9.2.

38
AOAC Official Method 969.32
Zinc in Food
Atomic Absorption Spectrophotometric Method
First Action 1969
Final Action 1971
A. Principle
Material sample is dry or wet ashed. Residue is taken up in acid
and diluted to optimum working range. A of this solution as deter-
mined by AAspectrophotometry at 213.8 nmis converted to Zn con-
centration through calibration curve.
B. Reagents
(Use Pyrex glassware exclusively; clean thoroughly before use
with hot HNO
3
. If glass beads are used to prevent bumping, clean
first with strong alkali followed by hot HNO
3
. Since Pt used in labo-
ratory may contain significant traces of metals, clean Pt dishes by
KHSO
4
fusion followed by 10% HCl leach.)
(a) Zinc standard solutions.(1) Stock solution.500 g/mL.
Dissolve 0.500 g pure Zn metal in 510 mL HCl. Evaporate almost
to dryness and dilute to 1 Lwith H
2
O. Solution is stable indefinitely.
(2) Working solution.Dilute aliquots of stock solution with H
2
SO
4
(1 + 49) or 0.1M HCl (depending on method of ashing) to obtain
5 solutions within range of instrument. Prepare standards in
010g/mL range daily. (Do not use <2 mL pipets or <25 mL volu-
metric flasks.)
(b) Acids.Reagent grade HNO
3
, HCl, and H
2
SO
4
. Test acids
for freedom from Zn by AA measurement of appropriately diluted
sample. If contaminated, purify HNO
3
and HCl by distillation. Fur-
ther test purity of reagents and efficiency of cleaning by conducting
blank determinations by appropriate ashing method.
C. Preparation of Test Solution
Prepare representative test sample by mixing, blending, or grind-
ing.
(a) Wet ashing.Accurately weigh, into 300 or 500 mL
Kjeldahl flask, test portion 10 g, estimated to contain 25100g
Zn. (If material is liquid, evaporate to small volume.) Add ca
5 mL HNO
3
and cautiously heat until first vigorous reaction sub-
sides. Add 2.0 mL H
2
SO
4
and continue heating, maintaining oxi-
dizing conditions by adding HNO
3
in small increments (large
amounts may introduce Zn) until solution is colorless. Continue
heating until dense fumes of H
2
SO
4
are evolved and all HNO
3
has
been removed. Cool, dilute with ca 20 mL H
2
O, filter through fast
paper (prewashed) into 100 mL volumetric flask, and dilute to
volume with H
2
O. Dilute further, if necessary, with H
2
SO
4
(1 +
49) to attain working range of spectrophotometer.
(b) Dry ashing.Accurately weigh, into clean Pt dish, representa-
tivetest portionestimatedtocontain25100gZn. Char under IRlamp
and ash at temperature 525Cuntil C-free. (Raise temperature of fur-
nace slowlyto525Ctoavoidignition.) Dissolve ashunder watchglass
in minimumvolume HCl (1 + 1). Add ca 20 mL H
2
Oand evaporate to
near dryness on steambath. Add 20 mL 0.1M HCl and continue heat-
ing ca 5 min. Filter through fast paper into 100 mL volumetric flask.
Wash dish and filter with several 510 mLportions of 0.1MHCl, cool,
and dilute to volume with 0.1M HCl. Dilute further, if necessary, with
0.1M HCl to attain working range of instrument.
D. Determination
Set instrument to previously established optimum conditions or
according to manufacturers instructions. Determine Aof ashed so-
lution or dilution, and 5 standards within optimum working
range, taking 2 readings (before and after sample readings). Flush
burner with H
2
O and check 0 point between readings. Determine
Zn content from standard curve obtained by plotting A against g
Zn/mL:
Concentration Zn (mg / kg)
gZn/mL fr
=
om curve dilution factor, mL)
g test portion
(
Reference: JAOAC 51, 1042(1968).
CAS-7440-66-6 (zinc)
2000 AOAC INTERNATIONAL