Potassium (E,E)-hexa-2,4-dienoate

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study (GLP)

Justification for type of information:

Since the sorbate anion in the (eco-) toxicologically relevant moiety and its presence in dissociated or non-dissociated form is entirely determined by pH, extrapolation to potassium sorbate, the substance in question, is not considered to be restricted.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1989

Report date:

1989

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Version / remarks:

1983

Deviations:

no

GLP compliance:

yes

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

Hexa-2,4-dienoic acid

EC Number:

203-768-7

EC Name:

Hexa-2,4-dienoic acid

Cas Number:

110-44-1

Molecular formula:

C6H8O2

IUPAC Name:

hexa-2,4-dienoic acid

Details on test material:

- Name of test material: Sorbic acid
- Physical state: solid, white powder
- Analytical purity: 99 %
- Stability under test conditions: The test substance was considered to be stable for the duration of the study.
- Storage condition of test material: dark at 4 °C

Test animals

Species:

mouse

Strain:

NMRI

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Hoechst AG, Frankfurt, Germany
- Strain: NMRKf (SPF71)
- Age at study initiation: 7 weeks
- Weight at study initiation: males: 27–36 g, females: 21–29 g
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2 °C
- Humidity (%): 55 ± 10 %
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: sesame oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency): freshly prepared every day

Duration of treatment / exposure:

72 hours

Frequency of treatment:

single application

Post exposure period:

24, 48 or 72 hours

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day

Remarks:

Basis: actual ingested

Doses / concentrations 2

Dose / conc.:

500 mg/kg diet

Remarks:

Basis: actual ingested

Doses / concentrations 3

Dose / conc.:

1 500 mg/kg bw/day

Remarks:

Basis: actual ingested

Doses / concentrations 4

Dose / conc.:

5 000 mg/kg diet

Remarks:

Basis: actual ingested
Criteria for selection of MTD: a preliminary study reported 5000 mg/kg bw as the highest tolerable dose

No. of animals per sex per dose:

5 males and 5 females

Control animals:

yes, concurrent vehicle

Positive control(s):

Positive control: 50 mg/kg bw Cyclophosphamide (Endoxan ®), concentration: 50 mg/kg bw.

Examinations

Tissues and cell types examined:

Bone marrow:
- Number of animals: 5 per sex at each of 3 time points
- Number of cells: 1000
- Time points: 24, 48 and 72 hours after treatment
- Type of cells: Erythrocytes in bone marrow
- Parameters: Polychromatic/normo-chromatic erythrocyte ratio, occurrence of micronuclei

Evaluation criteria:

Criteria for evaluating results: number of cells with micronuclei, PCE:NCE ratio

Statistics:

Statistical methods: Wilcoxon

Results and discussion

Test results

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

not specified

Vehicle controls validity:

valid

Remarks:

Sesame oil

Negative controls validity:

not specified

Positive controls validity:

valid

Remarks:

Cyclophosphamide (Endoxan ®)

Additional information on results:

RESULTS OF DEFINITIVE STUDY
- No mortalities were observed, and the only clinical sign was uncoordinated gait, which had recided 24 hours after application. 
- There were no substance-related findings upon necropsy, or effects on body weight.
- Ratio of PCE/NCE (for Micronucleus assay): The PCE/NCE ratio was unaffected.
- Statistical evaluation: The test yielded no statistically significant increase of micronucleated polychromatic erythrocytes and normochromatic erythrocytes with micronuclei.

Any other information on results incl. tables

Table1: In-vivomicronucleus test of bone marrow cells from male mice receiving Sorbic acid, sesame oil (vehicle control) or Cyclophosphamide (positive control).

	Positive control	Vehicle control			Low dose (500 mg/kg)			Mid dose (1500 mg/kg)			High dose (5000 mg/kg)
Number of cells evaluated	5000	5000			5000			5000			5000
Sampling time [h]	24	24	48	72	24	48	72	24	48	72	24	48	72
No. of erythrocytes
Normo-chromatic with micronuclei	2	1	1	1	2	1	2	1	1	1	1	1	1
Polychromatic with micronuclei	32*	2	1	0	2	2	2	2	2	1	1	1	2
Ratio of erythrocytes
Polychromatic/ normo-chromatic	0.97	0.93	0.90	0.94	0.98	0.89	1.02	0.98	0.90	1.12	1.05	0.95	0.99
*) significantly different from control (p < 0.05)

 

 

Table2: In-vivomicronucleus test of bone marrow cells from female mice receiving Sorbic acid, sesame oil (vehicle control) or Cyclophosphamide (positive control).

	Positive control	Vehicle control			Low dose (500 mg/kg)			Mid dose (1500 mg/kg)			High dose (5000 mg/kg)
Number of cells evaluated	5000	5000			5000			5000			5000
Sampling time [h]	24	24	48	72	24	48	72	24	48	72	24	48	72
No. of erythrocytes
Normo-chromatic with micronuclei	1	1	0	1	0	1	1	0	1	0	1	0	0
Polychromatic with micronuclei	21*	1	2	1	1	1	3	1	1	1	2	2	1
Ratio of erythrocytes
Polychromatic/ normo-chromatic	0.89	0.99	1.17	1.00	0.94	1.12	1.14	0.94	1.22	1.24	1.13	1.05	1.15
*) significantly different from control (p < 0.05)

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative
Treatment with Sorbic acid did not cause any increase in the number of micro-nucleated polychromatic erythrocytes and micro-nucleated normo-chromatic erythrocytes at any of the three sampling times at either dose level. Statistically significant increases in the number of polychromatic cells with micronuclei were observed in positive control animals treated with 50 mg/kg bw Cyclophosphamide. Sorbic acid was not genotoxic under the conditions of this test.