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Toxicological information

Genetic toxicity: in vivo

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Administrative data

Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
1990-04-27 to 1990-09-21
Reliability:
2 (reliable with restrictions)
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1990
Report Date:
1990

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Principles of method if other than guideline:
The purpose of this study is to evaluate the potential of the test article to increase the incidence of micronucleated polychromatic erythrocytes in bone marrow of male and female mice.
In vivo cytogenetic analysis provides a valuable tool for the detection of chromosome breakage. Micronuclei usually arise from acentric chromosome fragments or whole chromosomes which are lost from the nucleus during replication of erythrocyte precursor cells and remain in the cytoplasm of young, anucleate (polychromatic) erythrocytes after expulsion of the nucleus. The increase in incidence of micronucleated cells has two primary causes, chromosome breakage and malfunction of the spindle apparatus.
The clastogenic potential of the test article is measured by its ability to significantly increase the incidence of micronucleated polychromatic erythrocytes when compared with control animals.
see also:
Heddle, J.A., et al.; Mut. Res. 123, pp 61 - 118, 1983;
Matter, B.E., Grauwiler, J.; Mut. Res. 23, pp 239 - 249, 1973.
GLP compliance:
yes
Type of assay:
micronucleus assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: sodium persulfate
- Molecular formula: H2Na2O8S2
- Physical state: white solid
- Analytical purity: > 99 %
- Lot/batch No.: E608244 Lot 0021
- Expiration date of the lot/batch: not stated
- Stability under test conditions: stable at room temperature
- Storage condition of test material: Dry conditions; at room temperature, in the dark.

Test animals

Species:
mouse
Strain:
ICR
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Sprague Dawley Inc., Frederick, MD or Charles River Breeding Laboratories, Kingston, NY or Raleigh, NC
- Age at study initiation: 6 - 8 weeks
- Weight at study initiation: Toxicity studies: Males 24 - 34 grams; Females: 21 - 25 grams; Micronucleus assay: Males: 27 - 35 grams; Females. 21 - 27 g; all weights measured at dose administration
- Assigned to test groups randomly: yes
- Fasting period before study: not indicated
- Housing: five per cage in autoclavable plastic cages with hardwood chips as bedding material
- Diet: certified laboratory rodent chow ad libitum
- Water: tap water ad libitum
- Acclimation period: not less than 5 days;


ENVIRONMENTAL CONDITIONS
- Temperature: 20 - 26.7 °C
- Humidity: 50 +/- 20 % R.H
- Air changes: not indicated
- Photoperiod: 12 hrs dark / 12 hrs light;

Administration / exposure

Route of administration:
intraperitoneal
Vehicle:
Distilled water;
Details on exposure:
For the initial toxicity study, animals were randomly assigned to five groups of five males and five females each. All animals were weighed immediately prior to dose administration.
For the micronucleus assay, the animals were assigned to thirteen experimental groups of five males and five females, each based on a computer-generated randomization program. An additional group of five males and five females were designated as replacement animals in the event of mortality prior to the scheduled sacrifice time and were dosed with the test article high dose level. Each animal was given a sequential number and identified by ear tag.
The test article-vehicle mixture or the vehicle alone were administered by IP injection at a constant rate of 10 mL/kg bw. The positive control, TEM, was injected IP at a dose of level of 0.25 mg/kg. All mice in the experimental and control groups were weighed immediately prior to dose administration and the dose volumes based on individual body weights . Animals were observed after dose administration for clinical signs of chemical effect.
Duration of treatment / exposure:
Scheduled sacrifice time after administration of the test article:24; 48; 72 h
Frequency of treatment:
One treatment
Post exposure period:
24; 48; 72 h
Doses / concentrations
Remarks:
Doses / Concentrations:
85; 169; 338 mg/kg bw
Basis:
nominal conc.
No. of animals per sex per dose:
5 animals per sex per dose
Control animals:
yes, concurrent vehicle
Positive control(s):
yes; TEM - Triethylene melamine; 0.25 mg/kg bw

Examinations

Tissues and cell types examined:
Bone marrow of the femur
Details of tissue and slide preparation:
At the scheduled sacrifice time, five mice per sex were sacrificed by CO2 asphyxiation. Immediately following sacrifice, the femurs were exposed, cut just above the knee, and the bone marrow was aspirated into a syringe containing foetal bovine serum. The bone marrow cells were transferred to a capped centrifuge tube containing approximately 1 mL FBS. The bone marrow cells were pelleted by centrifugation at approximately 100 x g for five minutes and the supernatant was drawn off, leaving a small amount of serum with the remaining cell pellet. The cells were resuspended by aspiration with a capillary pipet and a small drop of bone marrow suspension was spread onto a clean glass slide. Two to four slides were prepared from each animal. The slides were fixed in methanol, stained with May-Gruenwald-Giemsa and permanently mounted.
Slides were coded using a random number table by an individual not involved with the scoring process. Using medium magnification, an area of acceptable quality was selected such that the cells were well spread and stained. Using oil immersion, 1000 polychromatic erythrocytes were scored for the presence of micronuclei. The number of micronucleated normocytes in the field of 1000 polychromatic erythrocytes was enumerated. The proportion of polychromatic erythrocytes to total erythrocytes was also recorded.
Evaluation criteria:
The test article is considered to induce a positive response if a treatment-related increase in micronucleated polychromatic erythrocytes is observed relative to the vehicle control (p <= 0.05, Kastenbaum-Bowman tables). The positive response must be dose-dependent or must be observed at a single dose level at adjacent sacrifice times. If a single treatment group is significantly elevated at one sacrifice time, the assay is considered a suspect or unconfirmed positive and a repeat assay will be recommended.
Statistics:
Statistical significance will be determined using the Kastenbaum-Bowman tables which are based on binomial distributions.

Results and discussion

Test results
Sex:
male/female
Genotoxicity:
negative
Toxicity:
yes
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
For the initial toxicity study, sodium persulfate was administered by IP injection to male and female ICR mice at five treatment levels and a vehicle control. 208, 292, 408, 571 or 800 mg test article/kg body weight which was administered in a total volume of 10 mL test article-vehicle mixture/kg body weight. Mortality which occurred shortly after dose administration and up to four days later, was 0/10 at 208 and 292 mg/kg, 6/10 at 408 mg/kg and 10/10 at 571 and 800 mg/kg. The LD50 was calculated by probit analysis to be approximately 422 mg/kg. The high dose for the micronucleus test was set at 338 mg/kg which was estimated to be 80 % of the LD50.

Any other information on results incl. tables

TABLE :MICRONUCLEATED POLYCHROMATIC ERYTHROCYTES IN BONE MARROW: SUMMARY 

TREATMENT

SEX

TIME

(HR)

NUMBER OF

MICE

PCE/TOTAL

ERYTHROCYTES

MICRONUCLEATED POLYCHROMATIC ERYTHROCYTES

NUMBER PER

1000 PCE’S

NUMBER PER

PCE'S SCORED

(MEAN

±

S.D.)

Water

 

 

 

 

 

 

 

 

10 ml/kg

M

24

5

0.57

0.6

±

0.89

3 / 5000

 

 

48

5

0.48

1.6

±

0.89

8 / 5000

 

 

72

5

0.57

0.4

±

0.89

2 / 5000

 

F

24

5

0.62

1.2

±

0.84

6 / 5000

 

 

48

5

0.53

1.2

±

0.84

6 / 5000

 

 

72

5

0.54

1.0

±

1.00

5 / 5000

Sodium persulfate, E608244, Lot 0021

85 mg/kg

M

24

5

0.60

0.6

±

0.55

3 / 5000

 

 

48

5

0.48

1.6

±

0.55

8 / 5000

 

 

72

5

0.50

1.0

±

1.00

5 / 5000

 

F

24

5

0.65

1.4

±

1.14

7 / 5000

 

 

48

5

0.59

1.8

±

0.84

9 / 5000

 

 

72

5

0.60

0.8

±

1.10

4 / 5000

169 mg/kg

M

24

5

0.60

1.4

±

0.89

7 / 5000

 

 

48

5

0.59

0.8

±

0.45

4 / 5000

 

 

72

5

0.64

1.0

±

1.00

5 / 5000

 

24

5

0.63

0.8

±

0.84

4 / 5000

 

 

48

5

0.65

1.2

±

0.45

6 / 5000

 

 

72

5

0.45

1.2

±

1.10

6 / 5000

338 mg/kg

M

24

5

0.51

0.2

±

0.45

1 / 5000

 

 

48

5

0.54

1.0

±

0,71

5 / 5000

 

 

72

4

0.47

1.0

±

0.82

4 / 4000

 

F

24

5

0.56

1.0

±

0.71

5 / 5000

 

 

48

5

0.57

2.4

±

1.52

12 / 5000

 

 

72

3

0.37

2.7

±

2.08

8 / 3000

TEM

 

 

 

 

 

 

 

 

0.25mg/kg

M

24

5

0.55

59.8

±

22.42

299 / 5000*

 

F

24

5

0.54

49.6

±

8.38

248 / 5000*

1*, p <=0.05 (Kastenbaum-Bowman Tables)

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative
No significant increase in micronucleated polychromatic erythrocytes was observed at 24, 48 or 72 hours after dose administration in male or female ICR mice. Thus, disodium persulfate was considered to be not clastogenic.
Executive summary:

In the presented mouse micronucleus assay, male and female ICR mice were exposed to 85, 169 or 338 mg/kg of disodium persulfate which was administered at a constant rate of 10 mL/kg as a single IP injection. The high dose level was calculated to be 80 % of the LD50. Bone marrow cells, collected 24, 48 and 72 hours after treatment and were examined microscopically for micronucleated polychromatic erythrocytes. A reduction in the ratio of polychromatic erythrocytes to total erythrocytes was observed in female mice at 72 hours after administration of 169 mg/kg and in male and female mice at 72 hours after administration of 338 mg/kg, indicated that the test substance did induce bone marrow toxicity. No significant increases in micronucleated polychromatic erythrocytes was observed at 24, 48 or 72 hours after dose administration in males or females. The results of the assay indicated that under the conditions described disodium persulfate did not induce a significant increase in micronucleated polychromatic erythrocytes in male or female ICR mice. Disodium persulfate was concluded to be negative in the mouse micronucleus assay. Thus, disodium persulfate was considered to be not clastogenic.