Eight commercially available organic Zn products and reagent-grade ZnSO4 x 7H2O (Zn Sulf) were evaluated by polarographic analysis, and solubility in .1 M K2HPO4-KH2PO4 buffer (pH 5), .2 M HCl-KCl buffer (pH 2), and deionized water. Fractions from these solubility tests were evaluated by gel filtration chromatography for structural integrity. Degree of chelation was generally positively related to chelation effectiveness determined by polarography. The organic sources were Zn methionine complex A (Zn MetA), Zn methionine complex B (Zn MetB), Zn polysaccharide complex (Zn Poly), Zn lysine complex (Zn Lys), Zn amino acid chelate (Zn AA), Zn proteinate A (Zn ProA), Zn proteinate B (Zn ProB), and Zn proteinate C (Zn ProC). Three experiments were conducted to estimate the relative bioavailability of Zn from the organic Zn supplements for chicks and lambs when added at high dietary levels to practical diets. Bone Zn concentration increased (P < .001) as dietary Zn increased in both experiments. When Zn Sulf was assigned a value of 100% as the standard, multiple linear regression slope ratios of bone Zn from chicks fed 3 wk regressed on dietary Zn intake gave estimated relative bioavailability values of 83 +/- 14.6 and 139 +/- 16.9 for Zn AA and Zn ProA, respectively, in Exp. 1 and 94 +/- 11.6, 99 +/- 8.8, and 108 +/- 11.4 for Zn Poly, Zn ProB, and Zn ProC, respectively, in Exp. 2. In Exp. 3, 42 lambs were fed diets containing Zn Sulf, Zn ProA, Zn AA, or Zn MetB for 21 d. Based on multiple linear regression slope ratios of liver, kidney, and pancreas Zn and liver metallothionein concentrations on added dietary Zn, bioavailability estimates relative to 100% for Zn Sulf were 130, 110, and 113 for Zn ProA, Zn AA, and Zn MetB, respectively. Except for Zn ProA, which was greater, the organic Zn supplements had bioavailability values similar to that of Zn Sulf for chicks and lambs. Bioavailability of organic Zn products was inversely related to solubility of Zn in pH 5 buffer in chicks (r2 = .91) and pH 2 buffer in lambs (r2 = .91), but not to an estimate of degree of chelation.
Five commercially available organic Cu products and reagent-grade CuSO4 x 5H2O (Cu Sulf) were evaluated by polarographic analysis and solubility in 0.1 M K2HPO4-KH2PO4 buffer (pH 5), 0.2 M HCl-KCl buffer (pH 2), or deionized water. Fractions from these solubility tests were evaluated by gel filtration chromatography for structural integrity. The organic sources were Cu lysine complex (Cu Lys), Cu amino acid chelate (Cu AA), Cu proteinate A (Cu ProA), Cu proteinate B (Cu ProB), and Cu proteinate C (Cu ProC). Separation of peaks in the chromatograms for the soluble Cu fraction from deionized water indicated that 77, 31, 69, 94, and 16% of the Cu remained chelated for the above sources, respectively. Two experiments were conducted to estimate the relative bioavailability of Cu from the organic Cu supplements for chicks when added at high dietary concentrations to practical corn-soybean meal diets. Liver Cu concentration increased (P < 0.0001) as dietary Cu increased in both experiments. When Cu Sulf was assigned a value of 100% as the standard, linear regression slope ratios of log10 liver Cu concentration regressed on added dietary Cu concentration gave estimated relative bioavailability values of 124 +/- 5.1, 122 +/- 5.3, and 111 +/- 6.0 for Cu Lys, Cu AA, and Cu ProC, respectively, in Exp. 1. The bioavailability estimates for Cu Lys and Cu AA were greater (P < 0.05) than that for Cu Sulf. Values in Exp. 2 were 111 +/- 7.6, 109 +/- 8.4, and 105 +/- 7.5 for Cu Lys, Cu ProA, and Cu ProB, respectively, and all sources were similar in value for chicks. Solubility of Cu in pH 2 buffer provided the best prediction of bioavailability (r2 = 0.924). Other indicators of chelation integrity and solubility had little value as predictors of bioavailability (r2 < or = 0.445).
A specific, sensitive and reliable index for assessment of human zinc status has not been developed, and continues to present a considerable challenge for nutritionists in the trace element field. We have focused on metallothionein (MT) expression as a potential index. A protocol involving 16 men and a 10-d supplementation period plus a 4-d postsupplementation period was used to examine the relative response of MT expression in erythrocytes, monocytes, peripheral blood mononuclear cells (PBMC) and cells from a dried blood spot (DBS). Zinc was supplemented at the current adult male recommended dietary allowance (RDA) of 15 mg. Erythrocyte MT protein, as measured by ELISA, increased gradually to about twofold over the placebo group during zinc supplementation and remained elevated for 4 d postsupplementation. Competitive reverse transcriptase-polymerase chain reaction showed that MT mRNA levels in both monocytes and PBMC increased (up to 4.7- and 2.7-fold, respectively) after 2 d of supplementation, with greater expression in monocytes compared with PBMC. Total RNA extracted from dried blood spots, representing cells from 50 microL of blood, showed a comparable change in MT mRNA upon zinc supplementation. In each leukocyte population isolated, when zinc supplementation was withdrawn, MT mRNA levels decreased. Collectively, these experiments show that, in men, MT gene expression increases during supplementation at the RDA, and that the DBS sampling method will be of value in measuring MT expression in a variety of clinical and survey situations.
Three experiments were conducted with day-old chicks to study the effects of dietary Fe concentration and age on Fe accumulation in tissues as an estimate of supplemental Fe bioavailability, and of delaying the time of initial high Fe supplementation up to 7 d of age on feed intake to 3 wk of age. In Experiment 1, chicks were fed a basal corn-soybean meal diet (188 mg/kg Fe, DM basis) or the basal supplemented with 400, 600, or 800 mg/kg added Fe as reagent grade FeSO4.7H2O for either 1, 2, or 3 wk. Dietary Fe depressed (P < 0.001) feed intake and body weight gain, especially at 3 wk. Kidney Fe concentrations increased linearly (P < 0.001) with increasing dietary Fe. Liver Fe concentration also increased linearly, but reached a plateau in birds fed 600 mg/kg Fe. Bone Fe increased linearly (P < 0.05) at 1 wk, but not at 2 or 3 wk. Liver and kidney Fe regressed on daily Fe intake had the best fit to a linear model at 2 wk. In Experiment 2, chicks were fed either a basal diet (320 mg/kg Fe, DM basis) continuously, the basal supplemented with 800 mg/kg added Fe as FeSO4.7H2O continuously, or were started on the control diet and switched to the high Fe diet on Day 3, 5, or 7. Feed intake was lower (P < 0.05) in birds started on Fe on Days 1 or 3, but delaying feeding of high Fe diets until Day 5 resulted in intake at 3 wk similar to that of birds fed the basal diet. In Experiment 3, the basal diet (123 mg/kg Fe) was fed to chicks for 6 d, then experimental diets were fed for 14 d. Diets were the basal or basal supplemented with 400, 600, or 800 mg/kg added Fe as reagent grade or feed grade Fe sulfate or an Fe methionine complex. When estimated from regression of log10 liver Fe concentration on total analyzed dietary Fe concentration, relative bioavailability was set at 100% for reagent grade Fe sulfate, and the feed grade sulfate was 92.3% and Fe methionine was 88.3%. Liver Fe concentrations may be useful criteria for determining Fe bioavailability and 2 wk of feeding was the optimal time required for such a bioassay. Delaying feeding high Fe diets until 5 d of age alleviated the decreased feed intake associated with high Fe diets.
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