9.2.

22
AOAC Official Method 971.21
Mercury in Food
Flameless Atomic Absorption Spectrophotometric Method
First Action 1971
Final Action 1976
(Rinse all glassware before use with HNO
3
[1 + 9].)
A. Apparatus
(a) Atomic absorption spectrophotometer.(Instrumentation
Laboratory, Inc., 113 Hartwell Ave, Lexington, MA 02173, USA,
Model 153 [or successors], or equivalent.) Equipped with Hg hollow
cathode lamp and gas flow-through cell (Figure 971.21), 25 (id)
115 mm with quartz windows cemented in place. Operating condi-
tions: wavelength 253.7 nm, slit width 160 m, lamp current 3 ma,
and sensitivity scale 2.5.
(b) Diaphragm pump.(Neptune Dyna-Pump, or equivalent.)
Coat diaphragmand internal parts of pump with acrylic-type plastic
spray. Use 16 gage Teflon tubing for all connections.
(c) Water condenser.1218 (id) 400 mm borosilicate, 24/40
standard taper joint, modified to hold 6 mm Raschig rings. Fill con-
denser with Raschig rings to height of 100 mm; then place 20 mm
layer of 4 mm diameter glass beads on top of rings.
(d) Gas inlet adapter.24/40 standard taper, e.g., Kontes Glass
Co. No. 181000.
(e) Digestion flask.250 mL boiling flask with 24/40 standard
taper joint.
B. Reagents
(a) Reducing solution.Mix 50 mLH
2
SO
4
with ca 300 mLH
2
O.
Cool to room temperature and dissolve 15 g NaCl, 15 g
hydroxylamine sulfate, and25gSnCl
2
insolution. Dilute to 500 mL.
(b) Diluting solution.To 1 L volumetric flask containing
300500 mL H
2
O, add 58 mL HNO
3
and 67 mL H
2
SO
4
. Dilute to
volume with H
2
O.
(c) Magnesium perchlorate.Drying agent placed in filter flask
(Figure 971.21). Replace as needed. [Caution: Mg(ClO
4
)
2
is explo-
sive when in contact with organic substances.]
( d) Mer cur y s t andar d s ol ut i ons . ( 1) St ock s ol u-
tion.1000 g/mL. Dissolve 0.1354 g HgCl
2
in 100.0 mL H
2
O.
(2) Working solution.1 g/mL. Dilute 1 mL stock solution to 1 L
with 0.5M H
2
SO
4
. Prepare fresh daily.
C. Determination
Weigh 5.0 g test portion into digestion flask; add 25 mL 9M
H
2
SO
4
, 20 mL7MHNO
3
, 1 mL2%sodiummolybdate solution, and
56 boiling chips. Connect condenser (with H
2
Ocirculating through
it) and apply gentle heat ca 1 h. Remove heat and let stand 15 min.
Add 20 mL HNO
3
HClO
4
(1 +1) through condenser. Turn off H
2
O
circulating through condenser and boil vigorously until white fumes
appear in flask. Continue heating 10 min.
Cool. Cautiously add 10 mL H
2
O through condenser while swirl-
ing liquid in flask. Again boil solution 10 min. Remove heat and
wash condenser with three 15 mL portions H
2
O.
Cool solution to room temperature. Completely transfer digested
sample with H
2
Oto 100 mLvolumetric flask and dilute to volume with
H
2
O. Transfer 25.0 mL aliquot fromeach test portion to another diges-
tion flask. Adjust volume to ca 100 mL with diluting solution, B(b).
Adjust output of pump to ca 2 L air/min by regulating speed of
pump with variable transformer. Connect apparatus as in Fig-
ure 971.21, except for gas inlet adapter. With pump working and
spectrophotometer zeroed, add 20 mL reducing solution to diluted
aliquot. Immediatelyconnect gas inlet adapter andaerate ca 3min. (Ad-
just aerationtimetoobtainmaximumA.) RecordA, disconnect pressure
on out side of pump, and open vent on filter flask to flush system.
Prepare reagent blank and standard curve by adding 0, 0.2, 0.4,
0.6, 0.8, and 1.0 g Hg to series of digestion flasks. To each flask add
100 mL diluting solution. Finally, add reducing solution and aerate
standards as for test portion.
Plot standard curve from least squares linear regression of A
against g Hg. (See Definitions of Terms and Explanatory Notes,
item [25], or use calculator which performs linear regression.) De-
2000 AOAC INTERNATIONAL
Figure 971.21Apparatus for flameless atomic absorption analysis.
termine g Hg in aliquot from curve. If g Hg determined falls out-
side range of calibration, repeat determination with smaller aliquot
of test solution to bring g Hg into region of standard curve. From
size of aliquot used, determine total g Hg in original test portion.
Concentration Hg (g/kg) =
g Hg
g test portion
Reference: JAOAC 54, 202(1971).
CAS-7439-97-6 (mercury)
2000 AOAC INTERNATIONAL