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EC number: 215-572-9
CAS number: 1332-65-6
Materials and Methods:
The aim of the study was to
examine the effect of copper sulphate (0, 500, 1000, 2000, 4000
or 8000 ppm) administered to male and female B6C3F1mice
in feed for 13 weeks. The test organisms were observed throughout the
study for signs of clinical toxicity, mortality, bodyweight changes and
food consumption. Throughout the study blood and urine samples were
collected to determine haematology, clinical chemistry and urinalysis
parameters and tissue metal level. At
the end of the study period all animals were sacrificed and subject to
pathological examinations to determine any histological, sperm
morphology or vaginal cytology abnormalities. The
study was conducted to a methodology developed by the US National
Toxicology Programme specifically for the test. The study was conducted
in accordance with GLP.
Results and Discussion
chemistry and urinalysis evaluations of rats revealed variable
chemical-related changes that were, for the most part, restricted to the
4000 and 8000 ppm groups. Increases in serum alanine aminotransferase
and sorbitol dehydrogenase activities in both sexes were indicative of
hepatocellular damage, as were increases in 5’-nucleotidase and bile
salts in males. Decreases in mean cell volume, hematocrit and
haemoglobin indicated the development of a microcytic anaemia, while
increases in reticulocyte numbers at the same time points suggested a
compensatory response to the anaemia by the bone marrow. Increases in
urinary glucose and N-acetyl-β-D-glucosaminidase (a lysosome enzyme) and
asparate aminotransferase (a cytosolic enzyme) were suggestive of renal
tubule epithelial damage.
Dose related increases in copper
occurred in all male rat tissues examined. These increases were
accompanied by increases in zinc in the liver and kidney. Plasma
calcium was significantly reduced in the 4000 and 8000 ppm groups, and
there was a trend towards reduction in calcium in the kidney and testis
as well. In the 8000 ppm group, plasma magnesium was significantly
increased relative to the controls.
Rats in the three highest dose
groups had hyperplasia and hyperkeratosis of the forestomach,
inflammation of the liver and increases in the number and size of
protein droplets in the epithelial cytoplasm and the lumina of the
proximal convoluted tubules. Many of the droplets in the male kidneys
were large and had irregular crystalline shapes. These droplets stained
strongly positive for protein but were negative for iron, PAS, and
acid-fast (lipofuscin) staining methods. Α-2-microglobulin was present
in the droplets of male rats, but there was no dose-related qualitative
difference in the content of this protein. In the 4000 and 8000 ppm
groups, copper was distributed in a periportal to midzonal pattern in
the liver and was restricted to the cytoplasm of the proximal convoluted
tubule epithelium in the kidney. Copper was present in some, but not
all, of the protein droplets. Transmission electron microscopy of the
livers of rats of each sex revealed increases in the number of secondary
lysosomes in hepatocytes in the periportal area.
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