can be considered non-mutagenic as
measured by the cytogenetic test.

Applicant's summary and conclusion

Executive summary:

Adipic acid was not mutagenic in in vivo cytogenetic studies where groups of five male rats were gavaged with adipic acid doses up to 5000 mg/kg bw (acute studies) and with doses up to 2500 mg/kg bw/day (five-days subacute studies). 200 to 500 metaphase chromosomes of bone marrow cells per dose were scored for chromatid gaps and breaks, chromosome gaps and breaks, reunions, cells with greater than ten aberrations, polyploidy, pulverization and other chromosomal aberrations. The mitotic indices for all dose groups were considered to be within the normal limits of the controls and there was no evidence of chromosomal damage. The positive control groups, performed only during the acute studies, were functional (Litton Bionetics, Inc. 1974).