Inorganic Constituents

AACC Method 40-20 Page 1 of 2

Calcium in Bread and Flour

Final approval May 5, 1960; Reapproval November 3, 1999

Objective This method determines the calcium content of flour and bread by a titrimetric procedure. Apparatus 1. Platinum, silica, or porcelain ashing dishes, about 60-mm diameter, 35-ml capacity. Porcelain evaporating dishes of about 25-ml capacity are satisfactory. Do not use flat-bottom dishes of greater diameter than 60 mm. 2. Muffle furnace capable of operating at 550. 3. Filter (choose one of the following): a. Quantitative filter paper, medium speed; Whatman No. 40 or equivalent. b. Fritted glass crucible, fine porosity. c. Microporous porcelain filter crucible, medium porosity. 4. Glassware as required. 5. Acid-washed ashless filter paper. Reagents 1. Ashing aid. a. Magnesium nitrate solution. Dissolve 50 g Mg(NO3)26H2O in water and dilute to 100 ml. Or: b. Redistilled HNO3. 2. Oxalic acid solution, 3%. 3. Potassium permanganate, 0.05N. Prepare and standardize as directed in Method 70-50A. 4. Bromcresol green indicator, 1%. 5. Sodium acetate solution, 20%. 6. Ammonium hydroxide (1 + 50). 7. Hydrochloric acid (HCl), concentrated, ACS grade. 8. Sulfuric acid (H2SO4), concentrated, ACS grade. Procedure 1. Weigh 10.0 g prepared sample (for bread, Method 62-05) into ashing dish that has been ignited, cooled in desiccator, and weighed soon after attaining room temperature. Begin preparation of reagent blank at this point. 2. Incinerate in furnace at about 550 (dull red) until light-gray ash results, or to constant weight. See Note. 3. Cool in desiccator and weigh soon after room temperature is attained, if percent ash is desired. 4. Add 5 ml HCl, allowing acid to rinse upper portion of dish; evaporate to dryness on steam bath.

Inorganic Constituents

AACC Method 40-20 Page 2 of 2

Calcium in Bread and Flour (continued)

5. Dissolve residue by adding, accurately measured, 2.0 ml HCl; heat 5 min on steam bath with watch glass on dish. 6. Wash off watch glass with water; then filter through fast acid-washed ashless filter paper into 400-ml beaker. Dilute to about 150 ml. 7. Add 810 drops bromcresol green indicator and sufficient 20% sodium acetate solution to change pH to 4.85.0 (blue). Cover with watch glass and heat to boiling. 8. Precipitate calcium slowly by adding 3% oxalic acid solution, 1 drop every 35 sec, until pH is changed back to 4.44.6 (optimum for calcium oxalate precipitation) as indicated by appearance of distinct green shade. (Change of color indicates excess of oxalic acidmore would develop yellow tints, showing undesirable displacement of pH.) 9. Boil 12 min and let mixture settle until clear, or overnight. 10. Filter supernatant liquid through quantitative filter paper, fritted glass filter, or microporous crucible. 11. Rinse beaker and transfer precipitate to filter with about 50 ml NH4OH (1 + 50) in small portions, using wash-bottle delivering very small stream. Discard filtrate. 12. Dissolve precipitate by washing filter or crucible with mixture of 125 ml water and 5 ml H2SO4 at 8090. 13. Titrate filtrate at 7090 with 0.05N KMnO4 until slight pink color is obtained, add filter paper or filter, and continue titration if necessary. 14. Correct for blank. Calculation Calculate calcium as mg/lb; 1 ml 0.05N KMnO4 = 1 mg calcium. Note To diminish ashing time, or for samples that do not burn practically carbonfree, use one of the following ashing aids: Moisten ash with a) 0.51.0 ml magnesium nitrate solution or b) redistilled HNO3. Dry contents and carefully ignite in muffle to prevent spattering. (A white ash with no carbon results in most cases.) Do not add these ashing aids to self-rising flour (products containing NaCl) in platinum dish, because of vigorous action on dish. Reference
AOAC International. 1995. Official Methods of Analysis of AOAC International, 16th ed. Methods 944.03 and 945.41. The Association, Arlington, VA.