AMMONIUM SULFATE CAS N°: 7783-20-2

OECD SIDS
AMMONIUM SULFATE
5. TOXICITY ID: 7783-20-2
DATE: 18.04.2006
Alu I + 0.8M (NH4)2SO4 / 59.0 / 22.0
Alu I + 1.6M (NH4)2SO4 / 76.5 / 42.5
Alu I + 3.2M (NH4)2SO4 / 98.2 / 240.3
control (Alu buffer) / 16.1 / 4.1
Alu buffer + 3.2 M (NH4)SO4 / 22.5 / 6.0
Test condition:
*DIC: Percent aberrant metaphases, including achromatic
lesions and dicentric chromosomes per 100 metaphases as
calculated from all polycentric chromosomes induced in CHO
cells by 24 units Alu I in the presence or absence of salts.
Four possibilities for this effect were discussed by the
authors: (1) salt enhances the permeability of the cell
membrane, (2) salt changes the structure of the chromatin,
(3) salt changes the structure of the DNA, (4) salt
influences the repair of damaged DNA.
Pellets of 4 x 10e6 cells were made from cultures grown for
2 days and washed once with 1 mL newborn calf serum (NCS).
The pellets received 24 units Alu I (Boehringer Mannheim) in a
volume of 8 µl containing Alu I (3.4 or 4 µL) and NCS.
After mixing, the cells were incubated for 20 minutes at 37
°C in the incubator. Ammonium sulfate was dissolved in
distilled water and 10 µL were added 5 minutes after the
addition of the AluI. The cells were mixed and reincubated for
a further 15 minutes. Respective controls were set up in the
same way, but instead of Alu I the cells were treated with the
buffer in which the Alu I was shipped.
After the treatment the cells of each pellet were seeded in
2 6-cm petri dishes and incubated in medium containing fetal
calf serum instead of NCS (10%), and bromodeoxyuridine
(BrdUrd) in a final concentration of 2 x 10e-5 M.
Preparations were made 18 hours later including a 2-hour
treatment with Colcemid. The chromosomes were differentially
stained following the method of Hill and Wolff (1982, Cancer
Res. 42, 893) and exclusively first post-treatment metaphases
were analyzed with respect to chromosomal aberrations. The
number of dicentric chromosomes (DIC) per 100 metaphases were
calculated from all polycentric chromosomes in such a way that
from the polycentric chromosomes with more than 2 centromeres,
one centromere was subtracted and the remaining number of
centromeres was taken as the number of DIC.
Number of metaphases analyzed / number of independent
experiments:
Alu I: 1100 / 11
Alu I + 0.8M ammonium sulfate: 200 / 2
Alu I + 1.6M ammonium sulfate: 200 / 2
Alu I + 3.2M ammonium sulfate: 600 / 6
Alu buffer (control): 1100 / 11
Alu buffer + 3.2M ammonium sulfate: 200 / 2
Year of study: not reported.
Reliability: (2) valid with restrictions
limited documentation, no information on purity, no standard
chromosome aberration test
Flag:
Critical study for SIDS endpoint
12-APR-2006 (141)
Type:
System of testing:
188
Cytogenetic assay
CHO cells
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