Registration Dossier

Administrative data

Endpoint:
in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
other: publication
Adequacy of study:
key study
Study period:
1989
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Sufficient information is available for the interpretation of results.
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to other study

Data source

Reference
Reference Type:
publication
Title:
Acute cytogenetic effects of tyramine, MTCAs, NaCl and soy sauce on rat bone marrow cells in vivo
Author:
Fujie et al.
Year:
1989
Bibliographic source:
published in Mutation Research, 240 (1990) 281-288

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 475 (Mammalian Bone Marrow Chromosome Aberration Test)
Deviations:
not specified
Principles of method if other than guideline:
none
GLP compliance:
not specified
Type of assay:
chromosome aberration assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Sodium chloride
EC Number:
231-598-3
EC Name:
Sodium chloride
Cas Number:
7647-14-5
Molecular formula:
ClNa
IUPAC Name:
sodium chloride
Constituent 2
Reference substance name:
231-589-3
IUPAC Name:
231-589-3
Details on test material:
- Name of test material (as cited in study report): Sodium Chloride
- Molecular formula (if other than submission substance): NaCl

Test animals

Species:
rat
Strain:
Long-Evans
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source:Long-Evans rats outbred from the colony of the Ben May Laboratory for Cancer Research, University of Chicago)
- Age at study initiation: aged 4-6 weeks
- Weight at study initiation: 50-100 g
- Assigned to test groups randomly: [no/yes, under following basis: ]
- Fasting period before study: not specified in the report
- Housing:not specified in the report
- Diet (e.g. ad libitum): They were fed a commercial ration of NMF (Oriental Ferment Co., Tokyo)
- Water (e.g. ad libitum): water ad libitum
- Acclimation period: not specified in the report

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ° C.
- Humidity (%):not specified in the report
- Air changes (per hr):not specified in the report
- Photoperiod (hrs dark / hrs light):not specified in the report

Administration / exposure

Route of administration:
intraperitoneal
Vehicle:
- Vehicle(s)/solvent(s) used: physiological saline.
- Concentration of test material in vehicle: 10-40 mmole/kg (584.4-2338 mg/kg) of NaCl dissolved in physiological saline.
Details on exposure:
not applicable
Duration of treatment / exposure:
Dose-response relationships were studied in cells sampled 6 h after intraperitoneal administration of various amounts of NaCl. The time course of
aberrant cells was examined 3, 6, 12, 18 and 24 h after treatment with 40 mmole/kg (2338 mg/kg) of NaC1. The percentage of aberrant cells
reached a maximum level after 6 h.
Frequency of treatment:
not specified in the report
Post exposure period:
The time course of aberrant cells was examined 3, 6, 12, 18 and 24 h after treatment with 40 mmole/kg (2338 mg/kg)
of NaC1. The percentage of aberrant cells reached a maximum level after 6 h.
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
10 mmole/kg
Basis:
nominal conc.
Remarks:
Doses / Concentrations:
20 mmole/kg
Basis:
nominal conc.
Remarks:
Doses / Concentrations:
40 mmole/kg
Basis:
nominal conc.
No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
Positive control(s):
not specified in the report

Examinations

Tissues and cell types examined:
Bone marrow cells
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION: Not specified in the report.
TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): The time course of aberrant cells was examined 3, 6, 12, 18 and 24 h after treatment with 40 mmole/kg (2338 mg/kg) of NaC1. The percentage of aberrant cells reached a maximum level after 6 h with 40 mmole/kg (2338 mg/kg) of NaCl.

DETAILS OF SLIDE PREPARATION: Chromosome specimens were prepared from the femur bone marrow by the conventional
method (Sugiyama, 1971), stained in 2% Giemsa solution (pH 6.8) for 15 rain, and analyzed microscopically under blind code.

METHOD OF ANALYSIS: Chromosome aberrations (CA) defining aberrant metaphase cells consisted of breaks and gaps, and all breaks were of the chromatid type. Gaps were defined by a clear achromatic lesion with chromatid continuity, and breaks by discontinuity. Cells with gaps did not include in the category of damaged cells, based on the general opinion (Adler et al., 1971; Legator et al., 1973) that gaps are not a good indicator of chromosome damage. About 150-200 metaphase cells were examined per rat and the percent aberrant metaphase cells was calculated.
Each group consisted of 5 rats. Positive results were defined by significant differences in the X 2- test.

Evaluation criteria:
There are several criteria for determining a positive result, such as a dose-related increase in the relative number of cells with chromosome aberrations or a clear increase in the number of cells with aberrations in a single dose group at a single sampling time. Biological relevance of the results
should be considered first. Statistical methods may be used as an aid in evaluating the test results
.1) Statistical significance should not be the only determining factor for a positive response. Equivocal results should be clarified by further testing
preferably using a modification of experimental conditions.
2). An increase in polyploidy may indicate that the test substance has the potential to induce numerical chromosome aberrations. An increase in
endoreduplication may indicate that the test substance has the potential to inhibit cell cycle progression .
3). A test substance for which the results do not meet the above criteria is considered nonmutagenic in this test.
4). Although most experiments will give clearly positive or negative results, in rare cases the data set will preclude making a definite judgment about the activity of the test substance. Results may remain equivocal or questionable regardless of the number of experiments performed.
5). Positive results from the in vivo chromosome aberration test indicate that a substance induces chromosome aberrations in the bone marrow of
the species tested. Negative results indicate that, under the test conditions, the test substance does not induce chromosome aberrations in the bone
marrow of the species tested.
7). The likelihood that the test substance or its metabolites reach the general circulation or specifically the target tissue (e.g., systemic toxicity)
should be discussed.
Statistics:
Details on statistics not specified in the reports
Positive results were defined by significant differences in the X 2- test.

Results and discussion

Test results
Sex:
female
Genotoxicity:
positive
Toxicity:
not specified
Vehicle controls validity:
valid
Negative controls validity:
not specified
Positive controls validity:
not specified
Additional information on results:
The percentage of aberrant cells reached a maximum level after 6 h and thereafter decreased, but was not down to the control level after 24 h. Sucrose was tested to study whether NaCl-induced CA was due to high osmolarity. No significant result was noted at a dose of 30 mmole/kg
(10.269 g/kg). Sucrose was not soluble in physiological saline at doses greater than 30 mmole/kg.

Any other information on results incl. tables

Dose-response relationships were studied in cells sampled 6 h after intraperitoneal administration of various amounts of NaCl. Compared to the values for the untreated control, a significant difference was noted for the NaCl dose of 40 mmole/kg (2338 mg/kg). The incidence of aberrant

metaphase cells increased proportionally with the dose of NaCl. The time course of aberrant cells was examined 3, 6, 12, 18 and 24 h after treatment with 40 mmole/kg (2338 mg/kg) of NaCl. The percentage of aberrant cells reached a maximum level after 6 h and thereafter decreased,

but was not down to the control level after 24 h.

See the attached word in the attchment section doc ." Table for results" for the details.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): positive
A statistically significant positive result was obtained with 40 mmole/kg NaCl (2338 mg/kg).
Executive summary:

Sodium Chloride was evaluated for the chromosomal abberation test in the rat bone marrow cells. Female Long-Evans rats (a closed colony of

Long-Evans rats outbred from the colony of the Ben May Laboratory for Cancer Research, University of Chicago), aged 4-6 weeks and weighing

50-100 g were used. They were fed a commercial ration of NMF (Oriental Ferment Co., Tokyo) and water ad libitum, and kept at 22 ° C.

10-40 mmole/kg (584.4-2338 mg/kg) of NaCl dissolved in physiological saline.

Rat Bone marrow cells were examined for the chromosimal abberations. At various times after the intraperitoneal administration of the chemicals, the animals were killed for the analysis of chromosome aberrations in metaphase cells of the femur bone marrow cells. The animals were injected with colchicine (0.3 mg dissolved in 0.3 ml of physiological saline) 1 h before sacrifice. Chromosome specimens were prepared from the femur bone marrow by the conventional method (Sugiyama, 1971), stained in 2% Giemsa solution (pH 6.8) for 15 rain, and analyzed microscopically under blind code. Chromosome aberrations (CA) defining aberrant metaphase cells consisted of breaks and gaps, and all breaks were of the chromatid type.

gaps were not included in the category of damaged cells.

About 150-200 metaphase cells were examined per rat and the percent aberrant metaphase cells was calculated. Each group consisted of 5 rats. Positive results were defined by significant differences in the X 2- test.

Dose-response relationships were studied in cells sampled 6 h after intraperitoneal administration of various amounts of NaCl. Compared to

the values for the untreated control, a significant difference was noted for the NaCl dose of 40 mmole/kg (2338 mg/kg). The incidence of aberrant

metaphase cells increased proportionally with the dose of NaCl. The time course of aberrant cells was examined 3, 6, 12, 18 and 24 h

after treatment with 40 mmole/kg (2338 mg/kg) of NaCl.

The percentage of aberrant cells reached a maximum level after 6 h and thereafter decreased, but was not down to the control level after 24 h. Sucrose was tested to study whether NaCl-induced CA was due to high osmolarity. No significant result was noted at a dose of 30 mmole/kg

(10.269 g/kg). Sucrose was not soluble in physiological saline at doses greater than 30 mmole/kg.

Based on the results of the study the A statistically significant positive result was obtained with 40 mmole/kg NaC1 (2338 mg/kg).