A multitude of methods exists at present for the solubilization of biological tissues for atomic absorption analysis. We have examined several common methods of wet ashing using NBS bovine liver in order to determine which acids, acid combinations, or bases should be used as digesting agents for accurate and precise measurement of iron, copper, zinc, and manganese. Nitric acid proved to be the most effective wet ashing agent. With nitric acid, mean concentrations for iron, copper, and zinc differed from NBS certified values by less than 1.5% while those for manganese differed by 4%.

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The concentrations of iron, copper, zinc, manganese, calcium and magnesium, as well as protein, carbohydrate and fat, were analyzed in rat milk during the course of lactation in rats fed a purified diet. As serial milking has been demonstrated to affect milk composition (1), lactating rats were milked only once at various times from days 0 to 28 after parturition. Concentration of iron decreased rapidly from 9 to 5 micrograms/ml during the 1st week of lactation and changed little thereafter. Similarly, copper concentration decreased considerably in early lactation, from 9 to 2 micrograms/ml, but then remained constant. Zinc concentration did not decrease during the 1st week of lactation, but dropped from 14 micrograms/ml in the 2nd week of lactation to 8-10 micrograms/ml in mature milk. Manganese was unique as its concentration decreased during the early days of lactation, from 0.3 to 0.1 microgram/ml, with a subsequent rise in late lactation back to 0.3 microgram/ml. Calcium and magnesium concentrations increased somewhat in early lactation and decreased slightly in later lactation, but the changes were fairly small as compared to the trace elements. Developmental patterns for protein, carbohydrate and fat were not very pronounced, although protein and carbohydrate increased somewhat in early lactation and later decreased. These data demonstrate that the nutrient intake of the suckling changes markedly during the early neonatal period. These changes should be taken into account in evaluating data on the trace element composition of tissues of suckling rats.

Pregnant Swiss Webster mice were fed a diet moderately deficient in zinc from day 7 of gestation until parturition. Offspring of these mice showed depressed immune function through 6 months of age. In addition, the second and third filial generations, all of which were fed only the normal control diet, continued to manifest reduced immunocompetence, although not to the same degree as in the first generation.

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