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Analytical procedures are described for the determination of arsenic, cadmium, calcium, copper, lead, mercury, selenium, and zinc in animal feed. Mercury is determined by digesting the feed sample in a mixture of concentrated nitric and sulfuric acids with vanadium pentoxide added as an oxidation catalyst, reducing with stannous chloride, and sweeping the elemental mercury into an absorption tube for measurement by atomic absorption (AA) spectrophotometry. Arsenic and selenium are determined simultaneously by digesting the sample with a mixture of concentrated nitric, sulfuric, and perchloric acids; the hydrides of arsenic and selenium, which are formed with the addition of sodium borohydride, are swept into an argon-hydrogen flame for analysis by AA. A low temperature ash is prepared and dissolved in 1N HNO3 for the analysis of calcium, copper, and zinc by emission spectroscopy using the inductively coupled argon plasma source; the same solution is used for the determination of cadmium and lead by flameless AA. Animal feed spiked with 3 levels of each of the 8 elements gave recoveries that ranged from 80 to 107%.

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of each sex were assigned to each dose group fed either a cerealbased (NIH-31) or a purified (AIN-76A) diet, both containing nu-has been used successfully in managing the disease in patritionally adequate levels of copper. An additional control group tients who have developed this intolerance to penicillamine of rats and mice received a Cu-deficient AIN-76A diet. This low (Walshe, 1979(Walshe, , 1982 and is approved by the FDA for use copper diet resulted in Cu-deficiency symptoms, such as anemia, in these patients. The drug may be taken orally in maximum liver periportal cytomegaly, pancreatic atrophy and multifocal ne-daily adult dosages of 30 mg/kg, taken in two to four divided crosis, spleen hematopoietic cell proliferation, and increased heart portions. weight, together with undetectable levels of plasma copper in ratsFew studies of the toxicity of trien-2HCl have been rebut not in mice. Trien-2HCl lowered plasma copper levels someported in the peer-reviewed literature. In trying to determine what (at 600 and 3000 ppm) in rats fed the AIN-76A diet, but did optimal conditions for causing pancreatic acinar atrophy renot induce the usual signs of copper deficiency. Trien-2HCl caused an increased frequency of uterine dilatation at 3000 ppm in rats sulting from copper deficiency, Smith et al. (1982) incorpofed AIN-76A diet that was not noted in females fed the Cu-defi-rated triethylenetetramine tetrahydrochloride (trien-4HCl) cient diet. Trien-2HCl toxicity occurred only in mice in the highest into the diet of male Wistar rats at a concentration of 2000 dose group fed an AIN-76A diet. Increased frequencies of inflam-ppm. When the chelator was administered with standard rat mation of the lung interstitium and liver periportal fatty infiltra-chow for up to 8 weeks, minimal atrophy of the pancreas tion were seen in both sexes, and hematopoietic cell proliferation and minor changes in the lobular architecture of the tissue was seen in the spleen of males. Kidney and body weights were were noted. However, when administered with a Cu-defireduced in males as was the incidence of renal cytoplasmic vacuolicient diet for 6 weeks, pancreatic exocrine atrophy was very zation. There were no signs of copper deficiency in mice exposed marked, in excess of what was found with the Cu-deficient to trien-2HCl. The only effect of trien-2HCl in animals fed the NIH-31 diet was a reduced liver copper level in both rat sexes, diet alone. Other studies have shown trien-4HCl to be fetonoted at 3000 ppm. ᭧ 1996 Society of Toxicology toxic or teratogenic when fed to Sprague-Dawley rats during pregnancy at a level of 1660 ppm (Keen et al., 1983), and trien-2HCl was teratogenic to fetal mouse brain when Wilson's disease is a rare inherited metabolic disease char-administered during pregnancy at levels of 6000 or 12,000 acterized by an excessive accumulation of copper in certain ppm in the drinking water (Tanaka et al., 1993). Teratogenicity and effects on the exocrine pancreas possibly are second-1 To whom correspondence should be addressed....

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