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9.1.

02
AOAC Official Method 982.23
Cadmium and Lead in Food
Anodic Stripping Voltammetric Method
First Action 1982
Final Action 1988
(Not applicable to fats and oils.)
A. Principle
Material is dry-ashed with K
2
SO
4
and HNO
3
at ca 500C. Pb and
Cd are determined by anodic stripping voltammetry (ASV). Esti-
mated quantitation limits, based on 10 g sample, are 0.005 mg Cd/kg
and 0.010 mg Pb/kg.
B. Apparatus
[Thoroughly soak all glassware and plasticware in 20% (v/v)
HNO
3
for 24 h and rinse with distilled, deionized H
2
O.]
(a) Voltammetric analyzer.Capable of ASVand equipped with
necessary accessories, i.e., cells, electrodes, recorders, Hg capillar-
ies, micrometer or similar device for adjustingdropsize, stirring mo-
tor, etc. (EG&G Princeton Applied Research Corp., PO Box 2565,
Princeton, NJ 08540, Models 384A, 264A, and 303A, or equivalent,
for differential pulse anodic stripping voltammetry [DPASV] at
hanging Hg drop electrode; Environmental Sciences Associates,
45 Wiggins Ave, Bedford, MA01730, Model 3010A, or equivalent,
for linear sweep anodic stripping voltammetry [LSASV]).
(b) Ashing vessels.150250 mL quartz, Vycor, or Pyrex beak-
ers equipped with suitable glass covers (Fisher Scientific Co.,
No. 2-609A, or equivalent). Quartz is preferred. Vycor or Pyrex may
be used if quartz beakers are not available.(Note: For best results,
quartz beakers should be fire-polished to retard etching.)
(c) Drying oven.Controllable within range 50150C with
5C variation.
(d) Furnace.Controllable within range of 1001000C with
5Cvariation. Check calibration of oven temperature control to en-
sure accurate temperatures. Furnace must be operated in suitable
fume hood.
(e) Controllable hot plate.
(f) Micropipets.10through100L(Eppendorf, or equivalent).
C. Reagents
(Note: Use only distilled, deionized H
2
O.)
(a) Nitric acid.J.T. Baker Chemical Co. No. 9598, or equiva-
lent.
(b) Potassium sulfate ashing solution.10 g/100 mL. Dissolve
50.0 g K
2
SO
4
(J.T. Baker Chemical Co. No. 3278, or equivalent) in
400mLH
2
Ocontaining10mLHNO
3
. Dilute to500mLwith H
2
O.
(c) Nitrogen.Prepurified, H
2
O-pumped.
(d) Electrolyte solution.1.7Min CH
3
COOH, 1.25Min sodium
acetate trihydrate, and 0.1M in tartaric acid. Dissolve 170.0 g
NaCH
3
COO3H
2
O (ACS) in 300 mL H
2
O. Add 97 mL glacial
CH
3
COOHand 15 g tartaric acid (ACS). Dilute to 1 L with H
2
O. pH
should be 4.7 0.1.
(e) Cadmium standard solution.1.0 mg/mL. Dissolve 1.000 g
Cd (99.99%) in 10 mL HNO
3
in 1 L volumetric flask. Dilute to vol-
ume with H
2
O.
(f) Lead standard solution.1.0 mg/mL. Dissolve 1.000 g Pb
(99.99%) in 10 mL HNO
3
in 1 L volumetric flask. Dilute to volume
with H
2
O.
(g) Working standard solutions.Prepare either separate or
mixed working standard solution for Cd and Pb in the range
0.110 g/mL from standard solutions (e) and (f) by dissolving ap-
propriate aliquots in 1% (v/v) HNO
3
.
[Note: Electrolyte solution, (d), and K
2
SO
4
solution, (b), may re-
quire further cleanup for sufficiently low reagent blanks. For stated
quantitation limits, analyte concentrations in final cell solutions
(electrolyte and sample solutions) of reagent blank should not be
>0.5 ng Cd/mLand >1 ng Pb/mL. Controlled potential electrolysis is
recommended means of cleaning reagents.
D. Preparation of Material
(Note: Laboratory contamination control is important. Take all
precautions possible to avoid contamination of laboratory and test
samples, reagents, and equipment. Prepare at least 3 control reagent
blanks which include any additional H
2
O and HNO
3
used for test
portion ashing. Carry control reagent blanks through entire
method.)
Weigh 5.010.0 g homogenized test portion into ashing vessel,
B(b). Use 5.0 g for dry materials such as cereals. Add 5.0 mLK
2
SO
4
ashing solution, C(b), and mix thoroughly, using glass stirring rod.
If needed, add H
2
O to ensure sample and ash aid are well mixed.
Cover ashing vessel with glass cover and dry in 110120C oven,
B(c), until thoroughly dry (usually 23 h or, if desired, overnight).
Place vessel in cold furnace, B(d), and set temperature at
500550C. (Caution: Do not heat >500C if using Pyrex beakers,
and avoid excessive overshooting of temperature.) Maintain set
temperature 4 h (may be ashed overnight). Remove vessel from
furnace, and cool. Ash should be white and essentially carbon-free.
Wash down sides of vessel with minimum amount H
2
O and add
2.0 mL HNO
3
. Use glass stirring rod to break up solid particles. Dry
thoroughly on hot plate, B(e), at lowsetting. If samples such as sug-
ars and cereals splatter on hot plate during HNO
3
treatment, dry un-
der IRlamp instead. Increase hot plate setting to mediumfor several
minutes to ensure dryness. Return vessel to 500C furnace 30 min.
Cool; if necessary, repeat HNO
3
treatment using 1 mLincrements of
HNO
3
, until white, C-free ash is obtained.
Add 1.0 mL HNO
3
and ca 10 mL H
2
Oto vessel, swirl to dissolve,
and let stand 5 min. If residue remains undissolved, warm gently on
8090C hot plate not >5 min. Minimize possible Sn(II) formation
by heating dilute acid solution as little as possible. Small amount of
white, siliceous-like precipitate may remain undissolved. Cool, and
quantitatively transfer sample to 50 mL volumetric flask with aid of
H
2
O. Dilute to volume with H
2
O and mix well. Let stand to allow
any precipitate present to settle. Do not filter. Use clear supernate to
determine analytes by either DPASV or LSASV below.
E. Differential Pulse Anodic Stripping Voltammetry
Transfer 5.0 mL aliquot of test solution to electrolysis cell con-
taining Teflon-coated stirring bar and add 5.0 mL electrolyte solu-
tion, C(d), to cell. (Aliquot volume may be varied as long as 1:1 ratio
is maintained between sample solution and electrolyte.) pH of cell
solution should be 4.3 0.3. Room temperature should be constant
(1C/2 h) and between 20 and 30C. Purge solution 5 min with
N
2
, C(c). Adjust gas inlet to let N
2
flowgently above and across solu-
tion surface. If hanging Hg drop electrode is used, add fresh drop of
Hg to capillary tip with micrometer or similar device to ensure
reproducibility of drop. Turn on stirrer motor and electrolyze solu-
tion at 0.8 V vs saturated calomel electrode (SCE) or Ag/AgCl
electrode. Deposition time may vary with instrument (see manufac-
2000 AOAC INTERNATIONAL
turers instructions). When using PAR 174 polarographic analyzer,
12 min is sufficient, depending on level of analytes of interest in
cell solution. Stop stirring and let solution equilibrate 30 s. Linerarly
increase applied voltage anodically. Followmanufacturers instruc-
tions for rate of scan, e.g., 26 mV/s. Measure wave height at peak
potentials for Cd at 0.62 0.05 V and for Pb at 0.45 0.05 V vs
SCE or Ag/AgCl. For widely varying concentrations of Cd and Pb,
change current sensitivity to appropriate range by momentarily stop-
ping stripping scan at end of Cd peak, switching to appropriate sensi-
tivitysettingfor Pb, andthencontinuingscanbefore Pbpeak begins.
Quantitate total amounts of Pb and Cd in cell solution by using
method of standard additions in cell as follows: Record
voltammogram from known volume of cell solution. From working
standard solution, C(g), add known amounts of Pb and Cd, using ap-
propriate micropipets, B(f) and being certain to add amount of each
element sufficient to generate peak heights ca twice those given by
sample cell solution. Repeat with 2 more similar additions of work-
ing standard solution to cell solution. For each analyte, plot g added
on x-axis vs peak height in A current on y-axis. Extrapolate linear
plot to x-axis intercept to determine total amount of analyte in sam-
ple aliquot. If available, use computer program based on method of
least squares to calculate regression line and determine amount of
analyte in sample aliquot. Similarly, determine amount of each
analyte in reagent blank aliquots, using same volume of aliquots for
reagent blank as for sample.
Calculate concentrationof analyte (mg/kg) insample as follows:
Concentration (mg/kg) =
B C
A
50
W

where A = mL test solution taken for analysis; B = g analyte in test


solution aliquot; C = average g analyte in reagent blank solution
aliquots; and W = total g test portion.
F. Determination by Linear Sweep Anodic Stripping
Voltammetry
Transfer 2.0 mL aliquot of sample solution to electrolysis cell and
add 3.0 mLelectrolyte, C(d). pHof cell solution should be 4.3 0.3. De-
posit elements of interest ontocomposite Hggraphite electrode (CMGE)
at 0.9 Vvs Ag/AgCl reference electrode for 30 min. Bubble N
2
through
cell solution during entire deposition period. Linearly increase applied
voltage anodicallyat 60mV/s from0.9to0.2Vvs Ag/AgCl reference
electrode. Measure peak current (A) for each analyte.
Run reagent blank in same manner using same size aliquot as for
sample and determine peak current (A) for each analyte. For each
analyte, make standard addition to cell solution and measure peak
current (A). Calculate conversion factor, g/A, for each analyte
asg of addition divided by difference between peak current before
and after addition of analyte standard. Verify conversion factors pe-
riodically. Multiply sample peak current (A) by conversion factor
to determineg of each analyte in sample solution aliquot. Calculate
ppm (g/g), using equation in E.
G. Interference
Tl may interfere with Pb determination, but its occurrence in food
is unlikely. If Tl interference is suspected, treat as follows: Transfer
5.0 mLaliquot of sample solution to electrolysis cell and make basic
with 3.0 mL NaOH. Determine elements of interest in this solution
by ASV in the usual manner. Plating potential is 1.0 V vs SCE or
similar reference electrode. Strip deposited elements by anodically
scanning from 1.0 to 0.3 V vs SCE. In this manner, Cd and Pb
peaks shift to 0.78 0.05 V and 0.73 0.05 V vs SCE, respec-
tively. Tl peak remains at 0.47 V vs SCE.
References: JAOAC 65, 970, 978(1982); 70, 295(1987).
CAS-7440-43-9 (cadmium)
CAS-7439-92-1 (lead)
2000 AOAC INTERNATIONAL