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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:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: OECD protocl study under GLP; no deviations

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1991
Report date:
1991

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
GLP compliance:
yes (incl. QA statement)
Type of assay:
micronucleus assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Sodium 2-methylprop-2-ene-1-sulphonate
EC Number:
216-341-5
EC Name:
Sodium 2-methylprop-2-ene-1-sulphonate
Cas Number:
1561-92-8
Molecular formula:
C4H8O3S.Na
IUPAC Name:
sodium 2-methylprop-2-ene-1-sulfonate
Test material form:
solid: crystalline
Details on test material:
- Name of test material (as cited in study report): natriummethallylsulfonat
- Substance type: organic
- Physical state: solid crystalline
- Analytical purity: 99.8 %
- Impurities (identity and concentrations):
- Composition of test material, percentage of components: < 0.2% water. 0.05% naCl
- Isomers composition:
- Purity test date: Oct 1989
- Lot/batch No.: unknown
- Expiration date of the lot/batch: undeterminate
- Stability under test conditions: stable
- Storage condition of test material: Normal lab conditions

Test animals

Species:
mouse
Strain:
other: BOR:NMRI (SPF Harlan)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Fa Winkelmann, 4799 Borchem , Germany
- Age at study initiation: young adult mice
- Weight at study initiation: male 23.3 +- 0.9 ; female 23.5 +- 0.7
- Assigned to test groups randomly: [no/yes, under following basis: ] yes
- Fasting period before study: 18 hrs
- Housing: sex-separated, 5 anmials / Makrolon cage type III
- Diet (e.g. ad libitum): Saniff R10, Fa. Sniff 4770, Soest
- Water (e.g. ad libitum): ad lib
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 C +-1
- Humidity (%): 60 %+- 10
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: within lab: from 19-04-1990 to 28-09-1990

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
aqua bidest
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

Duration of treatment / exposure:
24, 48 and 72 hrs
Frequency of treatment:
single dose
Post exposure period:
None
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
1250 mg/kg bw
Basis:
actual ingested
dose determined on the basis of body weight immediately prior to dosing
Remarks:
Doses / Concentrations:
2500 mg/kg bw
Basis:
actual ingested
dose determined on the basis of body weight immediately prior to dosing
Remarks:
Doses / Concentrations:
5000 mg/kg bw
Basis:
actual ingested
dose determined on the basis of body weight immediately prior to dosing
No. of animals per sex per dose:
5 / 5
Control animals:
yes
yes, concurrent vehicle
Positive control(s):
cyclophosphamide;
- Justification for choice of positive control(s): Known positive, historical data available
- Route of administration: oral gavage
- Doses / concentrations: 100 mg/kg bw

Examinations

Tissues and cell types examined:
1000 polychromatic erythrocytes / animal
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION: dose-range finding with 5000, 6500, 7500 and 10000 mg/kg bw established confirmed highest LD (zero) of 5000 mg/kg bw


TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields):
after preparation of both femurs, the bone marrow is washed out using fetal bovine serum and a syringe. the cell suspension was centrifuged for 5 minutes at 160 g, then decanted and re-suspended. This suspension was cleaned by means of a cellulose chromatography tube.
The eluate from the tube was centrifuged (10 min/800 g) , the pellet was re-suspended in fetal bovine serum with 25mM EDTA.
DETAILS OF SLIDE PREPARATION:
From this suspension, 3-4 slides were prepared, dried for 24 hrs and colored with May-Gruenwald / Giemsa solution

METHOD OF ANALYSIS:
100-fold enlargement, Zeiss Microscope
1000 PCE cells/. animals
number of PCE cells with micronuclei
PCE /NCE determined
number of NCE cells with micronuclei .

Evaluation criteria:
Positive if a statistically significant increase in the frequency of micronucleus-containing PCE is found in at least one test group relatie to the neagative controls
Statistics:
Not specified

Results and discussion

Test results
Sex:
male
Genotoxicity:
negative
Toxicity:
yes
Remarks:
discolored feces (all animals and mild diarrhea in males
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
RESULTS OF RANGE-FINDING STUDY
- Dose range: 500, 6500, 7500, 10000 mg/kg bw
- Solubility: 750 g/l
- Clinical signs of toxicity in test animals: sedation and 1/3 fatality in 6500 group, sedation and 1 / 5 fatalties in teh 7500 group, sedation and 3/ /5 fatalities in the 10000 group.
- Harvest times: 24, 48, 72 hrs

RESULTS OF DEFINITIVE STUDY


- Induction of micronuclei (for Micronucleus assay): see table
- Ratio of PCE/NCE (for Micronucleus assay): see tabel
- Appropriateness of dose levels and route: route is appropriate in view of exposure potential; dose: you can't go much higher, can you?.
- Statistical evaluation: see table.

Any other information on results incl. tables

Testgruppe

Dosis

Entnahme nach

% PCE mit

PCE/NCE

 

(mg/kg KG)

Appl. (in h)

MK (x ± SD)

(x ± SD)

Positivkontrolle

 

 

 

 

(Cyclophosphamid)

100

24

* 3,95 ± 1,37

* 0,48 ± 0,2

 

_

24

0,07 ± 0,05

1,13 ± 0,2

Negativkontrolle

 

 

 

 

(Wasser)

-

48

0,25 ± 0,12

1,31 ± 0,09

 

-

72

0,28 ± 0,10

1,20 ± 0,33

 

5000

24

0,09 ± 0,06

0,97 ± 0,2

NATRIUM­

 

 

 

 

METHALLYL­

5000

48

0,18 ± 0,08

1,26 ± 0,16

SULFONAT

 

 

 

 

(Gruppe I)

5000

72

0,18 ± 0,11

1,38 ± 0,20

MAS-Na (Gruppe II)

2500

24

0,13 ± 0,06

1,10 ± 0,3

MAS-Na (Gruppe III)

1250

24

0,05 ± 0,05

1,20 ± 0,2

* statistisch signifikante Veranderung gegenüber der Negativkontrolle (Vertrauensbereich = 95 %)


Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative No significant increase in frequency of miconuclei
Substance is not clastogenic in a mouse micronucleas test at dose levels of 1250, 2500 and 5000 mg/kg bw.
Executive summary:

Substance was tested in a mouse bone marrow micronucleus test according to OECD 474.

Substance is not clastogenic in this test at dose levels of 1250, 2500 and 5000 mg/kg bw.