39.1.28
Determination
AOAC Official Method 983.19
Calcium in Mechanically
Separated Poultry and Beef
Titrimetric Method
First Action 1983
Final Action 1985
A. Reagents
(a) Ethylenedinitrilotetraacetic acid, disodium salt, dihydrate
(EDTA).—0.02M. Dissolve 7.44 g EDTA (99+% purity) in H2O in 1 L
volumetric flask and dilute to volume. Standardize with ACS CaCO3
primary standard as in 967.30A(d) (see 18.4.07) (3 significant figures).
(b) Hydroxy naphthol blue indicator.—Mallinckrodt Specialty
Chemical Co.
(c) KOH–KCN solution.—Dissolve 280 g KOH in 500 mL H2O.
Cool, add 66 g KCN, dissolve, and dilute to 1 L in H2O. (Caution:
Use care in handling of KCN. HCN is evolved on contact with acid
or moisture.)
(d) Calcium carbonate.—0.02M. Weigh 2.000 g ACS primary
standard CaCO3 (dried 2 h at 100°C) and transfer to 1 L volumetric
flask. Add 500 mL H2O and 10–12 mL HCl (1 + 1). Heat just to boil-
ing to dissolve CaCO3. Cool and dilute to volume with H2O. Pipet
25 mL aliquot of EDTA (a) into 400 mL beaker and determine ratio
of EDTA to CaCO3 by titration as under Determination beginning
“…add ca 50 mL H2O, place on magnetic stirrer…”. Use average of
3 determinations.
B.
Weigh 10 g test sample, transfer to 300 mL tall form beaker, add
30 mL HCl (1 + 1), several glass beads, cover with watch glass, and
place on hot plate in fume hood. Bring to boil slowly, digest 20 min,
cool, and filter (Whatman No. 4 paper) into 200 mL volumetric
flask. Wash filter paper with H2O until 190 mL is obtained. Dilute to
volume and mix. Pipet 20 mL aliquot into 400 mL beaker, add ca
50 mL H2O, place on magnetic stirrer, and add 200–300 mg hydroxy
naphthol blue indicator, stir, and adjust pH to 12.5 ± 0.2 with
KOH–KCN solution. (Do not exceed pH 12.5 ± 0.2, to prevent pre-
cipitation. If precipitation occurs, discard and repeat.) Add
10–25 mL 0.02M EDTA accurately measured to provide
back-titration of minimum of 3 mL. Mix on magnetic stirrer and ti-
trate filtrate with 0.02M CaCO3 to permanent purple end point. If
back-titration is <3 mL, repeat with larger aliquot of EDTA solution.
Calcium (Ca), % = (A - B × R) × M × 4
Bone (for poultry), % = (%Ca – 0.015) × F
where A = mL EDTA added; B = mL CaCO3 of back titration; R =
EDTA:CaCO3 ratio; M = molarity of EDTA solution; 4 = meq.
weight Ca (0.040 g) × 100(%); 0.015 = correction for natural Ca in
poultry tissue; F = Ca-to-bone conversion factor (6.25 for young
chickens and 4.55 for turkeys and mature chickens).
Reference: JAOAC 66, 989(1983).
Revised: March 2002
© 2002 AOAC INTERNATIONAL