Inorganic Constituents

AACC Method 40-21 Page 1 of 2

Calcium in Feedstuffs
Final approval April 13, 1961; Reapproval November 3, 1999

Objective This method determines the calcium content of feedstuffs by a titrimetric procedure. Apparatus 1. Centrifuge. 2. Centrifuge tubes, tapered, 50-ml capacity. 3. Silica or porcelain ashing dish. 4. Muffle furnace, capable of operating at 600. 5. Acid-washed ashless filter paper. Reagents 1. Ammonium hydroxide (NH4OH), concentrated, ACS grade. 2. NH4OH (1 + 50). 3. HCl (1 + 1). 4. H2SO4, 1N. 5. Ammonium oxalate, saturated solution. Dissolve 4.2 g (NH4)2C2O4 in 100 ml water. 6. Dibromcresol purple indicator, 0.2% solution. 7. Potassium permanganate, 0.01N. Prepare and standardize as directed in Method 70-50A. Procedure 1. Weigh 2 g sample into silica or porcelain dish and ash at 600 for 17 hr. 2. After cooling, dissolve ash in 10 ml HCl (1 + 1), quantitatively transfer solution to 100-ml volumetric flask, and dilute to volume with water; mix thoroughly and, if cloudy, filter through acid-washed ashless filter paper. 3. Place aliquot (525 ml depending on Ca content) of solution in 50-ml tapering centrifuge tube, and dilute with water to 25-ml volume if necessary. Add 2 drops dibromcresol purple indicator and 10 ml saturated ammonium oxalate solution. Add concentrated NH4OH until lavender, add dilute HCl until pale yellow, and then adjust pH to 5.8 (dirty-gray color) by adding dilute NH4OH (1 + 50). Let stand overnight. 4. Centrifuge at 1300 rpm for 10 min. Decant supernatant liquid (save for Mg determination if desired), wash precipitate with NH4OH (1 + 50), and centrifuge and decant as before. 5. Dissolve calcium oxalate precipitate in tube with 10 ml 1 N H2SO4, place tube in boiling water bath, and titrate with 0.01N KMnO4 to faint pink end point.

Inorganic Constituents

AACC Method 40-21 Page 2 of 2

Calcium in Feedstuffs (continued)

Calculation If 10-ml aliquot is used in step 3:
% Ca = ml KMnO 4 N 10

where N = normality of KMnO4 solution. References

1. Fiske, C. H., and Subbarow, Y. 1925. Colorimetric determination of phosphorus. J. Biol. Chem. 66:375. 2. Kramer, B., and Howland, J. 1926. The quantitative estimation of calcium, magnesium, phosphate, and carbonate in bone. J. Biol. Chem. 68:711. 3. Morris, H. P., Nelson, J. W., and Palmer, L. S. 1931. Quantitative determination of calcium, magnesium, and phosphorus in feedstuffs and cattle excreta (improved technic). Ind. Eng. Chem., Anal. Ed. 3:164.